Amfani da magungunan kashe kwari na roba ya haifar da matsaloli da dama, ciki har da fitowar kwayoyin halitta masu jure wa cututtuka, lalacewar muhalli da kuma cutar da lafiyar dan adam. Saboda haka, sabbin ƙwayoyin cutamagungunan kashe kwariAna buƙatar gaggawa don tabbatar da cewa ƙwayoyin cuta masu lafiya ga lafiyar ɗan adam da muhalli suna da aminci. A cikin wannan binciken, an yi amfani da sinadarin rhamnolipid biosurfactant da Enterobacter cloacae SJ2 ya samar don tantance guba ga tsutsotsin sauro (Culex quinquefasciatus) da termite (Odontotermes obesus). Sakamakon ya nuna cewa akwai adadin mace-mace da ya dogara da kashi tsakanin jiyya. An ƙayyade ƙimar LC50 (50% yawan mutuwa) a cikin awanni 48 ga tsutsotsi da tsutsotsi na tsutsotsi ta amfani da hanyar daidaita lanƙwasa mara layi. Sakamakon ya nuna cewa ƙimar LC50 na awanni 48 (95% tazara amincewa) na aikin tsutsotsi da hana tururuwa na biosurfactant sune 26.49 mg/L (tsakanin 25.40 zuwa 27.57) da 33.43 mg/L (tsakanin 31.09 zuwa 35.68) bi da bi. A cewar binciken histopathological, magani da biosurfactants ya haifar da mummunan lalacewa ga kyallen organelle na tsutsotsi da tururuwa. Sakamakon wannan binciken ya nuna cewa kwayoyin halitta masu rai da Enterobacter cloacae SJ2 ke samarwa kayan aiki ne mai kyau kuma mai yuwuwa mai tasiri don sarrafa Cx. quinquefasciatus da O. obesus.
Kasashen da ke fama da cututtuka masu yawa da sauro ke yadawa sun yadu. Mutane sama da 400,000 ne ke mutuwa sakamakon zazzabin cizon sauro kowace shekara, kuma wasu manyan biranen suna fuskantar annoba ta cututtuka masu tsanani kamar dengue, yellow fever, chikungunya da Zika.2 Cututtukan da ake kamuwa da su ta hanyar vector suna da alaƙa da kamuwa da cuta ɗaya cikin shida a duniya, inda sauro ke haifar da mafi yawan lokuta3,4. Culex, Anopheles da Aedes su ne nau'ikan sauro uku da aka fi dangantawa da yaɗuwar cututtuka5. Yaɗuwar zazzabin cizon sauro, kamuwa da cuta da sauro na Aedes aegypti ke yadawa, ya ƙaru a cikin shekaru goma da suka gabata kuma yana haifar da babbar barazana ga lafiyar jama'a4,7,8. A cewar Hukumar Lafiya ta Duniya (WHO), sama da kashi 40% na al'ummar duniya suna cikin haɗarin kamuwa da zazzabin cizon sauro, inda sabbin mutane miliyan 50-100 ke kamuwa da cutar kowace shekara a cikin ƙasashe sama da 1009,10,11. Zazzabin cizon sauro ya zama babbar matsala ga lafiyar jama'a yayin da yawan kamuwa da cutar ya ƙaru a duk duniya12,13,14. Anopheles gambiae, wanda aka fi sani da sauro na Afirka Anopheles, shine mafi mahimmancin hanyar kamuwa da cutar zazzabin cizon sauro na ɗan adam a yankuna masu zafi da kuma ƙasa da wurare masu zafi15. Kwayar cutar West Nile, St. Louis encephalitis, Japan encephalitis, da cututtukan ƙwayoyin cuta na dawakai da tsuntsaye ana ɗaukar su ta hanyar sauro na Culex, wanda galibi ake kira sauro na gida. Bugu da ƙari, suna kuma ɗauke da cututtukan ƙwayoyin cuta da na kwayan cuta16. Akwai nau'ikan tururuwa sama da 3,000 a duniya, kuma sun kasance a duniya sama da shekaru miliyan 15017. Yawancin kwari suna rayuwa a cikin ƙasa kuma suna cin kayan itace da itace waɗanda ke ɗauke da cellulose. Tururuwan Indiya Odontotermes obesus wani muhimmin kwari ne wanda ke haifar da mummunar lalacewa ga amfanin gona da bishiyoyi masu mahimmanci18. A yankunan noma, kamuwa da tururuwa a matakai daban-daban na iya haifar da mummunar lalacewar tattalin arziki ga amfanin gona daban-daban, nau'ikan bishiyoyi da kayan gini. Tururuwa kuma na iya haifar da matsalolin lafiyar ɗan adam19.
