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Oral vaccination as a method of rabies eradication in the field was first started in Switzerland in 1978 and after 1984 several other EU countries followed this practice. Due to oral vaccination some European countries are now rabies-free in terrestrial animals. In Slovenia, after the first experimental oral vaccination and study of vaccination models from 1988 -1992, the spring-autumn campaigns have been carried out since 1995. The model of oral vaccination of wildlife requires 16-20 baits per km2 in the vaccination area. The baits were distributed by plane. They were dispersed from a height of 300-500 m. The aeroplanes' paths were 1000 metres apart. In the vaccination campaigns two vaccines were used. Lysvulpen®, produced by the Bioveta company at the Czech Republic, was laid down in the southwestern part of the country, and Fuchsoral®, produced by the German company Impfstoffwerk Dessau-Tornau, was placed in the eastern part of Slovenia. A rapid decline of rabies was evidenced from 1995 to 1999, when the oral vaccination program in the whole territory using the aircraft baits distributing system was practiced. In 1999, only 6 rabies cases were laboratory-confirmed, whereas in 1995, 1089 rabies cases were documented. Of the 14 rabies cases detected in 1998, 12 were found as an island in a circle with a radius of 30 km in the centre of the vaccinated area. In 2000 and 2001, rabies incidence increased again, so it was decided to change the baits distribution system in the year 2001. The vaccination by crossing flights in certain areas was introduced. In the next year (2002), after changing the vaccination strategy, positive cases rapidly dropped and only 15 cases in 2002, and 8 cases were found in 2003, near the non-vaccinated border with Croatia.

Rabies is a major zoonosis for which diagnostic techniques can only be performed in the laboratory. Laboratory techniques are preferably oriented on tissue removed from the cranium: hippocampus (Ammon's horn), cerebellum and the medulla oblongata or tissue liquids. Clinical observation may only lead to a suspicion of rabies. The only way to perform a reliable diagnosis of the disease is to identify the virus or some of its specific components using laboratory tests such as histological identification of characteristic cell lesions, immunochemical identification of rabies virus antigen and virus isolation. Serological tests are rarely used in epidemiological surveys but much more frequently in control of the vaccination programs (e.g. oral vaccination). Most commonly used serological tests are the virus neutralization test on cell culture (FAVN), virus neutralization in mice and ELISA.

Respiratory diseases often correspond to primary infections with different pathogens of cattle, causing heavy economic losses in young stock and breeding herds. Between 2012 and 2014, nasal swab samples were collected from twentyeight herds from 133 affected live cattle that were clinically suffering from symptoms of respiratory disease, pyrexia, cough, serous nasal and lacrimal discharge, increased respiratory rate, and breath sounds. Individual swab samples were tested in the laboratory using three commercial and one in-house real-time PCR methods, to detect nucleic acids of a total of ten different respiratory pathogens. Pasteurella multocida (P. multocida) was detected in 58.65% of samples, Mannheimia haemolytica (M. haemolytica) in 15.04%, while Mycoplasma bovis (M. bovis) and Histophilus somni (H. somni) were positive in 9.77% of nasal swab samples. Among viral pathogens, the highest prevalence (40.60%) was observed for bovine respiratory syncytial virus (BRSV), followed by bovine coronavirus (BCV) 12.03%, bovine para-influenza 3 (PI-3) 3.01%, and bovine viral diarrhea virus (BVDV) with 1.50% of positive samples. The less frequently detected viral pathogens were bovine herpes virus type 1 (BHV-1) and bovine adenovirus (BAdV) with 0.75% positive samples each. The new implemented molecular methods can be an important diagnostic tool for laboratories and farmers to improve the therapy, control, and prevention of respiratory disease in cattle herds. [ABSTRACT FROM AUTHOR]

The article presents a study on the infection of equine arteritis virus (EAV) on a stud farm. It provides details of the observation methodology to detect the infection in horses by virus neutralisation test (VNT) for EAV antibody detection and virus isolation and reverse transcription polymerase chain reaction (RT-PCR) test. It discusses the significance of the research in controlling EAV infection and production of non-carrier stallions in infected populations.

This article discusses a study which investigated the epidemiology of the virus causing viral haemorrhagic septicaemia (VHS). The study made a comparison of nucleotide sequences of nine VHSV isolates and sequences available in the GenBank database. Virus isolation, immunoperoxidase test, reverse transcriptase polymerase chain reaction (RT-PCR) and phylogenetic analysis confirmed the virus. The study concluded that direct nucleotide sequencing of RT-PCR products is a reliable tool for fast routine genotyping in laboratories.

