Lolu cwaningo luhlole ukubulawa, ubuqili, kanye nobuthi bezentengiselwanoi-cypermethrinukwakheka kuma-anuran tadpoles. Esivivinyweni esibucayi, ukugxila kwe-100-800 μg/L kuhlolwe amahora angama-96. Esivivinyweni esingalapheki, ukugxila kwe-cypermethrin okwenzeka ngokwemvelo (1, 3, 6, kanye no-20 μg/L) kwahlolelwa ukufa, kulandelwa ukuhlolwa kwe-micronucleus kanye nokungajwayelekile kwamangqamuzana egazi abomvu enuzi izinsuku eziyi-7. I-LC50 yokwakhiwa kwe-cypermethrin yokuhweba kuma-tadpoles yayingu-273.41 μg L−1. Ekuhlolweni okungapheli, ukugxila okuphezulu kakhulu (20 μg L−1) kubangele ukufa okungaphezu kuka-50%, njengoba kubulale ingxenye yoshobishobi abahloliwe. Ukuhlolwa kwe-micronucleus kubonise imiphumela ebalulekile ku-6 kanye ne-20 μg L-1 kanye nokungajwayelekile kwe-nuclear okuningana kwatholwa, okubonisa ukuthi ukwakheka kwe-cypermethrin yokuhweba kunamandla we-genotoxic ngokumelene no-P. gracilis. I-Cypermethrin iyingozi enkulu yalolu hlobo, okubonisa ukuthi ingabangela izinkinga eziningi futhi ithinte ukuguquguquka kwalesi simiso semvelo esikhathini esifushane nesikhathi eside. Ngakho-ke, kungaphethwa ngokuthi ukwakheka kwe-cypermethrin yokuhweba kunemiphumela enobuthi ku-P. gracilis.
Ngenxa yokwanda okuqhubekayo kwemisebenzi yezolimo kanye nokusetshenziswa okujulile kweukulawula izinambuzaneizilinganiso, izilwane zasemanzini zivame ukuchayeka ezibulala-zinambuzane1,2. Ukungcoliswa kwemithombo yamanzi eduze kwamasimu ezolimo kungaba nomthelela ekuthuthukisweni nasekuphileni kwezinto eziphilayo ezingahlosiwe njengezilwane ezihlala emanzini.
Izilwane ezihlala emanzini ziya ngokuya zibaluleka ekuhlolweni kukamatikuletsheni wezemvelo. Ama-Anurans abhekwa njengama-bioindicators amahle okungcola kwemvelo ngenxa yezici zawo eziyingqayizivele njengemijikelezo yokuphila eyinkimbinkimbi, izinga lokukhula kwezibungu ngokushesha, isimo se-trophic, isikhumba esikwazi ukungena10,11, ukuncika emanzini ngokuzala12 namaqanda angavikelekile11,13,14. Ixoxo elincane lamanzi (Physalaemus gracilis), elivame ukubizwa ngokuthi isele elikhalayo, kukhonjiswe ukuthi liwuhlobo lwe-bioindicator lokungcoliswa kwezibulala-zinambuzane4,5,6,7,15. Lolu hlobo lutholakala emanzini amile, ezindaweni ezivikelwe noma ezindaweni ezinemvelo eguquguqukayo e-Argentina, e-Uruguay, eParaguay naseBrazil1617 futhi kubhekwa njengezinzile ngokwezigaba ze-IUCN ngenxa yokusatshalaliswa okubanzi nokubekezelela izindawo zokuhlala ezihlukene18.
Imiphumela ebulalayo iye yabikwa kuma-amphibians kulandela ukuchayeka ku-cypermethrin, okuhlanganisa izinguquko zokuziphatha, ze-morphological kanye ne-biochemical kuma-tadpoles23,24,25, ukufa okushintshiwe kanye nesikhathi sokuguquguquka, izinguquko ze-enzymatic, ukuncipha kwempumelelo yokuchayeka24,25, hyperactivity26, ukuvinjelwa kokusebenza kwe-cholinesterase7 kanye nezinguquko ze-282 zokubhukuda. Kodwa-ke, ucwaningo lwemiphumela ye-genotoxic ye-cypermethrin kuma-amphibians lunqunyelwe. Ngakho-ke, kubalulekile ukuhlola ukuthambekela kwezinhlobo ze-anuran ku-cypermethrin.
