Different Regions Of Bulgaria Research Articles

Composition and antioxidant potential of essential oil of Geranium macrorrhizum L. from different regions of Bulgaria

The common geranium (Geranium macrorrhizum L.) is the economically most important species of the genus Geranium, and is highly valued for its fragrance and medicinal properties. Object of the present study is the essential oil (EO) prepared by hydro-distillation of aerial parts and rhizomes of common geranium from four floristic regions of the country. Its quality was determined by GC-MS analysis. The antioxidant potential of G. macrorrhizum was evaluated by the DPPH method. Forty-two compounds were identified. In the EOs from the aerial parts, the compound in the most quantity was the sesquiterpene germacrone (from 51.41 to 62.58 %). In the EOs from the rhizomes, this compound was only 5.80-8.94%. The most common ingredient of the rhizome EO was another sesquiterpene: cis-β-elemenone in a quantity from 45.20 to 50.64%. Other nine compounds were present only in the rhizome samples, among which the sesquiterpene globulol was found in good quantities: 15.71 – 15.90%. Monoterpenes like α-terpinene and phellandrene, and sesquiterpenes like eudesm-11-en-4α,6α-diol and eudesm-7(11)-en-4-ol acetate were detected only in some EOs from the aerial parts. The radical scavenging potentials of the EOs were from 33.15 to 41.79 µmol TE/100 µg EO, the rhizome samples showing weaker antioxidant potential compared to the aerial parts. Pearson correlation coefficients between the main ingredients of the EOs from the aerial parts and the radical scavenging potential pointed to the strongest impact of germacrene A on thе antioxidant value. The large differences in the results are due to the different agro-ecological conditions under which the plants were grown.

Phytonutrients in black nightshade fruit (Solanum nigrum L.) growing in Bulgaria

Black nightshade (Solanum nigrum L.) belongs to the Solanaceae family and is a medicinal plant with wild populations found in different regions of Bulgaria. The aim of this study was to analyze the availability of selected phytonutrients in the fruits, and, in particular, their presence in two individual fruit structures (seeds and peels) regarded as by-products in juice production. Fresh fruits were characterized by their diameter (8.42±0.62 mm) and absolute weight (393.75±38.00 g/1000 pcs). The seeds contained 9.81% lipids, while their content in the peels was negligible. The main fatty acids in the extracted lipid fraction were linoleic (48.0%), oleic (27.8%) and palmitic (19.6%), with 76.6% share of unsaturated fatty acids. β-Sitosterol (70.80%) predominated in the sterol fraction, and γ-tocopherol (100%) – in the tocopherol fraction of the extracted oil. The main soluble carbohydrates in the seeds were glucose (2.78%) and fructose (2.42%), while the respective quantities in the peels were 4.21% and 3.82%. The cellulose content of the two fruit parts was 14.90% and 15.70%, respectively. The most abundant macro minerals in the seeds were K (15109.34 mg/kg) and Mg (1988.07 mg/kg), and the main micro minerals – Fe (53.68 mg/kg) and Zn (37.77 mg/kg). The peels contained the respective dietary minerals in substantially higher quantities; K (31373.32 mg/kg), Na (3348.46 mg/kg), Ca (2365.82 mg/kg). The results from the study contribute to a more complex appreciation of the nutritional potential of black nightshade fruit available in Bulgaria and the prospects for their processing into value-added products

Damages on hazelnut kernels caused by Penicillium spp.

The presence of Penicillium spp. was observed during a laboratory examination of nuts (Corylus avellana L.) collected from April 2019 to April 2023 from hazelnut trees, grown in different regions of Bulgaria. The obtained isolates were identified following the Koch’s postulates and their cultural and morphological characteristics were studied. Three-hundred ninety seven (397) samples, with suspected presence of pathogen, were taken from nuts. Penicillium spp. strains were identified as a cause for damages in approximately 2% of all examined kernels. Keywords: Penicillium spp., hazelnut, pathogenicity

