Search

Human umbilical cord (UC) is an attractive source of mesenchymal stromal/stem cells (MSCs) for tissue engineering and regenerative medicine due to its easy availability, non-invasive procedure of collection, and no ethical concerns. The aim of this study was to isolate MSCs from the Wharton’s jelly (WJ) and subamnion (SA) from the same umbilical cord by an optimized explant method, and to compare the morphology, proliferation, and stemness properties of the MSCs from both sources. Cells from the WJ and SA of six umbilical cords were characterized by flow cytometry, differentiation capacity and proliferation assays, immunofluorescence staining, and RT-PCR. The optimized explant method was successfully used to isolate WJ-MSCs and SA-MSCs. The MSCs from both sources showed similar patterns of growth kinetics, adipogenic and osteogenic potential, and the expression of pluripotency markers (OCT4, SOX2, NANOG, and SSEA-4). The current findings support the usage of the optimized explant method to generate a relatively homogenous population of MSCs from Wharton’s jelly and subamnion, which can facilitate the reproducibility of the results from experimental and practical applications of the obtained cells.

The aim of this study was to analyze the seroprevalence of West Nile virus (WNV) among equids in Bulgaria, confirm the results of a competitive ELISA versus the virus neutralization test (VNT) and investigate some predisposing factors for WNV seropositivity. A total of 378 serum samples from 15 provinces in northern and southern Bulgaria were tested. The samples originated from 314 horses and 64 donkeys, 135 males and 243 females, aged from 1 to 30 years. IgG and IgM antibodies against WNV protein E were detected by ELISA. ELISA-positive samples were additionally tested via VNT for WNV and Usutu virus. Thirty-five samples were WNV-positive by ELISA (9.26% [CI = 6.45–12.88]), of which 15 were confirmed by VNT; hence, the seroprevalence was 3.97% (CI = 2.22–6.55). No virus-neutralizing antibodies to Usutu virus were detected among the 35 WNV-ELISA-positive equids in Bulgaria. When compared with VNT, ELISA showed 100.0% sensitivity and 94.5% specificity. A statistical analysis showed that the risk factors associated with WNV seropositivity were the region (p < 0.0001), altitude of the locality (p < 0.0001), type of housing (p < 0.0001) and breed (p = 0.0365). The results of the study demonstrate, albeit indirectly, that WNV circulates among equids in northern and southern Bulgaria, indicating that they could be suitable sentinel animals for predicting human cases and determining the risk in these areas or regions of the country.

The aim of this study was to compare the diagnostic capabilities of the agar gel immunodiff usion test (AGID) and two types of PCR, nested PCR (with nucleic acid extraction from blood) and direct blood nested PCR (db nested PCR) - without extraction, vs. enzyme-linked immunosorbent assay (ELISA) as the gold standard in the routine diagnostics of this disease. A total of 409 blood samples were obtained from cattle 18 mo. to 5 yrs. of age, and all the samples were analyzed using the four assays. Following the initial testing, the samples were stored at -20°С and re-tested using all the four techniques after a month of freeze storage to determine the reproducibility of the results. ELISA detected 57 animals as positive (13.9%) versus 33 ones using AGID and 56 using the two types of nested PCR. AGID showed low sensitivity of 57.9% and moderate agreement compared to ELISA. In addition, AGID did not show consistency in the results from the two independent measurements. The two types of nested PCR showed nearly full agreement with ELISA with a kappa value of 0.99. Since AGID showed lower sensitivity and lack of reproducibility in the results for 22 samples as compared to the other techniques used in this study, we suggest that the future application of this test for the diagnosis of Enzootic bovine leucosis in blood samples should be reconsidered. On the other hand, db nested PCR demonstrated very good sensitivity and reproducibility of results, it also requires less sample processing. All this makes it potentially suitable for routine diagnostics.

