Wannan binciken ya kimanta kisa, rashin ƙarfi, da guba na kasuwancicypermethrinformulations to anuran tadpoles. A cikin gwaji mai tsanani, an gwada ƙididdiga na 100-800 μg / L don 96 h. A cikin gwaji na yau da kullun, abubuwan da ke faruwa a zahiri na cypermethrin (1, 3, 6, da 20 μg/L) an gwada su don mace-mace, sannan gwajin micronucleus da cututtukan ƙwayoyin cuta na ƙwayoyin jini na jini na kwanaki 7. LC50 na ƙirar cypermethrin na kasuwanci zuwa tadpoles shine 273.41 μg L-1. A cikin gwaji na yau da kullum, mafi girman ƙaddamarwa (20 μg L-1) ya haifar da fiye da 50% mace-mace, yayin da ya kashe rabin tadpoles da aka gwada. Gwajin micronucleus ya nuna sakamako mai mahimmanci a 6 da 20 μg L-1 kuma an gano nau'o'in makaman nukiliya da yawa, yana nuna cewa samfurin cypermethrin na kasuwanci yana da yiwuwar genotoxic akan P. gracilis. Cypermethrin babban haɗari ne ga wannan nau'in, yana nuna cewa zai iya haifar da matsaloli masu yawa kuma yana shafar yanayin wannan yanayin a cikin gajeren lokaci da kuma dogon lokaci. Sabili da haka, ana iya ƙaddamar da cewa samfuran cypermethrin na kasuwanci suna da tasiri mai guba akan P. gracilis.
Saboda ci gaba da fadada ayyukan noma da aikace-aikace mai zurfi nasarrafa kwaroMa'auni, dabbobin ruwa suna yawan fuskantar magungunan kashe qwari1,2. Gurbacewar albarkatun ruwa a kusa da filayen noma na iya yin tasiri ga ci gaba da rayuwan halittun da ba su da manufa kamar su amphibians.
Amphibians suna ƙara zama mai mahimmanci a cikin ƙima na matrices muhalli. Anurans ana daukar su ne mai kyau bioindicators na muhalli gurbatawa saboda su musamman halaye kamar hadaddun rayuwa cycles, m tsutsa girma rates, trophic matsayi, permeable fata10,11, dogara ga ruwa don haifuwa12 da kuma unprotected qwai11,13,14. Dan kankanin kwadon ruwa (Physalaemus gracilis), wanda akafi sani da kukan kukan, an nuna shi a matsayin nau'in mai nuna nau'in gurbataccen maganin kashe kwari4,5,6,7,15. Ana samun nau'in nau'in a cikin ruwa mai tsayi, wurare masu kariya ko yankunan da ke da madaidaicin wurin zama a Argentina, Uruguay, Paraguay da Brazil1617 kuma ana la'akari da su barga ta hanyar rarraba IUCN saboda yawan rarrabawa da kuma jurewar wuraren zama daban-daban18.
An ba da rahoton sakamako na sublethal a cikin amphibians biyo bayan bayyanar cypermethrin, ciki har da halayen halayen, morphological da canje-canje na biochemical a cikin tadpoles23,24,25, canza mace-mace da lokacin metamorphosis, canje-canjen enzymatic, rage nasarar hatching24,25, hyperactivity26, hanawa na cholinesterase aiki27 da canje-canje a cikin aikin swimming27. Duk da haka, nazarin sakamakon genotoxic na cypermethrin a cikin amphibians yana da iyaka. Sabili da haka, yana da mahimmanci don tantance yiwuwar nau'in anuran zuwa cypermethrin.
Gurbacewar muhalli yana shafar ci gaban al'ada da ci gaban amphibian, amma mafi munin illa shine lalacewar kwayoyin halittar DNA da ke haifar da fallasa magungunan kashe qwari13. Binciken Halittar Halittar Jini muhimmin ma'anar halitta ne na gurɓatawa da yuwuwar gubar wani abu ga nau'in daji29. Gwajin micronucleus na ɗaya daga cikin hanyoyin da aka fi amfani da su don ƙayyadaddun cututtukan ƙwayoyin cuta a cikin muhalli30. Hanya ce mai sauri, inganci kuma mara tsada wacce ke da kyaun nuni ga gurbatar sinadarai na kwayoyin halitta irin su amphibians31,32 kuma yana iya ba da bayani kan fallasa ga gurɓataccen gurɓataccen abu33.
