The presence of malignant ovine babesiosis in Bosnia and Herzegovina indicates a possible emerging risk for Balkan region.

Malignant ovine babesiosis or ovine babesiosis caused by Babesia ovis is an important hemoprotozoan tick-borne disease of sheep. After 70 years, a study was conducted in Bosnia and Herzegovina, in the geographical areas of Podrinje and Eastern Herzegovina, aiming to molecularly identify and determine the presence of B. ovis infection in the sheep population. From 2019-2021, a clinical and molecular study was conducted in 53 flocks, and 192 sheep were examined, divided into two groups: clinically suspected and asymptomatic sheep. The presence of B. ovis was confirmed by PCR in blood samples of 75 % and 11.21 % of clinically affected and asymptomatic sheep, respectively. The majority of clinical cases of malignant ovine babesiosis were confirmed in the Rudo epidemiological unit (78.78 %) within the Podrinje region indicating typical seasonal pattern of disease occurrence and endemic focus. Rhipicephalus bursa was only tick species identified in Podrinje and Eastern Herzegovina. Acute form of disease was observed with dominant clinical signs of depression, fever, loss of appetite and respiratory distress. Fatal outcome was recorded in 12.28 % of cases. This study describes the molecular detection of B. ovis in sheep in Bosnia and Herzegovina for the first time. To the authors knowledge, this survey represents a report of the highest number of clinical cases of malignant ovine babesiosis in Europe. Since the disease has been recorded in the south-eastern border of Bosnia and Herzegovina, there is a high risk of disease transmission to a wider area of the Balkan region.

First confirmation of salmonid alphavirus infection in rainbow trout in Bosnia and Herzegovina and Serbia.

Sleeping disease is a highly infectious viral disease caused by salmonid alphavirus subtype 2 (SAV2 FW), affecting mainly rainbow trout Oncorhynchus mykiss farmed in freshwater. During March to May 2014, disease episodes with clinical signs of sleeping disease in rainbow trout fingerlings occurred almost simultaneously in 2 trout farms located in Bosnia and Herzegovina (BiH) and Serbia. The infection of rainbow trout with SAV2 FW in 2 farms was confirmed by virus isolation and molecular methods. This is the first isolation and molecular characterization of SAV2 FW in BiH and Serbia.

Serologic and molecular evidence for circulation of Crimean-Congo hemorrhagic fever virus in ticks and cattle in Bosnia and Herzegovina.

Crimean-Congo haemorrhagic fever virus is a tick-borne zoonotic agent of the genus Orthonairovirus, and the causal agent of Crimean-Congo haemorrhagic fever in humans. The study is aimed at investigating the presence of CCHFV RNA in ticks and cattle in Bosnia and Herzegovina (B&H), as well as the exposure of cattle to CCHFV in this country. Only one pool of three male Hyalomma marginatum ticks tested positive for viral RNA (3/760, 0.39%). No viral RNA was detected in 206 bovine blood samples using real-time reverse-transcriptase PCR (RT-qPCR). The overall seroprevalence of CCHFV in the cattle population was estimated as 14.97% (72/481). The results indicate the presence and the established circulation of CCHFV in B&H. Considering the endemic character of CCHFV in the Balkans as well as our results, the possibility that CCHFV is endemic in the territory of Bosnia and Herzegovina is considered to be very high.

First evidence of Crimean-Congo haemorrhagic fever virus circulation in Bosnia and Herzegovina.

BACKGROUND: Crimean-Congo haemorrhagic fever (CCHF) is a widespread tick-borne zoonosis with reported detection of virus and/or virus-specific antibodies from over 57 countries across Africa, Asia, Europe and the Middle East and is endemic in the Balkans. Detection of Crimean-Congo Haemorrhagic Fever Virus (CCHFV) antibodies in domestic ruminants has been important in providing initial evidence of virus circulation and in localising CCHFV high-risk spots for human infection. OBJECTIVES: The present study investigated the possible exposure of sheep to CCHFV in Bosnia and Herzegovina (B&H). METHODS: To investigate the presence of anti-CCHFV antibodies in sheep, all sera (n = 176) were tested using multi-species double antigen enzyme-linked immunosorbent assay (ELISA). Reactive sera were further complementary tested by adapted commercial indirect immunofluorescence assay (IFA) using FITC-conjugated protein G instead of anti-human immunoglobulins. RESULTS: CCHFV specific antibodies were detected in 17 (9.66%) animals using ELISA test. All negative sera were determined as negative by both tests, while 13 out of 17 ELISA-positive reactors were also determined as unambiguously positive by IFA test. The age group with the highest proportion of seropositive rectors were the oldest animals. CONCLUSIONS: This is the first report of anti-CCHFV antibodies in sheep from B&H providing the evidence of CCHFV circulation in the country's sheep population. So far, these findings indicate the circulation of the virus in the westernmost region of the Balkans and point to the potential CCHFV spread further out of this endemic area.