Ukungcoliswa kwemvelo kuthinta ukukhula nokuthuthukiswa okujwayelekile kwezilwane ezihlala emanzini nasezweni, kodwa umphumela omubi kakhulu ukulimala kwezakhi zofuzo ku-DNA okubangelwa ukuchayeka kwezibulala-zinambuzane13. Ukuhlaziywa kokuma kwamangqamuzana egazi kuyinkomba ebalulekile yezinto eziphilayo zokungcola kanye nobuthi obungaba khona bento ezilwaneni zasendle29. Ukuhlolwa kwe-micronucleus kungenye yezindlela ezisetshenziswa kakhulu zokunquma ubuthi be-genotoxicity bamakhemikhali endaweni30. Kuyindlela esheshayo, esebenzayo futhi engabizi eyinkomba enhle yokungcoliswa kwamakhemikhali ezinto eziphilayo ezinjengezilwane eziphila emanzini nasezweni31,32 futhi inganikeza ulwazi ngokuchayeka ezintweni ezingcolisa i-genotoxic33.
Inhloso yalolu cwaningo kwakuwukuhlola amandla anobuthi okwakhiwa kwe-cypermethrin yokuhweba kuma-tadpoles amancane asemanzini kusetshenziswa ukuhlolwa kwe-micronucleus kanye nokuhlola ubungozi bemvelo.
Ukufa okukhulayo (%) kwe-P. gracilis tadpoles kuvezwe ekugxilweni okuhlukile kwe-cypermethrin yokuhweba phakathi nenkathi enzima yokuhlolwa.
Ukufa okukhulayo (%) kwe-P. gracilis tadpoles kuvezwe ekugxilweni okuhlukile kwe-cypermethrin yokuhweba phakathi nokuhlolwa okungapheli.
Ukufa okuphezulu okuphawuliwe kwaba umphumela wemiphumela ye-genotoxic kuma-amphibians achayeke ekugxilweni okuhlukile kwe-cypermethrin (6 kanye ne-20 μg/L), njengoba kufakazelwa ukuba khona kwe-micronuclei (MN) nokungajwayelekile kwe-nuclear kuma-erythrocyte. Ukwakhiwa kwe-MN kubonisa amaphutha ku-mitosis futhi kuhlotshaniswa nokubophezela okungalungile kwama-chromosome kuma-microtubules, ukukhubazeka ku-protein complexes okubhekene nokuthathwa kwe-chromosome nokuthutha, amaphutha ekuhlukaniseni i-chromosome kanye namaphutha ekulungiseni umonakalo we-DNA38,39 futhi kungase kuhlotshaniswe nokucindezeleka okwenziwe nge-pesticide-oxidative40,41. Okunye okungavamile kwabonwa kukho konke ukugxila okuhloliwe. Ukwanda kokugxila kwe-cypermethrin kukhuphule ukungahambi kahle kwenuzi kuma-erythrocyte ngo-5% no-20% imithamo ephansi kakhulu (1 μg/L) kanye nephezulu (20 μg/L), ngokulandelana. Isibonelo, izinguquko ku-DNA yohlobo oluthile zingaba nemiphumela engathi sína kukho kokubili ukuphila kwesikhathi esifushane nesikhathi eside, okuholela ekwehleni kwabantu, ukushintsha ukufaneleka kokuzala, ukuzalanisa inzalo, ukulahlekelwa ukuhlukahluka kofuzo, kanye namazinga okufuduka ashintshile. Zonke lezi zici zingaba nomthelela ekusindeni kwezinhlobo zezilwane nokugcinwa kwazo42,43. Ukwakheka kokungajwayelekile kwe-erythroid kungase kubonise ukuvinjelwa kwe-cytokinesis, okuholela ekuhlukaneni kweseli okungavamile (i-erythrocyte binucleated)44,45; ama-multilobed nuclei ama-protrusions we-nuclear membrane enama-lobes amaningi46, kuyilapho okunye okungavamile kwe-erythroid kungase kuhlotshaniswe nokukhulisa i-DNA, njengezinso zenuzi/i-blebs47. Ukuba khona kwama-erythrocyte e-anucleated kungase kubonise ukuthuthwa kwe-oksijini ekhubazekile, ikakhulukazi emanzini angcolile48,49. I-Apoptosis ikhombisa ukufa kweseli50.
