The prevalence of Coxiella burnetii in ticks and animals in Slovenia.

BACKGROUND: The obligate intracellular bacterium Coxiella burnetii causes globally distributed zoonotic Q fever. Ruminant livestock are common reservoirs of C. burnetii. Coxiella burnetii are shed in large numbers in the waste of infected animals and are transmitted by inhalation of contaminated aerosols. This study was conducted to evaluate the prevalence of C. burnetii infection in domestic animals and ticks in areas of Slovenia associated with a history of Q fever outbreaks. RESULTS: A total of 701 ticks were collected and identified from vegetation, domestic animals and wild animals. C. burnetii DNA was detected in 17 out of 701 (2.4%) ticks. No C. burnetii DNA was found in male ticks. Ticks that tested positive in the PCR-based assay were most commonly sampled from wild deer (5.09%), followed by ticks collected from domestic animals (1.16%) and ticks collected by flagging vegetation (0.79%). Additionally, 150 animal blood samples were investigated for the presence of C. burnetii-specific antibodies and pathogen DNA. The presence of pathogen DNA was confirmed in 14 out of 150 (9.3%) blood samples, while specific antibodies were detected in sera from 60 out of 150 (40.4%) animals. CONCLUSIONS: Our results indicate that ticks, although not the primary source of the bacteria, are infected with C. burnetii and may represent a potential source of infection for humans and animals. Ticks collected from animals were most likely found to harbor C. burnetii DNA, and the infection was not lost during molting. The persistence and distribution of pathogens in cattle and sheep indicates that C. burnetii is constantly present in Slovenia.

Hypertrophic Osteopathy Associated with Mycotic Pneumonia in a Roe Deer ( Capreolus capreolus).

Aspergillus fumigatus is one of the most common Aspergillus species causing disease in individual animals but it rarely affects deer species. We report a case of A. fumigatus infection causing mycotic pneumonia and hypertrophic osteopathy in a roe deer ( Capreolus capreolus).

Prevalence of Anti-Hepatitis E Virus Antibodies and First Detection of Hepatitis E Virus in Wild Boar in Slovenia.

Hepatitis E is an emerging zoonotic disease caused by hepatitis E virus (HEV). In this study, we investigated HEV presence in a wild boar (Sus scrofa) population of Slovenia. A total of 288 wild boar serum samples were collected throughout the country, and HEV infection was investigated by serology, using enzyme-linked immunosorbent assay (ELISA) and by HEV RNA detection using a real-time PCR assay. Antibodies against HEV were detected in 30.2% (87/288) of animals tested, whereas HEV RNA was detected in only one sample. This is the first evidence of HEV presence in the wild boar population in Slovenia, and these results suggest that these animals are part of the HEV epidemiological cycle in the country.

Mycobacterium spp. in wild game in Slovenia.

Wildlife species are an important reservoir of mycobacterial infections that may jeopardise efforts to control and eradicate bovine tuberculosis (bTB), caused by Mycobacterium bovis. Slovenia is officially free of bTB, but no data on the presence of mycobacteria in wild animals has been reported. In this study, samples of liver and lymph nodes were examined from 306 apparently healthy free-range wild animals of 13 species in Slovenia belonging to the families Cervidae, Suidae, Canidae, Mustelidae and Bovidae. Mycobacteria were isolated from 36/306 (11.8%) animals (red deer, roe deer, fallow deer, wild boar and jackal) and identified by PCR, commercial diagnostic kits and sequencing. Non-tuberculous mycobacteria identified in five species were Mycobacterium peregrinum, M. avium subsp. hominissuis, M. intracellulare, M. confluentis, M. fortuitum, M. terrae, M. avium subsp. avium, M. celatum, M. engbaekii, M. neoaurum, M. nonchromogenicum and M. vaccae.

Intestinal parasites of the red fox (Vulpes vulpes) in Slovenia.

