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Data on the natural occurrence of Alternariamycotoxins in apple by-products is lacking in Argentina and the risk of exposure to these mycotoxins has not been characterized before. The levels of alternariol (AOH), alternariol monomethyl ether (AME), altenuene (ALT), tenuazonic acid (TeA), tentoxin (TEN), altertoxin-I (ATX-I), altertoxin-II (ATX-II), alternariol 3-sulfate (AOH-3-S), alternariol 3-glucoside (AOH-3-G), alternariol monomethyl ether 3-sulfate (AME-3-S), and alternariol monomethyl ether 3-glucoside (AME-3-G) were determined in clarified and cloudy apple juices, marmalades, and apple-based infant food from the Argentinean market, and the risk of exposure was characterized. Detectable levels of AME, TEN, TeA, AME-3-S and AOH-3-G were found in clarified juices, while the same mycotoxins plus AOH were found in cloudy apple juices in higher concentrations. AME, TEN, TeA and AOH-3G were detected in marmalades, and AOH, AME, TEN and TeA in apple infant food. Probabilistic exposure assessment and risk characterization were carried out for children between 6 months and 5 years old in Argentina. The highest risk of exposure affected children between 6 and 23 months from the consumption of apple infant food and mainly associated with the alternariols. Better control strategies to prevent the incorporation of Alternariamouldy core into the process line and the establishment of legislation for Alternariamycotoxins are needed in Argentina.

Juveniles are considered as one of the most vulnerable population groups concerning mycotoxins and their modified forms. The weaning stage is a particularly vulnerable period in the life of mammals, reflected in intestinal and immune dysfunction. The current study investigated the toxicokinetic (TK) characteristics of zearalenone (ZEN), zearalenone-14-glucoside (ZEN14G), and zearalenone-14-sulfate (ZEN14S) in weaned (4-week-old) piglets, by means of oral and intravenous administration of equimolar doses, i.e., 331, 500, and 415 μg/kg bodyweight, respectively. Plasma and urine were sampled pre- and post-administration and were quantitatively analyzed for ZEN, ZEN14G, ZEN14S, and in vivometabolites by liquid chromatography–high-resolution mass spectrometry. Tailor-made TK models were elaborated to process data. A statistical comparison of the results was performed with TK data obtained in a previously reported study in pigs of 8 weeks of age. Additionally, porcine plasma protein binding was determined to support TK findings. The TK results for ZEN, ZEN14G, and ZEN14S, obtained in 4- and 8-week-old pigs, revealed significant age-related differences, based on differences in intestinal permeability, body fat content, gastrointestinal transit time, and biotransformation, with a special emphasis on an increased absorbed fraction of ZEN14G, i.e., 94 vs 61% in 4- compared to 8-week-old pigs. Since the growing pig has been reported to be a suitable pediatric animal model for humans concerning TK processes, these results may contribute to refine the risk assessment concerning modified ZEN forms in juvenile animals and humans.

A critical point for the successful development of a fluorescent quantum dot (QD)-based immunoassay is maintaining the high fluorescence quantum yield of QDs during hydrophilization and bioconjugation. In this paper, we carefully designed CdSe/CdS and CdSe/CdS/ZnS core–shell heterostructures and extended them with silica coating of different surface composition allowing preservation of fluorescence quantum yield as high as 70% in aqueous media. The silanized QDs containing epoxy and carboxy surface groups were bioconjugated with monoclonal antibodies. The synthesized fluorescent conjugates were used in a multicolor lateral flow immunoassay for simultaneous determination of two mycotoxins. Zearalenone and deoxynivalenol were chosen as a proof of concept. Cutoff levels for the zearalenone and deoxynivalenol detection were adjusted to be at 40 and 400 μg kg–1, respectively, complying with the European Commission regulation. Validation of the developed test was performed by analysis of 34 naturally contaminated maize and wheat samples; as a confirmatory method, LC–MS/MS was used.

: Mycotoxin exposure during pregnancy has been associated with adverse birth outcomes and poor infant growth. We assessed multiple biomarkers and metabolites of exposure to mycotoxins at birth and their associations with birth outcomes and infant growth in 274 newborns in rural Burkina Faso.

