A proposed ectochory of Galba truncatula snails between wallow sites enhances transmission of Fascioloides magna at gemenc, in Hungary.

Each year temporary puddles are formed on compacted earth roads as a consequence of heavy rains and subsequent flooding in the large floodplain forests of the Danube River, Hungary, Central Europe, Hungary. After the receding of floodwaters, the muddy puddles persist from spring to mid-fall, where they are densely populated by Galba truncatula snails on an annual basis. These snails are the sole intermediate host of Fascioloides magna liver fluke of deer in the forest. According to field observations, G. truncatula is a very rare species on banks of river branches and lakes within the forest but always appears in large aggregations in these semi-natural potholes. Red deer (Cervus elaphus) leave their droppings in puddles while they wallow in the mud, therefore the snails encounter the miracidia of F. magna frequently. Snails are not able to dig themselves into the tamp soil of roads therefore they are destroyed by the wheels of vehicles which drive down the roads from autumn to the end of winter. Therefore, snails colonize these puddles regularly every spring. Since there is no connection between the distant puddles, it is supposed that deer and wild boars repeatedly introduce the snails into the puddles each year carrying snails with the help of mud, which is stuck on their fur while they wallow. This method of transport is supported by the fact that shell remnants of snails can be found on the bark of rubbing trunks situated nearby wallows. It seems that the sequential creation of wallow sites and the repeated introduction of G. truncatula enhance the likelihood for the trematode to infect both hosts.

Threat of cercarial dermatitis in Hungary: A first report of Trichobilharzia franki from the mallard (Anas platyrhynchos) and European ear snail (Radix auricularia) using molecular methods.

Cercarial dermatitis in humans is a re-emerging zoonotic disease caused by infectious larvae of avian blood flukes within the Schistosomatidae family. Upon water contact, these avian schistosome larvae directly penetrate human skin and cause irritation. Between September 2018 and September 2020, carcasses of 94 mallards (Anas platyrhynchos), two green-winged teals (Anas crecca) along with one ferruginous duck (Aythya nyroca) were examined. Birds were collected within 12 regions of Hungary, representative of 9 different counties. Inspecting both morphological characters and molecular data, the cytochrome oxidase subunit I (COI) and large subunit ribosomal DNA (LSU rDNA) sequences, Bilharziella polonica and Dendritobilharzia pulverulenta were each identified. Most importantly, Trichobilharzia franki was found for the first time in Hungary and in 5 dispersed counties in conjunction with of 3 counties where other avian schistosomes were found. Of note, these avian blood flukes were predominately encountered in wild mallards (∼50%) and not in birds reared for hunting (∼1%). In total, 245 European ear snails (Radix auricularia), a known intermediate host of Trichobilharzia spp., were collected from an urban pond in Eger, Hungary. Five snails (∼2%) consistently shed numerous furcocercariae of T. franki that were confirmed by molecular methods. Our findings help to pinpoint a contemporary life cycle of this avian schistosome within an urban environment, a location regularly visited by various wild waterfowl and the possible species of avian schistosomes responsible for cercarial dermatitis in Hungary. Taken as a whole, we demonstrate the actual and potential risk zone for cercarial dermatitis, particularly with reference to R. auricularia distributions, within Hungary and across Central Europe.

From Parasitized to Healthy-Looking Ants (Hymenoptera: Formicidae): Morphological Reconstruction Using Algorithmic Processing.