Regional health and social inequalities in Bulgaria

Abstract Background The administrative division of Bulgaria divides it into 28 regions, which differ significantly in terms of both socioeconomic development and a number of demographic indicators. The death rate is an indicator according to which our country has been at the top of the world rankings for several years. Cardiovascular diseases stand out among the leading causes of death and they are one of the most important problems facing our society and health system. The study aims to analyse the relationship between the death rate due to CVD in different regions in Bulgaria with the average annual salary of employed persons as an indicator for the socioeconomic development of the regions. Methods The regional differences in the death rate due to CVD and the average annual salary of employed persons were analysed based on the National Statistical Institute data. Analysis and calculations were performed by MS Excel 2019. Results The death rate due to CVD in Bulgaria varies widely - from 1739.7 per 100000 in region Vidin, 1685,2 in region Montana to 878.0 in the city of Sofia, with a national average of 1163.2 /2021/. At the same time, Vidin and Montana regions are also among the regions with a very low average annual salary of employed persons - respectively 12 855 and 13 983 BGN, which is much lower than the average for the country - 21 128 and than the capital Sofia - 25 724 BGN /2021/. Conclusions The regional health inequalities in Bulgaria are very important in order to control and decrease the level of mortality in the country, especially the death rate due to CVD. Bulgaria is among the counties that have the most serious challenges related to high cardiovascular deaths and should be among the priorities of public policy. The profound regional differences that were found impose permanent monitoring and directing the regional social and public health policies to avoid their unfavourable effects. Society should be making major and cost-effective efforts to reduce mortality. Key messages • The elimination of regional inequalities in the country is one of the possible ways to deal with the difficult demographic situation which occurred in the years after the democratic changes. • Regional health inequalities create the need for a long-term health policy that will improve the population's health status and the effective functioning of the healthcare system in our country.

The Role of Family and Media Environment on Aggressive Behaviour in Bulgarian Schools

The article examines the influence of two of the main social environments on students’ aggressive behaviours. On the one hand, attention is paid to the general socio-psychological climate in the family environment and the attitude of the parents towards the child; and on the other hand, a less frequently studied aspect related to aggressive manifestations of the children and adolescents towards the parents is addressed. The study explores how watching TV shows and movies, as well as video games, in which aggression and violence predominate, is connected to the frequency and degree of manifestation of types of aggression in adolescents. The survey was conducted at the end of 2017 among 992 students in 18 primary schools, secondary schools, and vocational schools/high schools in six different cities in different regions of Bulgaria. A structured questionnaire for the study of aggression in school was designed and was intended for students. The results of the study generally show that the verbal aggression towards parents (insulting and shouting) is mostly associated with verbal aggression towards both teachers and classmates. Regarding the influence of TV shows and movies containing aggression, it was found that students who watched movies with military, fighting and bloody scenes demonstrated more frequent manifestations of verbal aggression, but the TV contents did not significantly influence the manifestations of indirect aggression and physical aggression. However, the frequency of playing video games with aggressive content has a significant effect on all investigated forms of aggression, with the strongest effect on physical aggression. Aggression in social networks is a significant factor that affects the frequency of manifestation of various forms of aggression. Students who bully others on social networks stand out as the most aggressive (verbally, physically, and indirectly).

Potato Viruses of Economic Importance for Production of Planting Material in Bulgaria

Potato is the world’s third most widespread food crop, following wheat and rice. Tubers play an important role in the spread of virus diseases in different regions in Bulgaria for seed production. The most important viruses in potatoes in Bulgaria include Potato virus Y (PVY), Potato leaf roll virus (PLRV), Potato virus S (PVS), and Potato virus M (PVM). Most of the viruses in potatoes do not induce symptoms in tubers, which plays an important role in viral dissemination by the growers. In this way, after planting the tubers, the quality and yield of the production may be severely reduced. The use of virus-free tubers by growers is of key importance for the control of diseases and the reduction of loss of production.

Comparative Study on Physicochemical, Antioxidant and Antimicrobial Properties of Propolis Collected from Different Regions of Bulgaria

