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Ana amfani da fungicides sau da yawa a lokacin furannin 'ya'yan itace kuma suna iya yin barazanar pollinators na kwari.Koyaya, an san kadan game da yadda masu pollinators ba kudan zuma ba (misali, ƙudan zuma kaɗai, Osmia cornifrons) suna amsa lamba da tsarin fungicides waɗanda aka saba amfani da su akan apples yayin fure.Wannan gibin ilimi yana iyakance yanke shawara na tsari da ke ƙayyade amintaccen taro da lokacin feshin fungicides.Mun kimanta sakamakon cututtukan fungicides guda biyu (captan da mancozeb) da interlayer / phytosystem fungicides hudu (ciprocycline, myclobutanil, pyrostrobin da trifloxystrobin).Tasiri kan riba mai yawa, tsira, rabon jima'i da bambancin ƙwayoyin cuta.An gudanar da kimantawa ta hanyar amfani da kwayoyin halitta na baka na yau da kullum wanda aka bi da pollen a cikin nau'i uku bisa ga shawarar da aka ba da shawarar a halin yanzu don amfani da filin (1X), rabin kashi (0.5X) da ƙananan kashi (0.1X).Duk allurai na mancozeb da pyritisoline sun rage nauyin jiki sosai da tsirar tsutsa.Daga nan sai muka jera kwayoyin halittar 16S don tantance kwatankwacin ƙwayoyin cuta na mancozeb, maganin fungicide da ke da alhakin mafi girman mace-mace.Mun gano cewa bambance-bambancen ƙwayoyin cuta da yawa sun ragu sosai a cikin larvae da ake ciyar da pollen da aka yi wa mancozeb.Sakamakon dakin gwaje-gwajenmu ya nuna cewa fesa wasu magungunan kashe qwari a lokacin fure yana da illa musamman ga lafiyar O. cornifrons.Wannan bayanin yana da dacewa don yanke shawara na gudanarwa na gaba game da dorewar amfani da samfuran kariyar itacen 'ya'yan itace kuma yana aiki a matsayin tushen tsarin aiwatarwa da nufin kare masu pollinators.
An gabatar da kudan zuma mai zaman kansa Osmia cornifrons (Hymenoptera: Megachilidae) zuwa Amurka daga Japan a ƙarshen 1970s da farkon 1980s, kuma nau'in ya taka muhimmiyar rawa wajen pollinator a cikin sarrafa muhalli tun daga lokacin.Yawan jama'a na wannan kudan zuma wani yanki ne na kusan nau'ikan kudan zuma 50 waɗanda ke cike da ƙudan zuma waɗanda ke lalata gonakin almond da apple a Amurka2,3.Kudan zuma na Mason suna fuskantar ƙalubale da yawa, waɗanda suka haɗa da rarrabuwar kawuna, ƙwayoyin cuta, da magungunan kashe qwari3,4.Daga cikin magungunan kashe qwari, fungicides suna rage samun kuzari, abinci5 da kwandishan jiki6,7.Ko da yake bincike na baya-bayan nan ya nuna cewa lafiyar kudan zuma na Mason yana tasiri kai tsaye ta hanyar commensal da ectobactic microorganisms, 8,9 saboda ƙwayoyin cuta da fungi na iya yin tasiri ga abinci mai gina jiki da martanin rigakafi, tasirin bayyanar cututtukan fungicides akan nau'ikan ƙudan zuma na Mason sun fara zama. yayi karatu.
Fungicides na sakamako daban-daban (lamba da tsarin) ana fesa su a cikin gonakin gona kafin da kuma lokacin fure don magance cututtuka irin su apple scab, rot mai ɗaci, ɓacin launin ruwan kasa da powdery mildew10,11.Ana ɗaukar fungicides marasa lahani ga masu pollinators, don haka ana ba da shawarar ga masu lambu a lokacin lokacin furanni;Bayyanawa da shigar da waɗannan ƙwayoyin cuta ta ƙudan zuma sananne ne sosai, saboda yana cikin tsarin rajistar magungunan kashe qwari ta Hukumar Kare Muhalli ta Amurka da sauran hukumomin kula da ƙasa da yawa12,13,14.Koyaya, illolin fungicides akan waɗanda ba ƙudan zuma ba ba a san su ba saboda ba a buƙatar su a ƙarƙashin yarjejeniyar ba da izinin tallace-tallace a Amurka15.Bugu da kari, gaba daya babu daidaitattun ka'idoji don gwada kudan zuma guda 16,17, kuma kiyaye yankunan da ke ba da ƙudan zuma don gwaji yana da ƙalubale18.Ana ci gaba da gudanar da gwajin kudan zuma daban-daban a Turai da Amurka don nazarin illar magungunan kashe qwari a kan kudan zuma, kuma kwanan nan an samar da daidaitattun ka'idoji don O. cornifrons19.