BACKGROUND: Parasites cause predictable alternative phenotypes of host individuals. Investigating these parasitogenic phenotypes may be essential in cases where parasitism is common or taxa is described based on a parasitized individual. Ignoring them could lead to erroneous conclusions in biodiversity-focused research, taxonomy, evolution, and ecology. However, to date, integrating alternative phenotypes into a set of wild-type individuals in morphometric analysis poses extraordinary challenges to experts. This paper presents an approach for reconstructing the putative healthy morphology of parasitized ants using algorithmic processing. Our concept enables the integration of alternative parasitogenic phenotypes in morphometric analyses. METHODS: We tested the applicability of our strategy in a large pool of Cestoda-infected and healthy individuals of three Temnothorax ant species (T. nylanderi, T. sordidulus, and T. unifasciatus). We assessed the stability and convergence of morphological changes caused by parasitism across species. We used an artificial neural network-based multiclass classifier model to predict species based on morphological trait values and the presence of parasite infection. RESULTS: Infection causes predictable morphological changes in each species, although these changes proved to be species-specific. Therefore, integrating alternative parasitogenic phenotypes in morphometric analyses can be achieved at the species level, and a prior species hypothesis is required. CONCLUSION: Despite the above limitation, the concept is appropriate. Beyond parasitogenic phenotypes, our approach can also integrate morphometric data of an array of alternative phenotypes (subcastes in social insects, alternative morphs in polyphenic species, and alternative sexes in sexually dimorphic species) whose integrability had not been resolved before.

Morphology of Pholeoixodes species associated with carnivores in the western Palearctic: Pictorial key based on molecularly identified Ixodes (Ph.) canisuga, I. (Ph.) hexagonus and I. (Ph.) kaiseri males, nymphs and larvae.

Three Palearctic members of the subgenus Pholeoixodes, i.e., Ixodes canisuga, Ixodes hexagonus and Ixodes kaiseri are frequently collected from dogs, cats, red foxes, badgers and other carnivorous/insectivorous hosts in Europe. While a pictorial identification key has been reported for female Pholeoixodes ticks, a similar work has not been done on their male, nymphal and larval specimens. This study was initiated in order to clarify and re-examine those morphological characters of these three tick species, which can be used relatively easily to identify/distinguish them. In the case of larvae the aims included finding alternatives to chaetotaxy, which is difficult to observe and its usefulness is also affected by uncertainties in literature data. For this, 609 Pholeoixodes ticks (males, nymphs and larvae) were collected from carnivores, hedgehogs and their environment in six European countries (representing Western, Central and Southeastern Europe), followed by detailed morphological examination and/or molecular analyses to confirm the identity of their species. Based on the morphology of 84 molecularly analyzed specimens and a new identification key compiled accordingly, altogether 116 I. canisuga, 277 I. hexagonus and 216 I. kaiseri males, nymphs and larvae were identified. Ixodes kaiseri was not found in Western Europe, where I. canisuga predominated. In Central Europe, all three Pholeoixodes species were collected, the largest number of specimens represented by I. hexagonus. On the other hand, in Southeastern Europe I. kaiseri had the highest abundance. In conclusion, the morphology of internal spur on the first coxae (as the traditionally used character to distinguish I. hexagonus from other Pholeoixodes species) is trustworthy to recognize males but is less informative in the case of nymphs and larvae. The latter can be identified more properly by observing the morphology of basis capituli. In particular, nymphs and larvae of I. canisuga have anteriorly flattened basis capituli, forming a plateau that surrounds the base of the hypostome. On the other hand, nymphs and larvae of I. hexagonus and I. kaiseri lack a similar plateau, but (unlike I. canisuga) have cornuae, which are either posterolaterally or caudally directed, respectively.

Malaria infection status of European Robins seems to associate with timing of autumn migration but not with actual condition.

Avian malaria parasites can negatively affect many aspects of the life of the passerines. Though these parasites may strongly affect the health and thus migration patterns of the birds also during autumn, previous studies on avian malaria focused mainly on the spring migration and the breeding periods of the birds. We investigated whether the prevalence of blood parasites varies in relation to biometrical traits, body condition and arrival time in the European Robin (Erithacus rubecula) during autumn migration. We found no sex or age related differences in avian malaria prevalence and no relationship between infection status and body size or actual condition of the birds was found either. However, the timing of autumn migration differed marginally between infected and non-infected juveniles, so that parasitized individuals arrived later at the Hungarian stopover site. This is either because avian malaria infections adversely affect the migration timing or migration speed of the birds, or because later arriving individuals come from more distant populations with possibly higher blood parasite prevalence. The possible delay that parasites cause in the arrival time of the birds during autumn migration could affect the whole migratory strategy and the breeding success of the birds in the next season.