Batun juriya daga ƙananan halittu da kwari a fannin magunguna da noma na yau yana da sarkakiya20,21. Saboda haka, kamfanonin biyu ya kamata su nemi sabbin magungunan kashe ƙwayoyin cuta masu inganci da kuma magungunan kashe ƙwayoyin cuta masu aminci. Ana samun magungunan kashe ƙwayoyin cuta na roba yanzu kuma an nuna cewa suna da saurin yaɗuwa kuma suna korar kwari marasa amfani22. A cikin 'yan shekarun nan, bincike kan ƙwayoyin cuta ya faɗaɗa saboda amfaninsu a masana'antu daban-daban. Kwayoyin halitta suna da matuƙar amfani kuma suna da matuƙar muhimmanci a fannin noma, gyaran ƙasa, fitar da mai, ƙwayoyin cuta da cire kwari, da sarrafa abinci23,24. Kwayoyin halitta ko ƙwayoyin cuta surfactants sinadarai ne na halittu da ƙwayoyin cuta ke samarwa kamar ƙwayoyin cuta, yisti da fungi a wuraren da ke bakin teku da wuraren da aka gurbata mai25,26. Kwayoyin halitta da ƙwayoyin halitta da aka samo daga sinadarai nau'i biyu ne da ake samu kai tsaye daga muhallin halitta27. Ana samun nau'ikan halittu daban-daban daga wuraren da ke cikin ruwa28,29. Saboda haka, masana kimiyya suna neman sabbin fasahohi don samar da halittu masu rai bisa ga ƙwayoyin halitta30,31. Ci gaban da aka samu a irin wannan bincike ya nuna muhimmancin waɗannan mahaɗan halittu don kare muhalli32. Bacillus, Pseudomonas, Rhodococcus, Alcaligenes, Corynebacterium da waɗannan ƙwayoyin cuta wakilai ne da aka yi nazari sosai a kansu23,33.
Akwai nau'ikan halittu masu rai da yawa waɗanda ke da nau'ikan amfani iri-iri34. Babban fa'idar waɗannan mahaɗan shine cewa wasu daga cikinsu suna da ayyukan kashe ƙwayoyin cuta, kashe ƙwari da kuma kashe kwari. Wannan yana nufin cewa ana iya amfani da su a masana'antar noma, sinadarai, magunguna da kwalliya35,36,37,38. Saboda halittu masu rai gabaɗaya suna da lalacewa kuma suna da amfani ga muhalli, ana amfani da su a cikin shirye-shiryen kula da kwari da aka haɗa don kare amfanin gona39. Don haka, an sami ilimin asali game da ayyukan tsutsa da hana tururuwa na ƙwayoyin cuta masu rai da Enterobacter cloacae SJ2 ke samarwa. Mun binciki mace-mace da canje-canje na histological lokacin da aka fallasa su ga yawan ƙwayoyin halitta masu rai da rhamnolipid daban-daban. Bugu da ƙari, mun kimanta shirin kwamfuta na Tsarin Tsarin-Ayyukan Kima (QSAR) da ake amfani da shi sosai don tantance guba mai tsanani ga ƙananan algae, daphnia, da kifi.
