A case of mortality in a re-introduced European bison associated with severe pneumonia caused by Dictyocaulus viviparus.

BACKGROUND: Dictyocaulosis is a parasitic disease caused by pulmonary nematodes from genus Dictyocaulus affecting various ungulate hosts. It can cause verminous bronchopneumonia and for heavily infected individuals, fatal outcomes can occur. CASE PRESENTATION: The study describes the case of a male European bison which died three months after relocation from Slovakia to one of the reintroduction areas in Nucșoara village, Fagaraș Mountains, Romania. Necropsy revealed the presence of pulmonary nematodes in the respiratory tract. Morphology and molecular diagnosis was performed and Dictyocaulus viviparus was identified. CONCLUSION: Rigorous health monitoring is essential to assure the success of reintroduction programs, understanding and preventing infectious diseases and limiting their impact on population health.

Toxoplasma gondii in small mammals in Romania: the influence of host, season and sampling location.

BACKGROUND: Toxoplasma gondii is a protozoan parasite that infects a large spectrum of warm-blooded animals, including humans. Small rodents and insectivores play an important role in the epidemiology of T. gondii and may serve as a source of infection for both, domestic and wild definitive felid hosts. Factors influencing the occurrence of T. gondii in wild small mammals are unknown, despite the fact that many intermediate host species are identified. We have used small mammals (Rodentia and Lipotyphla) captured over two years in various habitats, both in urbanised and in natural landscapes. We assessed the importance of land-use, season and host ecology on T. gondii infection. RESULTS: We examined 471 individuals belonging to 20 small mammal species, collected at 63 locations spread over wide altitude, habitat and land-use ranges from Romania. Heart tissue samples were individually analysed by PCR targeting the 529 bp repetitive DNA fragment of T. gondii. The overall prevalence of infection was 7.3%, with nine species of rodents and two species of shrews being found to carry T. gondii DNA. Five species showed high frequency of infection, with the highest prevalence found in Myodes glareolus (35.5%), followed by Spermophilus citellus (33.3%), Sorex minutus (23.1%), S. araneus (21.7%) and Micromys minutus (11.1%). Adults seemed more often infected than young, however when controlling for season, the difference was not significant, as in spring both adults and young showed higher infection rates, but more adults were sampled. Contrary to our expectations, urban/rural areas (with their implicit high density of domestic feline presence) had no effect on infection prevalence. In addition, neither habitat, nor land-use at sampling sites was important as only geographical location and host species were contributing factors to the infection risk. CONCLUSIONS: High prevalence of T. gondii infection showed a highly localised, patchy occurrence, with long living and higher mobility host species being the most common carriers, especially during autumn.

Characterization of tongue worms, Linguatula spp. (Pentastomida) in Romania, with the first record of an unknown adult Linguatula from roe deer (Capreolus capreolus Linnaeus).

Specimens of the pentastomid parasite, Linguatula serrata, have been reported from several animals in Romania, including some domestic dogs translocated to other parts of Europe. In this study, gray wolves (Canis lupus, n = 80), golden jackals (C. aureus, n = 115), red foxes (Vulpes vulpes, n = 236), and roe deer (Capreolus capreolus, n = 1) were examined for pentastomes. Overall, 17.5% of wolves were found to be infected with specimens of Linguatula, with a range of infections of one to five individuals per animal. Golden jackals and foxes had much lower infection levels, with 1.73% of golden jackals and 1.69% of foxes infected; both host species were found to be infected with one or two individual pentastomes per animal. The single deer specimen was infected with three individual pentastomes. The pentastomes collected from the wolves and golden jackals were determined to be immature and mature adult specimens of L. serrata based on morphological examination and molecular analysis using the 18S rRNA gene. No pentastomes collected from the red foxes were available for identification. The pentastomes collected from the roe deer were expected to be L. arctica but determined to be mature adult male specimens of an unknown Linguatula, herein, referred to as Linguatula sp. based on its morphology; the results of molecular sequencing for the Linguatula specimen collected from the deer were inconclusive, preventing a final species identification. This study presents the first report of L. serrata in any hosts from Romania through both morphological and molecular characterization, and also presents the first report of a Linguatula sp. in Ca. capreolus, utilizing morphological characterization. Issues of morphological variability are discussed, including the presence of spines in the hook pit of specimens of Linguatula. This study highlights the need to examine all specimens of Linguatula to confirm the stage of development. Despite the inconclusive molecular result for some specimens, the authors still urge future researchers to incorporate a combined molecular and morphological approach in identifying specimens of Linguatula.

