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Three species of the genus Angiostrongylus are known to infect European carnivores: A. vasorum (mainly in canids but also in other carnivores), A. chabaudi (in felids) and A. daskalovi (in mustelids). A. vasorum is responsible for clinically severe disease in domestic dogs, most commonly diagnosed based on fecal examination and serological detection of circulating antigens. Considering the poorly known host range and the challenging larval differentiation in the feces between the three species of Angiostrongylus infecting European carnivores, our aim was to evaluate the cross-reactivity of A. chabaudi and A. daskalovi with A. vasorum using a commercial serologic test developed for domestic dogs. Badgers (Meles meles) (n = 10) and wildcats (Felis silvestris) (n = 8) were examined between 2015 and 2016 by full parasitological necropsy with subsequent morphological and molecular identification of nematodes and by serology, using IDEXX Angio Detect™ tests. Five out of the ten badgers and two out of the eight wildcats were harboring nematodes in the pulmonary arteries. All nematodes were identified morphologically as A. daskalovi in badgers and A. chabaudi in wildcats, respectively. Serological examination of the plasma samples revealed the positivity of the same animals as found in necropsy. None of the animals negative at necropsy was positive at serology. The 100% correlation between the necropsy results and the serologic positivity to IDEXX Angio Detect™ in badgers infected with A. daskalovi and wildcats infected with A. chabaudi suggest that these rapid tests are able to identify circulating antigens of all species of Angiostrongylus found in European carnivores: A. vasorum, A. daskalovi and A. chabaudi. The possibility for future in-clinic use of this test in domestic cats should be further investigated.

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.

Background Small mammals play an important role in the life-cycle of ticks and are reservoirs for several zoonotic pathogens. The aim of this study was to provide epidemiological data regarding the presence of Borrelia spp. in tissues of small mammals from Romania. Methods We examined 401 individuals belonging to 11 small mammal species collected in Romania. Collections cover the largest effort to survey these reservoirs in the country. Tissue samples were analyzed by multiplex qPCR targeting the ospA gene of Borrelia burgdorferi (s.l.) and a part of the flaB gene of B. miyamotoi. Positive samples were further analysed by conventional PCR and sequenced. Results The overall prevalence of infection with Borrelia spp. in small mammal tissues was 4.9%. The most commonly detected species were B. afzelii, followed by B. garinii/B. bavariensis, B. miyamotoi and B. burgdorferi (s.s.). To our knowledge, we report for the first time the detection of Borrelia spp. in Crocidura leucodon and C. suaveolens, and B. miyamotoi in the liver of Myodes glareolus. Conclusions To our knowledge, our study evaluates for the first time the occurrence of Borrelia spp. in small mammals in Romania, contributing to a better knowledge of the distribution of these bacteria. This survey upgrades previous data on the spatial distribution of the pathogens and reveals the importance of animal surveillance regarding Lyme borreliosis and relapsing fever caused by B. miyamotoi.

Nematodes of genus Trichinella are wide-spread zoonotic parasites, able to infect a large variety of vertebrates. Animal hosts are usually regarded as asymptomatic carriers. However, there is little data regarding the functional consequences that T. spiralis infection renders on muscle cells. The aim of the present study was to assess the effect of T. spiralis on the effort capacity of experimentally infected mice. Overall, 60 mice, divided into three groups were used: M (uninfected), L200 and L1000, infected with 200 or 1000 larvae/mouse respectively. The mice were periodically weighed and their effort capacity was evaluated (days 0, 7, 15, 35 and 60). From each group, two randomly selected mice were euthanized after evaluation carcasses were artificially digested in order to establish the number of larvae per gram (LPG). On day 0, there were no significant differences among groups. Starting with day 7, the effort capacity of infected groups decreased, with significant differences between group M and the infected groups. From day 15, the differences between the infected groups also became significant. The LPG gradually increased and the differences between groups were always significant. A strong correlation between the LPG and decreased effort capacity was noted. The present study demonstrates the reduction of muscular capacity in mice experimentally infected with Trichinella spiralis, in correlation with the infective dose, providing new insights in this parasite's transmission strategy.

Background Thelazia callipaeda (Spirurida, Thelaziidae) is a vector-borne zoonotic eye worm with a broad host spectrum. In Europe, it is an emerging threat, having greatly expanded its geographical distribution during the past two decades. In Romania, T. callipaeda has been previously reported in domestic and wild canids and felids. The aim of the present study was to assess the occurrence of T. callipaeda in mustelids in the country. Methods Between March 2015 and April 2019, 77 road-killed mustelids (3 pine martens, Martes martes; 6 European polecats, Mustela putorius; 13 beech martens, Martes foina; and 55 European badgers, Meles meles) were examined by necropsy. If present, all ocular nematodes were collected and stored in absolute ethanol, for subsequent morphological and molecular identification. Results Two animals were found to be infected with T. callipaeda: one European badger and one beech marten. The molecular analysis revealed a 100% nucleotide similarity to T. callipaeda haplotype h1 for all the sequenced specimens. Conclusions To our knowledge, the present study demonstrates for the first time the occurrence of T. callipaeda in mustelids from Romania, records the easternmost locality of the parasite in Europe, and represents the first report of T. callipaeda in the European badger, Meles meles, extending the known host range for this parasite in Europe.

Protists of the genus Cytauxzoon infect a wide variety of wild and domestic felids worldwide. While the American Cytauxzoon felis has been well described, data on the European isolates of Cytauxzoon are still scant. The aim of the current study was to determine the genetic diversity of European Cytauxzoon spp. in wild felids across Europe by analyzing one nuclear and two mitochondrial genes, along with representative complete mitochondrial genomes. Overall, 106 biological samples from wild felids (92 from Felis silvestris and 14 from Lynx lynx) from Germany, Romania, Czech Republic, and Luxembourg were collected and screened for the presence of Cytauxzoon spp. using nested PCR protocols, targeting the highly conserved 18S rDNA, mitochondrial cytochrome b (CytB) and cytochrome c oxidase subunit I (COI) genes. Furthermore, 18 previously confirmed wild felid biological samples from Europe, and comparative material from USA positive for C. felis, were included in the study. In 18S rDNA sequences analyses, Cytauxzoon spp. from felids formed two separate clades of New World and Old World isolates, with a low inner diversity of the European clade. In contrast to 18S rDNA, the phylogenetic analyses of CytB and COI genes affirmatively revealed three highly supported clades, resulting in three defined genotypes. Similar intra- and interspecific variability of CytB and COI genes was observed in the case of different Babesia spp. Considering geography, host species and analyses of three genes, we conclude that the three detected genotypes of Cytauxzoon in European wild felids represent three new species, which we herein describe.