An easy-to-operate membrane-based flow-through test for multiplex screening of four mycotoxins (zearalenone, deoxynivalenol, aflatoxin B1, and ochratoxin A) in a variety of cereal-based feed ingredients and compound feeds, such as wheat, barley, soybean, wheat bran, rice, rice bran, maize, rapeseed meal, and sunflower meal, and various types of complete feed (duckling feed, swine feed, broiler feed, piglet feed) was developed and validated. First, the antibodies were evaluated by enzyme-linked immunosorbent assay and then employed in the membrane rapid test. The cutoff levels for zearalenone, deoxynivalenol, aflatoxin B1, and ochratoxin A were 50, 200, 1, and 10 μg/kg, respectively, based on European regulations and consumers’ requirements. As sample pretreatment, consecutive steps of extraction, dilution, solid-phase extraction by addition of C18 sorbent, and final filtration of supernatant were followed. Both the sample preparation and the analysis procedure were simple, cost-effective, and easy to perform on-site in a nonlaboratory environment. The impact of sample processing on the result of the experiment was investigated supported by experimental design. The validation procedure was performed on the basis of Commission Regulation 2006/401/EC. The numbers of false-positive and false-negative outcomes were <5%, going along with the Commission Decision 2002/657/EC. Liquid chromatography–tandem mass spectrometry was performed as a confirmatory technique.

Mycotoxin exposure during pregnancy has been associated with adverse birth outcomes and poor infant growth in low- and middle-income countries. We assessed multiple biomarkers and metabolites of exposure to mycotoxins during pregnancy and their associations with birth outcomes and infant growth in 305 pregnant participants, between 30 and 34 completed weeks of gestation, in rural Burkina Faso. In this study, whole blood microsamples were analyzed for mycotoxin concentrations using ultra-performance liquid chromatography coupled to tandem mass spectrometry. Unadjusted and adjusted associations between mycotoxin exposure, and birth outcomes and infant growth at 6 months were estimated using linear regression models for continuous outcomes and linear probability models with robust variance estimation for binary outcomes. Infant growth trajectories from birth to 6 months were compared by exposure status using mixed-effects models with random intercept for the individual infant and random slope for the infant’s age. Ochratoxin A (OTA) exposure was detected in 50.8% of the study participants, with aflatoxin G1, aflatoxin M1, cyclopiazonic acid, deoxynivalenol and T-2-toxin being detected in the range of 0.33% and 2.31% of the population. We found no statistically significant ( p≥ 0.05) associations between OTA exposure, and birth outcomes and infant growth. Despite this, the findings indicate a significant presence of ochratoxin A among pregnant participants. Public health policies and nutrition-sensitive interventions must ensure that OTA exposure is reduced in Burkina Faso.

A multiplex lateral flow immunoassay (LFIA) for the determination of the mycotoxins deoxynivalenol, zearalenone, and T2/HT2-toxin in barley was developed with luminescent quantum dots (QDs) as label. The synthesized QDs were hydrophilized by two strategies, that is, coating with an amphiphilic polymer or silica. The water-soluble QDs were compared with regard to their bioconjugation with monoclonal antibody (mAb) and were tested on a LFIA. Silica-coated QDs that contained epoxy groups were most promising. Therefore, green, orange, and red epoxy-functionalized silica-coated QDs were conjugated with anti-ZEN, anti-DON, and anti-T2 mAb, respectively. The LFIA was developed in accordance with the European Commission legal limits with cutoff limits of 1000, 80, and 80 μg/kg for deoxynivalenol, zearalenone, and T2/HT2-toxin, respectively. The LFIA gave a fast result (15 min) with a low false-negative rate (<5%), and the results were easy to interpret without any sophisticated equipment.

To explore differences of zearalenone (ZEN) metabolism between various species, phase I and II metabolism by liver microsomes of animals and human were investigated using ultra high-pressure liquid chromatography-quadrupole/time-of-flight mass spectrometry (UHPLC-Q/TOF MS). A total of 24 metabolites were identified, among which 12 were reported for the first time. Reduction, hydroxylation, and glucuronidation were the major metabolic pathways of ZEN, and significant differences in various species were also observed. Reduction was the main reaction in swine and human, whereas hydroxylation was predominant in rats, chickens, goats, and cows in in vitro systems. Furthemore, in vivo metabolism of ZEN in rats and chickens was investigated, and 23 and 6 metabolites were identified in each species, respectively. Reduction, hydroxylation, and glucuronidation were the major metabolic pathways in rats, while reduction and sulfation predominated in chickens. These results further enrich the biotransformation profile of ZEN, providing a helpful reference for assessing the risks to animals and humans.

