Samun abinci mai gina jiki da kuma rarrabawa yana haɗa halayen neman kwari da tarihin rayuwa. Domin rama ƙarancin wasu sinadarai masu gina jiki a matakai daban-daban na rayuwa, kwari za su iya samun waɗannan sinadarai ta hanyar ciyarwa ta ƙarin abinci, misali, ta hanyar ciyar da ƙwayoyin halitta masu ƙashi a cikin wani tsari da aka sani da kududdufi. Sauro Anopheles arabiani ya bayyana yana da rashin abinci mai gina jiki, saboda haka, yana buƙatar sinadarai masu gina jiki don metabolism da haifuwa. Manufar wannan binciken ita ce tantance ko tashin hankalin An. arabiensis akan fitsarin saniya don samun sinadarai masu gina jiki yana inganta halayen tarihin rayuwa.
Tabbatar cewa yana da aminci. arabiensis ya sami sha'awar ƙamshin fitsarin saniya sabo, na awanni 24, na awanni 72, da na awanni 168, kuma an auna mata masu neman haihuwa da kuma waɗanda aka shayar da jini (sa'o'i 48 bayan cin abinci) a cikin na'urar auna bututun Y, kuma an tantance mata masu juna biyu don gwajin haihuwa. Daga nan aka yi amfani da haɗin sinadarai da na lantarki don gano mahaɗan halitta a cikin fitsarin saniya a dukkan azuzuwan shekaru huɗu. An kimanta gaurayen mahaɗan halitta na halitta a cikin gwaje-gwajen bututun Y da filin. Don bincika fitsarin saniya da babban mahaɗan nitrogen ɗinsa a matsayin abincin da zai iya zama ƙarin abinci ga masu cutar malaria, an auna sigogin ciyarwa da halayen tarihin rayuwa. An tantance adadin sauro mata da adadin fitsarin saniya da urea da aka sha. Bayan ciyarwa, an tantance mata don rayuwa, tashi da haihuwa, da kuma haihuwa.
Nemi jinin mai masaukin baki da abinci mai gina jiki. A cikin binciken dakin gwaje-gwaje da na filin, Larabawa sun sha'awar ƙamshin halitta da na roba na fitsarin saniya sabo da tsufa. Mata masu juna biyu ba sa damuwa da amsawar fitsarin saniya a wuraren haihuwa. Mata masu neman mai masaukin baki da tsotsar jini suna shan fitsarin saniya da urea kuma suna ware waɗannan albarkatu bisa ga canjin tarihin rayuwa a matsayin aikin yanayin jiki don tashi, rayuwa, ko haihuwa.
Samun fitsarin saniya da rarraba shi a cikin Anopheles arabinis don inganta halayen tarihin rayuwa. Karin ciyar da fitsarin saniya yana shafar ƙarfin vector kai tsaye ta hanyar ƙara yawan rayuwa ta yau da kullun da yawan vector, kuma a kaikaice ta hanyar canza ayyukan tashi, don haka ya kamata a yi la'akari da shi a cikin samfuran gaba.
Sayen abinci mai gina jiki da rarrabawa yana haɗa hanyoyin neman kwari da halayen tarihin rayuwa [1,2,3]. Kwari suna iya zaɓar abinci da kuma samunsa da kuma yin diyya bisa ga wadatar abinci da buƙatun gina jiki [1, 3]. Rarraba abinci mai gina jiki ya dogara ne akan tsarin rayuwa da tarihin rayuwa kuma yana iya haifar da buƙatu daban-daban don ingancin abinci da yawa a matakai daban-daban na rayuwar kwari [1, 2]. Don rama ƙarancin abinci mai gina jiki na musamman, kwari na iya samun waɗannan abubuwan gina jiki ta hanyar ciyarwa ta ƙarin abinci, kamar a kan laka, najasa daban-daban da kuma fitar da ƙwayoyin halitta masu ƙashi, da kuma mushe, wani tsari da aka sani da kududdufai [2]. Kodayake an bayyana nau'ikan nau'ikan malam buɗe ido da ƙwari iri-iri, ramukan ban ruwa suma suna faruwa a wasu tsari na kwari, kuma jan hankali da ciyar da waɗannan nau'ikan albarkatu na iya yin tasiri mai mahimmanci akan lafiya da sauran halaye na tarihin rayuwa [2, 4, 5, 6], 7]. Sauro na zazzabin cizon sauro Anopheles gambiae sensu lato (sl) ya fito a matsayin babba 'marasa abinci mai gina jiki' [8], don haka shayarwa na iya taka muhimmiyar rawa a cikin halayen tarihin rayuwarsa, amma wannan ɗabi'ar tana da zuwa yanzu an yi watsi da su. Amfani da tashin hankali a matsayin hanyar ƙara yawan sinadarin gina jiki a cikin wannan muhimmin abin hawa yana buƙatar kulawa domin wannan na iya haifar da sakamako masu mahimmanci na annoba.
Shan sinadarin Nitrogen a cikin sauro na Anopheles na mata manya yana da iyaka saboda ƙarancin adadin kuzari da ake samu daga matakin tsutsa da rashin amfani da abincin jini yadda ya kamata [9]. Mace Ann.gambiae sl yawanci tana rama wannan ta hanyar ƙara musu abinci na ƙarin jini [10, 11], wanda hakan ke sanya mutane da yawa cikin haɗarin kamuwa da cutar da kuma sanya sauro cikin haɗarin farauta. A madadin haka, sauro na iya amfani da ƙarin ciyar da ƙashi na ƙashi don samun mahaɗan nitrogen waɗanda ke haɓaka daidaitawa da iya motsawa, kamar yadda wasu kwari suka nuna [2]. A wannan batun, jan hankalin ɗaya daga cikin nau'in 'yan'uwa a cikin An. Tsarin nau'in sl na Gambia, Anopheles arabinis, sabon fitsarin saniya da tsufa [12,13,14], abin sha'awa ne. Anopheles arabinis yana da damar yin amfani da damar da yake da ita kuma an san yana da alaƙa da kuma ciyar da shanu. Fitsari na saniya albarkatu ne masu wadataccen sinadarai na nitrogen, tare da urea wanda ke da kashi 50-95% na jimlar nitrogen a cikin fitsari sabo [15, 16]. Kamar yadda fitsarin saniya yake Shekaru masu yawa, ƙananan halittu suna amfani da waɗannan albarkatun don rage sarkakiyar mahaɗan nitrogen cikin awanni 24 [15]. Tare da ƙaruwar ammonia cikin sauri, wanda ke da alaƙa da raguwar nitrogen na halitta, ƙananan halittun alkaloid (da yawa daga cikinsu suna samar da mahaɗan da ke da guba ga sauro) suna bunƙasa [15], wanda wataƙila mace ce Ann.arabiensis tana sha'awar fitsari bayan sa'o'i 24 ko ƙasa da haka [13, 14].
A cikin wannan binciken, an nemi Ans masu masaukin baki da waɗanda aka shayar da jini. A lokacin zagayen gonadotropin na farko, an tantance arabiensis don samun mahaɗan nitrogen, gami da urea, ta hanyar haɗa fitsari. Na gaba, an gudanar da jerin gwaje-gwaje don tantance yadda sauro mata ke ware wannan albarkatun gina jiki mai yuwuwa don inganta rayuwa, haifuwa da kuma ƙarin neman abinci. A ƙarshe, an tantance warin fitsarin saniya sabo da tsufa don tantance ko waɗannan sun ba da alamu masu inganci ga An mai masaukin baki da wanda aka shayar da jini. A cikin bincikensu na wannan albarkatun abinci mai yuwuwa, arabiensis ya gano alaƙar sinadarai a bayan bambancin kyawun da aka lura. An ƙara kimanta gaurayawan warin roba na mahaɗan halitta masu canzawa (VOCs) waɗanda aka gano a cikin fitsari mai shekaru 24 a ƙarƙashin yanayin filin, suna faɗaɗa sakamakon da aka samu a ƙarƙashin yanayin dakin gwaje-gwaje da kuma nuna tasirin warin fitsarin shanu akan yanayin ilimin halittar jiki daban-daban. Jan hankalin sauro. Sakamakon da aka samu ya tabbatar da cewa An. arabiensis yana samun kuma yana rarraba mahaɗan nitrogen da aka samu a cikin fitsarin ƙashi don yin tasiri ga halayen tarihin rayuwa. An tattauna waɗannan sakamakon a cikin mahallin tasirin cututtuka da kuma yadda za a iya amfani da su don sa ido da sarrafa vector.
