Seasonal Dynamics and Physiological Age of Ixodid Ticks Collected from Dogs.

In order to reproduce and complete life cycles, ticks have to feed on different hosts, thus participating as vectors and reservoirs in the maintenance and circulation of different pathogens. Since dogs can serve as suitable hosts for numerous tick species, the aims of this study were to determine tick species and their seasonal occurrence on pet dogs and to compare the accuracy of three indices frequently used to calculate engorged female physiological age. Ticks were collected from dogs brought to veterinary clinics. Three indices were analyzed: scutal index, alloscutal/scutal index ratio, and physiological age index. Four tick species were identified: Ixodes ricinus, Dermacentor marginatus, D. reticulatus, and Rhipicephalus sanguineus group, and the last was the most abundant. The highest number of collected ticks was in April, but two species were continuously active throughout the year. The statistical analyses distinguished the physiological age index as more precise because of lower variability. Dog owners usually ignore regular dog anti-tick treatments throughout the year, as they are not aware that ticks could be active during the winter months. Tick surveillance is unquestionably important in order to monitor and prevent the distribution of these vectors and also the diseases they transmit.

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.

Antimicrobial Activity of Selected Essential Oils against Selected Pathogenic Bacteria: In Vitro Study.

The worldwide problem of infectious diseases has appeared in recent years, and antimicrobial agents are crucial in reducing disease emergence. Nevertheless, the development and distribution of multidrug-resistant (MDR) strains in pathogenic bacteria, such as Escherichia coli, Staphylococcus aureus, Salmonella Typhi and Citrobacter koseri, has become a major society health hazard. Essential oils could serve as a promising tool as a natural drug in fighting the problem with these bacteria. The current study aimed to investigate the antimicrobial effectiveness of tea tree (Melaleuca alternifolia (Maiden and Betche) Cheel), rosemary (Rosmarinus officinalis L.), eucalyptus (Eucalyptus obliqua L'Hér.), and lavender (Lavandula angustifolia Mill) essential oils. The antimicrobial properties of essential oils were screened against four pathogenic bacteria, E. coli, S. aureus, S. Tyhpi, and C. koseri, and two reference bacterial strains, while for the testing, the agar well diffusion method was used. Gas chromatography (GC) and gas chromatography-mass spectrometric (GC-MSD) analyses were performed on essential oils. The obtained results showed that M. alternifolia essential oil is the richest in terpinen-4-ol, R. officinalis and E. oblique essential oils in 1,8-cineole, and L. angustifolia essential oil in α-terpinyl acetate. In addition, the main bioactive compounds present in the essential oil of tea tree are rich in α-pinene (18.38%), limonene (7.55%) and γ-terpinene (14.01%). The essential oil of rosemary is rich in α-pinene (8.38%) and limonene (11.86%); eucalyptus essential oil has significant concentrations of α-pinene (12.60%), p-cymene (3.24%), limonene (3.87%), and γ-terpinene (7.37%), while the essential oil of lavender is rich in linalool (10.71%), linalool acetate (9.60%), α-terpinyl acetate (10.93%), and carbitol (13.05%) bioactive compounds, respectively. The obtained results from the in vitro study revealed that most of the essential oils exhibited antimicrobial properties. Among the tested essential oils, tea tree was discovered to demonstrate the strongest antimicrobial activity. The recorded MIC of S. Typhi was 6.2 mg/mL, 3.4 mg/mL of C. koseri, 3.1 mg/mL of E. coli, and 2.7 mg/mL of E. coli ATCC 25922, compared to M. alternifolia. Similarly, only S. aureus ATCC 25923 showed antimicrobial activity towards R. officinalis (1.4 mg/mL), E. oblique (2.9 mg/mL), and L. angustifolia (2.1 mg/mL). Based on the obtained results, it is possible to conclude that tea tree essential oil might be used as an ecological antimicrobial in treating infectious diseases caused by the tested pathogens.

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.

Comparison of QuEChERS with Traditional Sample Preparation Methods in the Determination of Multiclass Pesticides in Soil.

Background: The increased use of pesticides leads to permanent pollution of soil, and there is a need for continuous monitoring of these agrochemicals in soil. Objective: Three methods for the simultaneous determination of 12 pesticides belonging to eight chemical groups in soil samples were tested and compared based on analytical parameters. Methods: The quick, easy, cheap, effective, rugged, and safe (QuEChERS); traditional solid-liquid extraction (SLE); and Soxhlet extraction were used for soil sample preparation, while detection and quantification of pesticides were performed using gas chromatography-mass spectrometry (GC-MS). Results: The tested methods featured good sensitivity, and with the exception for carbofuran (Soxhlet method, LOD = 29 µg/kg), for the rest of the pesticides, the studied LODs were less than 12 µg/kg. Except for simazine and carbofuran, LODs obtained by Soxhlet extraction were lower than values obtained by other two methods, whereas QuEChERS gave lower LODs than the traditional SLE method for all compounds except atrazine and acetochlor. The recoveries obtained applying QuEChERS, traditional SLE, and Soxhlet methods for multiple analyses of soil samples fortified at 10, 75, and 200 µg/kg of each pesticide were in the ranges 54-103, 40-91, and 12-92%, respectively. Except for chlorothalonil, the highest recoveries were obtained by the QuEChERS method. Soxhlet was better than traditional SLE method for chlorothalonil, heptachlor, and aldrin; organophosphorus pesticides (fenitrothion and diazinon) and trifluralin, gave similar recoveries for both methods. All three methods were proven to be repeatable, with RSDs lower than 19%. Conclusions: Although all tested methods showed as satisfactory regarding most analytical parameters, QuEChERS method showed much better results in terms of confidence, indicating that traditional SLE and Soxhlet extraction still need improvements for determination of multiclass pesticides in soil samples.

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.