Road-killed mammals provide insight into tick-borne bacterial pathogen communities within urban habitats.

Small- and medium-sized mammals play an important role in the life cycle of tick-borne pathogens in urban habitats. Our aim was to apply the general protocol, DAMA (documentation-assessment-monitoring-action), which is an integrated proposal to build a proactive capacity to understand, anticipate, and respond to the outcomes of accelerating environmental change. Here we tested whether road-killed carcasses in urban areas are useful sources of tissue and parasite samples to investigate these species' contribution to the epidemiology of vector-borne diseases. We collected 29 road-killed and 6 carcasses with different causes of mortality (23 northern white-breasted hedgehogs and 12 from seven other mammal species) mainly from Budapest, Hungary. We used quantitative and conventional PCRs to determine pathogens in 90 collected tissues (52 from hedgehogs; 38 from other species) and 417 ticks that were only found on hedgehogs. Tissue samples revealed a wide range of bacteria including human zoonotic pathogens identified as Anaplasma phagocytophilum ecotype I, Borrelia afzelii, B. spielmanii, Borrelia miyamotoi, Rickettsia helvetica, and Bartonella species. Among the 23 collected hedgehog carcasses, 17 (74%) were infected with A. phagocytophilum, 6 (26%) with Borrelia burgdorferi s.l., 12 (52%) with R. helvetica, and 15 (65%) with Rickettsia sp. Furthermore, we report the first detection of Rickettsia sp. infection in European moles and lesser weasel and R. helvetica in stone marten. Through sequencing B. afzelii, R. helvetica, R. monacensis and A. phagocytophilum ecotype I were identified in the ticks removed from the carcasses. We showed that road-killed urban mammal species are exposed to multiple tick-borne pathogens but further studies have to clarify whether they, in fact, also have a role in their maintenance and spread. Our study also demonstrates that roadkill can be used in the risk assessment of potential human infection and in the implementation of the DAMA protocol.

Investigations on the distribution of Schistosoma turkestanicum Skrjabin, 1913 (Trematoda: Schistosomatidae) infection of red deer in Hungary and a combined method for the detection of S. turkestanicum eggs in droppings.

Additional geographical distribution of the Central European populations of Schistosoma turkestanicum and the detectability of their eggs in droppings were investigated in red deer samples, because this rare species had previously been shown only in a single Hungarian habitat. Samples from visceral organs, intestinal contents, and droppings on the ground from 11 hunting areas of Hungary were investigated to find a new presence of this fluke. Close to the first site of detection in the Gemenc forest another habitat along the southern border of the country was found where the parasite lives in red deer. Therefore, it is possible that the worm also occurs in neighbouring Serbia or Croatia. Schistosoma turkestanicum causes a low-intensity infection in red deer and this host sheds low amounts of eggs, therefore the eggs are difficult to detect. Droppings were cleared by sedimentation, filtered by sieve screening and then the eggs were flotated using solutions with an increasing density of 1200 g/L, 1300 g/L, 1350 g/L, and 1400 g/L while they were being stained red with acid fuchsin. Eggs in fresh faeces can be most efficiently separated from plant fibres using a flotation solution of 1350 g/L density, but in some cases eggs in old dung can be detected using a solution of a specific gravity lower or higher than that. By combining the advantages of the three concentration processes, eggs of S. turkestanicum, which are more recognisable by the red stain, can be found in samples in which they are present at a density lower than 1/g.

High mitochondrial sequence divergence in synanthropic flea species (Insecta: Siphonaptera) from Europe and the Mediterranean.