An kiyaye nau'in Anopheles arabic (irin Dongola) a zafin 25 ± 2 °C, 65 ± 5% RH da kuma zagayowar haske: duhu na awanni 12:12. An kiwon tsutsotsi a cikin tiren filastik (20 cm × 18 cm × 7 cm) cike da ruwa mai narkewa sannan aka ciyar da abincin kifi na Tetramin® (Tetra Werke, Melle, DE). An tattara tsutsotsi a cikin kofuna 30 ml (Nolato Hertila, Åstorp, SE) sannan aka mayar da su zuwa kejin Bugdorm (30 cm × 30 cm × 30 cm; MegaView Science, Taichung, Taiwan) don ba da damar girma. An ba wa manya maganin sucrose 10% ad libitum har zuwa kwanaki 4 bayan fitowar (dpe), a lokacin da aka bai wa mata masu neman masauki abinci nan da nan kafin gwajin, ko kuma aka bar su cikin yunwa da ruwa mai narkewa kafin gwajin, kamar yadda aka bayyana a ƙasa. An bai wa mata da aka yi amfani da su don gwaje-gwajen bututun jirgin sama abinci na tsawon awanni 4-6 kawai. awanni da ruwa da libitum. Domin shirya sauro masu tsotsar jini don gwaje-gwajen bio na gaba, an ba wa mata 4 dpe jinin tumaki masu defibrotic (Håtunalab, Bro, SE) ta amfani da tsarin ciyar da membrane (Hemotek Discovery Workshops, Accrington, UK). Daga nan aka mayar da mata masu cike da cunkoso zuwa keji daban-daban kuma aka ba su abinci kai tsaye, kamar yadda aka bayyana a ƙasa, ko kuma 10% sucrose ad libitum na tsawon kwanaki 3 kafin gwaje-gwajen da aka bayyana a ƙasa. An yi amfani da mata na ƙarshe don gwajin bioasay na bututun jirgin sama kuma aka mayar da su zuwa dakin gwaje-gwaje, sannan aka yi musu distilled ruwa ad libitum na tsawon awanni 4-6 kafin gwajin.
An yi amfani da gwajin ciyarwa don auna yawan shan fitsari da urea ga mace 'yar An. Arab. An ba wa mata masu neman mai masaukin baki da masu shan jini abinci mai ɗauke da kashi 1% na fitsarin saniya sabo da tsufa, yawan urea daban-daban, da kuma sarrafawa guda biyu (10% sucrose da ruwa) na tsawon awanni 48. Bugu da ƙari, an ƙara launin abinci (1 mg ml-1 xylene cyanide FF; CAS 2650-17-1; Sigma-Aldrich, Stockholm, SE) a cikin abincin kuma an samar da shi a cikin matrix 4 × 4 a cikin bututun microcentrifuge 250 µl (Axygen Scientific, Union City, CA, US; Hoto na 1A) Cika har zuwa gefen (~300 µl). Don guje wa gasa tsakanin sauro da yuwuwar tasirin launin rini, sanya sauro 10 a cikin babban kwano na Petri (diamita 12 cm da tsayi 6 cm; Semadeni, Ostermundigen, CH; Hoto na 1A) a cikin duhu gaba ɗaya a 25 ± 2 cm °C da 65 ± 5% dangi Danshi. An maimaita waɗannan gwaje-gwajen sau 5 zuwa 10. Bayan an sha wahala wajen cin abinci, an sanya sauro a -20 °C har sai an ci gaba da bincike.
Nemi fitsarin shanu da urea da mace mai masaukin baki da tsotsar jini Anopheles arabianus ta sha. A cikin gwajin ciyarwa (A), an ba wa sauro mata abinci mai gina jiki wanda ya ƙunshi fitsarin saniya sabo da tsufa, yawan urea daban-daban, sucrose (10%), da ruwan da aka tace (H2O). Mata masu neman mai masaukin baki (B) da waɗanda aka ciyar da jini (C) sun sha ƙarin sucrose fiye da kowace abinci da aka gwada. Lura cewa mata masu neman mai masaukin baki suna shan fitsarin saniya na awanni 72 ƙasa da fitsarin saniya na awanni 168 (B). Matsakaicin adadin nitrogen (± karkacewar daidaito) na fitsari an wakilta shi a cikin akwatin. Mata masu neman mai masaukin baki (D, F) da tsotsar jini (E, G) suna shan urea ta hanyar da ta dogara da kashi. Matsakaicin girman da aka shaƙa (D, E) tare da sunayen haruffa daban-daban sun bambanta sosai da juna (ANOVA ta hanya ɗaya ta amfani da nazarin Tukey bayan hoc; p < 0.05). Sandunan kuskure suna wakiltar kuskuren daidaito na matsakaicin (BE). Layin madaidaiciya yana wakiltar layin komawar layi (F, G)
Domin sakin abincin da ya sha, an sanya sauro daban-daban a cikin bututun microcentrifuge na 1.5 ml wanda ke ɗauke da 230 µl na ruwan da aka tace sannan aka lalata kyallen ta amfani da injin da aka zubar da shi da kuma injin mara waya (VWR International, Lund, SE), sannan aka yi amfani da centrifugation a 10 krpm na tsawon minti 10. An mayar da ruwan sama (200 µl) zuwa wani rami mai rami 96 (Sigma-Aldrich) kuma an tantance sha (λ620) ta amfani da na'urar karanta microplate mai tushen spectrophotometer (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm). A madadin haka, an niƙa sauro a cikin 1 ml na ruwan da aka tace, 900 µl daga ciki an canja shi zuwa cuvette don nazarin spectrophotometric (λ 620 nm; UV 1800, Shimadzu, Kista, SE). Don ƙididdige yawan abincin da ake ci, an shirya madaidaicin lanƙwasa ta hanyar dilution na serial don samar da 0.2 µl. zuwa 2.4 µl na 1 mg ml-1 xylene cyanide. Sannan, an yi amfani da yawan gani na yawan rini da aka sani don tantance adadin abincin da kowace sauro ta ci.
An yi nazarin bayanai na girma ta amfani da nazarin bambancin hanya ɗaya (ANOVA) sannan aka yi kwatancen Tukey bayan hoc biyu (JMP Pro, v14.0.0, SAS Institute Inc., Cary, NC, US, 1989–2007). Binciken juye-juye ya bayyana yawan shan urea da ya dogara da yawan mai da hankali da kuma kwatanta martani tsakanin neman mai masaukin baki da kuma shan jini (GraphPad Prism v8.0.0 don Mac, GraphPad Software, San Diego, CA, Amurka).
An ɗaure kimanin µl na samfuran fitsari daga kowace rukuni na shekaru a kan Chromosorb® W/AW (10 mg 80/100 mesh, Sigma Aldrich) kuma an lulluɓe su a cikin kwalin tin (8 mm × 5 mm). An saka kwalin a cikin ɗakin konewa na na'urar nazarin CHNS/O (Flash 2000, Thermo Fisher Scientific, Waltham, MA, US) don tantance yawan nitrogen a cikin fitsari sabo da tsufa bisa ga ka'idar masana'anta. An auna jimlar nitrogen (g N l-1) bisa ga yawan urea da aka sani da aka yi amfani da shi azaman ma'auni.
