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Since ancient times, many plant species within the Thymus genus have been used due to their numerous health benefits, such as antimicrobial, anti-inflammatory, antiseptic, or diuretic activity. While many of the species within this genus were well known and described, Thymus pannonicus All. or Pannonian thyme remains relatively unexplored despite its unique chemical composition and activity. T. pannonicus is a small shrub, spread over central and eastern Europe, with a diverse, location-dependent chemical composition. Within this study, the citral chemotype of T. pannonicus was used, which up to our knowledge was found and described only in Northern Serbia. Therefore, in order to explore the composition of T. pannonicus low-polar extracts and to present a preliminary insight into their bioactivity, supercritical carbon dioxide (ScCO 2 ) extraction was applied (with and without pre-treatments), followed by GC-MS analysis. Different ScCO 2 extraction pressures were tested (100, 150, 200, 250, and 300 bar) in order to determine the most suitable conditions for the extraction of T. pannonicus 's low-polar fraction. Additionally, in order to maximize the yield of the extraction, two types of ScCO 2 extraction pre-treatments were investigated (enzymatic and microwave). The highest extraction yield obtained from untreated plant material was 3.01% ( w / w ), and it was obtained at conditions of 150 bar and 40 °C, while the 4% ( v / w ) enzymatic pre-treatment, with the same conditions, provided a yield of 3.89%. For all of the obtained extracts, the GC-MS analysis showed that oxygenated monoterpenes and sesquiterpenes were the two most dominant groups with principal bioactive compounds such as (E)-citral (18.95-38.17%), (Z)-citral (6.68-14.66%), β-bisabolene (8.2-14.4%), and nerol (6.08-9.67%). The extracts that exhibited the highest concentration of principal bioactive compounds ((Z)-citral, and (E)-citral) were further analyzed for anticancer potential, using short- and long-term cell viability observations on liver cancer cells.

In the production of refined sunflower oil, waxes are removed during the winterization stage, and wax crystals are separated through filtration assisted by filtration aids. Commonly used filtration aids in oil refining include perlite and diatomaceous earth. After winterization, a significant amount of filter cake remains as a by-product and is treated as waste. Today, natural cellulose fibers are being promoted as filtration aids. Their advantages are numerous, both in the production process and from an environmental perspective. However, their only disadvantage is their higher cost. Therefore, in this study, 57 filtration cycles during the industrial sunflower oil winterization step using cellulose-based filtration aids were monitored. Different process parameters, including the pressure differential on the filter, the flow rate of filtered oil, constant pressure period, the quantity of filtered oil, filtration time, the quantity of pre-coating and dosing filtration aids, the volume of filtered oil, the concentration of dosing filtration aid, as well as the mass of separated waxes, were observed. Additionally, artificial neural networks were applied to predict process parameters, optimize the process, and, above all, determine the dosage of filtration aids, which will make the process more economical. The optimal filtration process is performed at a pressure differential of 3.3 bar, lasting a total of 39 h, with 32 h at constant pressure, resulting in 322,503 kg of filtered oil and 90.41 kg of waxes. The optimal quantity of cellulose-based filtration aids employed for pre-coat was 80 kg, and for dosing, 375 kg, with an optimal concentration of 0.12% w/w .

Agricultural waste is underutilized, and sometimes burning them has a negative impact on the environment and human health. This research investigates the untapped potential of extracts from maize, wheat and sunflower waste as natural materials for cutaneous, specifically, cosmetic application. The possibility of incorporating lipid and ethanol extracts from wheat, maize, and sunflower into creams was investigated together with their potential contribution to the structural and functional properties of the topical formulations. Results of the physicochemical characterization show that investigated extracts can be successfully incorporated into creams with satisfactory stability. All extracts showed a desirable safety profile and good antimicrobial activity against various microorganisms. Lipid extracts have proven to be promising structural ingredients of the oil phase, contributing to the spreadability, occlusivity, and emollient effect. Ethanol extracts influenced washability and stickiness of the formulation and could be considered as prospective ingredients in self-preserving formulations. The extracts affected the sensory properties of the creams, mainly the smell and color. These results suggest that the extracts from wheat, maize, and sunflower waste could be used as multifunctional natural ingredients for cosmetic formulations which can replace less sustainable raw materials. This also represents a valorization of waste and is in line with broader sustainability goals.

