- Email: [email protected]
Pathological and membrane biomarkers alteration induced by microcystins in liver, kidney and gills in tilapia fish (Oreochromis spp.)
Abstracts / Toxicology Letters 164S (2006) S1–S324
P16-08 Analysis of aflatoxin B1 in human hair samples O. Sabzevari 1 , I. Emrarian 1 , K.H. 3 Abouhossein , M. Amirahmadi 3
Abdi 2 , G.
1 Department
of Toxicology & Pharmacology, Tehran, Iran; 2 Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran University of Medical sciences, Tehran, Iran; 3 Food & Drug Control Labs, MOH, Tehran, Iran Aflatoxins, a group of related secondary metabolites of Aspergillus fungi, are commonly found on grains and seeds. Numerous outbreaks of acute aflatoxicosis have been documented worldwide and several epidemiological studies have shown a highly significant, positive correlation between human hepatic carcinoma and dietary aflatoxin contamination. Hair analysis has been introduced as an alternative or complementary method for testing the chemicals. The aim of this study was to identify aflatoxin B1 (AFB1), in human hair by HPLC. Fifty milligrams of hair samples, collected from healthy volunteers, was weighted and decontaminated by washing with 2 ml isopropanol, 2 ml distilled water (×3) and 2 ml isopropanol (1). The specimens were dried at room temperature and cut into small pieces. Thirty milligrams of the latter was incubated with AFB1 for 7 days at +4 ◦ C. After the incubation period, the samples were washed with isopropanol and distilled water (×2) until no AFB1 was present in washing liquid. The hair samples were then extracted using toluene/acetonitrile for 5 days at +4 ◦ C. The extraction phase was analysed by HPLC using fluorescence detector. By using the above procedure, we were able to detect AFB1 in hair samples which can be employed as tool to study patients’ history of exposure to aflatoxins. doi:10.1016/j.toxlet.2006.07.143 P16-09 Pathological and membrane biomarkers alteration induced by microcystins in liver, kidney and gills in tilapia fish (Oreochromis spp.) Silvia Pichardo 1 , Isabel M. Moreno 1 , Rafael Molina 1 , Angeles M. Jos 1 , Rosario Moyano 2 , Ana FerrerDufolt 3 , Ana M. Came´an 1 1 University of Seville Seville Spain; 2 Faculty of Veteri-
nary, University of Cordoba, Spain; 3 Toxicology Unit, Clinic University Hospital, Zaragoza, Spain Cyanobacterial blooms, which are considered a serious risk to animal and human populations, have been
S233
reported worldwide. Microcystins (MCs) are the most studied hepatotoxins produced by cyanobacteria. Toxic blooms of cyanobacteria, containing MCs, have been frequently associated with fish damage and mortality. In this study, membrane biomarkers and pathological changes produced by these toxins on hepatic, renal and gill tissues were investigated in Tilapia fish (Oreochromis spp.) under laboratory conditions. Four different groups of Tilapia were exposed to MC: the first group was injected intraperitoneally (i.p.) a dose of 900 g/kg of MCs (containing 500 g/kg MC-LR, 200 g/kg MCYR and 200 g/kg MC-RR), other two groups were injected i.p. doses of 500 g/kg MC-LR and 500 g/kg MC-RR, respectively, and a control group which was injected saline solution via i.p. Acid phosphatase (ACP), as a marker enzyme of lysosomal membranes, and alkaline phosphatase (ALP), as apical membrane enzyme, were analysed after exposure to MCs. Acute i.p. administration of all the exposures assessed produces a significant increase of alkaline phosphatase, whereas enhancement of acid phosphatase was only observed after exposure to MC-LR and to the mixture of MCs. The most severe histopathological changes were observed in kidney and liver, although gills were also affected. The most significant alterations observed were steatosis and necrosis in liver and necrotic epithelial cells and nephrosis in kidney. doi:10.1016/j.toxlet.2006.07.144 P17 Pesticide Toxicology P17-01 Optimization of DI-SPME combined with GC–MS for analyses of chlorophenoxypropionic acids R. Krzyzanowski, B. Leszczynski Department of Biochemistry, University of Podlasie, B. Prusa 12, Siedlce 08-110, Poland Solid phase microextraction (SPME) is one of the most promising method in analyzing herbicide residues. However, an application of SPME method requires determination of the proper analytical conditions. In the present paper, we report on optimization of the solid phase microextraction for analysis of such chlorophenoxypropionic herbicides like: 2-(4-chloro2-methylphenoxy)propionic acid (MCPP) and 2-(2,4dichlorophenoxy)propionic acid (2,4-DP). Seventy-five micrometer CAR/PDMS fiber was used to study an effect of time and temperature on the herbicide SPME (adsorption and desorption). The optimization was done for duration of adsorption (5–30 min) and
P16-08 Analysis of aflatoxin B1 in human hair samples O. Sabzevari 1 , I. Emrarian 1 , K.H. 3 Abouhossein , M. Amirahmadi 3
Abdi 2 , G.
