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Les 03 04 2 Rokyt.Indd JOURNAL OF FOREST SCIENCE, 50, 2004 (3): 109–117 Feeding of the willow leaf beetle Lochmaea capreae L. (Coleoptera, Chrysomelidae) on leaves of birch (Betula pendula Roth) contaminated by heavy metals L. ROKYTOVÁ, E. KULA, L. KODAROVÁ, A. PEŠLOVÁ Faculty of Forestry and Wood Technology, Mendel University of Agriculture and Forestry, Brno, Czech Republic ABSTRACT: The effect of contamination of birch (Betula pendula Roth) leaves with heavy metals on the feeding of imagoes of the willow leaf beetle (Lochmaea capreae L.) was studied under laboratory conditions. The imagoes preferred feeding on leaves less contaminated by Cd, Mn and Zn. The Pb content was tolerated on all the studied levels. The repellent effect of Zn 8,000 µg/ml in the Pb + Zn regime was compensated by Pb 500 μg/ml; the effect of Cd 250 μg/ml in relation to Mn 10,000 μg/ml was similar. Keywords: Lochmaea capreae; cadmium; manganese; lead; zinc; feed preference; laboratory conditions The willow leaf beetle (Lochmaea capreae L., 1758, mechanisms of the body eliminating heavy metals is, Chrysomelidae) occurs in Europe, Lapland, the British in addition to secretion in the excrements and exuviae Isles, the Balkans and Lake Baikal (PFEFFER 1954). Ima- and deposition in the skin or body organs, a capacity goes winter in tufts of grass or in plant litter; in early May, to search for and consume less contaminated feed (e.g. when average daily temperatures rise above 12°C, they HELIÖVAARA, VÄISÄNEN 1990; LINDQVIST 1992; begin maturation feeding (BROVDIJ 1973). The imagoes WEISMANN, ŘEHÁKOVÁ 1993; HRDLIČKA et al. 1999). make holes (windows) in the leaves of the genera Salix If the content of heavy metals in feed increases, on oral and Betula that damage them, and they naturally prefer contact with the contaminated part the insects stop feed- leaves higher up in the crown. They also attack the buds ing, they do not consume the feed, or they choose the less and bark of annual shoots (URBAN 1981). After repeated contaminated or not contaminated parts (WEISMANN et copulation (VI) the female lays 85–190 eggs in the soil at al. 1983). The interruption of feeding, lower feed con- the foot of the tree in clusters of 10–25 eggs. Embryonic sumption, metabolic disorders and the use of the feed development takes 1–2 weeks. Negative phototropic lar- may indirectly intensify the impact of heavy metals on vae skeletonise the leaves from the bottom side (VII to the physiology (development, fertility, enzyme activity, VIII). After 3–4 weeks they pupate in the surface soil lay- energy metabolism, deformation, behaviour, growth, ers and 10 days later the imagoes of the new generation respiration, vitality, mortality), ecology (abundance, leave the pupal chamber and begin maturation feeding diversity, density, competition) and insect evolution (in- (VIII–1⁄2 IX). In the Czech RepublicL. capreae rears one dustrial melanisms, resistance) (STARÝ, KUBIZŇÁKOVÁ generation (URBAN 1981). In 1985–1987 it gradated in 1987; HELIÖVAARA, VÄISÄNEN 1993; HABUŠTOVÁ, young birch stands established in the air-polluted area of WEISMANN 2001). the Krušné hory Mts. and the Děčín Sandstone Uplands Imagoes of the willow leaf beetle (L. capreae) feeding (KULA 1988). on birch (B. pendula) were used in laboratory conditions In the terrestrial food chain the phytophagous insects to specify the degree of preference to untreated and are an important link in the distribution of heavy met- heavy-metal-contaminated feed under different levels of als (KOWALCZYK, WATALA 1989). Among the defence application. Supported by the Grant research projects MSM 434100005 and NAZV 1144 funded by Ministry of Agriculture and by the following firms and companies: Netex; Alcan extruded products; Municipal Offices all in Děčín, Setuza in Ústí nad Labem, CEZ Ltd, Lafarge Cement Works in Čížkovice, North-Bohemian Mines in Chomutov, Dieter Bussmann in Ústí nad Labem. J. FOR. SCI., 50, 2004 (3): 109–117 109 Table 1. Spatial arrangement of the sequence Regressive Progressive Parallel metal leafa (µg/ml) metal leafa (µg/ml) level leafa metal (μg/ml) 1 2 1 250 1 Pb 