Health concerns associated to tropane alkaloids in maize food products in Serbia.

Following the implementation of the European regulation limiting the presence of tropane alkaloids in certain foods, a survey was conducted in Serbia on 103 maize products (grits, polenta and semolina) to determine atropine and scopolamine content using liquid chromatography with tandem-mass spectrometry analysis (LC-MS/MS). The probability of exceeding the Acute Reference Dose (ARfD; 0.016 µg/kg bw/day) of the sum of atropine and scopolamine by consuming these products was tested. Overall, across age categories - children, younger and older adolescents, and adults, the group ARfD was exceeded by 21.4%, 17.5%, 11.7% and 11.7% of the samples, with maximum exposure reaching as much as 19-, 13-, 9- and 9-fold the group ARfD, respectively. Nevertheless, polenta could be the most favorable dietary option (17.9% of positive samples, 7.7% resulting in excessive exposure in children, reaching a maximum of 1.4-fold the group ARfD). According to the reported findings, adverse health effects of tropane alkaloids cannot be ruled out. The Margin of Exposure, founded on a clinically significant acute effects dose established by FAO/WHO, ranged from 1194 to 2381 (mean) and from 28 to 56 (95th percentile) across age categories. These estimates should certainly draw the attention of food authorities and nutritionist, particularly in the case of highly sensitive populations with contraindications and high consumers of corn products, such as coeliac patients.

Microbial Resistance to Antibiotics and Biofilm Formation of Bacterial Isolates from Different Carp Species and Risk Assessment for Public Health.

The aim of this research was to investigate the effects of biofilm on antibiotic resistance of the bacterial isolates present in fish meat and to assess the risk of antibiotic residues for public health. Common carp, silver carp and grass carp fishes were purchased from retail stores for an in vitro biofilm investigation and a drug-resistant pattern determination. In all samples, up to 104 CFU/g of bacteria, such as Escherichia coli, Aeromonas hydrophila, Shewanella putrefaciens, Vibrio spp. and Staphylococcus spp., were observed. Isolates from the samples and their biofilms were subjected to an antibiogram assay using antibiotics such as amoxicillin, ampicillin, cefotaxime, ciprofloxacin, chloramphenicol, gentamicin, streptomycin, tetracycline and trimethoprim. Obtained results showed that some of the isolates were sensitive to antibiotics and some were resistant. Results of LC-MS/MS analysis showed that antibiotics residues were present in fish samples in the range between 4.9 and 199.4 µg/kg, with a total sum of 417.1 µg/kg. Estimated daily intake (EDI) was established to be 0.274 µg/kg of body weight/day for men and 0.332 µg/kg of body weight/day for women, with an acceptable daily intake (ADI) of 8.5 and 7.0 µg/kg of body weight/day for men and women, respectively. The results of the present study, therefore, highlight the safe consumption of fresh fish.

Occurrence and Determination of Alternaria Mycotoxins Alternariol, Alternariol Monomethyl Ether, and Tentoxin in Wheat Grains by QuEChERS Method.

The Alternaria mycotoxins such as alternariol (AOH), alternariol monomethyl ether (AME), and tentoxin (TEN) are mycotoxins, which can contaminate cereal-based raw materials. Today, wheat is one of the most important crops in temperate zones, and it is in increasing demand in the Western Balkans countries that are urbanizing and industrializing. This research aimed to investigate the occurrence and determine the concentration of Alternaria mycotoxins AOH, AME, and TEN in wheat samples from the Republic of Serbia and the Republic of Albania, harvested in the year 2020 in the period between 15 June and 15 July. A total of 80 wheat grain samples, 40 from each country, were analyzed by an QuEChERS (quick, easy, cheap, effective, rugged, and safe) method. From the obtained results, it can be seen that the mean concentration of AOH was 3.3 µg/kg and AME was 2.2 µg/kg in wheat samples from Serbia, while TEN from both Serbia and Albania was under the limit of quantification (

Atropine and Scopolamine in Maize Products from the Retail Stores in the Republic of Serbia.

