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Pathogenic Yersinia strains were isolated between December 1998 and April 1999 from 37 wild animals: rabbit (Lepus europeus), boar (Sus scrofa scrofa), asiatic jackal (Canis aureus), red fox (Vulpes vulpes), mouflon (Ovis musimon), european river otter (Lutra lutra), beech marten (Martes foina), polecat (Mustela putorius) and wild cat (Felis silvestris). It was established that among the wild animals Y. enterocolitica strains of serotype 0:3 predominated, accompanied by Y. pseudotuberculosis strains of serotype 0:3. In one sample from asiatic jackal and one sample from rabbit, Y. enterocolitica serotype 0:8 was isolated. Yersinia enterocolitica and Y. pseudotuberculosis strains were isolated from tonsils and tongues as well as from the viscera – lung, liver, heart, spleen, kidney and lymph nodes, mainly in young animals (1–2 years of age). The results showed that wild animals are a possible natural reservoir for pathogenic Y. enterocolitica and Y. pseudotuberculosis and are included in the epidemiological chain of yersinioses. [ABSTRACT FROM AUTHOR]

Bovine tuberculosis is still a serious problem with major economic impact in many countries. The aim of study was to evaluate the diagnostic capabilities of the classical and some modern, rapid methods for isolation and identification of Mycobacterium bovis. In the period 2015-2018 from 29 outbreaks in 10 different regions of Bulgaria, 1193 lymph nodes from slaughtered cattle were examined by pathoanatomical, bacteriological, PCR and immunochromatographic methods. Of the 283 bacterial isolates, 263 were identified as M. bovis - member of the M. tuberculosis complex. [ABSTRACT FROM AUTHOR]

Background: This study aimed to characterize recent Mycobacterium bovis/M. caprae isolates from Bulgaria by whole-genome sequencing (WGS) to gain a first insight into their molecular diversity, transmission, and position within the global phylogeography of this important zoonotic species., Results: The isolates were obtained from cattle in diverse locations of Bulgaria in 2015-2020 and were identified by microbiological and PCR assays. WGS data were used for phylogenetic analysis that also included M. bovis global dataset. Thirty-seven M. bovis/caprae isolates from Bulgaria were studied and 34 of them were SNP genotyped. The isolates were subdivided into 3 major phylogenetic groups. Type Mbovis-13 (Eu2 complex [western Europe and northern Africa]) included one isolate. Mbovis-37 type included 5 isolates outside of known clonal complexes. The Bulgarian M. caprae isolates formed a sub-group within the Mcaprae-27B cluster which also included 22 M. caprae isolates from Poland, Spain, Germany, and the Republic of Congo. The Bulgarian M. caprae isolates share their latest common ancestors with Spanish isolates. The Mbovis-37 group shares a distant common ancestor (pairwise distance 22-29 SNPs) with an isolate from Poland but was very distant (> 200 SNPs) from the rest of the tree. The Mbovis-13 group shares a common ancestor with two human isolates from Germany. Phylogeographically, both M. bovis clades had limited circulation in northeastern Bulgaria while the majority of the studied isolates (M. caprae) were from central and western provinces. A phylogenetic network-based analysis demonstrated that 11 Bulgarian isolates were separated by 1 to 6 SNPs within four clusters, mostly forming pairs of isolates., Conclusion: The obtained WGS analysis positioned the Bulgarian isolates within the global phylogeography of M. bovis/M. caprae. Hypothetically, the observed phylogenetic diversity may not have resulted from livestock trade routes, but instead may reflect the deeply rooted M. bovis/M. caprae phylogeography of Europe. A high level of genetic divergence between the majority of the studied isolates suggests limited active transmission of bTB in Bulgaria during the survey period. At the same time, a possibility of the endemic presence of circulating bTB strains in the form of the latent persistent disease cannot be ruled out., (© 2022. The Author(s).)