A cikin wannan binciken, an gwada aikin antitermite (guba) na sinadaran halittu masu tsabta a cikin adadi daban-daban tun daga 30 zuwa 50 mg/ml (a tazara 5 mg/ml) akan tururuwan Indiya, O. obesus da nau'in na huɗu) Kimanta. Tsutsotsin instar Cx. Tsutsotsin sauro quinquefasciatus. Yawan Biosurfactant LC50 a cikin awanni 48 akan O. obesus da Cx. C. solanacearum. An gano tsutsotsin sauro ta amfani da hanyar daidaita lanƙwasa mara layi. Sakamakon ya nuna cewa mace-macen tururuwa ya ƙaru tare da ƙaruwar yawan biosurfactant. Sakamakon ya nuna cewa biosurfactant yana da aikin kashe ƙwari (Hoto na 1) da kuma aikin hana tururuwa (Hoto na 2), tare da ƙimar LC50 na awanni 48 (95% CI) na 26.49 mg/L (25.40 zuwa 27.57) da 33.43 mg/l (Hoto na 31.09 zuwa 35.68), bi da bi (Tebur na 1). Dangane da guba mai tsanani (awanni 48), an rarraba biosurfactant a matsayin "mai cutarwa" ga ƙwayoyin da aka gwada. Biosurfactant da aka samar a cikin wannan binciken ya nuna kyakkyawan aikin kashe ƙwari tare da mace-mace 100% cikin awanni 24-48 na fallasa.
Lissafa ƙimar LC50 don aikin kashe ƙurji. Daidaita lanƙwasa mai lanƙwasa mara layi (layi mai ƙarfi) da tazara amincewa 95% (yanki mai inuwa) don mace-macen dangi (%).
Lissafa ƙimar LC50 don ayyukan hana tururuwa. Daidaita lanƙwasa mai lanƙwasa mara layi (layi mai ƙarfi) da tazara amincewa 95% (yanki mai inuwa) don mace-mace mai dangantaka (%).
A ƙarshen gwajin, an lura da canje-canjen siffofi da rashin daidaituwa a ƙarƙashin na'urar hangen nesa. An lura da canje-canjen siffofi a cikin ƙungiyoyin sarrafawa da waɗanda aka yi wa magani a girman sau 40. Kamar yadda aka nuna a Hoto na 3, raunin girma ya faru a yawancin tsutsotsi da aka yi wa magani da biosurfactants. Hoto na 3a yana nuna Cx. quinquefasciatus na yau da kullun, Hoto na 3b yana nuna Cx mara kyau. Yana haifar da tsutsotsi na nematode guda biyar.
Tasirin alluran biosurfactants na ƙarƙashin mutuwa (LC50) akan ci gaban tsutsotsin Culex quinquefasciatus. Hoton haske na microscopy (a) na Cx na yau da kullun a girman 40×. quinquefasciatus (b) Cx mara kyau. Yana haifar da tsutsotsin nematode guda biyar.
A cikin wannan binciken, binciken histological na tsutsotsi da aka yi wa magani (Hoto na 4) da tururuwa (Hoto na 5) ya nuna wasu matsaloli da dama, ciki har da raguwar yankin ciki da lalacewar tsokoki, yadudduka na epithelial da fata. tsakiyar hanji. Histology ya bayyana hanyar da ake amfani da ita wajen hana aikin biosurfactant da aka yi amfani da shi a cikin wannan binciken.
Tarihin cututtukan ƙwayoyin cuta na ƙwai na Cx na 4th instar da ba a yi musu magani ba. tsutsotsin quinquefasciatus (ikon sarrafawa: (a,b)) kuma an yi musu magani da biosurfactant (magani: (c,d)). Kibiyoyi suna nuna epithelium na hanji da aka yi wa magani (epi), nuclei (n), da tsoka (mu). Bar = 50 µm.
Histopathology na O. obesus na yau da kullun wanda ba a yi masa magani ba (ikon sarrafawa: (a,b)) da biosurfactant da aka yi wa magani (magani: (c,d)). Kibiyoyi suna nuna epithelium na hanji (epi) da tsoka (mu), bi da bi. Bar = 50 µm.
