First evidence of Brucella ovis infection in rams in the Pirot Municipality, Serbia.

This paper describes a research on Brucella ovis infection in rams in the Pirot Municipality of South Serbia. A positive result with indirect immunoenzyme test (i-ELISA) was confirmed in 67 (29.8%) and suspicious in 31 (13.8%) out of 225 tested rams. Complement fixation test (CFT) was used as a confirmation test on 67 ELISA positive sera and gave positive reaction in 41 (61.2%) ram serum samples. Rams originated from 113 flocks with 4751 sheep, from 28 villages in the Pirot Municipality of southern Serbia. Clinical examination was performed on epididymis and testes of 12 rams from 7 seropositive flocks by inspection and palpation. The examination showed scrotum asymmetry and unilateral increase of the epididymistail in 5 (41.7%) out of 12 seropositive rams. Pathomorphological examination of testes and epididymis confirmed pathological changes in 7 (58.3%) of the 12 examined rams. Onesided epididymitis with pronounced hypertrophy of the epididymitis was also confirmed. Twelve rams were tested for the presence of bacteria, i.e. 21 epididymis, testes and lymph nodes samples. We isolated 20 Brucella strains from 11 (91.7%) of the 12 examined animals. All isolates were identified with bacteriological and molecular techniques as B. ovis. This is the first evidence of ovine epididymitis (B. ovis) in Republic of Serbia.

A case of human monocytic ehrlichiosis in Serbia.

INTRODUCTION: Ehrlichiosis is a bacterial zoonosis transmitted by hematophagous arthropods--ticks. In humans, it occurs as monocytic, granulocytic, and ewingii ehrlichiosis. Pathological process is based on parasitic presence of Ehrlichia organisms within peripheral blood cells--monocytes and granulocytes. CASE OUTLINE: Fifty-two year old patient was admitted to hospital due to high fever of over 40 degrees C that lasted two days, accompanied with chills, muscle aches, malaise, loss of appetite, headache, confusion, breathing difficulties, and mild dry cough. The history suggested tick bite that occurred seven days before the onset of disease. Doxycycline was introduced and administered for 14 days, causing the disease to subside. Indirect immunofluorescence assay was used to analyze three serum samples obtained from this patient for Ehrlichia chaffeensis antibodies, and peripheral blood smear was evaluated for the presence of Ehrlichia and Ehrlichia aggregation into morulae. CONCLUSION: Ehrlichiosis should be considered in each case where there is a history of tick bite together with the clinical picture (high fever, chills, muscle aches, headache, generalized weakness and malaise, and possible maculopapular rash). The presence of Ehrlichia chaffeensis antibodies was confirmed in a patient with the history of tick bite, appropriate clinical picture and indirect immunofluorescence assay. This confirmed the presence of human monocytotropic ehrlichiosis, a disease that is uncommonly identified in our country.

Simultaneous detection of vaccinal and field infectious bursal disease viruses in layer chickens challenged with a very virulent strain after vaccination.

Infectious bursal disease virus is an important poultry pathogen. It is distributed worldwide and causes significant economic losses. In this study, a system was adopted for the simultaneous monitoring of vaccine and virulent strains using reverse transcription polymerase chain reaction (RT-PCR). After the decay of maternal antibodies, chickens were vaccinated at the age of 37 days with a virus of intermediate virulence and challenged at 5, 10 and 14 days post vaccination (dpv). The challenge was done with IBDV strain CH/99. Sequencing of the hypervariable region of VP2 has shown that CH/99 belongs to the very virulent group of viruses. The vaccine virus could be found in the bursa of Fabricius, spleen, thymus and bone marrow until 24 dpv. The CH/99 challenge virus was found in the bursa and lymphoid organs when chickens were challenged at 5 and 10 dpv. When challenge was performed at 14 dpv, the pathogenic virus could not be found in the bursa and other lymphoid organs.

Oropharyngeal tularemia in father and son after consumption of under-cooked rabbit meat.

Tularemia has been recognized for more than 10 y in Serbia, since the first epidemic of tularemia occurred in Sokobanja region in 1999. We report 2 cases of oropharyngeal tularemia in a father and son after the consumption of under-cooked rabbit meat. Both presented with fever, unilateral tonsillopharyngitis and cervical lymphadenitis.

Results of bacteriological and cytological examinations of the endometrium of subfertile mares in stud farms in Serbia.

Uterine microbiology, antimicrobial susceptibility and endometrial cytology were investigated in a total of 51 mares with fertility problems from 16 different stud farms in Serbia. Uterine cultures were performed after collection with a double guarded uterine swab, and endometrial cytology was evaluated after collection of endometrial cells with a special device (cytology brush). In 21 of 51 mares, at least one bacterial species was isolated from the uterus; the most frequent were Streptococcus equi subsp. zooepidemicus (13 isolates) and E. coli (four isolates). All isolates of Streptococcus equi subsp. zooepidemicus were susceptible to penicillin. Results from endometrial cytology were inconsistent; in 17 animals with positive bacteriological culture, cytology was not altered. It can be concluded that in Serbia, as in many other contries, Streptococcus equi subsp. zooepidemicus is the main cause for equine endometritis. It can be easily diagnosed by uterine culture but endometrial cytology does not always prove the existence of an endometrial infection with this agent.

