Interaction of Flupyradifurone and Deltamethrin, Two Pesticides Commonly Used for Plant Pest Control, in Honeybees.

Nowadays, old-generation pesticides are released into ecosystems alongside new formulations, giving rise to pharmacological interactions (additive, synergistic, and antagonistic effects). The aim of this study was to evaluate the impact that simultaneous exposure to DMT and FLU doses has on bee health. Groups of twenty honeybees were housed in cages to compose six macro-groups. One group consisted of experimental replicates treated orally with a toxic dose of deltamenthrin (DMT 21.6 mg/L); two other groups were subjected to the oral administration of two toxic doses of flupyradifurone (FLU 50 mg/L and FLU 100 mg/L); and two other groups were intoxicated with a combination of the two pesticides (DMT 21.6 + FLU 50 and DMT 21.6 + FLU 100). The consequences of the pesticides' interactions were highlighted by measuring and comparing data on survival, food consumption, and abnormal behavior. Generally speaking, antagonism between the two pesticides has been demonstrated. The bees were able to survive for up to three days at the lowest dosage of FLU (50 mg/L), with 46% of the subjects still alive; however, the maximum dose (100 mg/L) caused all treated animals to die as early as the second day. When DMT and FLU 50 were administered together, the group that received DMT alone had a lower survival rate. When comparing the survival rates produced by the DMT and FLU 50 combination to those of the group receiving FLU 50 alone, the same was clearly visible. While there was no statistically significant improvement observed when the survival indices of the DMT and FLU 100 combination were compared to those of the group intoxicated with DMT alone, an improvement in survival indices was observed when these were compared with the group intoxicated with FLU 100 alone.

Bergamot Polyphenolic Fraction for the Control of Flupyradifurone-Induced Poisoning in Honeybees.

Flupyradifurone (FLU) is a butenolide insecticide that has come onto the market relatively recently. It is used in agriculture to control aphids, psyllids, and whiteflies. Toxicity studies have decreed its low toxicity to honeybees. However, recent research has challenged these claims; oral exposure to the pesticide can lead to behavioral abnormalities and in the worst cases, lethal phenomena. Compounds with antioxidant activity, such as flavonoids and polyphenols, have been shown to protect against the toxic effects of pesticides. The aim of this research was to evaluate the possible protective effect of the bergamot polyphenolic fraction (BPF) against behavioral abnormalities and lethality induced by toxic doses of FLU orally administered to honeybees under laboratory conditions. Honeybees were assigned to experimental groups in which two toxic doses of FLU, 50 mg/L and 100 mg/L were administered. In other replicates, three doses (1, 2 and 5 mg/kg) of the bergamot polyphenolic fraction (BPF) were added to the above toxic doses. In the experimental groups intoxicated with FLU at the highest dose tested, all caged subjects (20 individuals) died within the second day of administration. The survival probability of the groups to which the BPF was added was compared to that of the groups to which only the toxic doses of FLU were administered. The mortality rate in the BPF groups was statistically lower (p < 0.05) than in the intoxicated groups; in addition, a lower percentage of individuals exhibited behavioral abnormalities. According to this research, the ingestion of the BPF attenuates the harmful effects of FLU. Further studies are needed before proposing BPF incorporation into the honeybees' diet, but there already seem to be beneficial effects associated with its intake.

Antimicrobial activity of essential oils (Citrus bergamia Risso & Poiteau, Melaleuca alternifolia and Chenopodium botrys) on pathogen strains isolated in milk samples from mastitic sheep.

The aim of this investigation was to study in vitro antibacterial activity of essential oils (EOs) (Citrus bergamia Risso & Poiteau, Melaleuca alternifolia and Chenopodium botrys) on pathogenic strains isolated in milk samples from mastitic sheep. The in vitro antibacterial activity showed that EOs induced higher bacterial inhibition against pathogens isolated in mastitic milk (Staphylococcus xylosus, Staphylococcus epidermidis, Streptococcus salivarius, Enterococcus faecium, Streptococcus agalactiae and Pseudomonas aeruginosa) than common antimicrobial agents used in veterinary medicine, with Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values significantly lower, excepted for P. Aeruginosa. The antibacterial activity of EOs has been attributed to the chemical composition of each phyto-complexes, constituted by monoterpenes, sesquiterpenes and their associated alcohols, identified by GC-MS analysis, with a synergic action in the bacterial cells. The results obtained confirmed in vitro antimicrobial activity of these EOs and their potential use in the treatment of sheep mastitis infection.

