Comparative evaluation of HET-CAM and ICE methods for objective assessment of ocular irritation caused by selected pesticide products.

Eye irritation potency of pesticides (fungicides, herbicides, insecticides) was comparatively tested by HET-CAM and ICE method. Based on the results of the tests the statistical analysis of agreement between classification using individual methods was done by Goodman-Kruskal's rank correlation and determination (calculation) of Cohen's kappa coefficient. Statistical analysis of agreement between classification revealed significant correlation between results of in vivo and in HET-CAM assays (76%). There was no significant correlation between result of in vivo and in ICE methods (64%). Weakest correlation was found between the data from in vitro HET-CAM and ICE tests. The percentage of agreement between two in vitro data was 48%. They may be recommended as a part of a battery of tests to reduce experimentation on mammals and to limit or eliminate pain and injury inflicted on experimental animals. The HET-CAM test is a useful tool for studying in vivo the potential conjunctival irritation, while the ICE test can be used to study corneal irritant effects in detail.

Monitoring of heavy metal burden in wild birds at eastern/north-eastern part of Hungary.

Concentrations of different heavy metals (As, Cd, Cr, Cu, Hg, Pb, Zn) were examined in the contour feathers of long-eared owl (Asio otus), little owl (Athene noctua), tawny owl (Strix aluco), barn owl (Tyto alba), Eurasian sparrowhawk (Accipiter nisus), rook (Corvus frugilegus), hooded crow (Corvus cornix), carrion crow (Corvus corone), common buzzard (Buteo buteo) and barn swallow (Hirundo rustica). The samples were collected from the Hortobágyi Madárpark (Bird Hospital Foundation) in Hungary. The bird species were classified into six groups based on their nourishment. Feathers were analysed by inductively coupled plasma optical emission spectroscopy (ICP-OES). The aim of our study was to determine the concentration of the above-mentioned heavy metals in the six different groups and to compare them by the groups, to find a possible connection between the concentrations and the age of birds and to get some information about the heavy metal burden of the environment. The highest As concentration was measured in little owl (0.65 ± 0.56 mg/kg). The highest Cd, Cr and Pb concentration was found in the feathers of barn swallow (0.13 ± 0.06 mg/kg; 1.69 ± 0.44 mg/kg; 5.36 ± 1.46 mg/kg), while the highest Cu and Hg concentration (65.45 ± 17.66 mg/kg; 2.72 ± 1.08 mg/kg) in sparrowhawk feathers and the highest Zn concentration in owls (157.21 ± 57.3 mg/kg). Statistically significant difference has been determined between the juvenile and adult crows in the case of Cd (p = 0.011). The higher concentration was measured in adults (0.14 ± 0.04 mg/kg) than that in juveniles (0.08 ± 0.02 mg/kg). Based on our results, the examined area is not contaminated by these heavy metals on that level, which can cause any adverse effect or poisoning in birds, so this region is safe to wildlife.

Monitoring of heavy metal burden in mute swan (Cygnus olor).

Concentrations of heavy metals (especially arsenic, cadmium, chromium, copper, mercury and lead) were measured in the contour (body) feathers of mute swans (Cygnus olor) and in its nutrients (fragile stonewort [Chara globularis], clasping leaf pondweed [Potamogeton perfoliatus], Eurasian watermilfoil [Myriophyllum spicatum], fennel pondweed [Potamogeton pectinatus]) to investigate the accumulation of metals during the food chain. The samples (17 feathers, 8 plants) were collected at Keszthely Bay of Lake Balaton, Hungary. Dry ashing procedure was used for preparing of sample and the heavy metal concentrations were analysed by inductively coupled plasma optical emission spectroscopy (ICP-OES). Copper (10.24 ± 2.25 mg/kg) and lead (1.11 ± 1.23 mg/kg) were detected the highest level in feathers, generally, the other metals were mostly under the detection limit (0.5 mg/kg). However, the concentrations of the arsenic (3.17 ± 1.87 mg/kg), cadmium (2.41 ± 0.66 mg/kg) and lead (2.42 ± 0.89 mg/kg) in the plants were low but the chromium (198.27 ± 102.21 mg/kg) was detected in high concentration.

Evaluation of mercury and lead content in the liver of the cormorant (Phalacrocorax carbo sinensis) population of Kis-Balaton, Hungary.

Mercury and lead concentrations were measured in the livers of cormorant (Phalacrocorax carbo sinensis), an aquatic bird species living and nesting in the special, highly protected nature conservation area of Kis-Balaton, Hungary. The measurements of metal concentrations were performed by atomic absorption spectrometry using the cold vapour method for mercury and the electrothermal method for lead. Mercury concentrations in the livers were significantly higher in the adult population (4.479 ± 3.336 mg/kg dry matter, DM) than in the juvenile birds (2.682 ± 2.087 mg/kg DM), indicating an increase of bioaccumulation with age. A similar pattern was not observed for lead. There were no statistical differences between males and females either in mercury or in lead concentrations. The average levels of mercury (3.580 ± 2.906 mg/kg DM) and lead (0.746 ± 0.499 mg/kg DM) were statistically different in the liver. No correlation was found between the concentrations of the two heavy metals. Recently, the wild birds have been chronically exposed to subtoxic amounts of metals which have a tendency to accumulate especially in the soft tissues.

HET-CAM test for determining the possible eye irritancy of pesticides.

Agrochemicals and veterinary products must undergo numerous toxicological tests before registration. The use of animals in these studies is a controversial issue. The Draize eye irritation test is one of the most criticised methods because of the injuries inflicted on the test animals. Several in vitro methods have been used to investigate the toxicity of potential eye irritants with a view to replacing in vivo eye irritation testing. One of these alternative methods is the Hen's Egg Test--Chorioallantoic Membrane (HET-CAM) test. In the present studies comparative screening was performed with a set of agrochemicals to establish parallel data on in vitro (HET-CAM) and in vivo (Draize) results. The examined materials were: Totril (ioxynil), Omite 57 E (propargit), Actellic 50 EC (pyrimiphos-methyl), Stomp 330 EC (pendimethalin), Mospilan 3 EC (acetamiprid), Alirox 80 EC (EPTC), Match 050 EC (lufenuron), Nonit (dioctyl sulphosuccinate sodium), Perenal (haloxyfop-R methyl ester), Pyrinex 48 EC (chlorpyrifos). These experiments showed good correlation between results obtained by the HET-CAM test and those of the Draize rabbit eye test in most cases. The present form of the HET-CAM test can be proposed as a pre-screening method for the determination of eye irritative potential, therefore the number of test animals can be reduced and/or experimental animals can be replaced.