Domin tantance tasirin abinci ga rayuwar mata masu neman mai masaukin baki da tsotsar jini, an sanya sauro daban-daban a cikin manyan kwano na Petri (diamita 12 cm da tsayi 6 cm; Semadeni) tare da rami mai rufe raga a cikin murfi (diamita 3 cm) tare da Don samun iska da wadatar abinci. An samar da abinci kai tsaye bayan 4 dpe kuma ya haɗa da 1% na fitsarin saniya sabo da tsufa, yawan urea guda huɗu, da kuma sarrafawa guda biyu, 10% sucrose da ruwa. An saka kowace abinci a kan tampon na hakori (DAB Dental AB, Upplands Väsby, SE) a cikin sirinji na 5 ml (Thermo Fisher Scientific, Gothenburg, SE), an cire bututun, aka sanya a saman kwano na petri (siffa ta 1).1A). Canza abincin ku kowace rana. Kula da dakin gwaje-gwaje kamar yadda aka bayyana a sama. An ƙidaya sauro da suka tsira sau biyu a rana, yayin da aka jefar da sauro da suka mutu har sai sauro na ƙarshe ya mutu (n = 40 a kowace magani). An yi nazarin rayuwar sauro da aka ciyar da su a kan abinci daban-daban ta amfani da lanƙwasa na rayuwa na Kaplan-Meyer da matsayi na log-ranking. gwaje-gwaje don kwatanta kwatancen rarraba rayuwa tsakanin abinci (IBM SPSS Statistics 24.0.0.0).
Injin niƙa na musamman na sauro wanda aka gina bisa ga Attisano da abokan aikinsa.[17], wanda aka yi da bangarorin acrylic masu kauri mm 5 (faɗi cm 10 x tsayi cm 10 x tsayi cm 10) ba tare da bangarorin gaba da baya ba (Hoto na 3: sama). Haɗaɗɗen bututu mai tsayi wanda aka yi da ginshiƙin gas chromatography (0.25 mm id; 7.5 cm L) tare da ƙarshen da aka manne a kan allurar kwari da aka rataye tsakanin maganadisu na neodymium guda biyu a nesa da cm 9. Bututun kwance da aka yi da abu ɗaya (6.5 cm L) ya raba bututun tsaye don samar da hannu da aka ɗaure da hannu wanda ke ɗauke da ƙaramin foil na aluminum a matsayin siginar katse haske.
An bai wa mata masu sa'o'i 24 abincin da ke sama na tsawon mintuna 30 kafin a dakatar da su. Sannan aka yi wa sauro mata da aka ciyar da su cikakken magani a kan kankara na tsawon mintuna 2-3 sannan aka haɗa su da kakin zuma (Joel Svenssons Vaxfabrik AB, Munka Ljungby, SE) sannan aka ɗaure su da hannun bututun kwance. Na'urar Tashi. An yi rikodin juyin juya hali a kowace tashi ta hanyar mai tattara bayanai na musamman, sannan aka adana su kuma aka nuna su ta amfani da software na PC-Lab 2000™ (v4.01; Velleman, Gavere, BE). An sanya injin niƙa jirgin a cikin ɗaki da aka tsara yanayi (awanni 12:12, haske: duhu, 25 ± 2 °C, 65 ± 5% RH).
Domin hango yanayin aikin jirgin, an ƙididdige jimlar nisan da aka yi tafiya (m) da jimlar adadin ayyukan jirgin a jere a kowace awa a cikin tsawon awanni 24. Bugu da ƙari, an kwatanta matsakaicin nisan da aka yi tafiya da mata a cikin jiyya kuma an yi nazari ta amfani da ANOVA ta hanya ɗaya da kuma nazarin Tukey bayan hoc (JMP Pro, v14.0.0, SAS Institute Inc.), inda aka ɗauki matsakaicin nisan a matsayin mai dogaro, yayin da magani abu ne mai zaman kansa. Bugu da ƙari, ana ƙididdige matsakaicin adadin zagaye a cikin ƙarin mintuna 10.
Domin tantance tasirin abinci akan aikin haihuwa na An.arabiensis, an mayar da mata shida (4 dpe) kai tsaye zuwa kejin Bugdorm (30 cm × 30 cm × 30 cm) bayan an tattara jini sannan aka ba da abincin gwaji na tsawon awanni 48 kamar yadda aka bayyana a sama. Daga nan aka cire abincin sannan aka ba da kofunan haihuwa (30 ml; Nolato Hertila) cike da ruwa mai narkewa 20 ml a rana ta uku na tsawon awanni 48, ana canza su duk bayan awanni 24. A maimaita kowace tsarin abinci sau 20-50. An ƙidaya ƙwai kuma an yi rikodin su ga kowane keji na gwaji. An yi amfani da ƙananan samfuran ƙwai don tantance matsakaicin girma da bambancin tsayi na ƙwai ɗaya (n ≥ 200 a kowace abinci) ta amfani da na'urar hangen nesa ta Dialux-20 (DM1000; Ernst Leitz Wetzlar, Wetzlar, DE) wacce aka sanye da kyamarar Leica (DFC) 320 R2; Leica Microsystems Ltd., DE). An ajiye sauran ƙwai a cikin ɗakin da ke da yanayin yanayi a ƙarƙashin yanayin kiwo na yau da kullun na tsawon awanni 24, kuma an auna ƙaramin samfurin tsutsotsi na farko da suka fito kwanan nan (n ≥ 200 a kowace abinci), kamar yadda aka bayyana a sama. An kwatanta adadin ƙwai da girman ƙwai da tsutsotsi tsakanin jiyya da amfani da ANOVA na hanya ɗaya da kuma nazarin Tukey bayan hoc (JMP Pro, v14.0.0, SAS Institute Inc.).
An tattara fitsarin da aka samo daga sabo (awa 1 bayan an yi gwajin), fitsarin da ya tsufa na awanni 24, awanni 72 da awanni 168 daga samfuran da aka tattara daga shanun Zebu, Arsi. Don saukaka, an tattara samfuran fitsari da sassafe yayin da shanun ke cikin rumbun ajiya. An tattara samfuran fitsari daga mutane 10 kuma an canja 100-200 ml na kowane samfurin zuwa jakunkunan yin burodi na polyamide daban-daban (Toppits Cofresco, Frischhalteprodukte GmbH da Co., Minden, DE) a cikin ganga na filastik na vinyl chloride lita 3. An tattara ƙwayoyin cuta daga kowane samfurin fitsari na shanu ko dai kai tsaye (sabo) ko kuma bayan sun girma a zafin ɗaki na awanni 24, awanni 72 da awanni 168, watau kowane samfurin fitsari yana wakiltar kowace rukunin shekaru.
Don tattara ƙwayoyin cuta masu rai a kan kai, an yi amfani da tsarin rufewa don yaɗa kwararar iskar gas mai tace carbon (100 ml min-1) ta cikin jakar polyamide zuwa ginshiƙin shaƙatawa na tsawon awanni 2.5 ta amfani da famfon tsotsar iskar diaphragm (KNF Neuberger, Freiburg, DE). A matsayin mai sarrafawa, an yi tattara sararin sama daga jakar polyamide mara komai. An yi ginshiƙin shaƙatawa da bututun Teflon (5.5 cm x 3 mm id) wanda ke ɗauke da 35 MG na Porapak Q (50/80 mesh; Waters Associates, Milford, MA, US) tsakanin matosai na ulu na gilashi. Kafin amfani, an wanke ginshiƙin da n-hexane mai rai 1 ml (Merck, Darmstadt, DE) da pentane 1 ml (99.0% tsantsar GC grade, Sigma Aldrich). An cire ƙwayoyin cuta masu rai da 400 μl na pentane. An tattara tarin sararin sama sannan a adana su a -20°C har sai an yi amfani da su don ƙarin amfani. bincike.