Makasudin wannan binciken shine don kimanta yiwuwar mai guba na ƙirar cypermethrin na kasuwanci zuwa ƙananan tadpoles na ruwa ta amfani da gwajin micronucleus da kimanta haɗarin muhalli.
Tarin mace-mace (%) na P. gracilis tadpoles da aka fallasa zuwa nau'ikan cypermethrin na kasuwanci daban-daban a lokacin babban lokacin gwajin.
Tarin mace-mace (%) na P. gracilis tadpoles da aka fallasa zuwa nau'ikan cypermethrin na kasuwanci daban-daban yayin gwaji na yau da kullun.
Yawan mace-mace da aka lura da shi shine sakamakon sakamakon genotoxic a cikin amphibians da aka fallasa zuwa nau'i daban-daban na cypermethrin (6 da 20 μg / L), kamar yadda aka tabbatar da kasancewar micronuclei (MN) da kuma rashin daidaituwa na nukiliya a cikin erythrocytes. Samar da MN yana nuna kurakurai a cikin mitosis kuma yana da alaƙa da ƙarancin ɗaurin chromosomes zuwa microtubules, lahani a cikin hadaddun furotin da ke da alhakin ɗaukar chromosome da jigilar kaya, kurakurai a cikin rarrabuwar chromosome da kurakurai a cikin lalacewar DNA na gyare-gyare38,39 kuma yana iya kasancewa yana da alaƙa da ƙwayar cuta ta haifar da oxidative stress40,41. An lura da wasu rashin daidaituwa a duk abubuwan da aka kimanta. Ƙara yawan adadin cypermethrin ya ƙãra rashin daidaituwa na nukiliya a cikin erythrocytes da 5% da 20% a mafi ƙasƙanci (1 μg / L) da mafi girma (20 μg / L), bi da bi. Misali, canje-canje a cikin DNA na nau'in jinsin na iya haifar da mummunan sakamako ga rayuwa na ɗan gajeren lokaci da na dogon lokaci, wanda ke haifar da raguwar yawan jama'a, canza yanayin haifuwa, ƙirƙira, hasarar bambance-bambancen kwayoyin halitta, da canjin ƙaura. Duk waɗannan abubuwan na iya yin tasiri ga rayuwar jinsuna da kiyayewa42,43. Samuwar rashin daidaituwa na erythroid na iya nuna wani toshe a cikin cytokinesis, wanda ya haifar da rarrabawar kwayoyin halitta (binucleated erythrocytes) 44,45; Multilobed nuclei ne protrusions na nukiliya membrane tare da mahara lobes46, yayin da sauran erythroid rashin lafiyan iya zama hade da DNA karawa, kamar nukiliya kodan/blebs47. Kasancewar anucleated erythrocytes na iya nuna rashin isashshen iskar oxygen, musamman a cikin gurbataccen ruwa48,49. Apoptosis yana nuna mutuwar tantanin halitta50.
Sauran nazarin kuma sun nuna tasirin genotoxic na cypermethrin. Kabaña et al.51 ya nuna kasancewar micronuclei da canje-canje na nukiliya irin su binucleated sel da ƙwayoyin apoptotic a cikin Odontophrynus americanus Kwayoyin bayan da aka yi amfani da shi zuwa babban adadin cypermethrin (5000 da 10,000 μg L-1) don 96 h. An kuma gano apoptosis mai haifar da Cypermethrin a cikin P. biligonigerus52 da Rhinella arenarum53. Wadannan sakamakon sun nuna cewa cypermethrin yana da tasirin genotoxic a kan nau'in kwayoyin halittu masu ruwa da kuma cewa MN da ENA assay na iya zama mai nuna alamar tasiri a kan amphibians kuma yana iya dacewa da nau'in asali da kuma yawan daji da aka fallasa ga masu guba12.
