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Introduction: Bulgaria joined European Union (EU) on 1 January 2007. Since the accession all regulations, directives, decisions, recommendations, and opinions of the European Parliament, the Council of the European Union, and the European Commission are being implemented.

Background/Aim. Mediterranean spotted fever (MSF) belongs to Rickettsioses, the Spotted fever group (SFG). The causal agent is Rickettsia conorii conorii and the transmission to humans occurs through dog tick Rhipicephalus sanguineus bites. The aim of this study was to describe clinical and laboratory characteristics in patients with severe form of Mediterranean spotted fever admitted to Bulgarian university hospital in endemic region. Methods. A retrospective study was conducted at Stara Zagora University Hospital (Southeastern Bulgaria) between April 2015 and August 2016. During the analyzed period, 58 cases had clinical and laboratory data for MSF. Serological tests were applied for the etiological diagnosis. MSF-specific immunoglobulin (IgM) and IgG antibodies were detected in serum by indirect immunoenzyme assay (ELISA IgG/IgM, Vircell, Spain) – R. conorii ELISA IgG sensitivity 85%, specificity 100% and R. conorii ELISA IgM sensitivity 94%, specificity 95%. Statistical analysis was made by MS Excel 2007 and SPSS Statistics, version 19.0. Results. Eighteen patients presented as severe forms. The predominant gender of them were males (78%) and 22% were females. The median age of the analyzed group was 55 years (range: 14– 78 years). Ten patients developed hepatic disorder while 4 had neurological signs. Laboratory data showed thrombocytopenia in 15 patients, mean value of platelet (PLT) count for the whole group was 108.6 ± 53.8 × 109/L. Liver enzymes were elevated with mean value of aspartate aminotransferase (AST) 161.4 ± 90.1 IU/L and alanine aminotransferase (ALT) 163.9 ± 81.5 IU/L. Acute phase reactant as C-reactive protein (CRP) had mean value of 140.3 mg/L (range: 9–230 mg/L). Kidney function was impaired in some cases; the mean value of creatinine for the studied group was 134.7 μmol/L (range: 78–313 μmol/L) and mean value of urea was 9.6 mmol/L (range: 4.2–27.4 mmol/L). Conclusion. Bulgaria is an endemic area for tick-borne diseases. Cases of MSF are reported annually. Severe forms of MSF are not rare. Typical clinical and laboratory markers for severity should be actively searched for. Early diagnosis and proper treatment is the key to avoid complications and enable patient recovery.

The aim of this study was to assess the prevalence of Coxiella burnetii infection among veterinary medicine students from two Bulgarian Universities, located in Sofia and Stara Zagora. Blood samples were collected from a total of 185 veterinary students for the detection of C. burnetii phase II antibodies and presence of DNA using an enzyme-linked immunosorbent assay (ELISA) and end-point PCR test. Out of all samples, 29.7% were positive for at least one C. burnetii phase II antibody marker or by the result of the PCR test. Veterinary students from Stara Zagora showed a significantly high seropositivity for Q fever (33.6%), as compared to the students in Sofia (23%; p < 0.05). Evidence of recent exposure with detection of anti-C. burnetii phase II IgM (+) antibodies was observed in 14.6% of the students under study. Seroprevalence among students in Stara Zagora was higher (15.3%). Anti-C. burnetii phase II IgG antibodies were detected in 21.6% of examined samples. Our study revealed a higher seropositivity among the male students (32.8%) as compared to females (16.0%; p < 0.05). The end-point PCR assay detected 5.9% blood samples as positive. The relative risk (RR) of Q fever exposure for male students was 40.7%, whereas it was 24.6% in females (p < 0.05). The findings from this study indicate that the C. burnetii infection is widely distributed amongst veterinary students in Bulgaria. This study emphasizes the need for improved safety protocols and infection control measures in veterinary training programs. [ABSTRACT FROM AUTHOR]