Bovine tuberculosis (bTB) represents a significant economic burden to the agriculture. In spite of decades of the control program, Mycobacterium bovis infection levels in cattle in Bulgaria continued to rise over recent years. In order to gain a better understanding of the M. bovis diversity, we used spoligotyping for strain differentiation and the data were compared to the international databases Mbovis.org and SITVIT2 for shared type and clade assignment. Study sample included 30 M. tuberculosis complex isolates from cattle originating from different regions of Bulgaria. The isolates were subdivided by spoligotyping into 4 spoligotypes: 2 types shared by 20 and 8 isolates and 2 singletons. SITVIT2-defined types SIT645 and SIT647 belonged to the common and classical bovine ecotype M. bovis (9 isolates) while types SIT120 and SIT339 belonged to the M. caprae ecotype (21 isolates). A certain phylogeographic gradient of the spoligotypes and clades at the within-country level was observed: M. caprae was prevalent in the central/southwestern, while classical M. bovis in the northeastern Bulgaria. Whereas all four types have global or European circulation, neither was described in the neighboring Balkan countries. M. caprae isolates identified in this study mostly belong to the Central/Eastern European cluster. In summary, this study provided a first insight into phylogeography of M. bovis in Bulgaria and described, for the first time, M. caprae as an important infectious agent of bTB in this country., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The study objective was to get more information on C. burnetii prevalence in wild birds and ticks feeding on them, and the potentialities of the pathogen dissemination over Europe by both., Materials: Blood, blood sera, feces of wild birds and ticks removed from those birds or from vegetation were studied at two sites in Russia: the Curonian Spit (site KK), and the vicinity of St. Petersburg (site SPb), and at two sites in Bulgaria: the Atanasovsko Lake (site AL), and the vicinity of Sofia (site SR)., Methods: C. burnetii DNA was detected in blood, feces, and ticks by PCR (polymerase chain reaction). All positive results were confirmed by Sanger's sequencing of 16SrRNA gene target fragments. The antibodies to C. burnetii in sera were detected by CFR (complement fixation reaction)., Results: Eleven of 55 bird species captured at KK site hosted Ixodes ricinus. C. burnetii DNA was detected in three I. ricinus nymphs removed from one bird (Erithacus rubecula), and in adult ticks flagged from vegetation: 0.7% I. persulcatus (site SPb), 0.9% I. ricinus (site KK), 1.0% D. reticulatus (AL site). C. burnetii DNA was also detected in 1.4% of bird blood samples at SPb site, and in 0.5% of those at AL site. Antibodies to C. burnetii were found in 8.1% of bird sera (site SPb). C. burnetii DNA was revealed in feces of birds: 0.6% at AL site, and 13.7% at SR site., Conclusions: Both molecular-genetic and immunological methods were applied to confirm the role of birds as a natural reservoir of C. burnetii. The places of wild bird stopover in Russia (Baltic region) and in Bulgaria (Atanasovsko Lake and Sofia region) proved to be natural foci of C. burnetii infection. Migratory birds are likely to act as efficient "vehicles" in dispersal of C. burnetii -infested ixodid ticks., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

The chemical composition of fresh flowers from Allium ursinum (ramsons, bear's garlic, wild garlic) growing in Bulgaria has been studied. Thymidine (1), adenosine (2), astragalin (kaempferol-3-O-beta-D-glucopyranoside (3), kaempferol-3-O-beta-D-glucopyranosyl-7-O-beta-D-glucopyranoside (4), kaempferol-3-O-beta-D-neohesperoside (5), and kaempferol-3-O-beta-D-neohesperoside-7-O-beta-D-glucopyranoside (6) were isolated from the n-butanol extract and identified by different spectroscopic and spectrometric methods. Thymine (7), uridine (8), uracil (9) and 5-chloro-uridine (10) were detected in the same extract by GC-MS. This is the first report of the occurrence of 1, 2, 4, 7 - 10 in the flowers of A. ursinum. GC-MS of the volatile components of fresh flowers and leaves from the same plant revealed a high content of sulfur compounds, some of which are reported for the first time for A. ursinum. The antimicrobial activities of extracts from fresh flowers and leaves of A. ursinum have been tested; some extracts exhibited moderate antifungal properties.