Ixodid ticks parasitizing wild carnivores in Romania.

In Romania, data regarding hard-tick diversity and tick-host associations in wild carnivores are scarce. We aimed to identify tick species in wild carnivores and to establish reliable data on tick-host associations. The study was conducted in various Romanian localities from all five ecoregions found in the country. Fourteen species of wild carnivores were examined. Immature and adult ticks were collected and identified using the morphological keys. The frequency and mean intensity of tick infestation, overall and differentiated by species, developmental stage and host were calculated. Of 202 wild carnivores, 68 were parasitized by seven tick species (predominantly Ixodes ricinus, I. hexagonus and Dermacentor reticulatus). The mean intensity of tick infestation was similar in males (6.97, BCa 95% CI 5.15-9.88) and females (5.76, BCa 95% CI 4.15-9.17). The highest prevalence of infested animals was recorded in the pannonian and steppic ecoregions, 66.7 and 52.7%, respectively. In the continental ecoregion the prevalence was 26.7%, whereas in the pontic ecoregion it was 28%. The lowest value, 16.7% was recorded in the alpine ecoregion. In total 430 ticks were collected, and 24.8% (n = 50) of the animals were infested with more than one tick species. Fourteen new tick-host associations were recorded. Our results suggest that anthropogenic changes of the environment lead to the diminishing of the boundaries, between wild and domestic animals, increasing the exposure for both animals and humans, to infective agents, including tick-borne pathogens.

Angiostrongylus chabaudi (Biocca, 1957) in wildcat (Felis silvestris silvestris, S) from Romania.

Angiostrongylus chabaudi is a rare cardio-pulmonary nematode infecting felids. Although almost 60 years have passed since the original description of the species in Italy, this parasite has been seldom found in domestic and wildcats in southern Europe. The present study aims to report a new case of patent A. chabaudi infection in a road-killed wildcat from Maramureș County in Northern Romania. The necropsy revealed the presence of parasites in the pulmonary arteries and the right ventricle, and the fecal examination showed the presence of L1 larvae. Parasites were morphologically and morphometrically characterized as A. chabaudi, showing 100 % nucleotide similarity to an Angiostrongylus sp. originating from a wildcat from Germany and 99 % to A. chabaudi from Italy. This study reports A. chabaudi for the first time in Eastern Europe, expanding knowledge about the distribution range of this species.

Laboratory development of Dermacentor marginatus ticks (Acari: Ixodidae) at two temperatures.

The influence of two temperatures on the development of Dermacentor marginatus evolutive cycle was studied. Tests performed under controlled laboratory conditions at 21 °C, 80 % RH and 27 °C, RH 80 %, on ten fully engorged female ticks collected from naturally infested goats, in Cluj County, Romania. Hatched larvae were fed on white mice and the nymphs and adults on guinea pigs. The following parameters were evaluated: egg incubation; pre-feeding, feeding and pre-moulting for larvae and nymphs; pre-feeding, feeding, pre-oviposition and oviposition for females. Significant differences (p < 0.001) were observed in: egg incubation period (29.4 ± 1.07 at 21 °C; 5.9 ± 0.73 at 27 °C) and pre-moulting duration of larvae (18.9 ± 1.02 at 21 °C; 6.1 ± 0.58 at 27 °C) and nymphs (21.3 ± 0.87 at 21 °C; 19.9 ± 0.71 at 27 °C). The average duration of the developmental cycle was 133.9 days (range 122-154 days) at 21 °C, and 94.2 days (range 83-111 days) at 27 °C.