A cikin wannan binciken, an yi amfani da ECOSAR don annabta guba mai tsanani na samfuran rhamnolipid biosurfactant ga masu samar da farko (kore algae), masu amfani da farko (ƙudan ruwa) da masu amfani da sakandare (kifi). Wannan shirin yana amfani da samfuran hadaddun tsari-aiki mai zurfi don kimanta guba bisa ga tsarin kwayoyin halitta. Tsarin yana amfani da software na aiki-tsari (SAR) don ƙididdige guba mai tsanani da na dogon lokaci na abubuwa ga nau'ikan ruwa. Musamman, Tebur na 2 ya taƙaita matsakaicin yawan mutuwa (LC50) da matsakaicin yawan tasiri (EC50) ga nau'ikan da yawa. An rarraba wanda ake zargi da guba zuwa matakai huɗu ta amfani da Tsarin Rarrabawa da Lakabi na Sinadarai na Duniya (Tebur 3).
Kula da cututtukan da ke ɗauke da ƙwayoyin cuta, musamman nau'ikan sauro da sauro na Aedes. Masarawa, yanzu suna aiki tuƙuru 40,41,42,43,44,45,46. Duk da cewa wasu magungunan kashe kwari da ake samu a sinadarai, kamar pyrethroids da organophosphates, suna da ɗan amfani, suna haifar da manyan haɗari ga lafiyar ɗan adam, gami da ciwon suga, matsalolin haihuwa, matsalolin jijiyoyi, ciwon daji, da cututtukan numfashi. Bugu da ƙari, bayan lokaci, waɗannan kwari na iya zama masu jure musu13,43,48. Don haka, ingantattun matakan kula da halittu masu kyau da kuma waɗanda ba su da muhalli za su zama hanyar da ta fi shahara ta maganin sauro49,50. Benelli51 ya ba da shawarar cewa kula da ƙwayoyin sauro da wuri zai fi tasiri a yankunan birane, amma ba su ba da shawarar amfani da tsutsotsi a yankunan karkara52 ba. Tom et al 53 sun kuma ba da shawarar cewa sarrafa sauro a matakan da ba su kai ba zai zama dabara mai aminci da sauƙi saboda sun fi saurin kamuwa da cututtukan da ke shafar sarrafa sauro54.
Samar da sinadaran biosurfactant ta hanyar wani nau'in halitta mai ƙarfi (Enterobacter cloacae SJ2) ya nuna inganci mai kyau da kuma mai kyau. Bincikenmu na baya ya ruwaito cewa Enterobacter cloacae SJ2 yana inganta samar da sinadaran biosurfactant ta amfani da sigogin physicochemical26. A cewar bincikensu, mafi kyawun yanayi don samar da sinadaran biosurfactant ta hanyar E. cloacae isolate mai yuwuwa shine haɗuwa na tsawon awanni 36, tashin hankali a 150 rpm, pH 7.5, 37 °C, gishiri 1 ppt, 2% glucose a matsayin tushen carbon, 1% yisti. An yi amfani da sinadarin a matsayin tushen nitrogen don samun 2.61 g/L biosurfactant. Bugu da ƙari, an siffanta biosurfactants ta amfani da TLC, FTIR da MALDI-TOF-MS. Wannan ya tabbatar da cewa rhamnolipid biosurfactant ne. Glycosipid biosurfactants sune aji mafi zurfi da aka yi nazari a kai na sauran nau'ikan biosurfactants55. Sun ƙunshi sassan carbohydrate da lipid, galibi sarƙoƙi na fatty acid. Daga cikin glycolipids, manyan wakilai sune rhamnolipid da sophorolipid56. Rhamnolipids sun ƙunshi sassan rhamnose guda biyu da ke da alaƙa da mono‐ ko di‐β‐hydroxydecanoic acid57. Amfani da rhamnolipids a masana'antun likitanci da magunguna an tabbatar da su sosai, baya ga amfani da su kwanan nan a matsayin magungunan kashe kwari59.
