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This study investigated the phytochemical characteristics, antibacterial activity, and synergistic potential of essential oils derived from Romanian lavender. Gas Chromatography-Mass Spectrometry (GC/MS) analysis revealed that linalool is the main compound in all lavender essential oils, with concentrations ranging from 29.410% to 35.769%. Linalyl acetate was found in similar concentrations to linalool. Other significant compounds included 1,8-cineole (8.50%), lavandulyl acetate (5.38%), trans-ß-ocimene (6.90%), and camphor (7.7%). A 1,1-Diphenyl-2-Picrylhydrazyl (DPPH) test was used to assess antioxidant capacity, with substantial free-radical-scavenging activity shown in the IC50 values determined. The antibacterial efficacy of the oils was higher against Gram-positive bacteria than Gram-negative bacteria, with variations in minimum inhibitory concentrations (MICs), the extent of inhibition, and evolution patterns. The study also explored the oils' ability to enhance the efficacy of ampicillin, revealing synergistic interactions expressed as fractional inhibitory concentration indices. In silico protein-ligand docking studies used twenty-one compounds identified by GC-MS with bacterial protein targets, showing notable binding interactions with SasG (-6.3 kcal/mol to -4.6 kcal/mol) and KAS III (-6.2 kcal/mol to -4.9 kcal/mol). Overall, the results indicate that Romanian lavender essential oils possess potent antioxidant and antibacterial properties, and their synergistic interaction with ampicillin has potential for enhancing antibiotic therapies.

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Bee products are considered true wonders of nature, used since ancient times, and studied even today for their various biological activities. In this study, we hypothesise that Romanian bee products from different origins (micro apiary products, lyophilised forms, commercial) exhibit distinct chemical compositions, influencing their biological activities. An LC-MS analysis revealed varied polyphenolic content patterns, with cumaric acid, ferulic acid, rosmarinic acid, and quercitine identified in significant amounts across all samples. Primary anti-inflammatory evaluation phases, including the inhibition of haemolysis values and protein denaturation, unveiled a range of protective effects on red blood cells (RBC) and blood proteins, contingent upon the sample concentration. Antimicrobial activity assessments against 12 ATCC strains and 6 pathogenic isolates demonstrated varying efficacy, with propolis samples showing low efficacy, royal jelly forms displaying moderate effectiveness, and apilarnin forms exhibiting good inhibitory activity, mostly against Gram-positive bacteria. Notably, the lyophilised form emerged as the most promising sample, yielding the best results across the biological activities assessed. Furthermore, molecular docking was employed to elucidate the inhibitory potential of compounds identified from these bee products by targeting putative bacterial and fungal proteins. Results from the docking analysis showed rosmarinic and rutin exhibited strong binding energies and interactions with the putative antimicrobial proteins of bacteria (-9.7 kcal/mol to -7.6 kcal/mol) and fungi (-9.5 kcal/mol to -8.1 kcal/mol). The findings in this study support the use of bee products for antimicrobial purposes in a biologically active and eco-friendly proportion while providing valuable insights into their mechanism of action.

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The research aimed to determine the chemical composition, the antioxidant and anti-inflammatory activity as well as the antimicrobial activity against Gram-positive, Gram-negative and two fungal Candida ATCC strains of a commercial Boswellia essential oil (BEO) containing Boswellia carteri, Boswellia sacra, Boswellia papryfera, and Boswellia frereana. Additionally, molecular docking was carried out to show the molecular dynamics of the compounds identified from the essential oil against three bacterial protein targets and one fungal protein target. The major components identified by GC-MS (Gas Chromatography-Mass Spectrometry) were represented by α-pinene, followed by limonene. Evaluation of antioxidant activity using the DPPH (2,2-Diphenyl-1-Picrylhydrazyl) method showed high inhibition comparable to the synthetic antioxidant used as a control. Oxidative stability evaluation showed that BEO has the potential to inhibit primary and secondary oxidation products with almost the same efficacy as butylated hydroxyanisole (BHA). The use of BEO at a concentration of 500 ppm provided the best protection against secondary oxidation during 30 days of storage at room temperature, which was also evident in the peroxide value. Regarding the in vitro anti-inflammatory activity, the membrane lysis assay and the protein denaturation test revealed that even if the value of protection was lower than the value registered in the case of dexamethasone, the recommendation of using BEO as a protective agent stands, considering the lower side effects. Gram-positive bacteria proved more sensitive, while Pseudomonas aeruginosa presented different sensitivity, with higher MICs (minimal inhibitory concentration). Haemophilus influenzae demonstrated a MIC at 2% but with consecutive inhibitory values in a negative correlation with the increase in concentration, in contrast to E. coli, which demonstrated low inhibitory rates at high concentrations of BEO. The computational tools employed revealed interesting binding energies with compounds having low abundance. The interaction of these compounds and the proteins (tyrosyl-tRNA synthetase, DNA gyrase, peptide deformylase, 1,3-ß-glucan synthase) predicts hydrogen bonds with amino acid residues, which are reported in the active sites of the proteins. Even so, compounds with low abundance in BEO could render the desired bioactive properties to the overall function of the oil sustained by physical factors such as storage and temperature. Interestingly, the findings from this study demonstrated the antioxidant and antimicrobial potential of Boswellia essential oil against food-related pathogens, thus making the oil a good candidate for usage in food, feed or food-safety-related products.

