Hoʻohui ka loaʻa ʻana a me ka hoʻolaha ʻana i nā meaʻai i nā ʻano ʻimi pepeke a me nā ʻano mōʻaukala ola. No ka hoʻoponopono ʻana i nā hemahema i nā meaʻai kikoʻī ma nā pae ola like ʻole, hiki i nā pepeke ke loaʻa i kēia mau meaʻai ma o ka hānai ʻana i nā mea huna vertebrate i kahi hana i ʻike ʻia he puddles. Me he mea lā ua nele ka makika ʻo Anopheles arabiani i ka meaʻai a, no laila, pono nā meaʻai no ka metabolism a me ka hānau hou ʻana. ʻO ka pahuhopu o kēia noiʻi ʻana, ʻo ia ke loiloi inā hoʻomaikaʻi ka hoʻoulu ʻana o An. arabiensis ma ka mimi bipi no ka loaʻa ʻana o nā meaʻai i nā ʻano 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 168 hola, a ua ana ʻia nā wahine e ʻimi ana i ka mea hoʻokipa a hānai ʻia i ke koko (48 hola ma hope o ka ʻai ʻana i ke koko) i loko o kahi olfactometer Y-tube, a ua loiloi ʻia nā wahine hāpai no ka hoʻāʻo spawning. A laila ua hoʻohana ʻia kahi loiloi kemika a me ka electrophysiological i hui pū ʻia e ʻike i nā hui bioactive i loko o ka mimi bipi ma nā papa makahiki ʻehā. Ua loiloi ʻia nā hui synthetic o nā hui bioactive i nā hoʻokolohua Y-tube a me ke kahua. No ka noiʻi ʻana i ka mimi bipi a me kāna hui nitrogen nui i loaʻa ka urea ma ke ʻano he mau meaʻai hoʻohui no nā vectors malaria, ua ana ʻia nā ʻano hānai a me nā ʻano mōʻaukala ola. Ua loiloi ʻia ka hapa o nā makika wahine a me ka nui o ka mimi bipi a me ka urea i komo. Ma hope o ka hānai ʻana, ua loiloi ʻia nā wahine no ke ola, ka lele ʻana i hoʻopaʻa ʻia, a me ka hānau hou ʻana.
E ʻimi i ke koko a me ka meaʻai o ka mea hoʻokipa. Ma nā haʻawina hoʻokolohua a me ke kahua, ua huki ʻia nā ʻArabia i ke ʻala kūlohelohe a me ka synthetic o ka mimi bipi hou a ʻelemakule. ʻAʻole makemake nā wahine hāpai i nā pane mimi bipi ma nā wahi hānau. Hoʻopili ikaika nā wahine ʻimi kikowaena a me nā wahine omo koko i ka mimi bipi a me ka urea a hoʻokaʻawale i kēia mau kumuwaiwai e like me nā kālepa mōʻaukala ola ma ke ʻano he hana o ke kūlana physiological no ka lele ʻana, ke ola ʻana, a i ʻole ka hānau hou ʻana.
ʻO ka loaʻa ʻana a me ka hoʻolaha ʻana o ka mimi bipi no ka hoʻomaikaʻi ʻana i nā ʻano mōʻaukala ola o Anopheles arabinis. Hoʻopilikia pololei ka hānai ʻana i ka mimi bipi i ka hiki ke vector ma ka hoʻonui ʻana i ke ola o kēlā me kēia lā a me ka nui o ka vector, a ma ke ʻano ʻole ma ka hoʻololi ʻana i ka hana lele a no laila e noʻonoʻo ʻia i nā hiʻohiʻona e hiki mai ana.
Hoʻohui ka loaʻa ʻana a me ka hoʻolaha ʻana i nā meaʻai i nā ʻano ʻai pepeke a me nā ʻano mōʻaukala ola [1,2,3]. Hiki i nā pepeke ke koho a loaʻa ka meaʻai a hana i ka hānai uku ma muli o ka loaʻa ʻana o ka meaʻai a me nā koi meaʻai [1, 3]. Aia ka hoʻolaha ʻana o nā meaʻai i ke kaʻina hana mōʻaukala ola a hiki ke alakaʻi i nā koi like ʻole no ka maikaʻi a me ka nui o ka meaʻai ma nā pae ola like ʻole o nā pepeke [1, 2]. No ka hoʻoponopono ʻana i nā hemahema i nā meaʻai kikoʻī, hiki i nā pepeke ke loaʻa i kēia mau meaʻai ma o ka hānai ʻana i nā mea hoʻohui, e like me ka lepo, nā ʻano lepo like ʻole a me nā mea huna o nā vertebrates, a me ke kino make, kahi hana i ʻike ʻia he puddles [2]. ʻOiai ua wehewehe nui ʻia nā ʻano ʻano pepeke a me nā moth, loaʻa pū nā lua wai i nā kauoha pepeke ʻē aʻe, a ʻo ka hoihoi a me ka hānai ʻana i kēia mau ʻano kumuwaiwai hiki ke loaʻa nā hopena koʻikoʻi i ke olakino a me nā ʻano mōʻaukala ola ʻē aʻe [2, 4, 5, 6] ,7]. ʻO ka makika malaria ʻo Anopheles gambiae sensu lato (sl) e puka mai ana ma ke ʻano he kanaka makua 'malnourished' [8], no laila hiki i ka wai ke pāʻani i kahi kuleana koʻikoʻi i kona mau ʻano mōʻaukala ola, akā ua hoʻowahāwahā ʻia kēia ʻano a hiki i kēia manawa. ʻO ka hoʻohana ʻana i ka hoʻoulu ʻana ma ke ʻano he mea e hoʻonui ai i ka lawe ʻana i nā meaʻai i loko o kēia kaʻa koʻikoʻi e pono ai ka nānā ʻana no ka mea he hopena epidemiological koʻikoʻi paha kēia.
Ua kaupalena ʻia ka lawe ʻana o ka nitrogen i loko o nā makika wahine makua ʻo Anopheles ma muli o nā waihona calorie haʻahaʻa i lawe ʻia mai ke kahua larval a me ka hoʻohana pono ʻole ʻana i ka ʻai koko [9]. Hoʻoponopono pinepine ka wahine ʻo Ann.gambiae sl no kēia ma ka hoʻohui ʻana me nā ʻai koko hoʻohui [10, 11], no laila e hoʻokau ana i nā poʻe he nui i ka pilikia o ka loaʻa ʻana o ka maʻi a e hoʻokau ana i nā makika i ka pilikia nui o ka predation. ʻOkoʻa, hiki i nā makika ke hoʻohana i ka hānai hoʻohui o ka excreta vertebrate e loaʻa ai nā pūhui nitrogenous e hoʻonui ai i ka hoʻololi a me ka maneuverability lele, e like me ka hōʻike ʻana o nā pepeke ʻē aʻe [2]. Ma kēia ʻano, ʻo ka ikaika a me ka ʻokoʻa o kekahi o nā ʻano hoahānau i loko o An. ʻO ka paʻakikī ʻano Gambian sl, ʻo Anopheles arabinis, ka mimi bipi hou a ʻelemakule [12,13,14], he mea hoihoi. He manawa kūpono ʻo Anopheles arabinis i kāna mau makemake host a ʻike ʻia e pili pū me a hānai i nā bipi. He kumuwaiwai waiwai ka mimi bipi i nā pūhui nitrogenous, me ka urea e helu ana no 50-95% o ka huina nitrogen i loko o ka mimi hou [15, 16]. I ka wā e ʻelemakule ai ka mimi bipi, hoʻohana nā microorganisms. kēia mau kumuwaiwai e hōʻemi i ka paʻakikī o nā hui nitrogenous i loko o 24 mau hola [15]. Me ka piʻi wikiwiki ʻana o ka ammonia, e pili ana me ka emi ʻana o ka nitrogen organik, ulu nā microorganism alkalophilic (he nui o ia mau mea e hana i nā hui ʻawahia i nā makika) [15], kahi paha he wahine ʻo Ann.arabiensis makemake nui ʻia i ka mimi i piha i 24 mau hola a i ʻole [13, 14].
Ma kēia haʻawina, ua ʻimi ʻia nā Ans i hānai ʻia e ka mea hoʻokipa a me ke koko. I kona wā mua o ka gonadotropin cycle, ua loiloi ʻia nā arabiensis no ka loaʻa ʻana o nā pūhui nitrogenous, me ka urea, ma o ka hui ʻana o ka mimi. Ma hope mai, ua hana ʻia kekahi mau hoʻokolohua e loiloi pehea e hoʻokaʻawale ai nā makika wahine i kēia kumuwaiwai meaʻai no ka hoʻomaikaʻi ʻana i ke ola, ka hānau hou ʻana a me ka ʻimi ʻana hou aku. ʻO ka hope loa, ua loiloi ʻia ke ʻala o ka mimi bipi hou a me ka ʻelemakule e hoʻoholo ai inā hāʻawi kēia i nā hōʻailona hilinaʻi no ka mea hoʻokipa a me ka An i hānai ʻia e ke koko. I kā lākou ʻimi ʻana i kēia kumuwaiwai meaʻai kūpono, ua ʻike ʻo arabiensis i nā pilina kemika ma hope o ka ʻokoʻa i ʻike ʻia. Ua loiloi hou ʻia nā hui ʻala synthetic o nā pūhui organik volatile (VOCs) i ʻike ʻia i loko o ka mimi 24-hola i ʻelemakule ma lalo o nā kūlana kahua, e hoʻonui ana i nā hopena i loaʻa ma lalo o nā kūlana hoʻokolohua a hōʻike i ka hopena o ka ʻala mimi bovine ma nā kūlana physiological like ʻole. Ka huki ʻana i nā makika. Hōʻoia nā hopena i loaʻa iā An. arabiensis loaʻa a hāʻawi i nā pūhui nitrogenous i loaʻa i ka mimi vertebrate e hoʻohuli i nā ʻano mōʻaukala ola. Kūkākūkā ʻia kēia mau hopena ma ke ʻano o nā hopena epidemiological hiki a pehea e hiki ai ke hoʻohana ʻia no ka nānā ʻana a me ka kaohi ʻana i nā vector.