The risk of developing Lyme borreliosis (LB) after the bite of a Borrelia (B.) burgdorferi sensu lato (s.l.) infected tick in Romania is unknown.The present prospective study, performed in 2010-2011 in a hospital in Romania, has followed-up clinical and serological outcome of patients that presented with B. burgdorferi positive Ixodes (I.) ricinus bite. A second group of patients, including age, sex and residence-matched individuals bitten by B. burgdorferi negative ticks, was followed-up as a control group. The subjects' outcome was evaluated one year after the tick bite.Forty-three out of 389 ticks detached from patients were positive by hbb Real-Time PCR (RT-PCR) for B. burgdorferi s.l. (mainly B. afzelii, but also B. garinii, B. burgdorferi sensu stricto, B. spielmanii/B. valaisiana and B. lusitaniae). Twenty patients bitten by B. burgdorferi positive ticks and twenty matched control patients returned for the one year follow-up. Two patients from the B. burgdorferi positive group developed clinical manifestations of acute LB (erythema migrans) and 5 patients seroconverted (two from the B. burgdorferi positive group and three from the B. burgdorferi negative group). Borrelia afzelii was identified in ticks collected from persons that developed erythema migrans (EM). Comparing the two groups of patients, no statistical significant differences were found regarding presence of clinical symptoms or seroconversion.No outcome differences were found between the group of patients bitten by B. burgdorferi positive ticks and the group of patients bitten by B. burgdorferi negative ticks.

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.

Ixodes (Afrixodes) aulacodiArthur, 1956 is a poorly known species that has been recorded predominantly in the wet countries of western and central Africa, mainly associated to the greater cane rat Thryonomys swinderianus (Temmink). We herein redescribe the female, describe the male (ascribed to the species from specimens found in copula) and provide the 16S rDNA sequence. We also provide complete illustrations of the adults based on specimens found on greater cane rats in Ivory Coast. Ixodes aulacodi is included in the group of species of the subgenus Afrixodes that have horseshoe shaped anal groove, and which lack auriculae and cornua. The female is easily separated when compared with other species because of a unique combination of characters: All the coxae have internal spurs, coxa II has two external spurs, syncoxae are absent, and trochanters I-III have one spur each. The male has a notched hypostome and lacks syncoxae, auriculae and cornua.

The objective of this study was to evaluate different methods used for detection of Borrelia burgdorferi sensu lato (s.l.) in ticks: immunohistochemistry followed by focus floating microscopy (FFM) and real-time polymerase chain reaction (real-time PCR) targeting the ospA and hbb genes. Additionally, an optimized ospA real-time PCR assay was developed with an integrated internal amplification control (IAC) for the detection of inhibition in the PCR assay and was validated as an improved screening tool for B. burgdorferi. One hundred and thirty-six ticks collected from humans in a hospital from Cluj-Napoca, Romania, were investigated regarding genus, stage of development and sex, and then tested by all three assays. A poor quality of agreement was found between FFM and each of the two real-time PCR assays, as assessed by concordance analysis (Cohen's kappa), whereas the agreement between the two real-time PCR assays was moderate. The present study argues for a low sensitivity of FFM and underlines that discordant results of different assays used for detection of B. burgdorferi in ticks are frequent.

Cysticercosis is a zoonotic disease that may have a significant impact on the public health. The aim of the current study was to assess the seroprevalence of porcine cysticercosis in Northwestern Romania and the influence of several risk factors such as age, sex and season. The study included 1,288 pigs originating from 7 different counties and was conducted between May 2014 and May 2015. All pigs were reared in the traditional backyard pig system. Of these, 226 were young and 1,062 were adult pigs, 188 were males and 1,100 females. Serum samples were collected. The apDia Cysticercosis Antigen ELISA (apDia, Belgium) commercial kit was used to identify the viable cysticerci of Taenia sp. Out of the 1,288 serum samples, 83 were seropositive (seroprevalence: 6.4%). The risk factors’ analysis showed that age (3.1% juveniles vs 7.1% adults [P < 0.03]) and season (9.3% fall vs 11.8% winter [P P P = 0.1). Considering the genus specificity of ELISA commercial kit, the serology should be correlated with the post mortem examination in order to confirm the disease and have a Cysticercus cellulosae specific prevalence.

Background A century of debates on the taxonomy of members of the Metastrongyloidea Molin, 1861 led to many reclassifications. Considering the inconstant genus assignation and lack of genetic data, the main aim of this study was to support the validity of the genus Perostrongylus Schlegel, 1934, previously considered a synonym of Aelurostrongylus Cameron, 1927, based on new molecular phylogenetic data and to understand its evolutionary relationships with other metastrongyloid nematodes. Results Specimens of lungworm collected from European badgers in Germany, Romania and Bosnia and Herzegovina were morphologically and molecularly (rDNA, cox1) characterized. From a phylogenetic standpoint, Perostrongylus is grouped with high support together with the genera Filaroides van Beneden, 1858 and Parafilaroides Dougherty, 1946 and includes probably two species: Perostrongylus falciformis (Schlegel, 1933), a parasite of Meles meles in Europe and P. pridhami (Anderson, 1962), a parasite of Neovison vison in North America. Perostrongylus and Aelurostrongylus are assigned to different clades. Aelurostrongylus becomes a monotypic genus, with the only species Aelurostrongylus abstrusus (Railliet, 1898). In addition, we provide morphological and morphometric data for the first-stage (L1), second-stage (L2), and third-stage (L3) larvae of P. falciformis and describe their development in experimentally infected Cornu aspersum snails. The pathological and histopathological lesions in lungs of infected European badgers are also described. This is the first record of P. falciformis in Romania. Conclusions Molecular phylogenetic and morphological data support the validity of the genus Perostrongylus, most probably with two species, P. falciformis in European badgers and P. pridhami in minks in North America. The two genera clearly belong to two different clades: Perostrongylus is grouped together with the genera Filaroides and Parafilaroides (both in the family Filaroididae Schulz, 1951), whereas Aelurostrongylus belongs to a clade with no sister groups.