Kudan zuma masu ƙaho monocytes ne kuma ana amfani da su ta kasuwanci a cikin amfanin gonakin carp a matsayin kari ko maye gurbin kudan zuma.Wadannan ƙudan zuma suna fitowa tsakanin Maris da Afrilu, tare da mazan da suka riga sun fito kwanaki uku zuwa hudu kafin matan.Bayan jima'i, mace tana tattara pollen da nectar don samar da jerin ƙwayoyin brood a cikin rami na tubular gida (na halitta ko na wucin gadi)1,20.Ana sanya ƙwai akan pollen a cikin sel;macen sai ta gina bangon yumbu kafin ta shirya tantanin halitta na gaba.Larvae na farko an rufe su a cikin chorion kuma suna ciyar da ruwan amfrayo.Daga na biyu zuwa na biyar instar (prepupa), tsutsa suna ciyar da pollen22.Da zarar ruwan pollen ya ƙare gaba ɗaya, tsutsa ta zama kwakwa, pupate kuma suna fitowa a matsayin manya a ɗakin ɗaki ɗaya, yawanci a ƙarshen bazara20,23.Manya suna fitowa a bazara mai zuwa.Rayuwar manya tana da alaƙa da riba mai ƙarfi (ƙara mai nauyi) dangane da cin abinci.Don haka, ingancin abinci mai gina jiki na pollen, da sauran abubuwa kamar yanayi ko kamuwa da magungunan kashe qwari, sune ke tabbatar da rayuwa da lafiya24.
Maganin kwari da fungicides da ake amfani da su kafin fure suna iya motsawa a cikin vasculature shuka zuwa digiri daban-daban, daga translaminar (misali, iya motsawa daga saman saman ganye zuwa ƙasan ƙasa, kamar wasu fungicides) 25 zuwa tasirin tsarin gaske., wanda zai iya shiga cikin kambi daga tushen, zai iya shigar da nectar na furanni apple26, inda za su iya kashe manya O. cornifrons27.Wasu magungunan kashe qwari kuma suna shiga cikin pollen, suna yin illa ga ci gaban tsutsar masara da haddasa mutuwarsu19.Sauran binciken sun nuna cewa wasu magungunan kashe qwari na iya canza yanayin gida na nau'in da ke da alaƙa da O. lignaria28.Bugu da kari, binciken dakin gwaje-gwaje da na filin da ke nuna yanayin fallasa magungunan kashe qwari (ciki har da fungicides) sun nuna cewa magungunan kashe qwari sun yi mummunan tasiri ga ilimin halittar jiki na 22 29 da rayuwar kudan zuma da wasu kudan zuma keɓe.Fursunoni iri-iri na fungicidal da aka shafa kai tsaye ga buɗe furanni a lokacin furanni na iya gurɓatar da pollen da manya suka tattara don ci gaban tsutsa, wanda har yanzu ana nazarin tasirinsa30.
An ƙara gane cewa ci gaban tsutsa yana tasiri ta hanyar pollen da ƙananan ƙwayoyin cuta na tsarin narkewa.Microbiome na zuma zuma yana rinjayar sigogi irin su mass31, canje-canje na rayuwa22 da mai sauƙi ga ƙwayoyin cuta32.Nazarin da suka gabata sun yi nazarin tasirin matakin ci gaba, abubuwan gina jiki, da muhalli akan microbiome na ƙudan zuma kaɗai.Wadannan binciken sun bayyana kamanceceniya a cikin tsari da yalwar tsutsa da kuma pollen microbiomes33, da kuma mafi yawan nau'ikan kwayoyin cutar Pseudomonas da Delftia, tsakanin nau'in kudan zuma guda daya.Duk da haka, ko da yake an danganta magungunan kashe qwari da dabarun kare lafiyar kudan zuma, ba a gano illar magungunan kashe qwari akan microbiota na tsutsa ta baki kai tsaye ba.
