ʻO ka loaʻa ʻana a me ka hāʻawi ʻana i nā meaʻai e hoʻohui i ka ʻai ʻana i nā pepeke a me nā ʻano mōʻaukala ola.ʻO ka hoʻonāukiuki arabiensis ma ka mimi pipi no ka loaʻa ʻana o ka meaʻai e hoʻomaikaʻi i nā ʻano o ka mōʻaukala ola.
E hōʻoia i ka palekana. Ua huki ʻia ʻo Arabiensis i ke ʻala o ka mimi bipi hou, 24-hola, 72-hola, a me ka ʻimi hoʻokipa a hānai ʻia i ke koko (48-hola ma hope o ke koko) ua ana ʻia nā wahine i loko o ka Y-tube olfactometer, a ua helu ʻia nā wahine hāpai i ka hoʻāʻo ʻana i ka cophysiological bioactive i ka hoʻāʻo ʻana i ka cophysiological. ʻEhā mau makahiki. Ua loiloi ʻia nā hui pū ʻana o nā pūhui bioactive i ka Y-tube a me nā hoʻokolohua kahua. hoʻoiho.
E ʻimi i ke koko o ka mea hoʻokipa a me ka meaʻai. Ma ka hale hana a me nā haʻawina kula, ua huki ʻia ka poʻe ʻArabia i ka ʻala maoli a me ka synthetic o ka mimi pipi hou a ʻelemakule.
Anopheles arabinis ka loaʻa ʻana a me ka hāʻawi ʻana i ka mimi bipi no ka hoʻomaikaʻi ʻana i nā hiʻohiʻona o ke ola.
ʻO ka loaʻaʻana a me ka hāʻawiʻana i nā meaʻai e hoʻohui i kaʻaiʻana i nā'ōpala a me nāʻano moʻolelo o ke ola [1,2,3]. Hiki i nā'ōpala ke koho a loaʻa i ka meaʻai a hana i ka hānaiʻana e pili ana i ka loaʻaʻana o ka meaʻai a me nā mea e pono ai ka meaʻai [1, 3]. ʻO ka hāʻawiʻana i nā meaʻai e pili ana i ke kaʻina o ka moʻolelo ola a hiki ke alakaʻi i nā koi likeʻole no ka maikaʻi a me ka nui o ka meaʻai i nā manawa likeʻole o ke ola. Hiki ke loaʻa kēia mau meaʻai ma o ka hānai ʻana, e like me ka lepo, nā ʻōpala like ʻole a me nā mea huna o nā vertebrates, a me ka carrion, kahi hana i kapa ʻia he puddles [2]. squito Anopheles gambiae sensu lato (sl) puka mai ma ke 'ano he 'malnourished' kanaka makua [8], no laila, he mea nui paha ka wai i kona mau hiʻohiʻona moʻolelo o ke ola, akā ua mālama ʻole ʻia kēia ʻano.
Ua kaupalena ʻia ka ʻai ʻana o ka nitrogen i nā namu Anopheles wahine makua ma muli o ka haʻahaʻa o ka caloric i lawe ʻia mai ka pae larval a me ka hoʻohana pono ʻole ʻana i ka ʻai koko [9]. hiki ke hoʻohana i ka hānai hou ʻana o ka vertebrate excreta e loaʻa ai nā pūhui nitrogenous e hoʻonui ai i ka adaptation a me ka holo manuverability, e like me ka mea i hōʻike ʻia e nā pepeke ʻē aʻe. ʻO ka mimi pipi he kumu waiwai i nā pūhui nitrogenous, me ka urea e helu ana no 50-95% o ka nui o ka nitrogen i loko o ka mimi hou [15, 16]. I ka makahiki o ka mimi pipi, hoʻohana nā microorganisms i kēia mau kumuwaiwai e hōʻemi i ka paʻakikī o nā pūhui nitrogenous i loko o 24 mau hola [15]. squitoes) holomua [15], ʻo ia paha he wahine Ann.arabiensis i makemake nui ʻia i ka mimi 24 mau hola a i ʻole [13, 14].
I loko o kēia noiʻi ʻana, ʻimi ʻia nā Ans i hānai ʻia i ke koko. I ka wā o kāna pōʻai gonadotropin mua, ua loiloi ʻia ʻo Arabiensis no ka loaʻa ʻana o nā pūhui nitrogenous, me ka urea, ma o ka hui ʻana o ka mimi. Hāʻawi kēia i nā hōʻailona hilinaʻi no ka mea hoʻokipa a me ka mea hānai koko ʻo An. I kā lākou ʻimi ʻana i kēia kumuwaiwai meaʻai kūpono, ua ʻike ʻo Arabiensis i nā correlations kemika ma hope o ka nani ʻokoʻa i ʻike ʻia.ʻO ka namu huki.Loaʻa a puʻunaue arabiensis i nā pūhui nitrogenous i loaʻa i loko o ka mimi vertebrate e hoʻoikaika i nā hiʻohiʻona moʻolelo o ke ola. Kūkākūkā ʻia kēia mau hopena i ka pōʻaiapili o nā hopena maʻi maʻi a pehea e hiki ai ke hoʻohana ʻia no ka nānā ʻana a me ka mālama ʻana.
Ua mālama ʻia nā Anopheles arabicans (Dongola strain) ma 25 ± 2 °C, 65 ± 5% RH a me kahi 12:12 h māmā: pōʻeleʻele pōʻeleʻele. Ua hānai ʻia nā larvae i loko o nā pā plastik (20 cm × 18 cm × 7 cm) i hoʻopiha ʻia me ka wai hoʻoheheʻe ʻia a hānai ʻia ʻo Tetramin® i ka meaʻai iʻa (Tetramin® iʻa i ʻohi ʻia (Tetramin® kīʻaha i ʻohi ʻia, ʻAʻole ʻai iʻa ʻo Tetramin® (Tetramin®), ʻAʻole ʻai iʻa ʻo Melato (Tetramin®), ʻAʻole ʻo Melato (Tetramin®) iʻa (Tetramin® iʻa (Tetramin®), ʻAʻole ʻai iʻa iʻa (Tetramin®). rtila, Åstorp, SE) a laila hoʻoneʻe ʻia i nā hīnaʻi Bugdorm (30 cm × 30 cm × 30 cm; MegaView Science, Taichung, Taiwan) e ʻae ai i ka puka ʻana mai o nā mākua. No ka hoʻomākaukau ʻana i nā makika koko koko no nā bioassays ma hope, ua hoʻolako ʻia nā wahine 4 dpe me ke koko hipa defibrotic (Håtunalab, Bro, SE) me ka hoʻohana ʻana i kahi ʻōnaehana hānai membrane (Hemotek Discovery Workshops, Accrington, UK a i ʻole ka hoʻololi ʻana i ka wahine. % sucrose ad libitum no 3 mau lā ma mua o nā hoʻokolohua i wehewehe ʻia ma lalo nei. Ua hoʻohana ʻia nā wahine hope no nā bioassays paipu lele a hoʻoneʻe ʻia i ke keʻena hoʻokolohua, a laila ua hoʻomaʻemaʻe i ka wai ad libitum no 4-6 mau hola ma mua o ka hoʻokolohua.