Protective Role of Bergamot Polyphenolic Fraction (BPF) against Deltamethrin Toxicity in Honeybees (Apis mellifera).

Pesticide-induced poisoning phenomena are a serious problem for beekeeping and can cause large losses of honeybee populations due to acute and sub-acute poisoning. The reduced responsiveness of honeybees to the damage caused by pesticides used in agriculture can be traced back to a general qualitative and quantitative impoverishment of the nectar resources of terrestrial ecosystems. Malnutrition is associated with a decline in the functionality of the immune system and the systems that are delegated to the detoxification of the organism. This research aimed to verify whether bergamot polyphenolic extract (BPF) could have protective effects against poisoning by the pyrethroid pesticide deltamethrin. The studies were conducted with caged honeybees under controlled conditions. Sub-lethal doses of pesticides and related treatments for BPF were administered. At a dose of 21.6 mg/L, deltamethrin caused mortality in all treated subjects (20 caged honeybees) after one day of administration. The groups where BPF (1 mg/kg) was added to the toxic solution recorded the survival of honeybees by up to three days. Comparing the honeybees of the groups in which the BPF-deltamethrin association was added to the normal diet (sugar solution) with those in which deltamethrin alone was added to the normal diet, the BPF group had a statistically significant reduction in the honeybee mortality rate (p ≤ 0.05) and a greater consumption of food. Therefore, it can be argued that the inclusion of BPF and its constituent antioxidants in the honeybee diet reduces toxicity and oxidative stress caused by oral intake of deltamethrin. Furthermore, it can be argued that BPF administration could compensate for metabolic energy deficits often induced by the effects of malnutrition caused by environmental degradation and standard beekeeping practices.

Toxicological analysis of metals content in agro-food wastes as possible supplement in animal feed.

The agro-food surplus is considered the most abundant renewable resource, interesting for the economic and environmental impact, with a possible employment in different productive fields such as supplement in animal feed. For a safe and effective enhancement of agro-food wastes it is needed a toxicological analysis before of their employment, to avoid potential risks to animals, humans and environment. The purpose of this study was to evaluate the metals content (Cu, Fe, Zn, Mg, Na, K, Ca, Mn, Se, Zn, Co, Ni, V, Sb, Mo, Cr and Pb, Cd, As, Ba), in agro-food wastes through ICP-MS analysis. The results obtained showed the presence of metals analysed with a significant content of essential micro and macro-elements; with regard to toxic metals, only strawberries and fennel samples showed Pb levels above the MRL. These data confirm, from the toxicological point of view, a possible and safe reuse of the most of all agro-food wastes, as raw material at low cost and rich of essential nutrients and particularly as supplement in animal feed.

Gourmet Table Salts: The Mineral Composition Showdown.

Table salts with their specialty flake size, textures, flavors, and colors can be considered a gastronomy niche food already increasing in demand worldwide. Being unrefined, they can contain trace elements potentially both healthy and toxic. In this study, 12 mineral elements (Al, Ca, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se, and Zn) in 10 different salts commercially available in southern Italy namely, Atlantic grey, Baule volante, Guerande, Hawaiian pink, Hawaiian black, Himalayan pink, Maldon, Mozia, Persian blue, and smoked salts were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) and thermal decomposition amalgamation-atomic absorption spectrophotometry (TDA-AAS). The concentration of mineral elements was variable according to the type of salt and its geographical origin. Co, Cr, Cu, Hg, Pb, and Se levels were tolerable and Al, Ca, Fe, Mn, Ni, and Zn ranged significantly among the samples. Persian Blue and Atlantic Grey salts showed elevated levels of Fe and Zn; their intake can be helpful in some specific conditions. Nevertheless, Ni levels were high in Persian Blue and Smoked salts. Pb exceeded the maximum level in all samples. Additional monitoring analyses of mineral contents in table salts are recommended for human health.