Background There is a need for experimental animal models for inflammatory bowel diseases (IBD), but no proposed model has been unanimously accepted. Objectives The aim of this study was to develop 2 affordable models of IBD in rats and to compare them. Material and methods We produced IBD in rats using either dextran sodium sulfate (DSS) or 2, 4, 6-trinitrobenzene sulfonic acid (TNBS). The requirements for experimental models were: a predictable clinical course, histopathology and inflammation similar to human ulcerative colitis (UC) and Crohn's disease (CD). The effect of acute administration of DSS and TNBS on oxidative stress (as measured by the assessment of glutathione peroxidase - GPx) was verified. The activity of whole blood GPx was measured using a commercially available Randox kit (Crumlin, UK). Results The administration of DSS increased GPx activity compared to the control and TNBS-treated groups, but not to a statistically significant degree. Histological examination of the colonic mucosa following the administration of DSS showed multifocal erosions with minimal to mild inflammatory infiltrate, mainly by polymorphonuclear cells (PMN), lymphocytes and plasma cells. For TNBS-induced colitis, the histological changes manifested as multifocal areas of ulcerative colitis with mild to severe inflammatory infiltrate. Whole blood GPx values displayed a direct dependence on the chemical agent used. Our results show a correlation between histopathology, proinflammatory state and oxidative stress. Conclusions The experimental DSSor TNBS-induced bowel inflammation used in this study corresponds to human IBD and is reproducible with characteristics indicative of acute inflammation in the case of the protocols mentioned.

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

Red foxes, Vulpes vulpes are among the most widely spread carnivores in the world, invading also urban areas and are often parasitized by various ticks and directly exposed to several vector-borne pathogens, including the commonly present “Theileria annae”. Considering the paucity of data on the possible vectors of this pathogen and the presence of the infection in various locations across the globe, the aim of our study was to understand the potential role of various environmental factors on the distribution of “T. annae” in red foxes from a well-defined region within the Carpathians, Romania. Between July 2016 and April 2017, a total of 347 blood samples originating from red foxes from 13 counties were tested using a PCR specifically designed for “T. annae”. In order to assess the potential distribution of “T. annae” based on niche modelling, we used presence-only data and 15 ecological variables. The probability of presence models was built using MaxEnt software. Of all sampled foxes, 20.1% (66 unique locations in 8 counties) were positive for “T. annae” DNA. There was no significant difference between the prevalence in males and females, nor between juveniles and adults. The sequences were all identical to each other and showed 100% identity to other sequences deposited in GenBank. The highest contribution to the spatial model was represented by the agricultural land coverage. This is the first study to demonstrate the presence of “T. annae” in foxes in Romania and the first spatial analysis for “T. annae” highlighting the importance of the environmental factors on its distribution.

Background Angiostrongylus vasorum is the causative agent of canine angiostrongylosis, a severe snail-borne disease of dogs. Red foxes are important natural reservoirs of infection, and surveys of foxes provide a more objective picture of the parasite distribution. Our aim was to investigate the possibility of the presence of A. vasorum in red foxes from the western part of Romania and to analyse the risk factors related to the sex, age and geographic origin of the foxes. Between July 2016 and April 2017, 567 hunted red foxes from 10 counties of western Romania were examined by necropsy for the presence of lungworms. Results Overall, the infection with A. vasorum has been found in 24 red foxes (4.2%) originating in four counties (Mureș, Hunedoara, Sălaj and Cluj). There was no significant difference between the prevalence in males and females, between juveniles and adults and between counties. Conclusions This is the first report of autochthonous infections of A. vasorum in Romania, showing a relatively low prevalence and extending eastwards the known distributional range of this parasite in Europe. The presence of autochthonous cases in domestic dogs in Romania remains to be confirmed by further studies.

We analyzed 32 specimens from nine species of Mustelidae for Trichinella; six infections from two Trichinella species were observed from three host species. This provides documentation of Trichinella in Mustela erminea and Martes foina in Romania and Trichinella spiralis in a mustelid host from Europe. Trichinella spiralis continues to be a public challenge characterized by a wide host range and geographical distribution ( Pozio 2007 ). During the past 20 yr, Romania has had the most reported human cases of trichinellosis in the world ( Blaga et al. 2007 ). Transmission occurs among domesticated swine, rats, and wild mammals that feed by scavenging or predation ( Pozio 2000 ). Trichinella transmission to humans may occur by consumption of meat of livestock infected after exposure to wildlife ( Pozio et al. 2009 ).

Brain samples from 182 red foxes (Vulpes vulpes) from Romania were examined using a standard PCR technique. Results provide evidence of Toxoplasma gondii (11 foxes = 6.0%) and Neospora caninum (1 fox = 0.5%) DNA in red foxes from Romania. No coinfections were found.

In this study, Hepatozoon species was molecularly identified and characterized for the first time on the Indian Ocean island of Mauritius. Partial sequences of the 18S rRNA gene of the Hepatozoon isolates were analysed from three naturally infected dogs. The sequences of H. canis were similar to the 18S rRNA partial sequences (JX112783, AB365071 99%) from dog blood samples from West Indies and Nigeria. Our sequences were deposited in the GenBank database.

The paper reports the prevalence and geographical distribution of Borrelia burgdorferi sensu lato (s.l.) and its genospecies in 12,221 questing Ixodes ricinus ticks collected at 183 locations from all the 41 counties of Romania. The unfed ticks were examined for the presence of B. burgdorferi s.l. by PCR targeting the intergenic spacer 5S-23S. Reverse line blot hybridization (RLB) and restriction fragment length polymorphism (RFLP) analysis were performed for identification of B. burgdorferi genospecies. The overall prevalence of infection was 1.4%, with an average local prevalence between 0.75% and 18.8%. B. burgdorferi s.l. was found in ticks of 55 of the 183 localities. The overall prevalence B. burgdorferi s.l. in ticks in the infected localities was 3.8%. The total infection prevalence was higher in female ticks than in other developmental stages. Three Borrelia genospecies were detected. The most widely distributed genospecies was B. afzelii, followed by B. garinii and B. burgdorferi sensu stricto (s.s.). The study is the first countrywide study and the first report of B. burgdorferi s.s. in Romania. The distribution maps show that higher prevalences were recorded in hilly areas, but Lyme borreliosis spirochetes were also present in forested lowlands, albeit with a lower prevalence.

