Kafukufukuyu adawunika zakupha, kuwopsa, komanso kuwopsa kwazamalondacypermetrinformulations kwa anuran tadpoles. Mu mayeso owopsa, kuchuluka kwa 100-800 μg/L kunayesedwa kwa 96 h. M'mayeso osatha, kuchuluka kwachilengedwe kwa cypermethrin (1, 3, 6, ndi 20 μg / L) adayesedwa kuti amwalire, kutsatiridwa ndi kuyezetsa kwa ma micronucleus ndi kusokonekera kwa ma cell ofiira amagazi kwa masiku 7. LC50 ya malonda a cypermethrin kupanga ma tadpoles anali 273.41 μg L-1. M'mayeso osatha, kuchuluka kwambiri (20 μg L-1) kudapangitsa kuti anthu azifa kwambiri kuposa 50%, chifukwa adapha theka la tadpoles omwe adayesedwa. Mayeso a micronucleus adawonetsa zotsatira zazikulu pa 6 ndi 20 μg L-1 ndi zolakwika zingapo za nyukiliya zinapezeka, kusonyeza kuti kupanga cypermethrin yamalonda ili ndi mphamvu ya genotoxic motsutsana ndi P. gracilis. Cypermethrin ndi chiopsezo chachikulu cha zamoyozi, zomwe zimasonyeza kuti zingayambitse mavuto ambiri komanso zimakhudza mphamvu za chilengedwechi panthawi yochepa komanso yayitali. Choncho, tinganene kuti mankhwala a cypermethrin ali ndi zotsatira zoopsa pa P. gracilis.
Chifukwa chakukula kosalekeza kwa ntchito zaulimi komanso kugwiritsa ntchito kwambirikuwononga tizirombomiyeso, nyama zam'madzi nthawi zambiri zimakumana ndi mankhwala ophera tizilombo1,2. Kuipitsa kwa madzi pafupi ndi minda yaulimi kungasokoneze chitukuko ndi moyo wa zamoyo zomwe sizinali zolinga monga amphibians.
Amphibians akukhala ofunika kwambiri pakuwunika matrices achilengedwe. Anurans amaonedwa kuti ndi bioindicators zabwino zoipitsa chilengedwe chifukwa cha makhalidwe awo apadera monga mkombero zovuta moyo, mofulumira mitengo mphutsi kukula, trophic udindo, permeable skin10,11, kudalira madzi kwa kubalana12 ndi mazira osatetezedwa11,13,14. Chule chaching'ono cham'madzi (Physalaemus gracilis), chomwe chimadziwika kuti kulira, chawonetsedwa kuti ndi mtundu wa bioindicator wowononga mankhwala ophera tizilombo4,5,6,7,15. Mitunduyi imapezeka m'madzi oyimilira, malo otetezedwa kapena malo okhala ndi malo osiyanasiyana ku Argentina, Uruguay, Paraguay ndi Brazil1617 ndipo amaonedwa kuti ndi okhazikika ndi gulu la IUCN chifukwa cha kufalikira kwake ndi kulolerana kwa malo osiyanasiyana18.
Zotsatira za sublethal zafotokozedwa mu amphibians potsatira kukhudzana ndi cypermethrin, kuphatikizapo kusintha kwa khalidwe, morphological ndi biochemical mu tadpoles23,24,25, kusintha kwa imfa ndi nthawi ya metamorphosis, kusintha kwa enzymatic, kuchepa kwa kuphulika24,25, hyperactivity26, kuletsa kwa cholinesterase7 ntchito ndi kusintha28m28. Komabe, maphunziro a genotoxic zotsatira za cypermethrin mu amphibians ndi ochepa. Chifukwa chake, ndikofunikira kuwunika kuthekera kwa mitundu ya anuran ku cypermethrin.