The cereal grains, which represent the cultivated grasses fruits, supply almost half of the total caloric requirements for humans and provide more nourishment compared with any other class of the food. Out of many cereals used for food, maize, rice, and wheat are the most important food resources for humans, representing 94% of the total cereals consumption. According to the data of the Republic Institute of Statistics for the year 2018, the harvested areas of corn amount to 906,753 hectares. The production of about 7 million tons was achieved with an average yield of 7.7 t/ha according to the Ministry of Agriculture of the Republic of Serbia. Serbia is still among the ten largest exporters of wheat and corn in the world for the period of 2014/15-2017/18. More precisely, it ranks seventh in the export of corn. Utilization of maize products for food animal nutrition (1000 t) is 491,48, and for industrial processing (1000 t) 278,862 expressed as the total consumption (1000 t) is 769,910. Therefore, a total of 103 samples of maize products were analyzed for the presence of toxins, i.e., tropane alkaloids (TAs). The samples were collected from the retail stores in the Republic of Serbia in 2021 and analyzed for the presence of atropine and scopolamine (33 corn grits, 39 polenta, and 31 semolina samples). Therefore, the Recommendation 2015/976/EU on the monitoring of TAs in food was adopted by the EU Commission to obtain more occurrence data on TAs in food. The monitoring extent, however, is restricted because reliable analytical methods and appropriate sensitivity are limited. There was a limit of 1 g/kg for each atropine and scopolamine in cereals containing millet, sorghum, buckwheat, or their derivatives. All the samples were analyzed by the LC-MS/MS. The LOQ was set at 1.0 µg/kg. Out of the total 103 tested samples, 32 samples (31.1%) were contaminated with atropine and scopolamine in concentrations above the LOQ. The highest concentrations of the studied TAs were observed in a semolina sample-atropine: 58.80 µg/kg, scopolamine: 10.20 µg/kg. The obtained results indicate that the TAs concentrations are above the LOQ which can be considered potential human and animal health hazards.

Chemical Composition of Ambrosia trifida L. and Its Allelopathic Influence on Crops.

Phytotoxic substances released by invasive plants have been reported to have anti-pathogen, anti-herbivore, and allelopathic activity. The aim of this study was to determine the allelopathic influence of the Ambrosia trifida L. on oxidative stress parameters (the lipid peroxidation process; reduced glutathione (GSH) content; and activity of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), and peroxidase (PX)) and phenolic compounds (total phenolic and tannin content) in maize (Zea mays L.), soybean (Glycine max L.), and sunflower (Helianthus annuus L.) crops to explore the effect of released allelochemicals through A. trifida root on crops. An analysis by HPLC confirmed the presence of protocatechuic acid, p-hydroxybenzoic acid, vanillic acid, and syringic acid as major components in the A. trifida. Based on the obtained results for oxidative stress parameters, it can be concluded that the sunflower was the most sensitive species to A. trifida allelochemicals among the tested crops. The other two crops tested showed a different sensitivity to A. trifida. The soybean did not show sensitivity, while the maize showed sensitivity only 10 days after the sowing.

Pesticide active substances in infant food in Serbia and risk assessment.

Risks posed by pesticide residues in infant food urge for protection of the most vulnerable part of our population. In the current study a total of 54 samples of infant food (juice and purée) were collected on the Serbian market. Liquid chromatography-tandem mass spectrometry with electrospray ionization method detected 18 out of 69 analyzed pesticide active substances in 55.6% of the samples, most frequently carbendazim and acetamiprid. Domestic products as opposed to the imported ones showed a substantially higher proportion of positive (85% vs 38%) and noncompliant (10% vs 0%) samples, a number of pesticides detected (15 vs 8), the proportion of the samples with multiple residues (85% vs 15%), the maximum number of residues in an individual sample (7 vs 2). Risk assessment was performed for the present pesticide active substances, which was estimated to remain below the level of concern for both acute and chronic adverse health effects.

Influence of Different Tetracycline Antimicrobial Therapy of Mycoplasma (Mycoplasma synoviae) in Laying Hens Compared to Tea Tree Essential Oil on Table Egg Quality and Antibiotic Residues.

The food of animal origin that is the most consumed is the table egg, but laying hens treated with antibiotics can produce eggs contaminated with antibiotic residues. Residues of antibiotics may present a risk for consumer health. Keeping in mind that laying hens almost always suffer from Mycoplasma (Mycoplasma synoviae), for which they are treated with antibiotics, high-quality egg production is even harder. Our research aimed to investigate the influence of three different antibiotics compared to the tea tree (Melaleuca alternifolia) essential oil administered to naturally infected laying hens with M. synoviae, on antibiotic residues in eggs as well as the egg nutritive and sensory qualities. A total of 20,000 laying hens, housed in one facility and divided into four lines each consisting of 5000 hens naturally infected with M. synoviae, was used. For the antimicrobial therapy, tetracycline (TC), oxytetracycline (OTC) and chlortetracycline (CTC) were used, respectively. As a control, tea tree essential oil (TT) was used. Based on the gained results all tetracyclines treatment residue values were significantly (p < 0.05) higher compared to the control treatment (TT), but without any significant differences (p > 0.05) between themselves. The results showed no differences in the nutritive and the sensory qualities of eggs between the control and the experimental treatments (p > 0.05). Keeping in mind the obtained results from this study, it can be concluded that tea tree essential oil could be successfully used as a natural antibiotic in the treatment of M. synoviae, without any adverse effects on table egg quality.

Isolation, Biochemical and Genomic Characterization of Glyphosate Tolerant Bacteria to Perform Microbe-Assisted Phytoremediation.

