Diagnostic Problem in a Child With an Isolated Renal Hydatid Cyst: A Case Report.

Cystic hydatid disease is a parasitic disease caused by the larvae of the small tapeworm Echinococcus granulosus. It is still a serious public health problem in endemic regions such as the Mediterranean basin, especially in the Balkans. Usually, the complaints caused by the cysts are non-specific and there are rarely abnormalities in routine laboratory tests. The most common is the involvement of the liver. The frequency of isolated kidney involvement, especially in a child, is uncommon. We describe a rare pediatric case of an isolated renal hydatid cyst presenting with a urinary tract infection-like clinical presentation, leading to misdiagnosis and delayed treatment.

Co-infection with plasmodium falciparum and COVID-19 with lethal outcome. First clinical case from Bulgaria.

The symptoms of COVID-19 include febrility and mainly catarrhal symptoms. In severe cases, patients present with progression to lower respiratory tract and acute respiratory distress (ARDS) and multi-organ dysfunction. Malaria caused by P. falciparum is a severe, endemic parasitosis, mainly in Africa. In some cases, it can be complicated with ARDS. We present a case of a patient who returned from Nigeria with respiratory symptoms, in which both COVID-19 infection and tropical malaria were proven; with a fatal outcome.

Autochthonous and Imported Visceral Leishmaniasis in Bulgaria-Clinical Experience and Treatment of Patients.

Visceral leishmaniasis (VL) is a severe endemic disease with a fatal outcome if left untreated. The symptoms of patients are diverse and atypical. Against the background of anemia and pancytopenia, the condition of the patients gradually worsens with marked cachexia. Through sharing our experience, we aim to draw attention to this deadly disease. Clinical and laboratory data for 58 patients with VL treated over a forty-five-year period are presented. The diagnosis was established within a duration of 1 to 28 months of illness. Continuous fever (38-42 °C), splenomegaly, hepatomegaly, severe anemia (decreased hemoglobin to lowest values of 31 g/L), leucopenia (lowest values of leucocytes et 0.5 g/L), and thrombocytopenia (reduced thrombocyte count to 29 g/L) were observed. The diagnosis was made on the basis of microscopic evidence of amastigote forms in bone marrow smears and serological tests. The patients were treated with Glucantime for 17 to 21 days. Relapses were observed in seven patients (12.1%) and fatal outcome was observed in two patients (3.5%) during treatment, who developed acute respiratory and cardiovascular failure. In Bulgaria, Visceral leishmaniasis is primarily endemic in the southern regions and should be suspected not only in patients who have returned from tropical and subtropical countries, but also in those who have not traveled abroad. The challenges associated with VL stem from delayed diagnosis of patients, as this disease remains unrecognized by physicians.

COVID-19 and gastrointestinal injury: a brief systematic review and data from Bulgaria.

In December 2019, a new Coronavirus (SARS-CoV-2) emerged in China, causing the pandemic disease COVID-19. The clinical presentation is variable, but the predominant symptoms are those of the upper respiratory tract. AIM: The aim of the current study is to describe the incidence and type of the gastrointestinal injury (GI) in COVID-19, as well as their prognostic value. MATERIALS AND METHODS: We conducted a coincidental search on this topic in PubMed, Web of Science and EMBASE. We also followed a group of 31 Bulgarian COVID-19 patients throughout the course of their disease and analyzed their symptoms (catarrhal and other) and outcome. RESULTS: The publications concerning our survey followed a total of 1509 COVID-19 patients. In the Bulgarian cohort, only 14 from the 31 patients were laboratory-confirmed COVID-19 cases. Approximately 1/3 of the infected individuals presented with GI. In some patients this was the first, or only, symptom of the disease. It was also indicative of a more severe disease course. CONCLUSION: GI may be an important symptom and prognostic factor in COVID-19. Therefore, patients with acute gastrointestinal symptoms must be actively tested for SARS-CoV-2.

Several Cases of Ocular Dirofilariasis in Bulgaria.