Organic contamination of Italian and Tunisian culinary herbs and spices.

Contaminants belonging to various classes, as polychlorobiphenyls, polycyclic aromatic hydrocarbons, organochlorine pesticides, organophosphorous pesticides, pyrethroid insecticides, fungicides, herbicides, synergists, carbamates, acaricides and insect growth regulators were simultaneously analysed by GC-MS/MS in 118 Italian and Tunisian culinary herbs and spices. The results obtained in Italian samples shown that laurel samples were the most contaminated with the presence of 15 residues on 140, whereas in rosemary (max value of 35 ng/g for cis-chlorfenyvinphos) and oregano (max value of 118.16 ng/g for ethion) some occasional residues can be observed, but always lower than the maximum residue levels; all the others samples shown no contamination. Among Tunisian samples, only rosemary contains a notably high content of pollutants exceeded the EU maximum residue limits (i.e., alachlor and phosalone with level of 359.2 and 43.3 ng/g, respectively), while oregano was determined to be free of contaminant residues. Considering the comparison among the different organic pollutants in Italian and Tunisian spices and herbs evaluated and the differences observed in this study a harmonization of regulation on contaminant residues in herbs and spices for human consumption should be needed, considering their increased use in diet and cooking.

Traceability of Opuntia ficus-indica L. Miller by ICP-MS multi-element profile and chemometric approach.

BACKGROUND: Opuntia ficus-indica L. Miller fruits, particularly 'Ficodindia dell'Etna' of Biancavilla (POD), 'Fico d'india tradizionale di Roccapalumba' with protected brand and samples from an experimental field in Pezzolo (Sicily) were analyzed by inductively coupled plasma mass spectrometry in order to determine the multi-element profile. A multivariate chemometric approach, specifically principal component analysis (PCA), was applied to individuate how mineral elements may represent a marker of geographic origin, which would be useful for traceability. RESULTS: PCA has allowed us to verify that the geographical origin of prickly pear fruits is significantly influenced by trace element content, and the results found in Biancavilla PDO samples were linked to the geological composition of this volcanic areas. It was observed that two principal components accounted for 72.03% of the total variance in the data and, in more detail, PC1 explains 45.51% and PC2 26.52%, respectively. CONCLUSION: This study demonstrated that PCA is an integrated tool for the traceability of food products and, at the same time, a useful method of authentication of typical local fruits such as prickly pear. © 2017 Society of Chemical Industry.

Monitoring of neonicotinoid pesticides in beekeeping.

The decline of pollinating species is correlated to the extensive use of neonicotinoids against pest insects for crop protection. In this study, the concentrations of neonicotinoid insecticides were determined in honeybees, honeycomb and honey samples, collected in Spring 2015 (blooming period) from different areas in Sicily (IT), to carry out an evaluation of bees products' safety and an overview of neonicotinoid contamination in beekeeping. The results obtained showed only the presence of clothianidin in bee samples and these concentrations don't represent a risk for bees' vitality and safety. The absence of residue in all honey samples, instead, showed the quality of bee products.

Mineral composition of some varieties of beans from Mediterranean and Tropical areas.

In this study has been evaluated the mineral composition (Cd, Pb, As, Hg, Ba, Cr, Co, Ni, Se, Sb, V, Cu, Fe, Zn, Mo, Na, K, Ca, Mg) of some varieties of bean (Phaseolus vulgaris, Vigna unguiculata and V. angularis) from Mediterranean and Tropical areas of the world (Italy, Mexico, India, Japan, Ghana and Ivory Coast); the correlation between beans mineral composition in micro and macroelements and botanical and/or geographical origin; trace elements dietary intake by beans consumption. The results showed a correlation between beans mineral composition and their geographical origin, with higher values in Ivory Coast samples. Moreover, minerals content found confirmed the importance of these legumes in the diet for the significant content of essential micro and macroelements and a safe consumption of beans for the low residual levels of toxic metals.

Are Biometric Parameters Helpful to Assess the Health Risk of Consuming Organochlorine Compounds Contaminated Silver European Eel (Anguilla anguilla)?

