Malay. Appl. Biol. (December 2000) 29(1 &2): 61-67
EFFECT OF IN VITRO LEAD ON ACETYLCHOLINESTERASE ACTIVITY IN BRAIN TISSUE OF THE SNAKEHEAD FISH, HARUAN, CHANNA STRIATUS AND ABDUCTOR MUSCLES OF COCKLE, ANADARA GRANOSA
MAT-JAIS, A.M.,* HAMID, H.A., MOHAMED, M.Z. and ABDULLAH, A.S.
Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, MALAYSIA
ABSTRACT
This study was conducted to assess the effect of in vitro lead on acetylcholinesterase (AchE) activity in two selected indigenous tropical organisms, so as to establish the AchE as a biomarker. At the same time, the suitability of these rwo local aquatic species is also being evaluated as indicator for in situ environmental toxicology. A constant deterioration of local aquatic quality due to industrialization, urbanization, cross border dumping and other indiscriminate human activities, has prompted us to reevaluate the local environmental toxicological scenario. In this case reevaluation was carried out by assessing AchE changes in brain tissue of haruan and abductor muscle of cockles as biomarker. Haruan, Channa striatus, an important freshwater food fish, is commonly found near rice fields, small streams and ponds; while kerang, Anadara granosa, a popular seafood, is a marine invertebrate andsabundant along muddy coastal areas. Botb species are continuously and directly exposed to any contamination. In this experiment, we were using a cultured species of haruan weighing 72.20 + 12.55 gram and 20.65 ± 6.00 cm length and a random marketable sizes of cockles. Acethylcholinesterase activity in both samples were measured adopting a standard Ellman, et al., 1961 method using a Shimadzu UV 2401 PC spectrophotometer. Subsequently, the protein determination of the samples was carried out based on Lowry, et al., 1951 method. In this study, the level of AchE activity in C. striatus, was between 1.88 ± 0.027 pmolmnv'mg"1 protein control value and the lowest was - 0.220 + 0.160 pmolmiir'mg ' protein after exposure to 1 ppm in vitro lead for 30 min. Consequently, in cockles, A granosa, the level was between 4.277 ± 0.454 pmolmin 'mg-' protein control value and the lowest was - 0.901 ± 0.592 pmolmin-'gnr1 protein when exposed to a 30 min in vitro 1 ppm lead. Our result shows that lead inhibit AchE activity in both haruan brain tissue and cockles adductor muscles. The pattern of inhibition, was found to be dependent on dosage and duration of exposure, and the inhibition was significantly different at p < 0.01. Although, the inhibitory pattern in haruan, a freshwater fish is different from that of cockles, a marine organism; both tropical aquatic species appear to be a good candidates bioindicators AchE as the biomarker.
ABSTRAK
Penyelidikan ini telah dilakukan untuk menilai kesan in vitro plumbum ke atas aktiviti enzim asetilkolinesterase (AchE) _: dalam dua organisma asli tropika menggunakan AchE sebagai biomarker. Pada masa yang sama, kesesuaian kedua dua spesies ini juga dinilai sebagai indikator bagi toksikologi persekitaran in situ. Kemerosotan yang berterusan pada kualiti akuatik setempat yang disebabkan oleh perindustrian, perbandaran, pembuangan merentasi sempadan dan aktiviti - aktiviti tanpa dikriminasi manusia, telah mendorong kami untuk menilai semula senario toksikologi persekitaran setempat, dan di dalam kes ini, perubahan AchE di dalam tisu otak haruan dan otot abduktor kerang digunakan sebagai biomarker. Haruan, Channa striatus, adalah ikan airtawar dan makanan yang penting, dan biasanya ditemui di sawah padi, sungai kecil dan kolam; sementara kerang, Anadara granosa, merupakan makanan law yang popular, invertebrata laut dan banyak terdapat di sepanjang kawasan pantai berlumpur. Kedua-dua spesies ini sentiasa terdedah secara langsung dan berterusan kepada sebarang pencemaran. Di dalam eksperimen ini, spesies ikan haruan ternakan seberat 72.20 + 12.55g dan 20.65 ± 6.00 cm panjang digunakan, manakala kerang mengikut saiz rawak di pasar. Aktiviti AchE di dalam sampel telah diukur mengikut kaedah Ellman, et al., 1961 menggunakan spektrofotometer Shimadzu UV 2401 PC. Seterusnya, penentuan protein dalam sampel telah dilakukan megikut kaedah Lowry, et al., 1951. Di dalam penyelidikan ini, paras aktiviti AchE dalam C. striatus, adalah 1.88 + 0.027 pmolmin-'mg-' protein,. Nilai terendah adalah -0.220 ± 0.160 pmolmin-'mg-1 protein yang diperolehi selepas pendedahan selepas pendedahan kepada 1 ppm plumbum in vitro selama 30 min. Manakala, dalam A. granosa, parasnya adalah di antara 4.277 ± 0.454 pmolmin-mg-1 protein bagi nilai kawalan dan terendah adalah -0.901 + 0.592 pmolmin-'mg-1 protein selepas didedahkan kepada 1 ppm plumbum in vitro selama 30 min. Data ini menunjukkan bahawa plumbum telah merencat aktiviti AchE di dalam kedua - dua tisu otak haruan dan otot abduktor kerang. Perencatan yang diperolehi menunjukkan ia bergantung kepada kepekatan dan tempoh pendedahan, dan memberikan perbezaan bererti pada p<0.01. Corak perencatan dalam ikan airtawar, C.striatus adalah berbeza daripada organisma marin, A.granosa. Bagaimanapun, kedua spesies akuatik tropika ini adalah calon yang sesuai untuk dijadikan bioindikator dengan enzim AchE sebagai biomarker.
Key words: lead toxicity, inhibition, acetyl-cholinesterase activity, local indigenous species haruan, C. striatus, and cockle, Anadara granosa, test species.
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