In the present study, 428 foxes were collected and examined for intestinal helminths using the washing-out method. Parasites were found in 93.2% of the examined animals. The most frequently identified nematodes were Uncinaria stenocephala (58.9%), Toxocara canis (38.3%) and Molineus patens (30.6%). Other nematodes found were Pterygodermatites affinis (4.2%), Capillaria sp. (2.8%), Crenosoma vulpis (2.8%), Toxascaris leonina (2.5%), Trichuris vulpis (0.7%) and Physaloptera sp. (0.2%). Mesocestoides sp. (27.6%) and Taenia crassiceps (22.2%) were the most prevalent cestodes, followed by T. polyacantha (6.5%), Hymenolepis nana (2.1%), T. pisiformis (2.1%) and Dipylidium caninum (1.4%). The study also revealed four trematode species: Rossicotrema donicum (1.6%), Heterophyes heterophyes (1.1%), Metagonimus yokogawai (1.1%), Prohemistomum appendiculatum (0.4%) and two protozoan species: oocysts of Sarcocystis (2.8%) and Isospora (0.4%). This is the first extensive study on the intestinal parasites of the red fox (Vulpes vulpes) in Slovenia. The 2.6% prevalence of Echinococcus multilocularis in the same sample population as investigated herein has been reported previously (Vergles Rataj et al., 2010).

Evidence of Anaplasma phagocytophilum in game animals from Slovenia.

Anaplasma phagocytophilum is a tick-borne rickettsial pathogen responsible for granulocytic anaplasmosis in mammalian hosts including humans. Wild animals may play an important role in the epidemiology of this disease. The aim of this study was to estimate the prevalence of infection with A. phagocytophilum among wildlife in Slovenia. Serum samples (n = 376) from the most important game species [red deer (Cervus elaphus), roe deer (Capreolus capreolus), wild boar (Sus scrofa), chamois (Rupicapra rupicapra) and brown bear (Ursus arctos)] were examined by A. phagocytophilum-specific indirect fluorescent-antibody assay (IFA) and wild boar spleen samples (n = 160) were tested by polymerase chain reaction (PCR). A. phagocytophilum-specific antibodies were found in 72% of sera and A. phagocytophilum DNA was present in 6.2% of spleens. The data indicate that A. phagocytophilum is present and widespread in Slovenian game animals and that game species are involved in the natural life cycle of A. phagocytophilum.

A case of hepatic carcinoid in a red deer hind (Cervus elaphus).

A carcinoid tumour in the liver of a red deer hind (Cervus elaphus) is described. Macroscopically, the liver was considerably enlarged with multifocal, firm, yellow and red nodular neoplastic masses, which were histopathologically diagnosed as hepatic carcinoids. The diagnosis was confirmed by modified Grimelius staining, which demonstrated numerous small argyrophilic granules in the cytoplasm of neoplastic cells, and by immunohistochemistry. The neoplastic cells gave a strong positive reaction for neuron-specific enolase (NSE) and synaptophysin and a weak positive reaction for chromogranin A. To our knowledge, this is the first report of a hepatic carcinoid in red deer.

Detection of trace amounts of abamectin used as an antiparasitic agent in fallow deer tissues.

A sensitive and reliable method for the determination of trace amounts of abamectin in muscles, kidneys and fat tissue of fallow deer is presented. Abamectin was extracted from the tissues with acetonitrile and the extract was cleaned up on a C8 solid-phase extraction cartridge. Abamectin residue was derivatised with trifluoroacetic acid anhydride and 1-methylimidazole, and determined using reversed-phase high-performance liquid chromatography under isocratic conditions and fluorescence detection. The recoveries of the method were high and consistent, ranging from 78% to 90%. The limit of detection of the method was below 1 microg/kg when analysing muscle, kidney and fat tissue. Matrix-matched calibration was used in order to obtain accurate values and to avoid matrix interference.

Atmospheric flash injuries in roe deer (Capreolus capreolus).

The pathological and histopathological examination of two female roe deer found dead after a severe thunderstorm is described. One of the two roe deer was killed by a direct hit of atmospheric flash, while the other animal, lying 1.5 m from the first one, died as a consequence of ground current strike. The major part of the coat and skin of the animal that was killed by a direct hit was covered with scorched streaks. The trachea and bronchi of both animals contained a considerable amount of aspirated light-red foam. Necropsy revealed diffuse haemorrhages over the epicardium of the right atrium with a diameter of 1 millimetre. The liver and the kidney exhibited severe congestion.