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SARS-CoV-2 emerged in 2019 and found diagnostic laboratories unprepared worldwide. To meet the need for timely and accurate virus detection, laboratories used rapid Ag tests and PCR kits based on costly multi-channel real-time techniques. This study aimed to develop a conventional nested PCR based on the SARS-CoV-2 N gene, validate it against some approved assays, and apply it to samples from six cats with respiratory symptoms obtained in early 2020 during the first COVID-19 wave in humans in Bulgaria. The nested PCR technique showed 100% sensitivity and specificity; it could detect extracted SARS-CoV-2 RNA at concentrations as low as 0.015 ng/μL. The results identified the six tested cat samples as positive. Sequence analysis performed in two of them confirmed this. The presented technique is reliable, easy to implement and inexpensive, and can be successful in strategies for the prevention and control of SARS-CoV-2 in humans, cats and other susceptible species.

Objectives: A total of 226 carbapenem-resistant Acinetobacter baumannii (CRAB) isolates was collected during 2014-2016 from inpatients (age range 5-88 years) in four Bulgarian university hospitals (H1-H4) to assess their antimicrobial susceptibility and to explore carbapenem resistance mechanisms as well as the molecular epidemiology of the isolates., Methods: Antimicrobial susceptibility testing, multiplex PCR, DNA sequencing and electrotransformation experiments were performed. Epidemiological typing by random amplification of polymorphic DNA (RAPD)-PCR was also performed., Results: The resistance rates were as follows: imipenem, 90.7%; meropenem, 98.2%; doripenem, 100%; amikacin, 92.9%; gentamicin, 87.2%; tobramycin, 55.8%; levofloxacin, 98.2%; trimethoprim/sulfamethoxazole, 86.3%; tigecycline, 22.1%; colistin, 0%; and ampicillin/sulbactam, 41.6%. Intrinsic bla OXA-51-like genes were found in all of the isolates. The majority of the A. baumannii isolates harboured either bla OXA-23-like associated with the upstream-located ISAba1 (26.1%) or bla OXA-40/24-like (46.7%), 45 isolates (19.9%) harboured both genes, and 1 isolate harboured bla OXA-58-like surrounded by ISAba3C upstream and ISAba3 downstream. The bla OXA-58 gene was transferable by electroporation indicating its plasmid location. Epidemiological typing revealed the dissemination of nosocomial CRAB with high clonal relatedness (70% similarity threshold) belonging to six, four, three and two clusters in H1, H2, H3, and H4 hospitals, respectively., Conclusions: The A. baumannii isolates studied were problematic nosocomial pathogens. Their multidrug resistance greatly limits therapeutic options. The persistence of endemic clones comprised of OXA carbapenemase-producing multidrug-resistant A. baumannii in the monitored hospitals over a period of ca. 3 years is of concern and requires continuous detailed investigations in the future., (Copyright © 2018 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.)

More than 20 years after the first outbreaks, the phylogenetic picture of PRRSV is still incomplete and full of gaps, especially in regards of PRRSV 1. Due to the exceptional diversity observed at the eastern borders of Europe and the low number of available sequences from Central Eastern European countries, the authors collected and analyzed both recent as well as already submitted sequences comparing them to a large backbone set of available ORF5 sequences representing the full spectrum of PRRSV 1 Subtype 1 diversity to conduct a systematic phylogenetic analysis and reclassification elucidating the diversity of the virus in these countries. Moreover, further analyses of the EUROSTAT data regarding the live pig movement trends revealed their influence of virus diversity and evolution. The results indicate that besides the effect of local, isolated divergent evolution and the use of modified live vaccines, the most important factor influencing a given country's virus diversity is the transboundary movement of live, infected animals.

Captive snakes, that is, a Jamaican boa (Epicrates subflavus) a yellow anaconda (Eunectes notaeus) and a corn snake (Pantherophis guttatus guttatus), died with signs of bacteraemia including the presence of petechial haemorrhages in the mouth and gums and haemorrhages in the lung, spleen and intestines. The abdomen and anus were swollen with bloody-tinged mucus in the colon. Aeromonas hydrophila was recovered in dense virtually pure culture growth from the internal organs. Characterization of the isolates was by phenotyping and sequencing of the 16S rRNA gene (sequence homology of 99% with A. hydrophila) with outputs confirming the identity as A. hydrophila. Pathogenicity experiments confirmed virulence to frogs (Rana esculenta) and rainbow trout (Oncorhynchus mykiss)., (© 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.)

The aim of this study is to investigate the profile of ovine PrP gene by amino acid polymorphism at codons 136, 141, 154, and 171 for determining the genetic predisposition to the Scrapie disease for the tribal sheep and rams, with different numbers and distribution in Bulgaria. Three hundred twenty four animals originating from 41 tribal herds comprising eight breeds were included in the study. DNA was isolated from blood samples specifically amplified by PCR and sequenced. The alignments of codons 136, 141, 154, and 171 were determined. Based on the sequencing, it was established that Bulgarian breeds belong to the second and third risk groups, those with low and moderate risk of Scrapie disease. Establishment of 11 genotypes in Synthetic Population Bulgarian Milk breed reveals it to have the highest risk of the Scrapie disease; moreover, the conducting of the program will be more difficult in comparison with other investigated breeds. Evidence for the internal cross breeding is the presence of the five or six genotypes in the Copper-Red Shoumen, Replian, Karakachan, and Duben Bulgarian native breeds.