Wannan binciken ya gwada tasirin magungunan kashe qwari guda shida da aka yi amfani da su na zahiri da aka yi rajista don amfani da ita akan 'ya'yan itace a Amurka, gami da tuntuɓar juna da tsarin fungicides waɗanda ake gudanarwa ta baki zuwa ga masarar tsutsa asu daga gurɓataccen abinci.Mun gano cewa tuntuɓar juna da tsarin fungicides sun rage yawan nauyin kudan zuma da karuwar mace-mace, tare da mafi girman tasirin da ke hade da mancozeb da pyrithiopide.Daga nan mun kwatanta bambancin ƙananan ƙwayoyin cuta na tsutsa da ake ciyar da su akan abincin pollen da aka yi wa mancozeb da waɗanda ake ciyar da su akan tsarin kulawa.Mun tattauna yuwuwar hanyoyin da ke haifar da mace-mace da abubuwan da ke haifar da hadeddewar kwaro da shirye-shiryen sarrafa pollinator (IPPM)36.
Adult O. cornifrons overwintering a cocoons an samo su daga Cibiyar Nazarin 'ya'yan itace, Biglerville, PA, kuma an adana su a -3 zuwa 2 ° C (± 0.3 ° C).Kafin gwajin (kwakwalwa 600 a duka).A cikin watan Mayu 2022, an tura cocoons 100 O. cornifrons kowace rana zuwa cikin kofuna na filastik (cocoons 50 a kowace kofi, DI 5 cm × 15 cm tsayi) kuma an sanya goge a cikin kofuna don haɓaka buɗewa da samar da abin taunawa, rage damuwa akan dutse. kudan zuma37.Sanya kofuna na filastik guda biyu masu dauke da kwakwa a cikin kejin kwari (30 × 30 × 30 cm, BugDorm MegaView Science Co. Ltd., Taiwan) tare da masu ba da abinci 10 ml dauke da maganin sucrose 50% kuma adana na kwanaki hudu don tabbatar da rufewa da mating.23 ° C, dangi zafi 60%, photoperiod 10 l (ƙananan tsanani): 14 days.An saki mata da maza 100 da aka yi aure kowace safiya har tsawon kwanaki shida (100 a kowace rana) a cikin gida biyu na wucin gadi a lokacin furen apple mai girma (ngin tarko: nisa 33.66 × tsawo 30.48 × tsawon 46.99 cm; Ƙarin Hoto 1).An sanya shi a Arboretum na Jihar Pennsylvania, kusa da ceri (Prunus cerasus 'Eubank' Sweet Cherry Pie ™), peach (Prunus persica 'Contender'), Prunus persica 'PF 27A' Flamin Fury®), pear (Pyrus perifolia 'Olympic', Pyrus perifolia 'Shinko', Pyrus perifolia 'Shinseiki'), itacen apple coronaria (Malus coronaria) da nau'ikan itatuwan apple da yawa (Malus coronaria, Malus), itacen apple na gida 'Co-op 30' Enterprise™, Malus apple itace 'Co- Op 31′ Winecrisp™, Begonia 'Yanci', Begonia 'Golden Delicious', Begonia 'Nova Spy').Kowane gidan tsuntsu shuɗi na filastik ya dace a saman akwatunan katako guda biyu.Kowane akwatin gida yana ƙunshe da bututun takarda 800 mara komai (buɗaɗɗen karkace, 0.8 cm ID × 15 cm L) (Jonesville Paper Tube Co., Michigan) an saka shi cikin bututun cellophane mara kyau (0.7 OD duba Filogi na Filastik (T-1X matosai) suna ba da wuraren gida. .
Dukansu akwatunan gida sun fuskanci gabas kuma an rufe su da shingen lambun filastik kore (samfurin Everbilt # 889250EB12, girman buɗewa 5 × 5 cm, 0.95 m × 100 m) don hana rodent da damar tsuntsu kuma an sanya su a saman ƙasa kusa da akwatin gidan gida. kwalaye.Akwatin gida (Ƙarin Hoto 1a).Ana tattara ƙwai masu ɓarna a kowace rana ta hanyar tattara bututu 30 daga cikin gida tare da kai su dakin gwaje-gwaje.Yin amfani da almakashi, yanke a ƙarshen bututun, sannan a kwakkwance bututun karkace don fallasa sel brood.An cire ƙwai ɗaya ɗaya da pollen su ta amfani da spatula mai lanƙwasa (Kit ɗin kayan aikin Microslide, BioQuip Products Inc., California).An sanya ƙwai a kan takarda mai laushi kuma an sanya su a cikin abincin Petri na tsawon awanni 2 kafin a yi amfani da su a cikin gwaje-gwajenmu (Ƙarin Hoto 1b-d).