BACKGROUND: Adult fleas are haematophagous ectoparasites of warm-blooded vertebrates, particularly mammals. Among them, the cat flea (Ctenocephalides felis) and the human flea (Pulex irritans) have high veterinary-medical significance, owing to their cosmopolitan distribution and role in the transmission of important vector-borne pathogens. While the taxonomy of Ct. felis has been investigated on a morphological basis during the past decades, its molecular-phylogenetic analyses have been only recently conducted. This study expands the knowledge on Ct. felis from hitherto less studied geographical regions, and includes representatives from additional flea families, less investigated with molecular approaches. METHODS: Fleas were collected in four countries of the Mediterranean Basin (Croatia, Italy, Malta and Israel), as well as in Hungary, from domestic and wild carnivores, rodents and humans. The DNA extracts of representative fleas (n = 148), belonging to ten species of eight genera, were used for PCR amplification of part of their cytochrome c oxidase subunits 1, 2 (cox1, cox2) and 18S rRNA genes, followed by sequencing and phylogenetic analyses. RESULTS: The majority (65.6%) of Ct. felis felis cox2 sequences showed 99.4-100% similarity to each other (haplogroup A), whereas those from Malta and Israel had 98.1-98.7% sequence similarity (haplogroup B), and a third sequence from Israel (haplotype C) had as low as 96.3% sequence similarity in comparison with a reference sequence from group "A". Except for the shape of the head, no consistent morphological differences (e.g. in chaetotaxy) were found between haplogroups "A" and "C". Haplotypes of Ct. canis were genetically more homogenous, with 99.6-100% sequence similarity to each other. However, when P. irritans collected from humans was compared to those from three species of wild carnivores, these only had 96.6% cox2 similarity. The mouse flea, Leptopsylla segnis and the northern rat flea, Nosopsyllus fasciatus were both shown to have haplotypes with low intraspecific cox2 similarities (96.2 and 94.4%, respectively). Taken together, differences between mitochondrial lineages within four flea species exceeded that observed between two Chaetopsylla spp. (which had 97.3% cox2 similarity). The topologies of cox1 and cox2 phylogenetic trees were in line with relevant sequence comparisons. Conversely, 18S rRNA gene analyses only resolved differences above the species level. CONCLUSIONS: Ctenocephalides felis felis, P. irritans, L. segnis and N. fasciatus were shown to have such a high level of mitochondrial gene heterogeneity, that the uniformity of these flea taxa should be reconsidered. Although the present results are limited (especially in the case of L. segnis and N. fasciatus), there appears to be no geographical or host restriction, which could explain the divergence of these genetic lineages.

An investigation of the host-specificity of metacercariae of species of Apophallus (Digenea: Heterophyidae) in freshwater fishes using morphological, experimental and molecular methods.

Metacercariae of species of the genus Apophallus Lühe, 1909, infecting the fins and skin of freshwater fishes, frequently cause black spot disease. Two species, Apophallus muehlingi (Jägerskiöld, 1899) and A. donicus (Skrjabin & Lindtrop, 1919), are known to occur in Hungarian fishes. It has generally been thought that metacercariae of A. muehlingi infect cyprinid fishes, whereas those of A. donicus develop in percids. As part of a morphological, experimental and molecular study, metacercariae were collected from 99 infected specimens of five cyprinid hosts (Abramis brama, Blicca bjoerkna, Chondrostoma nasus, Squalius cephalus, Scardinius erythrophthalmus) and 18 infected specimens of two percid hosts (Gymnocephalus cernua, Perca fluviatilis) in Hungarian natural waters (Lake Balaton, River Danube). Moreover, 1024 common carp (Cyprinus carpio) specimens collected from Hungarian fish ponds were investigated for Apophallus infection, but without positive results. For reliable species identification, experimental infections of chicks were carried in order to produce adult specimens from metacercariae collected from the fins and skin of the cyprinid and percid hosts. Within 8 days, adult specimens of both A. muehlingi and A. donicus developed in chicks infected with metacercariae from the cyprinid common bream (Abramis brama) and the white bream (Blicca bjoerkna) and the ruffe (Gymnocephalus cernua), a percid, respectively. The morphology of the collected metacercariae and adult individuals developed in the feeding experiments was characterised. A molecular analysis was extended to cercarial samples from the snail Lithoglyphus naticoides and to a single adult specimen of Apophallus from a fox. Sequences of 28 specimens were analysed using molecular methods (sequencing the internal transcribed spacer region and the cytochrome oxidase I subunit). Phylogenetic analysis was executed, and the Apophallus samples clustered into three distinct branches using both genes, A. muehlingi from cyprinids, A. donicus from percids and, a third, previously unknown, Apophallus clade, also from cyprinids.