Ua mālama ʻia nā Anopheles arabicans (ʻano Dongola) ma 25 ± 2 °C, 65 ± 5% RH a me kahi pōʻaiapuni mālamalama:pouli 12:12 h. Ua hānai ʻia nā larvae i loko o nā pā plastik (20 cm × 18 cm × 7 cm) i hoʻopiha ʻia me ka wai i hoʻoheheʻe ʻia a hānai ʻia i ka meaʻai iʻa Tetramin® (Tetra Werke, Melle, DE). Ua hōʻiliʻili ʻia nā pupae i loko o nā kīʻaha 30 ml (Nolato Hertila, Åstorp, SE) a laila ua hoʻoili ʻia i nā hale Bugdorm (30 cm × 30 cm × 30 cm; MegaView Science, Taichung, Taiwan) e ʻae i ka puka ʻana o nā mākua. Ua hāʻawi ʻia nā mākua me kahi hopena sucrose 10% ad libitum a hiki i 4 mau lā ma hope o ka puka ʻana (dpe), ma ia manawa ua hāʻawi ʻia nā wahine e ʻimi ana i ka meaʻai ma mua koke o ka hoʻokolohua, a i ʻole ua pōloli ʻia i ka pō me ka wai i hoʻoheheʻe ʻia ma mua o ka hoʻokolohua, e like me ka mea i wehewehe ʻia ma lalo nei. Ua pōloli ʻia nā wahine i hoʻohana ʻia no nā hoʻokolohua paipu lele no 4-6 mau hola wale nō. me ka wai ad libitum. No ka hoʻomākaukau ʻana i nā makika omo koko no nā bioassays ma hope, ua hāʻawi ʻia he 4 mau wahine dpe me ke koko hipa defibrotic (Håtunalab, Bro, SE) me ka hoʻohana ʻana i kahi ʻōnaehana hānai membrane (Hemotek Discovery Workshops, Accrington, UK). A laila ua hoʻoili ʻia nā wahine i piha i ka paʻa i nā hale paʻahao pākahi a hāʻawi pololei ʻia i ka meaʻai, e like me ka mea i wehewehe ʻia ma lalo nei, a i ʻole 10% 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 ua hoʻoili ʻia i ka hale hana, a laila ua loaʻa ka wai distilled ad libitum no 4-6 mau hola ma mua o ka hoʻokolohua.
Ua hoʻohana ʻia nā hoʻāʻo hānai e helu i ka mimi a me ka ʻai ʻana o ka urea i ka wahine An.Arab makua. Ua hāʻawi ʻia nā wahine ʻimi kikowaena a me nā wahine hānai koko i kahi meaʻai e loaʻa ana he 1% ka mimi bipi hou a me ka ʻelemakule i hoʻoheheʻe ʻia, nā ʻano like ʻole o ka urea, a me ʻelua mau kaohi (10% sucrose a me ka wai) no 48 hola. Eia kekahi, ua hoʻohui ʻia ka waihoʻoluʻu meaʻai (1 mg ml-1 xylene cyanide FF; CAS 2650-17-1; Sigma-Aldrich, Stockholm, SE) i ka meaʻai a hāʻawi ʻia i loko o kahi matrix 4 × 4 i loko o nā paipu microcentrifuge 250 µl (Axygen Scientific, Union City, CA, US; Kiʻi 1A) E hoʻopiha a hiki i ka lihi (~300 µl). I mea e pale aku ai i ka hoʻokūkū ma waena o nā makika a me nā hopena hiki o ke kala dye, e kau i 10 mau makika i loko o kahi kīʻaha Petri nui (12 cm ke anawaena a me 6 cm ke kiʻekiʻe; Semadeni, Ostermundigen, CH; Kiʻi 1A) i ka pouli piha ma 25 ± 2 cm °C a me 65 ± 5% ka haʻahaʻa pili. ʻO kēia mau hoʻokolohua ua hana hou ʻia he 5 a 10 mau manawa. Ma hope o ka ʻike ʻia ʻana i ka meaʻai, ua kau ʻia nā makika ma -20 °C a hiki i ka nānā hou ʻana.
E nānā i ka mimi bipi a me ka urea i hoʻopili ʻia e ka mea hoʻokipa a me ka wahine omo koko ʻo Anopheles arabianus. I ka hoʻāʻo hānai ʻana (A), ua hāʻawi ʻia nā makika wahine me kahi meaʻai i loaʻa i ka mimi bipi hou a me ka ʻelemakule, nā ʻano like ʻole o ka urea, sucrose (10%), a me ka wai i hoʻoheheʻe ʻia (H2O). ʻOi aku ka nui o ka sucrose o nā wahine ʻimi-host (B) a me nā wahine hānai koko (C) ma mua o nā meaʻai ʻē aʻe i hoʻāʻo ʻia. E hoʻomaopopo i ka hoʻopili ʻana o nā wahine ʻimi-host i ka mimi bipi 72-hola ma lalo o ka mimi bipi 168-hola (B). Hōʻike ʻia ka awelika o ka nui o ka nitrogen (± standard deviation) o ka mimi ma ka inset. Lawe nā wahine ʻimi-host (D, F) a me nā wahine omo koko (E, G) i ka urea ma ke ʻano hilinaʻi i ka dose. ʻOkoʻa loa ka awelika o nā nui i hanu ʻia (D, E) me nā inoa leka like ʻole mai kekahi i kekahi (one-way ANOVA me ka hoʻohana ʻana i ka loiloi post hoc a Tukey; p < 0.05). Hōʻike nā kaola hewa i ka hewa maʻamau o ka awelika (BE). Hōʻike ka laina dashed pololei i ka laina regression log-linear (F, G)
No ka hoʻokuʻu ʻana i ka meaʻai i omo ʻia, ua hoʻokomo pākahi ʻia nā makika i loko o nā ʻōmole microcentrifuge 1.5 ml e loaʻa ana he 230 µl o ka wai distilled a ua hoʻopau ʻia ke kino me ka hoʻohana ʻana i kahi pestle hoʻolei a me ka motika cordless (VWR International, Lund, SE), a ukali ʻia e ka centrifugation ma 10 krpm no 10 min. Ua hoʻoili ʻia ka supernatant (200 µl) i kahi microplate 96-well (Sigma-Aldrich) a ua hoʻoholo ʻia ka absorbance (λ620) me ka hoʻohana ʻana i kahi mea heluhelu microplate e pili ana i ka spectrophotometer (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm). Ma ke ʻano ʻē aʻe, ua wili ʻia nā makika i loko o 1 ml o ka wai distilled, 900 µl o ia mea i hoʻoili ʻia i kahi cuvette no ka nānā ʻana i ka spectrophotometric (λ 620 nm; UV 1800, Shimadzu, Kista, SE). No ka helu ʻana i ka ʻai ʻana i ka meaʻai, ua hoʻomākaukau ʻia kahi curve maʻamau e ka serial dilution e loaʻa ai ka 0.2 µl a i 2.4 µl o 1 mg ml-1 xylene cyanide. A laila, ua hoʻohana ʻia ka nui o ka optical density o nā dye i ʻike ʻia e hoʻoholo ai i ka nui o ka meaʻai a kēlā me kēia makika i ʻai ai.
Ua kālailai ʻia ka ʻikepili leo me ka hoʻohana ʻana i ka loiloi hoʻokahi ala o ka variance (ANOVA) a ukali ʻia e nā hoʻohālikelike pālua post hoc a Tukey (JMP Pro, v14.0.0, SAS Institute Inc., Cary, NC, US, 1989–2007). Ua wehewehe nā loiloi regression linear i ka lawe ʻana o ka urea e pili ana i ka concentration a hoʻohālikelike i nā pane ma waena o nā makika ʻimi-host a me nā makika omo koko (GraphPad Prism v8.0.0 no Mac, GraphPad Software, San Diego, CA, US).
Ma kahi o 20 µl o nā hāpana mimi mai kēlā me kēia hui makahiki i hoʻopaʻa ʻia ma Chromosorb® W/AW (10 mg 80/100 mesh, Sigma Aldrich) a hoʻopaʻa ʻia i loko o nā capsules tin (8 mm × 5 mm). Ua hoʻokomo ʻia nā capsules i loko o ke keʻena kuni o kahi mea kālailai CHNS/O (Flash 2000, Thermo Fisher Scientific, Waltham, MA, US) e hoʻoholo i ka nui o ka nitrogen i loko o ka mimi hou a me ka mimi ʻelemakule e like me ke kaʻina hana a ka mea hana. Ua helu ʻia ka huina nitrogen (g N l-1) ma muli o nā ʻike urea i ʻike ʻia i hoʻohana ʻia ma ke ʻano he maʻamau.
