Guthion insektisitinin Microtus canicaudus (Rodentia)'da etkisinin biyobelirteçler kullanılarak araştırılması
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Tarih
1993
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İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bir organofosforlu insektisit olan Guthion'un (azinphosmethyl) laboratuvar koşullarında barındırılan Microtus canicaudus (Kuzey Batı Amerika tarla faresi) 'da bazı biyobelirteçler kullanılarak, etkileri araştırıldı. Bu amaçla hayvanlardan alınan plazma örneklerinde laktat dehidrogenaz (LDH), kreatin kinaz (CK) ve izositrat dehid- rogenaz (ICDH) enzim aktiviteleri incelendi. Ayrıca plazma kreatinin ve kan üre nitrojeni (BUN) konsantrasyonları be lirlendi. Bundan başka asetilkolinesteraz enzim aktivitesi beyin doku homo jenatlarında incelendi. Kan akyuvar hücre farklılaşması oranları ve hematokrit değerleri insektisit etkisine bağlı olarak ele alındı. Vücut ağırlığı, besin ve su tüketimi gibi bazı toksikolojik parametreler hayvanla rın sağlık durumlarının belirlenmesinde değerlendirme kap samına alındı. Araştırmada tüm spektrofotometrik analiz yöntemleri için bir mikroplate okuyucu sistemi seçildi ve yöntemler bu cihaz için modifiye edildi. Hayvanların elimizdeki kimyasal ile muamelesi için iki ana test grubu oluşturuldu. Bunlardan birisi "Letal Konsantrasyon (LC)" test grubu, diğeri de "Disseksiyon (DS)" test grubu olarak adlandırıldı. Uygulama grupları olarak üç farklı insektisit konsantrasyonu seçildi ve bun lardan birisi M. canicaudus için LC50 dozu olarak bilinen 322 ppm'lik konsantrasyondu, insektisit dozlarından birisi daha düşük konsantrasyonda (161 ppm) ve diğeri de LC50 do zundan yüksek konsantrasyonda (428 ppm) seçildi. Herbir test grubu için ayrıca bir kontrol grubu oluşturuldu ve bu hayvanlar yalnızca dilusyon maddeleri olan aseton ve mısırözü yağı ile muamele edilmiş yem ile beslendiler. Test gruplarının her birini oluşturan uygulama gruplarında tüm hayvanlar her kafeste bir Microtus olacak şekilde barındırıldı. "LC" test grubu hayvanları deney süresince iki gün de bir kan örneklerinin alınması için kullanıldı. Deneyin son gününde (14. gün) canlı kalan hayvanlar ise dissekte edildi. "DS" test grubu hayvanlar rastgele seçilerek, yalnızca disseksiyon günlerinde kan örneklerinin alınmasını takiben öldürüldü ve beyin dokuları alındı. Yapılan deneysel çalışma sonuçlarına göre; a) Guthion etkisine bağlı olarak, vücut alırlığının her iki test grubunda da önemli düzeyde azaldığı saptandı (P<0.05). b) İnsektisit uygulama grupları ile kontrol grupları arasında, hematokrit değerleri bakımından farklılık saptanmadı. Ancak akyuvar farklılaşma yüzdesi verilerine göre, bazı gruplarda önemli istatistiksel farkliliklar bulundu (P<0.05). c) Guthion etkisi ile, beyin AChE aktivitesinin gerek "disseksiyon" ve gerekse "Letal Konsantrasyon" test grubunda eşey karışık (erkek ve dişi birlikte) ve erkek hayvanlarda önemli düzeyde inhibe edildiği (P<0.001; P<0.01; P<0.05), buna karşın dişilerde aktivite değişiminin kont rol grubu dişilerinden farklı olmadığı belirlendi. d) "DS" test grubu hayvanlarının kan plazması örneklerinde bazı farklı günlerde ve uygulanan farklı dozlarda LDH, CK, ICDH aktivitelerinin yada BUN konsantrasyonunun önemli düzeyde değiştiği bulundu (P<0.05). Benzer bulgular "LC" test grubu hayvanlardan alınan örneklerin çalışılması ile de bulundu. Ancak bu grup hayvanlarda bazı farklı uygulama günlerinde kreatinin düzeyinin de değiştiği gözlendi. Bununla beraber, tüm kan plazma biyokimyasal biyo- belirteçlerinin deneye alınmamış, sağlıklı stok laboratuvar hayvanlarındaki normal sınırları içerisinde olduğu saptandı. Gruplar arası istatistiksel farklılıklar varyans analizi ve Fisher 'in PLSD testi kullanılarak ortaya konuldu. Bulgular biyokimyasal, fizyolojik ve toksikolojik bazı biyobelirteç verileri kullanılarak tartışıldı. anahtar kelimeler: Guthion, Microtus canicaudus, biyobelirteç, asetilkolinesteraz, plazma enzimleri
Some biomarker parameters were investigated to detect the effects of Guthion (Azinphosmethyl), which is an orga- nophosphate insecticide on gray tailed voles (Hicrotus canicaudus) in the laboratory conditions. Lactate dehydro genase (LDH), creatin kinase (CK) and isocitrate dehydro genase (ICDH) enzyme activities and creatinin and blood urea nitrogen (BUN) concentrations were investigated in the plasma samples. Also brain acetylcholinesterase (AChE) activity was studied with brain tissue homogenates. On the other hand, white blood cell differentiation ratios and hematocrit values were investigated as blood physiological biomarkers of insecticide exposure. Some toxicological parameters, such as body weight, food and water consump tions, were considered to detect the health conditions of the animals. All biochemical experiments were assayed by using a microplate reader system for spectrophotoraetric techniques and assay methods were modified for this system. Two test groups were designed for chemical exposure. They called the "lethal concentration (LC) test" and the "sacrifice (DS) test" groups. Treatment groups were selected as three exposure concentrations, one of which was the LC50 (322 ppm) for voles. The other dose was lower (161 ppm) and the last dose was higher (428 ppm). Control group (diluent only) was also included each test groups. Animals were individually housed for the duration of the experiment for each treatment group of the test groups. V Lethal concentration test animals were bled every second day [on (-l)st, 2nd, 4th, 6th, 8th, 10th and 14th days] during the laboratory exposure using orbital bleeding techniques. If the animals are alive on day 14th, they were sacrificed for collection of brain tissues. But, sacrifice