Hulɗar biosurfactant da yankin hydrophobic na numfashi yana ba da damar ruwa ya ratsa cikin ramin ciki, ta haka yana ƙara hulɗar tsutsotsi da muhallin ruwa. Kasancewar biosurfactants kuma yana shafar trachea, wanda tsawonsa yake kusa da saman, wanda ke sauƙaƙa tsutsotsi su rarrafe zuwa saman su yi numfashi. Sakamakon haka, matsin lamba na saman ruwa yana raguwa. Tunda tsutsotsi ba za su iya haɗuwa da saman ruwan ba, suna faɗuwa zuwa ƙasan tanki, suna kawo cikas ga matsin lamba na hydrostatic, wanda ke haifar da kashe kuzari da yawa da mutuwa ta hanyar nutsewa38,60. Ghribi61 ya sami irin wannan sakamakon, inda wani biosurfactant da Bacillus subtilis ya samar ya nuna aikin larvicidal akan Ephestia kuehniella. Hakazalika, aikin larvicidal na Cx. Das da Mukherjee23 suma sun tantance tasirin lipopeptides na cyclic akan tsutsotsin quinquefasciatus.
Sakamakon wannan binciken ya shafi aikin kashe ƙurji na ƙwayoyin halittar rhamnolipid akan Cx. Kashe sauro na quinquefasciatus ya yi daidai da sakamakon da aka buga a baya. Misali, ana amfani da ƙwayoyin halittar da ke tushen surfactin waɗanda ƙwayoyin cuta daban-daban na halittar Bacillus ke samarwa. kuma Pseudomonas spp. Wasu rahotanni na farko64,65,66 sun ba da rahoton aikin kashe ƙurji na ƙwayoyin halittar lipopeptide daga Bacillus subtilis23. Deepali et al. 63 sun gano cewa ƙwayoyin halittar rhamnolipid da aka ware daga Stenotropomonas maltophilia suna da ƙarfin aikin kashe ƙurji a yawan 10 mg/L. Silva et al. 67 sun ba da rahoton aikin kashe ƙurji na ƙwayoyin halittar rhamnolipid akan Ae a yawan 1 g/L. Aedes aegypti. Kanakdande et al. 68 sun ruwaito cewa ƙwayoyin halitta masu ɗauke da sinadarin lipopeptide da Bacillus subtilis ke samarwa sun haifar da mace-mace gaba ɗaya a cikin tsutsotsin Culex da tururuwa tare da ɓangaren lipophilic na Eucalyptus. Hakazalika, Masendra et al. 69 sun ba da rahoton mutuwar tururuwa ma'aikata (Cryptotermes cynocephalus Light.) na kashi 61.7% a cikin ɓangaren lipophilic n-hexane da EtOAc na ɗanyen E..
Parthipan da abokan aikinsa 70 sun ba da rahoton amfani da magungunan kashe kwari na lipopeptide biosurfactants da Bacillus subtilis A1 da Pseudomonas stutzeri NA3 suka samar a kan Anopheles Stephensi, wani nau'in kwayar cutar malaria mai suna Plasmodium. Sun lura cewa tsutsotsi da jaki sun rayu tsawon lokaci, suna da gajerun lokutan oviposition, ba su da tsabta, kuma suna da gajerun tsawon rai idan aka yi musu magani da yawan biosurfactants daban-daban. Ƙimar LC50 da aka lura da ita na B. subtilis biosurfactant A1 sun kasance 3.58, 4.92, 5.37, 7.10 da 7.99 mg/L ga yanayin tsutsotsi daban-daban (watau tsutsotsi I, II, III, IV da stage pupae) bi da bi. Idan aka kwatanta, biosurfactants na matakan tsutsotsi na I-IV da pupal na Pseudomonas stutzeri NA3 sun kasance 2.61, 3.68, 4.48, 5.55 da 6.99 mg/L, bi da bi. Ana kyautata zaton jinkirin da aka samu a yanayin tsutsotsi da 'yan jaki da suka tsira ya faru ne sakamakon manyan matsalolin da suka shafi jiki da kuma yadda kwayoyin cuta ke shafar su sakamakon maganin kwari71.
Nau'in Wickerhamomyces anomalus CCMA 0358 yana samar da biosurfactant mai aikin kashe ƙwari 100% akan sauro na Aedes. Tazara ta aegypti ta awanni 24 38 ta fi yadda Silva da abokan aikinsa suka ruwaito. An nuna cewa wani biosurfactant da aka samar daga Pseudomonas aeruginosa ta amfani da man sunflower a matsayin tushen carbon yana kashe 100% na tsutsotsi cikin awanni 48 67. Abinaya da abokan aikinsa 72 da Pradhan da abokan aikinsa 73 sun kuma nuna tasirin tsutsotsi ko kwari na surfactants da aka samar ta hanyar wasu nau'ikan Bacillus. Wani bincike da Senthil-Nathan da abokan aikinsa suka buga a baya ya gano cewa 100% na tsutsotsin sauro da aka fallasa ga tafkunan tsire-tsire za su iya mutuwa. 74.