Signatures of mito-nuclear discordance in Schistosoma turkestanicum indicate a complex evolutionary history of emergence in Europe.

High levels of molecular diversity were identified in mitochondrial cytochrome c oxidase (cox1) gene sequences of Schistosoma turkestanicum from Hungary. These cox1 sequences were all specific to Hungary which contrasted with the low levels of diversity seen in the nuclear internal transcribed spacer region (ITS) sequences, the majority of which were shared between China and Iran isolates. Measures of within and between host molecular variation within S. turkestanicum showed there to be substantial differences in molecular diversity, with cox1 being significantly more diverse than the ITS. Measures of haplotype frequencies revealed that each host contained its own subpopulation of genetically unique parasites with significant levels of differentiation. Pairwise mismatch analysis of cox1 sequences indicated S. turkestanicum populations to have a bimodal pairwise difference distribution and to be stable unlike the ITS sequences, which appeared to have undergone a recent population expansion event. Positive selection was also detected in the cox1 sequences, and biochemical modelling of the resulting protein illustrated significant mutational events causing an alteration to the isoelectric point of the cox1 protein, potentially altering metabolism. The evolutionary signature from the cox1 indicates local adaptation and long establishment of S. turkestanicum in Hungary with continual introgression of nuclear genes from Asian isolates. These processes have led to the occurrence of mito-nuclear discordance in a schistosome population.

Lethal Angiostrongylus vasorum infection in a Hungarian dog.

During post-mortem examination of lungs and heart of a 7-month-old female French bulldog, 158 worms were collected from the lung vessels and they proved Angiostrongylus vasorum by their morphological and genetic identification by PCR. The histopathological investigation found a multifocal interstitial inflammation characterized by numerous lymphocytes and a smaller number of plasma cells and eosinophils whilst L1 stage larvae could be seen inside dilated alveoli. We suggested a lethal angiostrongylosis supposed to lead to a fatal effect. Our report attracts attention to the presence of the nematode A. vasorum as causative agent of canine cardiopulmonary disorder in the south-western region of Hungary.

[A rare zoonosis in Hungary: cercarial dermatitis caused by Schistosoma turkestanicum blood-fluke]. / Egy ritka zoonosis: a Schistosoma turkestanicum vérmétely által okozott cercaria dermatitis Magyarországon.

Several trematodes that parasitize vertebrate animals utilize swimming aquatic larvae to infect the host percutaneously. The most important ones among these parasites are the blood-flukes of birds and mammals comprising species that are also zoonotic. Within this latter group are species that cause the bilharziasis or schistosomiasis of inhabitants of the tropical countries, and other trematode species that are able to penetrate human skin, but do not develop to an adult form of the worm in the body. In temperate climates this latter type of infection occurs mainly in the form of an unpleasant inflammation of the skin and is often called "swimmer's itch". In most of these cases, the origin of the larvae remains unexplored, the source of the infection is neglected by the medical or veterinarian practitioners. Herein we report for the first time in Hungary that the cause of such dermatitis was the cercariae of Schistosoma turkestanicum, which infected red deer (Cervus elaphus) in this country. The local name of this pristine disease is "water mange" and it occurs only in one of the floodplains of the Danube. On the basis of informal communication this symptom seems to be rather regular among people who do fishing or have a bath in the habitat of the blood-fluke. In the case of adequate anamnesis it is worth examining the origin of the cercarial dermatitis which may give cross-reactions with human schistosomiasis during serological tests. Orv. Hetil., 2016, 157(40), 1579-1586.