Abstract Propolis (bee glue), a valuable biological mixture produced by European honey bees (Apis mellifera L.), is widely used as a remedy in traditional and alternative medicine, as an effective food biopreservative and as a nutritional value enhancer. The present study aimed to investigate eighty propolis samples collected from all twenty-eight districts of Bulgaria in the period of 2020–2022 in order to determine their physicochemical properties as well as their antioxidant and antimicrobial potential. The raw propolis samples exhibited brown or green colour depending on the climatic zone from which they are collected and pH values from 4.82 to 5.87, while the moisture content ranged between 0.98% and 2.97%. The total phenolics content (TPC) and total flavonoids content (TFC) varied from 63.14 mg GAE/g to 737.27 mg GAE/g, and from 29.22 mg QE/g to 234.17 mg QE/g, respectively. The propolis samples demonstrated significant antioxidant potential determined by DPPH and FRAP methods, from 18.56 mM TE/g to 1598.66 mM TE/g and from 82.28 mM TE/g to 1208.81 mM TE/g, respectively, whose values showed a positive correlation (r 2) with the TPC and TFC. The results from the antimicrobial screening revealed that the methanolic propolis extracts in concentration of 20 mg/ml exhibited the highest inhibitory effect on the Gram-positive bacteria such as Micrococcus luteus 2YC-YT, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 25923, Listeria monocytogenes NBIMCC 8632 and Listeria innocua ATCC 33090.

Detection of HEV RNA Using One-Step Real-Time RT-PCR in Farrow-to-Finish Pig Farms in Bulgaria.

(1) Background: HEV is a zoonotic, foodborne pathogen. It is spread worldwide and represents a public health risk. The aim of this study was to evaluate the presence of HEV RNA in farrow-to-finish pig farms in different regions of Bulgaria; (2) Methods: Isolation of HEV RNA from pooled samples of feces was performed using a QIAamp® Viral RNA Mini Kit followed by HEV RNA detection using a single-step real-time RT-PCR with primers and probes targeting the ORF 3 HEV genome; (3) Results: HEV RNA was detected in 12 out of 32 tested farms in Bulgaria (37.5%). The overall percentage of HEV-positive pooled fecal samples was 10.8% (68 of 630 samples). HEV was detected mostly in pooled fecal samples from finisher pigs (66/320, 20.6%) and sporadically from dry sows (1/62, 1.6%) and gilts (1/248, 0.4%); (4) Conclusions: Our results confirm that HEV circulates in farrow-to-finish pig farms in Bulgaria. In our study, we found HEV RNA in pooled fecal samples from fattening pigs (4-6-months age), shortly before their transport to the slaughterhouse indicating a potential risk to public health. The possible circulation of HEV throughout pork production requires monitoring and containment measures.

Infections with Dicrocoelium dendriticum, Cysticercus pisiformis, Micipsella numidica and seroprevalence of Toxoplasma gondii in European brown hares (Lepus europeus L.) from Bulgaria

During the period 2009–2016, brown hares (Lepus europeus L.) from different regions of Bulgaria were investigated for infections with liver parasites as well as for seroprevalence of protozoan Toxoplasma gondii. Three helminth species were found in the liver: one trematode – Dicrocoelium dendriticum, one cestode – larval form of Taenia pisiformis – Cysticercus pisiformis and one nema­tode – Micipsella numidica. Dicrocoeliasis was found in all parts of the country with infected hares originating mainly from the western regions. The total prevalence of infection with D. dendriticum was 10.59%. The intensity of infection ranged from 1 to 184, 26.67 on the average. The total prevalence of infection with C. pisiformis was 16.8%. The prevalence of infection was highest among hares from the southern parts of the country, and no cysticercosis was registered in the western parts. The extent of infestation with cysticerci was different – from single cysticerci on the omentum or serosa of some organs in the abdominal cavity to innumerable ones that affected also the organs in the thoracic cavity. However, cases in which the animals were slightly affected, predominated. The total prevalence of infection with M. numidica was 5.31%, and the intensity of infection varied between 1 and 6 parasites in one animal. The nematodiasis was established mainly in hares from southern Bulgaria. The total seroprevalence of T. gondii was 6.15% and the protozoan-positive animals originated from three districts: Stara Zagora, Plovdiv, and Burgas. The current studies also showed that the prevalence of D. dendriticum infection among animals increased with age. No correlation was observed between prevalence of infection with C. pisiformis and M. numidica and host age. No correlation was found between the prevalence of the helminth infections and sex of the animals.