Kimanta tasirin magungunan kwari masu kashe kwari a kan ilmin halittar kwari yana da matuƙar muhimmanci ga shirye-shiryen kula da kwari masu haɗaka domin allurai/mafi yawan ƙwayoyin cuta ba sa kashe kwari amma suna iya rage yawan ƙwayoyin cuta a cikin tsararraki masu zuwa ta hanyar lalata halayen halittu10. Siqueira da abokan aikinsa 75 sun lura da cikakken aikin kashe ƙwari (100% mace-mace) na rhamnolipid biosurfactant (300 mg/ml) lokacin da aka gwada su a adadi daban-daban tun daga 50 zuwa 300 mg/ml. Matakin tsutsotsi na nau'ikan Aedes aegypti. Sun yi nazarin tasirin lokaci zuwa mutuwa da yawan ƙwayoyin cuta masu kashe kwari akan rayuwar tsutsotsi da ayyukan iyo. Bugu da ƙari, sun lura da raguwar saurin iyo bayan awanni 24-48 na fallasa ga yawan ƙwayoyin cuta masu kashe kwari (misali, 50 mg/mL da 100 mg/mL). Ana tsammanin guba waɗanda ke da rawar da za su iya takawa wajen haifar da lalacewa da yawa ga kwari da aka fallasa76.
Binciken da aka yi a kan sakamakon bincikenmu ya nuna cewa kwayoyin halitta da Enterobacter cloacae SJ2 ke samarwa suna canza kyallen sauro (Cx. quinquefasciatus) da tsutsotsi (O. obesus). Ochola77 ta bayyana irin wannan rashin daidaituwar yanayi. Kamaraj da abokan aikinsa sun kuma bayyana irin wannan rashin daidaituwar yanayi a cikin An. tsutsotsin Stephanie sun fallasa su ga ƙwayoyin zinare. Vasantha-Srinivasan da abokan aikinsa sun kuma ba da rahoton cewa man fetur mai mahimmanci na jakar shepherd ya lalata ɗakin da kuma epithelial Layer na Aedes albopictus sosai. Aedes aegypti. Raghavendran da abokan aikinsa sun ba da rahoton cewa an yi wa tsutsotsin sauro magani da 500 mg/ml mycelial extract na wani naman gwari na Penicillium na gida. Ae suna nuna mummunan lalacewar histological. aegypti da Cx. Yawan mace-mace 80. A baya, an yi nazarin Abinaya da abokan aikinsa na huɗu na An. Stephensi da Ae. aegypti sun gano canje-canje da yawa a cikin Aedes aegypti da aka yi wa magani da B. licheniformis exopolysaccharides, gami da cecum na ciki, bugun tsoka, lalacewa da rashin tsari na ganglia na igiyar jijiya72. A cewar Raghavendran et al., bayan an yi wa maganin P. daleeae mycelial extract, ƙwayoyin tsakiyar hanji na sauro da aka gwada (ƙwayar cuta ta 4th instar) sun nuna kumburin lumen na hanji, raguwar abubuwan da ke cikin ƙwayoyin halitta, da kuma lalacewar nukiliya81. An lura da irin waɗannan canje-canjen a cikin tsutsar sauro da aka yi wa magani da ganyen echinacea, wanda ke nuna yuwuwar kashe kwari na mahaɗan da aka yi wa magani50.
Amfani da manhajar ECOSAR ya sami karbuwa a duniya baki daya82. Binciken da ake yi a yanzu ya nuna cewa guba mai tsanani da kwayoyin halitta na ECOSAR ke yi wa microalgae (C. vulgaris), kifi da ƙudaje na ruwa (D. magna) ke haifarwa ya fada cikin rukunin "guba" da Majalisar Dinkin Duniya ta ayyana83. Tsarin gubar muhalli na ECOSAR yana amfani da SAR da QSAR don annabta guba mai tsanani da na dogon lokaci na abubuwa kuma galibi ana amfani da shi don annabta gubar gurɓatattun abubuwa na halitta82,84.