After being incubated with animal and human liver microsomes, metabolites of phase I and II were investigated. A comparison was performed by ultrahigh performance liquid chromatography-quadrupole/time-of-flight coupled to mass spectrometry (UHPLC-Q/TOF). Consequently, a total of four phase I metabolites and three glucuronide binding metabolites of T-2 toxin were discovered. Although a significant metabolic difference was observed among six species, HT-2 toxin was the major product in all species. In addition, the in vivo metabolism of T-2 toxin after oral administration was also investigated in chickens, In total, 18 metabolites were detected, of which 13 were novel, to our knowledge, and reported for the first time. To elucidate the structures of these metabolites, besides accurate mass data from their MS and MS2spectra, online hydrogen/deuterium (H/D) exchange technique was also carried out. These new metabolites were regarded as 3′-hydroxy-T-2 3-sulfate, 3′-hydroxy-HT-2 3-sulfate, 4′-hydroxy-HT-2, 3′,4′-dihydroxy-HT-2, 4′-carboxyl-T-2, 4′-carboxyl-HT-2, 4′-carboxyl-4′-hydroxy-T-2, and their isomers, implying that T-2 toxin was metabolized more extensively in animals than previously thought. Furthermore, 3′-hydroxy-HT-2, 4′-carboxyl-T-2, 3′-hydroxy-T-2, HT-2 toxin, and neosolaniol were identified to be the major metabolites of T-2 toxin in chickens. The present study expands existing knowledge about T-2 toxin metabolism, informing assessments of the impact T-2 toxin exposure and metabolism on health.

Beauvericin (BEA) and enniatins are toxic ionophoric cyclodepsipeptides that mainly occur in grains. As such, their presence in food commodities poses a concern for public health. To date, despite recent European Food Safety Authority emphasis on the need for more data to evaluate long-term toxicity effects, no suitable affinity reagents are available to detect the presence of BEA and derivatives in food samples. We here report on the synthesis of a small library of artificial receptors with varying cavity sizes and different hydrophobic building blocks. Immobilization of one of the receptors on solid support resulted in a strong retention of beauvericin, thus revealing promising properties as solid-phase extraction material for sample pretreatment. Furthermore, treatment of HepG2 cells with the most promising receptor markedly reduced beauvericin-induced cytotoxicity, hinting toward the possibility of using synthetic receptors as antidotes against ionophoric toxins.

A multiplex immunochromatographic assay (ICA) based on dual-color fluorescent microspheres (FMs) as a sensitive label was developed for the first time. Two typical algae toxins, microcystin-LR (MC-LR) and okadaic acid (OA), were chosen as proof-of concept targets to evaluate the feasibility of this ICA format. Commercial red- and green-colored FMs were selected to couple with monoclonal antibodies as fluorescent probes. The use of dual-wavelength FMs as labels guaranteed a lower consumption of material strips, lower sample volume, and shorter reaction time without increasing the length of ICA strips. Under optimal conditions, the multiplex FM-ICA could be completed in 20 min and reached limits of detection for the simultaneous determination of MC-LR and OA in fish samples, which were 0.074 and 2.42 μg/kg, respectively. The developed technique was validated using artificially spiked and naturally contaminated fish samples. Ultra-high-performance liquid chromatography-tandem mass spectrometry was used as confirmatory technique. In summary, this portable ICAs detection mode based on dual-wavelength FMs provided a reliable and sensitive on-sitedetection of multiple contaminants in food samples, which opens a new field for application of FMs in food safety.

The aim of this study was to determine the toxicokinetic characteristics of ZEN and its modified forms, α-zearalenol (α-ZEL), β-zearalenol (β-ZEL), zearalenone-14-glucoside (ZEN14G), and zearalenone-14-sulfate (ZEN14S), including presystemic and systemic hydrolysis in pigs. Crossover pig trials were performed by means of intravenous and oral administration of ZEN and its modified forms. Systemic plasma concentrations of the administered toxins and their metabolites were quantified and further processed via tailor-made compartmental toxicokinetic models. Furthermore, portal plasma was analyzed to unravel the site of hydrolysis, and urine samples were analyzed to determine urinary excretion. Results demonstrate complete presystemic hydrolysis of ZEN14G and ZEN14S to ZEN and high oral bioavailability for all administered compounds, with further extensive first-pass glucuronidation. Conclusively, the modified-ZEN forms α-ZEL, β-ZEL, ZEN14G, and ZEN14S contribute to overall ZEN systemic toxicity in pigs and should be taken into account for risk assessment.