Fleas of wild carnivores in Romania: diversity, distribution, and host-associations.

BACKGROUND: Fleas are important hematophagous insects, infesting mammals and birds with a worldwide distribution. Fleas of medical importance have been reported from various carnivores worldwide, such as felids, canids, or mustelids. Romania hosts a wide carnivore diversity, but very little is known about flea species that parasitize these animals in Romania. This study aimed to provide a better understanding of the fleas' diversity and their distribution in a relatively large and diverse number of wild carnivore hosts from Romania. METHODS: From 2013 to 2021, 282 carcasses of wild carnivores from different locations in Romania were collected and examined for the presence of ectoparasites. All collected fleas were morphologically identified using specific keys and descriptions. An analysis of the co-occurrence networks was performed. RESULTS: A total of 11 flea species were identified: Pulex irritans (41.09%), Paraceras melis (20.11%), Ctenocephalides felis (7.33%), Ctenocephalides canis (7.83%), Monopsyllus sciurorum (11.11%), Chaetopsylla trichosa (21.96%), Chaetopsylla homoea (5.5%), Chaetopsylla tuberculaticeps (100%), Chaetopsylla rothschildi (13.33%), Chaetopsylla sp. (14.34%), Chaetopsylla globiceps (5.12%), Echidnophaga gallinacea (10%). The statistical analyses showed a significant difference between the infestation of Martes foina with females being more frequently infected than males (66% versus 33%). Paraceras melis infesting Meles meles had a significantly higher prevalence in female badgers than in males (× 2 = 7.7977, P < 0.01) and higher intensities of infestations in males than in females (t = 1.871, P < 0.05). CONCLUSIONS: This is the first large-scale study investigating the distribution and diversity of flea species infesting wild carnivores in Romania. Three flea species were identified for the first time in Romania (E. gallinacea, C. homoea, and C. tuberculaticeps).

Diversity of Crenosoma species in mustelids with the first molecular characterization of C. melesi and C. petrowi.

Species of genus Crenosoma have a wide distribution and are reported in Europe, the Americas, and Asia. Currently, the genus includes 14 nominal species, out of which 9 are parasitic in mustelids. Two species are mostly reported in mustelids from Europe, namely C. melesi and C. petrowi. Up to now, no genetic sequences are deposited in GenBank for any of the two. The aims of this study were to investigate the distribution, prevalence, and diversity of Crenosoma spp. infecting mustelids in Romania and to genetically characterize the species. Mustelids (n = 247) were collected over a period of 7 years from different locations in Romania and the respiratory tract was removed and examined for nematodes. Detected nematodes were morphologically identified and fragments of two genes were sequenced. Sampled mustelids included Eurasian badger, Meles meles (n = 102), Eurasian otter, Lutra lutra (n = 20), beech marten, Martes foina (n = 36), European pine marten, Martes martes (n = 5), steppe polecat, Mustela eversmanii (n = 1), European mink, Mustela lutreola (n = 1), least weasel, Mustela nivalis (n = 2), European polecat, Mustela putorius (n = 78), and marbled polecat, Vormela peregusna (n = 1). Nematodes from Eurasian badgers were morphologically identified as C. melesi (n = 13, 12.74%) and C. petrowi (n = 3, 2.94%). Nematodes from the beech martens were identified as C. petrowi (n = 6, 16.66%), C. vulpis (n = 1, 2.78%) and Crenosoma spp. (n = 3, 8.33%). Co-infections with two Crenosoma species were detected in one beech marten (C. petrowi + C. vulpis, n = 1, 2.77%) and in one European pine marten [C. petrowi + C. vulpis (n = 1, 20%)]. Two genes of Crenosoma melesi and C. petrowi were partly sequenced for the first time. We report new host-parasite associations for M. martes and C. vulpis. However, further studies are needed in order to determine the host-parasite associations and to improve the understanding of the epidemiology of Crenosoma nematodes.

Diversity of Thelazia spp. in domestic cattle from Romania: epidemiology and molecular diagnosis by a novel multiplex PCR.

