In recent years, the awareness that pesticides can have other effects apart from generic toxicity is growing. In particular, several pieces of evidence highlight their influence on human fertility. In this study, we investigated, by a virtual screening approach, the binding between pesticides and proteins present in human gametes or associated with reproduction, in order to identify new interactions that could affect human fertility. To this aim, we prepared ligand (pesticides) and receptor (proteins) 3D structure datasets from online structural databases (such as PubChem and RCSB), and performed a virtual screening analysis using Autodock Vina. In the comparison of the predicted interactions, we found that famoxadone was predicted to bind Cellular Retinol Binding Protein-III in the retinol-binding site with a better minimum energy value of -10.4 Kcal/mol and an RMSD of 3.77 with respect to retinol (-7.1 Kcal/mol). In addition to a similar network of interactions, famoxadone binding is more stabilized by additional hydrophobic patches including L20, V29, A33, F57, L117, and L118 amino acid residues and hydrogen bonds with Y19 and K40. These results support a possible competitive effect of famoxadone on retinol binding with impacts on the ability of developing the cardiac tissue, in accordance with the literature data on zebrafish embryos. Moreover, famoxadone binds, with a minimum energy value between -8.3 and -8.0 Kcal/mol, to the IZUMO Sperm-Egg Fusion Protein, interacting with a network of polar and hydrophobic amino acid residues in the cavity between the 4HB and Ig-like domains. This binding is more stabilized by a predicted hydrogen bond with the N185 residue of the protein. A hindrance in this position can probably affect the conformational change for JUNO binding, avoiding the gamete membrane fusion to form the zygote. This work opens new interesting perspectives of study on the effects of pesticides on fertility, extending the knowledge to other typologies of interaction which can affect different steps of the reproductive process.

BACKGROUND: Vasovagal reaction (VVR) is a frequently encountered generalised donor adverse reaction, associated with donor deterrence towards future donation. Several mitigation strategies for prevention of VVR were tried but still not standardised. This quadri-armed randomised study evaluated the utility of water ingestion, applied muscle tension (AMT) and combination of both in preventing the VVR among blood donors.
METHODS: A quadri-armed randomised controlled trial was performed on 4320 whole blood donors. Blood donors of 18-65 years of age were randomised into four groups based on the interventions performed i.e., control with no intervention (Group 1, n = 1081), water ingestion (Group 2, n = 1082), AMT (Group 3, n = 1070) and combined intervention (Group 4, n = 1087). VVR during and immediately after blood donation were observed along with assessment of risk factors in blood donors and the effectiveness of interventions were analysed.
RESULTS: The incidence of VVR observed 1.6% in our study, with the highest occurrence in the control group (2.5%) and the lowest in the combined intervention group (0.9%). Multivariable logistic regression revealed that the control group donors faced a 1.38-fold greater risk of VVR compared to those receiving interventions (OR: 1.38, 95% CI: 1.10-1.75). Other risk factors included younger age (OR: 1.5, 95% CI: 1.05-2.17), first-time donation (OR: 5.7, 95% CI: 1.66-5.74), prior history of VVR (OR: 2.5, 95% CI: 10.4-101.52).
CONCLUSIONS: The combined approach of water ingestion and AMT proved significantly more effective in VVR prevention compared to individual interventions.

Strobilurins represent the most widely used class of fungicides nowadays andare considered relatively non-toxic to mammals and birds but highly toxic to aquatic biota. Dimoxystrobin is one of the novel strobilurins, recently included in the 3rd Watch List of the European Commission as available data indicate that it could pose a significant risk to aquatic species. As yet, the number of studies explicitly assessing the impact of this fungicide on terrestrial and aquatic species is extremely low, and the toxic effects of dimoxystrobin on fish have not been reported. Here we investigate for the first time the alterations induced by two environmentally relevant and very low concentrations of dimoxystrobin (6.56 and 13.13 μg/L) in the fish gills. morphological, morphometric, ultrastructural, and functional alterations have been evaluated using zebrafish as a model species. We demonstrated that even short-term exposure (96 h) to dimoxystrobin alters fish gills reducing the surface available for gas exchange and inducing severe alterations encompassing three reaction patterns: circulatory disturbance and both regressive and progressive changes. Furthermore, we revealed that this fungicide impairs the expression of key enzymes involved in osmotic and acid-base regulation (Na+/K+-ATPase and AQP3) and the defensive response against oxidative stress (SOD and CAT). The information presented here highlights the importance of combining data from different analytical methods for evaluating the toxic potential of currently used and new agrochemical compounds. Our results will also contribute to the discussion on the suitability of mandatory ecotoxicological tests on vertebrates before the introduction on the market of new compounds.