Dermacentor reticulatus: a vector on the rise.

Dermacentor reticulatus is a hard tick species with extraordinary biological features. It has a high reproduction rate, a rapid developmental cycle, and is also able to overcome years of unfavourable conditions. Dermacentor reticulatus can survive under water for several months and is cold-hardy even compared to other tick species. It has a wide host range: over 60 different wild and domesticated hosts are known for the three active developmental stages. Its high adaptiveness gives an edge to this tick species as shown by new data on the emergence and establishment of D. reticulatus populations throughout Europe. The tick has been the research focus of a growing number of scientists, physicians and veterinarians. Within the Web of Science database, more than a fifth of the over 700 items published on this species between 1897 and 2015 appeared in the last three years (2013-2015). Here we attempt to synthesize current knowledge on the systematics, ecology, geographical distribution and recent spread of the species and to highlight the great spectrum of possible veterinary and public health threats it poses. Canine babesiosis caused by Babesia canis is a severe leading canine vector-borne disease in many endemic areas. Although less frequently than Ixodes ricinus, D. reticulatus adults bite humans and transmit several Rickettsia spp., Omsk haemorrhagic fever virus or Tick-borne encephalitis virus. We have not solely collected and reviewed the latest and fundamental scientific papers available in primary databases but also widened our scope to books, theses, conference papers and specialists colleagues' experience where needed. Besides the dominant literature available in English, we also tried to access scientific literature in German, Russian and eastern European languages as well. We hope to inspire future research projects that are necessary to understand the basic life-cycle and ecology of this vector in order to understand and prevent disease threats. We conclude that although great strides have been made in our knowledge of the eco-epidemiology of this species, several gaps still need to be filled with basic research, targeting possible reservoir and vector roles and the key factors resulting in the observed geographical spread of D. reticulatus.

Identification of Hepatozoon erhardovae Krampitz, 1964 from bank voles (Myodes glareolus) and fleas in Southern Hungary.

In order to investigate the prevalence and life cycle of apicomplexan parasites, small mammals were live-trapped with modified Sherman traps in Southern Hungary between 2010 and 2012. Altogether, 528 rodents (Apodemus flavicollis Melchior, 1834, Apodemus agrarius Pallas, 1771, Myodes glareolus Schreber, 1780, Microtus agrestis Linnaeus, 1761, Mus musculus Linnaeus, 1758 and Micromys minutus Pallas, 1771) were collected and four shrews (Sorex spp.) were by-catched. Captured animals belonging to non-protected species were euthanized, and spleen samples were preserved for histological and molecular analyses. During the examination of spleen smears, Hepatozoon parasites were observed in eight out of 48 bank voles (M. glareolus). DNA was isolated from altogether 221 spleen samples, and 18S rDNA was amplified using two different PCR protocols. The eight bank vole samples were positive with PCR, but none of the other M. glareolus spleen samples or any of the tissue samples from other species were found to be infected. Sequenced amplicons were very similar to Hepatozoon spp. detected in M. glareolus in Spain and Poland. Ectoparasites were collected from the small mammal carcasses and from the vegetation. Hepatozoon DNA was not found in the 181 ticks removed from the small mammals or in the 162 ticks collected with flagging, but was detected in all three flea species (4/43 Megabothris turbidus Rothschild, 1909, 3/10 Ctenophthalmus assimilis Taschenberg, 1880 and 7/78 Ctenophthalmus agyrtes Heller, 1896). Based on gamont morphology, vertebrate and arthropod host species and DNA sequences, the parasites in our study can be identified as Hepatozoon erhardovae.