BACKGROUND: Thelaziosis is a neglected vector-borne disease caused by parasitic nematode worms of the genus Thelazia which affects various hosts. Limited attention has been given to ungulate-associated Thelazia species. Current diagnosis of thelaziosis and the identification/differentiation of species heavily relies on morphological features. Therefore, we conducted an epidemiological study in Romanian cattle, with the aim to obtain morphological and molecular data that can be used for species identification. METHODS: The eyes of 705 slaughtered cattle were sampled and subjected to morphological identification, morphometric analysis, and molecular characterization. PCR amplification and sequence analysis were performed based on the cytochromec oxidase subunit 1 (COI) gene. Statistical tests assessed the correlations between infection parameters and ecological or biogeographical factors. A novel PCR method was developed based on the consensus sequence from each species. Specific forward primers were designed for each of the three species, and a reverse primer (COIintR) was used for all reactions. A consensus thermal profile was established by gradient PCR amplification of each species separately. RESULTS: Of the sampled cattle, 19.3% were infected with Thelazia spp. Prevalence varied significantly with ecogeographical factors. A total of 585 Thelazia nematodes were recovered, with T. rhodesi being the most abundant, followed by T. skrjabini and T. gulosa. Morphometric and molecular analyses supported the morphological identification, yielding unique sequences for each species. From the 59 T. rhodesi specimens sequenced, 29 unique sequences were obtained, with a 99.1-99.85% nucleotide identity to the only other COI sequence present in GenBank®. All nine T. gulosa isolates were unique (99.37-100% nucleotide identity to other sequences), while T. skrjabini specimens displayed 98.47-100% nucleotide identity to the sole available sequence. CONCLUSIONS: Bovine thelaziosis is prevalent in Romania, raising concerns for animal welfare and potential economic impacts. Infected cattle grazing alongside vulnerable wild ruminants, such as the European bison, may affect conservation efforts. Our newly developed multiplex PCR shows promise as a valuable surveillance tool, enabling the detection of occult infections in apparently healthy animals through lachrymal secretion testing.

The European Badger as a New Host for Dirofilaria immitis and an Update on the Distribution of the Heartworm in Wild Carnivores from Romania.

Dirofilaria immitis is an important mosquito-borne nematode, being of both veterinary and public health concern. The typical final host is represented by the domestic dog, Canis familiaris, but it is able to infect a large variety of mammalian species. During the present study (March 2016-February 2022), a total of 459 wild carnivore carcasses belonging to 17 species, from Romania, were evaluated for the presence of adult D. immitis by necropsy. Overall, 20 animals (4.36%) were positive: twelve golden jackals, C. aureus (19.05%); four red foxes, Vulpes vulpes (6.67%); one raccoon dog, Nyctereutes procyonoides; two wild cats, Felis silvestris (4.65%); and one European badger, Meles meles (0.87%). This study provides further evidence of the occurrence of the canine heartworm, D. immitis, in Romania, expanding the known host spectrum, reports a new host species for this parasite, the European badger, and a new host for Europe, the raccoon dog.

New insights into the distribution of cardio-pulmonary nematodes in road-killed wild felids from Romania.

BACKGROUND: The population of wild felids is large and stable in Romania with many carnivore habitats being protected. Felids can be infected with a wide variety of cardio-pulmonary nematodes and can act as reservoirs of infection for domestic cats. The aim of this study was to evaluate the distribution and species diversity of cardio-pulmonary nematodes in wild felids from Romania. METHODS: A total of 54 wild felids (7 Lynx lynx and 47 Felis silvestris) were legally collected from different locations in Romania and analysed by complete parasitological necropsy. The entire respiratory tract was longitudinally opened and checked for the presence of nematodes. Detected nematodes were collected and morphologically identified to species level. RESULTS: Two Eurasian lynxes and 29 European wildcats were positive for cardio-pulmonary nematodes. Eurasian lynxes were infected with two species of cardio-pulmonary nematodes, Eucoleus aerophilus and Troglostrongylus brevior, while in wildcats the dominant parasite was E. aerophilus (34.0%) followed by Angiostrongylus chabaudi (23.4%) and T. brevior (14.9%). Dirofilaria immitis and Aelurostrongylus abstrusus were each detected in two wildcats (4.3%). CONCLUSIONS: The present study expanded the epidemiological knowledge on felid cardiopulmonary nematodes in Romania. We confirmed the presence of A. abstrusus in wildcats and a patent infection with T. brevior in Eurasian lynx.