Shirye-shiryen kasuwanci na cypermethrin yana haifar da haɗari mai girma na muhalli (dukansu mai tsanani da na yau da kullum), tare da HQs sun wuce matakin Hukumar Kare Muhalli ta Amurka (EPA) matakin54 wanda zai iya yin mummunar tasiri ga nau'in idan akwai a cikin muhalli. A cikin ƙididdigar haɗari na yau da kullum, NOEC don mace-mace shine 3 μg L-1, yana tabbatar da cewa yawan abubuwan da aka samu a cikin ruwa na iya haifar da haɗari ga nau'in55. NOEC mai kisa na R. arenarum larvae da aka fallasa zuwa cakuda endosulfan da cypermethrin shine 500 μg L-1 bayan 168 h; wannan ƙimar ta ragu zuwa 0.0005 μg L-1 bayan 336 h. Marubutan sun nuna cewa tsawon lokacin bayyanarwa, raguwar abubuwan da ke da lahani ga nau'in. Har ila yau, yana da mahimmanci a nuna cewa ƙimar NOEC ta kasance mafi girma fiye da na P. gracilis a lokaci guda na nunawa, yana nuna cewa amsawar jinsuna ga cypermethrin shine nau'i-nau'i. Bugu da ƙari kuma, dangane da mace-mace, ƙimar CHQ na P. gracilis bayan da aka fallasa zuwa cypermethrin ya kai 64.67, wanda ya fi girma fiye da ƙimar da aka tsara ta Hukumar Kare Muhalli ta Amurka54, kuma darajar CHQ na R. arenarum larvae kuma ya fi girma fiye da wannan darajar (CHQ> 388.00) a cikin 36 da aka yi nazari a cikin haɗari mai girma. nau'ikan amphibian da yawa. Yin la'akari da cewa P. gracilis yana buƙatar kimanin kwanaki 30 don kammala metamorphosis56, ana iya ƙaddamar da cewa ƙididdigar binciken na cypermethrin na iya taimakawa wajen rage yawan jama'a ta hanyar hana masu kamuwa da cutar shiga cikin manya ko matakin haihuwa tun suna ƙanana.
A cikin ƙididdige ƙididdigar haɗarin micronuclei da sauran erythrocyte makaman nukiliya, ƙimar CHQ sun kasance daga 14.92 zuwa 97.00, yana nuna cewa cypermethrin yana da haɗarin genotoxic ga P. gracilis har ma a cikin mazauninsa. Yin la'akari da mace-mace, matsakaicin matsakaicin matakan xenobiotic mahadi masu haƙuri ga P. gracilis shine 4.24 μg L-1. Duk da haka, ƙididdiga masu ƙarancin 1 μg / L kuma sun nuna tasirin genotoxic. Wannan hujja na iya haifar da karuwa a yawan mutane marasa kyau57 kuma suna shafar haɓakawa da haifuwa na nau'in a cikin mazauninsu, wanda zai haifar da raguwa a cikin al'ummar amphibian.
Shirye-shiryen kasuwanci na cypermethrin na maganin kwari ya nuna rashin lafiya mai tsanani da na kullum zuwa P. gracilis. An lura da adadin mace-mace mafi girma, mai yiwuwa saboda sakamako masu guba, kamar yadda aka nuna ta kasancewar micronuclei da erythrocyte rashin daidaituwa na nukiliya, musamman serrated nuclei, lobed nuclei, da vesicular nuclei. Bugu da ƙari, nau'in da aka yi nazari ya nuna ƙarar haɗarin muhalli, duka mai tsanani da na yau da kullum. Wadannan bayanan, hade da binciken da aka yi a baya ta ƙungiyar binciken mu, sun nuna cewa har ma daban-daban na kasuwanci na cypermethrin har yanzu ya haifar da raguwar acetylcholinesterase (AChE) da kuma ayyukan butyrylcholinesterase (BChE) da damuwa na oxyidative58, kuma ya haifar da canje-canje a cikin aikin yin iyo da rashin daidaituwa na baki59 a cikin P. gracilis, wanda ke nuna cewa tallace-tallace da ƙaddamarwa na karuwanci suna da babban tsari na lethalcy. nau'in. Hartmann et al. 60 ya gano cewa samfuran kasuwanci na cypermethrin sun kasance mafi guba ga P. gracilis da wani nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in nau'in cypermethrin). Wannan yana nuna cewa adadin cypermethrin da aka amince bisa doka don kariyar muhalli na iya haifar da mace-mace da raguwar yawan jama'a na dogon lokaci.
Ana buƙatar ƙarin nazarin don tantance yawan guba na magungunan kashe qwari ga masu amphibians, kamar yadda yawancin da aka samu a cikin yanayi na iya haifar da mace-mace mai yawa kuma yana haifar da haɗari ga P. gracilis. Ya kamata a karfafa bincike kan nau'in amphibian, saboda bayanai kan wadannan kwayoyin halitta ba su da yawa, musamman kan nau'in Brazil.