Prevalence and diversity of human pathogenic rickettsiae in urban versus rural habitats, Hungary.

Tick-borne rickettsioses belong to the important emerging infectious diseases worldwide. We investigated the potential human exposure to rickettsiae by determining their presence in questing ticks collected in an urban park of Budapest and a popular hunting and recreational forest area in southern Hungary. Differences were found in the infectious risk between the two habitats. Rickettsia monacensis and Rickettsia helvetica were identified with sequencing in questing Ixodes ricinus, the only ticks species collected in the city park. Female I. ricinus had a particularly high prevalence of R. helvetica (45%). Tick community was more diverse in the rural habitat with Dermacentor reticulatus ticks having especially high percentage (58%) of Rickettsia raoultii infection. We conclude that despite the distinct eco-epidemiological traits, the risk (hazard and exposure) of acquiring human pathogenic rickettsial infections in both the urban and the rural study sites exists.

Eco-epidemiology of Borrelia miyamotoi and Lyme borreliosis spirochetes in a popular hunting and recreational forest area in Hungary.

BACKGROUND: Borrelia miyamotoi, the newly discovered human pathogenic relapsing fever spirochete, and Borrelia burgdorferi sensu lato are maintained in natural rodent populations. The aim of this study was to investigate the natural cycle of B. miyamotoi and B. burgdorferi s.l. in a forest habitat with intensive hunting, forestry work and recreational activity in Southern Hungary. METHODS: We collected rodents with modified Sherman-traps during 2010-2013 and questing ticks with flagging in 2012. Small mammals were euthanized, tissue samples were collected and all ectoparasites were removed and stored. Samples were screened for pathogens with multiplex quantitative real-time polymerase chain reaction (qPCR) targeting a part of flagellin gene, then analysed with conventional PCRs and sequencing. RESULTS: 177 spleen and 348 skin samples of six rodent species were individually analysed. Prevalence in rodent tissue samples was 0.2 % (skin) and 0.5 % (spleen) for B. miyamotoi and 6.6 % (skin) and 2.2 % (spleen) for B. burgdorferi s.l. Relapsing fever spirochetes were detected in Apodemus flavicollis males, B. burgdorferi s.l. in Apodemus spp. and Myodes glareolus samples. Borrelia miyamotoi was detected in one questing Ixodes ricinus nymph and B. burgdorferi s.l in nymphs and adults. In the ticks removed from rodents DNA amplification of both pathogens was successful from I. ricinus larvae (B. miyamotoi 5.6 %, B. burgdorferi s.l. 11.1 %) and one out of five nymphs while from Ixodes acuminatus larvae, and nymph only B. burgdorferi s.l. DNA was amplified. Sequencing revealed B. lusitaniae in a questing I. ricinus nymph and altogether 17 B. afzelii were identified in other samples. Two Dermacentor marginatus engorged larva pools originating from uninfected hosts were also infected with B. afzelii. CONCLUSIONS: This is the first report of B. miyamotoi occurrence in a natural population of A. flavicollis as well as in Hungary. We provide new data about circulation of B. burgdorferi s.l. in rodent and tick communities including the role of I. acuminatus ticks in the endophilic pathogen cycle. Our results highlight the possible risk of infection with relapsing fever and Lyme borreliosis spirochetes in forest habitats especially in the high-risk groups of hunters, forestry workers and hikers.

Angiostrongylosis-related restrictive pneumopathy assessed by arterial blood gas analysis in a dog.

Pulmonary angiostrongylosis was diagnosed by the Baermann method and larval identification from faecal and bronchoalveolar lavage samples in a five-month- old male mongrel dog with dyspnoea and cough. Arterial blood gas analysis indicated arterial hypoxaemia and restrictive pneumopathy. In addition to the palliative treatment, fenbendazole was administered (50 mg/kg/24 h per os) for 14 days. The respiratory signs subsided within a short time clinically, but serial arterial blood gas analysis demonstrated an ongoing ventilation disorder. Repeated haematology, thoracic radiography, bronchoscopy and blood gas analysis were performed to follow the course of the disease. The most severe eosinophilia was detected after the beginning of the anthelmintic therapy, and the arterial pO2 level was permanently low. Arterial blood gas analysis provided the most adequate information about the course of the pneumopathy and it greatly facilitated the patient's medical management.