Ua hoʻohana ʻia nā hoʻāʻo hānai e hoʻonui i ka mimi a me ka ʻai ʻana i ka ʻai ʻana i ka wahine An.Arab makua. Ua hāʻawi ʻia nā wahine ʻimi hoʻokipa a hānai i ke koko i kahi meaʻai i loaʻa i ka 1% diluted hou a me ka ʻelemakule pipi mimi, nā ʻano like ʻole o ka urea, a me ʻelua mau mana (10% sucrose a me ka wai) no 48 h. waiwai, Stockholm, SE) ua hoʻohui ʻia i ka meaʻai a hāʻawi ʻia i loko o kahi matrix 4 × 4 i 250 µl microcentrifuge tubes (Axygen Scientific, Union City, CA, US; Kiʻi 1A) E hoʻopiha i ka ʻaoʻao (~ 300 µl). kiʻekiʻe; Semadeni, Ostermundigen, CH; Kiʻi 1A) i loko o ka pōʻeleʻele piha ma 25 ± 2 cm °C a me 65 ± 5% haʻahaʻa pili.
E nānā i ka mimi pipi a me ka urea i hoʻomoʻa ʻia e ka mea hoʻokipa a me ka wahine omo koko Anopheles arabianus. I ka hoʻāʻo hānai ʻana (A), ua hāʻawi ʻia nā makika wahine me kahi meaʻai e komo ana i ka mimi bipi hou a ʻelemakule, nā ʻano ʻokoʻa o ka urea, sucrose (10%), a me ka wai distilled (H2O). ed 72-hola pipi mimi emi iho malalo o 168-hola pipi mimi (B). ʻO ka mean huina nitrogen maʻiʻo (± standard deviation) o ka mimi ua hoikeia ma ka inset. host-seeking (D, F) a me ke koko-momi (E, G) lawe i ka urea ma ka dose-dependent manner. Mean inhaled volumes inhaled volume (D, E) with significantly different letters from AN-Way's names 0.05). Hōʻike nā pahu hewa i ka hewa maʻamau o ka mean (BE).
No ka hoʻokuʻu ʻana i ka meaʻai i hoʻopaʻa ʻia, ua hoʻokomo ʻia nā makika i loko o 1.5 ml microcentrifuge tubes i loaʻa ka 230 µl o ka wai i hoʻoheheʻe ʻia a ua hoʻopau ʻia ka ʻiʻo me ka hoʻohana ʻana i ka pestle a me ka mīkini cordless (VWR International, Lund, SE), a ukali ʻia e ka centrifugation ma 10 krpm no 10 min . -Aldrich) a me ka absorbance (λ620) i hoʻoholo 'ia me ka spectrophotometer-based microplate heluhelu (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm). ʻO keʻano'ē aʻe, ua hoʻopiliʻia nā makika i ka 1 ml o ka wai i hoʻomaʻemaʻeʻia, 900 µl o ia mea i hoʻololiʻia i kahi cuvette (Nm λ 600metric analysis). No ka helu ʻana i ka ʻai meaʻai, ua hoʻomākaukau ʻia kahi pihi maʻamau e ka dilution serial e hāʻawi i ka 0.2 µl a i ka 2.4 µl o 1 mg ml-1 xylene cyanide. A laila, ua hoʻohana ʻia ka mānoanoa o nā mea hoʻonaninani i ʻike ʻia no ka hoʻoholo ʻana i ka nui o ka meaʻai i ʻai ʻia i kēlā me kēia makika.
Ua kālailai ʻia ka ʻikepili volume me ka hoʻohana ʻana i ka hoʻohālikelike ʻana o ka ʻano like ʻole (ANOVA) a ukali ʻia e Tukey's post hoc pairwise hoʻohālikelike (JMP Pro, v14.0.0, SAS Institute Inc., Cary, NC, US, 1989-2007). , Pūnaehana GraphPad, San Diego, CA, US).
Ma kahi o 20 µl o nā laʻana mimi mai kēlā me kēia hui makahiki i hoʻopaʻa ʻia ma Chromosorb® W / AW (10 mg 80/100 mesh, Sigma Aldrich) a hoʻopili ʻia i loko o nā capsules tin (8 mm × 5 mm). a me ka mimi kahiko e like me ka protocol a ka mea hana.
No ka loiloi i ka hopena o ka ʻai ʻana i ka wahine ʻimi a me ke koko koko, ua hoʻokomo ʻia nā makika i loko o nā kīʻaha Petri nui (12 knm ke anawaena a me 6 knm ke kiʻekiʻe; Semadeni) me kahi puka i uhi ʻia i loko o ke poʻi (3 knm ke anawaena) me No ka ea a me ka lako meaʻai. sucrose a me ka wai. Ua paipu ia kela ai keia ai ma luna o ka tampon niho (DAB Dental AB, Upplands Väsby, SE) i hookomoia i loko o ka 5 ml syringe (Thermo Fisher Scientific, Gothenburg, SE), ka plunger wehe, a kau ma luna o ka petri dish (helu 1).1A). ua hoʻolei ʻia nā makika a hiki i ka make ʻana o ka makika hope loa (n = 40 no ka mālama ʻana).
