A multibiomarker approach to evaluate the impact of anthropogenic contaminants on the ecotoxicological status of honey bees, Apis mellifera

A rapid decline of Apis mellifera, a keystone pollinator of wild plant species and agricultural crops, was recorded worldwide in recent years. The massive use of insecticides and fungicides in agriculture associated with pollution generated by other human activities and presence of parasites can cause toxicological effects in bees including a decrease of the immune defences, leading to the collapse of the colonies. Effective assessment of the ecotoxicological impacts of anthropogenic contaminants requires an approach that combines different biomarkers that enable a more precise diagnosis of exposure to environmental stressors through a combination of different biological responses. The aim of this study was to develop and apply a set of biomarkers to study the ecotoxicological status of honey bees. In the first phase, we investigated in the laboratory the effects of EMS, cadmium and a commercial fungicide (azoxistrobin 18.2% and ciproconazole 7.3%) in adult honey bees, evaluating eventual variation in glutathione S-transferase (GST), carboxylesterase (CaE), acetylcholinesterase (AChE) activities, alkaline phosphatase (ALP), lysozyme, erythrocyte nuclear abnormalities (ENA) assay and differential haemocytes count (DHC). Genotoxic effects, as well as alteration of the immune system, were found in bees treated with EMS, cadmium or the fungicide. Cadmium and the fungicide also inhibited AChE and CaE activities, GST was induced by all the compounds investigated. In the second phase, adult honey bees were collected from apiaries located in four environments characterized by different chemical input: a wooded environment (low input), an urban site, an orchard and a cultivated countryside site. Honey bees from the urban site were also collected and analyzed before and after treatment for parasites. ENA assay showed that bees taken from the countryside and the orchard had a greater number of abnormalities compared to the forest, confirming the presence of genotoxic substances in agricultural environments compared to control environments. GST activity was induced in bees from the urban environment, AChE was inhibited in the countryside compared to the forest, suggesting the presence of substances with neurotoxic effect in this environment. ALP activity was induced in all sites in comparison to wooded one. The bees collected after the parasites treatment showed an increase for GST activity as well as AChE inhibition

Multi-biomarker approach and IBR index to evaluate the effects of different contaminants on the ecotoxicological status of Apis mellifera

The honeybee, Apis mellifera L. (Hymenoptera: Apidae), a keystone pollinator of wild plant species and agricultural crops, is disappearing globally due to parasites and diseases, habitat loss, genetic constraints, beekeeper management issues and to the widespread use of pesticides. Besides insecticides, widely studied in this species, honeybees are also exposed to herbicides and fungicides and heavy metals whose lethal and sublethal effects need to be investigated. In this context, our study aimed to evaluate the effects of fungicides and of heavy metals on honeybees and to develop and apply a multi-biomarker approach that include an Integrated Biological Index (IBRv2) to assess the toxicological status of this species. Biomarkers of neurotoxicity (AChE and CaE), metabolic alteration (ALP, and GST) and immune system (LYS, granulocytes) were measured, following honeybees’ exposure to cadmium or to a crop fungicide, using the genotoxic compound EMS as positive control. A biomarker of genotoxicity (NA assay) was developed and applied for the first time in honeybees. At the doses tested, all the contaminants showed sublethal toxicity to the bees, highlighting in particular genotoxic effects. The data collected were analyzed by an IBRv2 index, which integrated the seven biomarkers used in this study. IBRv2 index increased with increasing cadmium or fungicide concentrations. The IBRv2 represents a simple tool for a general description of honeybees ecotoxicological health status. Results highlight the need for more in-depth investigations on the effects of fungicides on non-target organisms, such as honeybees, using sensitive methods for the determination of sublethal effects. This study contributes to the development of a multi-biomarker approach to be used for a more accurate ecotoxicological environmental monitoring of these animals

Multi-biomarker approach and IBR index to evaluate the effects of different contaminants on the ecotoxicological status of Apis mellifera