An sayi Paraformaldehyde, sodium phosphate buffer (pH 7.4) da duk sauran sinadarai da aka yi amfani da su a wannan binciken daga HiMedia Laboratories, Indiya.
An gudanar da samar da biosurfactant a cikin kwalbar Erlenmeyer mai milimita 500 da ke ɗauke da 200 mL na maganin Bushnell Haas mai tsafta wanda aka ƙara masa man fetur 1% a matsayin tushen carbon. An yi allurar rigakafin Enterobacter cloacae SJ2 (1.4 × 104 CFU/ml) kafin a fara noma shi a kan injin girgiza a 37°C, 200 rpm na tsawon kwanaki 7. Bayan lokacin shiryawa, an cire biosurfactant ta hanyar amfani da centrifuging na maganin a 3400 ×g na tsawon mintuna 20 a 4°C kuma an yi amfani da supernatant da aka samu don dalilai na tantancewa. An ɗauki hanyoyin ingantawa da kuma siffanta biosurfactants daga bincikenmu na baya26.
An samo tsutsotsin Culex quinquefasciatus daga Cibiyar Nazarin Ci Gaba a Kimiyyar Ruwa (CAS), Palanchipetai, Tamil Nadu (Indiya). An kiwon tsutsotsin a cikin kwantena na filastik da aka cika da ruwan da aka cire daga ion a zafin jiki na 27 ± 2°C da kuma lokacin daukar hoto na 12:12 (haske: duhu). An ciyar da tsutsotsin sauro da maganin glucose na 10%.
An gano tsutsotsin Culex quinquefasciatus a cikin tankunan feshi masu buɗewa da marasa kariya. Yi amfani da jagororin rarrabuwa na yau da kullun don gano tsutsotsin a cikin dakin gwaje-gwaje85. An gudanar da gwaje-gwajen kashe tsutsotsin bisa ga shawarwarin Hukumar Lafiya ta Duniya 86. SH. An tattara tsutsotsin quinquefasciatus na huɗu a cikin bututun da aka rufe a cikin ƙungiyoyi na 25 ml da 50 ml tare da tazara ta iska ta kashi biyu bisa uku na ƙarfinsu. An ƙara Biosurfactant (0-50 mg/ml) a kowane bututu daban-daban kuma an adana shi a zafin jiki na 25 °C. Bututun sarrafawa ya yi amfani da ruwa mai narkewa kawai (50 ml). An ɗauki tsutsotsin da suka mutu a matsayin waɗanda ba su nuna alamun yin iyo ba a lokacin lokacin shiryawa (awanni 12-48) 87. Lissafa kashi na mace-macen tsutsotsin ta amfani da lissafin. (1)88.
Iyalin Odontotermitidae sun haɗa da termite na Indiya Odontotermes obesus, wanda aka samo a cikin katako masu ruɓewa a Harabar Noma (Jami'ar Annamalai, Indiya). Gwada wannan biosurfactant (0–50 mg/ml) ta amfani da hanyoyin da aka saba don tantance ko yana da illa. Bayan bushewa a cikin iska mai laminar na tsawon mintuna 30, an shafa kowane tsiri na takarda Whatman da biosurfactant a cikin taro na 30, 40, ko 50 mg/ml. An gwada tsiri na takarda da aka riga aka shafa da wanda ba a shafa ba kuma aka kwatanta su a tsakiyar farantin Petri. Kowane farantin petri ya ƙunshi kusan tururuwa masu aiki guda talatin O. obesus. An ba da takardar sarrafawa da gwaji a matsayin tushen abinci. An ajiye dukkan faranti a zafin ɗaki a duk tsawon lokacin shiryawa. Tururuwa sun mutu bayan awanni 12, 24, 36 da 488. Sannan an yi amfani da Daidaito na 1 don kimanta kashi na mace-macen tururuwa a yawan biosurfactant daban-daban. (2).