He wili namu lele maʻamau e pili ana iā Attisano et al.[17], i hana ʻia me 5 mm mānoanoa o nā panela acrylic akaka (10 knm ākea x 10 knm ka lōʻihi x 10 knm kiʻekiʻe) me ka ʻole o ka mua a me hope paneli (Fig. 3: luna). neodymium magnets 9 knm kaawale. Ua hoʻokaʻawale ʻia ka paipu kuʻekuʻe i hana ʻia me ka mea like (6.5 knm L) e hana i ka lima hoʻopaʻa ʻia a me ka lima e lawe ana i kahi ʻāpana ʻāpana alumini ma ke ʻano he hōʻailona keakea māmā.
Hāʻawi ʻia nā wahine pōloli 24-hola i ka ʻai ma luna no 30 mau minuke ma mua o ka hoʻopaʻa ʻana. -built data logger, a laila mālama ʻia a hōʻike ʻia me ka hoʻohana ʻana i ka polokalamu PC-Lab 2000™ (v4.01; Velleman, Gavere, BE).
No ka nānā ʻana i ke ʻano o ka hana lele, ua helu ʻia ka huina o ka mamao i lele ʻia (m) a me ka huina o nā hana lele holoʻokoʻa i kēlā me kēia hola ma kahi manawa 24-hola. helu ʻia nā pōʻai ma nā hoʻonui 10 mau minuke.
No ka loiloi i ka hopena o ka ʻai i ka hana hānau o An.arabiensis, ua hoʻoneʻe pololei ʻia nā wahine ʻeono (4 dpe) i nā hīnaʻi Bugdorm (30 cm × 30 cm × 30 cm) ma hope o ka hōʻiliʻili ʻana i ke koko a laila hāʻawi i ka ʻai hoʻokolohua no 48 mau hola e like me ka mea i hōʻike ʻia ma luna. 4 hola. E hana hou i kēlā me kēia meaʻai 20-50 mau manawa. Ua helu ʻia nā hua manu a hoʻopaʻa ʻia no kēlā me kēia hale hoʻokolohua.Leica Microsystems Ltd., DE).Ua mālama 'ia nā hua i koe i loko o ka lumi ho'omalu i ke aniau ma lalo o nā kūlana hanai ma'amau no 24 h, a ua ana 'ia kekahi subsample o ka 1st instar larvae i puka hou mai nei (n ≥ 200 no ka mea'ai), e like me ka mea i ho'ākāka 'ia ma luna. Ua ho'ohālikelike 'ia ka helu o nā hua a me ka nui o nā hua a me nā larvae, ma waena o nā lā'au lapa'au a me ka ho'ohana 'ana i ka ho'oma'ama'a 'ana a me ka ho'ohana 'ana i ka one-ways ho'opa'a a me ka ho'ohana 'ana i ka one-way's ho'oJV. SAS Institute Inc.).
Headspace volatiles mai ka hou (1 hr post-sampling), 24 hr, 72 hr a me 168 hr makahiki mimi ua ohi ia mai laana mai i ohiia mai Zebu pipi, Arsi races. No ka oluolu, mimi laana ua ohi i kakahiaka nui oiai na bipi i loko o ka halepapaa. sco, Frischhalteprodukte GmbH a me Co., Minden, DE) ma 3 l polyamide me ka poʻi I loko o ka vinyl chloride plastic drums.Headspace volatiles mai kela bovine mimi hāpana i ohi pono (hou) a ma hope o maturation ma ka lumi wela no 24 h, 72 h a me 168 h, ie kela mimi hāpana mea o kēlā me kēia makahiki hui.
No ka hōʻiliʻili headspace volatiles, ua hoʻohana ʻia kahi ʻōnaehana hoʻopaʻa ʻia e hoʻoheheʻe i kahi kahe kinoea kānana carbon-filtered (100 ml min-1) ma o ka ʻeke polyamide i ke kolamu adsorption no 2.5 h ma o ka hoʻohana ʻana i ka paila diaphragm vacuum (KNF Neuberger, Freiburg, DE). Ma ke ʻano he mana, ua hana ʻia ke kolamu headspace o ka xsorpamide cm mai kahi pahu pahu x5flon. 3 mm id) e loaʻa ana i ka 35 mg o Porapak Q (50/80 mesh; Waters Associates, Milford, MA, US) ma waena o nā pulupulu aniani. o pentane. Hoʻohui ʻia nā hōʻiliʻili poʻo a laila mālama ʻia ma -20°C a hiki i ka hoʻohana ʻia no ka nānā hou ʻana.
Nā pane Behavioral o ka 'imi a me ka 'ai koko An. Headspace volatile extracts ohiia mai hou, 24-h, 72-h, a me 168-h-aged mimi ua kālailai 'ia no ka volatile extracts mai Arabidopsis makika me ka hoʻohana 'ana i ka pololei aniani paipu olfactometer [18]. Ua alakaʻiʻia nā hoʻokolohua i ka manawa o ZT'9rabk-15. olfactometer paipu (80 knm × 9.5 knm id) ua ho'omālamalama 'ia me 3 ± 1 lx o ke kukui 'ula'ula mai luna mai. Ua ho'ohehe'e 'ia ka lanahu a me ka ho'ohehe'e 'ia (25 ± 2 °C, 65 ± 2% ka ha'aha'a ha'aha'a) i hala i ka bioassay ma 30 knm s-1. Ho'okumu 'ia ka ea ma waena o ka pale kila me ka aspennar. (4 knm × 1 knm; L: D; DAB Dental AB), ua hoʻokuʻu ʻia mai kahi wili 5 cm ma ka hopena makani o ka olfactometer, me nā hoʻololi stimulator i kēlā me kēia 5 minuke. 3 mau hola ma mua o ka hoʻomaka ʻana o ka hoʻokolohua. Ua kau ʻia ka hale hoʻokuʻu ma ka ʻaoʻao downwind o ka olfactometer, a ua ʻae ʻia nā makika e hoʻomaʻamaʻa no 1 min, a laila ua wehe ʻia ka pahu lele o ka hale e hoʻokuʻu ai. o kekahi lā, ua ho'āʻoʻia ka helu like o nā lapaʻau a me nā mana i kēlā me kēia lā hoʻokolohua. E ʻimi i nā pane mai ka mea hoʻokipa a me ke koko i hānai ʻia ʻo Ans.Arabic versus headspace sets ua kālailai ʻia me ka hoʻohana ʻana i ka nominal logistic regression i ukali ʻia e nā hoʻohālikelike pairwise no nā ratio like ʻole (JMP Pro, v14.0.0, SAS Institute Inc.).
