Invitro and in vivo anthelmintic efficacy of peppermint (Mentha x piperita L.) essential oil against gastrointestinal nematodes of sheep.

Nowadays, the exclusive use of commercial anthelmintics for the treatment of gastrointestinal nematode infections in ruminants is less sustainable due to anthelmintic resistance, as well as the problem of drug residues in animal products and the environment. Therefore, an integrated therapeutic approach is needed, including the search for alternatives to synthetic anthelmintic drugs. The aim of this study was to evaluate the possibility of using the essential oil of peppermint (Mentha x piperita L.) in the control of gastrointestinal nematodes in sheep. For this purpose, the in vitro and in vivo anthelmintic efficacy of this oil and the toxic effects on the hosts were examined. In the in vitro egg hatch test, ovicidal activity varied from 21.0-90.3% depending on the concentration of essential oil used (0.0125, 0.025, 0.049, 0.195, 0.781, 3.125, 12.5, and 50 mg/mL). To some extent, anthelmintic efficacy was confirmed in the in vivo fecal egg count reduction test at a mean dose of 150 mg/kg, with an average reduction of nematode eggs of 26.9 and 46.0% at Days 7 and 14 after treatment, respectively. Furthermore, no toxic effects of applied oil were observed on sheep behavior, kidney, or liver function. The main compounds identified by gas chromatography-mass spectrometry analyzes were menthol (32.6%), menthone (22.0%), menthyl-acetate (10.0%), and isomenthone (9.39%). Due to their complex chemical compositions, numerous bioactive ingredients, and natural origin, herbal formulations represent a potentially valuable alternative for the control of gastrointestinal nematodes in sheep. In this context, the results of the present study showed that peppermint essential oil is one of the promising candidates. Further studies should be performed to collect more data on the safety profile of M. piperita EO in treated animals to find the most appropriate formulation for use in field conditions and to test it against resistant gastrointestinal nematode populations.

Sunflower Honey-Evaluation of Quality and Stability during Storage.

Honey's unique qualities should last for several years when properly stored. Therefore, it is up to manufacturers to choose the right shelf life for their product while also considering the product's nature. Physicochemical parameters (water content, electrical conductivity, free acidity, pH, ash, water-insoluble matter, hydroxymethylfurfural (HMF), sugar content and composition, and diastase activity) were analyzed in 24 samples of sunflower honey collected from several localities in Vojvodina, Serbia. Crystallization indices were also calculated. Furthermore, the impact of eighteen months of room temperature storage (22 ± 2 °C) in a dark place on selected physicochemical parameters (water, HMF, diastase activity, pH value, and free acidity) was investigated. The results of the initial test indicated that the tested samples of sunflower honey from Vojvodina is of good quality because the parameters under examination revealed results that were within the legal bounds of both national and European legislations. Eighteen months of storage at room temperature reduced diastase activity by 2 times, increased HMF content by about 17 times, and decreased the pH value of honey from a mean value of 3.66 to 3.56. The water content was relatively stable at 17.01% before storage and 16.29% after storage. The storage of sunflower honey did not have an impact on the free acidity.

Compounding of Liquid and Solid Dose Adjustable Formulations with Pantoprazole: Comparison of Stability, Applicability and Suitability.

Pantoprazole is a model substance that requires dosage form adjustments to meet the needs of all patients. Pediatric pantoprazole formulations in Serbia are mostly compounded as capsules (divided powders), while in Western Europe liquid formulations are more common. The aim of this work was to examine and compare the characteristics of compounded liquid and solid dosage forms of pantoprazole. Three syrup bases were used: a sugar-free vehicle for oral solution (according to USP43-NF38), a vehicle with glucose and hydroxypropyl cellulose (according to the DAC/NRF2018) and a commercially available SyrSpend Alka base. Lactose monohydrate, microcrystalline cellulose and a commercially available capsule filler (excipient II, composition: pregelatinized corn starch, magnesium stearate, micronized silicon dioxide, micronized talc) were used as diluents in the capsule formulations. Pantoprazole concentration was determined by the usage of the HPLC method. Pharmaceutical technological procedures and microbiological stability measurements were performed according to the recommendations of the EP10. Although dose appropriate compounding with pantoprazole is suitable using both liquid vehicles as well as solid formulations, chemical stability is enhanced in solid formulation. Nevertheless, according to our results, if liquid formulation is a pH adjusted syrup, it could be safely kept in a refrigerator for up to 4 weeks. Additionally, liquid formulations could be readily applied, while solid formulation should be mixed with appropriate vehicles with higher pH values.

Anthelmintic Properties of Essential Oils to Control Gastrointestinal Nematodes in Sheep-In Vitro and In Vivo Studies.

Herbal products such as essential oils may play a promising role in the treatment of infections caused by gastrointestinal nematodes (GINs). The aim of this study was to evaluate the in vitro potential of 11 essential oils (EOs) and one binary combination of isolated EO compounds, as well as the in vivo anthelmintic efficacy of two EO formulations. Four GIN genera were identified in the coproculture examination: Haemonchus, Trichostrongylus, Teladorsagia and Chabertia. The in vitro egg hatch test (EHT) was performed at six different concentrations (50, 12.5, 3.125, 0.781, 0.195 and 0.049 mg/mL) for each EO, whereas in the in vivo faecal egg count reduction test (FECRT), each EO sample was diluted in sunflower oil and orally administrated at a dose of 100 mg/kg to the different group of animals. In the EHT, the EOs of Origanum vulgare, Foeniculum vulgare, Satureja montana, Satureja hortensis and two types of Thymus vulgaris were the most effective. The dominant compounds of these EOs were carvacrol, thymol, anethol, p-cymene and γ-terpinene, indicating their importance for the anthelmintic activity. In the FECRT, both T. vulgaris EO type 1 and linalool:estragole combination show an anthelmintic potential with a mean effect on FECR of approximately 25%. The results suggest the possible role of tested EOs as anthelmintic agents in sheep farms, although further in vivo tests are needed.