Phytochemical analysis of leaves and stems of Physalis alkekengi L. (Solanaceae)

Abstract Physalis alkekengi L. (Solanaceae) is encountered in different regions of Bulgaria as a wild growing or ornamental plant. The objective of this work was to characterize the phytochemical composition (macro and micro components) of the leaves and stems of two local phenotypes (PA-SB and PA-NB), with the view of revealing their use potential. The dry leaves contained (DW) protein (16.25 and 19.27%), cellulose (25.16 and 25.31%), and ash (18.28 and 16.16%) and the stems contained protein (6.83 and 7.35%), cellulose (39.34 and 38.25%), and ash (15.01 and 7.48%) for PA-SB and PA-NB, respectively. The dominant amino acids (by HPLC) in the leaves of both phenotypes were arginine (21.3–22.3 mg/g) and aspartic acid (8.8–18.4 mg/g), and those in the stems were proline and aspartic acid for PA-SB (8.8, 7.7 mg/g); isoleucine and tyrosine for PA-NB (12.8, 6.6 mg/g). Mineral elements, determined by AAS (K, Ca, Mg, Na, Cu, Fe, Zn, Mn, Pb, Cr), also varied between phenotypes and plant parts. The leaves alone were further processed by extraction with n-hexane, for the identification of leaf volatiles (by gas chromatography-mass spectrometry). The analysis identified 28 components (97.99%) in the leaf extract of PA-SB and 32 components (97.50%) in that of PA-NB. The volatile profile of PA-SB leaves was dominated by diterpenes (49.96%) and oxygenated sesquiterpenes (35.61%), while that of PA-NB was dominated by oxygenated aliphatics (40.01%) and diterpenes (35.19%). To the best of our knowledge, the study provides the first data about the phytochemical composition of the leaves and stems of P. alkekengi from Bulgaria, in a direct comparison of phenotypes from two distinct wild populations, which could be of further scientific interest.

Detection of measles virus in Bulgaria from to 2018

AimTo determine the circulation patterns of measles virus in Bulgaria from 2012 to 2018 after a large measles outbreak in the country (2009-2011).MethodsThree types of clinical material were collected: serum samples, urine samples, and nasal swabs. Enzyme-linked immunosorbent assay (ELISA) was used to detect specific viral immunoglobulin (Ig) M/IgG antibodies. Viral RNA was extracted from all urine and nasal swabs.ResultsIn the investigated period, 102 patients were confirmed to have measles (age range: two months to 55 years). A total of 101 samples (99%) were measles-IgM positive. Most of them were detected in 2017 (73%, 74/101), when a measles outbreak in the country was reported. The majority of patients were unvaccinated children aged under 13 months. Out of 101 measles serum samples confirmed by ELISA, 18 (20.45%) were measles-IgG positive and 15 (17.05%) were borderline. Thirty-three positive PCR products were sequenced and genotyped. In 2013, 2016, 2017, and 2018, three different measles viral genotypes were detected: D8, H1, and B3. Most patients were unvaccinated or insufficiently vaccinated.ConclusionPreventive measures are indispensable to limit the infection in different regions of Bulgaria and its spread to other countries. As vaccination coverage against measles and other vaccine-preventable infections, including SARS-Co2, is low, it is necessary to perform molecular identification of viruses to monitor their circulation and pathogenicity.

Prevalence of Salmonella spp. in Breeding Flocks, Laying Hens and Broilers in Different Regions of Bulgaria in 2016

In 2016 a total number of 297 samples were tested in the frame of Salmonella control programmes in Bulgaria including official control (n = 101) and control by the business operators (n = 196). The samples originated from 3 categories of hen flocks: 48 pcs. samples from breeding flocks, 175 pcs. from laying hens and 74 pcs. from broilers. The samples belonged to 3 different groups: I – fresh feces, boot swabs and fabric socks swabs (n = 281), II – dust samples (n = 10), and III – egg shells (n = 6). The laboratory tests were performed in accordance with EN ISO 6579: 2003/À1:2007. Salmonella positives were found in 40% of dust samples (n = 10) and 2.09% of fresh feces, boot swabs, fabric socks swabs, and egg shells (n = 287). The percentage of Salmonella spp. positive samples was 3.37%, including 0.67% of breeding flocks and 2.69% of laying hens. The prevalence of Salmonella in breeding flocks was 4.17% and among laying hen herds – 4.57%. There was no Salmonella spp. detected in broilers. The ratio of isolated Salmonella serovars was: Enteritidis – 60%, Senftenberg – 20%, and Corvallis and Infantis – 10%. In total 6.93% of the samples tested for official controls (n = 101) and 1.53% for business operator control (n = 196) were positive for Salmonella. It is recommended the replacement of dust samples with another type of sample to be solely as an exception and the control of Salmonella spp. in samples from the primary production to be as much as possible under the official control.