A lack of strategies for the utilization of harvest residues (HRs) has led to serious environmental problems due to an accumulation of these residues or their burning in the field. In this study, wheat and corn HRs were used as feedstock for the production of microcrystalline cellulose (MCC) by treatment with 2-8% sodium hydroxide, 10% hydrogen peroxide and further hydrolysis with 1-2 M hydrochloric acid. The changes in the FT-IR spectra and PXRD diffractograms after chemical treatment confirmed the removal of most of the lignin, hemicellulose and amorphous fraction of cellulose. A higher degree of crystallinity was observed for MCC obtained from corn HRs, which was attributed to a more efficient removal of lignin and hemicellulose by a higher sodium hydroxide concentration, which facilitates the dissolution of amorphous cellulose during acid hydrolysis. MCC obtained from HRs exhibited lower bulk density and poorer flow properties but similar or better tableting properties compared to commercial MCC (Ceolus TM PH101). The lower ejection and detachment stress suggests that MCC isolated from HRs requires less lubricant compared to commercial MCC. This study showed that MCC isolated from wheat and corn HRs exhibits comparable tableting behaviour like commercial sample, further supporting this type of agricultural waste utilization.

Leptospirosis is a (re) emerging zoonosis that occurs worldwide. This study aimed to assess seroprevalence of leptospirosis and to identify the most common reactive serovars and risk factors for seropositivity in apparently healthy stray dogs of unknown vaccination status in the Sarajevo region of Bosnia and Herzegovina. Positive microscopic agglutination test titres (≥ 1:25) were detected in 3.87% (156/4028) of samples and most of the sera reacted against one serovar (85.9%). Dogs were most commonly reactive to Canicola (40.4%) and Hardjo (33.3%), followed by Pomona (15.4%) Tarassovi (14.7%), Icterohaemorrhagiae (8.3%), Grippotyphosa (5.8%), Bratislava (1.3%) and Saxkoebing (0.6%). Dogs older than one year had higher odds of seropositivity compared to younger dogs. The seropositivity was higher in spring and autumn than in summer. These results advocate for the need of a control strategy for this zoonosis in the country, which should include sero-surveillance, monitoring, and the inclusion of additional serovars in the testing., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study employed the MTT assay to assess the cytotoxicity of one flavan and two stilbene derivatives isolated from the false indigo-bush ( Amorpha fruticosa L.) fruits: 5,7-dihydroxy-8-geranylflavanone ( 1 ), 2-carboxy-3,5-dihydroxy-4-geranylbibenzyl ( 2 ), and 2-carboxy-3-hydroxy-4-prenyl-5-methoxybibenzyl ( 3 ). The examined compounds reduced the survival of human cervical and colon tumour cells (HeLa, HT-29, HCT-116, and LS174) with IC 50 values ranging from 10.55 to 147.09 μg/mL, except for 1 , which did not affect LS174 cells within the tested concentrations. The highest activity was observed for 1 against HeLa cells, and 1 also exhibited the weakest effect against normal foetal lung fibroblasts (IC 50 = 166.11 μg/mL), demonstrating good potency and selectivity. Stilbenes 2 and 3 proved efficacious, but lacked selectivity compared to 1 . Our findings revealed the cytotoxicity of false indigo-bush constituents, justifying further mechanistic and in vivo investigations, particularly on 5,7-dihydroxy-8-geranylflavanone, which displayed considerable in vitro anticancer capacity and a potentially favourable safety profile.

The present study aimed to evaluate the antioxidant capacity of ethanolic extracts from post-harvest sunflower and maize stalk residues, and their impact on the chemical composition, physicochemical parameters, lipid oxidative stability, microbiological properties, and sensory characteristics of pork liver pâtés over a 90-day storage period. Four formulations were prepared: a control group (CON), a batch with butylated hydroxytoluene as a synthetic antioxidant (BHT), 1% ethanolic extract from sunflower residues (SSRE), and 1% ethanolic extract from maize residues (MSRE). The MSRE had a higher total phenol content and showed better antioxidant activity relative to the SSRE ( p < 0.01). The addition of SSRE decreased the lightness and increased the redness in the pork liver pâtés, with these pâtés showing the highest total color difference compared to the control ( p < 0.01). The crop extracts increased the n-6 and total PUFA contents in pâtés and improved the PUFA/SFA ratio ( p < 0.01). Formulations containing crop residue extracts showed higher TBARs and POV values than the control and BHT group ( p < 0.01), indicating a pro-oxidant effect and accelerated lipid oxidation in pâtés during storage. As far as microbiological quality, the presence of crop residue extracts decreased the total viable count, lactic acid bacteria, and psychotropic aerobic bacteria ( p < 0.01). The incorporation of crop extracts in the pork pâtés impaired their sensory quality, particularly color, odor, aroma, and flavor, and decreased their overall acceptability. These results indicated that, while the crop residue extracts were not as effective as synthetic antioxidants in preserving the lipid stability of pâtés, they demonstrated potential for enhancing the microbial quality of this type of meat product.