ʻO ka pane ʻana o An's spawning response.Headspace extracts from fresh and old cow mimi were analyzed in Bugdorm cages (30 cm × 30 cm × 30 cm; MegaView Science). ʻO nā kīʻaha plastik (30 mL; Nolato Hertila) i hoʻopiha ʻia me 20 mL o ka wai distilled i hāʻawi ʻia i ka spawning substrate a ua hoʻonohonoho ʻia i loko o ke kīʻaha o ka spawning substrate a ua hoʻonohonoho ʻia i loko o kahi ʻāpana o ke kīʻaha 2 cm. μl o kēlā me kēia ʻāpana poʻo ma kahi dilution 1:10. Ua hoʻohana ʻia ka nui like o ka pentane no ka hoʻoponopono ʻana i ke kīʻaha hoʻomalu. Ua hoʻololi ʻia nā kīʻaha lapaʻau a me nā kīʻaha hoʻomalu ma waena o kēlā me kēia hoʻokolohua e hoʻomalu ai i nā hopena kūlana. waiho ʻia i loko o ke kīʻaha hoʻomalu)/(ka huina o nā hua i waiho ʻia).
Ua hana ʻia ke kinoea chromatographic a me ka electron antenna pattern detection (GC-EAD) o ka wahine An.arabiensis e like me ka mea i wehewehe mua ʻia [20]. ʻO ka pōkole, ua hoʻokaʻawale ʻia nā extracts volatile hou headspace me ka hoʻohana ʻana i kahi Agilent Technologies 6890 GC (Santa Clara, CA, US) i hoʻolako ʻia me kahi kolamu HP-5 (30 m × 0.25 mm mānoanoa, ʻenehana id).a me ka elemakule mimi.Hydrogen ua hoʻohana 'ia e like me ka mobile pae me ka awelika linear kahe ana o 45 cm s-1.Each hāpana (2 μl) ua injected no 30 kekona ma splitless mode me ka inlet wela o 225 ° C. Ua papahana ka GC umu wela wela mai 35 °C (3 minuke paʻa) i ka 300 °C hoʻopaʻa minuke ma 100 mau minuke ma 100 °C. Hoʻohui ʻia ka 4 psi o ka nitrogen a hoʻokaʻawale i ka 1: 1 i loko o kahi Gerstel 3D/2 haʻahaʻa make leo keʻa (Gerstel, Mülheim, DE) ma waena o ka lapalapa ahi ionization detector a me ka EAD. Ua hala ka GC effluent capillary no EAD ma o kahi laina hoʻololi Gerstel ODP-2, e hahai ana i ka GC umu me 5 °C, i loko o kahi aniani i hui ʻia me 5 °C, i loko o kahi aniani i hui pū ʻia me 5 ° C, i loko o kahi pahu aniani i hui pū ʻia me 5 °C (mm1). 1.5 l min−1) Ua kau ʻia ka antenna ma kahi o 0.5 knm mai ka puka o ka paipu. Ua helu ʻia kēlā me kēia makika no hoʻokahi replicate, a no nā makika ʻimi hoʻokipa, ua hana ʻia ma ka liʻiliʻi he ʻekolu hoʻopiʻi ma nā ʻano mimi o kēlā me kēia makahiki.
ʻO ka ʻike ʻana i nā pūhui bioactive i loko o nā hōʻiliʻili poʻo o ka mimi pipi hou a ʻelemakule e hoʻohana ana i ka GC a me ka spectrometer mass (GC-MS; 6890 GC a me 5975 MS; Agilent Technologies) e kiʻi i nā pane antennal i ka GC-EAD analysis, e hana ana i ke ʻano electron impact ionization mode ma 70 eVUI. 0.25 mm anawaena o loko, 0.25 μm kiʻi mānoanoa) me ka helium e like me ka mobile phase me ka awelika linear kahe kahe o 35 cm s-1.A 2 μl hāpana i injected me ka hoʻohana 'ana i ka ia injector hoʻonohonoho a me ka umu wela e like me ka GC-EAD ana. i hoʻopaʻaʻia e ka hoʻokomoʻana i nā kūlana kūpono (Hoʻohui Kōnae 1: Papa S2) .
Ka loiloi i ka pono o ka hui 'ala synthetic i loko o nā pūhui bioactive i ʻike ʻia i loko o ka mimi hou a ʻelemakule e huki ai i ka poʻe ʻimi a me ke koko koko Ans.arabiensis, me ka hoʻohana ʻana i ka olfactometer a me ka protocol e like me ka mea i luna. 5D-G; Waihona hou 1: Papa S2). No ka nānā ʻana, e hoʻohana i ka 10 μl o kahi hoʻoheheʻe 1:100 o ka hui synthetic piha, me ka helu hoʻokuʻu holoʻokoʻa mai ka 140-2400 ng h-1, no ka loiloi ʻana i ka nani o ka mea hoʻokipa a me nā makika koko. Ua kālailai ʻia nā hui ʻana o Ans.Arab vs synthetic a subtractive me ka hoʻohana ʻana i ka nominal logistic regression i ukali ʻia e nā hoʻohālikelike pālua no nā ratio like ʻole (JMP Pro, v14.0.0, SAS Institute Inc.).
No ka loiloi inā hiki i ka mimi bipi ke lilo i kumu hoʻokipa no nā makika malaria, ka mimi bipi hou a ʻelemakule, i hōʻiliʻili ʻia e like me ka mea i hōʻike ʻia ma luna, a ua hoʻokomo ʻia ka wai i loko o nā bākeke 3 l (100 ml) a hoʻokomo ʻia i loko o nā pahele maunu.(BG-HDT version; BioGents, Regensburg, DE). He umi pahele i waihoia ma kahi o 50 m kaawale ma kahi e hanai ai, 400 m mai ke kaiāulu kauhale (Silay, Ethiopia, 5°53'24''N, 37°29'24'E) a ʻaʻohe pipi, ma nā wahi hānau mau a me nā kauhale. he ʻelima mau pō. Ua hoʻohālikelike ʻia nā helu makika i hopu ʻia i loko o nā pahele i maunu me ka mimi o nā makahiki like ʻole me ka hoʻohana ʻana i ka logistic regression me kahi puʻupuʻu beta binomial (JMP Pro, v14.0.0, SAS Institute Inc.).
