Is nano ZnO/chlorpyrifos mixture more harmful to earthworms than bulk ZnO? A multigeneration approach.

As chlorpyrifos is one of the most widely used organophosphorus insecticides and ZnO-NPs are identified as NPs of the highest concern due to their negative effects on aquatic and soil organisms the objective of this study was to evaluate mixture toxicity of CHP and ZnO (bulk and nanoparticles (20 nm)) on two types of soil, artificial (AS) and natural (NS), and over two generations of earthworms. Primary endpoint measured was reproduction inhibition and biochemical biomarkers (acetylcholinesterase, catalase, glutathione-S transferase and malondialdehyde content). Results showed that mixture toxicity differs in respects to all tested factors: soil type, ZnO particle size and earthworm generation. CHP/ZnO mixtures had synergistic effects and significantly reduced a number of juveniles in both generations in AS, while the effects were additive or even antagonistic in NS. There was no difference in reproduction inhibition in respect to particle size of ZnO used in the mixtures. Negative effects could also be detected on growth dynamics of juvenile earthworms (2nd generation) as they had lower initial body mas, reduced growth rate and lower body mass as adults. Measured enzymes responded differently in respect to ZnO particle size used in the mixtures, with CHP/bZnO producing stronger effects. Measured concentrations of the bioavailable Zn in the soils showed no difference in the concentration of bioavailable Zn2+ between mixtures, but significantly more Zn2+ was retrieved from AS. General biomarker response indicated that 2nd generation of earthworms had lower capability to cope with oxidative stress.

Effects of chronic dietary exposure to a low-dose of Malathion, Aroclor-1254 and 3-methylcholanthrene on three biomarkers in male mice.

The aim of this research was to examine the applicability of some chronic toxicological tests in the determination of exposure to xenobiotics present in concentrations below No Observed Adverse Effect Level (NOAEL) and below the detection limit of analytical instruments. In the present experiment tested chemicals (Malathion, Aroclor-1254 and 3-methylcholanthrene (3-MC)) were mixed with wheat grains and given to male mice as feed over a period of 12 months. 7-ethoxyresorufin-O-deethylase (EROD) activity with the 3-MC and Aroclor-1254 treatments reached the peak at 9th month of exposure (26.7 and 42.4 pmol⁻¹ mg(prot)-⁻¹, respectively), while malathion did not have significant influence. Glutathione (GSH) level depletion was highest after three months of exposure. Unexpectedly, acetylcholinesterase (AChE) activity increased after treatment with malathion, an organophosphorous insecticide. In conclusion, low-level concentrations chronically administered exert certain effects on the levels of selected enzymes, e.g. biomarkers.

Influence of molting on efficacy of two functionally different larvicides: Bti and temephos.

The sensitivity of the second, third (L3), and fourth (L4) instars of Culex pipiens L. and Ochlerotatus sticticus (Meigen) to two larvicides, temephos and Bacillus thuringiensis variety israelensis (Bti) was tested with and without molting occurring in the experiments. In the first experiment for both tested mosquito species, LC50 values increased from the second to the fourth instar when tested against temephos, whereas for Bti the opposite trend was observed. The highest LC50 value was found for fourth instars of Cx. pipens (0.00405 mg/liter) against temephos and for second instars of Oc. sticticus (0.267 mg/liter) against Bti. The determined LC50 values for the second and third instars of both species decreased with an increased number of molted larvae in the experiments with temephos. For the experiments with Bti molting did not have any significant influence on LC50 values, except a small increase in toxicity during the L3/L4 molt of Oc. sticticus. These findings could help assess and define larviciding, as well as influence the quantity of larvicides needed for an efficient treatment.

Influence of honey bee products on transplantable murine tumours.

The effect of propolis [it is a water-soluble derivative (WSDP)] and related polyphenolic compounds of propolis (caffeic acid, caffeic acid phenethyl ester and quercetin), honey, royal jelly and bee venom on tumour growth, metastasizing ability and induction of apoptosis and necrosis in murine tumour models (mammary carcinoma and colon carcinoma) was investigated. WSDP and related polyphenolic compounds showed significant anti-metastatic effect (P < 0.01 and P < 0.001) given either before or after tumour-cell inoculation. Oral or systemic application of WSDP or caffeic acid significantly reduced subcutaneous tumour growth and prolonged the survival of mice. Honey also exerted pronounced anti-metastatic effect (P < 0.05) when applied before tumour-cell inoculation (peroral 2 g kg(-1) for mice or 1 g kg(-1) for rats, once a day for 10 consecutive days). Royal jelly did not affect metastasis formation when given intraperitoneally or subcutaneously. However, intravenous administration of royal jelly before tumour-cell inoculation significantly (P < 0.05) inhibited metastasis formation. When mice were given 10(5) tumour cells intravenously immediately after bee venom injection, the number of tumour nodules in the lung was significantly lower (P < 0.001) than in untreated mice or mice treated with bee venom subcutaneously. Local presence of bee venom in the tissue caused significant delay in subcutaneous tumour formation. These findings clearly demonstrate that anti-tumour and anti-metastatic effects of bee venom are highly dependent on the route of injection and on close contact between components of the bee venom and tumour cells. These data show that honey bee products given orally or systemically may have an important role in the control of tumour growth and tumour metastasizing ability.

Evaluation of the genotoxic and cytochrome P450 monooxygenase-inhibitory potential of Dicuran on procaryotic and eucaryotic test systems.

The effect of the herbicide Dicuran 500 FL (formulated product) on the phenotypical and genotypical changes in procaryotic and eucaryotic organisms was investigated using short-term tests for detecting genotoxins. Since pesticides discharged in the water environment can modulate the mixed-function monooxygenases (MFO) detoxification system of water organisms, the in vivo and in vitro effects of Dicuran on hepatic cytochrome P450 (cyt P450) monooxygenase activities were also examined in juvenile carp (Cyprinus carpio L.). By measuring the activities of MFO in experimental carp exposed to Dicuran an attempt was made to establish whether Dicuran could be bioactivated by MFO into ultimate mutagens. Our results on the bacterial strains Salmonella typhimurium TA100 and TA98 show that Dicuran does not possess either mutagenic or premutagenic characteristics. The micronucleus test on the erythrocytes of experimental carp did not establish any clastogenic effect either. However, Dicuran significantly inhibited the MFO activity of 7-ethoxyresorufin-O-deethylase (EROD) and benzo[a]pyrene monooxygenase (BaPMO) in the liver of experimental carp in vitro, as well as in in vivo conditions. These findings demonstrate the potentially damaging effect of Dicuran on the xenobiotic metabolizing enzyme systems of fish, and suggest the applicability of described methods for the prediction of the ecotoxicological significance of the presence of pesticides in the water environment.