Amsoshin ɗabi'a na neman mai masauki da kuma cin jini An. An yi nazarin fitar da fitsari mai canzawa daga saman jiki da aka tattara daga sabo, awanni 24, awanni 72, da awanni 168 don gano fitar da iska mai canzawa daga sauro na Arabidopsis ta amfani da bututun gilashi madaidaiciya olfactometer [18]. An gudanar da gwaje-gwajen a lokacin ZT 13-15, lokacin mafi girman aikin neman gida na An. Arab [19]. An haskaka wani bututun gilashi olfactometer (80 cm × 9.5 cm id) da haske ja 3 ± 1 lx daga sama. Iska mai tacewa da danshi ta gawayi (25 ± 2 °C, 65 ± 2% danshin dangi) ta wuce gwajin bioasay a 30 cm s-1. An ratsa iska ta cikin jerin allon raga na bakin karfe, yana samar da kwararar laminar da tsarin plum iri ɗaya. Na'urar rarraba tampon na haƙori (4 cm × 1 cm; L:D; DAB Dental AB), an dakatar da shi daga na'urar 5 cm a ƙarshen iska na'urar auna olfactometer, tare da canza mai motsa jiki a kowane minti 5. Don yin nazari, an yi amfani da 10 μl na kowane cirewar sararin kai, wanda aka narkar da shi 1:10, a matsayin abin ƙarfafawa. An yi amfani da daidai adadin pentane a matsayin abin sarrafawa. An sanya sauro masu neman mai masauki ko tsotsar jini a cikin kejin sakin mutum ɗaya awanni 2-3 kafin fara gwajin. An sanya kejin sakin a gefen iska na olfactometer, kuma an bar sauro su daidaita na minti 1, sannan aka buɗe bawul ɗin malam buɗe ido na keji don a sake shi. An bincika jan hankalin magani ko iko a matsayin rabon sauro da suka taɓa tushen a cikin mintuna 5 bayan an sake shi. An kwafi kowane cirewar sararin kai da sarrafawa mai canzawa aƙalla sau 30, kuma don guje wa tasirin kowace rana guda, an gwada adadin magunguna da sarrafawa iri ɗaya a kowace rana ta gwaji. Nemi amsoshi daga mai masauki da mai shan jini Ans. An bincika saitin sararin kai da na kai ta amfani da koma-baya na ƙididdiga na asali sannan aka biyo baya da kwatancen biyu don rabo mara kyau (JMP Pro, v14.0.0, SAS Institute Inc.).
Amsar haihuwar An. An yi nazarin abubuwan da aka samo daga fitsarin saniya sabo da tsufa a cikin kejin Bugdorm (30 cm × 30 cm × 30 cm; MegaView Science). Kofuna na filastik (30 mL; Nolato Hertila) da aka cika da ruwa mai narkewa 20 mL sun samar da tushen haihuwa kuma an sanya su a kusurwoyi daban-daban na kejin, 24 cm a tsakaninsu. An daidaita kofunan magani da 10 μl na kowane cirewar kai a cikin ruwa mai narkewa 1:10. An yi amfani da adadin pentane daidai don daidaita kofin sarrafawa. An yi musayar kofunan magani da sarrafawa tsakanin kowace gwaji don sarrafa tasirin matsayi. An saki mata goma da aka shayar da jini a cikin kejin gwaji a ZT 9-11 kuma an ƙidaya ƙwai a cikin kofuna bayan sa'o'i 24. Tsarin lissafin ma'aunin haihuwar shine: (adadin ƙwai da aka saka a cikin kofin magani - adadin ƙwai da aka saka a cikin kofin sarrafawa)/(jimillar adadin ƙwai da aka saka). An maimaita kowace magani sau 8.
An yi nazarin tsarin gano yanayin eriya ta gas da electron (GC-EAD) na An.arabiensis na mace kamar yadda aka bayyana a baya [20]. A takaice, an raba sabbin abubuwan da ke canza yanayin kan kai ta amfani da Agilent Technologies 6890 GC (Santa Clara, CA, US) wanda aka sanye da ginshiƙi na HP-5 (30 m × 0.25 mm id, kauri fim na 0.25 μm, Agilent Technologies). da fitsari mai tsufa. An yi amfani da Hydrogen a matsayin matakin motsi tare da matsakaicin gudu mai layi na 45 cm s-1. An yi wa kowane samfurin (2 μl) allura na tsawon daƙiƙa 30 a yanayin da ba a raba shi ba tare da zafin shiga na 225 °C. An tsara zafin tanda na GC daga 35 °C (riƙewa na minti 3) zuwa 300 °C (riƙewa na minti 10) a 10 °C min-1. A cikin mai raba fitar da iska na GC, an ƙara 4 psi na nitrogen kuma an raba 1:1 a cikin Gerstel 3D/2 low dead volume cross (Gerstel, Mülheim, DE) tsakanin na'urar gano ionization na harshen wuta da EAD. An wuce bututun fitar da iska na GC don EAD ta hanyar layin canja wurin Gerstel ODP-2, wanda ke bin diddigin zafin tanda na GC da 5 °C, zuwa cikin bututun gilashi (10 cm × 8 mm), inda aka haɗa shi da iska mai tace carbon, mai danshi (1.5 l min−1). An sanya eriya ɗin. 0.5 cm daga maɓuɓɓugar bututun. Kowace sauro ta yi kwafi ɗaya, kuma ga sauro masu neman masauki, an yi aƙalla kwafi uku a kan samfuran fitsari na kowane zamani.
Gano mahaɗan halittu masu aiki a cikin tarin sararin samaniya na fitsarin shanu sabo da tsufa ta amfani da na'urar auna GC da mass spectrometer (GC-MS; 6890 GC da 5975 MS; Agilent Technologies) don samar da martanin eriya a cikin nazarin GC-EAD, yana aiki a yanayin tasirin ionization na electron a 70 eV. An sanye GC da ginshiƙin silica mai rufi da aka haɗa da HP-5MS UI (diamita na ciki 60 m × 0.25 mm, kauri fim 0.25 μm) ta amfani da helium a matsayin matakin motsi tare da matsakaicin kwararar layi na 35 cm s-1. An allurar samfurin 2 μl ta amfani da saitunan allura iri ɗaya da zafin tanda kamar yadda aka yi da nazarin GC-EAD. An gano mahaɗan dangane da lokacin riƙe su (Kovát index) da kuma ma'aunin taro idan aka kwatanta da ɗakin karatu na musamman da ɗakin karatu na NIST14 (Agilent). An tabbatar da mahaɗan da aka gano ta hanyar allurar ingantattun ƙa'idodi (Ƙarin Fayil na 1: Tebur S2). Don ƙididdigewa, heptyl acetate An yi allurar allurar a matsayin mizani na waje (10 ng, 99.8% tsarkin sinadarai, Aldrich).
Kimanta ingancin cakuda wari na roba wanda ya ƙunshi mahaɗan halitta masu aiki da aka gano a cikin fitsari sabo da tsufa don jawo hankalin masu neman mai gida da tsotsar jini Ans.arabiensis, ta amfani da ma'aunin olfactomer da yarjejeniya iri ɗaya kamar yadda aka ambata a sama. Haɗaɗɗun roba sun kwaikwayi abun da ke ciki da rabon mahaɗan a cikin gaurayen sararin sama na fitsari sabo, awanni 24, awanni 48, awanni 72, da awanni 168 da suka tsufa (Hoto na 5D-G; Ƙarin Fayil na 1: Tebur S2). Don bincike, yi amfani da 10 μl na 1:100 na cakuda cikakke, tare da jimlar fitarwa daga kimanin 140-2400 ng h-1, don tantance kyawun sauro mai masauki da tsotsar jini. Bayan haka, ana yin gwajin akan gauraye cikakke, inda ake cire gauraye masu ragewa na mahaɗan guda ɗaya na cikakken cakuda. Nemi amsoshi daga mai masauki da mai shan jini Ans.Arab vs gauraye masu roba da raguwa ta amfani da koma-baya na ƙididdiga na asali sannan kuma kwatantawa biyu don abubuwan da ba a saba gani ba rabo (JMP Pro, v14.0.0, SAS Institute Inc.).