A review of Trichinella species infection in wild animals in Romania.

Nematodes of the genus Trichinella are important zoonotic parasites present throughout Romania. This study aimed to assess the status of Trichinella species in wild animals in Romania over the past 30 years. A literature review of original studies concerning the only two species (out of the four in Europe) of Trichinella (T. spiralis and T. britovi) confirmed in wildlife from Romania was conducted and corroborated with the results of our original research concerning the topic. This review article has shown that, in Romania, European minks were infected with T. spiralis, while wolves, European wild cats, Eurasian lynx, golden jackals, stone marten, and European badgers were infected with T. britovi, respectively. Both Trichinella species have been identified in foxes, bears, wild boars, and ermines, but mixed infections have been found only in European polecats. Trichinella infection is still significantly present in Romania, infecting several wild omnivorous and carnivorous species in an equal manner, with different prevalence rates over the years. Regarding the spatial distribution of T. spiralis and T. britovi in Romania, both species can be found all over the country, but in wild animals, T. britovi is the most prevalent.

European Hares, Lepus europaeus, Represent a Reservoir Host for Thelazia callipaeda in Romania.

Thelaziosis caused by Thelazia callipaeda is an emerging disease in Europe. Only two reports of naturally infected lagomorphs have been published so far. The aim of this study was to evaluate the status of the Romanian populations of European brown hares, Lepus europaeus as reservoir hosts for T. callipaeda. Between November 2019 and November 2021, the eyes of 326 L. europaeus carcasses were examined for the presence of ocular parasites. Nematodes were stored in plastic vials with physiological saline, followed by morphological and molecular identification. QGis 3.20 and EpiInfoTM 7 were used for mapping and statistical analysis. Four (1.23%) hares harbored T. callipaeda infection, with a total of 84 nematodes collected (mean intensity 21 nematodes/host), with 45 males, 39 females (two sexually immature, seven with only eggs, and 30 with eggs and larvae). One specimen from each host was successfully sequenced resulting in a 100% similarity with several other sequences of T. callipaeda haplotype 1. Statistical analysis revealed no significant results. The current study represents a first report of T. callipaeda in the European brown hare in Romania, and the second in Europe, also reiterating the role of lagomorphs as reservoir hosts for this zoonotic ocular nematode.

Prevalence of Swine Gastrointestinal Parasites in Two Free-Range Farms from Nord-West Region of Romania.

Parasitic diseases cause significant economic losses in swine, including free-range swine farms, the number of which in Romania has increased in the last decades. The current study aimed to identify the parasitic profile of swine raised on two free-range (low-input) farms from Transylvania. Nine hundred sixty samples collected from weaners, fatteners, and sows were investigated by flotation, centrifugal sedimentation, modified Ziehl-Neelsen stained fecal smear, modified Blagg technique, and oocyst/egg cultures. The number of oocysts (OPG), cysts (CPG), and eggs (EPG) were counted per gram of fecal matter. The examination revealed parasitic infections with Balantidium coli, Eimeria spp., Ascaris suum, Trichuris suis, Oesophagostomum spp., Strongyloides ransomi and Cryptosporidium spp. Prevalence (P) and the mean intensity (MI) of the infections varied according to age, swine category, farm, and season. The overall prevalence in both free-range farms according to the age category was 63.2%­Eimeria spp., 70.31%­B. coli, 9.38%­Oesophagostomum spp., 3.75% S. ransomi, and 18.12% Cryptosporidium spp. in weaners. In fatteners Eimeria spp. revealed a prevalence of 50.93%, B. coli­72.5 %, A. suum­63.13%, T. suis­39.06%, and in sows Eimeria spp.­39.06%, B. coli­62.19%, A. suum­34.06%, Oesophagostomum spp.­27.19%, S. ransomi­1.56% and Cryptosporidium spp.­9.38%. The study revealed statistically significant (p < 0.05) differences between age groups, seasons, and farms for all diagnosed parasites. Further research is required to better understand the epidemiology of these infections in swine from Transylvania.