A cikin dakin gwaje-gwaje, mun kimanta yawan guba na baka na fungicides shida da aka yi amfani da su kafin da kuma lokacin furen apple a cikin nau'i uku (0.1X, 0.5X, da 1X, inda 1X shine alamar da aka yi amfani da shi a kowace galan 100 na ruwa / acre. Babban filin filin = maida hankali) a cikin filin)., Table 1).An maimaita kowane maida hankali sau 16 (n = 16).Biyu lamba fungicides (Table S1: mancozeb 2696.14 ppm da captan 2875.88 ppm) da hudu tsarin fungicides (Table S1: pyrithiostrobin 250.14 ppm; trifloxystrobin 110.06 ppm; myclobutanil 12dinil 75 ppm). 'ya'yan itatuwa, kayan lambu da kayan lambu na ado .Mun yi kama da pollen ta amfani da injin niƙa, an tura 0.20 g zuwa rijiya (24-riji Falcon Plate), kuma mun ƙara kuma mun haɗu da 1 μL na maganin fungicide don samar da pollen pyramidal tare da zurfin rijiyoyin 1 mm a cikin abin da aka sanya qwai.Sanya ta amfani da ƙaramin spatula (Ƙarin Hoto 1c,d).An adana faranti na falcon a zafin jiki (25°C) da 70% zafi dangi.Mun kwatanta su da tsutsa masu sarrafawa suna ciyar da abincin pollen iri ɗaya wanda aka bi da shi da ruwa mai tsabta.Mun yi rikodin mace-mace kuma mun auna nauyin tsutsa kowace rana har sai tsutsa ta kai shekarun haihuwa ta amfani da ma'auni na nazari (Fisher Scientific, daidaito = 0.0001 g).A ƙarshe, an tantance rabon jima'i ta hanyar buɗe kwakwar bayan watanni 2.5.
An fitar da DNA daga dukan O. cornifrons larvae (n = 3 a kowace yanayin jiyya, mancozeb-treated and untreated pollen) kuma mun yi nazarin nau'in nau'in microbial akan waɗannan samfurori, musamman saboda a cikin mancozeb an sami mafi yawan mace-mace a cikin larvae.karbar MnZn.An haɓaka DNA, an tsarkake ta ta amfani da DNAZymoBIOMICS®-96 MagBead DNA kit (Zymo Research, Irvine, CA), da jerin (zagaye 600) akan Illumina® MiSeq™ ta amfani da kayan v3.An yi niyya jerin abubuwan da aka yi niyya na kwayoyin 16S ribosomal RNA na kwayan cuta ta amfani da Kit ɗin Prep Library na Quick-16S™ NGS (Binciken Zymo, Irvine, CA) ta amfani da firam ɗin da ke nufin yankin V3-V4 na 16S rRNA gene.Bugu da ƙari, an yi jeri na 18S ta amfani da haɗa 10% PhiX, kuma an yi haɓakawa ta amfani da 18S001 na farko da NS4.
Shigo da aiwatar da abubuwan karantawa guda biyu ta amfani da bututun QIIME2 (v2022.11.1).An gyara waɗannan karatun kuma an haɗa su, kuma an cire jeri na chimeric ta amfani da plugin DADA2 a cikin QIIME2 (qiime dada2 hayaniyar hayaniyar)40.Ayyukan aji na 16S da 18S an yi su ta amfani da plugin classify-sklearn da aka riga aka horar da silva-138-99-nb-classifier.
An duba duk bayanan gwaji don daidaitawa (Shapiro-Wilks) da kamanni na bambance-bambance (gwajin Levene).Saboda bayanan da aka saita ba su dace da zato na ƙididdigar ƙididdiga ba kuma canji ya kasa daidaita abubuwan da suka rage, mun yi ANOVA (Kruskal-Wallis) ba tare da la'akari ba tare da dalilai guda biyu (lokacin 2, 5, da 8 kwana uku). lokaci) da fungicide] don kimanta tasirin jiyya akan sabon nauyin tsutsa, sa'an nan kuma bayan hoc nonparametric kwatanta biyu da aka yi ta amfani da gwajin Wilcoxon.Mun yi amfani da samfurin linzamin kwamfuta na gaba ɗaya (GLM) tare da rarraba Poisson don kwatanta tasirin fungicides akan rayuwa a cikin abubuwan da suka shafi fungicides guda uku41,42.Don bambance-bambancen ƙididdigar yawa, adadin bambance-bambancen jerin amplicon (ASVs) ya rushe a matakin jinsi.Kwatanta yawan bambance-bambancen tsakanin ƙungiyoyi masu amfani da 16S (matakin jinsi) da 18S dangi mai yawa an yi su ta amfani da ƙirar ƙari na gabaɗaya don matsayi, sikeli, da siffa (GAMLSS) tare da rarrabawar iyali na beta zero-inflated (BEZI), waɗanda aka tsara akan macro. .Microbiome R43 (v1.1).1).Cire nau'in mitochondrial da chloroplast kafin bincike daban-daban.Saboda matakan haraji daban-daban na 18S, kawai matakin mafi ƙanƙanta na kowane taxon ne aka yi amfani da shi don nazarin bambanta.An yi duk ƙididdigar ƙididdiga ta amfani da R (v. 3.4.3., CRAN project) (Team 2013).