The goal of this work was to identify novel, early antigens present in Trichinella spiralis. To this end, a cDNA library generated from 3-day old adult worms (Ad3) was immunologically screened using serum from a pig infected with 20,000 muscle larvae. The serum was obtained from multiple, time course bleeds coinciding with early worm development. Seventeen positive clones were isolated using serum obtained at 20 days post infection (dpi). All clones corresponded to one gene that exhibited high sequence identity with the T. spiralis ATP-dependent RNA helicase DDX19B which is involved in parasite growth and development. In addition, nine additional positive clones representing 5 unique genes were identified when the library was screened with 30 dpi serum; four of these five genes displayed high similarity with members of a putative T. spiralis serine protease family known to be involved in host invasion and host-parasite interactions. The remaining gene aligned with the T. spiralis hypothetical ORF 11.30. The identification of these antigens provides potential candidates for the early diagnosis of trichinellosis and for the development of a vaccine against this parasite.

Background Thelazia callipaeda is a vector-borne zoonotic nematode parasitizing the conjunctival sac of domestic and wild carnivores, rabbits and humans, with a vast distribution in Asia and the former Soviet Union. In Europe, the nematode has an emerging trend, being reported in Italy, France, Switzerland, Germany, Spain, Portugal, Bosnia and Herzegovina, Croatia, Romania, Greece and Serbia, with human cases known in Italy, France, Spain, Serbia and Croatia. In Romania, the infection was so far reported only in dogs, whereas there are no reports in wildlife despite the large numbers of wild carnivores in the country. The aim of this study was to evaluate the role of wild carnivores in the natural cycle of T. callipaeda in Romania. Methods Between 2014 and 2016, 89 wild carnivores (64 golden jackals, Canis aureus, 13 grey wolves, Canis lupus, nine wildcats, Felis silvestris and three Eurasian lynxes, Lynx lynx) have been examined. During the necropsy, both eyes of all the examined animals have been thoroughly inspected for the presence of parasites. If present, all nematodes were collected in absolute ethanol (for molecular analysis of the partial cox1 gene) or in 4 % formalin (for morphological identification). Results In total, three animals were found to be infected with T. callipaeda: a grey wolf, a golden jackal and a wildcat. The BLAST analysis of all the sequences showed a 100 % similarity to T. callipaeda haplotype h1. To the best of our knowledge, this study represents the first report of T. callipaeda in golden jackals, and the first study on T. callipaeda in wildlife from Romania. Conclusion Our data broaden the host spectrum and geographical distribution of T. callipaeda, highlighting the role of wild carnivores as natural reservoirs for the infection and confirming the ongoing expanding trend of this zoonotic nematode in Europe.

Anhemialges suteui n. sp. (Astigmata: Analgidae: Analginae) is described from the green hylia Hylia prasina (Cassin) (Passeriformes, Macrosphenidae) in Ivory Coast. The new species differs from the closest species, Anhemialges mironovi Kolarova, 2010, by the following features: in both sexes, solenidion σ on genu II is modified and widened in a form of leaf; dorsal setae c2, d2 and e2 are longer; and sternum and epimerites II are thinner. In males of the new species ambulacral discs of legs IV extend beyond the level of the lobar apices; in females, dorsal setae c2, d2 and e2 are situated on small oval shields and the anterior margin of epigynum exceeds the level of mesal extremities of epimerites II.

To the Editor: Ixodes spp. ticks are vectors for human and animal pathogens. Ix. ricinus ticks are widely distributed, frequently reported to feed on humans, and the main vector for a large variety of tickborne pathogens (1). The effect of ticks and tickborne diseases on public health, animal health and welfare, and animal production appears to be an increasing global problem, which will lead to considerable economic costs (2).

The aim of the present experiment was to assess the clinical, haematological, biochemical and immunological responses of pigs experimentally infected with Trichinella spiralis. One group of 6 pigs was infected with 1500 larvae/kg body weight and one group of 7, was maintained as control. The highest larval density was in the diaphragm (504.3 ± 8.2). During the experimental T. spiralis infection in pigs, increased values of total granulocyte number, eosinophils, blood glucose, K+ concentration, CK, ALT, AST and ALP were registered in different stages of infection. A slight increase of the phagocytic activity was registered 14 days p.i., in the infected group. Significantly increased values in lymphocyte activity, in infected group, was observed 30 and 60 days p.i. comparative with control. The specific IgG antibodies were detectable by ELISA at 14 days p.i. The results obtained in this study provide a better understanding about complete interactions occurring during Trichinella infection in pigs.

In 2010 and 2011, questing ticks were collected from 188 forested locations in all the 41 counties of Romania using the dragging method. The total of 13,771 ticks collected belonged to eleven species: Ixodes ricinus (86.9 %), Dermacentor marginatus (9.5 %), Haemaphysalis punctata (2.6 %), H. concinna (0.6 %), H. sulcata (0.3 %), H. parva (0.1 %), Hyalomma marginatum (0.02 %), D. reticulatus (0.02 %), I. crenulatus (0.007 %), I. hexagonus (0.007 %) and I. laguri (0.007 %). Ixodes ricinus was present in 97.7 % (n = 180) of locations, occurring exclusively in 41.7 % of the locations, whereas it was the dominant species in 38.8 % of the other locations, accounting for over 70 % of the total tick community. The following most common questing ticks were D. marginatus, H. punctata and H. concinna. Ixodes ricinus co-occurred with one, two or three sympatric species. The occurrence of D. reticulatus in forested habitats from Romania was found to be accidental.

Ticks and tick-borne diseases represent a great concern worldwide. Despite this, in Romania the studies regarding this subject has just started, and the interest of medical personnel, researchers, and citizens is increasing. Because the geographical range of many tick-borne diseases started to extend as consequences of different biological and environmental factors, it is important to study the diversity of ticks species, especially correlated with host associations. A total number of 840 ticks were collected between 1 April and 1 November 2010, from 66 animals, from 17 species in 11 counties, spread all over Romania. Four Ixodidae species were identified: Dermacentor marginatus (49.2%), Ixodes ricinus (48.3%), Hyalomma marginatum (2.4%), and Rhipicephalus sanguineus (0.1%). The obtained results indicate that D. marginatus is the most abundant tick species and I. ricinus is the most prevalent. As both of them are important vectors for human and animal diseases, the present paper discusses the associated risks for tick-borne diseases.