Sarcocystis-infection of cattle in Hungary.

BACKGROUND: Reports on Sarcocystis-infection of cattle are outdated or lacking in many European countries, including those in the Central-Eastern part of the continent. Therefore, to assess the prevalence of Sarcocystis spp. among bovids in Hungary, a countrywide survey was initiated. In addition, fulminant deaths of four cattle, that showed clinical signs and post mortem lesions resembling acute sarcocystiosis ("Dalmeny disease"), were investigated. METHODS: During the countrywide survey individual heart and oesophagus samples were collected at slaughterhouses from 151 beef cattle and from 15 buffalo, kept in 31 places of Hungary. Analysis for Sarcocystis spp. was carried out with conventional PCRs for the 18S rDNA gene and gel electrophoresis, followed by sequencing of 36 strongly positive samples. Mortality cases were evaluated by histological, molecular, bacteriological and virological analyses of samples from various organs. RESULTS: Among slaughtered cattle the rate of Sarcocystis-infection was 66%. S. cruzi was identified as the most prevalent species in aurochs-like breed, and the zoonotic S. hominis in Hungarian grey cattle. Concerning the sudden deaths of cattle, Sarcocystis-infection could not be demonstrated in organs showing haemorrhages, but S. cruzi cysts were present in the muscles. In one case "S. sinensis" was molecularly identified in the blood (indicating sarcocystaemia). Results of analyses for bacterial/viral pathogens were negative. CONCLUSIONS: S. cruzi appears to be the most prevalent Sarcocystis sp. in cattle in Hungary, followed by the zoonotic S. hominis. However, the rate of infection with both species was shown to differ between cattle breeds. The suspected role of Sarcocystis spp. as causative agents of the fatal cases could not be confirmed.

Candidatus Neoehrlichia mikurensis and Anaplasma phagocytophilum in natural rodent and tick communities in Southern Hungary.

The aim of this study was to investigate the natural cycle of the new human pathogenic bacteria Candidatus Neoehrlichia mikurensis and Anaplasma phagocytophilum in Southern Hungary. We collected rodents with live-traps (2010-2013) and questing ticks with flagging in 2012. Small mammals were euthanized, tissue samples were collected and all the ectoparasites were removed and stored in 70% alcohol. We found relatively low overall prevalence of tick infestation (8%). Samples were analysed for A. phagocytophilum and Candidatus N. mikurensis with multiplex quantitative real-time PCR targeting a part of major surface protein 2 (msp2) and the heat shock protein groEL genes, respectively. The overall prevalence in tissue samples was 6.6% (skin) and 5.1% (spleen) for A. phagocytophilum and 1.7% (skin) and 3.4% (spleen) for Candidatus N. mikurensis. Candidatus N. mikurensis was only detected in Apodemus flavicollis and Apodemus agrarius, while A. phagocytophilum was found in A. flavicollis, A. agrarius, Myodes glareolus, Microtus arvalis and Mus musculus samples. Prevalence of A. phagocytophilum in skin samples of A. flavicollis was significantly higher than prevalence of N. mikurensis (p<0.05). Among questing Ixodes ricinus ticks we found three (8.8%) individuals (female, male, nymph) infected with Candidatus N. mikurensis. Five (3.1%) questing ticks had A. phagocytophilum infection (one I. ricinus male, two Dermacentor reticulatus females and two Haemaphysalis concinna females). We found one I. ricinus nymph removed from a male A. flavicollis with A. phagocytophilum infection. Our study provides new data on the occurrence of these pathogens in rodent tissue samples, questing ticks and engorged ticks in Southern Hungary.