Ma kahi kauhale malaria-endemic kokoke i ke kaona o Maki, Oromia māhele, ʻAitiopa (8° 11′ 08″ N, 38° 81′ 70″ E; Kiʻi 6A). ka haʻawina (Fig. 6A). ʻO nā pae hoʻohālike i hoʻohana ʻia e koho i nā hale: ʻaʻohe holoholona i ʻae ʻia i loko o ka hale, ʻaʻole ʻae ʻia ka kuke ʻana i loko (ka huki ʻana i ka wahie a i ʻole ka lanahu) (ma ka liʻiliʻi loa i ka wā hoʻokolokolo), a me nā hale me ka nui o ʻelua mau kānaka, e moe ana i ka unsecticides.ma lalo o ka makika i mālama ʻia. Ua hāʻawi ʻia ka ʻae kūpono e ka Institutional Research Ethics Review Board (IRB/022/2016) o ka Faculty of Natural Sciences (CNS-IRB), Addis Ababa University, e like me nā alakaʻi i hoʻokumu ʻia e ka World Medical Association Declaration of Helsinki. ('kebele') pae. ʻO ka hoʻolālā hoʻokolohua ma hope o kahi hoʻolālā 2 × 2 Latin square, kahi i hāʻawi ʻia ai nā huila synthetic a me nā mana i nā hale i hui pū ʻia i ka pō mua a hoʻololi ʻia ma waena o nā hale i ka pō hoʻokolohua aʻe. Ua hana hou ʻia kēia kaʻina hana he ʻumi mau manawa.
Ua hoʻoheheʻe ʻia kahi hui synthetic i loko o ka heptane (97.0% solvent GC grade, Sigma Aldrich) a hoʻokuʻu ʻia ma 140 ng h-1 me ka hoʻohana ʻana i ka pulupulu wick dispenser [20]. Ua ʻae ka dispenser wick i nā pūhui āpau e hoʻokuʻu ʻia i nā ʻāpana mau a puni ka 12 hola hoʻokolohua. Ua hoʻohana ʻia ʻo Heptane ma ke ʻano he mana o ka maʻi maʻi. pahele māmā (John W. Hock Company, Gainesville, FL, US; Kiʻi 6A). Ua kau ʻia nā pahele 0.8 - 1 m ma luna o ka honua, kokoke i ka wāwae o ka moena, a ua hiamoe kekahi mea manawaleʻa ma lalo o kahi ʻupena makika ʻole i mālama ʻia a hana ʻia ma waena o 18:00 a me 06:30. Ua nānā ʻia me ka hoʻohana ʻana i ka polymerase chain reaction (PCR) analysis no ka ʻike ʻana i ka ʻano morphologically i ʻike ʻia ʻo A. gambiae sl. Nā lālā o ka paʻakikī [23]. Ma ka hoʻopaʻa haʻawina, ua kālailai ʻia ka pahele ʻana o nā hale i hoʻopaʻa ʻia me ka hoʻohana ʻana i ke kumu hoʻohālike nominal logistic fit, kahi i hoʻohuli ʻia ai ka loli hilinaʻi a me ka lāʻau (synthetic mixture vs control) ka hopena paʻa (JMP.® 14.0. SAS Institute Inc.).Maʻaneʻi, hōʻike mākou i ka χ2 a me nā p-waiwai mai ka ho'āʻo ratio likelihood.
E noʻonoʻo inā he palekana. Ua hiki i ka Arabiensis ke loaʻa i ka mimi, kona kumu nui nitrogen, urea, ma ka hānai pololei ʻana, i loko o 48 mau hola o ka lawelawe ʻana no 4 mau lā ma hope (dpe) ʻimi hoʻokipa a me nā hoʻāʻo hānai wahine hānai koko (Fig. 1A). 1 a me F (5,299) = 56.00, p <0.0001, i kēlā me kēia; Fig. 1B, C). Eia kekahi, ʻai nā wahine ʻimi hoʻokipa i ka liʻiliʻi i ka mimi i nā hola 72 i hoʻohālikelike ʻia me ka mimi ma nā hola 168 (Fig. 1B). ʻAʻole hiki ke ʻike ʻia mai ka 10% sucrose (F(10,813) = 15.72, p <0.0001; Kiʻi 1D). He ʻokoʻa kēia i ka pane ʻana o nā wahine hānai koko, ka mea maʻamau e komo nui i nā meaʻai i loaʻa i ka urea ma mua o ka wai, ʻoiai ʻoi aku ka liʻiliʻi ma mua o 10 (10% sucrose) (5, 0.0000000000,000. 001; Kiʻi 1).1E). Eia kekahi, i ka hoʻohālikelike ʻana ma waena o nā kūlana physiological ʻelua, ʻoi aku ka nui o nā wahine phlebotomized i ka urea ma mua o nā wahine ʻimi hoʻokipa ma nā haʻahaʻa haʻahaʻa, a ua komo kēia mau wahine i ka nui o ka urea ma nā kiʻekiʻe kiʻekiʻe (F (1,953) = 78.80, p < 0.82, p <0.Fig. 1F, G). ʻOiai ʻo ka ʻai ʻana mai kahi meaʻai urea i ʻike ʻia he mau waiwai maikaʻi loa (Fig. 1D,E), ua hiki i nā wahine ma nā mokuʻāina ʻelua ke hoʻololi i ka nui o ka urea i komo ma luna o ka pae holoʻokoʻa o ka urea i loko o ke ʻano log-linear (Fig. 1F,G).).Pēlā, ʻike ʻia nā makika e hoʻomalu i kā lākou lawe ʻana i ka nitrogen ma o ka hoʻoponopono ʻana i ka nui o ka mimi i hoʻomoʻa ʻia, no ka mea, ʻike ʻia ka nui o ka nitrogen i loko o ka mimi i ka nui o ka lawe ʻia (Figure 1B, C a me B insets).