Domin tantance ko fitsarin saniya zai iya zama alamar wurin zama ga sauro na maleriya, fitsarin saniya sabo da tsohuwa, wanda aka tattara kamar yadda aka bayyana a sama, sannan aka sanya ruwa a cikin bokiti mai lita 3 (100 ml) sannan aka sanya shi a cikin tarkon kiwo na mai masaukin baki. (Sigar BG-HDT; BioGents, Regensburg, DE). Tarkuna goma an sanya su a nisan mita 50 a cikin kiwo, mita 400 daga al'ummar ƙauye (Silay, Habasha, 5°53´24´´N, 37°29´24´´E) kuma babu shanu, a wuraren kiwo na dindindin da ƙauyuka. An dumama tarkuna biyar don kwaikwayon kasancewar mai masaukin baki, yayin da aka bar tarkuna biyar ba tare da dumama ba. Kowace wurin magani ana juyawa kowace dare tsawon dare biyar. An kwatanta adadin sauro da aka kama a cikin tarkunan da aka kama da fitsari na shekaru daban-daban ta amfani da tsarin logistic regression tare da rarraba beta binomial (JMP Pro, v14.0.0, SAS Institute Inc.).
A wani ƙauye da ke fama da cutar maleriya kusa da garin Maki, yankin Oromia, Habasha (8° 11′ 08″ N, 38° 81′ 70″ E; Hoto na 6A). An gudanar da binciken tsakanin tsakiyar watan Agusta da tsakiyar Satumba kafin feshin da ya rage a cikin gida na shekara-shekara, tare da damina mai tsawo. An zaɓi gidaje guda biyar (mita 20-50 daban) waɗanda ke gefen ƙauyen don binciken (Hoto na 6A). Sharuɗɗan da aka yi amfani da su don zaɓar gidajen sune: ba a yarda da dabbobi a cikin gidan ba, ba a yarda da dafa abinci a cikin gida ba (zanen itace ko gawayi) (aƙalla a lokacin gwaji), da gidaje masu matsakaicin mazauna biyu, suna kwana a cikin ƙwayoyin cuta marasa kwari. a ƙarƙashin gidan sauro da aka yi wa magani. Hukumar Binciken Ɗabi'un Bincike ta Cibiya (IRB/022/2016) ta Kwalejin Kimiyyar Halittu (CNS-IRB), Jami'ar Addis Ababa, ta amince da ɗabi'a bisa ga ka'idojin da aka kafa a sanarwar Helsinki ta Ƙungiyar Likitoci ta Duniya. An sami izini daga kowane shugaban gida tare da taimakon ma'aikatan faɗaɗa lafiya. Hukumomin gida a matakin gunduma da unguwa ('kebele') sun amince da dukkan tsarin. Tsarin gwajin ya biyo bayan ƙirar murabba'in Latin 2 × 2, inda aka sanya gauraye da sarrafawa na roba ga gidaje biyu a daren farko kuma aka canza su tsakanin gidaje a daren gwaji na gaba. An maimaita wannan tsari sau goma. Bugu da ƙari, don kimanta ayyukan sauro a cikin gidaje da aka zaɓa, an saita tarkunan CDC don gudanar da dare biyar a jere a farkon, tsakiya da ƙarshen gwajin filin a lokaci guda na rana.
An narkar da wani cakuda mai gina jiki wanda ke ɗauke da sinadarai shida masu aiki a cikin heptane (97.0% GC mai narkewa, Sigma Aldrich) kuma an sake shi a 140 ng h-1 ta amfani da na'urar rarraba wick na auduga [20]. Na'urar rarraba wick ta ba da damar sakin dukkan sinadarai a cikin daidaito a cikin gwajin sa'o'i 12. An yi amfani da Heptane a matsayin magani. An rataye kwalbar kusa da wurin shiga tarkon haske na Cibiyar Kula da Cututtuka da Rigakafi (CDC) (John W. Hock Company, Gainesville, FL, US; Hoto na 6A). An rataye tarkunan a mita 0.8 - 1 sama da ƙasa, kusa da ƙafar gado, kuma wata mai sa kai ta kwana a ƙarƙashin gidan sauro wanda ba a yi masa magani ba kuma an yi mata aiki tsakanin ƙarfe 18:00 da 06:30. Daga baya an tantance sauro da aka kama ta hanyar jinsi da yanayin jiki (ba a ciyar da su ba, ba a ciyar da su ba, ba su da juna biyu, kuma ba su da juna biyu [21] daga baya an tantance su ta amfani da nazarin polymerase chain reaction (PCR) don gano nau'in da aka gano a matsayin A. gambiae sl. Membobin hadaddun [23]. A cikin binciken filin, an yi nazarin tarko na gidajen da aka haɗa ta amfani da samfurin dacewa da tsarin aiki na musamman, inda jan hankali shine canjin da ya dogara da shi kuma magani (haɗin roba vs sarrafawa) shine tasirin da aka gyara (JMP® 14.0. 0. SAS Institute Inc.). A nan, mun bayar da rahoton ƙimar χ2 da p daga gwajin rabo mai yiwuwa.
Kimanta ko yana da aminci. arabiensis ta sami damar samun fitsari, babban tushen nitrogen, urea, ta hanyar ciyarwa kai tsaye, cikin awanni 48 na shan sa'o'i 4 bayan gwajin neman mai masaukin baki da kuma gwajin ciyar da mata masu shayarwa (Hoto na 1A). Duka mata masu neman mai masaukin baki da masu tsotsar jini sun sha sucrose fiye da kowace abinci ko ruwa (F(5,426) = 20.15, p < 0.0001 da F(5,299) = 56.00, p < 0.0001, bi da bi; Hoto na 1B,C). Bugu da ƙari, mata masu neman mai masaukin baki sun ci ƙasa da abinci a cikin fitsari a awanni 72 idan aka kwatanta da fitsari a awanni 168 (Hoto na 1B). Lokacin da aka ba su abinci mai ɗauke da urea, mata masu neman mai masaukin baki sun sha urea mai yawa a 2.69 mM idan aka kwatanta da duk sauran abubuwan da suka tara da ruwa, yayin da ba za a iya bambanta su da 10% sucrose ba (F(10,813) = 15.72, p < 0.0001; Hoto na 1D). Wannan ya bambanta da martanin mata masu shan jini, waɗanda galibi suna shan abinci mai ɗauke da urea fiye da ruwa, kodayake ƙasa da kashi 10% na sucrose (F(10,557) = 78.35, p < 0.0001; Hoto na 1).1E). Bugu da ƙari, lokacin da aka kwatanta tsakanin yanayin ilimin halittar jiki guda biyu, mata masu phlebotom sun sha ƙarin urea fiye da mata masu neman masauki a mafi ƙarancin yawan, kuma waɗannan mata suna shan irin wannan adadin urea a mafi yawan yawan (F(1,953)= 78.82, p < 0.0001; Hoto na 1F, G). Duk da cewa shan abinci mai ɗauke da urea ya bayyana yana da ƙimar da ta dace (Hoto na 1D,E), mata a cikin yanayin ilimin halittar jiki guda biyu sun sami damar daidaita adadin urea da ke sha a duk faɗin yawan urea ta hanyar layi ɗaya (Hoto na 1F,G). ).Hakazalika, sauro suna da ikon sarrafa shan nitrogen ta hanyar daidaita adadin fitsarin da ke sha, kamar yadda adadin nitrogen a cikin fitsari ke nunawa a cikin adadin da ke sha (Hoto na 1B, C da B insets).