The honeybee, Apis mellifera L. (Hymenoptera: Apidae), a keystone pollinator of wild plant species and agricultural crops, is disappearing globally due to parasites and diseases, habitat loss, genetic constraints, beekeeper management issues and to the widespread use of pesticides. Besides insecticides, widely studied in this species, honeybees are also exposed to herbicides and fungicides and heavy metals whose lethal and sublethal effects need to be investigated. In this context, our study aimed to evaluate the effects of fungicides and of heavy metals on honeybees and to develop and apply a multi-biomarker approach that include an Integrated Biological Index (IBRv2) to assess the toxicological status of this species. Biomarkers of neurotoxicity (AChE and CaE), metabolic alteration (ALP, and GST) and immune system (LYS, granulocytes) were measured, following honeybees’ exposure to cadmium or to a crop fungicide, using the genotoxic compound EMS as positive control. A biomarker of genotoxicity (NA assay) was developed and applied for the first time in honeybees. At the doses tested, all the contaminants showed sublethal toxicity to the bees, highlighting in particular genotoxic effects. The data collected were analyzed by an IBRv2 index, which integrated the seven biomarkers used in this study. IBRv2 index increased with increasing cadmium or fungicide concentrations. The IBRv2 represents a simple tool for a general description of honeybees ecotoxicological health status. Results highlight the need for more in-depth investigations on the effects of fungicides on non-target organisms, such as honeybees, using sensitive methods for the determination of sublethal effects. This study contributes to the development of a multi-biomarker approach to be used for a more accurate ecotoxicological environmental monitoring of these animals

Can agricultural fungicides be a source of stress for non-target soil organisms? An ecotoxicological study on Eisenia fetida (Savigny, 1826)

Plant protection products, highly used in agriculture, can represent a source of stress for non-target soil organism and have a negative effect on ecosystems’ health. In particular, information on toxicological effects of fungicides are very poor or absent, despite their widespread use. The aim of this study was to evaluate the potential toxic effects of 4 commercial fungicides (Prosaro®, Amistar®xtra, Mirador® and Icarus®) on the earthworm Eisenia fetida (Savigny, 1826). Laboratory experiments were conducted using the filter paper test (FPT): E. fetida was exposed to increasing concentration of Prosaro® or Amistar®xtra, being the highest dose of treatment the recommended one for the usage in wheat farming. Field investigations were conducted transplanting E. fetida in cages in the soil of wheat and durum wheat fields before and during treatment with different combinations of the 4 fungicides. E. fetida specimens from laboratory and field work were analysed to evaluate vitality, potential neurotoxic effects (inhibition of acetylcholinesterase activity (AChE)), phase II enzymatic defense (glutathione S-transferase (GST)), oxidative stress (lipid peroxidation (LPO) and catalase (CAT) activity), genotoxic effects (Comet assay) and effect on the immune system (lysozyme activity). Laboratory studies with Prosaro® and Amistar®xtra showed alterations in organism’s vitality which increased with increasing treatment doses. Significant alteration of phase II metabolising enzymes (GST induction) and significant DNA fragmentation (Comet assay) with respect to controls were detected at environmentally relevant doses of Prosaro®. A statistically significant induction of GST was found in earthworms transplanted in the fields treated with Amistar®xtra alone and Amistar®xtra + Prosaro®. This study represents a first step towards a better understanding of commercial fungicides toxicological potential to non-target organisms. Data obtained indicate that deeper investigations are needed which should include long term artificial soil tests (AST) and further field studie

Risposte biochimiche nel granchio Pachygrapsus marmoratus per la valutazione della contaminazione ambientale del porto di Livorno (Italia).

The main purpose of the present investigation was to assess the health status of Livorno harbour, through the responses of a set of biomarkers in the crab Pachygrapsus marmoratus (Fabricius, 1787). This investigation is part of the IMPACT project (Port Impact on Marine Protected Area: crossborder co-operative actions), and the aim is to debug cross-borders management plans in order to actually protect the Marine Protected Areas. Male and female of the crabs were collected from two different areas: Livorno harbour, considered the polluted area, and the control area, the Marine Protected Area “Secche della Meloria”, located just a few miles from the Livorno harbour. A battery of biomarkers was employed to assess neurotoxic effects (acetylcholinesterase, AChE activity), energy metabolism (isocitrate dehydrogenease, IDH; lactate dehydrogenease, LDH), oxidative stress (lipid peroxidation, LPO; glutathione S-transferase, GST; glutathione peroxidase, GPX; glutathione reductase, GR; catalase, CAT; glutathione, GSH) and DNA damage (erythrocytic nuclear abnormalities, ENAs assay). Moreover, the levels of trace elements and PAHs were evaluated in the specimens. The results of LPO, CAT, GST, GSH and ENA assay showed that the crabs sampled in Livorno harbour are exposed to contaminants able to cause oxidative stress and genotoxic effects. The average values of LPO and ENA assay showed a statistically significant difference between specimens collected into Livorno harbour and those collected in the marine protected area. The average values of LPO were about three times higher in crabs sampled in polluted area in comparison to those sampled in the control area. Although the females showed higher values in comparison with the males, no significant differences between sex were found. A statistically significant difference was found between the two sampling sites regarding energetic metabolism biomarkers. The crab P. marmoratus, used as a bioindicator to investigate the contamination in a port area using a multi biomarker approach, can be considered a good model organism for marine coastal environment monitoring