The study tracks the utilization of Ocimum basilicum L. (sweet basil)—a garden plant popular for its ritual and ornamental value in the past, that is currently applied in various forms and ways as medicine, food, insect repellent, etc.—in Bulgaria. Previous data for Bulgarian rural home gardens showed a significant number of preserved local landraces; however, it remained unclear how people perceive the large varietal diversity of this species and how the traditions related to its use are preserved. We combined a literature review on the cultural value of sweet basil and the breeding of local genetic resources with an online questionnaire, directed to adult laypeople, that sought to access different aspects of past (recalled) and present use and related knowledge. The identification skills of the participants were tested using images of local plant landraces and foreign varieties. Responses from 220 participants showed that potted “Genovese”-type individual was most frequently identified as sweet basil (89.9%), followed by two examples of local landraces in flower. Participants who grow sweet basil or used it in more varied ways had significantly better identification skills. Ocimum basilicum was most frequently reported as food, while ritual/symbolic use was preserved while devalued during the Communism regime (1945–1989). Food and religious uses were negatively associated in the past, but presently, the tendency is completely reversed. Preferences for the informal exchange of seeds and seed-saving practices were discussed.

The global climate and societal challenges in the recent years urge us to strengthen food security; thus, the rediscovery of wild foods and foraging practices is also part of the sustainability agenda. Utilization of underappreciated sources such as ruderal plants could be a valuable option, especially for vulnerable parts of the society. We present data on traditional knowledge on spring edible ruderal plant taxa preserved in rural regions of Bulgaria, combining field studies in the period 2017–2022 that were compared to the available recent and historical ethnographic and (ethno)botanical literature. Semi-structured interviews were performed with representatives of 94 households in North and South Bulgaria, focusing on collection practices, used parts, and preparation methods. We list 65 edible ruderals, belonging to 22 plant families, of which 19 appeared only in the literature sources. Unlike in the Mediterranean tradition, edible ruderal plants in Bulgaria were regarded unfavorably, as poverty food. Amaranthaceae and Asteraceae were the most represented families, with 10 taxa each. About half of the taxa were collected for their leaves or whole young herbage that is used as pastry fillings, in stewed, and in cooked dishes. Taxa used in raw salads were mostly from the literature sources. The most diverse utilization was recorded in the southern-most regions of Bulgaria, where immediate tasting of the gathered plants was reported by the participants as the way to collect food plants. The bitter ones or those with an unappealing smell were considered non-edible and were avoided. References about biologically active compounds and potential benefits were collected, classified, and discussed in regard to their potential benefits for human health.

Lamiaceae comprises widely distributed medicinal and aromatic plants, many of which are traditionally used in European countries. The current study aimed to document Lamiaceae taxa used in rural Bulgaria (Southeast Europe) and to explore the related local knowledge and cultural practices that influence their utilization for various purposes. Field work included inventory of Lamiaceae diversity in home gardens and semi-structured interviews focused on the cultivation, collection, and utilization practices common among elderly inhabitants of 34 settlements in rural Bulgaria. We report the utilization of 27 Lamiaceae taxa, 9 of which were collected from the wild. Traditional and contemporary ways of utilizing Lamiaceae taxa as culinary and medicinal plants, in herbal teas, as repellents, ritual plants, etc., are presented. Recent knowledge on medicinal properties contributed to the introduction of new taxa in gardens (wild and cultivated), while traditional culinary practices were found to sustain the diversity of local forms (landraces).

Artisan food production, with its unique flavors, is a source of knowledge about sustainable use of natural resources. This is because it reflects the skills of local communities in utilizing these resources (e.g., wild and cultivated plants and autochthonous breeds) under specific environmental conditions for a long period of time. Therefore, the use of local ingredients and the reduction in food miles make traditional food a safer, healthier and more ecofriendly choice for consumers. In the present research, we examined the herbal ingredients in Bulgarian dry-cured meats and discuss their contribution to the flavor and durability of the products. A combination of field data, collected through semi-structured interviews in local communities, and an analysis of the available literature was used to reveal the typicity of 24 artisanal/homemade meat products available mostly in their place of origin. We compared the obtained data to 16 industrial products branded as “traditional”, with 6 of them registered under the European Union quality schemes. The recorded ingredients of plant origin (dry and fresh) belonged to 16 taxa. Most diverse spice mixtures were used in products made of pork meat and in products originating from the southern, and hence warmer, regions of the country. The herbs and spices were combined freely except for Alliums, which were not mixed, and only one species was used per product. Most of the spices used in the artisanal dry meats were sourced from home gardens (some specifically cultivated for that purpose). Those collected from the wild, e.g., Origanum vulgare subp. hirtum (Link) Ietsw., were gathered sustainably in small quantities. The number of spices used in industrial dry meat products was limited to two–three, and was provided by cultivated sources, without exploiting natural populations. Manufacturing of all artisanal products was seasonal to avoid the cold winter weather, a measure which was necessary for the natural air-drying of the meat. The long-lasting effects of the abolishment of artisanal production under Communism, the adaptation of traditional products for industry, and the current challenges and perspectives surrounding artisanal production of meat products were discussed.