The synthesis of the first synthetic receptor showing high affinity for the toxic ionophoric cyclodepsipeptide beauvericin is described. Binding results in a pronounced increase in fluorescence intensity of the receptor, while this increase is not observed for a very similar ionophore such as valinomycin. Experiments that shed light on the nature of this selectivity are discussed.

The lack of information on HT-2 toxin leads to inaccurate hazard evaluations. In the present study, toxicokinetic studies of HT-2 toxin were investigated following intravenous (iv) and oral administration to rats at dosages of 1.0 mg per kilogram of body weight. After oral administration, HT-2 toxin was not detected in plasma, whereas its hydroxylated metabolite, 3′-OH HT-2 was identified. Following iv administration, HT-2 toxin; its 3′-hydroxylated product; and its glucuronide derivative, 3-GlcA HT-2, were observed in plasma, and the glucuronide conjugate was the predominant metabolite. To explore the missing HT-2 toxin in plasma, metabolic studies of HT-2 toxin in liver microsomes were conducted. Consequently, eight phase I and three phase II metabolites were identified. Hydroxylation, hydrolysis, and glucuronidation were the main metabolic pathways, among which hydroxylation was the predominant one, mediated by 3A4, a cytochrome P450 enzyme. Additionally, significant interspecies metabolic differences were observed.

A monoclonal antibody (mAb) (2B10) against zearalenone (ZEN) was prepared for the first time in this study. The mAb was class specific and showed no cross reactivity to other groups of mycotoxins. A competitive microarray assay based on a novel solid supporting material, an integrated poly(dimethylsiloxane) (iPDMS), is proposed for qualitative and/or semiquantitative determination of ZEN in cereal samples. The limit of detection (LOD) and limit of quantification (LOQ) of the iPDMS based method were 0.53 μg kg−1and 1.02 μg kg−1for ZEN, respectively, which was lower than the EU maximum limit (ML). Furthermore, recoveries ranged from 93% to 114% and the coefficients of variation were below 15%. The developed method was successfully applied to analyse 29 cereal samples and the results showed that three corn flour samples were ZEN positive, and two corn flour samples were determined to be suspicious, which was in a good agreement with the results obtained using LC-MS/MS. It is worth noting that, compared with ELISA, the developed microarray assay is rapid, simple and does not require a blocking step. In addition, owing to its advantage of zero background, appropriately increasing the concentration of coating antigen and horseradish peroxidase (HRP)-labeled secondary antibody can enhance the sensitivity of the membrane and reduce the matrix effects of the sample.

In this work, there is a detailed description of the whole process of biocompatible CIS/ZnS QDs production. Special attention was paid to the stability of QDs against photooxidation. It was shown that Cu/In ratio greatly affected not only nanocrystals PLQYs but photostability as well. CIS/ZnS QDs with Cu/In = 1:4 ratio showed high photostability under UV illumination both in toluene and aqueous solutions. Meanwhile, photoluminescence of CIS/ZnS QDs with Cu/In = 1:1 ratio was completely quenched after several hours under UV illumination, though their initial QY was as high as 40% with peak maximum at 740 nm. QDs were transferred to water by polymer encapsulation and were subsequently modified with polyethers Jeffamines, cheap analogues of PEG-derivatives. Three types of hydrophilic QDs differing in size, PEG content, and surface charge were obtained for further investigation and comparison of their cytotoxicity and hemocompatibility. It was shown that both leucocytes size distribution and coagulation activation change after introduction of polyethers into QDs polymeric shell, while red blood cells and platelets size distribution as well as hemolysis rate did not show any different results among different QDs and the polymer itself. All three types of QDs showed only slight cytotoxicity. Confocal microscopy proves penetration of hydrophilic CIS/ZnS QDs inside cells, so the low QDs cytotoxocity cannot be explained by low cellular uptake of the QDs and indicated low QDs toxicity in general.

Estrogen receptor β (ERβ) and NOD-like receptor family pyrin domain containing 6 (NLRP6) are highly expressed in intestinal tissues. Loss of ERβ and NLRP6 exacerbate colitis in mouse models; however, the underlying mechanisms are incompletely understood. Here, we report that ERβ directly activates the NLRP6 gene expression via binding to estrogen responsive element of Nlrp6 gene promoter. ERβ also physically interacts with the NLRP6 nucleotide-binding domain and promotes NLRP6 inflammasome assembly. The ERβ-NLRP6 axis then interacts with multiple autophagy-related proteins, including ULK1, BECN1, ATG16L1, LC3B, and p62, and affects the autophagosome biogenesis and autophagic flux. Finally, NLRP6-mediated autophagy suppresses the inflammatory response by promoting the K48-linked polyubiquitination of ASC, Casp-1 p20, IL-1β, TNF-α, and prohibitin-2. Thus, ERβ-NLRP6 direct an anti-inflammatory response by promoting autophagy. Our work uncovers an ERβ-NLRP6-autophagy pathway as a regulatory mechanism that maintains intestinal epithelial cell homeostasis and facilitates tissue repair in colitis.