In recent years, hepatitis E virus (HEV) infection has been found to be widespread among different animal species worldwide. In Bulgaria, high HEV seropositivity was found among pigs (60.3%), wild boars (40.8%), and East Balkan swine (82.5%). The aim of the present study was to establish the seroprevalence of HEV among dogs, cats, horses, cattle, sheep, and goats in Bulgaria. In total, 720 serum samples from six animal species were randomly collected: dogs—90 samples; cats—90; horses—180; cattle—180; sheep—90; and goats—90. The serum samples were collected from seven districts of the country: Burgas, Kardzhali, Pazardzhik, Plovdiv, Sliven, Smolyan, and Stara Zagora. The animal serum samples were tested for HEV antibodies using the commercial Wantai HEV-Ab ELISA kit (Beijing, China). The overall HEV seroprevalence among different animal species from Bulgaria was as follows: dogs—21.1%; cats—17.7%; horses—8.3%; cattle—7.7%; sheep—32.2%; and goats—24.4%. We found the lowest overall HEV seropositivity in Plovdiv district (6.2%; 4/64; p = 0.203) and Smolyan district (8.8%; 4/45; p = 0.129), vs. the highest in Pazardzhik district (21.6%; 29/134; p = 0.024) and Burgas district (28.8%; 26/90; p = 0.062). To the best of our knowledge, this is the first serological evidence of HEV infection in dogs, cats, horses, cattle, sheep, and goats from Bulgaria. We found high HEV seropositivity in small ruminants (sheep and goats), moderate seropositivity in pets (dogs and cats), and a low level of seropositivity in large animals (horses and cattle). Previous Bulgarian studies and the results of this research show that HEV infection is widespread among animals in our country. In this regard, the Bulgarian health authorities must carry out increased surveillance and control of HEV infection among animals in Bulgaria. [ABSTRACT FROM AUTHOR]

Background: The aim of this research was to perform a seroepidemiological survey of hepatitis E virus (HEV) by an alternative method—serological establishment of the virus from pig liver transudate. This is the first such research in Bulgaria. Moreover, no studies have been conducted on HEV in pig liver in our country. Materials and Methods: A total of 90 liver samples from healthy pigs were collected from slaughterhouses in three parts of the country (Sofia, Lovech, and Stara Zagora districts). An equal number of samples (n = 30) were obtained from each district. Results: The overall HEV seropositivity was 67.7% of all 90 tested pig liver transudate samples; in Sofia district 20.0%, in Lovech district 90.0%, and in Stara Zagora district 93.3%. Conclusion: These data are similar to our previous serological studies for HEV in serum samples from industrial pigs, wild boars, and East Balkan swine. In this regard, testing the liver transudate could be a fine alternative method for seroepidemiological survey on HEV in swine. [ABSTRACT FROM AUTHOR]

The hepatitis C virus (HCV)-related bovine hepacivirus (BovHepV) can cause acute as well as persistent infections in cattle. The true clinical relevance of the virus is not yet known. As reliable antibody detection methods are lacking and prevalence studies have only been conducted in cattle and few countries to date, the true distribution, genetic diversity, and host range is probably greatly underestimated. In this study, we applied several RT-PCR methods and a nano-luciferase-based immunoprecipitation system (LIPS) assay to analyze bovine serum samples from Bulgaria as well as wild ruminant sera from Germany and the Czech Republic. Using these methods, BovHepV infections were confirmed in Bulgarian cattle, with viral genomes detected in 6.9% and serological reactions against the BovHepV NS3 helicase domain in 10% of bovine serum samples. Genetic analysis demonstrated co-circulation of highly diverse BovHepV strains in Bulgarian cattle, and three novel BovHepV subtypes within the genotype 1 could be defined. Furthermore, application of a nested RT-PCR led to the first description of a BovHepV variant (genotype 2) in a wild ruminant species. The results of this study significantly enhance our knowledge of BovHepV distribution, genetic diversity, and host range. [ABSTRACT FROM AUTHOR]

Information on hepatitis E virus (HEV) strains circulating in animal reservoirs in Bulgaria is currently lacking. Herein, by screening HEV seropositive sera obtained from Bulgarian swine and wild boars, viral RNA was detected at high prevalence rate (28.2%) in industrial pigs. Sequence analysis of the partial polymerase (RdRp) region revealed the highest genetic correlation with HEVs of genotype (Gt) 3 identified in French and Dutch patients. For three such strains, a 700-bp fragment of the open reading frame 2 gene was generated. On phylogenetic analysis, the Bulgarian strains clustered tightly (93.8–98.3% nt) with human and animal HEVs classified within the Gt3 subtype c. [ABSTRACT FROM AUTHOR]