No ka nānā ʻana i nā hopena o ka mimi a me ka urea i ke ola ʻana o ka namu ʻimi a me ke koko koko, ua hānai ʻia nā wahine i ka mimi o nā makahiki ʻehā (hou, 24 h, 72 h, a me 168 h post-deposition) a me kahi ʻano o ka urea concentrations, a me ka wai distilled a me 10% sucrose i lawelawe ʻia ma ke ʻano he mana (ʻike i ka hopena o ke ola ʻana i ka wahine). : χ2 = 108.5, df = 5, p <0.0001; urea: χ2 = 122.8, df = 5, p <0.0001; Fig. 2B, C) a me nā wahine hānai koko (mimi: χ2 = <93.0, df = 93.0, p 1.0. , df = 5, p <0.0001; Kiʻi 2D, E). Ma nā hoʻokolohua a pau, ua hānai nā wāhine i kaʻai o ka mimi, ka urea, a me ka wai i haʻahaʻa loa i ke ola ma mua o nā wāhine i hānai i kaʻai sucrose (Figure 2B-E). (Fig. 2B) . Eia kekahi, ua ola lōʻihi nā wahine ʻimi hoʻokipa i hānai iā 135 mM urea ma mua o nā mana wai (p <0.04) (Fig.2C).Hoʻohālikelike ʻia me ka wai, ʻo nā wahine i hānai ʻia me ka mimi hou a me ka mimi 24-hola ua ola lōʻihi (p = 0.001 a me p = 0.012, i kēlā me kēia; Kiʻi 2D), ʻoiai ʻo nā wahine i hānai ʻia me ka mimi 72-hola ola ʻoi aku ka lōʻihi ma mua o ka poʻe i hānai ʻia. ʻO 135 mM urea, ua ola nā wahine hānai koko ma mua o nā mea ʻē aʻe o ka urea a me ka wai (p <0.013; Kiʻi 2E).
ʻO ke ola o ka mea hoʻokipa a me ka wahine omo koko Anopheles arabinis e hānai ana i ka mimi bipi a me ka urea. Ma ka bioassay (A), ua hāʻawi ʻia nā makika wahine i kahi meaʻai e komo ana i ka mimi bipi hou a ʻelemakule, nā ʻano ʻokoʻa o ka urea, sucrose (10%) a me ka wai distilled (H2O). d ma ka mimi (B, D) a me ka urea (C, E), a me na hoomalu, Sucrose a me ka wai, ua make.
ʻO ka lōʻihi a me ka helu o nā pōʻai i hoʻoholo ʻia ma ka hoʻāʻo ʻana i ka wili lele ma kahi 24-hola he ʻokoʻa ma waena o nā makika ʻimi hoʻokipa a me nā namu koko koko, i hōʻike ʻia ka liʻiliʻi o ka lele holoʻokoʻa holoʻokoʻa (Fig. 3). ʻO nā makika ʻimi hoʻokipa i hāʻawi i ka mimi hou a ʻelemakule a i ʻole sucrose a me ka wai i hōʻike i nā ʻano lele lele (Fig. 3), me nā wahine i ʻoi aku ka ʻoi aku o ka hana ʻana i ka 6 hola 4. ʻO nā makika i hānai i ka mimi ua hōʻike i nā ʻano lele like ʻole a ʻo ia ka mea maʻamau. 24 hola (Figure 3).
ʻO ka hana lele ʻana o ka wahine ʻimi koko ʻimi koko Anopheles arabinis e hānai ana i ka mimi pipi a me ka urea. I ka hoʻāʻo ʻana i ka wili lele, ua hoʻopaʻa ʻia nā makika wahine i ka mimi pipi hou a ʻelemakule, nā ʻano ʻokoʻa like ʻole o ka urea, sucrose (10%), a me ka wai distilled (H2O) ua hoʻopaʻa ʻia i ka hoʻopaʻa ʻana, ka lōʻihi o ke koko (nā lima a me ka wahine). helu ʻia o nā lele i kēlā me kēia hola no kēlā me kēia meaʻai ma kahi hola 24-hola i hoʻopaʻa ʻia (ʻeleʻele: hina; māmā: keʻokeʻo).
Ma keʻano laulā, ua hahai ka hana holoʻokoʻa holoʻokoʻa o nā wahine ʻimi hoʻokipa i kahi ʻano like me ka mamao o ka lele ma kahi manawa 24-hola. Ua hoʻopilikia nui ʻia ka mamao o ka lele ʻana e ka meaʻai i ʻai ʻia (F(5, 138) = 28.27, p <0.0001), a ʻo nā wahine ʻimi hoʻokipa i komo i nā hola 72 o ka mimi i hoʻohālikelike ʻia i nā meaʻai ʻē aʻe a pau. ʻOi aku ka lōʻihi o ka lele ʻana o nā quitoes ma mua o ka hou (p = 0.022) a me ka mimi 24-h-aged (p = 0.022) i hānai ʻia i nā makika. I ka hoʻohālikelike ʻana i ke ʻano hana lele i wehewehe ʻia e ka ʻai mimi, ua hōʻike nā wahine ʻimi ʻimi urea i ka hana lele hoʻomau i ka manawa 24-h, ʻike ʻia i ka manawa like o ka wahine. Ua hoʻonui nui ʻia ka mamao o ka lele ʻana o ka urea ma muli o ka hoʻopaʻa ʻia ʻana (F(5, 138) = 1310.91, p <0.0001). ʻO nā wahine ʻimi hoʻokipa i hānai ʻia i ka lele ʻana o ka urea ma mua o nā wahine i hānai i ka wai a i ʻole ka sucrose (p <0.03).
ʻO ka holo holoʻokoʻa holoʻokoʻa o nā makika ʻū koko ua kūpaʻa a mālama ʻia ma luna o 24 mau hola ma nā meaʻai āpau, me ka hoʻonui ʻana i ka hana mimi i ka hapa lua o ka wā pōʻeleʻele no nā wahine i hānai ʻia i ka wai a me nā wahine hānai hou a me 24 mau hola (kiʻi 3). ʻaʻole (F(5, 138) = 1.36, p = 0.24) .me nā mimi ʻē aʻe a me nā meaʻai hoʻomalu (hou, p = 0.0091; 72 hola, p = 0.0022; 168 hola, p = 0.001; sucrose, p = 0.0017; dH2O, 36).