Several organochlorine compounds (OCs) were measured in European eels from the Tevere river (Italy). It followed that some of them are still important chemical contaminants. Concentrations of dichlorodiphenyltrichloroethanes (DDTs) are hazardous for the consumer health; those of the 6 indicator polychlorinated biphenyls (PCBs) are often close to the current European maximum residue limit and always higher than the former limit. The relationship between OC concentrations, biometric parameters and the lipid content was then investigated. A strong positive correlation with eel size emerged for the indicator PCBs and DDTs concentrations expressed on wet weight basis. This is explained by the corresponding higher lipid percentage that characterizes bigger eels and the absence of a dilution effect for compounds of main concerns. On the basis of the PCB-TDI threshold for a 70 kg person, we suggest that 1 should consume no more than 2 eels per week each weighing about 100 g. Thus, we conclude that eel consumption should be limited and restricted to eels relatively shorter and lighter.

Toxic Metals in Pelagic, Benthic and Demersal Fish Species from Mediterranean FAO Zone 37.

Fish represents a nutrient-rich food but, at the same time, is one of the most important contributor to the dietary intake of heavy metals. The aim of this study was to assess residual levels of Pb, Cd and Hg in different species, caught from FAO zones 37 1.3 and 37 2.2, particularly small pelagic, benthic and demersal fishes. The results obtained showed the absence of toxic metal in fishes from FAO zone 37 1.3. Relating to FAO zone 37 2.2, instead, in all samples we observed the absence of Pb, small concentrations of Cd (0.081±0.022 mg/kg) and higher Hg residual levels (0.252±0.033 mg/kg). Particularly, the trend of Cd contamination was similar in all species whereas Hg showed high levels in demersal, intermediate in pelagic and low in benthic species. However, only Cd concentrations exceed the MRL in mackerel, mullet, sea-bream fishes, according to Regulation CE n. 629/2008 and n. 488/2014.

Does the clam Ensis siliqua provide useful information about contamination by polychlorinated biphenyls and organochlorine pesticides beyond that of mussel Mytilus galloprovincialis?

Several polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) investigated in soft tissues of the frequently monitored Mytilus galloprovincialis were compared to those of Ensis siliqua, a highly dispersed and economically important bivalve species, though rarely investigated. Overall PCBs had higher concentrations than OCPs in both species with a prevalence of tri- tetra-and penta-chlorinated biphenyls in E. siliqua and a prevalence of hexa- hepta and octa-chlorinated biphenyls in M. galloprovincialis. E. siliqua emerges as a suitable complement to mussels for monitoring PCBs and OCPs pollution.

Feijoa sellowiana Berg fruit juice: anti-inflammatory effect and activity on superoxide anion generation.

Feijoa sellowiana Berg var. coolidge fruit juice was studied in vivo for the anti-inflammatory activity by carrageenin-induced paw edema test and in vitro for the effects on superoxide anion release from neutrophils in human whole blood. The fruit juice was analyzed by the high-performance liquid chromatography method, and quercetin, ellagic acid, catechin, rutin, eriodictyol, gallic acid, pyrocatechol, syringic acid, and eriocitrin were identified. The results showed a significant anti-inflammatory activity of F. sellowiana fruit juice, sustained also by an effective antioxidant activity observed in preliminary studies on 1,1-diphenyl-2-picrylhydrazyl (DPPH) test. In particular, the anti-inflammatory activity edema inhibition is significant since the first hour (44.11%) and persists until the fifth hour (44.12%) of the treatment. The effect on superoxide anion release was studied in human whole blood, in the presence of activators affecting neutrophils by different mechanisms. The juice showed an inhibiting response on neutrophils basal activity in all experimental conditions. In stimulated neutrophils, the higher inhibition of superoxide anion generation was observed at concentration of 10(-4) and 10(-2) mg/mL in whole blood stimulate with phorbol-myristate-13-acetate (PMA; 20% and 40%) and with N-formyl-methionyl-leucyl-phenylalanine (FMLP; 15% and 48%). The significant reduction of edema and the inhibition of O2(-) production, occurring mainly through interaction with protein-kinase C pathway, confirm the anti-inflammatory effect of F. sellowiana fruit juice.

Cytotoxic effects induced in vitro by organic extracts from urban air particulate matter in human leukocytes.

