SUMMARY Based on morphological and genetic characteristics, we describe a new species of Hepatozoon in the European wild cat (Felis silvestris silvestris), herein named Hepatozoon silvestris sp. nov. The study also provides the first data on the occurrence of H. felis in this wild felid. Hepatozoon meronts were observed in multiple cross-sections of different organs of four (44%) cats. Additionally, extracellular forms, resembling mature gamonts of Hepatozoon, were found in the spleen and myocardium of two cats. Furthermore, tissues of six animals (67%) were positive by PCR. Hepatozoon felis was identified infecting one cat (11%), whereas the 18S rRNA sequences of the remaining five cats (56%) were identical, but distinct from the sequences of H. felis. Phylogenetic analyses revealed that those sequences form a highly supported clade distant from other Hepatozoon spp. Future studies should include domestic cats from the areas where the wild cats positive for H. silvestris sp. nov. were found, in order to investigate their potential role to serve as intermediate hosts of this newly described species. Identification of its definitive host(s) and experimental transmission studies are required for elucidating the full life cycle of this parasite and the possible alternative routes of its transmission.

In the past few years the interest of the scientific community on lungworms of the genus Troglostrongylus has grown due to the increased number of unexpected cases of infections with Troglostrongylus brevior in domestic cats from Mediterranean Europe, likely due to a spill-over from wild reservoirs. Thus, there is a merit to increase our knowledge on the occurrence of this parasite in felids from European regions. The present paper describes lung lesions associated with T. brevior infection in the endangered Eurasian lynx (Lynx lynx) from Bosnia and Herzegovina. The carcass of an illegally killed 3-year-old male Eurasian lynx was presented for necropsy at the Faculty of Veterinary Medicine of Sarajevo (Bosnia and Herzegovina). Grossly, multiple, multinodular, consolidated and firm, tan to grey areas, occupying the caudal third of caudal lung lobes, were observed. At cut section, the catarrhal fluid was draining from the airways. Larvae of T. brevior were found in tracheal scraping. The histopathological examination revealed multifocal to coalescing areas, centered on bronchi and bronchioles, and expanded alveoli filled with necrotic debris, degenerated inflammatory cells, mostly neutrophils and macrophages, and multiple cross sections of parasite larvae and thin-walled morulated eggs of lungworms. The paraffin-embedded lung samples were molecularly positive for T. brevior. This paper describes the first record of T. brevior in the Eurasian lynx and the associated host lung pathology. Given its pathogenic potential and the lack of data on troglostrongylosis in lynx populations, the occurrence and impact of Troglostrongylus spp. on wildlife health as well as the role of L. lynx as reservoir of infection for other felids, should be further investigated.

Red foxes (Vulpes vulpes) have recently been recognized as potential reservoirs of several vector-borne pathogens and a source of infection for domestic dogs and humans, mostly due to their close vicinity to urban areas and frequent exposure to different arthropod vectors. The aim of this study was to investigate the presence and distribution of Babesia spp., Hepatozoon canis, Anaplasma spp., Bartonella spp., ‘Candidatus Neoehrlichia mikurensis’, Ehrlichia canis, Rickettsia spp. and blood filaroid nematodes in free-ranging red foxes from Bosnia and Herzegovina. Spleen samples from a total of 119 red foxes, shot during the hunting season between October 2013 and April 2014 throughout Bosnia and Herzegovina, were examined for the presence of blood vector-borne pathogens by conventional PCRs and sequencing. In the present study, three species of apicomplexan parasites were molecularly identified in 73 red foxes from the entire sample area, with an overall prevalence of 60.8%. The DNA of B. canis, B. cf. microti and H. canis was found in 1 (0.8%), 38 (31.9%) and 46 (38.6%) spleen samples, respectively. In 11 samples (9.2%) co-infections with B. cf. microti and H. canis were detected and one fox harboured all three parasites (0.8%). There were no statistically significant differences between geographical region, sex or age of the host in the infection prevalence of B. cf. microti, although females (52.9%; 18/34) were significantly more infected with H. canis than males (32.9%; 28/85). The presence of vector-borne bacteria and filaroid nematodes was not detected in our study. This is the first report of B. canis, B. cf. microti and H. canis parasites in foxes from Bosnia and Herzegovina and the data presented here provide a first insight into the distribution of these pathogens among the red fox population. Moreover, the relatively high prevalence of B. cf. microti and H. canis reinforces the assumption that this wild canid species might be a possible reservoir and source of infection for domestic dogs.

BackgroundRed foxes (Vulpes vulpes) have recently been recognized as potential reservoirs of several vector-borne pathogens and a source of infection for domestic dogs and humans, mostly due to their close vicinity to urban areas and frequent exposure to different arthropod vectors. The aim of this study was to investigate the presence and distribution of Babesia spp., Hepatozoon canis, Anaplasma spp., Bartonella spp., ‘Candidatus Neoehrlichia mikurensis’, Ehrlichia canis, Rickettsia spp. and blood filaroid nematodes in free-ranging red foxes from Bosnia and Herzegovina.MethodsSpleen samples from a total of 119 red foxes, shot during the hunting season between October 2013 and April 2014 throughout Bosnia and Herzegovina, were examined for the presence of blood vector-borne pathogens by conventional PCRs and sequencing.ResultsIn the present study, three species of apicomplexan parasites were molecularly identified in 73 red foxes from the entire sample area, with an overall prevalence of 60.8%. The DNA of B. canis, B. cf. microti and H. canis was found in 1 (0.8%), 38 (31.9%) and 46 (38.6%) spleen samples, respectively. In 11 samples (9.2%) co-infections with B. cf. microti and H. canis were detected and one fox harboured all three parasites (0.8%). There were no statistically significant differences between geographical region, sex or age of the host in the infection prevalence of B. cf. microti, although females (52.9%; 18/34) were significantly more infected with H. canis than males (32.9%; 28/85). The presence of vector-borne bacteria and filaroid nematodes was not detected in our study.ConclusionThis is the first report of B. canis, B. cf. microti and H. canis parasites in foxes from Bosnia and Herzegovina and the data presented here provide a first insight into the distribution of these pathogens among the red fox population. Moreover, the relatively high prevalence of B. cf. microti and H. canis reinforces the assumption that this wild canid species might be a possible reservoir and source of infection for domestic dogs.