Ua nānā ʻia nā hopena o ka ʻai ʻana o ka mimi a me ka urea i nā ʻāpana hānau ma nā bioassays hoʻomoe hua manu (Figure 4A) a ua noiʻi ʻia e like me ka helu o nā hua i waiho ʻia e kēlā me kēia wahine, ka nui o ka hua manu, a me nā larvae instar mua i hoʻopaʻa ʻia. ʻO ka helu o nā hua i waiho ʻia. ʻO nā wahine Arab hānai i ka urine, ʻokoʻa ʻia e ka meaʻai (F = 40.8 p. B). Ua hānai nā wahine i ka mimi 24-hola, ua ʻoi aku ka nui o ka ʻai ʻana i ke koko ma mua o nā wahine i hānai i nā meaʻai mimi ʻē aʻe a ua like me ka mea i hānai ʻia i ka sucrose (Fig. 4B). Pela nō hoʻi, ʻokoʻa ka nui o nā hua i waiho ʻia e nā wahine hānai mimi ma muli o ka meaʻai (F(5, 209) = 12.85, p <0.004 ka nui o nā hua manu i waiho ʻia ma mua o 0.004 wahine. nā wahine hānai wai, ʻoiai nā hua o nā wahine i hānai ʻia me ka 168 h o ka mimi he mea liʻiliʻi loa (Fig. 4C). Eia kekahi, ʻoi aku ka nui o ka ʻai ʻana o ka mimi i ka nui o ka larval (F(5, 187) = 7.86, p <0.0001), me ka nui o ka nui o nā larvae e puka mai ana mai nā hua i waiho ʻia e 24-hours-old7 hua i waiho ʻia e 24-hours-old7 hua manu mai ka 24-a me 24-hour. hānai a me 168-h mau wahine hānai mimi (Figure 4D).
ʻO ka hana hānau hānau o ka wahine Anopheles arabinis e hānai ana i ka mimi bipi a me ka urea. Ua hānai ʻia nā makika wahine hānai koko i nā meaʻai e pili ana i ka mimi pipi hou a ʻelemakule, nā ʻano ʻokoʻa o ka urea, sucrose (10%), a me ka wai distilled (H2O) no 48 mau hola ma mua o ka waiho ʻana i nā bioassays a me ka loaʻa ʻana o ka hua manu (48 mau hola F). a me ka nui o ka larvae (D, G) i hoʻopili nui ʻia e ka meaʻai i hāʻawi ʻia (mimi pipi: BD; urea: EG). ʻO nā ʻano no kēlā me kēia ʻāpana i ana ʻia me ka hoʻohana ʻana i nā inoa leka like ʻole he ʻokoʻa loa kekahi mai kekahi (ANOVA ala hoʻokahi me ka hoʻohana ʻana i ka loiloi post hoc a Tukey; p <0.05).
Ma ke ano he mea nui nitrogenous o ka mimi, urea, ke haawiia mai i mea ai i na wahine i hanai i ke koko, ua pili nui i na palena o ka hanau ana ma na ha'awina a pau. O ka heluna o na hua i waihoia e na wahine i hanai i ka urea, ma hope o ka ai ana i ke koko, e pili ana i ka ike o ka urea (F(11, 360) = 4.69; p < 0.0001 a me ka nui o na wahine i hanai ia ma waena o 1.0001 m. hua (Figure 4E) 1) ua ʻoi aku ka loli (Fig. 4G).
ʻO ka hoʻokipa holoʻokoʻa i ka poʻe e ʻimi nei i ka bovine mimi headspace volatile extracts.The arabiensis i helu ʻia i loko o ka olfactometer paipu aniani (Fig. 5A) ua hoʻopilikia nui ʻia e ka makahiki mimi (χ2 = 15.9, df = 4, p = 0.0032; Fig. 5B). 0.0060, 168 hola: p = 0.012, pentane: p = 0.00070), Koe naʻe ka ʻala o ka mimi hou (p = 0.13; Kiʻi 5B) . Ua ʻike ʻia he ʻoi aku ka maikaʻi o nā mea hoʻoheheʻe huluhulu i hoʻohālikelike ʻia me ka mimi kahiko 72-hola i hoʻohālikelike ʻia me nā mana (p = 0.0066; Kiʻi 5C).
Nā pane Behavioral i ka maoli a me ka synthetic pipi mimi ala i ka huli ana no ka mea hoʻokipa a me ke koko-hānai Anopheles arabianus.Schematic o ke aniani paipu olfactometer (A).Attraction o headspace volatile extracts mai ka hou a me ka elemakule bipi mimi e hookipa (B) a me ke koko-momi (C) makika. E huli i ka tentacle pane o ka Haku i.2hours (D4hours) hou. -hola (F), a me 168-hola (G) mimi bipi kahiko i hoikeia. Electron antenna detection (EAD) traces hōʻike i ka hoʻololi voltage i ka pane ana i ka bioactive compounds in the headspace eluted from the gas chromatograph and finded by flame ionization detector (FID). Hōʻike ka hōʻailona hoʻokahi (*) i ka pane haʻahaʻa haʻahaʻa like ʻole. Hōʻike nā asterisk pālua (**) i nā pane hiki ʻole ke hana hou ʻia. E ʻimi i ka mea hoʻokipa (H) a me ka mea hoʻokahe koko (I) An.arabiensis he mau manaʻo like ʻole i nā huila synthetic o nā ʻala mimi bipi hou a ʻelemakule. kālailai;p <0.05). Hōʻike nā pahu hewa i ka hewa maʻamau o ka unahi
Wahine Ann.arabiensis, 72 mau hola a me 120 mau hola ma hope o ka ʻai ʻana i ke koko, i ka wā o ka spawning, ʻaʻohe makemake i hōʻike ʻia no ke poʻo o ka mimi pipi mai ka mimi pipi hou a ʻelemakule i hoʻohālikelike ʻia me nā mana pentane (χ2 = 3.07, p> 0.05; waihona hou 1: Fig. S1).
No ka wahine Ann.arabiensis, GC-EAD a me GC-MS kānana i ʻike ʻia he ʻewalu, ʻeono, ʻekolu a me ʻekolu mau pūhui bioactive (Figure 5D-G). ua hoʻokomo ʻia i loko o nā loiloi hou aʻe.