Antiparasitic Action of Lactobacillus casei ATCC 393 and Lactobacillus paracasei CNCM Strains in CD-1 Mice Experimentally Infected with Trichinella britovi.

Nematodes of the genus Trichinella are among the most widespread parasites of domestic and wild omnivores and predatory animals. The present study aimed to evaluate the antiparasitic effect of Lactobacillus casei ATCC 393 (original) and L. paracasei CNCM in CD-1 mice experimentally infected with Trichinella britovi. Four groups of 20 mice (10 females and 10 males/group) were used, with two control (C) groups and two experimental (E) groups, in which each animal received a daily oral dose of 100 µL of 105 CFU/mL probiotics in Ringer's solution. On day 7, all mice (except the negative control group) were infected orally with Trichinella (100 larvae/animal) as well as the two probiotics. On day 9 post-infection (p.i.), 10 mice/group were euthanized, and the presence of adult parasites in the intestinal content and wall was tested. On day 32 p.i., 10 mice/group were euthanized, then trichinoscopy and artificial digestion were performed to assess the muscle infection with T. britovi. On day 9 p.i., the experimental group pretreated with L. casei ATCC 393 (6.3 ± 3.03) showed a significantly lower number of adult parasites in the intestinal wall compared with the positive control group (24.6 ± 4.78). Additionally, a significantly lower adult parasite count in the intestinal wall was registered in female mice pretreated with L. paracasei CNCM (7.4 ± 4.71) compared to female mice from the positive control (29.0 ± 5.17). No statistically relevant results were obtained concerning the male mice or the data obtained at 32 days p.i., irrespective of mice gender.

Spotted Fever Group Rickettsia spp. Diversity in Ticks and the First Report of Rickettsia hoogstraalii in Romania.

Tickborne bacterial pathogens have been described worldwide as risk factors for both animal and human health. Spotted fevers caused by Rickettsiae may cause non-specific symptoms, which make clinical diagnosis difficult. The aim of the current study was to evaluate and review the diversity of SFG Rickettsiae in ticks collected in 41 counties in Romania. A total of 2028 questing and engorged ticks collected in Romania belonging to five species were tested by PCR amplification of Rickettsia spp. gltA and 17-D gene fragments: Ixodes ricinus (n = 1128), Dermacentor marginatus (n = 507), D. reticulatus (n = 165), Rhipicephalus rossicus (n = 128) and Haemaphysalis punctata (n = 100). Five Rickettsia species were identified following DNA sequence analysis: R. helvetica, R. monacensis, R. slovaca, R. raoultii, and R. hoogstraalii. The most common species detected was R. monacensis. Moreover, R. hoogstraalii was detected for the first time in Romania and in R. rossicus ticks. The detection of R. raoultii and R. monacensis in questing larvae of Hae. punctata suggests the possible transovarial transmission of these Rickettsia species in ticks. The detection of R. hoogstraalii for the first time in Romania increases the reported SFG Rickettsia diversity in the country.

Thelazia rhodesi in a dairy farm in Romania and successful treatment using eprinomectin.

Bovine thelaziosis can be caused by Thelazia rhodesi Desmarest 1828, Thelazia gulosa Railliet & Henry 1910 and Thelazia skrjabini Erschow 1928 which are known to be transmitted by species of Muscidae. Therapeutic measures in cattle include mechanical removal of the adult parasites, followed by irrigation of the conjunctival sac with different solutions such as levamisole (1%) or administration of ivermectin (0.2 mg/kg SC), doramectin (0.2 mg/kg SC or pour-on) both with high efficacy, however, the use of such macrocyclic lactones can be problematic in lactating cattle. Nematodes harboured in the eyes of some cattle from Romania were morphologically identified as Thelazia rhodesi and the PCR analysis confirmed the diagnosis with a 98.0% nucleotide similarity for ITS1 sequence, with other sequences available in GenBank. The present paper reports the presence of T. rhodesi in cattle from a dairy farm in Romania and the recovery after treatment with eprinomectin. To the best of our knowledge this is the first report on the efficacy of eprinomectin against Thelazia spp. and the first molecular confirmation of T. rhodesi in Romania.

