Clinical Case of Life-threatening Co-infection Due to Dirofilaria Immitis and Aelurostrongylus Abstrusus in a Cat: First Report of Feline Heartworm Disease in Bulgaria.

The present report describes the first clinically manifested and serologically proven case of Dirofilaria immitis infection in a cat in Bulgaria. A 10-year-old intact male cat was referred to the Small Animal Clinic, Trakia University with a history of anorexia, weight loss, intermittent coughing and itching skin lesions on the head and neck. Physical examination revealed abnormal heart sounds and respiration, cyanosis of the mucous membranes, and generalized enlargement of the lymph nodes. Mild infestation with hard ticks and fleas was also detected during the initial skin inspection. In addition, adult Otodectes cynotis mites were observed in the skin lesions. The fecal sample was positive for larvae of Aelurostrongylus abstrusus and eggs of Toxocara cati. Blood serology revealed antigens of D. immitis as well as antibodies against both feline immunodeficiency virus and D. immitis. Thoracic radiographic findings included a pronounced generalized reticular interstitial pattern; alveolar and bronchial shades with multiple nodular thickenings throughout the lungs. Electrocardiography demonstrated a sinus tachycardia, a peaked P-wave (P-pulmonale) and an abnormally low ST-segment. The clinical signs disappeared after treatment with selamectin, doxycycline and corticosteroids. Despite the improvement in general health condition, the cat suddenly died several months later.

Preliminary Study on the Prevalence of Endoparasite Infections and Vector-borne Diseases in Outdoor Dogs in Bulgaria.

The present work was designed to evaluate the prevalence of gastrointestinal parasites and some vector-borne pathogens in dogs in Bulgaria. A total of 172 owned dogs, keeping outside, were included in the study. Fecal samples were examined using standard flotation and sedimentation methods. Blood samples were processed by Knott's technique, SNAP™ 4Dx Plus Test (IDEXX) and Angio Detect™ Test (IDEXX). The overall prevalence of gastrointestinal parasites was 64.5%. Eggs of hookworms (Ancylostoma sp. and Uncinaria sp.) were the most frequently detected (54.1%), followed by Trichuris vulpis (15.1%), Capillaria sp. (11.0%), Toxocara canis (6.4%), Cystoisospora sp. (4.1%), Sarcocystis sp. (2.3%), Toxascaris leonina (1.7%), Taenia sp. (1.2%) and Linguatula serrata (0.6%). In addition, hookworms were the most commonly involved in the cases of single infection (20.3%). Combinations between Capillaria sp./hookworms and T. vulpis/hookworms were the most common co-infections (4.1% and 2.9%, respectively). Blood samples revealed the presence of antibodies against Ehrlichia sp. (13.4%), Anaplasma sp. (13.4%) and Borrelia burgdorferi (1.7%). Antigens of Dirofilaria immitis and Angiostrongylus vasorum were detected in 10.5% and 0.6% of the samples tested, respectively. Microfilariae of Dirofilaria repens were found in 5.8% of the blood samples. Additionally, the prevalence of D. immitis and Ehrlichia sp. was significantly higher in adult than in young dogs (p<0.05). In contrast, the gender was not considered as a risk factor contributing to the occurrence of infections.

Multilocus sequence typing of canine Giardia duodenalis from South Eastern European countries.

Giardia duodenalis is a worldwide occurring protozoan that can infect various mammalian hosts. While living conditions are getting closer between pet animals and owners, there is discussion whether dogs may contribute to the transmission of these pathogens to humans. The present study was conducted in order to identify the Giardia assemblages in dogs from South Eastern Europe. For this purpose, 1645 faecal samples of household and shelter dogs from Albania, Bulgaria, Hungary, Macedonia, Romania and Serbia were tested for Giardia coproantigen by enzyme-linked immunosorbent assay (ELISA). A subset of 107 faecal samples demonstrating Giardia cysts by direct immunofluorescence assay (IFA) or microscopy (15-22 per country) plus 26 IFA-positive canine faecal samples from Croatia were used for DNA extraction and multilocus sequence typing with nested PCRs targeting five different gene loci: SSU rRNA, ITS1-5.8S-ITS2, beta giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi). One third (33.7%) of the samples tested positive for Giardia antigen in the coproantigen ELISA. Shelter dogs were infected more frequently than household dogs (57.2 vs. 29.7%, p < 0.01). Amplification was obtained in 82.0, 12.8, 11.3, 1.5, and 31.6%, of the investigated samples at the SSU rRNA, bg, gdh and tpi loci and the ITS1-5.8S-ITS2 region, respectively. The dog-specific assemblages C and D were identified in 50 and 68 samples, respectively. The results demonstrate that G. duodenalis should be considered as a common parasite in dogs from South Eastern Europe. However, there was no evidence for zoonotic Giardia assemblages in the investigated canine subpopulation.

Efficacy of parenteral administration of ivermectin in the control of strongylidosis in donkeys.

Investigations into the efficacy of parenteral ivermectin (Pandex) administration for strongylidosis control in donkeys were carried out. The preparation was applied subcutaneously at a dose of 0.2 mg/kg (1 ml/50 kg body weight). One day prior to the treatment and 14 days post-treatment, individual coprological samples were obtained for faecal nematode egg counts and larval culture. The study was performed on 263 donkeys originating from different regions of Bulgaria. Prior to the treatment and 20 days after that, blood samples were obtained from 64 previously infected animals for monitoring of changes in eosinophil leukocyte counts. The subcutaneous application of ivermectin had an efficacy of 96% in terms of reduction of faecal egg counts. In 92.2% of infected donkeys, a complete reduction of faecal eggs count occurred (0 eggs per gram of faeces epg), whereas in the remaining 7.8% of the infected donkeys, the egg counts were reduced by 72%. The reduction in faecal egg counts did not result in changes in eosinophil counts. The results obtained as well as the lack of local changes after the subcutaneous application of ivermectin in donkeys allow us to recommend its use for control of strongyles in donkeys.

An approach to modification of the repair activity in eucaryotic cells by biological factors.

This work was devoted to investigation or repair regulation by biological factors: viruses and interferon. DNA damage induced by gamma- and UV-irradiation, ethyleneimine and 4-nitro-quinoline-1-oxide (4-NQO) were studied, by sedimentation of lysed cells through alkaline sucrose gradients, by hydroxylapatite column chromatography and by the chromosomal aberration test. The reproducible vaccinia virus resulted in simulation repair activity of chick embryo cells after treatment with 4-NQO. Interferon, added after gamma- and UV-irradiation, decreased the chromosomal aberration level, stabilized it after ethyleneimine treatment and also stimulated the ability of cells to rejoin DNA breaks induced by 4-NQO. The cause of this phenomenon is discussed.

Molecular-genetic effects of cadmium chloride.

We studied the effect of cadmium chloride on: (1) the DNA of human cells; (2) the mutagenic effect of reproducing Kilham virus; (3) the synthesis of virus-induced interferon, and (4) the reproduction of oncogenic (mammalian leucosis) virus. Cadmium chloride caused degradation of DNA in human- and rat-embryo cells. Culture infected by the virus in the presence of cadmium sulphate had the highest yield of cells with chromosomal aberrations. Cadmium chloride caused marked inhibition of the virus-induced synthesis of interferon. The introduction of cadmium chloride into diploid cells infected by the leucosis virus caused a 3-4 fold increase in the yield of virus-induced transformation foci.