An ajiye samfuran a kan kankara kuma an naɗe su a cikin ƙananan bututun da ke ɗauke da 100 ml na sodium phosphate buffer mai nauyin 0.1 M (pH 7.4) sannan aka aika su zuwa Cibiyar Nazarin Cututtukan Aquaculture ta Tsakiya (CAPL) ta Cibiyar Nazarin Aquaculture ta Rajiv Gandhi (RGCA). Dakin Gwaji na Histology, Sirkali, Mayiladuthurai. District, Tamil Nadu, Indiya don ƙarin bincike. Nan da nan aka gyara samfuran a cikin 4% paraformaldehyde a zafin jiki na 37°C na tsawon awanni 48.
Bayan an gama gyaran, an wanke kayan sau uku da sinadarin sodium phosphate buffer mai nauyin 0.1 M (pH 7.4), an cire ruwa daga jiki a cikin ethanol sannan aka jika shi a cikin resin LEICA na tsawon kwanaki 7. Sannan a sanya sinadarin a cikin wani roba da aka cika da resin da polymerizer, sannan a sanya shi a cikin tanda mai zafi zuwa 37°C har sai tubalin da ke dauke da sinadarin ya zama polymer gaba daya.
Bayan an yi polymerization, an yanke tubalan ta amfani da LEICA RM2235 microtome (Rankin Biomedical Corporation 10,399 Enterprise Dr. Davisburg, MI 48,350, Amurka) zuwa kauri na mm 3. An raba sassan a kan zamiya, tare da sassa shida a kowane zamiya. An busar da zamiya a zafin ɗaki, sannan aka yi mata fenti da hematoxylin na minti 7 sannan aka wanke da ruwan gudu na minti 4. Bugu da ƙari, a shafa maganin eosin a fata na minti 5 sannan a wanke da ruwan gudu na minti 5.
An yi hasashen guba mai tsanani ta amfani da halittun ruwa daga wurare daban-daban na wurare masu zafi: kifi LC50 na awanni 96, D. magna LC50 na awanni 48, da kuma algae kore na awanni 96 EC50. An tantance gubar da ke tattare da rhamnolipid biosurfactants ga kifi da algae kore ta amfani da software na ECOSAR sigar 2.2 don Windows wanda Hukumar Kare Muhalli ta Amurka ta haɓaka. (Akwai shi akan layi a https://www.epa.gov/tsca-screening-tools/ecological-struct-activity-relationships-ecosar-predictive-model).
An gudanar da duk gwaje-gwajen aikin kashe ƙwari da kuma hana ƙwari sau uku. An gudanar da bayanan mace-macen tsutsa da ƙwari don ƙididdige matsakaicin yawan mutuwa (LC50) tare da tazara ta amincewa da kashi 95%, kuma an samar da lanƙwasa na mayar da hankali ta amfani da Prism® (sigar 8.0, GraphPad Software) Inc., Amurka) 84, 91.
Wannan binciken ya bayyana yuwuwar ƙwayoyin cuta masu rai da Enterobacter cloacae SJ2 ke samarwa a matsayin magungunan kashe ƙwari da kuma maganin kashe ƙwari, kuma wannan aikin zai taimaka wajen fahimtar hanyoyin da ake bi wajen magance ƙwari da kuma maganin kashe ƙwari. Nazarin histological na tsutsotsi da aka yi wa magani da biosurfactants ya nuna lalacewar hanyoyin narkewar abinci, tsakiyar hanji, cerebral cortex da hyperplasia na ƙwayoyin epithelial na hanji. Sakamako: Kimanta guba na aikin antitermite da larvicidal na rhamnolipid biosurfactant da Enterobacter cloacae SJ2 ya samar ya nuna cewa wannan keɓewa yana iya zama maganin kashe ƙwari don magance cututtukan da sauro ke haifarwa (Cx quinquefasciatus) da termites (O. obesus). Akwai buƙatar fahimtar gubar muhalli na biosurfactants da tasirinsu na muhalli. Wannan binciken yana ba da tushen kimiyya don tantance haɗarin muhalli na biosurfactants.
Lokacin Saƙo: Afrilu-09-2024