No ka loiloi ʻana i ka hopena o ka ʻai ʻana i ke ola o ka wahine e ʻimi ana i ka mea hoʻokipa a me ke koko, ua hoʻonoho pākahi ʻia nā makika i loko o nā kīʻaha Petri nui (12 cm ke anawaena a me 6 cm ke kiʻekiʻe; Semadeni) me kahi puka i uhi ʻia i ka ʻupena ma ke poʻi (3 cm ke anawaena) me No ka hoʻoulu ʻana i ka ea a me ka lako meaʻai. Ua hāʻawi ʻia nā meaʻai ma hope koke o 4 dpe a ua hoʻokomo ʻia he 1% mimi bipi hou a ʻelemakule i hoʻoheheʻe ʻia, ʻehā mau ʻano o ka urea, a me ʻelua mau kaohi, 10% sucrose a me ka wai. Ua pipetted kēlā me kēia meaʻai ma luna o kahi tampon niho (DAB Dental AB, Upplands Väsby, SE) i hoʻokomo ʻia i loko o kahi syringe 5 ml (Thermo Fisher Scientific, Gothenburg, SE), wehe ʻia ka plunger, a kau ʻia ma luna o kahi kīʻaha petri (kiʻi 1).1A). E hoʻololi i kāu meaʻai i kēlā me kēia lā. E mālama i ka lab e like me ka mea i wehewehe ʻia ma luna. Ua helu ʻia nā makika ola i ʻelua manawa i ka lā, ʻoiai ua hoʻolei ʻia nā makika make a hiki i ka make ʻana o ka makika hope loa (n = 40 no kēlā me kēia lāʻau). Ua kālailai ʻia ke ola ʻana o nā makika i hānai ʻia i nā meaʻai like ʻole me ka hoʻohana ʻana i nā pihi ola ʻo Kaplan-Meyer a nā hoʻāʻo log-rank e hoʻohālikelike i nā hoʻohālikelike hoʻolaha ola ma waena o nā meaʻai (IBM SPSS Statistics 24.0.0.0).
He wili lele makika maʻamau i hoʻokumu ʻia ma Attisano et al.[17], i hana ʻia me nā panela acrylic maopopo 5 mm ka mānoanoa (10 cm ka laulā x 10 cm ka lōʻihi x 10 cm ke kiʻekiʻe) me ka ʻole o nā panela mua a me hope (Kiʻi 3: luna). He ʻākoakoa pivot me kahi paipu kū pololei i hana ʻia me ke kolamu chromatography kinoea (0.25 mm id; 7.5 cm L) me nā wēlau i hoʻopili ʻia i kahi nila pepeke i kau ʻia ma waena o kahi pālua o nā magnet neodymium 9 cm ke kaʻawale. Ua ʻoki ʻia kahi paipu ākea i hana ʻia me ka mea like (6.5 cm L) i ka paipu kū pololei e hana i kahi lima i hoʻopaʻa ʻia a me kahi lima e lawe ana i kahi ʻāpana liʻiliʻi o ka pepa alumini ma ke ʻano he hōʻailona hoʻopau kukui.
Ua hāʻawi ʻia nā wahine pōloli 24-hola i ka meaʻai i luna no 30 mau minuke ma mua o ke kāohi ʻia ʻana. A laila ua hoʻomaʻamaʻa ʻia nā makika wahine i hānai piha ʻia ma luna o ka hau no 2-3 min a hoʻopili ʻia i nā pine pepeke me ka beeswax (Joel Svenssons Vaxfabrik AB, Munka Ljungby, SE) a laila nakinaki ʻia i nā lima o nā paipu ākea. Flying Mill. Ua hoʻopaʻa ʻia nā kipi i kēlā me kēia lele e kahi mea hoʻopaʻa ʻikepili i kūkulu ʻia, a laila mālama ʻia a hōʻike ʻia me ka hoʻohana ʻana i ka polokalamu PC-Lab 2000™ (v4.01; Velleman, Gavere, BE). Ua waiho ʻia ka wili lele i loko o kahi lumi i hoʻoponopono ʻia e ke aniau (12 h:12 h, mālamalama: pouli, 25 ± 2 °C, 65 ± 5% RH).
No ka ʻike ʻana i ke ʻano o ka hana lele, ua helu ʻia ka huina mamao i lele ʻia (m) a me ka huina o nā hana lele i kēlā me kēia hola ma luna o 24 mau hola. Eia kekahi, ua hoʻohālikelike ʻia nā mamao awelika i lele ʻia e nā wahine pākahi ma waena o nā lāʻau lapaʻau a ua kālailai ʻia me ka hoʻohana ʻana i ka ANOVA hoʻokahi ala a me ka loiloi post hoc a Tukey (JMP Pro, v14.0.0, SAS Institute Inc.), kahi i manaʻo ʻia ai ka mamao awelika he loli hilinaʻi, ʻoiai ʻo ka lāʻau lapaʻau he kumu kūʻokoʻa. Eia kekahi, ua helu ʻia ka awelika o nā puni i nā ʻāpana 10-minuke.
No ka loiloi ʻana i ka hopena o ka ʻai ma ka hana hānau o An.arabiensis, ua hoʻoili pololei ʻia ʻeono wahine (4 dpe) i nā hale manu Bugdorm (30 cm × 30 cm × 30 cm) ma hope o ka hōʻiliʻili koko a laila hāʻawi i ka ʻai hoʻokolohua no 48 hola e like me ka mea i wehewehe ʻia ma luna. A laila ua wehe ʻia nā meaʻai a ua hāʻawi ʻia nā kīʻaha spawning (30 ml; Nolato Hertila) i piha i ka 20 ml o ka wai i hoʻoheheʻe ʻia i ke kolu o ka lā no 48 hola, e hoʻololi ana i kēlā me kēia 24 hola. E hana hou i kēlā me kēia papaʻai 20-50 mau manawa. Ua helu ʻia nā hua manu a hoʻopaʻa ʻia no kēlā me kēia hale manu hoʻokolohua. Ua hoʻohana ʻia nā subsamples o nā hua manu e loiloi i ka awelika o ka nui a me ka lōʻihi o nā hua manu pākahi (n ≥ 200 no kēlā me kēia ʻai) me ka hoʻohana ʻana i kahi microscope Dialux-20 (DM1000; Ernst Leitz Wetzlar, Wetzlar, DE) i lako me kahi Leica Camera (DFC) 320 R2; ʻO Leica Microsystems Ltd., DE). Ua mālama ʻia nā hua i koe i loko o kahi lumi i kāohi ʻia e ke aniau ma lalo o nā kūlana hānai maʻamau no 24 mau hola, a ua ana ʻia kahi subsample o nā larvae instar 1 i puka mai nei (n ≥ 200 no kēlā me kēia ʻai), e like me ka mea i wehewehe ʻia ma luna. Ua hoʻohālikelike ʻia ka helu o nā hua a me ka nui o nā hua a me nā larvae ma waena o nā lāʻau lapaʻau a me ka hoʻohana ʻana i ka ANOVA hoʻokahi ala a me ka loiloi post hoc a Tukey (JMP Pro, v14.0.0, SAS Institute Inc.).
Ua hōʻiliʻili ʻia nā mea hoʻoheheʻe poʻo mai ka mimi hou (1 hola ma hope o ka laʻana ʻana), 24 hola, 72 hola a me 168 hola i hoʻopaʻa ʻia mai nā laʻana i hōʻiliʻili ʻia mai nā bipi Zebu, nā lāhui Arsi. No ka maʻalahi, ua hōʻiliʻili ʻia nā laʻana mimi i ke kakahiaka nui ʻoiai nā bipi i loko o ka hale papaʻa. Ua hōʻiliʻili ʻia nā laʻana mimi mai 10 mau kānaka a ua hoʻoili ʻia he 100-200 ml o kēlā me kēia laʻana i nā ʻeke hoʻomoʻa polyamide pākahi (Toppits Cofresco, Frischhalteprodukte GmbH a me Co., Minden, DE) i loko o 3 l polyamide me ke poʻi i loko o nā pahu plastik vinyl chloride. Ua hōʻiliʻili ʻia nā mea hoʻoheheʻe poʻo mai kēlā me kēia laʻana mimi bipi pololei (hou) a i ʻole ma hope o ka oʻo ʻana ma ka mahana o ka lumi no 24 hola, 72 hola a me 168 hola, ʻo ia hoʻi, ua kū kēlā me kēia laʻana mimi no kēlā me kēia hui makahiki.
No ka hōʻiliʻili ʻana o nā mea hoʻoheheʻe poʻo, ua hoʻohana ʻia kahi ʻōnaehana pani e hoʻohele i kahi kahawai kinoea i kānana ʻia i ke kalapona (100 ml min-1) ma o kahi ʻeke polyamide i ke kolamu adsorption no 2.5 h ma ka hoʻohana ʻana i kahi pamu vacuum diaphragm (KNF Neuberger, Freiburg, DE). Ma ke ʻano he kaohi, ua hana ʻia ka hōʻiliʻili ʻana o ka headspace mai kahi ʻeke polyamide hakahaka. Ua hana ʻia ke kolamu adsorption me ka paipu Teflon (5.5 cm x 3 mm id) e loaʻa ana he 35 mg o Porapak Q (50/80 mesh; Waters Associates, Milford, MA, US) ma waena o nā plugs huluhulu aniani. Ma mua o ka hoʻohana ʻana, ua holoi ʻia ke kolamu me 1 ml n-hexane i hoʻoheheʻe hou ʻia (Merck, Darmstadt, DE) a me 1 ml pentane (99.0% solvent maʻemaʻe GC grade, Sigma Aldrich). Ua eluted ʻia nā mea hoʻoheheʻe i hoʻopili ʻia me 400 μl o pentane. Ua hōʻiliʻili ʻia nā hōʻiliʻili poʻo a laila mālama ʻia ma -20°C a hiki i ka hoʻohana ʻia ʻana no ka nānā hou aku.