Ezinye izifundo nazo zibonise imiphumela ye-genotoxic ye-cypermethrin. U-Kabaña et al.51 ubonise ukuba khona kwe-micronuclei nezinguquko zenuzi njengamaseli e-binucleated namaseli e-apoptotic kumaseli e-Odontophrynus americanus ngemva kokuchayeka ekugxilweni okuphezulu kwe-cypermethrin (5000 kanye ne-10,000 μg L-1) ye-96 h. I-apoptosis eyenziwe nge-Cypermethrin iphinde yatholwa ku-P. biligonigerus52 kanye ne-Rhinella arenarum53. Le miphumela iphakamisa ukuthi i-cypermethrin inemiphumela ye-genotoxic ohlotsheni lwezinto eziphilayo zasemanzini nokuthi ukuhlolwa kwe-MN ne-ENA kungase kube inkomba yemiphumela ebulalayo kuzilwane eziphila emanzini nasezweni futhi kungase kusebenze ezinhlobonhlobo zemvelo kanye nenani labantu basendle elichayeke ezintweni ezinobuthi12.
Ukwakhiwa kwezentengiselwano kwe-cypermethrin kubangela ingozi ephezulu yemvelo (kokubili ebukhali futhi engapheli), nama-HQ adlula i-US Environmental Protection Agency (EPA) level54 engase ithinte kabi uhlobo lwezilwane uma lukhona endaweni ezungezile. Ekuhloleni ubungozi obungapheli, i-NOEC yokufa yayiyi-3 μg L-1, iqinisekisa ukuthi ukugxila okutholakala emanzini kungase kubangele ingozi kuzinhlobo55. I-NOEC ebulalayo ye-R. arenarum izibungu evezwe kwingxube ye-endosulfan ne-cypermethrin yayingu-500 μg L-1 ngemva kwamahora angu-168; leli nani lehle lafika ku-0.0005 μg L−1 ngemva kwamahora angu-336. Ababhali babonisa ukuthi uma ukuchayeka isikhathi eside, kuyancipha ukugxila okuyingozi ezinhlobonhlobo. Kubalulekile futhi ukugqamisa ukuthi amanani e-NOEC ayephezulu kunalawo e-P. gracilis ngesikhathi esifanayo sokuchayeka, okubonisa ukuthi impendulo yezinhlobo ze-cypermethrin iqondene nezinhlobo ezithile. Ngaphezu kwalokho, mayelana nokufa, inani le-CHQ le-P. gracilis ngemva kokuchayeka ku-cypermethrin lifinyelele ku-64.67, elingaphezu kwenani eliyireferensi elibekwe yi-US Environmental Protection Agency54, kanye nenani le-CHQ le-R. arenarum izibungu nalo laliphakeme kunaleli xabiso (CHQ> 388.00h) lelo liyi-338. izinhlobo eziningana zezilwane ezihlala emanzini. Uma kucatshangelwa ukuthi i-P. gracilis idinga cishe izinsuku ezingu-30 ukuqedela i-metamorphosis56, kungaphethwa ngokuthi ukugxila okuhloliwe kwe-cypermethrin kungase kube nomthelela ekwehleni kwenani labantu ngokuvimbela abantu abanaleli gciwane ukuthi bangene kumuntu omdala noma esigabeni sokuzala besebancane.
Ekuhloleni ubungozi okubaliwe kwe-micronuclei nokunye okungavamile kwe-erythrocyte yenuzi, amanani e-CHQ asukela ku-14.92 kuya ku-97.00, okubonisa ukuthi i-cypermethrin yayinengozi engaba khona ye-genotoxic ku-P. gracilis ngisho nasendaweni yayo yemvelo. Uma kucatshangelwa ukufa, ukugcwala okukhulu kwezinhlanganisela ze-xenobiotic okubekezelelwa ku-P. gracilis kwaba ngu-4.24 μg L-1. Kodwa-ke, ukugxila okuphansi njenge-1 μg/L nakho kubonise imiphumela ye-genotoxic. Leli qiniso lingase liholele ekwandeni kwenani labantu abangajwayelekile57 futhi libe nomthelela ekuthuthukisweni kanye nokuzalana kwezinhlobo zezilwane ezindaweni ezihlala kuzo, okuholela ekwehleni kwenani labantu abahlala emanzini.
