The principal aim of this study was to analyze the chemical composition and qualitative and quantitative variability of essential oils obtained from seven naturally grown populations of the Pinus peuce Grisebach, Pinaceae in Kosovo. Plant materials were collected from three populations in the Sharri National Park and from four other populations in the Bjeshkët e Nemuna National Park, in Kosovo. Essential oils were obtained by steam distillation and analyzed by GC-FID (Gas Chromatography-Flame Ionization Detection) and GC-MS (Gas Chromatography-Mass Spectrometry). The results showed that the yield of essential oils (v/w dry weight) varied depending on the origin of population and the plant organs and ranged from 0.7 to 3.3%. In total, 51 compounds were identified. The main compounds were α-pinene (needles: 21.6-34.9%; twigs: 11.0-24%), β-phellandrene (needles: 4.1-27.7; twigs: 29.0-49.8%), and β-pinene (needles: 10.0-16.1; twigs: 6.9-20.7%). HCA (Hierarchical Cluster Analysis) and PCA (Principal Component Analyses) were used to assess geographical variations in essential oil composition. Statistical analysis showed that the analyzed populations are grouped in three main clusters which seem to reflect microclimatic conditions on the chemical composition of the essential oils.

Widespread antibiotic resistance is on the rise and current therapies are becoming increasingly limited in both scope and efficacy. Methicillin-resistant Staphylococcus aureus (MRSA) represents a major contributor to this trend. Quorum sensing controlled virulence factors include secreted toxins responsible for extensive damage to host tissues and evasion of the immune system response; they are major contributors to morbidity and mortality. Investigation of botanical folk medicines for wounds and infections led us to study Schinus terebinthifolia (Brazilian Peppertree) as a potential source of virulence inhibitors. Here, we report the inhibitory activity of a flavone rich extract "430D-F5" against all S. aureus accessory gene regulator (agr) alleles in the absence of growth inhibition. Evidence for this activity is supported by its agr-quenching activity (IC50 2-32 μg mL(-1)) in transcriptional reporters, direct protein outputs (α-hemolysin and δ-toxin), and an in vivo skin challenge model. Importantly, 430D-F5 was well tolerated by human keratinocytes in cell culture and mouse skin in vivo; it also demonstrated significant reduction in dermonecrosis following skin challenge with a virulent strain of MRSA. This study provides an explanation for the anti-infective activity of peppertree remedies and yields insight into the potential utility of non-biocide virulence inhibitors in treating skin infections.

Methicillin-resistant Staphylococcus aureus (MRSA) presents one of the most serious health concerns worldwide. The WHO labeled it as a "high-priority" pathogen in 2017, also citing the more recently emerged vancomycin-intermediate and -resistant strains. With the spread of antibiotic resistance due in large part to the selective pressure exerted by conventional antibiotics, the use of antivirulence strategies has been recurrently proposed as a promising therapeutic approach. In MRSA, virulence is chiefly controlled by quorum sensing (QS); inhibitors of QS are called quorum quenchers (QQ). In S. aureus, the majority of QS components are coded for by the accessory gene regulator (Agr) system. Although much work has been done to develop QQs against MRSA, only a few studies have progressed to in vivo models. Those studies include both prophylactic and curative models of infection as well as combination treatments with antibiotic. For most, high efficacy is seen at attenuating MRSA virulence and pathogenicity, with some studies showing effects such as synergy with antibiotics and antibiotic resensitization. This minireview aims to summarize and derive conclusions from the literature on the in vivo efficacy of QQ agents in MRSA infection models. In vitro data are also summarized to provide sufficient background on the hits discussed. On the whole, the reported in vivo effects of the reviewed QQs against MRSA represent positive progress at this early stage in drug development. Follow-up studies that thoroughly examine in vitro and in vivo activity are needed to propel the field forward and set the stage for lead optimization.

A shortage of conventional medicine during the American Civil War (1861–1865) spurred Confederate physicians to use preparations of native plants as medicines. In 1863, botanist Francis Porcher compiled a book of medicinal plants native to the southern United States, including plants used in Native American traditional medicine. In this study, we consulted Porcher’s book and collected samples from three species that were indicated for the formulation of antiseptics: Liriodendron tulipifera, Aralia spinosa, and Quercus alba. Extracts of these species were tested for the ability to inhibit growth in three species of multidrug-resistant pathogenic bacteria associated with wound infections: Staphylococcus aureus, Klebsiella pneumoniae, and Acinetobacter baumannii. Extracts were also tested for biofilm and quorum sensing inhibition against S. aureus. Q. alba extracts inhibited growth in all three species of bacteria (IC50 64, 32, and 32 µg/mL, respectively), and inhibited biofilm formation (IC50 1 µg/mL) in S. aureus. L. tulipifera extracts inhibited biofilm formation (IC50 32 µg/mL) in S. aureus. A. spinosa extracts inhibited biofilm formation (IC50 2 µg/mL) and quorum sensing (IC50 8 µg/mL) in S. aureus. These results support that this selection of plants exhibited some antiseptic properties in the prevention and management of wound infections during the conflict.