Nā pane ʻano o ka ʻimi kikowaena a me ka ʻai koko ʻana o An. Ua kālailai ʻia nā ʻāpana volatile headspace i hōʻiliʻili ʻia mai ka mimi hou, 24-h, 72-h, a me 168-h-makahiki no nā ʻāpana volatile mai nā makika Arabidopsis me ka hoʻohana ʻana i kahi olfactometer paipu aniani pololei [18]. Ua hana ʻia nā hoʻokolohua i ka wā o ZT 13-15, ka wā kiʻekiʻe o ka hana ʻimi home a An. Arab [19]. Ua hoʻomālamalama ʻia kahi olfactometer paipu aniani (80 cm × 9.5 cm id) me 3 ± 1 lx o ke kukui ʻulaʻula mai luna mai. Ua kānana ʻia ke kahe ea i kānana ʻia a hoʻomaʻū ʻia (25 ± 2 °C, 65 ± 2% humidity pili) i hala i ka bioassay ma 30 cm s-1. Hoʻouna ʻia ka ea ma o kahi moʻo o nā pale mesh kila kila, e hana ana i kahi kahe laminar a me kahi ʻano plume like. ʻO ka mea hoʻopuka tampon niho (4 cm × 1 cm; L:D; DAB Dental AB), i kau ʻia mai kahi ʻōwili 5 cm ma ka ʻaoʻao makani o ka olfactometer, me nā hoʻololi stimulator i kēlā me kēia 5 mau minuke. No ka nānā ʻana, ua hoʻohana ʻia he 10 μl o kēlā me kēia extract headspace, i hoʻoheheʻe ʻia he 1:10, ma ke ʻano he stimulus. Ua hoʻohana ʻia ka nui like o ka pentane ma ke ʻano he kaohi. Ua hoʻokomo ʻia nā makika ʻimi kikowaena a i ʻole nā ​​​​makika omo koko i loko o nā hale hoʻokuʻu pākahi 2-3 mau hola ma mua o ka hoʻomaka ʻana o ka hoʻokolohua. Ua kau ʻia ka hale hoʻokuʻu ma ka ʻaoʻao makani o ka olfactometer, a ua ʻae ʻia nā makika e hoʻomaʻamaʻa no 1 min, a laila ua wehe ʻia ka valve butterfly o ka hale e hoʻokuʻu. Ua kālailai ʻia ka hoihoi i ka mālama ʻana a i ʻole ka kaohi e like me ka hapa o nā makika i hoʻopili ʻia me ke kumu i loko o 5 mau minuke o ka hoʻokuʻu ʻana. Ua hana hou ʻia kēlā me kēia headspace volatile extract a me ka kaohi ma ka liʻiliʻi he 30 mau manawa, a no ka pale ʻana i nā hopena o kekahi lā hoʻokahi, ua hoʻāʻo ʻia ka helu like o nā lāʻau a me nā kaohi i kēlā me kēia lā hoʻokolohua. E ʻimi i nā pane mai ka mea hoʻokipa a me nā Ans i hānai ʻia i ke koko. Ua kālailai ʻia nā set Arabic versus headspace me ka hoʻohana ʻana i ka regression logistic nominal i ukali ʻia e nā hoʻohālikelike pairwise no nā lakio ʻē (JMP Pro, v14.0.0, SAS Institute Inc.).
ʻO ka pane hānau ʻana o An. Ua kālailai ʻia nā ʻāpana headspace mai ka mimi bipi hou a ʻelemakule i loko o nā pā Bugdorm (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 i hoʻoheheʻe ʻia i hāʻawi i ka substrate hānau a ua kau ʻia ma nā kihi ʻē aʻe o ka pā, 24 cm ke kaʻawale. Ua hoʻoponopono ʻia nā kīʻaha lapaʻau me 10 μl o kēlā me kēia ʻāpana headspace ma kahi dilution 1:10. Ua hoʻohana ʻia ka nui like o ka pentane e hoʻoponopono i ke kīʻaha kaohi. Ua hoʻololi ʻia nā kīʻaha lapaʻau a me nā kīʻaha kaohi ma waena o kēlā me kēia hoʻokolohua e kaohi no nā hopena kūlana. Ua hoʻokuʻu ʻia he ʻumi mau wahine hānai koko i loko o nā pā hoʻokolohua ma ZT 9-11 a ua helu ʻia nā hua i loko o nā kīʻaha 24 mau hola ma hope. ʻO ke ʻano no ka helu ʻana i ka helu hānau: (ka helu o nā hua i waiho ʻia i loko o ke kīʻaha lapaʻau - ka helu o nā hua i waiho ʻia i loko o ke kīʻaha kaohi) / (ka huina o nā hua i waiho ʻia). Ua hana hou ʻia kēlā me kēia lapaʻau i 8 mau manawa.
Ua hana ʻia ka nānā ʻana o 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]. I ka pōkole, ua hoʻokaʻawale ʻia nā ʻāpana volatile headspace hou me ka hoʻohana ʻana i kahi Agilent Technologies 6890 GC (Santa Clara, CA, US) i lako me kahi kolamu HP-5 (30 m × 0.25 mm id, 0.25 μm ka mānoanoa o ka ʻili, Agilent Technologies). a me ka mimi ʻelemakule. Ua hoʻohana ʻia ka hydrogen ma ke ʻano he pae mobile me ka awelika o ke kahe linear o 45 cm s-1. Ua hoʻokomo ʻia kēlā me kēia laʻana (2 μl) no 30 kekona ma ke ʻano splitless me ka mahana inlet o 225 °C. Ua hoʻolālā ʻia ka mahana o ka umu GC mai 35 °C (3 minuke paʻa) a i 300 °C (10 minuke paʻa) ma 10 °C min-1. I loko o ka GC effluent splitter, ua hoʻohui ʻia he 4 psi o ka nitrogen a ua māhele ʻia 1:1 i loko o kahi Gerstel 3D/2 low dead volume cross (Gerstel, Mülheim, DE) ma waena o ka mea ʻike lapalapa ionization a me ka EAD. Ua hoʻouna ʻia ke capillary GC effluent no EAD ma o kahi laina hoʻoili Gerstel ODP-2, kahi e hahai ai i ka mahana o ka umu GC me 5 °C, i loko o kahi paipu aniani (10 cm × 8 mm), kahi i hui pū ʻia ai me ka ea kānana kalapona, humidified (1.5 l min−1). Ua kau ʻia ka antenna 0.5 kenimika mai ka puka o ka paipu. Hoʻokahi kope i hana ʻia no kēlā me kēia makika pākahi, a no nā makika e ʻimi ana i ka mea hoʻokipa, ua hana ʻia ma ka liʻiliʻi he ʻekolu mau kope ma nā hāpana mimi o kēlā me kēia makahiki.
ʻIke ʻana i nā hui bioactive i nā hōʻiliʻili headspace o ka mimi bipi hou a me ka ʻelemakule me ka hoʻohana ʻana i ka GC i hui pū ʻia a me ka spectrometer mass (GC-MS; 6890 GC a me 5975 MS; Agilent Technologies) e hoʻopuka i nā pane antennal i ka loiloi GC-EAD, e hana ana ma ke ʻano ionization hopena electron ma 70 eV. Ua hoʻolako ʻia ka GC me kahi kolamu capillary silica fused HP-5MS UI (60 m × 0.25 mm diameter inner, 0.25 μm film thickness) me ka hoʻohana ʻana i ka helium ma ke ʻano he mobile phase me ka awelika linear flow rate o 35 cm s-1. Ua hoʻokomo ʻia kahi laʻana 2 μl me ka hoʻohana ʻana i nā hoʻonohonoho injector like a me ka mahana umu e like me ka loiloi GC-EAD. Ua ʻike ʻia nā hui ma muli o ko lākou manawa paʻa (Kovát index) a me nā spectra mass i hoʻohālikelike ʻia me ka waihona puke maʻamau a me ka waihona puke NIST14 (Agilent). Ua hōʻoia ʻia nā hui i ʻike ʻia ma ka hoʻokomo ʻana i nā kūlana maoli (Additional File 1: Table S2). No ka quantification, heptyl acetate (10 ng, 99.8% maʻemaʻe kemika, Aldrich) ua hoʻokomo ʻia ma ke ʻano he maʻamau o waho.
Ke loiloi nei i ka pono o kahi hui ʻala synthetic i haku ʻia me nā pūhui bioactive i ʻike ʻia i loko o ka mimi hou a me ka mimi ʻelemakule e huki i ka mea ʻimi kikowaena a me ka mea omo koko ʻo Ans.arabiensis, me ka hoʻohana ʻana i ka olfactometer like a me ke kaʻina hana e like me luna. Ua hoʻohālike nā hui synthetic i ka haku mele a me nā ʻāpana o nā pūhui i loko o nā ʻāpana volatile headspace i hui ʻia o ka mimi hou, 24-hola, 48-hola, 72-hola, a me 168-hola (Kiʻi 5D-G; Waihona Hou 1: Papa S2). No ka nānā ʻana, e hoʻohana i ka 10 μl o kahi dilution 1:100 o ka hui synthetic piha, me ka helu hoʻokuʻu holoʻokoʻa mai kahi o 140-2400 ng h-1, no ka loiloi ʻana i ka hoihoi i ka mea hoʻokipa a me nā makika omo koko. Ma hope o kēlā, hana ʻia ka hoʻāʻo ma nā hui piha, kahi e wehe ʻia ai nā hui subtractive o nā pūhui hoʻokahi o ka hui piha. E ʻimi i nā pane mai ka mea hoʻokipa a me ka Ans. Ua kālailai ʻia nā hui Arab vs synthetic a me subtractive me ka hoʻohana ʻana i ka regression logistic nominal i ukali ʻia e nā hoʻohālikelike pairwise no nā lakio ʻē. (JMP Pro, v14.0.0, SAS Institute Inc.).