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Canine morbillivirus, also known as canine distemper virus (CDV), is the causative agent of canine distemper (CD), which is a serious contagious disease of canines, large felids, and, occasionally, raccoons. This study included seven raccoons from the Timisoara Zoological Garden, Romania. CDV was detected using RT-qPCR on blood samples, but several other exams were also performed-clinical, bacteriological, immunohistochemistry (IHC) and histopathology, toxicological screening, and necropsy-which confirmed CDV infection. Severe digestive disorders (diarrhea and frequent hematemesis) were observed. The necropsy findings included pseudo membranous gastroenteritis, congestion, and pulmonary edema in two raccoons. Immunohistochemistry showed immunolabeled CDV antigenantibodies on the viral nucleocapsid. Histopathology revealed lymphocyte depletion in mesenteric lymphnodes and intranuclear and intracytoplasmic inclusions in the enterocytes of the small intestine. Based on the RT-qPCR assay, laboratory tests, and the lesions observed, it was established that the raccoons were infected with CDV, which was the cause of death in two cases. The results from the necropsy, histology, and immunohistochemistry in the raccoons are comparable with reported CDV lesions in dogs. In conclusion, several exams may be performed to establish the etiology of possible interspecific viral infection, but only very specific exams can identify aCDV infection. Laboratory analyses must be completed by RT-qPCR assay or IHC to establish infection with uncommon viruses in raccoons with high accuracy.

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Honey is a natural product with multiple health benefits. The paper presents the chemical characterization and the antioxidant and antimicrobial potential of ten types of honey (knotweed, linden, wild cherry, acacia, honeydew, oilseed rape, sunflower, phacelia, plain polyflora and hill polyflora) from the Banat region, Romania. We studied the water content, dry matter, impurities, acidity and pH of honey. We also determined the content of reducing sugar, minerals and flavonoids and the total phenolic content. All honey samples analysed showed good nutritional characteristics according to the standard codex for honey. From the analysis of the mineral content of the honey samples, we observed a variability in the macro and microminerals, influenced by the botanical origin, ranging between 0.25% (wild cherry honey) and 0.54% (honeydew). The toxic metals' (Cd and Pb) levels met the standard for almost all samples analysed except for knotweed. The flavonoid content of the samples ranged from 9.29 mg QE/100 g for wild cherry honey to 263.86 mg QE/100 g for linden honey, and for polyphenols between 177.6 mgGAE/100 g for acacia honey and 1159.3 mgGAE/100 g for honeydew. The best antioxidant capacity was registered in the case of linden honey (79.89%) and honeydew (79.20%) and the weakest in acacia (41.88%) and wild cherries (50.4%). All studied honey samples showed antimicrobial activity, depending on the type of honey, concentration and strain analysed. The novelty of this study is given by the complex approach of the study of honey quality, both from the perspective of chemical attributes and the evaluation of the antimicrobial potential on specific strains in correlation with the botanical and geographical origin of the analyzed area.

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Staphylococcal infection treatment in dogs is frequently associated with adverse side effects, high costs, prolonged treatment, and resistant strain selection. Staphylococcus aureus is the most frequently isolated staphylococci in cases of canine superficial pyoderma. The number of Staphylococcus strains to exhibit primary resistance to various drugs in vitro is increasing. Propolis has a diverse chemical composition and well-known therapeutic properties against bacterial infections. The current investigation evaluated in vitro the antimicrobial activity of the commercial allopathic antimicrobials, Romanian propolis ethanolic extracts, against clinical Staphylococcus aureus strains isolated from superficial dermatitis clinical samples in dogs and two reference strains: Staphylococcus aureus ATCC 25923 and Staphylococcus aureus ATCC 43300, as the MSSA and MRSA positive controls, respectively, in western Romania. We used the microdilution broth technique to evaluate the susceptibility profile of the bacteria. The minimum inhibitory concentration (MIC) of the Romanian propolis ethanolic extract ranged from 6 to 10 µg/mL for all isolates, determined by the broth microdilution method. The MICs of ethanolic Romanian propolis extracts had a pronounced antibacterial activity. These results indicate that propolis can potentially be used and recommended for in vivo experiments as a promising therapeutic agent against Staphylococcus aureus infections in superficial dermatitis of dogs.

