Wannan bincike ya tantance mutuwa, ƙarancin mutuwa, da kuma gubar kasuwancicypermethrinTsarin da aka yi wa ƙwayoyin anuran. A cikin gwajin gaggawa, an gwada yawan ƙwayoyin 100–800 μg/L na tsawon awanni 96. A cikin gwajin na yau da kullun, an gwada yawan ƙwayoyin cypermethrin (1, 3, 6, da 20 μg/L) na halitta don mace-mace, sannan aka biyo bayan gwajin micronucleus da kuma rashin daidaituwar ƙwayoyin ja na ƙwayoyin jini na tsawon kwanaki 7. LC50 na tsarin cypermethrin na kasuwanci ga ƙwayoyin tadpoles shine 273.41 μg L−1. A cikin gwajin na yau da kullun, mafi girman yawan ƙwayoyin (20 μg L−1) ya haifar da mace-mace sama da 50%, yayin da ya kashe rabin ƙwayoyin tadpoles da aka gwada. Gwajin micronucleus ya nuna sakamako mai mahimmanci a 6 da 20 μg L−1 kuma an gano wasu bambance-bambancen nukiliya, wanda ke nuna cewa tsarin cypermethrin na kasuwanci yana da yuwuwar guba ga P. gracilis. Cypermethrin babban haɗari ne ga wannan nau'in, wanda ke nuna cewa yana iya haifar da matsaloli da yawa kuma yana shafar yanayin wannan yanayin a cikin ɗan gajeren lokaci da na dogon lokaci. Saboda haka, za a iya kammala da cewa tsarin cypermethrin na kasuwanci yana da illa mai guba ga P. gracilis.
Saboda ci gaba da faɗaɗa ayyukan noma da kuma amfani da su sosaimaganin kwarimatakan da aka ɗauka, dabbobin ruwa suna fuskantar barazanar kashe kwari1,2. Gurɓatar albarkatun ruwa kusa da filayen noma na iya shafar ci gaba da rayuwar halittu marasa manufa kamar su tsuntsayen ruwa.
Masu ruwa da tsaki suna ƙara zama masu mahimmanci wajen tantance matrices na muhalli. Ana ɗaukar Anurans a matsayin masu kyau na abubuwan da ke haifar da gurɓataccen muhalli saboda halaye na musamman kamar zagayowar rayuwa mai rikitarwa, saurin girma na tsutsotsi, yanayin trophic, fata mai shiga ciki10,11, dogaro da ruwa don haifuwa12 da ƙwai marasa kariya11,13,14. Ƙaramin kwaɗin ruwa (Physaalamus gracilis), wanda aka fi sani da kwaɗin kuka, an nuna cewa nau'in halittu ne na gurɓataccen magungunan kashe kwari4,5,6,7,15. Ana samun nau'in a cikin ruwa mai tsayi, wurare masu kariya ko yankuna masu bambancin muhalli a Argentina, Uruguay, Paraguay da Brazil1617 kuma ana ɗaukarsa a matsayin tsayayye ta hanyar rarrabuwar IUCN saboda yawan yaɗuwarta da kuma jure wa wurare daban-daban18.
An ruwaito tasirin da ke cikin ƙwayoyin halittar jiki a cikin ƙwayoyin halittar jiki bayan fallasa su ga cypermethrin, gami da canje-canjen ɗabi'a, yanayin jiki da na sinadarai a cikin tadpoles23,24,25, canjin mace-mace da lokacin metamorphosis, canje-canjen enzymatic, raguwar nasarar kyankyaso24,25, yawan aiki26, hana ayyukan cholinesterase27 da canje-canje a cikin aikin ninkaya7,28. Duk da haka, nazarin tasirin kwayoyin halitta na cypermethrin a cikin ƙwayoyin halittar jiki yana da iyaka. Saboda haka, yana da mahimmanci a tantance yuwuwar nau'in anuran ga cypermethrin.
Gurɓatar muhalli tana shafar girma da ci gaban halittu masu rai, amma mummunan tasirin shine lalacewar kwayoyin halitta ga DNA wanda ke faruwa sakamakon fallasa ƙwayoyin cuta13. Binciken yanayin ƙwayoyin jini muhimmin alamar halittu ne na gurɓatawa da yuwuwar guba ga wani abu ga nau'in daji29. Gwajin micronucleus yana ɗaya daga cikin hanyoyin da aka fi amfani da su don tantance gubar sinadarai a cikin muhalli30. Hanya ce mai sauri, inganci kuma mai araha wacce ke nuna kyakkyawan gurɓatar sinadarai na halittu kamar amphibians31,32 kuma tana iya samar da bayanai kan fallasa ga gurɓatattun halittu masu rai33.