No ka loiloi ʻana inā hiki i ka mimi bipi ke lawelawe ma ke ʻano he hōʻailona no ka noho ʻana o nā makika malaria, ua hoʻiliʻili ʻia ka mimi bipi hou a me ka ʻelemakule, i hōʻiliʻili ʻia e like me ka mea i wehewehe ʻia ma luna, a ua waiho ʻia ka wai i loko o nā bākeke 3 lita i hoʻopaʻa ʻia (100 ml) a kau ʻia i loko o nā pahele maunu kikowaena. (Mana BG-HDT; BioGents, Regensburg, DE). He ʻumi mau pahele i kau ʻia he 50 m ka mamao ma kahi hānai holoholona, ​​400 m mai ke kaiāulu kauhale (Silay, ʻAitiopa, 5°53´24´´N, 37°29´24´´E) a ʻaʻohe pipi, ma nā wahi hānau mau a me nā kauhale. Ua hoʻomehana ʻia ʻelima mau pahele e hoʻohālike i ke alo o kahi kikowaena, ʻoiai ua waiho ʻia ʻelima mau pahele me ka hoʻomehana ʻole ʻia. Hoʻololi ʻia kēlā me kēia wahi mālama i kēlā me kēia pō no ka huina o ʻelima mau pō. Ua hoʻohālikelike ʻia ka helu o nā makika i hopu ʻia i loko o nā pahele i hoʻopaʻa ʻia me ka mimi o nā makahiki like ʻole me ka hoʻohana ʻana i ka regression logistic me kahi beta binomial distribution (JMP Pro, v14.0.0, SAS Institute Inc.).
Ma ke kauhale i loaʻa i ka maʻi malaria kokoke i ke kūlanakauhale ʻo Maki, ʻāpana ʻo Oromia, ʻAitiopa (8° 11′ 08″ N, 38° 81′ 70″ E; Kiʻi 6A). Ua mālama ʻia ke aʻo ʻana ma waena o waena o ʻAukake a me waena o Kepakemapa ma mua o ka pīpī koena makahiki i loko o ka hale, me ke kau ua lōʻihi. ʻElima mau pālua o nā hale (20-50 m ke kaʻawale) aia ma waho o ke kauhale i koho ʻia no ke aʻo ʻana (Kiʻi 6A). ʻO nā pae hoʻohālikelike i hoʻohana ʻia no ke koho ʻana i nā hale: ʻaʻohe holoholona i ʻae ʻia i loko o ka hale, ʻaʻohe kuke ʻana i loko (e huki ana i ka wahie a i ʻole ke kalapona) i ʻae ʻia (ma ka liʻiliʻi i ka wā hoʻāʻo), a me nā hale me ka nui o ʻelua mau kamaʻāina, e hiamoe ana i nā mea pepehi pepeke ʻole. ma lalo o ka ʻupena makika i mālama ʻia. Ua hāʻawi ʻia ka ʻae kūpono e ka Papa Hoʻoponopono Noiʻi Hoʻonaʻauao (IRB/022/2016) o ke Kulanui o nā ʻEpekema Kūlohelohe (CNS-IRB), ke Kulanui ʻo Addis Ababa, e like me nā alakaʻi i hoʻokumu ʻia e ka World Medical Association Declaration of Helsinki. Ua loaʻa ka ʻae mai kēlā me kēia poʻo o ka ʻohana me ke kōkua o nā limahana hoʻonui olakino. Ua ʻae ʻia ke kaʻina holoʻokoʻa e nā hoʻokele kūloko ma ka pae ʻāpana a me ka ward ('kebele'). Ua hahai ka hoʻolālā hoʻokolohua i kahi hoʻolālā huinaha Latin 2 × 2, kahi i hāʻawi ʻia ai nā hui synthetic a me nā kaohi i nā hale i hui pū ʻia i ka pō mua a ua hoʻololi ʻia ma waena o nā hale i ka pō hoʻokolohua aʻe. Ua hana hou ʻia kēia kaʻina hana he ʻumi manawa. Eia kekahi, no ka helu ʻana i ka hana makika ma nā hale i koho ʻia, ua hoʻonohonoho ʻia nā pahele CDC e holo i ʻelima mau pō i ka hoʻomaka, waena a me ka hopena o ka hoʻokolohua kahua i ka manawa like o ka lā.
Ua hoʻoheheʻe ʻia kahi hui synthetic e loaʻa ana nā hui bioactive ʻeono 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 kahi dispenser wick pulupulu [20]. Ua ʻae ka dispenser wick i nā hui āpau e hoʻokuʻu ʻia ma nā ʻano like ʻole ma o ka hoʻokolohua 12 hola. Ua hoʻohana ʻia ʻo Heptane ma ke ʻano he kaohi. Ua kau ʻia ka vial ma kahi kokoke i ke komo ʻana o ka Centers for Disease Control and Prevention (CDC) light trap (John W. Hock Company, Gainesville, FL, US; Kiʻi 6A). Ua kau ʻia nā pahele 0.8 - 1 m ma luna o ka lepo, kokoke i ka wāwae o ka moena, a ua hiamoe kekahi mea manawaleʻa ma lalo o kahi ʻupena makika i mālama ʻole ʻia a ua hana ʻia ma waena o 18:00 a me 06:30. Ua nānā ʻia nā makika i hopu ʻia e ke kāne a me ke kūlana physiological (ʻaʻohe hānai, hānai ʻia, hapa hāpai, a hāpai [21] ma hope o ka nānā ʻana me ka hoʻohana ʻana i ka loiloi polymerase chain reaction (PCR) e ʻike i nā ʻano i ʻike ʻia ma ke ʻano he A. gambiae sl. Nā lālā o ka paʻakikī [23]. Ma ke kahua Ma ke aʻo ʻana, ua kālailai ʻia ka pahele ʻana o nā hale i hui pū ʻia me ka hoʻohana ʻana i kahi kumu hoʻohālike logistic kūpono, kahi i lilo ai ka hoihoi i mea hilinaʻi a ʻo ka mālama ʻana (hui synthetic vs control) ka hopena paʻa (JMP® 14.0. 0. SAS Institute Inc.). Maanei, hōʻike mākou i nā waiwai χ2 a me nā p-values ​​​​mai ka hoʻāʻo likelihood ratio.
E loiloi inā palekana. Ua hiki iā arabiensis ke loaʻa ka mimi, kona kumu naikokene nui, urea, ma o ka hānai pololei ʻana, i loko o 48 hola o ka lawelawe ʻana no 4 mau lā ma hope o (dpe) nā hoʻokolohua hānai wahine ʻimi kikowaena a me ka hānai koko (Kiʻi 1A). ʻOi aku ka nui o ka sucrose i omo ʻia e nā wahine ʻimi kikowaena a me nā wahine omo koko ma mua o nā meaʻai a wai ʻē aʻe (F(5,426) = 20.15, p < 0.0001 a me F(5,299) = 56.00, p < 0.0001, kēlā me kēia; Kiʻi 1B,C). Eia kekahi, ʻoi aku ka liʻiliʻi o ka ʻai ʻana o nā wahine ʻimi kikowaena i loko o ka mimi i 72 mau hola i hoʻohālikelike ʻia me ka mimi i 168 mau hola (Kiʻi 1B). I ka wā i hāʻawi ʻia ai kahi meaʻai me ka urea, ua omo nā wahine ʻimi kikowaena i ka nui o ka urea ma 2.69 mM i hoʻohālikelike ʻia me nā ʻano ʻē aʻe a me ka wai, ʻoiai ʻaʻole hiki ke hoʻokaʻawale ʻia mai 10% sucrose (F(10,813) = 15.72, p < 0.0001; Kiʻi 1D). He ʻokoʻa kēia i ka pane a nā wahine i hānai ʻia i ke koko, ka mea i hoʻopili pinepine i nā meaʻai i loaʻa ka urea ma mua o ka wai, ʻoiai ʻoi aku ka liʻiliʻi ma mua o 10% sucrose (F(10,557) = 78.35, p < 0.0001; Kiʻi 1).1E). Eia kekahi, i ka hoʻohālikelike ʻana ma waena o nā kūlana physiological ʻelua, ua hoʻopili nā wahine phlebotomized i ka urea ma mua o nā wahine e ʻimi ana i ka host ma nā ʻano haʻahaʻa loa, a ua hoʻopili kēia mau wahine i nā nui like o ka urea ma nā ʻano kiʻekiʻe (F(1,953)= 78.82, p < 0.0001; Kiʻi 1F, G). ʻOiai ʻo ka lawe ʻana mai kahi meaʻai i loaʻa ka urea me nā waiwai kūpono (Kiʻi 1D, E), ua hiki i nā wahine ma nā kūlana physiological ʻelua ke hoʻololi i ka nui o ka urea i hoʻopili ʻia ma luna o ka laulā holoʻokoʻa o nā ʻano urea ma ke ʻano log-linear (Kiʻi 1F, G). ). Pēlā nō, ʻike ʻia nā makika e kāohi i kā lākou lawe ʻana i ka naikokene ma ka hoʻoponopono ʻana i ka nui o ka mimi i omo ʻia, ʻoiai ʻo ka nui o ka naikokene i loko o ka mimi e hōʻike ʻia ana i ka nui i omo ʻia (Kiʻi 1B, C a me B insets).