Food safety monitoring and toxicological research of mycotoxins are still in need of large quantities of high-purity deoxynivalenol (DON). To attain this purpose, a rapid, economical and reproducible purification method was developed for large-scale production of DON from rice culture inoculated with a DON-producing Fusarium graminearumstrain JYH. The inoculated rice culture was first extracted with acetonitrile–water (84/16, v/v). The extracts were evaporated to dryness on a rotary evaporator after ethyl acetate partitioning and then dissolved in water followed by the final purification procedure through preparative high performance liquid chromatography and montmorillonite treatment. A combined approach of ultraviolet spectrometry (UV), ultra high performance liquid chromatography (UHPLC), mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy was applied for the multi-dimensional characterization of the target compound. As a result, the recovery of DON from the crude extract to the final product was up to 70%. An amount of 150 mg DON with the desirable purity of 98.93% could be obtained from 100 g of rice culture, which possessed identical immunochemical characteristics compared to a certified commercial DON standard. This proposed strategy might act as a valuable reference to obtain rather expensive compounds in a straightforward way.

An isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with a fast sample preparation using homemade clean-up cartridges was developed for simultaneous determination of co-occurring mycotoxins exemplified with aflatoxin B1 (AFB1) and T-2 toxin (T-2) in representative biomatrices of rat plasma, heart, liver, kidney, spleen, lung and brain in a total run time of 7 min. The established approach using stable internal standards of [13C17]-AFB1 and [13C24]-T-2 was extensively validated by determining the specificity, linearity (R2≥ 0.9990), sensitivity (lower limit of quantitation at 0.05 ng mL−1), accuracy (70.9–107.7%), precision (RSD ≤ 14.2%) and stability (≥70.8%). Based on this methodological advance, the subsequent kinetics and tissue distribution after oral administration of 0.5 mg kg−1b.w. of both AFB1 and T-2 in rats were thoroughly studied. As revealed, both AFB1 and T-2 were rapidly eliminated with the half-life time (t1/2) in plasma of 8.44 ± 4.02 h and 8.12 ± 4.05 h, respectively. Moreover, AFB1 accumulated in all organs where the highest concentration was observed in liver (1.34 μg kg−1), followed by kidney (0.76 μg kg−1). Notably, only low levels of T-2 were observed in spleen (0.70 μg kg−1) and in liver (0.15 μg kg−1). The achieved data as supporting evidence would substantially promote the practical application of the proposed LC-MS/MS method for in vivotoxicokinetics and toxicity studies of co-occurring mycotoxins imitating natural incidence in rat system.

Hydrogen peroxide induced by the fungicide prothioconazole triggers deoxynivalenol (DON) production by Fusarium graminearumFusariumhead blight is a very important disease of small grain cereals with F. graminearumas one of the most important causal agents. It not only causes reduction in yield and quality but from a human and animal healthcare point of view, it produces mycotoxins such as deoxynivalenol (DON) which can accumulate to toxic levels. Little is known about external triggers influencing DON production. In the present work, a combined in vivo/in vitroapproach was used to test the effect of sub lethal fungicide treatments on DON production. Using a dilution series of prothioconazole, azoxystrobin and prothioconazole + fluoxastrobin, we demonstrated that sub lethal doses of prothioconazole coincide with an increase in DON production 48 h after fungicide treatment. In an artificial infection trial using wheat plants, the in vitroresults of increased DON levels upon sub lethal prothioconazole application were confirmed illustrating the significance of these results from a practical point of view. In addition, further in vitroexperiments revealed a timely hyperinduction of H2O2production as fast as 4h after amending cultures with prothioconazole. When applying H2O2directly to germinating conidia, a similar induction of DON-production by F. graminearumwas observed. The effect of sub lethal prothioconazole concentrations on DON production completely disappeared when applying catalase together with the fungicide. These cumulative results suggest that H2O2induced by sub lethal doses of the triazole fungicide prothioconazole acts as a trigger of DON biosynthesis. In a broader framework, this work clearly shows that DON production by the plant pathogen F. graminearumis the result of the interaction of fungal genomics and external environmental triggers.