Ua hoʻonui ʻia ka nui o ka hoʻokuʻu ʻana o nā pūhui bioactive i loko o ka hōʻiliʻili poʻo mai ka 29 µg h-1 i ka mimi hou i ka 242 µg h-1 i ka mimi 168-hola kahiko, ma muli o ka p-cresol a me ka m-formaldehyde Phenol piʻi a me ka phenol. pili i ka emi 'ana o ka hōʻailona ikaika (abundance) i loko o ka chromatogram (Fig. 5D) -G panel hema) a me ka physiological pane i kēia mau pūhui (Fig. 5D-G akau panel).
Ma keʻano holoʻokoʻa, ua like ka lakene kūlohelohe o ka hui ʻana o nā mea bioactive i ʻike ʻia i loko o nā ʻāpana volatile o nā poʻo mimi hou a ʻelemakule (Fig. 5D–G) a ʻaʻole i ʻike ʻia e hoʻopiʻi nui i ka ʻimi ʻana i kahi mea hoʻokipa (χ2 = 8.15, df = 4, p = 0.083; Fig. 5H) a i ʻole ke koko = χ2 = χ2 = 9. 0.30; Fig. 5I). Akā naʻe, ua hōʻike ʻia nā hoʻohālikelike post hoc ma waena o nā lāʻau lapaʻau he nani loa nā makika ʻimi hoʻokipa i ka hui ʻana o ka mimi 24-h mau makahiki i hoʻohālikelike ʻia me nā mana pentane (p = 0.0086; Kiʻi 5H).
E loiloi i ka hana o nā mea i pili i nāʻano synthetic ma nāʻano he nui loa, e hoʻopili ana i nāʻano likeʻole. FILE 1: Kiʻi S2A),ʻoi aku ka maikaʻi o nā hana likeʻole o ka hoʻokaʻawaleʻana i nāʻano likeʻole (0,077) 2; faile hou 1: Kuhi S2b).
Ma kahi kauhale malaria-endemic ma ʻAitiopia, ua loiloi ʻia ka maikaʻi o kahi hui synthetic o 24-hola pipi mimi i ka huki ʻana i nā makika ma lalo o nā kūlana kahua i loiloi ʻia no nā pō he ʻumi (Fig. 6A). lex spp.(Faila hou 1: Papa S1). ʻO Anopheles arabinis wale nō ka lālā o An.Gambian ʻano paʻakikī i ʻike ʻia e ka PCR analysis. Ma ka awelika, hopu ʻia nā makika 320 i kēlā me kēia pō, i loko o ia manawa ua hopu ʻia nā pahele me nā maunu hoʻohuihui synthetic i nā makika ma mua o nā pahele i hui ʻia me ka hui ʻole ʻia (χ2(0, 0.N. Ua hoʻonoho ʻia nā pahele ma kēlā me kēia o nā pō mana ʻelima i ka hoʻomaka, waena, a me ka hopena o ka hoʻokolokolo. ua hoʻonui nui ʻia: ʻimi ʻimi (χ2(0, 2107) = 138.7, p <0.0001), hānai koko hou (χ2(0, 650) = 32.2, p <0.0001) a me ka hāpai ʻana (χ2(0, 228) = 6.27, p = 0.Waihona 'ē aʻe 1: Papa S1). Hōʻike ʻia kēia ma ka huina o nā makika i hopu ʻia: ka mea hoʻokipa e ʻimi ana > hoʻoinu koko > hāpai > hāpai ʻia > kane.
Ka loiloi kahua o ka efficacy o ka 24-hola synthetic pipi mimi mea ala huikau. (C), ma ke ano okoa, he physiological state-dependent effect. Eia hou, ua hopu keia mau pahele i ka nui o ka nui o na makika Culex. 5).). Hōʻike nā hōkū i nā pae koʻikoʻi helu (*p = 0.01 a me ***p <0.0001)
Ua hopu ʻokoʻa ʻia nā ʻano ʻekolu i loko o nā pahele i loaʻa nā hui ʻana. s ua paʻa i loko o ka pahele e hoʻokuʻu ana i ka hui synthetic (Fig. 6B), ʻoiai ʻaʻole i ʻokoʻa ka nui o An. Ua ʻike ʻia ka Pharoensis ma nā ʻano physiological like ʻole (Fig. 6C). ), hoʻohālikelike ʻia me nā pahele mana.
Ua hoʻohana ʻia nā pahele maunu ma waho o nā mea hoʻokipa ma waena o nā kahua hoʻohua a me nā kaiāulu kuaʻāina o ʻAitiopia no ka loiloi inā hoʻohana nā makika malaria i ka ʻala mimi bipi ma ke ʻano he wahi noho hoʻokipa cue. a hopu ʻia, ʻoiai ma nā helu liʻiliʻi, kūʻokoʻa i ka makahiki mimi (χ2(5, 25) = 2.29, p = 0.13; waihona hou 1: Kiʻi S3 ).Ma ke ʻano like ʻole, ʻaʻole i hopu nā mana wai i nā makika malaria i nā wela kiʻekiʻe (Figure 1: Kiʻi S3).
Loaʻa a puʻunaue nā makika Malaria i nā pūhui nitrogen-containing ma o ka hānai ʻana i ka mimi pipi (ʻo ia hoʻi, nā puddles) e hoʻomaikaʻi ai i nā ʻano mōʻaukala o ke ola, e like me nā mea ʻē aʻe [2, 4, 24, 25, 26]. Loaʻa i nā makika wahine kēia punawai ma ka ʻala a hiki iā lākou ke hoʻoponopono i ka lawe ʻana o nā pūhui nitrogenous i loko o ka mimi, me ka urea, ka mea nui nitrogenous i loko o ka mimi [15, 16]. Ma muli o ke kūlana physiological o ka namu wahine, ua hoʻokaʻawale ʻia nā meaʻai i loko o ka mimi e hoʻonui ai i ka hana lele a me ke ola ʻana o nā namu wahine e ʻimi ana i ke koko a me ke ʻano o ke ola. No laila, he mea koʻikoʻi ka hui ʻana o ka mimi no nā maʻi maʻi malaria i pani ʻia e like me ka poʻe makua ʻai ʻole [8], no ka mea, hāʻawi ia i nā makika wahine i ka hiki ke loaʻa i nā pūhui nitrogenous koʻikoʻi ma o ka hana ʻana i nā meaʻai haʻahaʻa.