Prevalence of antibodies to Rickettsiae in different regions of Serbia.

We assayed the presence of antibodies specific for Rickettsia typhi, R. akari, and R. conorii in sera of persons from several localities in Serbia with different geographic, climatic, and lifestyle characteristics. Sera from 140 patients with unclear clinical symptoms and 273 healthy persons were tested for the presence of rickettsiae-specific antibodies by indirect immunofluorescence assay. In this study, for the first time we detected the presence of rickettsiae from the spotted fever group in Serbia. We detected the presence of antibodies against R. conorii in the samples from all tested localities. The proportion of positive cases was low in the plain agricultural areas but reached up to 23% in the mountain areas. We also observed a significant number of cases positive for antibodies against R. akari. Antibodies specific for the antigens of R. typhi were detected in only 2 samples from the municipality of Pec (Kosovo region). These findings contribute to the prevalence of Rickettsia species in Southeast Europe. Our study also revealed a dramatic lack of awareness of rickettsioses among medical personnel and pointed to the need for urgent measures that would help improve the current situation in the region.

[Human ehrlichiosis].

BACKGROUND: Human ehrlichiosis is a newly recognized disease. It is a tick-borne disease caused by several bacterial species of the genhus Erlichia. These are small gram-negative pleomorphic cocci, that are obligatory intracellular bacteria. Tick Ixodes is the principle vector in Europe, and Amblyomma amenicanum in the United States. Bacterial organisms replicate in a tick, and are transmited from infected cells in a vector to the blood cells of animals or humans. Human ehrlichiosis is a name for a group of diseases caused by different species of Ehrlichia. One of them is the disease named human monocytic ehrlichiosis, caused by Ehrlichia chaffeensis, and the other is a human granulocytic ehrlichiosis caused by Anaplasma phagocytophilia. CASE REPORT: We reported a 23-year-old patient admitted for the clinical treatment with the symptoms of high febrility (above 40 degrees C), headache, vomiting, general weakness and exhaustion, but without data on a tick bite. The patient was treated with trimetoprim-sulfamethoxazole for a week when Ehrlichia chaffeensis was confirmed by the immunofluoroscence test, and the therapy contimed with doxacyclin. CONCLUSION: Human ehrlichiosis is also present in our country, so this disease should be considered everyday, especially in infectology practice.

Improved serodiagnosis of early Lyme borreliosis: immunoblot with local Borrelia afzelii strain.

To improve the serodiagnosis of Lyme borreliosis (LB) the performances of four tests were evaluated. An indirect immunofluorescent assay based on Borrelia burgdorferi s.s., enzyme-linked immunosorbent assay (ELISA) based on local isolates of Borrelia afzelii and B. burgdorferi s.s., and immunoblot (IB) of B. afzelii were prepared. The serum panels contained 214 serum samples: control group (n=120) and patients at different stages of LB (n=94). The specificity of IB was 96%, of in-house ELISA 93%, and of IFA 89%. In early LB the sensitivity of IFA was 36%, ELISA 67%, and IB 93%. In late-stage LB the sensitivity was: 72% for IFA, 80% for ELISA, and 94% for IB. Comparison of in-house and Behring ELISA showed that the sensitivity of the serological assay could be increased when the test was based on local Borrelia strains. IgM and IgG antibodies from sera of patients with early and late LB most frequently demonstrated reactivity to OspC. The other significant proteins in early LB were: p39, p41 in IgM IB, and p83/100, p39, Osp17 in IgG IB; in late LB: p39, p41 in IgM IB, and p83/100, Osp17, p21 and p43 in IgG IB. Using IB based on local B. afzelii isolates improves the serodiagnosis of early LB in our geographical region.

[An epidemic of tularemia in the Nish area]. / Epidemija tularemije u Niskom okrugu.

Tularaemia, as a rare disease, surprised medical workers in 1999 and 2000 when it was detected and diagnosed in 31 patients. In 1999 patients were registered in villages of the Sokobanja community, and in 2000 also in other communities in the district of Nish. Patients were peasants, mostly housewives. Glandular form of the disease was dominant. The main clinical symptoms were: tonsillitis, pain in the region of enlarged lymphatic glands and temperature, Biohumoral syndrome presented accelerated sedimentation of erythrocytes and elevated fibrinogen values. Antibody titer of Francisella tularensis was extremely high in the majority of patients. The disease was cured in all patients: in 27 patients after the first therapy, in 4 subjects after recidivation and 5 patients were surgically treated.