Domin tantance tasirin fitsari da urea akan tsirar sauro masu neman mai masaukin baki da kuma tsotsar jini, an ba wa mata fitsari na shekaru huɗu (sabo, awanni 24, awanni 72, da awanni 168 bayan an cire su) da kuma yawan sinadarin urea, da kuma ruwan da aka tace da kuma kashi 10% na sucrose da aka yi amfani da su a matsayin iko (Hoto na 2A). Wannan nazarin rayuwa ya nuna cewa abinci yana da tasiri sosai kan rayuwar gaba daya ga mata masu neman mai masaukin baki (fitsari: χ2 = 108.5, df = 5, p < 0.0001; urea: χ2 = 122.8, df = 5, p < 0.0001; Hoto na 2B, C) da mata masu shan jini (fitsari: χ2 = 93.0, df = 5, p < 0.0001; urea: χ2 = 137.9, df = 5, p < 0.0001; Hoto na 2D,E). A cikin duka Gwaje-gwajen da aka gudanar, mata sun ci abinci mai dauke da fitsari, urea, da ruwa, inda suka rage yawan rayuwa idan aka kwatanta da mata da suka ci abinci mai dauke da sucrose (Hoto na 2B-E). Matan da ke neman masauki sun ci abinci mai dauke da fitsari mai dauke da sabo da kuma wanda ya tsufa, inda wadanda suka ci abinci mai dauke da sinadarin urea na tsawon awanni 72 (p = 0.016) suka fi karancin damar rayuwa (Hoto na 2B). Bugu da kari, matan da ke neman masauki sun ci abinci mai dauke da sinadarin urea na tsawon awanni 135 sun tsira fiye da yadda ake sarrafa ruwa (p < 0.04) (Hoto na 2C). Idan aka kwatanta da ruwa, mata da aka ci da fitsari mai dauke da sabo da kuma fitsari mai tsawon awanni 24 sun rayu tsawon lokaci (p = 0.001 da p = 0.012, bi da bi; Hoto na 2D), yayin da mata da aka ci da fitsari mai tsawon awanni 72 sun rayu tsawon lokaci fiye da wadanda aka ci da fitsari mai gajeren lokaci da kuma fitsari mai tsawon awanni 24 (p < 0.0001 da p = 0.013, bi da bi; Hoto na 2D). Lokacin da aka ci abinci mai dauke da sinadarin urea na tsawon mintuna 135, mata da aka ci da jini sun rayu tsawon lokaci fiye da duk sauran sinadarin urea da ruwa (p <0.013; Hoto na 2E).
Rayuwar mace mai masauki da tsotsar jini Anopheles arabinis suna cin fitsarin saniya da urea. A cikin nazarin halittu (A), an ba wa sauro mata abinci mai ɗauke da sabon fitsarin saniya da tsufa, yawan urea, sucrose (10%) da ruwan da aka tace (H2O). An yi rikodin rayuwar sauro masu neman mai masauki (B, C) da tsotsar jini (D, E) duk bayan sa'o'i 12 har sai dukkan mata sun ci fitsari (B, D) da urea (C, E), kuma an tabbatar da cewa suna da sinadarin Sucrose da ruwa, sun mutu.
Jimillar nisan da adadin zagayen da aka ƙayyade a gwajin injin niƙa na jirgin sama a cikin tsawon awanni 24 sun bambanta tsakanin sauro masu neman gida da kuma masu tsotsar jini, wanda hakan ya nuna ƙarancin aikin tashi gaba ɗaya (Hoto na 3). Sauro masu neman gida waɗanda ke samar da fitsari sabo da tsufa ko sucrose da ruwa sun nuna yanayin tashi daban-daban (Hoto na 3), inda mata masu cin sabon fitsari suna ƙara aiki da asuba, yayin da waɗanda aka ciyar da sauro masu shekaru 24 da 168 waɗanda suka ci fitsari sun nuna yanayin tashi daban-daban kuma galibi suna yin sa'o'i da rana. Sauro mata da ke samar da sucrose ko fitsari na awanni 72 sun nuna aiki a cikin tsawon awanni 24, yayin da mata da ke samar da ruwa sun fi aiki a tsakiyar lokacin. Sauro da aka ciyar da sucrose sun nuna mafi girman matakan aiki da daddare da safe, yayin da waɗanda suka ci fitsarin masu shekaru 72 sun sami raguwar aiki a cikin awanni 24 (Hoto na 3).
Ayyukan tashi na mace mai neman jini mai suna Anopheles arabinis da ke cin fitsarin saniya da urea. A cikin gwajin injin niƙa jirgin sama, sauro mata da aka ciyar da su da fitsarin saniya sabo da tsufa, an haɗa nau'ikan urea, sucrose (10%), da ruwan da aka tace (H2O) zuwa hannayen da ke kwance, suna juyawa cikin 'yanci (a sama). Ga mata masu neman gida (hagu) da masu tsotsar jini (dama), an rubuta jimlar nisan da adadin tashi a kowace awa ga kowane abinci a cikin tsawon awanni 24 (duhu: launin toka; haske: fari). Matsakaicin nisa da matsakaicin adadin fafatawa an nuna su a hannun dama na jadawalin ayyukan circadian. Sandunan kuskure suna wakiltar kuskuren daidaito na matsakaicin. Binciken ƙididdiga duba rubutu
Gabaɗaya, jimillar ayyukan tashi na mata masu neman masauki sun bi tsarin da ya yi kama da na nisan tashi a cikin awanni 24. Matsakaicin nisan tashi ya shafi abincin da aka ci (F(5, 138) = 28.27, p < 0.0001), kuma mata masu neman masauki sun sha fitsari na tsawon awanni 72 sun tashi nesa mai nisa sosai idan aka kwatanta da duk sauran abinci (p < 0.0001), kuma sauro masu shan sucrose sun tashi fiye da sauro sabo (p = 0.022) da sauro masu shan fitsari na tsawon awanni 24 (p = 0.022). Sabanin tsarin aikin tashi da aka bayyana ta hanyar abincin fitsari, mata masu neman masauki da ke shan urea sun nuna ci gaba da aikin tashi a cikin awanni 24, suna kololuwa a lokacin rabin lokaci na biyu na duhu (Hoto na 3). Duk da cewa tsarin aiki iri ɗaya ne, mata masu neman masauki da ke shan urea sun ƙara matsakaicin nisan tashi dangane da yawan shan ruwa (F(5, 138) = 1310.91, p < 0.0001). Mata masu neman masaukin baki suna ciyar da duk wani taro na urea fiye da yadda mata ke ciyar da ruwa ko sucrose (p < 0.03).
Jimillar aikin tashi na sauro masu tsotsar jini ya kasance mai karko kuma ya ci gaba da aiki tsawon awanni 24 a duk abincin da ake ci, tare da ƙaruwar aikin fitsari a rabin lokaci na biyu na duhu ga mata da aka ciyar da su da ruwa da kuma mata da aka ciyar da su sabo da kuma 'yan awanni 24 (hoto na 3). Duk da cewa abincin fitsari ya shafi matsakaicin nisan tashi ga mata masu shan jini (F(5, 138) = 4.83, p = 0.0004), abincin urea bai yi ba (F(5, 138) = 1.36, p = 0.24) .tare da sauran fitsari da abinci mai sarrafa kansa (sabo, p = 0.0091; awanni 72, p = 0.0022; awanni 168, p = 0.001; sucrose, p = 0.0017; dH2O, p = 0.036).
An tantance tasirin fitsari da ciyar da urea akan ma'aunin haihuwa a cikin gwaje-gwajen halittu na samar da ƙwai (Hoto na 4A) kuma an bincika su bisa ga adadin ƙwai da kowace mace ta saka, girman ƙwai, da kuma tsutsotsin farko da suka fara kyankyasa. Adadin ƙwai da aka saka. Matan Larabawa da aka ciyar da fitsari ya bambanta dangane da abinci (F(5,222) = 4.38, p = 0.0008; Hoto na 4B). Mata sun ciyar da fitsari na awanni 24, jinin da aka ciyar da sucrose ya fi ƙwai fiye da mata da aka ciyar da sauran abinci na fitsari kuma sun yi kama da waɗanda aka ciyar da sucrose (Hoto na 4B). Haka kuma, girman ƙwai da mata da aka ciyar da fitsari suka saka ya bambanta dangane da abinci (F(5, 209) = 12.85, p < 0.0001), tare da fitsari na awanni 24 da mata da aka ciyar da sucrose sun yi ƙwai mafi girma fiye da mata da aka ciyar da ruwa, yayin da ƙwai na mata da aka ciyar da sucrose na awanni 168 sun yi ƙanƙanta sosai (Hoto na 4C). Bugu da ƙari, abincin fitsari ya shafi girman tsutsotsi sosai (F(5, 187) = 7.86, p < 0.0001), tare da tsutsotsi masu girma da yawa suna fitowa daga ƙwai da mata masu sa'o'i 24 da 72 suka zubar fiye da ƙwai da aka saka daga tsutsotsi. Mata masu shayarwa da ruwa da kuma mata masu shayarwa da sa'o'i 168 (Hoto na 4D).