OBJECTIVES: In this study, we describe ocular dirofilariasis in Bulgaria. MATERIALS AND METHODS: We studied 7 patients with a subconjunctival or periorbital form of Dirofilaria repens infection during 2010-2019. Morphological, serological, and paraclinical diagnostic methods were used. RESULTS: The patients (6 females, 1 male) were aged between 23 and 72 years. In 3 patients, subcutaneous nodules were detected in the area of the upper eyelid, in 1 patient the location was suborbital. In 3 other patients, a subconjunctival location was found. All patients were cured definitively by removal of the larva, without etiologic treatment. CONCLUSION: The most reliable and easily accessible diagnostic method is morphological analysis by microscopy of histological preparations of the parasite. In dirofilariasis, ocular location is the most common in humans, and it deserves special attention of clinicians.

Clinical and epidemiological descriptions from trichinellosis outbreaks in Bulgaria.

Bulgaria is one of European countries where trichinellosis continues to be regularly diagnosed and registered. The clinical and epidemiological features of 72 cases of trichinellosis associated with five outbreaks caused by Trichinella spiralis and Trichinella britovi between 2009 and 2011, are described. At hospital admission, patients were often initially treated with antibiotics, without any improvement. A range of signs and symptoms were recorded, including: myalgia, elevated temperature, arthralgia, difficulty with movement, facial oedema, conjunctival hyperaemia, ocular haemorrhages, diarrhoea, skin rash, headache, and fatigue. Due to the variable clinical course of the disease, the diagnostic process for trichinellosis is often complex and difficult. This means the diagnosis may be established late for an appropriate treatment, potentially leading to a severe course of the disease with complications. Laboratory abnormalities were expressed by marked eosinophilia (97.2%), leucocytosis (70.8%), elevated serum creatine phosphokinase levels (82%), and antibody-positive results by ELISA and indirect hemagglutination. Patients were treated with albendazole (Zentel) 10 mg/kg for 7-10 days. In two outbreaks, the aetiological agent was T. spiralis, in one outbreak T. britovi, and an unknown Trichinella species in the fourth outbreak. The sources of infection were domestic pigs, probably fed with scraps and offal of wild game. In one outbreak, T. spiralis was also detected in brown rats trapped close to where the pig had been raised in the backyard. These epidemiological factors are relevant in considering implementation of targeted control programmes.

Epidemiological factors associated with human cystic echinococcosis: a semi-structured questionnaire from a large population-based ultrasound cross-sectional study in eastern Europe and Turkey.

BACKGROUND: Cystic echinococcosis (CE) is a neglected parasitic zoonosis prioritized by the WHO for control. Several studies have investigated potential risk factors for CE through questionnaires, mostly carried out on small samples, providing contrasting results. We present the analysis of risk factor questionnaires administered to participants to a large CE prevalence study conducted in Bulgaria, Romania and Turkey. METHODS: A semi-structured questionnaire was administered to 24,687 people from rural Bulgaria, Romania and Turkey. CE cases were defined as individuals with abdominal CE cysts detected by ultrasound. Variables associated with CE at P < 0.20 in bivariate analysis were included into a multivariable logistic model, with a random effect to account for clustering at village level. Adjusted odds ratios (AOR) with 95% CI were used to describe the strength of associations. Data were weighted to reflect the relative distribution of the rural population in the study area by country, age group and sex. RESULTS: Valid records from 22,027 people were analyzed. According to the main occupation in the past 20 years, "housewife" (AOR: 3.11; 95% CI: 1.51-6.41) and "retired" (AOR: 2.88; 95% CI: 1.09-7.65) showed significantly higher odds of being infected compared to non-agricultural workers. "Having relatives with CE" (AOR: 4.18; 95% CI: 1.77-9.88) was also associated with higher odds of infection. Interestingly, dog-related and food/water-related factors were not associated with infection. CONCLUSIONS: Our results point toward infection being acquired in a "domestic" rural environment and support the view that CE should be considered more a "soil-transmitted" than a "food-borne" infection. This result helps delineating the dynamics of infection transmission and has practical implications in the design of specific studies to shed light on actual sources of infection and inform control campaigns.