First application of an Integrated Biological Response index to assess the ecotoxicological status of honeybees from rural and urban areas.

Understanding the effects of environmental contaminants on honeybees is essential to minimize their impacts on these important pollinating insects. The aim of this study was to assess the ecotoxicological status of honeybees in environments undergoing different anthropic pressure: a wood (reference site), an orchard, an agricultural area, and an urban site, using a multi-biomarker approach. To synthetically represent the ecotoxicological status of the honeybees, the responses of the single biomarkers were integrated by the Integrated Biological Response (IBRv2) index. Overall, the strongest alteration of the ecotoxicological status (IBRv2 = 7.52) was detected in the bees from the orchard due to the alteration of metabolic and genotoxicity biomarkers indicating the presence of pesticides, metals, and lipophilic compounds. Honeybees from the cultivated area (IBRv2 = 7.18) revealed an alteration especially in neurotoxicity, metabolic, and genotoxicity biomarkers probably related to the presence of pesticides, especially fungicides. Finally, in the urban area (IBRv2 = 6.60), the biomarker results (GST, lysozyme, and hemocytes) indicated immunosuppression in the honeybees and the effects of the presence of lipophilic compounds and metals in the environment

A multibiomarker approach to evaluate the impact of anthropogenic contaminants on the ecotoxicological status of honey bees, Apis mellifera

11noNumero articolo: 2.02.6reservedThe honeybee, Apis mellifera L. (Hymenoptera: Apidae), a keystone pollinator of wild plant species and agricultural crops, is disappearing globally due to parasites and diseases, habitat loss, genetic constraints, beekeeper management issues and to the widespread use of pesticides. Besides insecticides, widely studied in this species, honeybees are also exposed to herbicides and fungicides and heavy metals whose lethal and sublethal effects need to be investigated. In this context, our study aimed to evaluate the effects of fungicides and of heavy metals on honeybees and to develop and apply a multi-biomarker approach that include an Integrated Biological Index (IBRv2) to assess the toxicological status of this species. Biomarkers of neurotoxicity (AChE and CaE), metabolic alteration (ALP, and GST) and immune system (LYS, granulocytes) were measured, following honeybees’ exposure to cadmium or to a crop fungicide, using the genotoxic compound EMS as positive control. A biomarker of genotoxicity (NA assay) was developed and applied for the first time in honeybees. At the doses tested, all the contaminants showed sublethal toxicity to the bees, highlighting in particular genotoxic effects. The data collected were analyzed by an IBRv2 index, which integrated the seven biomarkers used in this study. IBRv2 index increased with increasing cadmium or fungicide concentrations. The IBRv2 represents a simple tool for a general description of honeybees ecotoxicological health status. Results highlight the need for more in-depth investigations on the effects of fungicides on non-target organisms, such as honeybees, using sensitive methods for the determination of sublethal effects. This study contributes to the development of a multi-biomarker approach to be used for a more accurate ecotoxicological environmental monitoring of these animals.mixedCaliani,I.; Campani, T.; Ammendola, A.; Conti, B.; Bedini, S.; Cosci, F.; Di Noi, A.; Gori, A.; Bellucci, F.; Giovanetti, L.; Casini, S.Caliani, I.; Campani, T.; Ammendola, A.; Conti, B.; Bedini, S.; Cosci, F.; Di Noi, A.; Gori, A.; Bellucci, F.; Giovanetti, L.; Casini, S