The large-scale use of the herbicide glyphosate leads to growing ecotoxicological and human health concerns. Microbe-assisted phytoremediation arises as a good option to remove, contain, or degrade glyphosate from soils and waterbodies, and thus avoid further spreading to non-target areas. To achieve this, availability of plant-colonizing, glyphosate-tolerant and -degrading strains is required and at the same time, it must be linked to plant-microorganism interaction studies focusing on a substantive ability to colonize the roots and degrade or transform the herbicide. In this work, we isolated bacteria from a chronically glyphosate-exposed site in Argentina, evaluated their glyphosate tolerance using the minimum inhibitory concentration assay, their in vitro degradation potential, their plant growth-promotion traits, and performed whole genome sequencing to gain insight into the application of a phytoremediation strategy to remediate glyphosate contaminated agronomic soils. Twenty-four soil and root-associated bacterial strains were isolated. Sixteen could grow using glyphosate as the sole source of phosphorous. As shown in MIC assay, some strains tolerated up to 10000 mg kg-1 of glyphosate. Most of them also demonstrated a diverse spectrum of in vitro plant growth-promotion traits, confirmed in their genome sequences. Two representative isolates were studied for their root colonization. An isolate of Ochrobactrum haematophilum exhibited different colonization patterns in the rhizoplane compared to an isolate of Rhizobium sp. Both strains were able to metabolize almost 50% of the original glyphosate concentration of 50 mg l-1 in 9 days. In a microcosms experiment with Lotus corniculatus L, O. haematophilum performed better than Rhizobium, with 97% of glyphosate transformed after 20 days. The results suggest that L. corniculatus in combination with to O. haematophilum can be adopted for phytoremediation of glyphosate on agricultural soils. An effective strategy is presented of linking the experimental data from the isolation of tolerant bacteria with performing plant-bacteria interaction tests to demonstrate positive effects on the removal of glyphosate from soils.

Phytotoxic effects of irrigation water depending on the presence of organic and inorganic pollutants.

Irrigation is one of the most important uses of surface waters in the agricultural region of Vojvodina province (Serbia). The aim of the study was to assess the quality of water from Stara Tisa meander, based on the levels of pollution with metals, volatile compounds (VOC), pharmaceuticals, pesticides, and pathogenic bacteria, on sunflower, cabbage, cucumber, maize, barley, buckwheat, sorghum, radish, beans, and white mustard. Microbiological analysis was carried out using the dilution method and phytotoxicity assay according to ISTA filter paper method (germination energy (GE), germination (G), root and shoot length, fresh and dry weight). The sample was slightly contaminated with domestic, industrial, and agricultural xenobiotics and had low levels of nitrogen substances, metals, and organic micropollutants. Pesticides, metolachlor, tebuconazole, propiconazole, imidacloprid, and thiametoxam were detected at levels exceeding the maximum admissible concentrations (MACs), i.e., the sum value for neonicotinoids. The number of saprophytic (2.27 × 10(6) CFU mL(-1)) and coliform bacteria (5.33 × 10(2) CFU mL(-1)) was very high. The total number of sulphite reducing clostridia (10 cells mL(-1)) and Escherichia coli (5 cells mL(-1)) was very low. The GE and G of all tested plants, except sunflower, were not influenced by the total chemism of water sample. However, it inhibited root lengths of sunflower, cucumber, maize, and barley and stimulated shoot lengths of all species except maize and white mustard. These results indicate that it can be used for irrigation of cabbage and radish from the chemical point of view, but the microbiological traits should be considered prior to consumption since they are consumed raw. The overall results suggest that water from Stara Tisa should be purified before using for agricultural purposes.

Development and Validation of Chronopotentiometric Method for Imidacloprid Determination in Pesticide Formulations and River Water Samples.

A new electrochemical method for determination of imidacloprid using chronopotentiometry on thin film mercury and glassy carbon electrode was presented. The most important experimental parameters of chronopotentiometry were examined and optimized with respect to imidacloprid analytical signal. Imidacloprid provided well-defined reduction peak in Britton-Robinson buffer on thin film mercury electrode at -1.0 V (versus Ag/AgCl (KCl, 3.5 mol/L)) and on glassy carbon electrode at -1.2 V (versus Ag/AgCl (KCl, 3.5 mol/L)). The reduction time was linearly proportional to concentrations from 0.8 to 30.0 mg/L on thin film mercury electrode and from 7.0 to 70.0 mg/L on glassy carbon electrode. The detection limits were 0.17 mg/L and 0.93 mg/L for thin film mercury and glassy carbon electrode, respectively. The estimation of method precision as a function of repeatability and reproducibility showed relative standard deviations values lower than 3.73%. Recovery values from 97.3 to 98.1% confirmed the accuracy of the proposed method, while the constancy of the transition time with deliberated small changes in the experimental parameters indicated a very good robustness. A minor influence of possible interfering compounds proved good selectivity of the method. Developed method was applied for imidacloprid determination in commercial pesticide formulations and river water samples.