Scientific evidence indicates that hepatitis E virus (HEV) infection is a zoonotic disease. Domestic pigs and wild boars are the main animal reservoir for HEV worldwide. The aim of this study was to analyze the seroprevalence of HEV infection among wild boars in western Bulgaria. Serum samples from 240 wild boars from two regions of the country (northwestern and southwestern) were tested for anti-HEV Immunoglobulin G (IgG) antibodies. The overall HEV seroprevalence was 40.8% (98/240); northwestern region 40.0% (48/120); southwestern region 41.7% (50/120). HEV seropositivity in the southwestern region was higher than in the northwestern region: odds ratio = 1.071 (95% confidence interval: 0.640-1.793). This research provided the first seroprevalence study to HEV in wild boars from western Bulgaria. [ABSTRACT FROM AUTHOR]

Lyme borreliosis, granulocytic anaplasmosis and monocytic ehrlichiosis are well studied in humans and dogs. In horses, these diseases are not widely investigated and limited information is available about their occurrence. The purpose of this study was to present the first ELISA-based report on the seroprevalence of Anaplasma phagocytophilum, Ehrlichia spp. and Borrelia burgdorferi in horses from Northern Bulgaria. A total of 192 horses were investigated from three regions in Northern Bulgaria (Northwestern, North-Central and Northeastern Bulgaria). All equine sera were tested for A. phagocytophilum, Ehrlichia spp. and B. burgdorferi antibodies by a commercial rapid ELISA test. Antibodies against A. phagocytophilum were found in all the three regions at a mean frequency of 12% (23/192), ranging from 9.38 to 15.63% by region. Antibodies against Ehrlichia spp. were found in horses from one region (Northeastern) at a rate of 0.5% (1/192). Anti-B. burgdorferi antibodies were detected in all the three regions with a mean frequency of 15.1% (29/192), ranging from 14.06 to 17.19% by region. A co-exposure to A. phagocytophilum and B. burgdorferi was observed in 6.3% of the cases (12/192). This is the first report on the natural exposure of horses to these bacteria (A. phagocytophilum, Ehrlichia spp. and B. burgdorferi) in Northern Bulgaria. [ABSTRACT FROM AUTHOR]

Hepatitis E virus (HEV) infection is widespread among domestic pigs, industrial swine, and wild boars in Bulgaria. The aim of the current research was to present the HEV seroprevalence among blood donors in Bulgaria. In the present study, 555 blood donors (479 males and 76 females) were enrolled from five districts in the country (Shumen, Pleven, Stara Zagora, Plovdiv, and Sofia districts). All blood samples were tested for anti-HEV IgG using the recomWell HEV IgG ELISA test (Mikrogen GmbH, Neuried, Germany). Each participating donor completed a short, structured, and specific questionnaire to document data on the current study. Anti-HEV IgG positive results were detected in 144 (25.9%) blood donors, including 129 (26.9%) males and 15 (19.7%) females. The established HEV seropositivity was 28.8% (23/80) in Shumen district, 23.2% (22/95) in Pleven district, 27.1% (38/140) in Stara Zagora district, 27.5% (44/160) in Plovdiv district, and 21.3% (17/80) in Sofia district. A high HEV seroprevalence was found for persons who declared that they were general hunters (48.7%; 19/39; p = 0.001) and hunters of wild boars (51.6%; 16/31; p = 0.001). We present the first seroprevalence rates of HEV infection in blood donors from Bulgaria. The results of our research showed high HEV seropositivity among blood donors. [ABSTRACT FROM AUTHOR]

The East Balkan swine (Sus scrofa) is the only aboriginal pig breed in Bulgaria, and it is indigenous to the eastern part of the country. The aim of the present study was to investigate East Balkan swine (EBS) in Bulgaria for serological evidence of hepatitis E virus (HEV). Sera from 171 swine from two parts of the country (northeastern and southeastern) were tested for anti-HEV IgG antibodies. The overall HEV seroprevalence was 82.5% (141/171), and for weaners it was 77.2% (44/57), for fattening pigs 79.0% (45/57), and for adults 91.2% (52/57). HEV positivity was higher in fattening pigs and adults compared to weaners: OR = 1.108 (95% CI: 0.456–2.692) and OR = 3.073 (95% CI: 1.016–9.294), respectively. This study provides the first evidence of exposure to HEV in EBS from Bulgaria. [ABSTRACT FROM AUTHOR]