Ayyukan haihuwa na mata Anopheles arabinis suna ciyar da fitsarin saniya da urea. An ciyar da sauro mata da aka shayar da jini abinci wanda ya ƙunshi fitsarin saniya sabo da tsufa, yawan urea daban-daban, sucrose (10%), da ruwan da aka tace (H2O) na tsawon awanni 48 kafin a saka su a cikin gwaje-gwajen halittu da kuma samun substrates na kwanciya ƙwai awanni 48 (A). Adadin ƙwai (B, E), girman ƙwai (C, F) da girman tsutsotsi (D, G) abincin da aka bayar ya shafi sosai (fitsarin saniya: BD; urea: EG). Ma'aunin kowane ma'auni da aka auna ta amfani da sunayen haruffa daban-daban sun bambanta sosai da juna (ANOVA ta hanya ɗaya ta amfani da nazarin Tukey bayan hoc; p < 0.05). Sandunan kuskure suna wakiltar kuskuren daidaito na matsakaicin
A matsayin babban sinadarin nitrogen a cikin fitsari, urea, lokacin da aka ba ta a matsayin abinci ga mata masu shayarwa a matsayin abinci, ta shafi ma'aunin haihuwa sosai a duk nazarin. Adadin ƙwai da mata suka ciyar da urea, bayan cin abinci, ya danganta da yawan urea (F(11, 360) = 4.69; p < 0.0001), mata sun ciyar da yawan urea tsakanin 134 µM da 1.34 mM sun haifi ƙwai da yawa (Hoto na 4E). Matan da aka ciyar da yawan urea na 134 µM ko sama da haka suna haifar da ƙwai mafi girma fiye da mata da aka ciyar da ruwa (F(10, 4245) = 36.7; p < 0.0001; Hoto na 4F), da girman tsutsa, kodayake yawan urea iri ɗaya ya shafi iyaye mata (F(10, 3305) = 37.9; p < 0.0001) ya fi canzawa (Hoto na 4G).
Gabaɗaya jan hankalin da ake samu daga ruwan fitsarin shanu masu neman wurin zama. An tantance arabiensis a cikin bututun gilashi mai suna olfactometer (Hoto na 5A) sosai sakamakon shekarun fitsari (χ2 = 15.9, df = 4, p = 0.0032; Hoto na 5B). Binciken bayan hoc ya nuna cewa warin fitsari mai daɗewa a cikin awanni 24 ya haifar da ƙarin matakan jan hankali idan aka kwatanta da duk sauran jiyya (awanni 72: p = 0.0060, awanni 168: p = 0.012, pentane: p = 0.00070), Banda warin fitsari mai sabo (p = 0.13; Hoto na 5B). Kodayake jan hankalin sauro masu shan jini ga warin fitsari bai bambanta sosai ba (χ2 = 8.78, df = 4, p = 0.067; Hoto na 5C), an gano cewa waɗannan mata sun fi jan hankali ga ruwan da ake samu daga ... (p = 0.0066; Hoto na 5C).
Amsoshin ɗabi'a ga ƙamshin fitsari na shanu na halitta da na roba a cikin neman mai masaukin baki da wanda aka shayar da jini Anopheles arabianus. Tsarin bututun gilashi olfactometer (A). Jan hankalin abubuwan da ke canzawa a sararin sama daga fitsarin saniya sabo da tsufa zuwa sauro mai masaukin baki (B) da kuma masu tsotsar jini (C). Nemo martanin tentacle na Ubangiji An. An nuna abubuwan da ke fitowa daga fitsarin saniya sabo (D), awanni 24 (E), awanni 72 (F), da awanni 168 (G). Alamun gano eriya ta lantarki (EAD) suna nuna canje-canjen ƙarfin lantarki a martanin mahaɗan halittu masu aiki a cikin sararin sama da aka cire daga chromatograph ɗin gas kuma aka gano ta hanyar na'urar gano ionization na harshen wuta (FID). Sandar sikelin tana wakiltar girman amsawa (mV) idan aka kwatanta da lokacin riƙewa (s). An nuna kaddarorin da ƙimar fitarwa (µg h-1) na mahaɗan da ke aiki a zahiri. Alamar alama ɗaya (*) tana nuna amsawar ƙarancin girma. Alamun taurari biyu (**) suna nuna amsoshin da ba za a iya sake samarwa ba. Nemo Mai masaukin baki (H) da kuma mai tsotsar jini (I) An.arabiensis yana da sha'awa daban-daban ga gaurayen roba na ƙamshin fitsarin shanu sabo da na tsufa. Matsakaicin adadin sauro da ke jawo hankalin sunayen haruffa daban-daban sun bambanta sosai da juna (ANOVA ta hanya ɗaya ta amfani da nazarin Tukey bayan hoc; p < 0.05). Sandunan kuskure suna wakiltar kuskuren ma'auni na sikelin
Mace Ann.arabiensis, sa'o'i 72 da sa'o'i 120 bayan cin abinci na jini, yayin haihuwa, ba a nuna fifiko ga cirewar da ke fitowa daga fitsarin saniya sabo da tsufa ba idan aka kwatanta da magungunan pentane (χ2 = 3.07, p > 0.05; Ƙarin fayil na 1: Hoto na S1).
Ga mace Ann.arabiensis, binciken GC-EAD da GC-MS ya gano mahaɗan halittu guda takwas, shida, uku da uku masu aiki (Hoto na 5D-G). Duk da cewa an lura da bambance-bambance a cikin adadin mahaɗan da suka haifar da martanin electrophysiological, yawancin waɗannan mahaɗan suna nan a cikin kowane ruwan da aka samo daga fitsari sabo da tsufa. Saboda haka, ga kowane ruwan da aka cire, mahaɗan da suka samar da martanin ilimin halittar jiki daga antennae na mace sama da matakin da aka ƙayyade an haɗa su cikin ƙarin nazarin.
Jimlar fitowar sinadarai masu aiki a cikin tarin sararin sama ya ƙaru daga 29 µg h-1 a cikin fitsari sabo zuwa 242 µg h-1 a cikin fitsari mai tsawon sa'o'i 168, galibi saboda p-cresol da m-formaldehyde Phenol yana ƙaruwa da kuma phenol. Sabanin haka, ƙimar sakin wasu mahadi, kamar 2-cyclohexen-1-one da decanal, ya ragu tare da ƙaruwar shekarun fitsari, wanda ya yi daidai da raguwar ƙarfin sigina (yawan) da aka lura a cikin chromatogram (Hoto na 5D)-G na gefen hagu) da kuma martanin ilimin halittar jiki ga waɗannan mahadi (Hoto na 5D-G na gefen dama).
Gabaɗaya, cakudawar roba tana da irin wannan rabo na halitta na mahaɗan halitta waɗanda aka gano a cikin abubuwan da aka samo daga cikin sabbin hanyoyin fitsari da suka tsufa (Hoto na 5D–G) kuma bai bayyana ya haifar da babban jan hankali ba a cikin neman mai masaukin baki (χ2 = 8.15, df = 4, p = 0.083; Hoto na 5H) ko sauro masu tsotsar jini (χ2 = 4.91, df = 4, p = 0.30; Hoto na 5I). Duk da haka, kwatancen bayan hoc tsakanin jiyya sun nuna cewa sauro masu neman mai masaukin baki suna da matuƙar jan hankali ga cakudawar roba ta fitsari mai shekaru na awanni 24 idan aka kwatanta da sarrafa pentane (p = 0.0086; Hoto na 5H).