Ukwakhiwa kwezentengiselwano kwesibulala-zinambuzane i-cypermethrin kubonise ubuthi obuphezulu obunamandla nobungapheli ku-P. gracilis. Amazinga aphezulu okufa abonwa, cishe ngenxa yemiphumela enobuthi, njengoba kufakazelwa ukuba khona kwe-micronuclei ne-erythrocyte nuclear abnormalities, ikakhulukazi ama-serrated nuclei, ama-lobed nuclei, nama-vesicular nuclei. Ngaphezu kwalokho, izinhlobo ezihloliwe zabonisa ukwanda kwezingozi zemvelo, kokubili ezinzima nezingamahlalakhona. Le datha, ehlanganiswe nezifundo zangaphambilini zeqembu lethu locwaningo, ibonise ukuthi ngisho nokwakhiwa kwezentengiselwano okuhlukile kwe-cypermethrin kusabangela ukwehla kwe-acetylcholinesterase (AChE) kanye nemisebenzi ye-butyrylcholinesterase (BChE) kanye ne-oxidative stress58, futhi kwaphumela ekushintsheni komsebenzi wokubhukuda kanye nokukhubazeka komlomo59 ku-P. gracilimeth formulations ye-leletha ye-letha letharinating high ubuthi kulolu hlobo. Hartmann et al. I-60 ithole ukuthi ukwakhiwa kwezentengiselwano kwe-cypermethrin kwakuyingozi kakhulu ku-P. gracilis kanye nolunye uhlobo lohlobo olufanayo (P. cuvieri) uma kuqhathaniswa nezinye izibulala-zinambuzane eziyisishiyagalolunye. Lokhu kuphakamisa ukuthi ukugxila okuvunyelwe ngokomthetho kwe-cypermethrin ukuze kuvikelwe imvelo kungase kubangele ukufa okuphezulu kanye nokwehla kwenani labantu isikhathi eside.
Kudingeka ucwaningo olwengeziwe ukuze kuhlolwe ubuthi besibulala-zinambuzane eziphila emanzini nasemanzini, njengoba ukugxila okutholakala endaweni kungase kubangele ukufa okuphezulu futhi kubangele ingozi engaba khona ku-P. gracilis. Ucwaningo ngezinhlobo eziphila emanzini nasemanzini kufanele lukhuthazwe, njengoba idatha yalezi zilwane iyindlala, ikakhulukazi ngezinhlobo zase-Brazilian.
Ukuhlolwa kobuthi obungapheli kuthathe amahora angu-168 (izinsuku ezingu-7) ngaphansi kwezimo ezimile futhi ukugxiliswa kwe-sublethal kwaba: 1, 3, 6 kanye no-20 μg ai L-1. Kuzo zombili izivivinyo, oshobishobi abangu-10 eqenjini ngalinye lokwelapha bahlolwa ngeziphindaphinda eziyisithupha, kungqikithi yama-tadpoles angama-60 ekugxiliseni ngakunye. Ngaleso sikhathi, ukwelashwa kwamanzi kuphela kwasebenza njengokulawula okungalungile. Ukusetha ngakunye kokuhlola kwakuhlanganisa indishi yengilazi eyinyumba enamandla angu-500 ml kanye nokuminyana kwe-tadpole engu-1 ngo-50 ml wesisombululo. I-flask yayimbozwe ngefilimu ye-polyethylene ukuvimbela ukuhwamuka futhi yayinomoya ngokuqhubekayo.
Amanzi ahlaziywa ngamakhemikhali ukuze kutholwe ukugcwala kwezibulala-zinambuzane ku-0, 96, kanye namahora angu-168. Ngokusho kukaSabin et al. 68 kanye noMartins et al. 69 , ukuhlaziya kwenziwa e-Pesticide Analysis Laboratory (LARP) ye-Federal University of Santa Maria kusetshenziswa i-gas chromatography ehlanganiswe ne-triple quadrupole mass spectrometry (i-Varian model 1200, Palo Alto, California, USA). Ukunqunywa komthamo wezibulala-zinambuzane emanzini kukhonjiswa njengokungeziwe (Ithebula SM1).