Previous studies in southeastern Europe reported a high incidence of human trichinellosis and a high prevalence of Trichinella infection in animals in countries like Bulgaria, Croatia, Romania and Serbia. The aim of this study was, using routine Trichinella test data in pig and game animals, to investigate the extent of Trichinella infection in slaughtered animals in Romania, over the period of 1997–2004, and to identify possible differences in prevalence among the various regions of Romania. Trichinella infection data were obtained from trichinelloscopic examinations of domestic (backyard and industrial reared pigs) and game species (wild boar and bears). A marked difference between Transylvania and other counties of Romania, observed for human trichinellosis, was taken into account when analyzing Trichinella epidemiological data. A cumulative prevalence of 8 cases/10 4 animals tested for pigs, 9 cases/10 3 animals tested for wild boars, and 13.1 cases/10 2 animals tested for bears was obtained for the 8 years period. Analysis of animal prevalence data demonstrated a geographical heterogeneity: whereas Trichinella prevalence in pigs is much lower in Transylvania than in the other counties, Trichinella prevalence in game animals is similar for the different regions. This observation suggests that, in Romania, rather than the levels of the parasite circulating in domestic and game animals, it was changes in the social and political structure of Romania in the 1990s, combined with inadequate meat inspection practices that were the main contributors to these geographic variations.

Trichinella spp. is a widely distributed nematode species affecting a large variety of hosts. Two major foci are identified in the natural cycle of the parasite: synanthropic and sylvatic. The sylvatic cycle includes two groups of animals: carnivores and omnivores. The carnivorous cycle of Trichinella involves large and small species as the bear, wolf, golden jackal, wildcat, lynx and mustelids, many of them common in Romania. The present study aimed to update the data on the prevalence of Trichinella spp. infection in large carnivores from Romania between 2014 and 2015 using the artificial digestion method. Out of 67 examined cadavers, 54 were golden jackals, 5 wolves, 5 wildcats and 3 Eurasian lynxes. All species were infected with Trichinella spp. and the prevalence was 40.0% in wolf, 33.3% in the wildcat, 53.7% in golden jackal and 66.6% in Eurasian lynx. These data demonstrate that wild carnivores remain important reservoirs for Trichinella in nature in Romania.

Background Anaplasma platys is a bacterium parasitic in the canine platelets, representing the causative agent of canine cyclic thrombocytopenia, with a worldwide distribution, mainly in tropical countries. The agent has zoonotic potential, being reported in several human clinical cases. The suspected vector is the tick Rhipicephalus sanguineus (sensu lato), widely distributed in animals and humans in the tropical regions of South America, Africa, Asia and Australia, but also in southern Europe. Only few scattered data concerning the epidemiology of A. platys in sub-Saharan Africa are available. The aim of our study was to evaluate the epidemiological status of A. platys in dogs and cats from rural communities in eastern and western Africa, where dogs and their ticks live in close contact with humans. Methods A total of 216 canine and 22 feline blood samples and ticks feeding on them were collected in 2013 and 2014 from eight localities in Ivory Coast and four localities in Kenya. PCR was performed using specific primers targeting a portion of the 16S rRNA gene, followed by sequencing. Results The main results highlight the high prevalence of A. platys infection in dogs in both Eastern and Western Africa and report it for the first time in Eastern Africa and in Rhipicephalus camicasi. Conclusions The presence of A. platys DNA in R. camicasi feeding on negative hosts together with the relatively high prevalence of A. platys, correlated with the absence of the probable vector Rhipicephalus sanguineus (s.l.) in Kenyan Island suggest the involvement of other tick species in the transmission of A. platys. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1443-3) contains supplementary material, which is available to authorized users.

Ixodes (Afrixodes) aulacodiArthur, 1956 is a poorly known species that has been recorded predominantly in the wet countries of western and central Africa, mainly associated to the greater cane rat Thryonomys swinderianus (Temmink). We herein redescribe the female, describe the male (ascribed to the species from specimens found in copula) and provide the 16S rDNA sequence. We also provide complete illustrations of the adults based on specimens found on greater cane rats in Ivory Coast. Ixodes aulacodi is included in the group of species of the subgenus Afrixodes that have horseshoe shaped anal groove, and which lack auriculae and cornua. The female is easily separated when compared with other species because of a unique combination of characters: All the coxae have internal spurs, coxa II has two external spurs, syncoxae are absent, and trochanters I-III have one spur each. The male has a notched hypostome and lacks syncoxae, auriculae and cornua.

Background Red foxes (Vulpes vulpes) are one of the most widespread wild carnivores in the world, being recognized to harbor and transmit a wide range of vector-borne diseases. Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato are zoonotic tick-borne pathogens causing emerging diseases. Wild animals play an essential role in the transmission of diseases and pathogens maintenance in nature. Epidemiological studies regarding the prevalence of tick-borne pathogens in red foxes are of public health importance, as they may successfully act as a pathogen transmission interface between wildlife, domestic animals and humans. Findings This study included 14 counties from Romania. A total number of 353 red foxes (Vulpes vulpes) were examined. Heart tissue samples were collected during necropsy and stored at −20 °C. Genomic DNA extraction was performed and all samples were examined by polymerase chain reaction (PCR). Specific primers for A. phagocytophilum, A. platys, E. canis and Borrelia burgdorferi s.l. were used. Sequence analysis was performed (Macrogen Europe, Amsterdam) and obtained sequences are available at GenBank™. Out of the 353 samples, 9 (2.55 %; 95 % CI: 1.25–4.96 %) were positive for A. phagocytophilum. Positive animals originated from 5 counties. In total, 5 out of 353 heart tissue samples (1.42 %; 95 % CI: 0.52–3.47 %) collected from red foxes were positive for B. burgdorferi s.l. Red foxes originated from 4 counties. None of the samples were positive for A. platys or E. canis. No co-infection with A. phagocytophilum and B. burgdorferi s.l. was found. Conclusion This first report of A. phagocytophilum and B. burgdorferi s.l. in red foxes from Romania suggests a limited role of foxes in the maintenance of the two related pathogens, but may represent a potential risk from a public health perspective.

Among wild carnivores, red foxes ( Vulpes vulpes ) are one of the most adapted to anthropic ecosystems and one of the most abundant ones living in close proximity to humans. They are well recognized as important natural reservoirs for a large number of zoonotic agents in Europe, including ticks and tick-borne pathogens. The aim of our study was to establish the prevalence of two zoonotic tick-borne pathogens, Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato (s.l.), in Ixodes ricinus ticks collected from red foxes. A total number of 216 red foxes, from 9 counties,were investigated and 204 Ixodes ricinus ticks, of all developmental stages were collected. PCR was performed on all samples to detect B. burgdorferi s.l. and A. phagocytophilum DNA. Only A. phagocytophilum was detected with an overall prevalence of 4.4%. As foxes represent a good model of sentinel species, further studies on the structure of tick communities correlated with the presence of tick-borne pathogens, can be suggestive for the risk posed for public health.