Involvement of the gallbladder in the course of viral hepatitis A in childhood.

The course of acute hepatitis A (AHA) in childhood is most often benign. There are relatively few reports of gallbladder involvement during such infection and its consequences. We discuss the cases of 75 children (mean age = 4.5 years) with confirmed AHA. Clinical and laboratory evaluation upon admission were classical for AHA. Only two children had normal bilirubin levels and 38.7% had visible involvement of the gallbladder with wall thickening of >3 mm, while six met criteria for acute acalculous cholecystitis. There appears to be no clear relationship between thickening of the gallbladder wall and transaminase levels, but a significant relationship with total and direct bilirubin levels (P < 0.05). Ultrasonographic examination should therefore be carried out in each case of AHA and follow-up should be performed with greater caution, if there are changes in the gallbladder wall.

Epididymal Dirofilariasis in a Child: First Case Report from Bulgaria.

OBJECTIVE: To present case of a child with epididymal dirofil-ariasis. CLINICAL PRESENTATION AND INTERVENTION: An 11-year-old boy was admitted to the Clinic of Pediatric Urology for elective surgery treatment of epididymal cyst on the left side. After removal, the cyst was sent for histological examination. Microscopic examination of the histological slides revealed cross-sections of a nematode belonging to Dirofilaria spp., differentiated morphologically as D. repens. After surgery, the patient recovered completely. CONCLUSIONS: In most parts around the world, dirofilariasis is a rare and neglected infection. Nevertheless, the clinicians and pathologists must be informed about it.

Investigation of Salmonella enteritidis outbreak in four kindergartens.

In June 2018, in the city of Sofia, Bulgaria, 40 children from four different kindergartens suffered from salmonellosis caused by S. enteritidis. They were reported to have consumed food prepared and delivered by a private catering service. The patients had fever, diarrhea, and some had vomiting and abdominal pain. Sixteen of them were treated in hospital, and the other 24 received home treatment. Some of the outpatients received antibiotic treatment despite WHO recommendations. All 40 isolates were positive for O: D, H: gm and H: m, and were confirmed to be Salmonella enteritidis, respectively. Using conventional and molecular methods, such as serotyping, Multiplex-PCR and PFGE, it was confirmed that the strains were epidemiologically related. Based on molecular genetic methods, we established that the epidemic outbreak had a common origin: contaminated food delivered by a private catering service, which was consumed at all four kindergartens.

Prevalence of abdominal cystic echinococcosis in rural Bulgaria, Romania, and Turkey: a cross-sectional, ultrasound-based, population study from the HERACLES project.

BACKGROUND: Cystic echinococcosis is a neglected zoonotic infection that is distributed worldwide and prioritised by WHO for control efforts. The burden of human cystic echinococcosis is poorly understood in most endemic regions, including eastern Europe. We aimed to estimate the prevalence of abdominal cystic echinococcosis in rural areas of Bulgaria, Romania, and Turkey. METHODS: We did a cross-sectional ultrasound-based survey that recruited volunteers from 50 villages in rural areas of Bulgaria, Romania, and Turkey. These villages were in provinces with annual hospital incidence of cystic echinococcosis within the mid-range for the respective countries. All people who attended a session were allowed to participate if they agreed to be screened. Abdominal ultrasound screening sessions were hosted in public community structures such as community halls, primary health-care centres, schools, and mosques. Lesions were classified using an adapted WHO classification. We reported the prevalence of abdominal cystic echinococcosis adjusted by sex and age through direct standardisation, using the country's rural population as a reference. FINDINGS: From July 1, 2014, to Aug 3, 2015, 24 693 individuals presented to screening sessions and 24 687 underwent ultrasound screening. We excluded a further six indivduals due to missing data, leaving 24 681 people in our analysis. Abdominal cystic echinococcosis was detected in 31 of 8602 people screened in Bulgaria, 35 of 7461 screened in Romania, and 53 of 8618 screened in Turkey. The age and sex adjusted prevalence of abdominal cystic echinococcosis was 0·41% (95% CI 0·29-0·58) in Bulgaria, 0·41% (0·26-0·65) in Romania, and 0·59% (0·19-1·85) in Turkey. Active cysts were found in people of all ages, including children, and in all investigated provinces. INTERPRETATION: Our results provide population-based estimates of the prevalence of abdominal cystic echinococcosis. These findings should be useful to support the planning of cost-effective interventions, supporting the WHO roadmap for cystic echinococcosis control. FUNDING: European Union Seventh Framework Programme.