1 Department
of Toxicology & Pharmacology, Tehran, Iran; 2 Department of Medicinal Chemistry, Faculty of Pharmacy, Tehran University of Medical sciences, Tehran, Iran; 3 Food & Drug Control Labs, MOH, Tehran, Iran Aflatoxins, a group of related secondary metabolites of Aspergillus fungi, are commonly found on grains and seeds. Numerous outbreaks of acute aflatoxicosis have been documented worldwide and several epidemiological studies have shown a highly significant, positive correlation between human hepatic carcinoma and dietary aflatoxin contamination. Hair analysis has been introduced as an alternative or complementary method for testing the chemicals. The aim of this study was to identify aflatoxin B1 (AFB1), in human hair by HPLC. Fifty milligrams of hair samples, collected from healthy volunteers, was weighted and decontaminated by washing with 2 ml isopropanol, 2 ml distilled water (×3) and 2 ml isopropanol (1). The specimens were dried at room temperature and cut into small pieces. Thirty milligrams of the latter was incubated with AFB1 for 7 days at +4 ◦ C. After the incubation period, the samples were washed with isopropanol and distilled water (×2) until no AFB1 was present in washing liquid. The hair samples were then extracted using toluene/acetonitrile for 5 days at +4 ◦ C. The extraction phase was analysed by HPLC using fluorescence detector. By using the above procedure, we were able to detect AFB1 in hair samples which can be employed as tool to study patients’ history of exposure to aflatoxins. doi:10.1016/j.toxlet.2006.07.143 P16-09 Pathological and membrane biomarkers alteration induced by microcystins in liver, kidney and gills in tilapia fish (Oreochromis spp.) Silvia Pichardo 1 , Isabel M. Moreno 1 , Rafael Molina 1 , Angeles M. Jos 1 , Rosario Moyano 2 , Ana FerrerDufolt 3 , Ana M. Came´an 1 1 University of Seville Seville Spain; 2 Faculty of Veteri-
nary, University of Cordoba, Spain; 3 Toxicology Unit, Clinic University Hospital, Zaragoza, Spain Cyanobacterial blooms, which are considered a serious risk to animal and human populations, have been
S233
reported worldwide. Microcystins (MCs) are the most studied hepatotoxins produced by cyanobacteria. Toxic blooms of cyanobacteria, containing MCs, have been frequently associated with fish damage and mortality. In this study, membrane biomarkers and pathological changes produced by these toxins on hepatic, renal and gill tissues were investigated in Tilapia fish (Oreochromis spp.) under laboratory conditions. Four different groups of Tilapia were exposed to MC: the first group was injected intraperitoneally (i.p.) a dose of 900 g/kg of MCs (containing 500 g/kg MC-LR, 200 g/kg MCYR and 200 g/kg MC-RR), other two groups were injected i.p. doses of 500 g/kg MC-LR and 500 g/kg MC-RR, respectively, and a control group which was injected saline solution via i.p. Acid phosphatase (ACP), as a marker enzyme of lysosomal membranes, and alkaline phosphatase (ALP), as apical membrane enzyme, were analysed after exposure to MCs. Acute i.p. administration of all the exposures assessed produces a significant increase of alkaline phosphatase, whereas enhancement of acid phosphatase was only observed after exposure to MC-LR and to the mixture of MCs. The most severe histopathological changes were observed in kidney and liver, although gills were also affected. The most significant alterations observed were steatosis and necrosis in liver and necrotic epithelial cells and nephrosis in kidney. doi:10.1016/j.toxlet.2006.07.144 P17 Pesticide Toxicology P17-01 Optimization of DI-SPME combined with GC–MS for analyses of chlorophenoxypropionic acids R. Krzyzanowski, B. Leszczynski Department of Biochemistry, University of Podlasie, B. Prusa 12, Siedlce 08-110, Poland Solid phase microextraction (SPME) is one of the most promising method in analyzing herbicide residues. However, an application of SPME method requires determination of the proper analytical conditions. In the present paper, we report on optimization of the solid phase microextraction for analysis of such chlorophenoxypropionic herbicides like: 2-(4-chloro2-methylphenoxy)propionic acid (MCPP) and 2-(2,4dichlorophenoxy)propionic acid (2,4-DP). Seventy-five micrometer CAR/PDMS fiber was used to study an effect of time and temperature on the herbicide SPME (adsorption and desorption). The optimization was done for duration of adsorption (5–30 min) and