Streptococcus pneumoniae (pneumococcus) forms organized biofilms to persist in the human nasopharynx. This persistence allows the pneumococcus to produce severe diseases such as pneumonia, otitis media, bacteremia and meningitis that kill nearly a million children every year. While bacteremia and meningitis are mediated by planktonic pneumococci, biofilm structures are present during pneumonia and otitis media. The global emergence of S. pneumoniae strains resistant to most commonly prescribed antibiotics warrants further discovery of alternative therapeutics. The present study assessed the antimicrobial potential of a plant extract, 220D-F2, rich in ellagic acid, and ellagic acid derivatives, against S. pneumoniae planktonic cells and biofilm structures. Our studies first demonstrate that, when inoculated together with planktonic cultures, 220D-F2 inhibited the formation of pneumococcal biofilms in a dose-dependent manner. As measured by bacterial counts and a LIVE/DEAD bacterial viability assay, 100 and 200 µg/ml of 220D-F2 had significant bactericidal activity against pneumococcal planktonic cultures as early as 3 h post-inoculation. Quantitative MIC’s, whether quantified by qPCR or dilution and plating, showed that 80 µg/ml of 220D-F2 completely eradicated overnight cultures of planktonic pneumococci, including antibiotic resistant strains. When preformed pneumococcal biofilms were challenged with 220D-F2, it significantly reduced the population of biofilms 3 h post-inoculation. Minimum biofilm inhibitory concentration (MBIC)50 was obtained incubating biofilms with 100 µg/ml of 220D-F2 for 3 h and 6 h of incubation. 220D-F2 also significantly reduced the population of pneumococcal biofilms formed on human pharyngeal cells. Our results demonstrate potential therapeutic applications of 220D-F2 to both kill planktonic pneumococcal cells and disrupt pneumococcal biofilms.

Background Ethnoveterinary knowledge is highly significant for persistence of traditional community-based approaches to veterinary care. This is of particular importance in the context of developing and emerging countries, where animal health (that of livestock, especially) is crucial to local economies and food security. The current survey documents the traditional veterinary uses of medicinal plants in the Lesser Himalayas-Pakistan. Methods Data were collected through interviews, focus groups, participant observation, and by administering questionnaires. A total of 105 informants aged between 20–75 years old who were familiar with livestock health issues (i.e. farmers, shepherds, housewives and herbalists) participated in the study. Results A total of 89 botanical taxa, belonging to 46 families, were reported to have ethnoveterinary applications. The most quoted families were Poaceae (6 taxa), Fabaceae (6), Asteraceae (5), and Polygonaceae (5). Adhatoda vasica was the most cited species (43%), followed by Trachyspermum ammi (37%), and Zanthoxylum armatum var. armatum (36%). About 126 medications were recorded against more than 50 veterinary conditions grouped into seven categories. The highest cultural index values were recorded for Trachyspermum ammi, Curcuma longa, Melia azedarach, Zanthoxylum armatum var. armatum and Adhatoda vasica. The highest informant consensus factor was found for pathologies related to respiratory and reproductive disorders. Comparison with the local plant-based remedies used in human folk medicine revealed that many of remedies were used in similar ways in local human phytotherapy. Comparison with other field surveys conducted in surrounding areas demonstrated that approximately one-half of the recorded plants uses are novel to the ethnoveterinary literature of the Himalayas. Conclusion The current survey shows a remarkable resilience of ethnoveterinary botanical knowledge in the study area. Most of the species reported for ethnoveterinary applications are wild and under threat. Thus, not only is it imperative to conserve traditional local knowledge of folk veterinary therapies for bio-cultural conservation motives, but also to assist with in-situ and ex-situ environmental conservation initiatives, which are urgently needed. Future studies that focus on the validation of efficacy of these ethnoveterinary remedies can help to substantiate emic concepts regarding the management of animal health care and for rural development programs. Keywords: Medicinal plants; Ethnobotany; Ethnoveterinary; Lesser Himalayas; Pakistan