Manufar wannan binciken ita ce a tantance yuwuwar gubar da ke tattare da tsarin cypermethrin na kasuwanci ga ƙananan tadpoles na ruwa ta amfani da gwajin micronucleus da kuma kimanta haɗarin muhalli.
Jimlar mace-mace (%) na P. gracilis tadpoles da aka fallasa su ga yawan cypermethrin na kasuwanci daban-daban a lokacin gwajin.
Jimlar mace-mace (%) na P. gracilis tadpoles da aka fallasa su ga yawan cypermethrin na kasuwanci daban-daban yayin gwaji na yau da kullun.
Yawan mace-mace da aka lura ya faru ne sakamakon tasirin kwayoyin halitta a cikin amphibians da aka fallasa su ga yawan cypermethrin daban-daban (6 da 20 μg/L), kamar yadda aka tabbatar ta hanyar kasancewar micronuclei (MN) da rashin daidaituwar nukiliya a cikin erythrocytes. Samuwar MN yana nuna kurakurai a cikin mitosis kuma yana da alaƙa da rashin daurewar chromosomes zuwa microtubules, lahani a cikin hadaddun furotin da ke da alhakin ɗaukar chromosome da jigilar su, kurakurai a cikin rabuwar chromosome da kurakurai a cikin gyaran lalacewar DNA38,39 kuma yana iya alaƙa da damuwa ta oxidative da magungunan kashe ƙwari ke haifarwa40,41. An lura da wasu rashin daidaituwa a duk yawan da aka kimanta. Ƙara yawan cypermethrin ya ƙara rashin daidaituwar nukiliya a cikin erythrocytes da 5% da 20% a mafi ƙarancin allurai (1 μg/L) da mafi girma (20 μg/L), bi da bi. Misali, canje-canje a cikin DNA na wani nau'in na iya haifar da mummunan sakamako ga rayuwa ta ɗan gajeren lokaci da ta dogon lokaci, wanda ke haifar da raguwar yawan jama'a, canjin lafiyar haihuwa, haihuwa, asarar bambancin kwayoyin halitta, da canjin ƙimar ƙaura. Duk waɗannan abubuwan na iya shafar rayuwar nau'ikan halittu da kuma kula da su42,43. Samuwar rashin daidaituwar erythroid na iya nuna toshewar cytokines, wanda ke haifar da rarrabuwar ƙwayoyin halitta mara kyau (ƙwayoyin halitta masu kama da juna)44,45; ƙwayoyin halitta masu yawa suna fitowa daga membrane na nukiliya tare da lobes da yawa46, yayin da wasu rashin daidaituwar erythroid na iya alaƙa da haɓaka DNA, kamar kodan nukiliya/ƙwayoyin halitta47. Kasancewar ƙwayoyin halitta masu kama da juna na iya nuna rashin isashshen iskar oxygen, musamman a cikin ruwa mai gurɓata48,49. Apoptosis yana nuna mutuwar ƙwayoyin halitta50.
Wasu bincike sun kuma nuna tasirin kwayar halittar cypermethrin mai guba. Kabaña da abokan aikinsa 51 sun nuna kasancewar micronuclei da canje-canje na nukiliya kamar ƙwayoyin binucleated da ƙwayoyin apoptotic a cikin ƙwayoyin Odontophrynus americanus bayan fallasa su ga yawan cypermethrin (5000 da 10,000 μg L−1) na tsawon awanni 96. An kuma gano apoptosis da Cypermethrin ya haifar a cikin P. biligonigerus52 da Rhinella arenarum53. Waɗannan sakamakon sun nuna cewa cypermethrin yana da tasirin kwayar halitta mai guba akan nau'ikan halittu na ruwa kuma gwajin MN da ENA na iya zama alamar tasirin da ba a iya mutuwa ba ga halittu masu rai kuma yana iya aiki ga nau'ikan asali da kuma al'ummomin daji da ke fuskantar guba12.