Granulocytic anaplasmosis is a common vector-borne disease of humans and animals with natural transmission cycle that involves tick vectors, among which Ixodes ricinus is the most important. The present paper reports the prevalence and geographical distribution of A. phagocytophilum in 10,438 questing Ixodes ricinus ticks collected at 113 locations from 40 counties of Romania. The unfed ticks were examined for the presence of A. phagocytophilum by PCR targeting a portion of ankA gene. The overall prevalence of infection was 3.42%, with local prevalences ranging between 0.29% and 22.45%, with an average prevalence of 5.39% in the infected localities. The infection with A. phagocytophilum was detected in 72 out of 113 localities and in 34 out of 40 counties. The highest prevalence was recorded in females followed by males and nymphs. The results and the distribution model have shown a large distribution of A. phagocytophilum, covering Romania's entire territory. This study is the first large scale survey of the presence of A. phagocytophilum in questing I. ricinus ticks from Romania.

Among wildlife, the red foxes (Vulpes vulpes) are one of the most adapted wild species to anthropic ecosystems. They are well recognized as important reservoirs for a large number of zoonotic agents in Europe, including ticks and tick-borne pathogens. Currently, there are few available data on the importance of red foxes in the ecoepidemiology of vector-borne diseases. Therefore, the aim of this study was to screen the dynamics of tick infestation in 357 red foxes from 12 Romanian counties. Tick identification was performed using the morphological keys. The overall prevalence of tick infestation was 43.7%. The 5753 collected ticks belonged to five species: Ixodes hexagonus (on 113 out of 156 foxes; prevalence 72.44%), I. ricinus (28.84%), I. crenulatus (7.7%), Dermacentor marginatus (7.05%) and Haemaphysalis punctata (0.64%). Coinfestation occurred in 24 foxes (22 with 2 tick species; 2 with 3 tick species) with the following associations: I. ricinus + I. hexagonus (n = 10), I. hexagonus + D. marginatus (n = 5), I. ricinus + I. crenulatus (n = 4), I. ricinus + D. marginatus (n = 2), I. hexagonus + I. crenulatus (n = 1), D. marginatus + I. hexagonus + I. ricinus (n = 1), and H. punctata + I. hexagonus + I. ricinus (n = 1). This study indicates that foxes are hosts to a relevant number of tick species with recognized vectorial role. The provided information can facilitate the understanding of the ecology of ticks and can be the basis for studies on the epidemiology of tick-borne diseases. The high prevalence of tick infestation in red foxes, coupled with the increasing presence of this species in synanthropic environments and a more nature-oriented lifestyle of people, can pose a significant threat to human health, through the emergence of tick-borne diseases. Moreover, foxes represent a good model of sentinel species. This research was performed as part of project IDEI PCE 236/2011.

Migratory birds play important roles as distributors of ticks within and between continents. In the Old World, the most important migratory route of birds links Asia, Europe and Africa. During their migration, birds use various stopover sites, where they feed and rest and where ticks may attach or detach, creating new natural foci for vector-borne diseases. Danube Delta is one of the most important migration hotspots and so far no studies were focused on ticks of migratory birds herein. The aim of the present study was to assess the species diversity and seasonal dynamics of ticks parasitizing migratory birds in Danube Delta Biosphere Reserve. Migratory birds were trapped on Grindul Lupilor (44°41′N; 28°56′E) using mist nets during 4 migratory seasons (2 spring and 2 autumn) in 2011 and 2012. From each bird, all the ticks were collected and identified based on morphological features. Epidemiological parameters (prevalence, mean abundance, mean intensity) were calculated and all data were analysed statistically based on the season (spring and autumn), regional status of birds (migrants and breeding) and foraging behaviour (ground feeders, reed-bed feeders, foliage feeders). A total of 1434 birds (46 species) were captured. Ticks were found on 94 birds (10 species). Significantly more migratory birds hosted ticks, compared to resident birds. The 400 collected ticks belonged to four species: Ixodes ricinus (92.25%), I. arboricola (6.25%), I. redikorzevi (1.00%) and Haemaphysalis punctata (0.50%). A higher prevalence was found for I. ricinus in spring, with higher prevalence of nymphs in this season, while larvae occurred with the same prevalence in both seasons. Larval intensity was higher during spring and nymphs were more abundant during autumn. The seasonal differences in our study may be related not to the local seasonal dynamics of ticks, but on the seasonal dynamics at the site of migration initiation.

Eucoleus aerophilus (syn. Capillaria aerophila) is a trichuroid nematode affecting domestic and wild carnivores and, sometimes, humans. This parasite has a worldwide distribution and may cause significant clinical disease in pet animals. The present paper investigates the sequence variation in partial mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of E. aerophilus isolates from pets and wild animals from different countries. Forty-four egg pools of E. aerophilus were collected from dogs, cats and foxes from Italy, while seventeen adult stages of E. aerophilus were obtained from red foxes and beech martens from Portugal, Romania, Serbia and UK. Fifteen different haplotypes were characterized and five were shared between pets in Italy and wildlife from Europe. The remaining haplotypes were either confined only in hosts or countries, or in a given host from a country. The phylogenetic analysis showed that all haplotypes clustered as a monophyletic group with a strong nodal support, indicating that all sequence types represented E. aerophilus. The results here presented have implications for a better understanding of the epidemiology, phylo-geography and clinical impact of E. aerophilus. In particular, the geographic distribution of E. aerophilus haplotypes in different host species and geographic regions, and their variation in terms of pathogenic impact and zoonotic role, warrant further investigations.