Subcutaneous ticks: a first report in a golden jackal, and their absence in non-canid carnivores.

BACKGROUND: Ticks are hematophagous arthropods which normally attach to the surface of the host's skin. Their aberrant presence in the subcutaneous tissue of a few carnivores, predominantly foxes, has been reported. However, there have been no reports of this phenomenon in other carnivores such as mustelids or golden jackals. Our aim was to investigate the host spectrum for this aberrant localization of ticks. METHODS: Between 2015 and 2020, a total of 198 carcasses of 12 species of carnivore were examined by parasitological necropsy. When a subcutaneous tick was found, the nodule was removed, carefully dissected, and stored in ethanol. The morphological identification of the subcutaneous tick was carried out to species level. RESULTS: A single subcutaneous tick was found in one carcass, that of a golden jackal (Canis aureus). The tick was identified as a female Ixodes ricinus. All the other carcasses were negative for the presence of subcutaneous ticks. CONCLUSION: To our knowledge, this is the first report of a subcutaneous tick in a golden jackal. This finding broadens the host spectrum of subcutaneous ticks, and reinforces the idea that, among carnivores, this phenomenon only occurs in canids.

Reprint of: The European badger, Meles Meles, as a new host for Trichinella britovi in Romania.

The European badger, Meles meles (Carnivora, Mustelidae) is a widespread opportunistic omnivorous mammal. Its food spectrum comprises a wide variety of plants and small animals, occasionally including rodents, such as mice or rats. Considering that rodents are known to play a key role in the life cycle of Trichinella spp., the aims of this study were to investigate the occurrence of these parasites in badgers from Romania and to identify the species. Overall, 61 badgers originating from 14 counties were examined by trichinoscopy and artificial digestion. For species determination, the positive muscle samples, and the larvae recovered from the artificial digestion were used for DNA isolation, and further processed by multiplex PCR. A single badger, originating from Sibiu County, Central Romania, was found positive for Trichinella spp. Five cysts were identified using trichinoscopy: four in the diaphragm and one in the foreleg muscles. Artificial digestion revealed an infection rate of 70 larvae/100 g of muscle. The PCR indicated the occurrence of T. britovi, which is the most commonly detected species in wild carnivores in temperate areas. Although T. britovi has previously been reported in Romania, this represents the first report of its occurrence in the European badger in the country. However, the low prevalence indicates a minor reservoir role of this species.

Babesia pisicii n. sp. and Babesia canis Infect European Wild Cats, Felis silvestris, in Romania.

Haemoparasites of the genus Babesia infect a wide range of domestic and wild animals. Feline babesiosis is considered endemic in South Africa, while data on Babesia spp. infection in felids in Europe is scarce. Using samples from 51 wild felids, 44 Felis silvestris and 7 Lynx lynx, the study aimed to determine the presence and genetic diversity of Babesia spp. in wild felids in Romania by analyzing the 18S rDNA and two mitochondrial markers, cytochrome b (Cytb) and cytochrome c oxidase subunit I (COI) genes. By 18S rDNA analyses, Babesia spp. DNA was detected in 20 European wild felids. All sequences showed 100% similarity to B. canis by BLAST analysis. Conversely, Cytb and COI analyses revealed the presence of two Babesia spp., B. pisicii n. sp., which we herein describe, and B. canis. The pairwise comparison of both mitochondrial genes of B. pisicii n. sp. showed a genetic distance of at least 10.3% from the most closely related species, B. rossi. Phylogenetic analyses of Cytb and COI genes revealed that B. pisicii n. sp. is related to the so-called "large" canid-associated Babesia species forming a separate subclade in a sister position to B. rossi.