Urban areas represent major pollution sources as a result of anthropogenic activities located in these districts. Among the legislated air pollutants, polycyclic aromatic hydrocarbons (PAHs), which are mostly adsorbed on the surface of dust particles, are known for their adverse health effects. The present study has been carried out to examine the cytotoxic effects induced in vitro on human peripheral monocytes (PBMCs) by extractable organic matter (EOM) from PM10 (characterized for its PAH content) collected at four sites in the urban center of Messina, Italy. Chromatographic analyses showed the presence of PAHs in all EOM. Only EOM from one site induced a marked cell death probably resulting from the highest PAH content in this sample. Conversely, apoptosis activation was evident after PBMC exposure to all the EOM tested. These apoptotic effects do not appear related only to the total PAH content, but are probably influenced by chemical composition. In conclusion, our findings confirm that the cytotoxic potential of organic matter associated to ambient respirable air particles depends predominantly on the quantity and quality of the chemicals contained in it. In particular, the present data strongly evidence that the only evaluation of air concentration of particulate matter and benzo[a]pyrene, as well as the generally used risk models based on additivity, are not sufficient to evaluate air quality and PAH effect on human health because they do not take into account the possible inhibitory or synergic or antagonistic effect of combined exposure and the interference of other organic compounds present in respirable matter.

Possible use of Serratia marcescens in toxic metal biosorption (removal).

BACKGROUND, AIM AND SCOPE: Metal pollution is a serious problem for environmental safety and programmes of monitoring and bioremediation are needed. Among the processes of bioremediation, the use of microbes to remove and degrade contaminants is considered a biotechnological approach to clean up polluted environments. AIM: The aim of this study was to evaluate the ability of Serratia marcescens in Pb, Cd and Cr removal and the potential use of these bacteria in toxic metal bioremediation from polluted environments. METHODS: A short-term study (120 min) was carried out to study the bacterial growth in the presence of sub-inhibitory concentrations of each metal analysed and the kinetics of metal biosorption in S. marcescens strain. In addition, metal influence on the biosynthesis of the red pigment 'prodigiosina' by S. marcescens was monitored. RESULTS: The results obtained in this study show metals biosorption by S. marcescens (range: 0.0133-0.213 µg/g for Pb; 0.097-0.1853 µg/g for Cd; and 0.105-0.176 µg/g for Cr) and confirm the possible use of this bacterium to realize bioremediation processes, especially for Pb removal, and as a bioindicator of metal pollution.

Antioxidant properties of 4-methylcoumarins in in vitro cell-free systems.

4-methylcoumarins that possess two hydroxyl groups ortho to each other in the benzenoid ring have shown to have excellent antioxidant and radical-scavenging properties in different experimental models. Furthermore, they cannot be metabolized by the liver P450 monoxygenases and thus cannot form 3,4-coumarin epoxides, which are believed to be mutagenic. Herein, we present a study on the structure activity relationship of eight synthetic 4-methylcoumarins, carried out by employing a series of different chemical cell-free tests. These compounds were tested by means of three assays involving one redox reaction with the oxidant (DPPH assay, ABTS.+ assay and FRAP). Other assays were employed to evaluate the antioxidant properties of the coumarins under investigation against NO, O2.- and HClO, which are some of the major reactive oxygen and nitrogen species causing damage in the human body. Finally, we have measured the protective capacity of these coumarins against the oxidative damage in a simple biomimetic model of phospholipid membranes. Our results confirm the good antioxidant activity of the 7,8-hydroxy-4-methylcoumarins. In general, their activity is not significantly affected by the introduction of an ethoxycarbonylmethyl or an ethoxycarbonylethyl moiety at the C3 position. A discrete antioxidant activity is retained also by the 7,8-diacetoxy-4-methylcoumarins, although they are less efficient than the corresponding 7,8-dihydroxy compounds. Furthermore, as demonstrated in the brine shrimp toxicity test, none of the tested coumarins significantly affect the larvae viability. Two of the 4-methylcoumarins (7,8-dihydroxy-4-methylcoumarin and 7,8-dihydroxy-3-ethoxycarbonylethyl-4-methylcoumarin), very interestingly, showed strong scavenging activities against the superoxide anion and were also very effective in protecting the lipid bilayer against peroxidation. On the basis of these findings, these 4-methylcoumarins may be considered as potential therapeutic candidates for pathological conditions characterized by free radical overproduction.