Bayyanawa ga mancozeb, pyrithiostrobin, da trifloxystrobin sun rage yawan nauyin jiki a cikin O. cornifrons (Fig. 1).Wadannan tasirin an kiyaye su akai-akai don duk allurai guda uku da aka tantance (Fig. 1a-c).Cyclostrobin da myclobutanil ba su rage nauyin larvae sosai ba.
Matsakaicin sabon nau'in tsutsa mai tsutsawa wanda aka auna a maki uku a ƙarƙashin jiyya na abinci guda huɗu (ciyarwar pollen + fungicide: sarrafawa, 0.1X, 0.5X da 1X allurai).(a) Ƙananan kashi (0.1X): lokacin farko (ranar 1): χ2: 30.99, DF = 6;P <0.0001, lokaci na biyu (ranar 5): 22.83, DF = 0.0009;karo na uku;aya (rana ta 8): χ2: 28.39, DF = 6;(b) rabin kashi (0.5X): lokacin farko (ranar 1): χ2: 35.67, DF = 6;P <0.0001, lokaci na biyu (rana ɗaya).): χ2: 15.98, DF = 6;P = 0.0090;aya ta uku (ranar 8) χ2: 16.47, DF = 6;(c) Shafin ko cikakken kashi (1X): lokacin farko (ranar 1) χ2: 20.64, P = 6;P = 0.0326, lokaci na biyu (ranar 5): χ2: 22.83, DF = 6;P = 0.0009;aya ta uku (rana ta 8): χ2: 28.39, DF = 6;nonparametric bincike na bambancin.Bars suna wakiltar ma'anar ± SE na kwatancen biyu (α = 0.05) (n = 16) * P ≤ 0.05, ** P ≤ 0.001, *** P ≤ 0.0001.
A mafi ƙasƙanci (0.1X), nauyin jikin tsutsa ya ragu da 60% tare da trifloxystrobin, 49% tare da mancozeb, 48% tare da myclobutanil, da 46% tare da pyrithistrobin (Fig. 1a).Lokacin da aka fallasa zuwa rabin kashi (0.5X), nauyin jikin mancozeb larvae ya ragu da 86%, pyrithiostrobin da 52% da trifloxystrobin da 50% (Fig. 1b).Cikakken kashi (1X) na mancozeb ya rage nauyin tsutsa da 82%, pyrithiostrobin da 70%, da trifloxystrobin, myclobutanil da sangard da kusan 30% (Fig. 1c).
Yawan mace-mace ya kasance mafi girma a tsakanin tsutsa da ake ciyar da pollen da aka yi wa mancozeb, sannan pyrithiostrobin da trifloxystrobin suka biyo baya.Yawan mace-mace ya karu tare da ƙara yawan mancozeb da pyritisoline (Fig. 2; Table 2).Koyaya, yawan mace-macen masara ya ƙaru kaɗan kaɗan yayin da adadin trifloxystrobin ya karu;cyprodinil da captan ba su ƙara yawan mace-mace ba idan aka kwatanta da magungunan kulawa.
An kwatanta mace-mace na tsutsa ƙudaje bayan shan pollen da aka yi amfani da su daban-daban da magungunan kashe qwari guda shida.Mancozeb da pentopyramide sun fi dacewa da bayyanar da baki ga tsummoki na masara (GLM: χ = 29.45, DF = 20, P = 0.0059) (layi, gangara = 0.29, P <0.001; gangara = 0.24, P <0.00)).