Kuwonongeka kwa chilengedwe kumakhudza kukula ndi chitukuko cha amphibians, koma choopsa kwambiri ndi kuwonongeka kwa majini kwa DNA chifukwa cha mankhwala ophera tizilombo13. Kusanthula kwa ma cell a magazi ndi chizindikiro chofunikira kwambiri cha kuipitsidwa ndi kuopsa kwa chinthu ku mitundu yakuthengo29. Kuyesa kwa micronucleus ndi imodzi mwa njira zomwe zimagwiritsidwa ntchito kwambiri pozindikira kuopsa kwa mankhwala m'chilengedwe30. Ndi njira yofulumira, yothandiza komanso yotsika mtengo yomwe ili chisonyezero chabwino cha kuipitsidwa kwa mankhwala a zamoyo monga amphibians31,32 ndipo ikhoza kupereka chidziwitso chokhudzana ndi zowononga zowonongeka za genotoxic33.
Cholinga cha kafukufukuyu chinali kuyesa kuthekera kwapoizoni kwa ma cypermethrin opangira malonda a tadpoles ang'onoang'ono am'madzi pogwiritsa ntchito mayeso a micronucleus komanso kuwunika kwachilengedwe.
Kufa kochulukirachulukira (%) kwa P. gracilis tadpoles komwe kumawonekera kumagulu osiyanasiyana amalonda a cypermethrin panthawi yovuta ya mayeso.
Kufa kochulukirachulukira (%) kwa P. gracilis tadpoles komwe kumawonekera kumagulu osiyanasiyana amalonda a cypermethrin panthawi ya mayeso osatha.
Kufa kwakukulu komwe kumawonedwa kudachitika chifukwa cha zotsatira za genotoxic mu amphibians omwe amapezeka mumitundu yosiyanasiyana ya cypermethrin (6 ndi 20 μg/L), monga umboni wa kukhalapo kwa ma micronuclei (MN) ndi zolakwika za nyukiliya mu erythrocytes. Kupanga kwa MN kumasonyeza zolakwika mu mitosis ndipo zimagwirizanitsidwa ndi kusamangika kosauka kwa ma chromosome ku ma microtubules, kuwonongeka kwa mapuloteni omwe amachititsa kuti chromosome itengeke ndi kunyamula, zolakwika za kusiyana kwa chromosome ndi zolakwika mu kukonza kuwonongeka kwa DNA38,39 ndipo zikhoza kukhala zokhudzana ndi mankhwala ophera tizilombo oxidative stress40,41. Zolakwika zina zidawonedwa pazowunikira zonse zomwe zidawunikidwa. Kuchulukirachulukira kwa cypermethrin kumawonjezera zovuta za nyukiliya mu erythrocytes ndi 5% ndi 20% pamlingo wotsika kwambiri (1 μg/L) ndi mlingo wapamwamba kwambiri (20 μg/L), motsatana. Mwachitsanzo, kusintha kwa DNA ya zamoyo kumatha kukhala ndi zotsatirapo zoyipa pakukhala ndi moyo kwakanthawi kochepa komanso kwanthawi yayitali, zomwe zimapangitsa kuchepa kwa chiwerengero cha anthu, kusintha nyonga yobereka, kubereketsa, kutayika kwa mitundu yosiyanasiyana ya majini, ndi kusintha kwa kusamuka. Zinthu zonsezi zimatha kukhudza kupulumuka kwa zamoyo ndi kukonza42,43. Kupanga zolakwika za erythroid kungasonyeze chipika mu cytokinesis, zomwe zimapangitsa kuti maselo agawanika (binucleated erythrocytes) 44,45; ma nuclei ambiri ndi ma protrusions a nyukiliya okhala ndi ma lobes46 angapo, pomwe zovuta zina za erythroid zitha kulumikizidwa ndi kukulitsa kwa DNA, monga impso za nyukiliya / blebs47. Kukhalapo kwa anucleated erythrocytes kungasonyeze kusokonezeka kwa kayendedwe ka mpweya, makamaka m'madzi oipitsidwa48,49. Apoptosis imasonyeza kufa kwa maselo50.