To obtain improved insights into the complex microbial aetiology of bovine mastitis, this study investigated the pathogens involved in cattle mastitis in Bosnia and Herzegovina. A total of 179 milk samples from cows with clinical mastitis (CM) and subclinical mastitis (SCM), as well as eight bulk tank milk (BTM) samples from 48 dairy farms, were analysed by standard bacteriological and mycological methods. Mycoplasma detection and identification were performed using culture techniques and real-time polymerase chain reaction (PCR). A total of 88 (49.2%) mastitis samples were positive for known mastitis pathogens at 32 of 47 farms (68.1%). Mycoplasma bovis was a predominant pathogen (25/187; 13.4%) in the majority of herds (14/48; 29.2%) and accounted for 48.9% of positive CM samples. Escherichia coli was the second most dominant CM pathogen (34%), followed by Streptococcus agalactiae (10.6%), whereas Staphylococcus aureus and coagulase-negative staphylococci were the most common in SCM samples (17.1%). Other mastitis pathogens included Candida spp. and Prototheca zopfii . Two BTM samples were positive for M. bovis only, and one was positive for a mixed culture of S. aureus and Streptococcus uberis . The finding of various causative agents of bovine mastitis, with M. bovis emerging as the main pathogen, emphasizes the significance of comprehensive testing that includes not only common mastitis pathogens but also mycoplasmas, fungi, and algae.

Waxes, phospholipids, free fatty acids, peroxides, aldehydes, soap, trace metals and moisture present in crude sunflower oil have a negative effect on the oil quality and are, therefore, removed during the refining process. Waxes crystallizing at low temperatures are removed during winterization by cooling and filtration. Waxes have poor filtration characteristics and an industrial filtration process must be enhanced by the use of filtration aids, which improve filter cake structure and properties, and consequently prolong the filtration cycle. Today, traditional filtration aids (diatomite, perlite, etc.) being used in the industry are frequently replaced by cellulose-based aids. The aim of this study is to examine the effect of oil filtration assisted by two cellulose-based filtration aids on the chemical parameters (wax, moisture, phospholipids, soaps, and fatty acids), oil transparency, carotenoids, and Fe and Cu content of sunflower oil obtained in an industrial horizontal pressure leaf filter. In order to investigate the mentioned parameters, the following techniques were used: gravimetric (waxes and moisture content), spectrophotometric (phospholipids and carotenoid content and oil transparency), volumetric (soaps and free fatty acids content) as well as inductively coupled plasma mass spectrometry (ICP-MS) for Fe and Cu content. An artificial neural network model (ANN) was employed for the prediction of removal efficiency based on the chemical quality, oil transparency, Fe and Cu content in oils before filtration, as well as filtration aid quantity and filtration time. Cellulose-based filtration aids had multiple beneficial effects; on average, 99.20% of waxes, 74.88% of phospholipids, 100% of soap, 7.99% of carotenoids, 16.39% of Fe and 18.33% of Cu were removed.

Mycoplasma ovipneumoniae is an important pathogen in sheep, goats, and wild ruminants. We sequenced M. ovipneumoniae strains 150 and 274 from Bosnia and Herzegovina. Strain 150 has a circular genome of 1,053,380 bp with 29.15% GC content while strain 274 has 1,081,404 bp with 28.82% GC content.

A comparative in vitro study of the antioxidant potential of natural phenols (zingerone, curcumin, raspberry ketone, magnolol) and their synthesized derivatives was performed. The antioxidant efficiency was evaluated in blood serum obtained from healthy individuals, by means of spectrophotometry, before and after the addition of pro-oxidant tert -butyl hydroperoxide (TBH). Moreover, the antioxidant effect of an equimolar mixture of curcumin and zingerone was investigated. Interpretation of our results reveals that in the blood serum of healthy individuals curcumin (C1), raspberry ketone (RK1), magnolol (M1) and synthesized derivative of zingerone (Z2) demonstrate remarkable antioxidant effects ( p < 0.05). However, in the state of TBH-induced excessive oxidative stress natural magnolol and synthesized derivatives C1, Z1 and RK1 show powerful antioxidant activity and thus can be further investigated to obtain information about their metabolic transformations and their potential influence at the cellular level. Results obtained from measurements in an equimolar mixture of zingerone and curcumin indicate synergism ( p < 0.05) between the two compounds. This combination is especially successful due to the fast and efficient neutralization of added pro-oxidant TBH. The commercial availability of turmeric and ginger and their frequent combined use in diet suggest ideas for further broader utilization of the beneficial synergistic effect of their phenolic components.