The vibrational study of the pyraclostrobin (methyl N-(2-{[1-(4-chlorophenyl)-1H-pyrazol-3-yl]oxymethyl}phenyl) N-methoxycarbamate) molecule, a synthetic fungicide for agricultural uses, was performed. Pyraclostrobin belongs to the strobilurin family and acts by inhibiting the fungus respiratory chain at the level of Complex III, becoming an excellent agent for preventive, curative and eradicative activities against a wide range of fungal plant pathogens. However, its presence needs to be monitored to avoid the excessive and/or improper use that may compromise human or environmental health. The FTIR and Raman spectra of pyraclostrobin in pure solid state were recorded and compared with those obtained from both the substance in CH2Cl2 solution and in an agricultural commercial product (Comet® BASF). The spectral analysis was complemented with quantum-chemical calculations at the DFT level (B3LYP/6-311G*) for the predictions of the molecular geometry and its vibrational behavior. The high flexibility of the molecule was explored by performing potential energy scans on several dihedral angles and the results suggested that the main conformer of pyraclostrobin is that possessing the ortho-substituted benzene ring in perpendicular orientation regarding the plane that contains the ether group and the pyrazole ring, although the presence of a second preferred conformation in the experimental vibrational spectra was not ruled out. Among the many vibrational bands of pyraclostrobin that were well identified in the spectrum of the composite product for agricultural use, the one located at 936 cm-1 stood out. This signal was assigned to a vibration of the pyrazole ring and promised to be a good candidate as marker of the presence of the fungicide in complex matrixes.

The chiral pesticide famoxadone is mainly applied to control fungal diseases on fruiting vegetables. The fungicidal activity, ecotoxicological effects, and degradation behavior of famoxadone enantiomers are less well known. In this study, a systemic assessment of the stereoselectivity of famoxadone was performed in cucurbits and soil. Famoxadone enantiomers presented distinct inhibitory activities among different fungal species. The bioactivities of R-(-)-famoxadone were 2.7-178 times higher than S-(+)-famoxadone toward five phytopathogens. Based on the obtained LC50 values, famoxadone was super toxic to Eisenia foetida (E. foetida). Moreover, the acute toxicity of R-(-)-famoxadone presented 167 times greater to E. foetida than that of S-(+)-famoxadone, indicating that R-(-)-famoxadone showed higher bioactivity toward target organisms and non-target organisms than S-(+)-famoxadone. In addition, a simple high-performance liquid chromatography (HPLC) method was established to determine the stereoselective degradation of famoxadone in two species of cucurbits (cucumber and chieh-qua) and in field soil. The half-life values of famoxadone degradation were from 5.4 to 14.1 days, indicating that famoxadone was easily degraded. Additionally, no stereoselective degradation was found in cucurbits and soil. The results may provide promising implications for comprehensive environmental and ecological risk assessments of famoxadone.

Despite the high usage of pesticides in current agricultural practices, its effects to humans and to the environment (non-target species) are a continuous concern. Soil dwelling organisms are among the first in line of exposure to pesticides, however their risks are often based on the pure active ingredient (a.i.) and not on the commercial formulated products (FPs) actually applied in the fields. In the present study, we investigated the effects of two fungicide FPs versus its a.i. (s): Amistar® XTRA and the respective a.i. (s) azoxystrobin and cyproconazole, and Prosaro® 250 EC and the respective a.i. (s) prothioconazole and tebuconazole, to the non-target soil oligochaete Enchytraeus crypticus. The standard Enchytraeid Reproduction Test was used to assess effects on survival and reproduction. Results showed that Amistar was more toxic than Prosaro, particularly for reproduction (EC50 = 161 mg Amistar/kg soil, EC50 = 350 mg Prosaro/kg soil). For both FPs, reproductive effects were mainly related to one of its a.i. (s) (azoxystrobin [EC50 = 37 mg azosxystrobin/kg soil] for Amistar, and tebuconazole [EC50 = 41 mg tebuconazole/kg soil] for Prosaro), while lethal effects were not predicted by the toxicity of its a.i. (s) (particularly in the case of Prosaro, which was more toxic than its a.i. (s)). These findings highlight the need to further explore the toxicity data of the FPs compared to the a.i. (s), aiming to predict a more realistic environmental hazard of pesticides.