The home garden is a unique human-nature interspace that accommodates a diverse spectrum of plant species and provides multiple services to households. One of the most important roles of home gardens is to shelter the agricultural plant diversity that provides for diverse and healthy nutrition, especially in rural communities. While tropical home gardens have received wide recognition due to their provisional function for the local communities, temperate and especially European home gardens have been discussed less frequently as a source of subsistence. The main objectives of the current study were to document plant species grown in Bulgarian rural home gardens and to explore related local knowledge and cultural practices that influence food plant diversity, its selection and preservation. Field work was focused on settlements situated in eight provinces in South and North-West Bulgaria. Participants representing 65 home gardens were approached through semi-structured interviews. Home gardens were found to harbor 145 cultivated and semi-cultivated plant taxa, used as food, medicinal and aromatic plants and as animal fodder. Members of the Rosaceae family were most numerous. The largest part of the garden area was occupied by vegetable crops of Solanaceae and Cucurbitaceae. In 63.1% of the studied households, the food growing area comprised more than 2/3 of the total size of the garden. Most preferred crops reflected the social and cultural importance of food self-provisioning, especially in the rural areas. The provisional role of the home gardens in regard to preparation of traditional foods and the driving forces for seed saving are discussed.

Oregonin is an open-chain diarylheptanoid isolated from Alnus incana bark that possesses remarkable antioxidant and anti-inflammatory properties, inhibits adipogenesis, and can be used in the prevention of obesity and related metabolic disorders. Here, we aimed to investigate the effects of oregonin on the epigenetic regulation in cells as well as its ability to modulate DNA methylating enzymes expression and mitochondrial DNA (mtDNA) copies. Our results show that oregonin altered the expression of DNA methyltransferases and mtDNA copy numbers in dependency on concentration and specificity of cells genotype. A close correlation between mtDNA copy numbers and mRNA expression of the mtDnmt1 and Dnmt3b was established. Moreover, molecular modeling suggested that oregonin fits the catalytic site of DNMT1 and partially overlaps with binding of the cofactor. These findings further extend the knowledge on oregonin, and elucidate for the first time its potential to affect the key players of the DNA methylation process, namely DNMTs transcripts and mtDNA.

The addition of prostaglandin F2α (PGF2α) to boar semen prior to insemination improves the conception and farrowing rates in sows. It is accepted that this is due to increased myometrial contractility, which improves the spermatozoa movement. However, there are limited data about the effect of the exogenous PGF2α analogs on sperm motility parameters and morphology. The aim of the current study was to define if there are changes in motility, morphology and kinematic parameters of spermatozoa on 1stand 24th hour after addition of PGF2α analogue to extended boar semen. A total of 18 ejaculates, obtained from clinically healthy boars were diluted 1:3 in semen extender, and each of them was separate into four aliquots, 50 ml each. PGF2α was added to 3 of them in concentrations of 6, 12 and 25 μg/ml, and the fourth served as untreated control. The motility, kinematic parameters and morphology of spermatozoa were evaluated on 1stand 24th hours after addition of PGF2α. There was no significant difference in sperm morphology, total and progressive motility between the untreated and treated groups. There was however a significant decrease in the rapid velocity and some of the kinematic parameters (VCL, VSL and VAP) in the group treated with 25 μg/ml compared to the control at the 1st hour after PGF2α treatment, which (except for the rapid velocity) persisted to the 24th hour. The results indicate that addition of Oestrophan (Bioveta,CZ) to the extended boar semen did not improve the sperm motility, morphology and kinematic parameters of the spermatozoa.