Polychlorinated biphenyls and organochlorine pesticides in European eel (Anguilla anguilla) from the Garigliano River (Campania region, Italy).

Measuring organochlorine compounds in muscle tissue of European eels from the Garigliano River in Campania (Italy), overall PCBs emerged as the most abundant pollutants, followed by DDTs, Dieldrin and HCB. Target PCBs, IUPAC nos. 118, 138, 153 and 180, were the dominant congeners accounting for 64.2% of total PCBs. Among OCPs, p,p'-DDE was detected in all eels, always with higher concentration levels than other OCPs; p,p'-DDT was frequently detected, about 93.3% of the sample. The high and statistically significant correlations between concentration and length as well as weight of eels suggest that the organochlorine compounds concentrations tend to increase with the size and consequently with the age. Concentrations of DDTs and PCBs detected were similar to those reported in studies relative to France, UK and Sweden. As regards toxicological risk for human health, in general OCPs residual levels were below the limits established for fish and aquatic products. Conversely, the concentrations of PCBs exceeded the limit set by the EU for terrestrial foods. Our results imply that OCPs and PCBs are still important persistent chemical contaminants in Campania freshwaters, although their manufacture and use are banned or highly restricted.

Levels of benzo[a]pyrene and benzo[a]anthracene in smoked "Provola" cheese from Calabria (Italy).

Levels of benzo[a]pyrene (BaP) and benzo[a]anthracene (BaA) in "Provola" cheese samples from Calabria, smoked naturally and using commercial smoke-flavouring, have been determined. A comparative study of BaP and BaA concentrations was carried out on rind, exterior zone, core and slice of these samples. Quantitative determination of BaP and BaA was ascertained by HPLC using fluorescence detection. Levels of BaP and BaA were found in all smoked "Provola" cheese samples, but higher in cheese smoked with natural methods (0.46-1.13 µg kg⁻¹ for BaP and 1.38-9.29 µg kg⁻¹ for BaA) compared to samples smoked using commercial flavouring (0.085-0.32 µg kg⁻¹ for BaP and 1.20-2.98 µg kg⁻¹ for BaA). The smoking process contributes to polycyclic aromatic hydrocarbons contamination; however, for improved food safety, the use of commercial buffered smoke is preferable to traditional smoking procedures.

Influence of levetiracetam on the anticonvulsant efficacy of conventional antiepileptic drugs against audiogenic seizures in DBA/2 mice.

Levetiracetam (LEV, [S]-alpha-ethyl-2-oxo-1-pyrrolidine acetamide) is a new antiepileptic that has been used as adjunctive therapy to treat patients with intractable epilepsy. Systemic administration of levetiracetam (2.5-30 mg/kg, intraperitoneally (i.p.)) was able to produce a dose-dependent decrease in DBA/2 audiogenic seizure severity score. In combination with conventional antiepileptic drugs, levetiracetam, 5mg/kg, i.p., which per se did not significantly affect the occurrence of audiogenic seizures in DBA/2 mice, potentiated the anticonvulsant activity of some antiepileptic drugs studied against sound-induced seizures in DBA/2 mice. The degree of potentiation induced by levetiracetam was greater, approximately twice, for carbamazepine, diazepam, felbamate, topiramate, gabapentin, and valproate, less for lamotrigine, phenobarbital and phenytoin. This increase was associated with a comparable impairment in motor activity; however, the therapeutic index of combined treatment of antiepileptic drugs with levetiracetam was more favourable than the combination with saline with the exception of lamotrigine, phenytoin and phenobarbital. Since levetiracetam did not significantly influence the total and free plasma and the brain levels of antiepileptics studied. In addition, levetiracetam did not significantly affect the hypothermic effects of the anticonvulsants tested. In conclusion, levetiracetam showed an additive anticonvulsant effect when administered in combination with some classical anticonvulsants, most notably carbamazepine, diazepam, felbamate, gabapentin, topiramate and valproate, implicating a possible therapeutic relevance of such drug combinations.