Organochlorine pesticides and NDL-PCBs in wild boars from flatland region with intensive agricultural activities.

The objective of this study was to analyse the content of OHCs and NDL-PCBs in wild boar population living in densely populated flatland region with intensive agricultural production. A total of 82 liver samples were analysed by GC-MS method, revealing the presence of minimum one residue in the liver of the majority of animals (92.7%). Most commonly detected residues included ∑ HCH (64.6%), ∑endrin (56.1%) and ∑dieldrin (32.3%). Mean concentrations were in the range ∑endrin > ∑dieldrin > ∑HCH > ∑NDL-PCB > ∑DDT > ∑CHL > ∑endosulfan > methoxychlor. The dominant compound within the total content of NDL-PCBs was PCB 153 (76%). The assessment of daily intake and determination of Health Index indicated that in 6.1% of the samples the consumption of one single portion of wild boar liver would be harmful to human health due to the presence of ∑endrin or NDL-PCBs.

A Potential Anthelmintic Phytopharmacological Source of Origanum vulgare (L.) Essential Oil against Gastrointestinal Nematodes of Sheep.

The development of anthelmintic resistance in sheep gastrointestinal nematodes (GINs) requires novel strategies for the sustainable control of these parasites. This study aimed to evaluate the anthelmintic efficacy of the Origanum vulgare (L.) essential oil (EO) against sheep GINs and to evaluate the possibility of its use in control practice. The in vitro egg hatch test was conducted at eight different concentrations (50, 12.5, 3.125, 0.781, 0.195, 0.049, 0.025, and 0.0125 mg/mL) of the tested EO. For the in vivo fecal egg count reduction test, the EO of O. vulgare was administrated orally at a mean single dose of 150 mg/kg to sheep from two farms in Southern Italy, whereby potential toxic effects to the hosts were also evaluated. In the egg hatch test, the inhibition of egg hatchability varied from 71.3% to 93.7%, depending on the concentration used. The high anthelmintic potential was confirmed in the fecal egg count reduction test with an average reduction of nematode eggs in feces of 43.2% and 60.1% on days 7 and 14 after treatment, respectively. In addition, no toxic effects were noticed during the clinical examination of sheep or by observing blood count and liver or kidney function test results. The obtained results suggest the strong activity of the O. vulgare EO against sheep GINs, probably due to a high percentage of carvacrol (76.21%), whereby it can be considered safe for sheep at the dose tested in vivo. Therefore, it is suitable for use in veterinary practice as a part of an integrated strategy for the control of sheep GINs.

Epidemiology of human trichinellosis in Vojvodina province, Serbia, from 2005 to 2016.

Trichinellosis is one of the most important foodborne diseases in the Eastern European countries. The main objective of this study was to investigate the epidemiological patterns of trichinellosis outbreaks that occurred between 2005 and 2016 in Vojvodina, a northern province of the Republic of Serbia. The average incidence was 3.5 per 100,000 inhabitants. A total of 828 people acquired the infection. The disease occurred in all age groups, slightly more often in males, and quite frequently in a severe form considering the high share of hospitalised patients and the fatal outcome rate (41.6 ± 31.1% and 0.4%, respectively). Trichinella spiralis was confirmed as the causative agent in eight outbreaks. The outbreaks usually occurred among family members due to the consumption of pork or traditional pork products from not tested backyard pigs. Veterinary control measures and the education of consumers and farmers should be implemented to control this zoonotic disease.

PAHs in different honeys from Serbia.

Maximum limits for PAHs content in honey and bee products are not set. The objective of this study was to conduct a market survey of honey for the presence of PAH16 with the aim of detecting potential contamination and health risks for consumers. A total number of 61 honey samples produced in the territory of Serbia were examined. The content of PAH16 was measured using a gas chromatography-mass spectrometric method. Large values were found in some samples: 140.6 µg/kg for chrysene, 136.3 µg/kg for benzo[ghi]perylene, 120.1 µg/kg for benzo[a]pyrene, 87.2 µg/kg for benz[a]anthracene and 79.6 µg/kg for benzo[k]fluoranthene. It was established that 6.6% honey samples were not safe for human consumption because of high benzo[a]pyrene and PAH4 content, which greatly exceeded the maximum limits for all food categories. Considering that screening of honey obtained from the market does not allow accurate detection of the contamination origin requires further investigations to identify potential contamination sources.

OPTIMIZATION AND VALIDATION OF HPLC METHOD FOR TETRAMETHRIN DETERMINATION IN HUMAN SHAMPOO FORMULATION.

High-performance liquid chromatography (HPLC) method with diode array detection (DAD) were optimized and validated for separation and determination of tetramethrin in an antiparasitic human shampoo. In order to optimize separation conditions, two different columns, different column oven temperatures, as well as mobile phase composition and ratio, were tested. Best separation was achieved on the Supelcosil TM LC-18- DB column (4.6 x 250 mm), particle size 5 jim, with mobile phase methanol : water (78 : 22, v/v) at a flow rate of 0.8 mL/min and at temperature of 30°C. The detection wavelength of the detector was set at 220 nm. Under the optimum chromatographic conditions, standard calibration curve was measured with good linearity [r2 = 0.9997]. Accuracy of the method defined as a mean recovery of tetramethrin from shampoo matrix was 100.09%. The advantages of this method are that it can easily be used for the routine analysis of drug tetramethrin in pharmaceutical formulas and in all pharmaceutical researches involving tetramethrin.