Mycoplasma ovipneumoniae , a well-established respiratory pathogen of sheep and goats, has gained increased importance recently because of its detection in wild ruminants including members of the Cervidae family. Despite its frequent isolation from apparently healthy animals, it is responsible for outbreaks of severe respiratory disease which are often linked to infections with multiple heterologous strains. Furthermore, M. ovipneumoniae is characterized by an unusually wide host range, a high degree of phenotypic, biochemical, and genomic heterogeneity, and variable and limited growth in mycoplasma media. A number of mechanisms have been proposed for its pathogenicity, including the production of hydrogen peroxide, reactive oxygen species production, and toxins. It shows wide metabolic activity in vitro, being able to utilize substrates such as glucose, pyruvate, and isopropanol; these patterns can be used to differentiate strains. Treatment of infections in the field is complicated by large variations in the susceptibility of strains to antimicrobials, with many showing high minimum inhibitory concentrations. The lack of commercially available vaccines is probably due to the high cost of developing vaccines for diseases in small ruminants not presently seen as high priority. Multiple strains found in affected sheep and goats may also hamper the development of effective vaccines. This review summarizes the current knowledge and identifies gaps in research on M. ovipneumoniae , including its epidemiology in sheep and goats, pathology and clinical presentation, infection in wild ruminants, virulence factors, metabolism, comparative genomics, genotypic variability, phenotypic variability, evolutionary mechanisms, isolation and culture, detection and identification, antimicrobial susceptibility, variations in antimicrobial susceptibility profiles, vaccines, and control.