A matsakaita, a duk jiyya, 39.05% na marasa lafiya mata ne kuma 60.95% maza ne.Daga cikin jiyya na kulawa, yawan mata ya kasance 40% a cikin duka ƙananan kashi (0.1X) da rabin kashi (0.5X), da 30% a cikin nazarin filin-kashi (1X).A cikin kashi 0.1X, a cikin larvae masu ciyar da pollen da aka bi da su tare da mancozeb da myclobutanil, 33.33% na manya mata ne, 22% na manya mata ne, 44% na manya manyan larvae mata ne, 44% na manya manyan larvae mata ne.mace, 41% na manya tsutsa mata ne, kuma masu sarrafawa sun kasance 31% (Fig. 3a).A sau 0.5 kashi, 33% na tsofaffi tsutsotsi a cikin mancozeb da pyrithiostrobin kungiyar mata ne, 36% a cikin rukunin trifloxystrobin, 41% a cikin rukunin myclobutanil, da 46% a cikin rukunin cyprostrobin.Wannan adadi ya kasance 53% a cikin rukuni.a cikin ƙungiyar captan da 38% a cikin ƙungiyar kulawa (Fig. 3b).A kashi 1X, 30% na ƙungiyar mancozeb mata ne, 36% na ƙungiyar pyrithiostrobin, 44% na ƙungiyar trifloxystrobin, 38% na ƙungiyar myclobutanil, 50% na ƙungiyar kulawa sune mata - 38.5% (Fig. 3c) .
Kashi na borers na mata da maza bayan fallasa matakin tsutsa na fungicide.(a) Karancin kashi (0.1X).(b) Rabin kashi (0.5X).(c) Yawan filin ko cikakken kashi (1X).
Binciken jerin 16S ya nuna cewa ƙungiyar ƙwayoyin cuta sun bambanta tsakanin tsutsa da aka yi amfani da su tare da mancozeb pollen da aka yi amfani da su tare da pollen da ba a kula da su ba (Fig. 4a).Ma'auni na ƙananan ƙwayoyin cuta na tsutsa da ba a kula da su ba da aka ciyar da su a kan pollen ya fi girma fiye da na tsutsa da aka ciyar akan pollen da aka yi wa mancozeb (Fig. 4b).Ko da yake bambance-bambancen da aka lura a cikin wadata tsakanin ƙungiyoyi ba su da mahimmanci a ƙididdiga, yana da mahimmanci fiye da yadda aka lura don ciyar da tsutsa a kan pollen da ba a kula da su ba (Fig. 4c).Yawan dangi ya nuna cewa microbiota na tsutsa da aka ciyar akan pollen sarrafawa ya bambanta fiye da na tsutsa da aka ciyar akan tsutsa da aka yi wa mancozeb (Fig. 5a).Binciken da aka kwatanta ya nuna kasancewar nau'in 28 a cikin sarrafawa da samfurori na mancozeb (Fig. 5b).c Bincike ta amfani da jerin 18S ya nuna babu wani bambance-bambance mai mahimmanci (Ƙarin Hoto 2).
Bayanan martaba na SAV dangane da jerin 16S an kwatanta su da wadatar Shannon kuma an lura da wadata a matakin phylum.(a) Babban bincike na daidaitawa (PCoA) dangane da tsarin al'umma gabaɗayan ƙananan ƙwayoyin cuta a cikin ciyarwar pollen ko sarrafawa (blue) da tsutsa masu ciyar da mancozeb (orange).Kowane wurin bayanai yana wakiltar samfurin dabam.An ƙididdige PCoA ta amfani da nisan Bray-Curtis na multivariate t rarrabawa.Ovals suna wakiltar matakin amincewa 80%.(b) Boxplot, albarkatun Shannon albarkatun kasa (maki) da c.Dukiya mai gani.Akwatunan akwatin suna nuna kwalaye don layin tsaka-tsaki, kewayon tsaka-tsaki (IQR), da 1.5 × IQR (n = 3).
Haɗin gwiwar al'ummomin ƙananan ƙwayoyin cuta na tsutsa da ake ciyar da su akan pollen da aka yi wa mancozeb da ba a kula da su ba.(a) Dangantaka da yawa na kwayoyin halitta suna karantawa a cikin tsutsa.(b) Taswirar zafi na al'ummomin ƙananan ƙwayoyin cuta.Delftia (rashin daidaituwa (OR) = 0.67, P = 0.0030) da Pseudomonas (OR = 0.3, P = 0.0074), Microbacterium (OR = 0.75, P = 0.0617) (OR = 1.5, P = 0.0060);Layukan taswirar zafi suna taruwa ta amfani da nisa na daidaitawa da matsakaicin haɗin kai.