Kafukufuku wina wawonetsanso zotsatira za genotoxic za cypermethrin. Kabaña et al.51 adawonetsa kukhalapo kwa micronuclei ndi kusintha kwa nyukiliya monga ma cell a binucleated ndi ma cell apoptotic m'maselo a Odontophrynus americanus atatha kuwonetsa kwambiri cypermethrin (5000 ndi 10,000 μg L-1) kwa 96 h. Cypermethrin-induced apoptosis inapezekanso mu P. biligonigerus52 ndi Rhinella arenarum53. Zotsatirazi zikusonyeza kuti cypermethrin ili ndi zotsatira za genotoxic pamitundu yambiri ya zamoyo zam'madzi komanso kuti kuyesa kwa MN ndi ENA kungakhale chizindikiro cha zotsatira zowonongeka kwa amphibians ndipo zingakhale zogwira ntchito kwa zamoyo zamtundu ndi zakutchire zomwe zimakhudzidwa ndi toxicants12.
Mapangidwe a zamalonda a cypermethrin amabweretsa chiwopsezo chachikulu cha chilengedwe (chowopsa komanso chosatha), pomwe ma HQ amatha kupitilira mulingo wa54 wa US Environmental Protection Agency (EPA) zomwe zitha kusokoneza zamoyo ngati zilipo m'chilengedwe. Pakuwunika kwachiwopsezo chosatha, NOEC yakufa inali 3 μg L-1, kutsimikizira kuti kuchuluka komwe kumapezeka m'madzi kumatha kukhala pachiwopsezo kwa mitundu55. NOEC yakupha ya R. arenarum mphutsi zowonekera ku chisakanizo cha endosulfan ndi cypermethrin chinali 500 μg L-1 pambuyo pa 168 h; mtengo uwu unatsikira ku 0.0005 μg L-1 pambuyo pa 336 h. Olembawo akuwonetsa kuti nthawi yayitali kuwonekera, kumachepetsa kwambiri zomwe zimakhala zovulaza kwa zamoyo. Ndikofunikiranso kuwunikira kuti mikhalidwe ya NOEC yadali yapamwamba kuposa ya P. gracilis panthawi yomweyi, kuwonetsa kuti mitundu yamitundu ya cypermethrin ndiyosiyana ndi mitundu. Kuwonjezera apo, ponena za imfa, mtengo wa CHQ wa P. gracilis pambuyo powonekera kwa cypermethrin unafika pa 64.67, womwe ndi wapamwamba kuposa mtengo wamtengo wapatali womwe unakhazikitsidwa ndi US Environmental Protection Agency54, ndipo mtengo wa CHQ wa R. arenarum mphutsi unalinso wapamwamba kusiyana ndi mtengo uwu (CHQ> 388.00 388.00h pambuyo pa 338. mitundu ingapo ya amphibians. Poganizira kuti P. gracilis imafuna pafupifupi masiku 30 kuti amalize metamorphosis56, tinganene kuti kuchuluka kwa cypermethrin komwe kumaphunziridwa kungathandize kuchepetsa chiwerengero cha anthu poletsa anthu omwe ali ndi kachilombo kuti alowe m'magulu akuluakulu kapena obereketsa adakali aang'ono.
Powerengera chiopsezo cha micronuclei ndi zovuta zina za nyukiliya za erythrocyte, miyeso ya CHQ idachokera pa 14.92 mpaka 97.00, zomwe zikuwonetsa kuti cypermethrin inali ndi chiopsezo cha genotoxic kwa P. gracilis ngakhale m'malo ake achilengedwe. Poganizira za imfa, kuchuluka kwa mankhwala a xenobiotic omwe angaloledwe ndi P. gracilis anali 4.24 μg L-1. Komabe, ndende zotsika ngati 1 μg/L zidawonetsanso zotsatira za genotoxic. Izi zingapangitse kuti chiwerengero cha anthu omwe ali ndi matenda achuluke57 komanso kusokoneza chitukuko ndi kuberekana kwa zamoyo m'madera awo, zomwe zimapangitsa kuti chiwerengero cha amphibian chichepe.
