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Ovidius University Annals of Natural Sciences, Biology – Ecology Series Volume 14, 2010 CHANGES OF SOME PHYSIOLOGICAL PARAMETERS IN PRUSSIAN CARP UNDER THE ACTION OF SOME FUNGICIDE Maria Cristina PONEPAL, * Alina PĂUNESCU*, Alexandru Gabriel MARINESCU*, Octavian DRĂGHICI* * Universitatea din Piteşti, Facultatea de Ştiinţe Str. Tg. din Vale, nr.1 Piteşti, România, e-mail: ponepal_maria@yahoo.com __________________________________________________________________________________________ Abstract: This study was carried out to analyze the effects of sublethal and lethal concentrations of Bravo 500 SC, Champion 50 WP, Tilt 250 and Tiradin 70 PUS fungicide on some physiological parameters (oxygen consumption, breathing frequency, number of erythrocytes) of the prussian carp (Carassius auratus gibelio Bloch). The acute and subacute toxicity of fungicides was evaluated in glass aquaria under semi-static conditions. Keywords: prussian carp, fungicide, Bravo, Champion, Tilt, Tiradin, breathings frequency, oxygen consumption __________________________________________________________________________________________ 1. Introduction The commercial product Bravo 500 SC is a concentrated suspension of chlorothalonil (500g / l); chlorothalonil (2,4,5,6 tetrachlor isophthal-nitrile) is a contact fungicide with curative and preventive action (works by stopping germination and the development of spores) for combating a large number of pathogens (leaf spots, downy mildews, alternarioses, fruit rots, brown tor of fruit, scab) that threaten the main crops [1]. The fungicide is part of group IV of toxicity; it is not toxic to bees, warmblooded animals and moderately toxic to insects [2]. Chlorothalonil and its metabolites are very toxic to fish, aquatic invertebrates and marine organisms [3]: LC50 (96 h) is of 0.25 mg/l for rainbow trout (Salmo gairdneri), 0.3 mg/l for sun perch (Lepomis macrochirus), 0.43 mg/l for sea devil (Ictalurus punctatus), etc. Champion WP (copper hidroxide) is a fixed copper fungicide widely used for control of fungal and bacterial pathogens. Copper is highly toxic in aquatic environments and has effects in fish, invertebrates, and amphibians, with all three groups equally sensitive to chronic toxicity [4]. The Champion WP product is toxic to fish and aquatic organisms (96-hour LC50 Bluegill: 180 mg/l, 96-hour LC50 Rainbow trout: 0.023 mg/l and 48-hour EC50 Daphnia: 0.065 mg/l). Tilt 250 (the active ingredient is propiconazole – triazole fungicide) has protective, curative and systemic activity. Propiconazole's mode of action is demethylation of C-14 during ergosterol biosynthesis, and leading to accumulation of C-14 methyl sterols. The biosynthesis of these ergosterols is critical to the formation of cell walls of fungi [5]. The propiconazole is non toxic for bees, invertebrates and soil bacteriae, but is dangerous for fish and ather aquatic organisms (LC50 values ppm for freshwater fish species: bluegill 1.3-10.2, brown trout 3.5, rainbow trout 0.9-13.2, carp 6.8-21.0, catfish 2.0-5.1 and fathead minnow 7.6) [6] , [7]. Tiradin fungicide (the active substance is the thiuram - tetramethylthiuram disulphide TMTD) is a general use contact fungicide with protective action, third group of toxicity. Dithiocarbamates form a large group of chemicals that have numerous uses in agriculture and medicine [8]. It is used to control Botrytis on fruit and vegetables and in seed treatment. The 96-hour EC50 for algae growth inhibition is approximately 1 mg/l (1 ppm), the 48hour EC50 for Daphnia is less than 0.21 ppm and the 96-hour LC50 for fish is approximately 0.1 ppm (Bluegill sunfish, 0.0445 mg/l, Rainbow trout, 0.128 mg/l and 4 mg/l carp) [9]. This study was carried out to analyze the effects of sublethal and lethal concentrations – of some fungicide: Bravo 500 SC (from 0.078125 x 10 -3 to 12.5 x 10 -3 ml/l water), Champion 50 WP (from 0.003 to 3 mg/l water), Tilt 250 (from 0.25 to 4 ml/l water) and Tiradin 70 PUS (from 0.01 to 0.16 ml/l ISSN-1453-1267 © 2010 Ovidius University Press
Changes of some physiological parameters… / Ovidius University Annals, Biology-Ecology Series 14: 83-88 (2010) water) on some physiological parameters (oxygen consumption, breathing frequency, number of erythrocytes 0.078125 x 10 -3 and 1.5625 x 10 -3 ml Bravo/l water, 0.003 mg Champion/l water and 1 ml Tiradin/l water) of the prussian carp (Carassius auratus gibelio Bloch). The acute and subacute toxicity of this fungicide was evaluated in glass aquaria under semistatic conditions. 2. Material and Methods Determinations were made between January 2004 and October 2009 on prussian carp samples (Carassius auratus gibelio Bloch), captured from the surrounding rivers of Piteşti. Animals were acclimatized for 10 days before the completion of experiments in aquariums with a capacity of 100 l and 50 l [10], under conditions of natural photoperiodism, a period in which they were fed once a day (ad libitum), at around 10 am. After acclimatization in the laboratory, the fish were separated in two experimental variants (lots of 10-20 fish - average weight 18 g) subjected to fungicides. I. Determinations of oxygen consumption and frequency of respiratory movements at intervals of 24, 48, 72, 96, 168 and 336 hours on all samples of these lots (depending on survival) on prussian carp subjected to: - I.1. Bravo 500 SC in concentrations of 0.00078125, 0.0015625, 0.003125, 0.0625, 0.0125 ml /l water and the control lot - I.2. Champion 50 WP in concentrations of 0.003, 0.03, 0.3 and 3 mg/l water and the control lot Tilt 250 in concentrations of 0.25, 0.5, 1, 2, 4 ml /l water and the control lot - I.3. Tiradin 70 PUS in concentrations of 0.01, 0.02, 0.04, 0.08, 0.16 ml /l water and the control lot II. Hematological determinations (after one, respectively two weeks of exposure to the fungicide, the fishs were sacrificed to achieve intakes of blood necessary to hematological calculations (number of erythrocytes). 84 II.1 – fish subjected to Bravo 500 SC in concentrations of 0.00078125, 0.0015625, ml /l water and the control lot II.2 – fish subjected to Champion 50 WP in concentrations of 0.003 mg/l water and the control lot II.3 - fish subjected to Tilt 250 in concentration 1ml /l water and the control lot The fungicides concentrations were determined by preliminary tests of survival. The introduction of fish in solutions was done after their mixing and aeration for 5 minutes. The water temperature was 16-18°C, the "immersion" solution was changed every 24 hours, and aeration of water was continuous; the fish were not fed during experiments to avoid further intervention of this factor [10]. The testing method was systematic with refreshing solution at 24 hours after the calculations of the day, in aquariums of 100 l (50 l, respectively) for each experimental lot. Determination of oxygen consumption was done by means of the oximetre and Winkler method and erythrocytes were counted with Thoma chamber, using a small amount of blood from the caudal artery on the optic microscope [10], [11]. The statistical interpretation of the results was performed with ANOVA (LSD) test. 3. Results and Discussions The first four figures (fig.1-4) shows the average frequency of the respiratory movements of prussian carps exposed to the action of some fungicide (Bravo, Champion, Tilt and Tiradin).
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VOLUM OMAGIAL - Facultatea de Ştiinţe ale Naturii şi Ştiinţe Agricole

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