Tsarin kasuwanci na cypermethrin yana haifar da babban haɗarin muhalli (mai tsanani da na yau da kullun), tare da manyan hedikwatocin da suka wuce matakin Hukumar Kare Muhalli ta Amurka (EPA) 54 wanda zai iya yin mummunan tasiri ga nau'in idan yana cikin muhalli. A cikin kimanta haɗarin da ke faruwa na yau da kullun, NOEC na mace-mace shine 3 μg L−1, yana tabbatar da cewa yawan da aka samu a cikin ruwa na iya haifar da haɗari ga nau'in55. NOEC mai kisa ga tsutsotsin R. arenarum da aka fallasa ga cakuda endosulfan da cypermethrin shine 500 μg L−1 bayan sa'o'i 168; wannan ƙimar ta ragu zuwa 0.0005 μg L−1 bayan sa'o'i 336. Marubutan sun nuna cewa tsawon lokacin da aka fallasa, ƙarancin yawan da ke cutar da nau'in. Hakanan yana da mahimmanci a nuna cewa ƙimar NOEC ta fi ta P. gracilis a lokaci guda, yana nuna cewa amsawar nau'in ga cypermethrin ta keɓance nau'in. Bugu da ƙari, dangane da mace-mace, ƙimar CHQ na P. gracilis bayan fallasa ga cypermethrin ya kai 64.67, wanda ya fi ƙimar da Hukumar Kare Muhalli ta Amurka ta gindaya54, kuma ƙimar CHQ na tsutsotsin R. arenarum shi ma ya fi wannan ƙimar (CHQ > 388.00 bayan sa'o'i 336), yana nuna cewa maganin kwari da aka yi nazari a kai yana da babban haɗari ga nau'ikan amphibian da yawa. Idan aka yi la'akari da cewa P. gracilis yana buƙatar kimanin kwanaki 30 don kammala metamorphosis56, za a iya kammala cewa yawan cypermethrin da aka yi nazari a kai na iya taimakawa wajen raguwar yawan jama'a ta hanyar hana mutanen da suka kamu da cutar shiga cikin manya ko matakin haihuwa a farkon shekarunsu.
A cikin kimanta haɗarin da aka ƙididdige na micronuclei da sauran matsalolin nukiliyar erythrocyte, ƙimar CHQ ta kasance daga 14.92 zuwa 97.00, wanda ke nuna cewa cypermethrin yana da yuwuwar haɗarin guba ga P. gracilis ko da a cikin mazauninsa na halitta. Idan aka yi la'akari da mace-mace, matsakaicin yawan mahaɗan xenobiotic da P. gracilis ke jurewa shine 4.24 μg L−1. Duk da haka, yawan da ya kai ƙasa da 1 μg/L shi ma ya nuna tasirin guba. Wannan gaskiyar na iya haifar da ƙaruwar adadin mutane marasa kyau57 kuma yana shafar ci gaba da haifuwar nau'ikan halittu a cikin mazauninsu, wanda ke haifar da raguwar yawan halittu masu rai.
Tsarin kasuwanci na cypermethrin na maganin kwari ya nuna babban guba mai tsanani da na yau da kullun ga P. gracilis. An lura da yawan mace-mace mai yawa, wataƙila saboda tasirin guba, kamar yadda aka tabbatar ta hanyar kasancewar ƙwayoyin micronuclei da erythrocyte nuclei, musamman ƙwayoyin serrated, ƙwayoyin lobed, da ƙwayoyin vesicular. Bugu da ƙari, nau'in da aka yi nazari a kai ya nuna ƙaruwar haɗarin muhalli, duka na tsanani da na yau da kullun. Waɗannan bayanai, tare da binciken da ƙungiyar bincikenmu ta yi a baya, sun nuna cewa har ma da nau'ikan cypermethrin na kasuwanci daban-daban har yanzu suna haifar da raguwar ayyukan acetylcholinesterase (AChE) da butyrylcholinesterase (BChE) da damuwa ta oxidative58, kuma sun haifar da canje-canje a ayyukan iyo da nakasa ta baki59 a cikin P. gracilis, wanda ke nuna cewa tsarin kasuwanci na cypermethrin yana da babban guba mai kisa da na ɗan lokaci ga wannan nau'in. Hartmann et al. 60 sun gano cewa tsarin kasuwanci na cypermethrin sune mafi guba ga P. gracilis da wani nau'in nau'in iri ɗaya (P. cuvieri) idan aka kwatanta da sauran magungunan kashe kwari guda tara. Wannan yana nuna cewa yawan sinadarin cypermethrin da aka amince da shi bisa doka don kare muhalli na iya haifar da yawan mace-mace da raguwar yawan jama'a na dogon lokaci.
Ana buƙatar ƙarin bincike don tantance gubar da maganin kwari ke yi wa dabbobin ruwa, domin yawan da ake samu a muhalli na iya haifar da mace-mace mai yawa kuma yana iya haifar da haɗari ga P. gracilis. Ya kamata a ƙarfafa bincike kan nau'ikan halittu masu ruwa-ruwa, domin bayanai kan waɗannan halittu ba su da yawa, musamman kan nau'ikan Brazil.
Gwajin guba na tsawon lokaci ya ɗauki tsawon awanni 168 (kwana 7) a ƙarƙashin yanayi mai tsauri kuma yawan gubar da ke cikin ƙasa shine: 1, 3, 6 da 20 μg ai L−1. A cikin gwaje-gwajen guda biyu, an kimanta tadpoles 10 a kowace rukunin magani tare da maimaitawa shida, jimlar tadpoles 60 a kowace taro. A halin yanzu, maganin da aka yi da ruwa kawai ya yi aiki azaman iko mara kyau. Kowace saitin gwaji ta ƙunshi farantin gilashi mai tsafta wanda ke da ƙarfin 500 ml da yawan tadpoles 1 a kowace ml 50 na bayani. An rufe kwalbar da fim ɗin polyethylene don hana ƙafewa kuma ana ci gaba da sanya iska ta shiga.