Mapangidwe amalonda a mankhwala ophera tizilombo a cypermethrin adawonetsa kuopsa koopsa komanso kosatha kwa P. gracilis. Kupha anthu ambiri kunawonedwa, mwinamwake chifukwa cha zotsatira za poizoni, monga umboni wa kukhalapo kwa micronuclei ndi erythrocyte nyukiliya zolakwika, makamaka serrated nuclei, lobed nuclei, ndi vesicular nuclei. Kuphatikiza apo, mitundu yomwe idaphunziridwayo idawonetsa kuwopsa kwachilengedwe, komwe kumakhala kowopsa komanso kosatha. Deta iyi, pamodzi ndi kafukufuku wam'mbuyomu ndi gulu lathu lofufuza, zikuwonetsa kuti ngakhale mitundu yosiyanasiyana yazamalonda ya cypermethrin idapangitsabe kuchepa kwa acetylcholinesterase (AChE) ndi butyrylcholinesterase (BChE) ndi oxidative stress58, ndipo zidapangitsa kusintha kwa ntchito yosambira ndi zofooka zapakamwa59 mu P. gracilimeth formulations of lelethal subletter and lelethang letharing kawopsedwe kwa mitundu iyi. Hartmann et al. 60 anapeza kuti malonda a cypermethrin anali oopsa kwambiri kwa P. gracilis ndi mitundu ina ya mtundu womwewo (P. cuvieri) poyerekeza ndi mankhwala ena asanu ndi anayi. Izi zikusonyeza kuti cypermethrin yovomerezeka mwalamulo pofuna kuteteza chilengedwe kungayambitse imfa zambiri komanso kuchepa kwa anthu kwa nthawi yaitali.
Maphunziro owonjezera akufunika kuti awone momwe mankhwala ophera tizilombo amadziwira kwa amphibians, chifukwa kuchuluka komwe kumapezeka m'chilengedwe kungayambitse kufa kwakukulu ndikuyika chiopsezo kwa P. gracilis. Kafukufuku wokhudza zamoyo zam'mlengalenga ayenera kulimbikitsidwa, chifukwa zambiri za zamoyozi ndizosowa, makamaka zamoyo zaku Brazil.
Kuyesa kwapoizoni kosatha kunachitika kwa 168 h (masiku 7) pansi pazikhalidwe zosasunthika ndipo kuchuluka kwake kunali: 1, 3, 6 ndi 20 μg ai L-1. Pazoyeserera zonse ziwiri, ma tadpoles 10 pa gulu lililonse lamankhwala adawunikidwa ndi zotsatsira zisanu ndi chimodzi, pazambiri za 60 tadpoles pagulu lililonse. Panthawiyi, mankhwala a madzi okha anali ngati kulamulira koipa. Kukonzekera kulikonse kunali ndi mbale yagalasi yosabala yokhala ndi mphamvu ya 500 ml ndi kachulukidwe ka tadpole 1 pa 50 ml ya yankho. Botololo linali lophimbidwa ndi filimu ya polyethylene kuti lisatuluke nthunzi ndipo limapitilizidwa ndi mpweya.
Madziwo adawunikidwa ndi mankhwala kuti adziwe kuchuluka kwa mankhwala ophera tizilombo pa 0, 96, ndi 168 h. Malinga ndi Sabin et al. 68 ndi Martins et al. 69 , kusanthula kunachitika ku Pesticide Analysis Laboratory (LARP) ya Federal University of Santa Maria pogwiritsa ntchito chromatography ya gas yophatikizidwa ndi katatu quadrupole mass spectrometry (Varian model 1200, Palo Alto, California, USA). Kutsimikiza kwa kuchuluka kwa mankhwala ophera tizilombo m'madzi kumawonetsedwa ngati zowonjezera (Table SM1).