Ethnopharmacological Relevance: Medicinal properties of hyssop have been used in traditional medicine since ancient times, inter alia, in diseases/conditions with an inherent inflammatory process., Aim of the Study: Accordingly, the aim of this study was to investigate the anti-inflammatory properties of hyssop herb preparations (essential oil and methanol extracts) in vivo, in vitro and in silico., Materials and Methods: For in vitro testing of essential oils and extracts of hyssop herb, the cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) enzyme assays were used. In vivo anti-inflammatory potential of the extracts (at doses of 50, 100 and 200 mg/kg) was assessed using the carrageenan-induced rat paw edema test. Molecular docking and dynamics were used for in silico testing of the inhibitory activity of chlorogenic (CA) and rosmarinic (RA) acids, as the dominant compounds in the tested methanol extracts against COX-1 and COX-2 enzymes., Results: Significant inhibitory activity was shown in the COX-2 test regarding extracts (essential oils did not exhibit any significant activity). Namely, all analyzed extracts, at a concentration of 20 μg/mL, showed a percentage of inhibition of COX-2 enzyme (54.04-63.04%), which did not indicate a statistically significant difference from the positive control of celecoxib (61.60%) at a concentration of 8.8 μM. In vivo testing showed that all methanol extracts of hyssop herb, at the highest test dose of 200 mg/kg in the third and fourth hours, after carrageenan administration, exhibited a statistically significant (p < 0.05) inhibitory effect on the increase in rat paw edema in relation to control. This activity is comparable or higher in relation to the reference substance, indomethacin, at a concentration of 8 mg/kg. The preliminary in silico results suggest that investigated compounds (RA and CA) showed better inhibitory activity against COX-1 and COX-2 than standard non-steroidal anti-inflammatory drug (NSAID), ibuprofen, as evident from the free binding energy (ΔG bind in kJ mol -1 ). The binding energies of the docked compounds to COX-1 and -2 were found to be in the range between -47.4 and -49.2 kJ mol -1 . Ibuprofen, as the one NSAID, for the same receptors targets, showed remarkably higher binding energy (ΔG bind  = -31.3 kJ mol -1 to COX-1, and ΔG bind  = -30.9 kJ mol -1 to COX-2)., Conclusion: The results obtained not only support the traditional use of hyssop herb in inflammatory conditions in folk medicine, but also open the door to and the need for further in vivo testing of extracts in order to examine the molecular mechanism of anti-inflammatory activity in living systems and possibly develop a new anti-inflammatory drug or supplement., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Brucellosis is one of the most common zoonotic diseases worldwide that is endemic to Bosnia and Herzegovina (B&H) and remains an emerging public and animal health concern in this country. Diagnostic testing of brucellosis in ruminants in B&H relies exclusively on serological methods. The present study was conducted to determine the presence of Brucella spp. in clinical samples of seropositive ruminants by Real-time PCR assay. Of the 135 samples tested, 37% were positive by Real-time PCR. The frequency of detection of Brucella-DNA in the samples collected from aborted animals (n = 20/20; 100%) was significantly higher (P < 0.00001) when compared to asymptomatic animals (n = 30/115; 26%). Among asymptomatic animals, 31.1% of cattle were positive for Brucella-DNA, followed by sheep (23.4%) and goats (16.6%). The results of this research underline the limitations of the current control policy and indicate the need for additional diagnostic methods required for more effective brucellosis control program., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Stray dogs may be highly exposed to vector-borne pathogens (VBPs), including zoonotic agents, and therefore may pose a high risk of spreading infections to other animals and humans. Among the Anaplasmataceae, Anaplasma phagocytophilum, A. platys and Ehrlichia canis are commonly identified species in dogs in Europe; however, information on the occurrence of these pathogens in canine populations from Bosnia and Herzegovina (B&H) is still lacking. Thus, the aim of this study was to determine the seroprevalence of Anaplasma spp. and Ehrlichia spp. in stray dogs in the Sarajevo region of B&H and to identify A. phagocytophilum, A. platys, E. canis and E. ewingii by molecular techniques. A total of 903 blood samples of stray dogs were screened by SNAP 4Dx Plus Test for the presence of antibodies against A. phagocytophilum/A. platys and E. canis/E. ewingii. Real-time PCR assays were performed for the detection of Anaplasmataceae, A. phagocytophilum, A. platys, E. canis and E. ewingii in seropositive dogs. Antibodies to A. phagocytophilum/A. platys and/or E. canis/E. ewingii were detected in 187 (20.7%) samples. Seroprevalence was highest for A. phagocytophilum/A. platys (184/903, 20.4%). Two dogs had antibodies to E. canis/E. ewingii, while one dog was found to have antibodies to A. phagocytophilum/A. platys and to E. canis/E. ewingii. Forty-eight (25.7%) of the 187 seropositive dogs examined by Real-time PCR were positive for Anaplasmataceae. A. phagocytophilum was detected in 45 (24%) samples, while one sample was positive for A. phagocytophilum and A. platys. Two samples positive for Anaplasmataceae tested negative in the species-specific PCRs. E. canis or E. ewingii could not be detected in any of the Ehrlichia-seropositive dogs. These findings highlight the need for dog health monitoring, improving the health and welfare of stray dog population, and establishment of effective surveillance systems to combat VBDs., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

The objective of this study was to determine and compare the antimicrobial resistance profiles of clinical Staphylococcus pseudintermedius ( n  = 90) and Staphylococcus aureus ( n  = 61) isolates from different animal species in Bosnia and Herzegovina. Among S. pseudintermedius isolates from dogs ( n  = 86), resistance to penicillin (87.2%) was most common followed by amoxicillin (76.7%) and amoxicillin/clavulanic acid (67.4%). A total of 25 isolates were found to be resistant to oxacillin of which 21 were mecA -positive and multidrug resistant (MDR). The most frequent MDR pattern was penicillins-lincosamides-cephalosporins-macrolides-fluoroquinolones. Among the 61 methicillin-susceptible S. pseudintermedius isolates, 18 were MDR (penicillins-lincosamides-macrolides). Most of S. aureus isolates were resistant to penicillin (50.8%), followed by amoxicillin (41%) and amoxicillin/clavulanic acid (31%). Resistance to cefoxitin was detected in only two isolates. All S. aureus isolates were mecA and mecC- negative. MDR was observed in six S. aureus isolates of which five were from cattle (penicillins-lincosamides-fluoroquinolones). Isolates from red foxes ( Vulpes vulpes ) were sensitive to most of the antimicrobials tested. The high number of methicillin-resistant S. pseudintermedius and MDR isolates in dogs exposed in this study underlines the urgent need for establishment of national antimicrobial resistance surveillance program in animals in the country, as well as for the surveillance of veterinary antimicrobial consumption.