A gel-based immunoassay that can be used for the detection of 2,4,6-trinitrotoluene (TNT) in water samples was developed. Four polyclonal antibodies were generated in chickens using TNT derivatives. The assay was based on the immunoaffinity preconcentration and immuno-enzyme analysis of TNT in the gel. The results of the assay, assessed by color development, were evaluated visually and also by using a flatbed scanner and subsequent digital processing of the scanned gel. The most sensitive color mode, parameter S (saturation, HSB mode), was used for the immunoassay optimization and evaluation of the results. The immunoassays with the best parameters were optimized and characterized. A cut-off level of 5 µg TNT L−1was reached for water samples. It was shown that tap and environmental water samples could be analyzed directly, without sample preparation and dilution. The developed test is acceptable for use in an on-site field test to provide rapid (about 15 min for six samples), qualitative and reliable results for making environmental decisions such as identifying “hot spots”, monitoring of military and terrorist activities, and selecting of site samples for laboratory analysis.

Polycyclic aromatic hydrocarbons (PAHs) represent health and environmental concerns. The present study was designed to develop a highly sensitive and reliable immunochemically-based non-instrumental field assay to monitor the presence of benzo[α]pyrene (BAP) in aqueous samples at the ppt-level using visible test evaluation. The corresponding gel-based immunoassay was set up to combine preconcentration and detection of the target analyte using anti-PAH antibodies and horseradish-BAP tracer conjugate in one single cartridge. Water sample preparation, such as extraction, centrifugation, or filtering was found to be unnecessary. No interference by higher water-soluble PAHs at 4000-fold excess compared to BAP was observed. The assay was configured as a qualitative test (positive/negative) at the cut-off level of 5 ng L-1, however, this level can be adapted to the required analyte concentration by rather simple adjustment of the anti-PAH antibody concentration of the test zone. Test validation was performed with real samples such as surface water, melted snow, and tap water using high performance liquid chromatography with fluorescent detection (HPLC-FLD) with solid phase extraction for validation.

A rapid antibody-based assay for the detection of ochratoxin A in cocoa powder is described, involving sequential clean-up and visual detection of the toxin (“clean-up tandem assay column”). The screening test was developed to have a cut-off level of 2 µg kg−1and was shown to have false positive and false negative rates of 10 and 2%, respectively. Analysis of six samples can be carried out in the field in approximately 30 min by untrained workers. Using the proposed rapid screening test, 10 retail cocoa powders were found to contain no detectable levels of ochratoxin A (<2 µg kg−1). These samples were also found to be negative (<2 µg kg−1) when analysed using an LC–MS/MS method.

Grains (cereals and legumes) contribute significantly to in achieving food and nutritional security. However, these commodities are susceptible to fungal contamination, accompanied by the production of toxic secondary metabolites known as mycotoxins. Despite existence of extensive literature concerning prevention and control of mycotoxin contamination in these commodities, its packaging into guidelines or advice remains complex for non-scientists to comprehend thus restricting effective implementation of these measures. In an effort to ease comprehension and implementation of the mycotoxin prevention and control in grains, this paper synthesizes literature on the subject and proposes a framework of five keys with: (1) Recommendations (what); (2) Actionable steps/instructions to achieve recommendations (how) and, (3) Reasoning behind the recommendations (why). This concept of the five keys was originally written from a developing country perspective. However, most of these measures could also be applicable in a global context.

A plethora of scientific researches has constantly studied the electrochemical detection of different pesticides due to their serious effects on humans. Imidacloprid (IMD) is a pesticide that belongs to the neonicotinoids group. In the current work, a carbon paste electrode (CPE) modified with bismuth ferrite microflowers (BiFeO3/CPE) was used for sensing of IMD in water samples in the presence of fipronil (FIP). The modifier (BiFeO3) was characterized by different techniques. To evaluate the efficiency of the developed electrode towards the detection of both IMD and FIP in water samples, the square wave voltammetry (SWV) technique was employed and spiked water samples were used. The calibration curves were within the range of 1.0–100.0 μM for both analytes. The detection limits (LODs) for IMD and FIP were determined as 0.97 μM and 0.81 μM, respectively. The recovery percentages for IMD and FIP in environmental water samples varied from 90.0%–105.0%. In conclusion, the developed sensor was successfully optimized and can be used for the simultaneous detection of both IMD and FIP in environmental water samples.