Azoxystrobin, buprofezin, dinocap and hexaconazole are widely used in crop protection of mango from flowering to harvest. Residue assessment of these chemicals on mango fruits was done following treatments at the recommended and double doses as per good agricultural practices (GAP). Mango fruit and soil sample preparation was done by QuEChERS, and analysis was done using LC-MS/MS (liquid chromatography mass spectrometry). Using these techniques, the limit of detection (LOD) determined was 1.5 μg kg-1 and limit of quantification (LOQ) was 0.005 mg kg-1 for all analytes. The residue levels on mango initially were 0.265 and 0.55 mg kg-1 for azoxystrobin, 0.63 and 0.974 mg kg-1 for buprofezin, 0.635 and 0.98 mg kg-1 for dinocap and 0.203 and 0.35 mg kg-1 for hexaconazole from standard and double dose treatments, respectively. The dissipation rate of the pesticides on mango fruits was about the same except for azoxystrobin, which dissipated slowly compared with others. The half-life of degradation (DT50) of azoxystrobin was 10.4-12.1 days; buprofezin, 5.8-8.5 days; dinocap, 5.4-6.2 days; and hexaconazole, 4.4-6.1 days. The pre-harvest interval (PHI) based on European Union (EU) MRL (maximum residue limit) requirements were 1 day for azoxystrobin, 15 and 26 days for buprofezin, 27 and 34 days for dinocap, and 19 and 30 days for hexaconazole. The results of this study can be used to produce mango fruits safe for consumption and to meet the regulatory requirements for export of mango fruits from India.

The Red River Delta is a major agricultural production area of Vietnam with year-round use of pesticides for paddy rice cultivation and other production systems. The delta is protected from flooding, storm surges and saline water intrusion by a sophisticated river and sea-dyke system. Little is known about the effects of such a dyke system on pesticide pollution in the enclosed landscape. Our aim was to address this gap by i) determining pesticide prevalence in soils and sediments within a dyked agricultural area, and by ii) assessing whether and to which degree this dyke system might affect the spatial distribution of pesticides. After sampling paddy rice fields (topsoil) and irrigation ditches (sediment) perpendicular to the dyke in Giao Thuy district, we analysed 12 of the most commonly used pesticides in this area. In soils, we detected most frequently isoprothiolane (100% detection frequency), chlorpyrifos (85%) and propiconazole (41%) while in sediments isoprothiolane (71%) and propiconazole (71%) were most frequently found. Maximum concentrations reached 42.6 μg isoprotiolane kg-1 in soil, and 35.1 μg azoxystrobin kg-1 in sediment. Our results supported the assumption that the dyke system influenced residue distribution of selected pesticides. More polar substances increasingly accumulated in fields closer to the sea-dyke (R2 = 0.92 for chlorpyrifos and 0.51 for isoprothiolane). We can thus support initiatives from local authorities to use the distance to dykes as a mean for deliniating zones of different environmental pollution; yet, the degree at which dykes influence pesticide accumulation appear to be compound specific.

The use of fungicides is common in mushroom cultivation, but no study was carried out applying reduced doses of fungicides in order to increase yield, taking account the hormesis effect. The aim of this manuscript was to verify the effects of different concentrations of fungicides to stimulate the productivity of different strains of Agaricus bisporus. Two stages were developed, an in vitro study to define the best concentration to be applied in the second experiment an agronomic study, which consisted of the application of the selected fungicides, in their respective concentrations, in an experiment carried out in the mushroom chamber. Clearly, the result of the hormesis effect on mushroom cultivation can be verified. The results obtained in the 1st stage of the study (in vitro) were not always reproduced in the 2nd stage of the study (in vivo). The kresoxim methyl active ingredient may be an important chemical agent, while strain ABI 15/01 may be an extremely important biological agent to increase yield in the study of hormesis effects.

A liquid chromatography-electrospray ionization tandem mass spectrometry method was developed for simple and accurate detection of the fungicides difenoconazole, propiconazole and pyraclostrobin in peppers and soil. Three fungicides residues were extracted from samples by acetonitrile and cleaned up by dispersive solid-phase extraction before instrumental analysis. The accuracy and precision of the method were evaluated by conducting an intra- and inter-day recovery experiment. The limits of quantification and detection of difenoconazole, propiconazole and pyraclostrobin in pepper and soil were 0.005 and 0.0015 mg/kg, respectively. The recoveries were investigated by spiking pepper and soil at three levels, and were found to be in the ranges 79.62-103.15% for difenoconazole, 85.94-103.35% for propiconazole and 80.14-97.69% for pyraclostrobin, with relative standard deviations <6.5%. Field experiments were conducted in three locations in China. The half-lives of difenoconazole, propiconazole and pyraclostrobin were 5.3-11.5 days in peppers and 6.1-32.5 days in soil. At harvest, pepper samples were found to contain difenoconazole, propiconazole and pyraclostrobin well below the maximum residue limits of European Union at the interval of 21 days after last application following the recommended dosage.