No ka loiloi ʻana i nā hopena o ka mimi a me ka urea i ke ola ʻana o ka makika ʻimi-hoʻokipa a me ka namu omo koko, ua hānai ʻia nā wahine i ka mimi o nā makahiki ʻehā (hou, 24 hola, 72 hola, a me 168 hola ma hope o ka waiho ʻana) a me kahi ʻano o nā urea, a me ka wai i hoʻoheheʻe ʻia a me ka 10% sucrose i lawelawe ʻia ma ke ʻano he kaohi (Kiʻi 2A). Ua hōʻike kēia loiloi ola ʻana he hopena koʻikoʻi ka ʻai i ke ola holoʻokoʻa i nā wahine ʻimi-hoʻokipa (mimi: χ2 = 108.5, df = 5, p < 0.0001; urea: χ2 = 122.8, df = 5, p < 0.0001; Kiʻi 2B, C) a me nā wahine hānai koko (mimi: χ2 = 93.0, df = 5, p < 0.0001; urea: χ2 = 137.9, df = 5, p < 0.0001; Kiʻi 2D, E). Ma nā mea āpau. I loko o nā hoʻokolohua, ua emi loa ka nui o ke ola ʻana o nā wahine i hānai ʻia i ka mimi, urea, a me ka wai i hoʻohālikelike ʻia me nā wahine i hānai ʻia i ka ʻai sucrose (Kiʻi 2B-E). Ua hōʻike nā wahine ʻimi kikowaena i hānai ʻia i ka mimi hou a me ka mimi kahiko i nā helu ola like ʻole, me ka poʻe i hānai ʻia i ka mimi kahiko 72-h (p = 0.016) i loaʻa ka haʻahaʻa loa o ke ola (Kiʻi 2B). Eia kekahi, ʻo nā wahine ʻimi kikowaena i hānai ʻia i 135 mM urea i ola lōʻihi ma mua o nā kaohi wai (p < 0.04) (Kiʻi 2C). I ka hoʻohālikelike ʻia me ka wai, ua ola lōʻihi nā wahine i hānai ʻia me ka mimi hou a me ka mimi 24-hola (p = 0.001 a me p = 0.012, kēlā me kēia; Kiʻi 2D), ʻoiai ʻo nā wahine i hānai ʻia me ka mimi 72-hola i ola lōʻihi ma mua o ka poʻe i hānai ʻia i ka mimi hou pōkole wahine a me ka mimi kahiko 24-hola (p < 0.0001 a me p = 0.013, kēlā me kēia; Kiʻi 2D). I ka wā i hānai ʻia ai i 135 mM urea, ua ola lōʻihi nā wahine i hānai ʻia i ke koko ma mua o nā ʻano ʻē aʻe o ka urea a me ka wai (p < 0.013; Kiʻi 2E).
Ke ola ʻana o ka wahine 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 i loaʻa i ka mimi bipi hou a me ka mimi ʻelemakule, nā ʻano like ʻole o ka urea, sucrose (10%) a me ka wai i hoʻoheheʻe ʻia (H2O). Ua hoʻopaʻa ʻia ke ola ʻana o nā makika e ʻimi ana i ka mea hānai (B, C) a me nā makika e omo ana i ke koko (D, E) i kēlā me kēia 12 hola a hiki i ka wā e make ai nā wahine āpau i hānai ʻia i ka mimi (B, D) a me ka urea (C, E), a me nā kaohi, Sucrose a me ka wai.
ʻOkoʻa ka mamao holoʻokoʻa a me ka helu o nā puni i hoʻoholo ʻia ma ka hoʻāʻo wili lele ma luna o kahi manawa 24-hola ma waena o nā makika ʻimi kikowaena a me nā makika omo koko, kahi i hōʻike i ka hana lele liʻiliʻi ma ke ʻano holoʻokoʻa (Kiʻi 3). ʻO nā makika ʻimi kikowaena i hāʻawi i ka mimi hou a ʻelemakule a i ʻole sucrose a me ka wai i hōʻike i nā ʻano lele like ʻole (Kiʻi 3), me nā wahine e hānai ana i ka mimi hou e ʻoi aku ka ikaika i ka wanaʻao, ʻoiai ʻo ka poʻe i hānai ʻia i 24- a me 168-hola mau makika i hānai ʻia i ka mimi i hōʻike i nā ʻano lele like ʻole a he diurnal nui. ʻO nā makika wahine i hāʻawi i ka sucrose a i ʻole 72-hola mimi i hōʻike i ka hana ma loko o ka manawa 24-hola, ʻoiai ʻo nā wahine i hāʻawi i ka wai ua ʻoi aku ka ikaika i ka wā waena. ʻO nā makika i hānai ʻia i ka sucrose i hōʻike i nā pae kiʻekiʻe loa o ka hana i ka pō a me ke kakahiaka nui, ʻoiai ʻo ka poʻe i ʻai i ka mimi 72-hola i ʻike i ka emi mau o ka hana ma luna o 24 mau hola (Kiʻi 3).
ʻO ka hana lele o ka wahine Anopheles arabinis e ʻimi ana i ke koko e hānai ana i ka mimi bipi a me ka urea. I ka hoʻāʻo ʻana i ka wili lele, ua hoʻopaʻa ʻia nā makika wahine i hānai ʻia i ka mimi bipi hou a me ka ʻelemakule, nā ʻano like ʻole o ka urea, sucrose (10%), a me ka wai i hoʻoheheʻe ʻia (H2O) i nā lima ākea, e wili manuahi ana (ma luna). No nā wahine e ʻimi ana i ka kikowaena (hema) a me ke koko (ʻākau), ua hoʻopaʻa ʻia ka mamao holoʻokoʻa a me ka helu o nā lele i kēlā me kēia hola no kēlā me kēia ʻai ma luna o 24 mau hola (ʻeleʻele: hina; mālamalama: keʻokeʻo). Hōʻike ʻia ka mamao awelika a me ka helu awelika o nā hakakā ma ka ʻākau o ka pakuhi hana circadian. Hōʻike nā kaha hewa i ka hewa maʻamau o ka awelika. ʻIke i ka loiloi helu. ʻIke i ka kikokikona
Ma keʻano laulā, ua hahai ka hana lele holoʻokoʻa o nā wahine ʻimi kikowaena i kahi ʻano like me ka mamao lele ma luna o 24 mau hola. Ua hoʻopilikia nui ʻia ka mamao lele awelika e ka meaʻai i ʻai ʻia (F(5, 138) = 28.27, p < 0.0001), a ʻo nā wahine ʻimi kikowaena i ʻai ʻia he 72 mau hola o ka mimi i lele i nā mamao lōʻihi loa i hoʻohālikelike ʻia me nā meaʻai ʻē aʻe a pau (p < 0.0001), a ʻoi aku ka lōʻihi o ka lele ʻana o nā makika i hānai ʻia me ka sucrose ma mua o nā makika hou (p = 0.022) a me nā makika i hānai ʻia me ka mimi 24-h (p = 0.022). I ka hoʻohālikelike ʻana i ke ʻano hana lele i wehewehe ʻia e ka ʻai mimi, ua hōʻike nā wahine ʻimi kikowaena i hānai ʻia me ka urea i ka hana lele mau ma luna o 24 mau hola, e piʻi ana i ka hapa lua o ka wā pōʻeleʻele (Kiʻi 3). ʻOiai ua like nā ʻano hana, ua hoʻonui nui nā wahine ʻimi kikowaena i hānai ʻia me ka urea i ka mamao lele awelika ma muli o ka nui o ka mea i hoʻopili ʻia (F(5, 138) = 1310.91, p < 0.0001). ʻO ka ʻimi kikowaena ʻO nā wahine i hānai ʻia i kekahi ʻano urea i lele ʻoi aku ka lōʻihi ma mua o nā wahine i hānai ʻia i ka wai a i ʻole ka sucrose (p < 0.03).
Ua paʻa ka hana lele holoʻokoʻa o nā makika omo koko a ua hoʻomau ʻia ma luna o 24 mau hola ma nā ʻai āpau, me ka hoʻonui ʻia o ka hana mimi i ka hapa lua o ka wā pōʻeleʻele no nā wahine i hānai ʻia i ka wai a me nā wahine i hānai ʻia i ka hou a me 24 mau hola (kiʻi 3). ʻOiai ua hoʻopilikia nui ka ʻai mimi i ka mamao lele awelika i nā wahine i hānai ʻia i ke koko (F(5, 138) = 4.83, p = 0.0004), ʻaʻole i hana ka ʻai urea (F(5, 138) = 1.36, p = 0.24) me nā mimi ʻē aʻe a me ka ʻai kaohi (hou, p = 0.0091; 72 hola, p = 0.0022; 168 hola, p = 0.001; sucrose, p = 0.0017; dH2O, p = 0.036).