Sakamakonmu ya nuna cewa bayyanar baki ga lamba (mancozeb) da tsarin (pyrostrobin da trifloxystrobin) fungicides, da ake amfani da su sosai a lokacin furanni, suna rage yawan kiba da karuwar mace-mace na masara.Bugu da kari, mancozeb ya rage girman bambance-bambance da wadatar microbiome yayin matakin farko.Myclobutanil, wani nau'in fungicides na tsarin, yana rage yawan ƙwayar tsutsa ta jiki a duk allurai uku.Wannan tasirin ya bayyana a karo na biyu (rana 5) da na uku (rana 8) maki lokaci.Ya bambanta, cyprodinil da captan ba su rage yawan kiba ko rayuwa ba idan aka kwatanta da ƙungiyar kulawa.A saninmu, wannan aikin shine na farko don tantance illar yawan amfanin gonaki na fungicides daban-daban da ake amfani da su don kare amfanin gonakin masara ta hanyar fallasa pollen kai tsaye.
Duk magungunan fungicides sun rage girman nauyin jiki sosai idan aka kwatanta da magungunan sarrafawa.Mancozeb yana da mafi girman tasiri akan ribar nauyin jikin tsutsa tare da raguwar matsakaicin 51%, sannan pyrithiostrobin ya biyo baya.Duk da haka, wasu binciken ba su bayar da rahoton illar tasirin maganin fungicides a kan matakan tsutsa44 ba.Duk da cewa dithiocarbamate biocides an nuna suna da ƙananan toxicity45, ethylene bisdithiocarbamates (EBDCS) kamar mancozeb na iya ƙasƙantar da su zuwa urea ethylene sulfide.Ganin tasirinsa na mutagenic a cikin wasu dabbobi, wannan samfurin lalata yana iya zama alhakin abubuwan da aka lura46,47.Nazarin da suka gabata sun nuna cewa samuwar ethylene thiourea yana tasiri da abubuwa kamar haɓakar zafin jiki48, matakan zafi49 da tsayin ajiyar samfur50.Yanayin ajiyar da ya dace don biocides na iya rage waɗannan illolin.Bugu da ƙari, Hukumar Kula da Abinci ta Turai ta nuna damuwa game da gubar pyrithiopide, wanda aka nuna yana da ciwon daji ga tsarin narkewa na sauran dabbobi51.
Gudanar da baki na mancozeb, pyrithiostrobin, da trifloxystrobin yana ƙara yawan mace-mace na tsutsa mai ƙwayar masara.Sabanin haka, myclobutanil, ciprocycline da captan ba su da tasiri akan mace-mace.Wadannan sakamakon sun bambanta da na Ladurner et al.52, wanda ya nuna cewa captan ya rage yawan rayuwar balagagge O. lignaria da Apis mellifera L. (Hymenoptera, Apisidae).Bugu da kari, an gano magungunan kashe qwari irin su captan da boscalid suna haifar da mace-macen tsutsa52,53,54 ko canza halin ciyarwa55.Wadannan sauye-sauye, bi da bi, na iya shafar ingancin abinci mai gina jiki na pollen da kuma samun karfin kuzari na matakin tsutsa.Mutuwar da aka gani a cikin rukunin kulawa ya dace da sauran karatun 56,57.
Za'a iya bayyana ma'aunin jima'i na jima'i na maza da aka lura a cikin aikinmu ta hanyar dalilai irin su rashin isasshen mating da rashin yanayin yanayi a lokacin furanni, kamar yadda aka ba da shawara ga O. cornuta ta Vicens da Bosch.Kodayake mata da maza a cikin bincikenmu suna da kwanaki huɗu don saduwa da juna (lokacin da ake ganin ya isa don cin nasara), mun rage ƙarfin haske da gangan don rage damuwa.Koyaya, wannan gyare-gyare na iya tsoma baki tare da tsarin jima'i ba da gangan ba61.Bugu da kari, ƙudan zuma suna fuskantar kwanaki da yawa na yanayi mara kyau, gami da ruwan sama da ƙananan zafin jiki (<5°C), wanda kuma zai iya yin mummunar tasiri ga nasarar mating4,23.