Multiplex PCR Assay for Identifi cation and Differentiation of Campylobacter jejuni and Campylobacter coli Isolates.

INTRODUCTION: Campylobacter spp. are important causative agents of gastrointestinal infections in humans. The most frequently isolated strains of this bacterial genus are Campylobacter jejuni and Campylobacter coli. To date, genetic methods for bacterial identification have not been used in Bulgaria. We optimized the multiplex PSR assay to identify Campylobacter spp. and differentiate C. jejuni from C. coli in clinical isolates. We also compared this method with the routinely used biochemical methods. AIM: To identify Campylobacter spp. and discriminate C. coli from C. jejuni in clinical isolates using multiplex PCR assay. MATERIALS AND METHODS: Between February 2014 and January 2015 we studied 93 stool samples taken from patients with diarrheal syndrome and identified 40 species of Campylobacter spp. in them. The clinical material was cultured in microaerophilic atmosphere, the isolated strains being biochemically diff erentiated (hydrolysis of sodium hippurate for C. jejuni, and hydrolysis of indoxyl acetate for C. coli). DNA was isolated from the strains using QiaAmp MiniKit (QIAGEN, Germany). Twenty strains were tested with multiplex PCR for the presence of these genes: cadF, characteristic for Campylobacter spp., hipO for C. jejuni and asp for C. coli. RESULTS AND DISCUSSION: The biochemical tests identified 16 strains of C. jejuni, 3 strains of C. coli, and 1 strain of C. upsaliensis. After the multiplex PCR assay the capillary gel electrophoresis confirmed 16 strains of C. jejuni, 2 strains of C. coli and 2 strains of Campylobacter spp. - because of the presence of the gene cadF. C. jejuni has the gene hipO, and it is possible that this gene may not be expressed in the biochemical differentiation yielding a negative reaction as a result. In comparison, we can conclude that the genetic differentiation is a more accurate method than the biochemical tests. CONCLUSION: The multiplex PCR assay is a fast, accurate method for identifi cation of Campylobacter spp. which makes it quite necessary in the clinical diagnostic practice.

Production and characterization of monoclonal immunoglobulin A antibodies directed against Salmonella H:g,m flagellar antigen.

Hybridomas were generated after intragastral immunization of BALB/c mice with live Salmonella suberu and subsequent fusion between isolated spleen lymphoblasts and myeloma cells. Three monoclonal antibodies (MAbs) of immunoglobulin A (IgA) isotype were selected and characterized. All of them were found to recognize the H:g epitope in enzyme-linked immunosorbent assay and immunoblotting but did not react with all H:g-expressing strains in slide agglutination test. All MAbs strongly agglutinated Salmonella enteritidis type strain and a large number of S. enteritidis clinical isolates. They were not bactericidal in the presence of complement. All hybridoma clones produced secretory IgA forms, which were found in the gastrointestinal tract of mice bearing hybridoma as a subcutaneous 'backpack' tumor or after intravenous application of purified MAbs. The IgA MAbs stability demonstrated in different tests together with their antigen specificity and strong agglutination ability make them a useful diagnostic tool for serotyping of Salmonella strains.