test animals were used for bleeding on the sacrifice day only (on day 3th, 7th or 9th) and those were selected randomly for this procedure. Finally, they were sacrificed to collect brain tissues. Analysis of variance (ANOVA) tests and Fisher's PLSD test were used to determine the significant differences among test groups compared to results of control groups. This experimental study results are shown that; a) Average body weight of both test group voles were significantly decreased because of Guthion exposure (P<0.05). b) Hematocrit values were not found significantly different between control and treatment groups. However, white cell differentiation ratios were shown significant differences for some treatment groups on the different test days. c) Brain AChE activity of both "sacrifice" and "LC" test group animals were significantly inhibited because of insecticide exposure for both sexes combined and male voles (P<0.001; P<0.01; P<0.05) but, enzyme activity were not significantly changed for females. d) Plasma LDH, CK, ICDH activities or BUN concentra tions of "sacrifice" test group animals were changed for different exposure doses on the different test days (P<0.05). Similar results were also detected for "LC" test group voles. But, creatinin concentrations of this test group animals were changed for different treatment days. However, biochemical results of blood plasma biomarkers were found in normal limits on the animals when they com pared to healty stock laboratory voles. Analysis of variance (ANOVA) and Fisher's PLSD tests were used for determine to test results. Experimental results were discussed using toxicological, biochemical and physiological biomarker criterias. keywords: Guthion, vole, biomarker, acetylcholinesterase plasma enzymes
Some biomarker parameters were investigated to detect the effects of Guthion (Azinphosmethyl), which is an orga- nophosphate insecticide on gray tailed voles (Hicrotus canicaudus) in the laboratory conditions. Lactate dehydro genase (LDH), creatin kinase (CK) and isocitrate dehydro genase (ICDH) enzyme activities and creatinin and blood urea nitrogen (BUN) concentrations were investigated in the plasma samples. Also brain acetylcholinesterase (AChE) activity was studied with brain tissue homogenates. On the other hand, white blood cell differentiation ratios and hematocrit values were investigated as blood physiological biomarkers of insecticide exposure. Some toxicological parameters, such as body weight, food and water consump tions, were considered to detect the health conditions of the animals. All biochemical experiments were assayed by using a microplate reader system for spectrophotoraetric techniques and assay methods were modified for this system. Two test groups were designed for chemical exposure. They called the "lethal concentration (LC) test" and the "sacrifice (DS) test" groups. Treatment groups were selected as three exposure concentrations, one of which was the LC50 (322 ppm) for voles. The other dose was lower (161 ppm) and the last dose was higher (428 ppm). Control group (diluent only) was also included each test groups. Animals were individually housed for the duration of the experiment for each treatment group of the test groups. V Lethal concentration test animals were bled every second day [on (-l)st, 2nd, 4th, 6th, 8th, 10th and 14th days] during the laboratory exposure using orbital bleeding techniques. If the animals are alive on day 14th, they were sacrificed for collection of brain tissues. But, sacrifice test animals were used for bleeding on the sacrifice day only (on day 3th, 7th or 9th) and those were selected randomly for this procedure. Finally, they were sacrificed to collect brain tissues. Analysis of variance (ANOVA) tests and Fisher's PLSD test were used to determine the significant differences among test groups compared to results of control groups. This experimental study results are shown that; a) Average body weight of both test group voles were significantly decreased because of Guthion exposure (P<0.05). b) Hematocrit values were not found significantly different between control and treatment groups. However, white cell differentiation ratios were shown significant differences for some treatment groups on the different test days. c) Brain AChE activity of both "sacrifice" and "LC" test group animals were significantly inhibited because of insecticide exposure for both sexes combined and male voles (P<0.001; P<0.01; P<0.05) but, enzyme activity were not significantly changed for females. d) Plasma LDH, CK, ICDH activities or BUN concentra tions of "sacrifice" test group animals were changed for different exposure doses on the different test days (P<0.05). Similar results were also detected for "LC" test group voles. But, creatinin concentrations of this test group animals were changed for different treatment days. However, biochemical results of blood plasma biomarkers were found in normal limits on the animals when they com pared to healty stock laboratory voles. Analysis of variance (ANOVA) and Fisher's PLSD tests were used for determine to test results. Experimental results were discussed using toxicological, biochemical and physiological biomarker criterias. keywords: Guthion, vole, biomarker, acetylcholinesterase plasma enzymes
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Anahtar Kelimeler
Biyoloji, Biology
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Scopus Q Değeri
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Künye
Özmen, M. (1993). Guthion insektisitinin Microtus canicaudus (Rodentia)'da etkisinin biyobelirteçler kullanılarak araştırılması. Yayımlanmış Doktora lisans tezi, İnönü Üniversitesi, Malatya.