Background Hyalomma aegyptium is a hard-tick with a typical three-host life cycle. The main hosts are Palearctic tortoises of genus Testudo. However, other hosts can be used by immature ticks for feeding in natural conditions. Given this complex ecology and multiple host use, the circulation of pathogens by H. aegyptium between various hosts can be important from epidemiological point of view. The aim of this study was to evaluate the role of H. aegyptium as natural carrier of four important zoonotic pathogens. Methods From 2008 to 2011, 448 H. aegyptium ticks were collected from 45 Spur-thighed tortoises, Testudo graeca in Romania. DNA was extracted individually from each tick using a commercial kit. DNA was examined for the presence of specific sequences of Borrelia burgdorferi s.l., Anaplasma phagocytophilum, Ehrlichia canis and Coxiella burnetii by PCR, according to previously described protocols. Results PCR analysis of H. aegyptium revealed the presence of A. phagocytophilum (18.8%), E. canis (14.1%) and C. burnetii (10%). 32.4% of the ticks were infected with at least one pathogen and 9.8% had co-infections. The stages most frequently infected were nymphs (50%) followed by males (33.9%) and females (27%). The number of tortoises which harboured infected ticks was 27/45 examined (60%). From all tested T. graeca, 40% harboured ticks infected with A. phagocytophilum, 46.7% had ticks infected with E. canis and 33.3% had ticks with C. burnetii. This study reports for the first time the presence of A. phagocytophilum and E. canis in H. aegyptium. Conclusions The presence and relatively high prevalence of three important zoonotic pathogens in H. aegyptium raises the question of their epidemiologic importance in disease ecology. As tortoises are unlikely to be reservoir hosts for A. phagocytophilum and E. canis and both these pathogens are common in H. aegyptium, this is an important indication for (1) a possible increased host-switching behaviour of these ticks to competent reservoir hosts (i.e. hedgehogs) and (2) transstadial transmission. Furthermore, if we consider also the presence of C. burnetii, we conclude that T. graeca and its ticks should be evaluated more seriously when assessing the eco-epidemiology of zoonotic diseases.

Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are two tick-borne pathogens of medical and/or veterinary importance which are distributed worldwide. Erinaceus roumanicus , the Northern white-breasted hedgehog, is a common synanthropic species that is known to carry not only the hedgehog tick, Ixodes hexagonus , but also I. ricinus , the most common European tick species. I. ricinus is the main vector of both mentioned pathogens. Within this framework and because only limited information is available on the role of E. roumanicus in the ecology of B. burgdorferi s.l. and Anaplasma phagocytophilum in Europe, we carried out an epidemiological surveillance on this species in Romania. From the 57 examined hedgehogs collected in 12 different counties, 24 presented tick infestation. Most ticks ( n = 959) were morphologically identified as larvae, nymphs, or adults of I. ricinus . The prevalence of B. burgdorferi s.l. was 0.4%, and that of A. phagocytophilum 12%. In all positive cases for B. burgdorferi s.l., restriction fragment length polymorphism revealed the genospecies B. afzelii . In Romania, only limited information is available on the epidemiology of B. burgdorferi s.l. and A. phagocytophilum . As hedgehogs commonly share the same environment with humans and other potential reservoir hosts for tick-borne pathogens, our study provides new epidemiological data of public health importance.

Background Lyme disease is a widespread cosmopolitan zoonosis caused by species belonging to the genus Borrelia. It is transmitted from animal reservoir hosts to humans through hard - ticks of genus Ixodes which are vectors of the disease. Case presentation Borrelia burgdorferi sensu lato infection was identified in a marbled polecat, Vormela peregusna, and two European minks, Mustela lutreola, from Romania, by PCR. RFLP revealed the presence of a single genospecies, Borrelia burgdorferi sensu stricto. Conclusions This is the first report of the Lyme disease spirochetes in the two mentioned hosts.

The current paper is a synoptic review of the distribution and host associations of the 25 species of hard tick fauna (family Ixodidae) in Romania. In addition to a full literature survey, original data is presented, based on eight years of occasional or targeted sample collection. The literature data on geographical distribution was transposed digitally to the decimal degree coordinate system. For each species, an updated distribution map is given together with all historical data and new host associations. Overall, our paper records 58 new tick-host associations for Romania: 20 for Ixodes ricinus, 1 for I. apronophorus, 6 for I. arboricola, 2 for I. hexagonus, 9 for I. redikorzevi, 1 for I. trianguliceps, 2 for I. vespertilionis, 2 for Haemaphysalis punctata, 1 for H. sulcata, 2 for H. concinna, 1 for D. marginatus, 4 for Rhipicephalus sanguineus sensu lato, 1 for R. bursa and 6 for Hyalomma marginatum.

The overwhelming majority of animal conservation projects are focused on vertebrates, despite most of the species on Earth being invertebrates. Estimates state that about half of all named species of invertebrates are parasitic in at least one stage of their development. The dilemma of viewing parasites as biodiversity or pest has been discussed by several authors. However, ticks were omitted. The latest taxonomic synopses of non-fossil Ixodidae consider valid 700 species. Though, how many of them are still extant is almost impossible to tell, as many of them are known only from type specimens in museums and were never collected since their original description. Moreover, many hosts are endangered and as part of conservation efforts of threatened vertebrates, a common practice is the removal of, and treatment for external parasites, with devastating impact on tick populations. There are several known cases when the host became extinct with subsequent coextinction of their ectoparasites. For our synoptic approach we have used the IUCN status of the host in order to evaluate the status of specifically associated hard-ticks. As a result, we propose a number of 63 coendangered and one extinct hard-tick species. On the other side of the coin, the most important issue regarding tick-host associations is vectorial transmission of microbial pathogens (i.e. viruses, bacteria, protozoans). Tick-borne diseases of threatened vertebrates are sometimes fatal to their hosts. Mortality associated with pathogens acquired from ticks has been documented in several cases, mostly after translocations. Are ticks a real threat to their coendangered host and should they be eliminated? Up to date, there are no reliable proofs that ticks listed by us as coendangered are competent vectors for pathogens of endangered animals.

Herbivorous animals are usually, by virtue of their diet, outside the major transmission cycles of Trichinella spp. However, since 1975, the year of the first report of human trichinellosis caused by the consumption of infected horse meat, the domestic horse has appeared as a novel vector of Trichinella spp. infection to humans, with 15 outbreaks documented in France and Italy. Romania, one of the main countries exporting horses into the European Union (EU), experienced a dramatic increase of Trichinella spp. infection in both domestic pigs and humans in the 1990s. Some Trichinella spiralis-infected horses were exported to the EU during this period. The aim of this study was to evaluate the prevalence of Trichinella spp. infections in horses from Romania using both direct and indirect tests. Of 3000 serum samples tested in 2001, none were positive by ELISA using three different Trichinella antigens (crude; excretory/secretory, ES; stg-BSA antigens). Of 2992 serum samples tested in 2002, 17 (0.56%) showed optical density values higher than the cut-off in an ELISA using ES antigens and one was confirmed by western blot (WB). Four of the 17 ELISA positive horses, including the horse with a confirmed serology by WB, were subjected for intensive meat examination at slaughter, but no Trichinella spp. larvae were detected. Further, no Trichinella spp. larvae were detected by trichinelloscopy and artificial digestion of 25,838 horses slaughtered in Alexandria and Timisoara between 2001 and 2004. The false positive results obtained by serology confirm the previous work on the unreliability of serology for detection of Trichinella spp. infection in horses. Furthermore, the lack of detection of Trichinella spp. infected horses by artificial digestion, suggests a very low prevalence of infection in horses in Romania.