An update of the Culicoides (Diptera: Ceratopogonidae) checklist for the Balkans.

BACKGROUND: The prime significance of species belonging to the genus Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) is their ability to transmit viruses such as bluetongue virus (BTV) to wild and domestic ruminants. Prior to 1998, BTV was considered exotic in Europe, but according to recent history of its outbreaks, it has become endemic in southern and eastern European countries circulating beyond its expected historical limits, into the Balkan region. The wind-borne long-distance dispersal of Culicoides spp. over water bodies and local spreading between farms emphasize the necessity of filling in the information gaps regarding vector species distribution. In most Balkan countries, data on Culicoides fauna and species distribution are lacking, or information is old and scarce. RESULTS: During this study, 8586 specimens belonging to 41 species were collected. We present the first faunistic data on Culicoides species in the former Yugoslav Republic of Macedonia (FYROM), Kosovo, Montenegro and Serbia. For other countries (Bosnia and Herzegovina, Bulgaria and Croatia), all historical records were compiled for the first time and then expanded with our findings to various extents. In all countries, confirmed or suspected BTV vector species belonging to the subgenera Avaritia and Culicoides were collected. The total number of species sampled during our field collections was 20 in Bosnia and Herzegovina (15 new records), 10 in Bulgaria (2 new records), 10 in Croatia (5 new records), 13 in FYROM, 9 in Kosovo, 15 in Montenegro, and 28 in Serbia. Of these, 14 species were registered for the first time in this part of the Balkans. CONCLUSIONS: This paper provides the first data about Culicoides fauna in FYROM, Kosovo, Montenegro and Serbia, as well as new records and an update on the checklists for Bosnia and Herzegovina, Bulgaria and Croatia. These findings provide preliminary insights into the routes of BTV introduction and spreading within the Balkans, and present a valuable contribution to further research related to Culicoides-borne diseases in Europe.

First molecular detection of Babesia canis in dogs from Bosnia and Herzegovina.

Babesia spp. are tick-transmitted protozoan haemoparasites of great economic, veterinary and medical impact worldwide. Herein we reported the very high prevalence of autochthonous babesiosis in symptomatic dogs from Bosnia and Herzegovina in the period from 2014 to 2016. Eighty dogs that did not leave the country were examined using parasitological and molecular analyses and babesiosis was diagnosed in 82.5% and 85.0% of them, respectively (p < 0.001). One species, Babesia canis was identified using molecular methodology (PCR and sequence analysis). Statistical analyses showed that epizootiological characteristics have no influence on the possibility of infection. Agglomerative hierarchical clustering (AHC) analyses used for comparing the symptoms and clinical signs of infection in dogs pointed out that a high degree of anemia, followed by thrombocytopenia (89%), lethargy (100%), loss of appetite (95%), fever (66%) and icterus (61%) was dominant. In addition, results of the statistical analysis performed showed that more dogs with no data of tick prophylaxis (70%) were found Babesia infected. Those results point to further intensified epizootic surveys in the territory of Bosnia and Herzegovina.

Culicoides obsoletus (Diptera: Ceratopogonidae) in Bosnia and Herzegovina-first report.

The first occurrence of bluetongue disease in Bosnia and Herzegovina was registered in 2002 in the area of Kalesija municipality. Entomological investigation of the presence of Culicoides species in that area was conducted in 2007. The aim of the research was to establish the presence of the main vector of bluetongue virus. Collections and analyses of Culicoides midges were performed in accordance with the protocols of the National Reference Centre for Exotic Diseases (Centro Studi Malattie Esotiche) in Teramo, Italy. Traps for capturing midges were placed next to four sheep farms. During the investigation, a total of 2,256 Culicoides midges were collected and only one species was identified, Culicoides obsoletus Meigen, 1818.