Exploring the Potential of Straw Biochar for Environmentally Friendly Fertilizers

The pyrolysis of wheat straw in order to produce biochar for soil amendment is a potential strategy for producing environmental friendly fertilizers capable of boosting soil fertility, increasing carbon storage, and lowering greenhouse gas emissions. However, straw biochar’s potential to influence these aspects may vary depending on its properties. Our study sought to investigate biochar from wheat straw from three different regions in Bulgaria. A specially designed set up was used for the biochar production. Three pyrolytic temperatures (300, 400, and 500 °C) were applied, resulting in nine biochar samples. The specific characteristics included moisture content, volatile substances content, ash content, fixed carbon content, and joint ash and carbon content, and they were determined for each sample. The chemical content, resulting in 17 chemical elements and compounds, was measured and analysed. The results obtained showed that the produced straw biochar has the potential to be used as a fertilizer and soil supplement.

Antioxidant Activity and Chemical Characteristics of Sambucus nigra L. Blossom from Different Regions in Bulgaria

The aim of the current study was to investigate some bioactive compounds from Sambucus nigra L. blossoms and to evaluate the antioxidant potential of the obtained extracts. In this study, samples from four different regions of Bulgaria /Rhodopes, Plovdiv, Strandzha and Dobrich region/ from Sambucus nigra L. were collected and analyzed for total phenols, flavonoids, sugars and amino acids. The antioxidant activity of the extracts was evaluated by four assays based on different mechanisms. The sweetness index and total sweetness index of the extracts were also evaluated. The carbohydrate composition of the leaves and the blossoms was determined, with glucose and fructose predominating in both cases, as their contents were not above 3%. Nineteen amino acids have been identified in the composition of Sambucus nigra L., and glutamic, leucine and asparagine acids are predominant. The highest antioxidant activity and total content of phenols (49.2 ± 1 mg GAE/g) and flavonols (18.6 ± 0.5 mgQE/g) were found in the sample from the Rhodope region. Therefore, the higher altitude and lower temperature in mountains could influence the accumulation of secondary metabolites in blossoms of Sambucus nigra L., which improves the antioxidant potential of the samples.

Development of the cherry leaf spot epidemics in different regions of Bulgaria

A 2-year research on the epidemics of the cherry leaf spot (CLS) was conducted in unsprayed cherry orchards at two locations in Bulgaria (Krichim and Gabarevo). The primary infections for both years at Gabarevo happened in mid-May during fruit development. In Krichim during 2018 the first symptoms of the disease appeared in early June (fruit ripening), while in 2019 they became evident in the second half of May (fruit development). In both locations and years, the period of a rapid increase in the CLS incidence and defoliation was from mid-June till late July. The maximum of the disease incidence and defoliation was observed in early September. More seasonal rather than regional trend in the disease dynamics was observed. In 2018 the orchards in both locations exhibited delayed the CLS progression compared to 2019 due to the relatively dry conditions in April and May. The higher disease incidence and defoliation in 2019 could be assigned to the high infection background and weather conditions in the beginning of the growing season.

STREPTOCOCCUS SPP. AS ETIOLOGICAL AGENT OF SUBCLINICAL AND CLINICAL MASTITIS OF DAIRY COWS IN REPUBLIC OF BULGARIA

The aim of this study was to determine the species composition of streptococci causing subclinical, and clinical mastitis in dairy cows, their prevalence and antimicrobial drug sensitivity with respect to appropriate therapy. The study subject was the involvement and prevalence of bacteria from the genus Streptococcus in the etiology of subclinical, and clinical mastitis in dairy cows. The survey was conducted in eight cattle farms from different regions in Bulgaria in 2020 and 2021. After clinical examination and rapid mastitis test, a total of 239 cows were diagnosed with udder inflammation - 34 (14.2%) with clinical signs and 205 (85.8%) with subclinical mastitis. Laboratory analyses were performed on 346 samples. Two hundred and seventy two bacterial strains were isolated, and 151 (55.5%) were determined to belong to Streptococcus spp. Lancefield grouping and species identification of haemolytic variants was done. It was found out that in subclinical forms of mastitis, 54.5% of isolates belonged to Streptococcus spp., respectively 63.3 % of isolates in clinical forms of mastitis also belonged to Streptococcus spp. Out of them, 38.6% were S. uberis, 35.6% were identified as S. agalactiae and 25.8% − as S. dysgalactiae. In vitro tests of the sensitivity to antimicrobial drugs from various groups and classes were conducted with all field isolates. The tests demonstrated a relatively well-preserved sensitivity to antimicrobial agents with the exception of tetracycline