4 2 10 2 50 2 Mn 100 Cd Cd low 3 50 3 10 3 Cd 2 4 250 4 2 4 control 0 1 100 1 10,000 1 Pb 20 2 500 2 5,000 2 Mn 500 Mn Mn increased 3 5,000 3 500 3 Cd 10 4 10,000 4 100 4 control 0 1 4 1 500 1 Pb 100 2 20 2 100 2 Mn 5,000 Pb Pb high 3 100 3 20 3 Cd 50 4 500 4 4 4 control 0 1 0 1 0 1 Pb 500 2 0 2 0 very 2 Mn 10,000 Control control 3 0 3 0 high 3 Cd 250 4 0 4 0 4 Control 0 a vertical position of the leaf from the upper part of the twig (1, 2 – apical; 3, 4 – basal leaves) Table 2. Area arrangement of the sequence Individual Parallel Combined metal (μg/ml) level metal (μg/ml) metals (μg/ml) Cd 0 low control 0 Pb + Zn 0 2 Cd 2 4 + 80 10 Mn 100 500 + 8,000 50 Pb 4 4 + 8,000 250 Zn 80 500 + 80 Mn 0 increased control 0 Mn + Cd 0 100 Cd 10 100 + 2 500 Mn 500 10,000 + 250 5,000 Pb 20 100 + 250 10,000 Zn 400 10,000 + 2 Pb 0 high control 0 Cd + Zn 0 4 Cd 50 2 + 80 20 Mn 5,000 250 + 8,000 100 Pb 100 2 + 8,000 500 Zn 4,000 250 + 80 Zn 0 very control 0 80 high Cd 250 400 Mn 10,000 4,000 Pb 500 8,000 Zn 8,000 110 J. FOR. SCI., 50, 2004 (3): 109–117 J. FOR. SCI., 50, 2004 (3): 109–117 111 Table 3. Scale of qualified estimate of feeding Table 4. Average content of heavy metals in birch leaves (μg/g) Degree Feeding (%) Feed 0 without feeding 0 Metal contamination Cd Mn Pb Zn (μg/ml) 1 poor feeding < 10 Distilled 0 0.26 124.8 2.11 148 2 medium feeding 10–25 watera 3 heavy feeding 25–50 Controlb 0 0.26 147.2 2.33 203 4 very heavy feeding 50–75 2 56.5 153 5.87 5 defoliation > 75 10 120 115 1.65 Cd 50 213 175 1.39 MATERIAL AND METHODS 250 347.5 377 1.24 100 0.38 348 1.85 Imagoes of the willow leaf beetle (L. capreae) col- 500 0.51 1,072 7.10 lected in the field in early spring (Sněžník, altitude Mn 600 m) were reared in a laboratory (temperature 20°C, 5,000 0.31 9,823 1.83 relative humidity 60%, normal light regime). They 10,000 0.71 11,740 3.56 were fed leaves of birch (B. pendula) contaminated 4 0.65 90.9 91.7 by steeping in solutions containing heavy metals (Cd 20 0.81 74.9 351 2–250, Mn 100–10,000, Pb 4–500, Zn 80–8,000 μg/ml). Pb Agrovital (ANONYM 1992) was used as a soaking agent 100 1.21 77.2 731 to ensure better adhesion of the solution. The control 500 9.30 145 1,035.5 leaves were steeped in Agrovital-containing distilled 80 0.38 93.7 3.68 340 water. After drying up the treated leaves were placed 400 0.23 89.6 5.98 967 into breeding vessels and changed in 24-hour intervals. Zn Each variant had three replications. Dead specimens 4,000 0.36 98.4 3.03 5,894 were continuously replaced by specimens from main- 8,000 0.39 92.4 4.35 13,066 tenance breeding. 4 + 80 1.45 132 28.7 271 The imagoes were removed into 4 l jars in a 20 ♀♀:6 ♂♂ 500 + 8,000 0.89 112 2,795 5,949 ratio and they were repeatedly given a twig with 4 con- Pb + Zn taminated leaves (spatial arrangement). With a regressive 4 + 8,000 5.41 104 23.9 6,751 sequence of the heavy metal on the leaves the concentra- 500 + 80 1.09 102 1,515 529 tion decreased from the lowest position to the top of the twig. With a progressive sequence the arrangement was 100 + 2 9.89 201 1.89 164 opposite. Twigs with uncontaminated leaves in two rep- 10,000 + 250 474 10,143 5.73 175 Mn + Cd lications were used as controls. A set of 4 leaves of the 100 + 250 616 262 2.96 187 twig made up a parallel sequence to the control below; 10,000 + 2 5.66 9,695 5.27 157 Cd, Mn and Pb-treated leaves were towards the top of the twig. The respective replications differed in the level of 2 + 80 10.3 108 1.56 355 feed contamination (low, increased, high and very high) 250 + 8,000 528 120 2.3 5,982 Cd + Zn (Table 1). 2 + 8,000 10 88.3 2.52 6,721 In order to eliminate the apical phenomenon, we chose 250 + 80 618 110 2.28 299 the area arrangement of leaves in Petri dishes (ø 16.5 cm) with filter paper on the bottom, and the willow leaf beetle a natural leaf composition, bdistilled water + Agrovital in a 15 ♀♀:5 ♂♂ ratio. Leaves were contaminated in an individual sequence of the heavy metal, in a set of control leaf and leaves treated with parallelly increased concentra- RESULTS tion, with a combination of two metals (Pb + Zn, Mn + Cd, Cd + Zn) respectively (Table 2).
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