Background: Plant material from different organs of Pistacia terebinthus L., (Anacardiaceae) were collected in Kosovo with aim to analyze the chemical variability of the essential oils among native populations and to test them for potential antibacterial activity against Staphylococcus aureus. Methods: Essential oils obtained from leaves, pedicels, fruits and galls were analyzed by GC-FID and GC/MS. Minimum inhibitory concentration (MIC) against three clinically relevant strains of S. aureus (NRS385, LAC and UAMS-1) were used to evaluate the antibacterial activity of essential oils. Results: In total, 33 different compounds were identified. The main constituents were α-pinene (12.58-66.29 %), D-limonene (13.95-46.29 %), β-ocimene (0.03-40.49 %), β-pinene (2.63-20.47 %), sabinene (0.00-5.61 %) and (Z)-β-ocimene (0.00-44.85 %). Antibacterial testing of the essential oils against three clinical isolates of S. aureus revealed that seven of the eight samples had some activity at the concentration range tested (0.04-0.512 % v/v). The gall tissues from both sites produced the highest yield of essential oil (3.24 and 6 %), and both exhibited growth inhibitory activity against S. aureus. The most bioactive essential oils, which exhibited MIC90 values ranging from 0.032-0.128 % v/v, obtained from the fruits of the Ura e Shejtë collection site. Likewise, the leaf and pedicel essential oil from the same site was highly active with MIC90 values of 0.064-0.128 and 0.032-0.256 % v/v, respectively. Conclusions: Principle Component Analyses demonstrated that there is a variation in the chemical composition of essential oil depending on the plant organs from which essential oil are obtained and the geographical origin of the plant populations. The highest variability regarding the chemical composition of essential oil was found between oils obtained from different organs originating from the Prizren site. The MIC90 activity of Pistacia terebinthus was on par or superior compared with Tea Tree Oil control (0.128 % v/v), suggesting that essential oils from this species may have some potential for development as an antibacterial agent for S. aureus infections.

Background: Kosovo represents a unique hotspot of biological and cultural diversity in Europe, which allows for interesting cross-cultural ethnobotanical studies. The aims of this study were twofold: 1) to document the state of traditional knowledge related to local (esp. wild) plant uses for food, medicine, and handicrafts in south Kosovo; and 2) to examine how communities of different ethnic groups in the region (Albanians, Bosniaks/Gorani, and Turks) relate to and value wild botanical taxa in their ecosystem. Methods: Field research was conducted in 10 villages belonging to the Prizren municipality and 4 villages belonging to the Dragash municipality, located in the Sharr Mountains in the southern part of Kosovo. Snowball sampling techniques were used to recruit 139 elderly informants (61 Albanians, 32 Bosniaks/Gorani and 46 Turks), for participation in semi-structured interviews regarding the use of the local flora for medicinal, food, and handicraft purposes. Results: Overall, we recorded the local uses of 114 species were used for medicinal purposes, 29 for food (wild food plants), and 20 in handicraft activities. The most important species used for medicinal purposes were Achillea millefolium L., Sambucus nigra L., Urtica dioica L., Tilia platyphyllos Scop. Hypericum perforatum L., Chamomilla recutita (L.) Rauschert, Thymus serpyllum L. and Vaccinium myrtillus L. Chamomilla recutita was the most highly valued of these species across the populations surveyed. Out of 114 taxa used for medicinal purposes, only 44 species are also included in the European Pharmacopoeia. The predominantly quoted botanical families were Rosaceae, Asteraceae, and Lamiaceae. Comparison of the data recorded among the Albanian, Bosniak/Gorani, and Turkish communities indicated a less herbophilic attitude of the Albanian populations, while most quoted taxa were quoted by all three communities, thus suggesting a hybrid character of the Kosovar plant knowledge. Conclusion: Cross-cultural ethnobiological studies are crucial in the Balkans not only for proposing ways of using plant natural resources, which could be exploited in sustainable local development projects (e.g. focusing on eco-tourism and small-scale trade of medicinal herbs, food niche and handicrafts products), but also for fostering collaboration and reconciliation among diverse ethnic and religious communities.

Background: Ethnoveterinary medicine is a topic of growing interest among ethnobiologists, and is integral to the agricultural practices of many ethnic groups across the globe. The ethnoveterinary pharmacopoeia is often composed of ingredients available in the local environment, and may include plants, animals and minerals, or combinations thereof, for use in treating various ailments in reared animals. The aim of this study was to survey the current day ethnoveterinary practices of ethnic Hungarian (Székely) settlements situated in the Erdovidék commune (Covasna County, Transylvania, Romania) and to compare them with earlier works on this topic in Romania and other European countries. Methods: Data concerning ethnoveterinary practices were collected through semi-structured interviews and direct observation in 12 villages from 2010 to 2014. The cited plant species were collected, identified, dried and deposited in a herbarium. The use of other materials (e.g. animals, minerals and other substances) were also documented. Data were compared to earlier reports of ethnoveterinary knowledge in Transylvania and other European countries using various databases. Results: In total, 26 wild and cultivated plants, 2 animals, and 17 other substances were documented to treat 11 ailments of cattle, horses, pigs, and sheep. The majority of applications were for the treatment of mastitis and skin ailments, while only a few data were reported for the treatment of cataracts, post-partum ailments and parasites. The traditional uses of Armoracia rusticana, Rumex spp., powdered sugar and glass were reported in each village. The use of some plant taxa, such as Allium sativum, Aristolochia clematitis, and Euphorbia amygdaloides was similar to earlier reports from other Transylvanian regions. Conclusions: Although permanent veterinary and medical services are available in some of the villages, elderly people preferred the use of wild and cultivated plants, animals and other materials in ethnoveterinary medicine. Some traditional ethnoveterinary practices are no longer in use, but rather persist only in the memories of the eldest subset of the population. A decline in the vertical transmission of ethnoveterinary knowledge was evident and loss of practice is likely compounded by market availability of ready-made pharmaceuticals.