Ua loiloi ʻia nā hopena o ka mimi a me ka hānai ʻana o ka urea i nā ʻano hana hānau ma nā bioassays e waiho ana i nā hua manu (Kiʻi 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, a me nā larvae instar mua i puka hou ʻia. ʻO ka helu o nā hua i waiho ʻia. ʻOkoʻa nā wahine Arab i hānai ʻia i ka mimi ma muli o ka ʻai (F(5,222) = 4.38, p = 0.0008; Kiʻi 4B). ʻOi aku ka nui o nā hua i hānai ʻia e nā wahine i ka mimi 24 hola, a ua like lākou me nā mea i hānai ʻia me ka sucrose (Kiʻi 4B). Pēlā nō, ʻokoʻa ka nui o nā hua i waiho ʻia e nā wahine i hānai ʻia i ka mimi ma muli o ka ʻai (F(5, 209) = 12.85, p < 0.0001), me ka mimi 24 hola a me nā wahine i hānai ʻia me ka sucrose e waiho ana i nā hua nui aʻe ma mua o nā wahine i hānai ʻia i ka wai, ʻoiai ʻoi aku ka liʻiliʻi o nā hua o nā wahine i hānai ʻia me 168 h o ka mimi (Kiʻi 4C). Eia kekahi, ua hoʻopilikia nui ka ʻai mimi i ka nui o ka larval (F(5, 187) = 7.86, p < 0.0001), me nā larvae nui aʻe e puka mai ana mai nā hua i waiho ʻia e nā wahine hānai mimi he 24 a me 72 hola ma mua o nā hua i waiho ʻia mai nā larvae hua. ʻO nā wahine hānai wai a me nā wahine hānai mimi he 168 hola (Kiʻi 4D).
ʻO ka hana hānau o nā wahine Anopheles arabinis e hānai ana i ka mimi bipi a me ka urea. Ua hānai ʻia nā makika wahine i hānai ʻia i ke koko i nā meaʻai i loko o ka mimi bipi hou a me ka mimi bipi ʻelemakule, nā ʻano like ʻole o ka urea, sucrose (10%), a me ka wai i hoʻoheheʻe ʻia (H2O) no 48 mau hola ma mua o ke kau ʻana i nā bioassays a me ka loaʻa ʻana o nā substrates e waiho ana i nā hua manu 48 mau hola (A). Ua hoʻopilikia nui ʻia ka helu hua (B, E), ka nui o ka hua manu (C, F) a me ka nui o ka larvae (D, G) e ka meaʻai i hāʻawi ʻia (mimi bipi: BD; urea: EG). ʻO ke ʻano no kēlā me kēia parameter i ana ʻia me ka hoʻohana ʻana i nā inoa leka like ʻole he ʻokoʻa loa kekahi i kekahi (ONE-WAI ANOVA me ka hoʻohana ʻana i ka loiloi post hoc a Tukey; p < 0.05). Hōʻike nā kaola hewa i ka hewa maʻamau o ka awelika
Ma ke ʻano he ʻāpana nitrogenous nui o ka mimi, ʻo ka urea, i ka wā i hāʻawi ʻia ai ma ke ʻano he meaʻai i nā wahine i hānai ʻia i ke koko, ua hoʻopilikia nui i nā ʻano hānau i nā haʻawina āpau. ʻO ka nui o nā hua i waiho ʻia e nā wahine i hānai ʻia i ka urea, ma hope o ka ʻai ʻana i ke koko, ma muli o ka nui o ka urea (F(11, 360) = 4.69; p < 0.0001), ʻo nā wahine i hānai ʻia i nā nui o ka urea ma waena o 134 µM a me 1.34 mM i waiho i nā hua hou aʻe (Kiʻi 4E). ʻO nā wahine i hānai ʻia i nā nui o ka urea o 134 µM a i ʻole ma luna e waiho ana i nā hua nui aʻe ma mua o nā wahine i hānai ʻia i ka wai (F(10, 4245) = 36.7; p < 0.0001; Kiʻi 4F), a ʻo ka nui o ka larval, ʻoiai ua hoʻopilikia ʻia e nā nui like o ka urea i nā makuahine (F(10, 3305) = 37.9; p < 0.0001) ua ʻoi aku ka loli (Kiʻi 4G).
ʻO ka hoihoi holoʻokoʻa i nā mea hoʻokahe monile i ka ʻōpū mimi bovine e ʻimi nei i ka host. Ua hoʻopilikia nui ʻia ka arabiensis i loiloi ʻia ma ka olfactometer paipu aniani (Kiʻi 5A) e ka makahiki mimi (χ2 = 15.9, df = 4, p = 0.0032; Kiʻi 5B). Ua hōʻike ka loiloi Post hoc ua hoʻoulu ka ʻala mimi stale i 24 mau hola i nā pae kiʻekiʻe o ka hoihoi i hoʻohālikelike ʻia me nā lāʻau lapaʻau ʻē aʻe (72 hola: p = 0.0060, 168 hola: p = 0.012, pentane: p = 0.00070), Koe wale nō ke ʻala o ka mimi hou (p = 0.13; Kiʻi 5B). ʻOiai ʻaʻole ʻokoʻa loa ka hoihoi holoʻokoʻa o nā makika omo koko i ka ʻala mimi (χ2 = 8.78, df = 4, p = 0.067; Kiʻi 5C), ua ʻike ʻia kēia mau wahine he ʻoi aku ka hoihoi i nā mea hoʻokahe monile headspace i hoʻohālikelike ʻia me ka mimi 72 hola i hoʻohālikelike ʻia me nā kaohi (p = 0.0066; Kiʻi 5C).
Nā pane ʻano i nā ʻala mimi bipi maoli a me nā mea hana i ka ʻimi ʻana i ka mea hoʻokipa a me ka Anopheles arabianus i hānai ʻia i ke koko. ʻO ke kiʻikuhi o ka olfactometer paipu aniani (A). Ka huki ʻana o nā mea hoʻokahe volatile headspace mai ka mimi bipi hou a me ka ʻelemakule i ka mea hoʻokipa (B) a me nā makika omo koko (C). E huli i ka hopena tentacle o ka Haku An. Nā mea hoʻokahe Headspace i hoʻokaʻawale ʻia mai ka mimi bipi hou (D), 24-hola (E), 72-hola (F), a me 168-hola (G). Hōʻike nā kaha ʻike antenna electron (EAD) i nā loli voltage i ka pane ʻana i nā hui bioactive i loko o ka headspace i hoʻokuʻu ʻia mai ka chromatograph kinoea a ʻike ʻia e kahi mea ʻike ionization lapalapa (FID). Hōʻike ka pā unahi i ka amplitude pane (mV) versus ka manawa paʻa (s). Hōʻike ʻia nā waiwai a me nā helu hoʻokuʻu (µg h-1) o nā hui biologically active. Hōʻike kahi asterisk hoʻokahi (*) i kahi pane haʻahaʻa haʻahaʻa mau. Hōʻike nā asterisk pālua (**) i nā pane hiki ʻole ke hana hou ʻia. E huli i ka mea hoʻokipa (H) a me ka (I) An.arabiensis omo koko he mau ʻano hoʻopiʻi like ʻole i nā hui synthetic o nā ʻala mimi bipi hou a me ka ʻelemakule. ʻO ka awelika o nā makika i huki ʻia i nā inoa leka like ʻole he ʻokoʻa loa kekahi i kekahi (ANOVA hoʻokahi ala me ka hoʻohana ʻana i ka loiloi post hoc a Tukey; p < 0.05). Hōʻike nā kaola hewa i ka hewa maʻamau o ka unahi.
ʻO ka wahine Ann.arabiensis, 72 hola a me 120 hola ma hope o ka ʻai ʻana i ke koko, i ka wā o ka hānau ʻana, ʻaʻohe makemake i hōʻike ʻia no nā mea hoʻokahe monile headspace mai ka mimi bipi hou a me ka ʻelemakule i hoʻohālikelike ʻia me nā kaohi pentane (χ2 = 3.07, p > 0.05; Faila hou 1: Fig. S1).
No ka wahine Ann.arabiensis, ua ʻike ʻia nā loiloi GC-EAD a me GC-MS i ʻewalu, ʻeono, ʻekolu a me ʻekolu mau pūhui bioactive (Kiʻi 5D-G). ʻOiai ua ʻike ʻia nā ʻokoʻa o ka helu o nā pūhui i hoʻoulu ai i nā pane electrophysiological, aia ka hapa nui o kēia mau pūhui i loko o kēlā me kēia ʻāpana volatile headspace i hōʻiliʻili ʻia mai ka mimi hou a me ka mimi ʻelemakule. No laila, no kēlā me kēia ʻāpana, ʻo nā pūhui wale nō i hana i kahi pane physiological mai nā antennae wahine ma luna o ka paepae i hoʻokomo ʻia i nā loiloi hou aʻe.
Ua piʻi aʻe ka nui o ka hoʻokuʻu ʻana o nā mea olaola i loko o ka hōʻiliʻili headspace mai 29 µg h-1 i loko o ka mimi hou a i 242 µg h-1 i loko o ka mimi i hoʻopaʻa ʻia i 168 hola, ma muli o ka piʻi ʻana o ka p-cresol a me ka m-formaldehyde Phenol a me ka phenol. I ka hoʻohālikelike ʻana, ua emi iho nā helu hoʻokuʻu o nā mea olaola ʻē aʻe, e like me 2-cyclohexen-1-one a me decanal, me ka hoʻonui ʻana i ka makahiki o ka mimi, kahi i pili me ka emi ʻana o ka ikaika hōʻailona (nui) i loko o ka chromatogram (Fig. 5D)-G panel hema) a me nā pane physiological i kēia mau mea olaola (Fig. 5D-G panel ʻākau).
Ma keʻano holoʻokoʻa, ua like ka lakio kūlohelohe o nā hui bioactive i ʻike ʻia i loko o nā mea hoʻokahe volatile o nā headspaces mimi hou a me ka ʻelemakule (Kiʻi 5D–G) a ʻaʻole i ʻike ʻia he mea hoihoi nui i ka ʻimi ʻana i kahi kikowaena (χ2 = 8.15, df = 4, p = 0.083; Kiʻi 5H) a i ʻole nā ​​makika omo koko (χ2 = 4.91, df = 4, p = 0.30; Kiʻi 5I). Eia nō naʻe, ua hōʻike nā hoʻohālikelike pālua post hoc ma waena o nā lāʻau lapaʻau he mea hoihoi nui nā makika e ʻimi nei i ka kikowaena i ka hui synthetic o ka mimi 24-h i hoʻohālikelike ʻia me nā kaohi pentane (p = 0.0086; Kiʻi 5H).