Ngokuhlolwa kwe-micronucleus (MNT) kanye nokuhlolwa kokungajwayelekile kwe-nuclear cell (RNA), ama-tadpoles angu-15 avela eqenjini ngalinye lokwelapha ahlaziywa. Ama-tadpoles abulawa izinzwa ngo-5% we-lidocaine (50 mg g-170) futhi amasampula egazi aqoqwa ngokubhoboza inhliziyo kusetshenziswa imijovo ye-heparinized elahlwayo. Ama-blood smear alungiswa kumaslayidi e-microscope oyinyumba, omisiwe emoyeni, ahlanganiswe ne-methanol engu-100% (4 °C) imizuzu engu-2, bese egcotshwa ngesisombululo se-Giemsa esingu-10% imizuzu engu-15 ebumnyameni. Ekupheleni kwenqubo, ama-slides ayegezwa ngamanzi acwecwe ukuze asuse ibala eliningi futhi omiswe ekamelweni lokushisa.
Okungenani ama-RBC angu-1000 avela ku-tadpole ngayinye ahlaziywa kusetshenziswa isibonakhulu esingu-100× ngenjongo engu-71 ukuze kutholwe ubukhona be-MN ne-ENA. Isamba sama-RBCs angama-75,796 asuka kuma-tadpoles ahlolwa kucatshangelwa ukugxila nokulawulwa kwe-cypermethrin. I-Genotoxicity yahlaziywa ngokwendlela kaCarrasco et al. kanye no-Fenech et al.38,72 ngokunquma imvamisa yezilonda zenuzi ezilandelayo: (1) amaseli e-nucleate: amaseli angenawo ama-nuclei; (2) amangqamuzana e-apoptotic: ukuhlukana kwezikhali zenuzi, ukufa kwamangqamuzana okuhleliwe; (3) ama-binucleate cell: amangqamuzana anama-nuclei amabili; (4) ama-nuclear buds noma ama-bleb cell: amangqamuzana anama-nuclei anezingqimba ezincane zolwelwesi lwenuzi, ama-blebs afana nama-micronuclei ngobukhulu; (5) amangqamuzana e-karyolyzed: amangqamuzana anohlaka lwenucleus kuphela ngaphandle kwezinto zangaphakathi; (6) amangqamuzana anonotshi: amangqamuzana anama-nuclei anemifantu esobala noma izinotshi ekumeni kwawo, abizwa nangokuthi ama-nucles amise okwezinso; (7) amaseli ahlanganisiwe: amangqamuzana anama-nuclear protrusion amakhulu kunama-vesicle okukhulunywe ngawo ngenhla; kanye (8) nama-microcells: amaseli anenuclei ejiyile kanye ne-cytoplasm encishisiwe. Izinguquko zaqhathaniswa nemiphumela yokulawula engemihle.
Imiphumela yokuhlolwa kobuthi obukhulu (LC50) yahlaziywa kusetshenziswa isofthiwe ye-GBasic kanye nendlela ye-TSK-Trimmed Spearman-Karber74. Idatha yokuhlola okungamahlalakhona ihlolelwe kusengaphambili ukujwayelekile kwephutha (i-Shapiro-Wilks) kanye ne-homogeneity of variance (Bartlett). Imiphumela yahlaziywa kusetshenziswa ukuhlaziya kwendlela eyodwa yokuhluka (ANOVA). Ukuhlolwa kuka-Tukey kwasetshenziswa ukuze kuqhathaniswe idatha phakathi kwabo, futhi ukuhlolwa kuka-Dunnett kwasetshenziswa ukuze kuqhathaniswe idatha phakathi kweqembu lokwelapha neqembu elingelona iqiniso.
Idatha ye-LOEC ne-NOEC yahlaziywa kusetshenziswa ukuhlolwa kuka-Dunnett. Ukuhlolwa kwezibalo kwenziwe kusetshenziswa isofthiwe ye-Statistica 8.0 (StatSoft) enezinga lokubaluleka elingu-95% (p <0.05).
Isikhathi sokuthumela: Mar-13-2025