Kodayake bincikenmu ya mayar da hankali kan microbiome gabaɗaya, sakamakonmu yana ba da haske game da yuwuwar alaƙa tsakanin al'ummomin ƙwayoyin cuta waɗanda ke da mahimmanci ga abincin kudan zuma da bayyanar cututtukan fungi.Misali, larvae da ke ciyar da pollen da aka yi wa mancozeb ya ragu sosai a tsarin al'umma da yawa idan aka kwatanta da tsutsa da ke ciyar da pollen da ba a kula da su ba.A cikin tsutsa da ke cinye pollen da ba a kula da su ba, ƙungiyoyin ƙwayoyin cuta Proteobacteria da Actinobacteria sun kasance masu rinjaye kuma sun fi yawan iska ko kuma aerobic.Kwayoyin Delft, yawanci suna haɗuwa da nau'in kudan zuma guda ɗaya, an san su da aikin ƙwayoyin cuta, wanda ke nuna yiwuwar kariya daga cututtuka.Wani nau'in kwayoyin cuta, Pseudomonas, yana da yawa a cikin tsutsa da ke ciyar da pollen da ba a kula da su ba, amma an rage shi sosai a cikin tsutsa masu maganin mancozeb.Sakamakonmu yana goyan bayan binciken da ya gabata wanda ke gano Pseudomonas a matsayin ɗayan mafi yawan nau'ikan halitta a cikin O. bicornis35 da sauran wasps34 kaɗai.Ko da yake ba a yi nazarin shaidar gwaji don rawar Pseudomonas a cikin lafiyar O. cornifrons ba, an nuna wannan kwayar cutar don inganta haɓakar ƙwayoyin kariya a cikin ƙwanƙwasa Paederus fuscipes da kuma inganta metabolism na arginine a cikin vitro 35, 65. Wadannan abubuwan lura sun nuna rawar da za ta taka a cikin kariya ta kwayar cuta da kwayoyin cuta a lokacin ci gaba na O. cornifrons larvae.Microbacterium wani nau'in halitta ne da aka gano a cikin bincikenmu wanda aka ruwaito yana kasancewa a cikin adadi mai yawa a cikin larvae na baƙar fata masu tashi a ƙarƙashin yanayin yunwa66.A cikin O. cornifrons larvae, microbacteria na iya taimakawa wajen daidaitawa da juriya na microbiome na gut a ƙarƙashin yanayin damuwa.Bugu da ƙari, ana samun Rhodococcus a cikin O. cornifrons larvae kuma an san shi don iyawar detoxification67.Hakanan ana samun wannan nau'in a cikin gut na A. florea, amma cikin ƙarancin yawa68.Sakamakonmu yana nuna kasancewar bambance-bambancen kwayoyin halitta da yawa a cikin yawan harajin ƙananan ƙwayoyin cuta waɗanda zasu iya canza tsarin rayuwa a cikin tsutsa.Koyaya, ana buƙatar ingantaccen fahimtar bambancin aikin O. cornifrons.
A taƙaice, sakamakon ya nuna cewa mancozeb, pyrithiostrobin, da trifloxystrobin sun rage nauyin jiki da kuma ƙara yawan mace-mace na tsutsa mai tsutsar masara.Ko da yake ana ƙara damuwa game da illolin fungicides akan masu pollinators, akwai buƙatar ƙara fahimtar tasirin ragowar metabolites na waɗannan mahadi.Ana iya shigar da waɗannan sakamakon cikin shawarwari don haɗaɗɗun shirye-shiryen sarrafa pollinator waɗanda ke taimaka wa manoma su guji yin amfani da wasu fungicides kafin da kuma lokacin furen bishiyar ta hanyar zaɓin fungicides da bambanta lokacin aikace-aikacen, ko kuma ta ƙarfafa yin amfani da wasu hanyoyin da ba su da lahani 36. Wannan bayanin yana da mahimmanci don haɓaka shawarwari.akan amfani da magungunan kashe qwari, kamar daidaita shirye-shiryen feshin da ake dasu da canza lokacin feshi lokacin zabar kayan aikin fungicides ko haɓaka amfani da wasu hanyoyin da ba su da haɗari.Ana buƙatar ƙarin bincike game da illar cututtukan fungicides akan rabon jima'i, halayyar ciyarwa, gut microbiome, da hanyoyin ƙwayoyin ƙwayoyin cuta waɗanda ke haifar da asarar nauyi da mace-mace.
Bayanan tushe 1, 2 da 3 a cikin Figures 1 da 2 an adana su a cikin ma'ajin bayanan figshare DOI: https://doi.org/10.6084/m9.figshare.24996245 da https://doi.org/10.6084/m9.figshare.24996233.Lissafin da aka bincika a cikin binciken na yanzu (Figs. 4, 5) suna samuwa a cikin ma'ajin NCBI SRA a ƙarƙashin lambar shiga PRJNA1023565.
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