No ka loiloi ʻana i ke kuleana o nā ʻāpana pākahi i loko o nā hui synthetic o ka mimi 24-h-makahiki, ua loiloi ʻia nā hui subtractive ʻeono e kūʻē i nā hui piha i ka hoʻāʻo Y-tube, kahi i wehe ʻia ai nā hui pākahi. No nā makika e ʻimi ana i ka host, ʻo ka unuhi ʻana i nā hui pākahi mai ka hui piha he hopena koʻikoʻi i nā pane ʻano (χ2 = 19.63, df = 6, p = 0.0032; Faila hou 1: Kiʻi S2A), ʻoi aku ka hoihoi o nā hui subtractive a pau ma mua o ka Smaller ma mua o ka hui piha ʻana. I ka hoʻohālikelike ʻana, ʻaʻole i hoʻopilikia ka wehe ʻana o nā hui pākahi mai ka hui synthetic piha i nā pane ʻano o nā makika omo koko (χ2 = 11.38, df = 6, p = 0.077), koe wale nō ka decanal, kahi i hopena ai i nā pae haʻahaʻa i hoʻohālikelike ʻia me ka hui piha Attraction (p = 0.022; Faila Hou 1: Kiʻi S2B).
Ma ke kauhale maʻi malaria ma ʻAitiopa, ua loiloi ʻia ka pono o ka hui ʻana o ka mimi pipi 24-hola i ka huki ʻana i nā makika ma lalo o nā kūlana kahua no nā pō he ʻumi (Kiʻi 6A). Ua hopu ʻia a ʻike ʻia he 4,861 mau makika, ʻo 45.7% o ia mau makika he Anthropus.gambiae sl, 18.9% he Anopheles pharoensis a he 35.4% he Culex spp. (Faila hou 1: Papa S1). ʻO Anopheles arabinis wale nō ka lālā o ka paʻakikī ʻano An.Gambian i ʻike ʻia e ka loiloi PCR. Ma ka awelika, ua hopu ʻia he 320 mau makika i kēlā me kēia pō, i loko o ia manawa ua hopu nā pahele me nā maunu hui synthetic i nā makika he nui aʻe ma mua o nā pahele i hui pū ʻia me ka ʻole o ka hui ʻana (χ2(0, 3196) = 170.0, p < 0.0001). Ua hoʻonohonoho ʻia nā pahele ʻaʻole i hoʻokomo ʻia ma kēlā me kēia o nā pō hoʻomalu ʻelima i ka hoʻomaka, waena, a me ka hopena o ka hoʻokolohua. Ua hopu ʻia nā helu like o nā makika i kēlā me kēia pālua o nā pahele, e hōʻike ana ʻaʻohe manaʻo pāʻokoʻa ma waena o nā hale (χ2(0, 1665) = 9 × 10-13, p > 0.05) a ʻaʻohe emi ʻana o ka heluna kanaka i ka wā o ke aʻo ʻana. Ke hoʻohālikelike ʻia me nā pahele kaohi, ua hoʻonui nui ʻia ka nui o nā makika i hopu ʻia i loko o nā pahele i loaʻa ka hui synthetic: ka ʻimi ʻana i ka host (χ2(0, 2107) = 138.7, p < 0.0001), ka hānai koko hou (χ2(0, 650) = 32.2, p < 0.0001) a me ka hāpai ʻana (χ2(0, 228) = 6.27, p = 0.0123; Waihona hou 1: Papa S1). Hōʻike ʻia kēia ma ka huina o nā makika i hopu ʻia: ka ʻimi ʻana i ka host > ka omo koko > ka hāpai > ka hapa hāpai > ke kāne.
Loiloi kahua o ka pono o kahi hui ʻala mimi bipi synthetic 24-hola. Ua mālama ʻia nā hoʻokolohua kahua ma ka hema-waena o ʻAitiopa (palapala ʻāina), kokoke i ke kūlanakauhale ʻo Maki (hoʻokomo), me ka hoʻohana ʻana i kahi pahele kukui Centers for Disease Control (CDC) (ʻākau) ma nā hale i hui pū ʻia, me kahi hoʻolālā huinaha Latin (kiʻi lewa) (A). Hoʻowalewale a hopu nā phototraps CDC i hoʻohui ʻia i ka ʻala synthetic i nā wahine Anopheles arabesques (B), akā ʻaʻole ʻo Anopheles farroes (C), ma ke ʻano ʻokoʻa, kahi hopena physiological-dependent state. Eia kekahi, ua hopu kēia mau pahele i ka nui o nā makika Culex host. (D) Hoʻohālikelike ʻia me ka mana. Hōʻike nā kaola ma ka hema i ka awelika koho ʻana o nā makika i hopu ʻia i loko o nā hui o nā pahele maunu ʻala (ʻōmaʻomaʻo) a me ka mana (hāmama) (N = 10), ʻoiai ʻo nā kaola ma ka ʻākau e hōʻike ana i ka awelika koho ʻana i nā hui o nā pahele mana (hāmama; N = 5). Hōʻike nā Asterisks 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 synthetic. Ke ʻimi nei i ka host (χ2(1, 1345) = 71.7, p < 0.0001), ka hānai koko (χ2(1, 517) = 16.7, p < 0.0001) a me ka hāpai ʻana (χ2(1, 180) = 6.11, p = 0.0134) ua paʻa ʻia ʻo .arabiensis i loko o ka pahele e hoʻokuʻu ana i ka hui synthetic (Kiʻi 6B), ʻoiai ʻaʻole ʻokoʻa ka nui o An. Ua loaʻa ʻo Pharoensis ma nā kūlana physiological like ʻole (Kiʻi 6C). No Culex, ua loaʻa wale ka piʻi nui ʻana o ka nui o nā makika e ʻimi nei i nā host i loko o nā pahele i hoʻokomo ʻia me ka hui synthetic (χ2(1,1319) = 12.6, p = 0.0004; Kiʻi 6D), i hoʻohālikelike ʻia me nā pahele kaohi.
Ua hoʻohana ʻia nā pahele maunu kikowaena i waiho ʻia ma waho o nā kikowaena kūpono ma waena o nā wahi hoʻoulu a me nā kaiāulu kuaʻāina ma ʻAitiopa e loiloi inā hoʻohana nā makika malaria i ka ʻala mimi bipi ma ke ʻano he hōʻailona noho kikowaena. I ka loaʻa ʻole o nā hōʻailona kikowaena, ka wela, a me ka ʻole o ke alo o ka ʻala mimi bipi, ʻaʻohe makika i hopu ʻia (Faila Hou 1: Kiʻi S3). Eia naʻe, i ke alo o ke kiʻekiʻe o ka mahana a me ke ʻala mimi bipi, ua huki ʻia a hopu ʻia nā makika malaria wahine, ʻoiai he liʻiliʻi nā helu, kūʻokoʻa i ka makahiki mimi (χ2(5, 25) = 2.29, p = 0.13; Faila Hou 1: Kiʻi S3). I ka hoʻohālikelike ʻana, ʻaʻole i hopu nā kaohi wai i nā makika malaria i nā mahana kiʻekiʻe (Faila Hou 1: Kiʻi S3).
Loaʻa a hoʻolaha nā makika malaria i nā hui nitrogen ma o ka hānai uku ʻana i ka mimi bipi (ʻo ia hoʻi, nā puddles) e hoʻomaikaʻi i nā ʻano mōʻaukala ola, e like me nā pepeke ʻē aʻe [2, 4, 24, 25, 26]. ʻO ka mimi bipi kahi kumuwaiwai hou i loaʻa koke e pili kokoke ana i nā wahi hoʻomaha no nā mea lawe malaria, e like me nā hale bipi a me nā mea kanu kiʻekiʻe kokoke i nā hale kuaʻāina a me nā wahi hānau. Loaʻa i nā makika wahine kēia kumuwaiwai ma o ka honi a hiki ke hoʻoponopono i ka lawe ʻana o nā hui nitrogenous i loko o ka mimi, me ka urea, ka ʻāpana nitrogenous nui i loko o ka mimi [15, 16]. Ma muli o ke kūlana physiological o ka makika wahine, ua hoʻokaʻawale ʻia nā meaʻai i loko o ka mimi e hoʻomaikaʻi i ka hana lele a me ke ola ʻana o nā makika wahine e ʻimi nei i ka host, a me ke ola a me nā ʻano hānau o nā kānaka i hānai ʻia i ke koko i ka wā o ka pōʻaiapuni gonadotropic mua. No laila, he kuleana koʻikoʻi ka hui ʻana o ka mimi no nā mea lawe malaria i pani ʻia e like me nā pākeke malnutried [8], no ka mea e hāʻawi ana i nā makika wahine me ka hiki ke loaʻa nā hui nitrogenous koʻikoʻi ma ke komo ʻana i ka hānai haʻahaʻa. Loaʻa i kēia ʻike nā hopena epidemiological koʻikoʻi, ʻoiai e hoʻonui ana nā wahine i ko lākou ola, ka hana a me ka hopena hānau hou, nā mea āpau e hoʻopilikia i ka hiki ke vector. Eia kekahi, ʻo kēia ʻano hana paha ke pahuhopu o nā papahana hoʻokele vector e hiki mai ana.