Bacillus Strains as an Effective Treatment of Mobile Forms of Phosphorus in Bulgarian Soils

Strains of genus Bacillus have the potential to increase the availability of phosphorus to plants by the dissolution of inorganic phosphate, which favours the growth of plant species and that has an important economic and agricultural impact. Soils with low phosphate intensity (Lеаched chernozem) were enriched with poorly soluble phosphorus compounds and inoculated with the tested newly isolated strains from genus Bacillus (Bacillus subtilis T 2, Bacillus amyloliquefaciens T 3, Bacillus subtilis T 4, Bacillus subtilis T 10, Bacillus thuringiensis T 17 and Bacillus cereus Т 18). Six newly isolated strains from different regions in Bulgaria were identified by classical phenotypic techniques and 16S rDNA sequence analysis. Tested strains were inoculated (2 ml and 15 ml) in the soil and incubated for 25 days at 28oC. After the incubation period, the degradation of phosphorite flour to available phosphorus was examined by the classical method of Egner-Riehm and by extraction with CaCl2. A vegetation experiment was conducted with a test plant Pelargonium zonale, characterized by its ability to absorb large amounts of phosphorus. A peat substrate enriched with all macro-and microelements was used, and phosphorus was added to the medium in the form of phosphorite flour. The amounts of phosphorus absorbed by one plant Pelargonium zonale treated with newly isolated strains Bacillus subtilis T 10 and Bacillus cereus T 18 absorbed 23% more phosphorus than the control variant. As a result of the experiments, it was found that the studied strains have a positive impact on the increase of phosphorus mobility in soils with low phosphate intensity treated with hardly degradable phosphors.

Molecular Detection of Nosema spp. in Honey in Bulgaria

Environmental DNA (eDNA) analysis is related to screening genetic material of various organisms in environmental samples. Honey represents a natural source of exogenous DNA, which allows for the detection of different honey bee pathogens and parasites. In the present study, we extracted DNA from 20 honey samples from different regions in Bulgaria and tested for the presence of DNA of the ectoparasitic mite Varroa destructor, as well as Nosema apis and Nosema ceranae. Only Nosema ceranae was detected, showing up in 30% of all samples, which confirms the widespread prevalence of this pathogen. All positive samples were found in plain regions of the country, while this pathogen was not detected in mountainous parts. None of the samples gave positive amplifications for the Nosema apis and Varroa mite. The obtained results from this study confirm previous observations that eDNA contained in honey is a potent source for effective biomonitoring of actual diseases in the honey bee.

Radioactivity of Drinking Waters from Different Regions of Bulgaria in the Period 2017-2019

Radioactivity of Drinking Waters from Different Regions of Bulgaria in the Period 2017-2019

Molecular Epidemiology of Xanthomonas euvesicatoria Strains from the Balkan Peninsula Revealed by a New Multiple-Locus Variable-Number Tandem-Repeat Analysis Scheme.

Bacterial spot of pepper and tomato is caused by at least three species of Xanthomonas, among them two pathovars of Xanthomonas euvesicatoria, which are responsible for significant yield losses on all continents. In order to trace back the spread of bacterial spot pathogens within and among countries, we developed the first multilocus variable number of tandem repeat analyses (MLVA) scheme for pepper- and tomato-pathogenic strains of X. euvesicatoria. In this work, we assessed the repeat numbers by DNA sequencing of 16 tandem repeat loci and applied this new tool to analyse a representative set of 88 X. euvesicatoria pepper strains from Bulgaria and North Macedonia. The MLVA-16 scheme resulted in a Hunter–Gaston Discriminatory Index (HGDI) score of 0.944 and allowed to resolve 36 MLVA haplotypes (MTs), thus demonstrating its suitability for high-resolution molecular typing. Strains from the different regions of Bulgaria and North Macedonia were found to be widespread in genetically distant clonal complexes or singletons. Sequence types of the variable number of tandem repeats (VNTR) amplicons revealed cases of size homoplasy and suggested the coexistence of different populations and different introduction events. The large geographical distribution of MTs and the existence of epidemiologically closely related strains in different regions and countries suggest long dispersal of strains on pepper in this area.