An yi nazarin ruwan ta hanyar sinadarai don tantance yawan magungunan kashe kwari a awanni 0, 96, da 168. A cewar Sabin et al. 68 da Martins et al. 69, an gudanar da binciken a dakin gwaje-gwajen nazarin magungunan kashe kwari (LARP) na Jami'ar Tarayya ta Santa Maria ta amfani da gas chromatography da aka haɗa zuwa triple quadrupole mass spectrometry (Varian model 1200, Palo Alto, California, Amurka). An nuna ƙimar tantance magungunan kashe kwari a cikin ruwa azaman ƙarin kayan aiki (Tebur SM1).
Don gwajin micronucleus (MNT) da gwajin rashin daidaituwar ƙwayoyin ja (RNA), an yi nazarin ƙwayoyin tadpoles 15 daga kowace ƙungiyar magani. An yi wa ƙwayoyin tadpoles maganin sa barci da 5% lidocaine (50 mg g-170) kuma an tattara samfuran jini ta hanyar huda zuciya ta amfani da sirinji mai heparinized da aka yarfa. An shirya ƙwayoyin jini a kan faifai masu ɓoye na microscope, an busar da su ta iska, an gyara su da 100% methanol (4 °C) na tsawon minti 2, sannan aka shafa musu maganin Giemsa 10% na tsawon minti 15 a cikin duhu. A ƙarshen aikin, an wanke zane-zanen da ruwan da aka tace don cire tabo mai yawa kuma an busar da su a zafin ɗaki.
An yi nazarin aƙalla RBCs 1000 daga kowace tadpole ta amfani da na'urar hangen nesa mai girman 100× tare da maƙasudi 71 don tantance kasancewar MN da ENA. An kimanta jimillar RBCs 75,796 daga tadpoles ta hanyar la'akari da yawan cypermethrin da sarrafawa. An yi nazarin gubar kwayoyin halitta bisa ga hanyar Carrasco et al. da Fenech et al.38,72 ta hanyar tantance yawan raunukan nukiliya masu zuwa: (1) ƙwayoyin anucleate: ƙwayoyin da ba su da ƙwayoyin cuta; (2) ƙwayoyin apoptotic: rarrabuwar nukiliya, mutuwar ƙwayoyin da aka tsara; (3) ƙwayoyin binucleate: ƙwayoyin da ke da ƙwayoyin cuta guda biyu; (4) ƙwayoyin nukiliya ko ƙwayoyin bleb: ƙwayoyin da ke da ƙwayoyin cuta tare da ƙananan fitowar membrane na nukiliya, blebs masu kama da girman micronuclei; (5) ƙwayoyin karyolyzed: ƙwayoyin da ke da tsarin tsakiya kawai ba tare da kayan ciki ba; (6) ƙwayoyin da aka yi wa ado: ƙwayoyin da ke da ƙwayoyin cuta tare da fasa ko ƙusoshi a bayyane a siffarsu, wanda kuma ake kira ƙwayoyin da ke da siffar koda; (7) ƙwayoyin da aka yi amfani da su wajen yin amfani da su: ƙwayoyin da suka fito daga nuclear sun fi girma fiye da vesicles da aka ambata a sama; da kuma (8) ƙwayoyin micro: ƙwayoyin da ke da ƙwayoyin da aka taƙaice da kuma raguwar cytoplasm. An kwatanta canje-canjen da sakamakon sarrafawa mara kyau.
An yi nazarin sakamakon gwajin guba mai tsanani (LC50) ta amfani da software na GBasic da kuma hanyar TSK-Trimmed Spearman-Karber74. An riga an gwada bayanan gwajin na yau da kullun don daidaiton kuskure (Shapiro-Wilks) da kuma daidaiton bambancin (Bartlett). An yi nazarin sakamakon ta amfani da nazarin bambancin hanya ɗaya (ANOVA). An yi amfani da gwajin Tukey don kwatanta bayanai a tsakaninsu, kuma an yi amfani da gwajin Dunnett don kwatanta bayanai tsakanin ƙungiyar magani da ƙungiyar kulawa mara kyau.
An yi nazarin bayanan LOEC da NOEC ta amfani da gwajin Dunnett. An yi gwaje-gwajen ƙididdiga ta amfani da software na Statistica 8.0 (StatSoft) tare da matakin mahimmanci na 95% (p < 0.05).
Lokacin Saƙo: Maris-13-2025