Domin tantance rawar da sassan jiki daban-daban ke takawa a cikin gaurayen fitsarin da aka yi da fitsari mai shekaru 24, an kimanta gaurayen da aka cire guda shida da aka cire a kan gaurayen da aka gama a gwajin Y-tube, wanda aka cire mahaɗan da aka cire a cikinsa. Ga sauro masu neman masauki, cire mahaɗan da aka cire daga cikin gaurayen da aka cire ya yi tasiri sosai kan martanin hali (χ2 = 19.63, df = 6, p = 0.0032; Ƙarin fayil 1: Hoto na S2A), duk gaurayen da aka cire sun fi kyau fiye da Ƙarami fiye da gauraye da aka haɗa gaba ɗaya. Sabanin haka, cire mahaɗan da aka cire daga cikin gaurayen da aka haɗa gaba ɗaya bai shafi halayen sauro masu tsotsar jini ba (χ2 = 11.38, df = 6, p = 0.077), ban da decanal, wanda ya haifar da ƙananan matakan idan aka kwatanta da cikakken gaurayen da aka jawo (p = 0.022; Ƙarin Fayil na 1: Hoto na S2B).
A wani ƙauye da ke fama da cutar malaria a Habasha, an kimanta ingancin haɗin fitsarin shanu na sa'o'i 24 wajen jawo sauro a ƙarƙashin yanayin fili na tsawon dare goma (Hoto na 6A). An kama kuma an gano sauro 4,861, waɗanda kashi 45.7% na su ne Anthropus.gambiae sl, kashi 18.9% na su ne Anopheles pharoensis da kashi 35.4% na su ne Culex spp. (Ƙarin fayil na 1: Tebur S1). Anopheles arabinis shine kaɗai memba na rukunin nau'ikan An.Gambian da aka gano ta hanyar binciken PCR. A matsakaici, an kama sauro 320 a kowace dare, a lokacin da tarkuna tare da haɗakar abinci suka kama sauro fiye da tarkuna biyu ba tare da haɗakar abinci ba (χ2(0, 3196) = 170.0, p < 0.0001). An sanya tarkuna marasa tarko a kowane dare biyar na gwaji a farkon, tsakiya, da ƙarshen gwajin. An kama irin wannan adadin sauro a kowane biyu tarkuna, wanda ke nuna babu son kai tsakanin gidaje (χ2(0, 1665) = 9 × 10-13, p > 0.05) kuma babu raguwar yawan jama'a a lokacin binciken. Idan aka kwatanta da tarkunan sarrafawa, adadin sauro da aka kama a cikin tarkunan da ke ɗauke da cakuda na roba ya ƙaru sosai: neman mai masaukin baki (χ2(0, 2107) = 138.7, p < 0.0001), ciyar da jini na baya-bayan nan (χ2(0, 650) = 32.2, p < 0.0001) da ciki (χ2(0, 228) = 6.27, p = 0.0123; Ƙarin fayil na 1: Tebur S1). Wannan kuma yana nuna a cikin jimlar adadin sauro da aka kama: neman mai masaukin baki > tsotsar jini > ciki > mai ciki rabin ciki > namiji.
Kimanta ingancin cakuda warin fitsari na shanu na sa'o'i 24. An gudanar da gwaje-gwajen fili a kudu maso tsakiyar Habasha (taswira), kusa da garin Maki (saka), ta amfani da tarkon haske na Cibiyar Kula da Cututtuka (CDC) (dama) a cikin gidaje biyu, tare da ƙirar murabba'in Latin (hoton iska) (A). Tarkon CDC mai wari mai wari yana jawo hankalin mata Anopheles arabesques (B), amma ba Anopheles farroes (C) ba, ta wata hanya daban, tasirin yanayin jiki. Bugu da ƙari, waɗannan tarkon sun kama adadi mai yawa na sauro na Culex mai masaukin baki. (D) Idan aka kwatanta da sarrafawa. Sandunan da ke hagu suna wakiltar matsakaicin ma'aunin zaɓi na sauro da aka kama a cikin nau'ikan tarko masu wari (kore) da tarko masu sarrafawa (buɗe) (N = 10), yayin da sandunan da ke dama suna wakiltar matsakaicin ma'aunin zaɓi a cikin tarko masu sarrafawa (buɗe; N = 5). ). Alamun alama suna nuna matakan mahimmancin ƙididdiga (*p = 0.01 da ***p < 0.0001)
An kama nau'ikan uku daban-daban a cikin tarkuna da ke ɗauke da gaurayen roba. Neman mai masaukin baki (χ2(1, 1345) = 71.7, p < 0.0001), ciyar da jini (χ2(1, 517) = 16.7, p < 0.0001) da ciki (χ2(1, 180) = 6.11, p = 0.0134) an kama .arabiensis a cikin tarkon da ke sakin gaurayen roba (Hoto na 6B), yayin da adadin An bai bambanta ba. An gano Pharoensis a cikin yanayi daban-daban na ilimin halittar jiki (Hoto na 6C). Ga Culex, an sami ƙaruwa mai yawa a yawan sauro da ke neman mai masaukin baki a cikin tarkunan da aka kama da gaurayen roba (χ2(1,1319) = 12.6, p = 0.0004; Hoto na 6D), idan aka kwatanta da tarkunan sarrafawa.
An yi amfani da tarkunan da aka yi amfani da su wajen masu masaukin baki tsakanin wuraren kiwo da kuma al'ummomin karkara a Habasha don tantance ko sauro na malaria suna amfani da warin fitsarin saniya a matsayin alamar mazaunin masu masaukin baki. Idan babu alamun masu masaukin baki, zafi, kuma tare da ko ba tare da warin fitsarin saniya ba, ba a kama sauro ba (Ƙarin fayil na 1: Hoto na S3). Duk da haka, a gaban zafin jiki mai yawa da warin fitsarin saniya, an jawo kuma an kama sauro na malaria mata, kodayake a cikin ƙananan adadi, ba tare da la'akari da shekarun fitsari ba (χ2(5, 25) = 2.29, p = 0.13; Ƙarin fayil na 1: Hoto na S3). Sabanin haka, sarrafa ruwa bai kama sauro na malaria a yanayin zafi mai yawa ba (Ƙarin Fayil na 1: Hoto na S3).
Sauro na zazzabin cizon sauro suna samun kuma suna rarraba sinadarai masu ɗauke da nitrogen ta hanyar ciyar da fitsarin saniya (watau kududdufai) don haɓaka halayen rayuwa, kamar sauran kwari [2, 4, 24, 25, 26]. Fitsari na saniya abu ne mai sauƙin samu wanda ke da alaƙa da wuraren hutawa ga masu kamuwa da cutar malaria, kamar su rumfunan shanu da ciyayi masu tsayi kusa da gidajen karkara da wuraren haihuwa. Sauro mata suna samun wannan albarkatun ta hanyar wari kuma suna iya daidaita shan sinadaran nitrogen a cikin fitsari, gami da urea, babban sinadarin nitrogen a cikin fitsari [15, 16]. Dangane da yanayin lafiyar sauro na mace, ana ware abubuwan gina jiki a cikin fitsari don haɓaka ayyukan tashi da rayuwar sauro mata masu neman masauki, da kuma halaye na rayuwa da haihuwa na mutanen da aka ciyar da jini a lokacin zagayowar gonadotropic na farko. Saboda haka, haɗa fitsari yana taka muhimmiyar rawa a cikin abinci mai gina jiki ga masu kamuwa da cutar malaria waɗanda ke rufe kamar manya masu rashin abinci mai gina jiki [8], domin yana ba sauro mata damar samun muhimman sinadarai nitrogen ta hanyar shiga cikin ƙarancin haɗari. Wannan binciken yana da manyan sakamako na annoba, yayin da mata ke ƙara tsawon rayuwarsu, aiki. da kuma fitowar haihuwa, duk waɗannan suna shafar ƙarfin vector. Bugu da ƙari, wannan ɗabi'a na iya zama abin da shirye-shiryen sarrafa vector na gaba za su mayar da hankali a kai.