Serum and oral swab samples were collected from a persistently infected cow and her calf in a two-month period to test usefulness of oral swab samples for the detection of bovine viral diarrhea virus. Nucleic acids of the virus were detected by two molecular methods: conventional gel-based RT-PCR and commercial real-time RT-PCR. The bovine viral diarrhea virus genome was detected in serum and oral swab samples on days 0,7,15,22,23,29,36,37,43,44,46,51,52,53, and 57. The dry cotton swabs showed a reduction of diagnostic sensitivity after three days when samples were stored at room temperature (+ 21 °C), but storage of oral swab samples at + 4 °C or in a freezer (< -1 5 °C) for at least 10 days had no negative impact on the detection of the virus. No reduction of diagnostic sensitivity was observed when oral swab samples were collected in tubes with a liquid virus transport medium. Oral swabs provide an easy, reliable and cost-effective sampling tool for identification of PI animals, together with RT-PCR methods. The oral swab sampling could be especially useful for screening newborn calves during testing and removing PI animals from bovine viral diarrhea virus-infected herds. [ABSTRACT FROM AUTHOR]

Malignant catarrhal fever (MCF) is a ubiquitous disease of cattle and other ruminants caused by ovine herpesvirus-2 (OvHV-2) in Europe and other continents and alcelaphine herpesvirus-1(AlHV-1) in Africa. In March 2010 MCF was recognized in small private zoological garden in three American bison (Bison bison) imported from Vienna. They were housed in an enclosure next to llamas, goats and domestic sheep, with direct contact between all mentioned species. Typical clinical signs of the acute head and eye form of MCF were observed, with especially inflammation of conjunctives, oral and nasal mucosa and marked depression. Mortality was 100%. Necropsy findings in one bison were consistent with MCF. Acute ulcerative abomasitis and omasitis, acute hemorrhagic enterotyflitis, acute purulent bronchitis and moderate emaciation were found. The histopathology revealed mixed lymphohistiocytic and neutrophilic vasculitis in the brain, meninges, liver, spleen, heart, lungs and mixed lymphohistiocytic and neutrophilic hepatitis and adrenalitis. OvHV-2 DNA was detected post-mortem in tissue samples by PCR. Direct sequencing of the PCR product confirmed 100% nucleotide identity to OvHV-2 strain BJI035 and closely related to two OvHV-2 positive sheep samples collected in 2007. These are the first case of MCF confirmed by laboratory diagnostic methods in Slovenia. Furthermore, these are also the first cases of MCF in American bison in Slovenia. [ABSTRACT FROM AUTHOR]

This research describes the detection of six honeybee viruses in samples of clinically affected Carniolan gray bee collected between 2007 and 2009 on the territory of Slovenia. Using one-step reverse transcription-PCR (RT-PCR), 60 bee sam- ples originated from 45 apiaries were screened forthe presence of six honeybee viruses. Samples were found positive for acute bee paralysis virus (ABPV = 40%), black queen cell virus (BQCV = 83,3%), chronic bee paralysis virus (CBPV = 18,3%), deformed wing virus (DWV = 70%), Kashmir bee virus (KBV = 1,7%) and sacbrood bee virus (SBV = 8,3%). Mortality and paralysis were often evident in the apiaries and could be connected with ABPV and/or CBPV infections. Both viruses were detected in clinically af- fected apiaries with high bee mortality and with paralysis symptoms showing flightless bees, trembling and crawling at the hive entrance. The severity of clinical manifestation with high bee losses were associated with higher number of viruses detected in the samples. Among virus positive samples, 27% of them were infected with one virus, 30% with two viruses, 25% with three viruses and 15% of samples contained four viruses simultaneously. The results of this study provide data about the detection of several bee viruses in affected bee colonies and viral infections in Carniolan bee have to be investigated byfurther research. [ABSTRACT FROM AUTHOR]

The detection of infectious hematopoietic necrosis virus (IHNV) in infected rainbow trout Oncorhynchus mykiss and in cell culture supernatants stored under different conditions was studied. IHNV-positive fish visceral organ homogenates and cell culture supernatants were incubated at 4 and 25 degrees C. Virus titre was measured by virus isolation on epithelioma papulosum cyprini (EPC) cells and the IHNV RNA was detected by RT-PCR and semi-nested RT-PCR. The influence of repeated freezing and thawing on the virus isolation from organ homogenates and from cell culture supernatants was studied as well. It was possible to isolate the virus from IHNV-positive organ material during the 3 d of incubation at 4 degrees C but, only on the first day of incubation at 25 degrees C. Viral RNA could be amplified during the incubation period of 35 d at 4 degrees C but only during 8 d of incubation at 25 degrees C. In IHNV-infected cell culture supernatant stored at 4 degrees C, it was possible to detect virus for 36 and 16 d in supernatant stored at 25 degrees C. Viral RNA could be followed by using molecular methods during the entire experimental period of 123 d. Each cycle of freezing and thawing of samples resulted in a reduction of IHNV titre in the suspension of visceral organs, while the virus titre in cell culture supernatant remained almost the same following 33 freezing-thawing cycles. The present results show that rapid laboratory processing and storage of potentially virus-containing tissue samples as well as the use of different detection methods are very important for efficient IHNV diagnosis.