In order to evaluate prevalence and intensity of apicomplexan hemoparasites in free-ranging reptiles from Romania, blood smears were collected from European pond turtles (Emys orbicularis), sand lizards (Lacerta agilis), and spur-thighed tortoises (Testudo graeca). All three host species were positive for blood parasites, with prevalence of infected individuals between 60.71% and 100% and variable intensity. Similarities and differences with other epidemiological data are discussed.

Wild birds are hosts for several species of ticks, contributing to the maintenance of their local populations in delimited geographic areas. migratory birds play important roles as distributors of ticks within and between distant territories, including continents. Ticks collected from birds are responsible for hosting a significant number of human pathogens. The extensive wetland complex of the Danube Delta provides an internationally important stopover site for millions of birds, belonging to 300 different species, travelling annually. The aim of this study was to detect Anaplasma phagocytophilum in feeding ticks collected from migratory birds along four migratory seasons. Ticks were collected from the birds with forceps and preserved in 96% ethanol for later examination using a separate vial for each bird. A total of 1436 birds in 56 species (15 families) of Passeriformes and 3 non-Passeriformes bird species were captured. A total of 400 ticks were collected and identified as larvae (n = 191; 47.75%), nymphs (n = 201; 50.25%) or adult females (n = 8; 2%). No adult males were found. The ticks belonged to four species (Ixodes ricinus, I. arboricola, I. redikorzevi and Haemaphysalis punctata). Ixodes ricinus was the most common tick (369/400 ticks, 92.25% of the total collected), with a total of 181 larvae, 180 nymphs and 8 females. For detection of Anaplasma phagocytophilum all 400 ticks were examined by PCR, targeting the msp2 gene. Anaplasma phagocytophilum specific DNA was detected in 2 larvae and 2 nymphs of, I. ricinus, from 3 birds prevalence 1.08-%. The infection was absent from the other species of collected ticks The bird species that carried ticks infected with A. phagocytophilum were Turdus merula, Erithacus rubecula and Fringilla coelebs. Several other studies have demonstrated the presence of A. phagocytophilum in ticks collected from birds. Comparing our data with these results, we confirm the low prevalence of A. phagocytophilum, showingthat migratory birds, despite being without reservoir competencemight be important hosts for the dispersal of infected ticks.

Necropsy of 82 specimens of reptiles (snakes, lizards, and turtles) was performed between 2002 and 2006 to determine the helminth fauna. We examined 7 species of reptiles: Natrix natrix (n=25), Natrix tessellata (n=20), Vipera berus (n=5), Coluber caspius (n=12), Anguis fragilis (n=2), Lacerta agilis (n=10), and Emys orbicularis (n=8). Overall prevalence of helminthic infections in Romanian reptiles was 81.71%. In snakes, the most prevalent group of helminths was trematodes. The helminth species, their prevalence, intensity, and abundance are shown in Table 1. In two reptile species, all individuals were infected with helminths (N. natrix and E. orbicularis). The lowest prevalence of helminthic infection was in C. caspius. Three digenean trematodes, namely, Leptophallus nigrovenosus, Metaleptophallus gracillimus, and Spirhapalum polesianum, and three nematodes, Entomelas entomelas, Strongyloides mirzai, and Hexametra quadricornis, are new records for Romania. N. natrix has a new host record for infection with nematode genus Eustrongylides.

Background Dirofilaria immitis and Dirofilaria repens are mosquito-transmitted zoonotic nematodes, causing heartworm disease and skin lesions, respectively, in carnivores. In Europe, the domestic dog is apparently the main definitive host, but patent infections occur also in other species of carnivores. The rapid spread of the golden jackals (Canis aureus) throughout Europe opens a question of involvement of this species in the sylvatic cycle of pathogens in the colonised territories, including Dirofilaria spp. Methods Between January 2014 and May 2015, 54 golden jackals from 18 localities in Romania were examined by full necropsy for the presence of adult filarioid nematodes and blood samples from all animals were screened for the presence of microfilariae of D. immitis, D. repens and Acanthocheilonema reconditum by multiplex PCR DNA amplification. Results Nematodes morphologically identified as D. immitis were found in 18.52 % of the animals, originating from the southern part of Romania. No D. repens or A. reconditum were found at necropsy. The molecular prevalence in blood samples from the same animals was 9.26 % for D. immitis and 1.85 % for D. repens. All samples were negative by PCR for A. reconditum. Conclusion The relatively high prevalence of Dirofilaria spp. infections in golden jackals from Romania together with the increasing density of the jackal populations highlight their potential role in the transmission of these zoonotic parasites and in the maintenance of natural disease foci.

Larvae of Trichinella sp. were identified in a golden jackal (Canis aureus) from Romania by both trichinelloscopy and artificial digestion. The larvae were identified as Trichinella britovi using a multiplex polymerase chain reaction biotyping method. This is the first report of Trichinella sp. in a jackal in Romania.

International audience; According to the International Commission on Trichinellosis survey in 2004, Romania has the most cases of trichinellosis in the world. Epidemiologic data for each county were collected and analyzed from two different time periods: before (1980–1989) and after (1990–2004) political changes. Data were analyzed separately for Transylvania and the rest of the Romanian counties. During the past 25 years, 28,293 human cases of trichinellosis were reported with an incidence of 51.0 cases per 106 persons per year. An important increase in the incidence was observed from 1980 to 1989 compared with the 1990–2004 period. For the entire period, the incidence rate obtained for Transylvanian counties (82.2 cases per 106 persons per year) was higher than the incidence rate obtained for the other counties (35.7 cases per 106 persons per year). Hypotheses and facts contributing to the heterogeneity of human trichinellosis cases are discussed.