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This study aimed to investigate the antioxidant profile and the antimicrobial activity of four different types of monofloral honey (manuka (MH), brassica rapeseed (BH), acacia (AH), and linden honey (LH)) against some bacterial/fungal ATCC strains and some multidrug-resistant strains isolated from chronic otitis in dogs. For the characterisation of the antioxidant profile of each honey, we extracted the honey samples by hydroalcoholic extraction and analysed them in terms of total polyphenols (TPC), total flavonoids (TFC), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) using the spectrophotometric method. The antimicrobial activity was determined using the microdilution method at concentrations of 10%, 15%, and 20%, with the results expressed in OD (optical density) calculated as BIR% (bacterial inhibition rate)/MIR% (mycelial inhibition rate). The antioxidant characterisation of the analysed honey samples showed the highest antioxidant activity and concentrations of TPC and TFC in MH, followed by LH. MH was proven to be the most effective on most clinical isolates concerning the antimicrobial activity in comparison with BH, AH, and LH. Except for B. cepacia and P. vulgaris, all the clinical isolates were sensitive to the antibacterial activity of honey. Regarding the ATCC strains, MH 10% was the most effective in inhibiting all the strains tested except for P. aeruginosa. In conclusion, the efficacy classification in our study was MH > BH > AH > LH.

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This study aimed to investigate the chemical composition and the activity against Staphylococcus aureus (S. aureus) (ATCC 25923), Streptococcus pyogenes (S. pyogenes) (ATCC 19615), Escherichia coli (E. coli) (ATCC 25922), Pseudomonas aeruginosa (P. aeruginosa) (ATCC 27853), Shigella flexneri (S. flexneri) (ATCC 12022), Salmonella typhimurium (S. typhimurium) (ATCC 14028), Haemophillus influenzae (H. influenza) type B (ATCC 10211) and two fungal strains: Candida albicans (C. albicans) (ATCC 10231) and Candida parapsilopsis (C. parapsilopsis) (ATCC 22019) of the extracts obtained from Melilotus officinalis (MO), Coronilla varia (CV); Ononis spinosa (OS) and Robinia pseudoacacia (RP) (Fabaceae), and to identify the chemical compounds responsible for the antimicrobial effect against the tested strains. The extracts were obtained by conventional hydroalcoholic extraction and analyzed in terms of total polyphenols using the spectrophotometric method and by liquid chromatography (LC). The results have shown that the highest polyphenols content was recorded in the RP sample (16.21 mg gallic acid equivalent GAE/g), followed by the CV (15.06 mg GAE/g), the OS (13.17 mg GAE/g), the lowest value being recorded for the MO sample (11.94 mg GAE/g). The antimicrobial testing of plant extracts was carried out using the microdilution method. The most sensitive strains identified were: E. coli, S. typhimurium, P. aeruginosa and S. pyogenes, while protocatechuic acid, gallic acid, caffeic acid, quercetin, rutin, and kaempferol were identified as the chemical compounds responsible for the antibacterial effect. The analysis of the correlation between the chemical composition and the antimicrobial effect proved a moderate (r > 0.5) positive correlation between rosmarinic acid and S. pyogenes (r = 0.526), rosmarinic acid and S. typhimurium (r = 0.568), quercetin and C. albicans (r = 0.553), quercetin and S. pyogenes (r = 0.605). Therefore, it suggested possible antimicrobial activity generated by these chemical components. The results recommend the Fabaceae plants as promising candidates for further research to develop novel natural antimicrobial drugs.

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Bacterial contamination of boar semen occurs with some frequency in artificial insemination centers and may have a negative effect on the quality of the semen as well as on the sows' reproductive capacity. Normally, the source of bacterial contamination in pig seminal doses is the own boar. However, distilled water or laboratory equipment used to elaborate the seminal doses can be an important source of bacterial contamination. This study focused on the identification of gram-negative bacteria in boar semen, and impact on the quality of ejaculates obtained from boar, as well as on the establishment of antimicrobial resistance patterns of isolated gram-negative bacteria. Semen samples were collected from 96 boars, ranging in age from 12-36 month, from three artificial insemination centers from the North-West of Romania. Bacterial species were identified by two methods: matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and API 20 E (BioMerieux, France). The main bacteria isolated from the doses diluted semen were gram-negative bacteria (47.91%), with a majority of the contaminant bacteria belonging to the family Enterobacteriaceae: Seratia marcescens 19.56%, Proteus mirabilis 15.21% and Escherichia coli 10.86% and to the family Pseudomonaceae: Ralstonia picketii 17.39%, Burkholderia cepacia 10.86%, Pseudomonas aeruginosa 8.69%, and Pseudomonas fluorescens 4.34%, respectively. More than half of these isolates (56.52%) were resistant to gentamycin and 58.69% were resistant to penicillin. These antibiotics are very frequently added in sperm diluent in the centers for the processing of sperm from boars in Romania. Regular monitoring for bacterial contamination is an important aspect of a control program.