Gwajin guba na yau da kullun na tsawon sa'o'i 168 (kwanaki 7) a ƙarƙashin yanayin da ba a iya gani ba kuma abubuwan da suka fi dacewa sune: 1, 3, 6 da 20 μg ai L-1. A cikin gwaje-gwajen guda biyu, 10 tadpoles a kowace ƙungiyar kulawa an kimanta su tare da maimaita guda shida, don jimlar 60 tadpoles a kowane taro. A halin yanzu, maganin kawai na ruwa yana aiki azaman iko mara kyau. Kowane saitin gwaji ya ƙunshi kwanon gilashi mara kyau tare da ƙarfin 500 ml da yawa na tadpole 1 a kowace 50 ml na bayani. An rufe flask ɗin da fim ɗin polyethylene don hana fitar da iska kuma ana ci gaba da yin iska.
An yi nazarin ruwa ta hanyar sinadarai don tantance yawan magungunan kashe qwari a 0, 96, da 168 h. A cewar Sabin et al. 68 da Martins et al. 69, an yi nazarin binciken a Laboratory Analysis Pestide (LARP) na Jami'ar Tarayya ta Santa Maria ta yin amfani da chromatography na gas tare da nau'in nau'i na nau'i na hudu (Varian model 1200, Palo Alto, California, USA). Ƙididdigar ƙayyadaddun ƙayyadaddun magungunan kashe qwari a cikin ruwa ana nuna su azaman ƙarin kayan (Table SM1).
Don gwajin micronucleus (MNT) da gwajin rashin daidaituwa na ƙwayoyin cuta (RNA), an bincika tadpoles 15 daga kowace rukunin jiyya. An yi amfani da tadpoles tare da 5% lidocaine (50 mg g-170) kuma an tattara samfuran jini ta hanyar huɗar zuciya ta amfani da sirinji mai zubar da ciki. An shirya smears na jini akan faifan microscope mara kyau, busasshen iska, an gyara shi da methanol 100% (4 ° C) na mintuna 2, sannan a shafa tare da maganin 10% Giemsa na mintuna 15 a cikin duhu. A ƙarshen tsari, an wanke nunin faifai tare da ruwa mai tsafta don cire tabo mai yawa kuma a bushe a zafin jiki.
Aƙalla 1000 RBCs daga kowane tadpole an bincika ta amfani da microscope 100 × tare da maƙasudin 71 don sanin kasancewar MN da ENA. An kimanta jimlar 75,796 RBCs daga tadpoles la'akari da ƙima da sarrafawa na cypermethrin. An yi nazarin Genotoxicity bisa ga hanyar Carrasco et al. da Fenech et al.38,72 ta hanyar ƙayyade yawan nau'in cututtukan nukiliya masu zuwa: (1) kwayoyin halitta: sel ba tare da nuclei; (2) Kwayoyin apoptotic: rarrabuwar nukiliya, mutuwar kwayar halitta; (3) Kwayoyin binucleate: Kwayoyin da ke da tsakiya guda biyu; (4) ƙwayoyin cuta na nukiliya ko ƙwayoyin bleb: ƙwayoyin da ke da ƙananan ƙwayoyin cuta tare da ƙananan haɓaka na membrane na nukiliya, blebs masu kama da girman micronuclei; (5) Kwayoyin karyolyzed: Kwayoyin da ke da jigon tsakiya kawai ba tare da kayan ciki ba; (6) Kwayoyin da aka ɗora: Kwayoyin da ke da ƙwaya masu tsattsauran ra'ayi ko ƙima a cikin surarsu, wanda kuma ake kira ƙwayoyin ƙwayoyin koda; (7) Kwayoyin lobulated: Kwayoyin da ke da haɓakar nukiliya sun fi girma fiye da vesicles da aka ambata; da (8) microcells: Kwayoyin da ke da dunƙule ƙwayoyin cuta da rage cytoplasm. An kwatanta canje-canjen tare da sakamakon sarrafawa mara kyau.
An tantance sakamakon gwajin guba mai tsanani (LC50) ta amfani da software na GBasic da TSK-Trimmed Spearman-Karber method74. An riga an gwada bayanan gwajin na yau da kullun don kuskuren al'ada (Shapiro-Wilks) da daidaituwar bambancin (Bartlett). An yi nazarin sakamakon ta amfani da bincike na bambance-bambancen hanya ɗaya (ANOVA). An yi amfani da gwajin Tukey don kwatanta bayanai a tsakanin juna, kuma an yi amfani da gwajin Dunnett don kwatanta bayanai tsakanin rukunin jiyya da kuma ƙungiyar kulawa mara kyau.
An yi nazarin bayanan LOEC da NOEC ta amfani da gwajin Dunnett. An yi gwajin ƙididdiga ta amfani da software na Statistica 8.0 (StatSoft) tare da matakin mahimmanci na 95% (p <0.05).
Lokacin aikawa: Maris 13-2025