Abstract The aim of this study was to determine the occurrence and distribution of Cryptosporidium spp. and Giardia duodenalis among free-living red foxes (Vulpes vulpes) in Bosnia and Herzegovina. For this purpose, a total of 123 fecal samples from red foxes, shot during hunting seasons between January 2011 and March 2012 were examined using immunofluorescent microscopy. Overall, observed prevalences of Cryptosporidium spp. and Giardia duodenalis were 3.2 % (4/123) and 7.3% (9/123), respectively. The results show that foxes might play the role of potential reservoirs of Cryptosporidium and Giardia parasites, but further molecular analysis are necessary to elucidate the source of infection, routes of transmission and zoonotic potential of these two pathogens.

Thelazia callipaeda (Spirurida, Thelaziidae), also known as “oriental eyeworm”, is a small nematode parasite that lives in the conjunctival sac of domestic and wild carnivores, rabbits and even humans, causing mild (e.g., conjunctivitis, epiphora, and ocular discharge) to severe (e.g., keratitis, and corneal ulcers) ocular disease. This study reports, for the first time, the occurrence of T. callipaeda infection in the Balkan regions (i.e., Bosnia and Herzegovina and Croatia), it provides genetic evidence on the origin of the infection in that area and discusses potential expansion pathways in the near future. This survey was conducted in two Western Balkan countries, Bosnia and Herzegovina and Croatia. At necropsy, from January 2011 to April 2014, a total of 184 carcasses of red foxes were examined throughout the study area and worms were collected from the conjunctival sac. In the same period, worms were also collected during clinical examination from the conjunctival sac of four dogs and a cat from Bosnia and Herzegovina and two dogs from Croatia. All nematodes collected were morphologically identified and molecularly characterized by sequencing of partial cox 1 gene. T. callipaeda was observed in 51 (27.71%) foxes and the highest prevalence (50.0%) was in the region of East Bosnia. Beside the 4 cases of hyperemia (7.84%), most of the infected animals had no signs of ocular infection (n = 47, 92.15%). A total of 417 adult nematodes collected (364 from foxes, 51 from dogs, 2 from cat) were morphologically and molecularly identified as T. callipaeda haplotype 1. This is the first report of autochthonous cases of T. callipaeda infection in red foxes, dogs and cat in Bosnia and Herzegovina and Croatia and data presented here suggest that reports of thelaziosis in other Balkan areas are, as yet, not diagnosed most likely due to the lack of awareness of practitioners. In addition, data regarding the spread of the infection in Europe over the last ten years suggests that an increasing pattern in the distribution of this disease in domestic and wild animals should be expected in the future.

BackgroundThelazia callipaeda (Spirurida, Thelaziidae), also known as “oriental eyeworm”, is a small nematode parasite that lives in the conjunctival sac of domestic and wild carnivores, rabbits and even humans, causing mild (e.g., conjunctivitis, epiphora, and ocular discharge) to severe (e.g., keratitis, and corneal ulcers) ocular disease. This study reports, for the first time, the occurrence of T. callipaeda infection in the Balkan regions (i.e., Bosnia and Herzegovina and Croatia), it provides genetic evidence on the origin of the infection in that area and discusses potential expansion pathways in the near future.MethodsThis survey was conducted in two Western Balkan countries, Bosnia and Herzegovina and Croatia. At necropsy, from January 2011 to April 2014, a total of 184 carcasses of red foxes were examined throughout the study area and worms were collected from the conjunctival sac. In the same period, worms were also collected during clinical examination from the conjunctival sac of four dogs and a cat from Bosnia and Herzegovina and two dogs from Croatia. All nematodes collected were morphologically identified and molecularly characterized by sequencing of partial cox 1 gene.ResultsT. callipaeda was observed in 51 (27.71%) foxes and the highest prevalence (50.0%) was in the region of East Bosnia. Beside the 4 cases of hyperemia (7.84%), most of the infected animals had no signs of ocular infection (n = 47, 92.15%). A total of 417 adult nematodes collected (364 from foxes, 51 from dogs, 2 from cat) were morphologically and molecularly identified as T. callipaeda haplotype 1.ConclusionThis is the first report of autochthonous cases of T. callipaeda infection in red foxes, dogs and cat in Bosnia and Herzegovina and Croatia and data presented here suggest that reports of thelaziosis in other Balkan areas are, as yet, not diagnosed most likely due to the lack of awareness of practitioners. In addition, data regarding the spread of the infection in Europe over the last ten years suggests that an increasing pattern in the distribution of this disease in domestic and wild animals should be expected in the future.