Pa mayeso a micronucleus (MNT) ndi kuyesa kwa nyukiliya yofiira (RNA), ma tadpole 15 ochokera ku gulu lililonse lamankhwala adawunikidwa. Tadpoles anali anesthetized ndi 5% lidocaine wa (50 mg g-170) ndi zitsanzo magazi anatengedwa ndi mtima puncture ntchito disposable heparinized syringe. Magazi opaka magazi adakonzedwa pazithunzi za microscope wosabala, zouma mpweya, zokhazikika ndi 100% methanol (4 ° C) kwa 2 min, ndiyeno amadetsedwa ndi 10% yankho la Giemsa kwa mphindi 15 mumdima. Kumapeto kwa ndondomekoyi, slide amatsukidwa ndi madzi osungunuka kuti achotse banga lochulukirapo ndikuumitsa kutentha.
Pafupifupi ma RBC a 1000 kuchokera ku tadpole iliyonse adawunikidwa pogwiritsa ntchito maikulosikopu ya 100 × ndi cholinga cha 71 kuti adziwe kupezeka kwa MN ndi ENA. Ma RBC okwana 75,796 ochokera ku tadpoles adawunikidwa poganizira kuchuluka kwa cypermethrin ndi kuwongolera. Genotoxicity idawunikidwa molingana ndi njira ya Carrasco et al. ndi Fenech et al.38,72 pozindikira kuchuluka kwa zilonda za nyukiliya zotsatirazi: (1) maselo a anucleate: maselo opanda ma nuclei; (2) ma cell a apoptotic: kugawikana kwa nyukiliya, kufa kwadongosolo; (3) ma cell a binucleate: maselo okhala ndi ma nuclei awiri; (4) masamba a nyukiliya kapena ma cell a bleb: maselo okhala ndi ma nuclei okhala ndi tinthu tating'onoting'ono ta nyukiliya, ma blebs ofanana kukula kwa ma micronuclei; (5) maselo a karyolyzed: maselo okhala ndi ndondomeko yokha ya phata popanda zinthu zamkati; (6) ma cell notched: maselo okhala ndi ma nuclei okhala ndi ming'alu yoonekeratu kapena notches mu mawonekedwe awo, amatchedwanso nyukiliya yooneka ngati impso; (7) maselo a lobulated: maselo okhala ndi zida za nyukiliya zazikulu kuposa ma vesicles omwe tawatchulawa; ndi (8) ma microcell: maselo okhala ndi nyukiliya yofupikitsidwa ndi cytoplasm yocheperako. Zosinthazo zidafaniziridwa ndi zotsatira zoyipa zowongolera.
Zotsatira za acute toxicity test (LC50) zidawunikidwa pogwiritsa ntchito pulogalamu ya GBasic ndi TSK-Trimmed Spearman-Karber method74. Deta yoyezetsa yosatha idayesedwa kale kuti ikhale yolakwika (Shapiro-Wilks) ndi homogeneity of variance (Bartlett). Zotsatirazo zidawunikidwa pogwiritsa ntchito njira imodzi yowunikira kusiyana (ANOVA). Kuyesa kwa Tukey kunagwiritsidwa ntchito kuyerekeza deta pakati pawo, ndipo kuyesa kwa Dunnett kunagwiritsidwa ntchito kuyerekeza deta pakati pa gulu lachipatala ndi gulu lolamulira loipa.
Zambiri za LOEC ndi NOEC zidawunikidwa pogwiritsa ntchito mayeso a Dunnett. Mayeso owerengera adachitidwa pogwiritsa ntchito pulogalamu ya Statistica 8.0 (StatSoft) yokhala ndi gawo lofunikira la 95% (p <0.05).
Nthawi yotumiza: Mar-13-2025