Case report: First report of potentially zoonotic Gongylonema pulchrum in a free-living roe deer (Capreolus capreolus) in Slovenia.

Adult female and male Gongylonema nematodes were found in the oesophagus of a free-living roe deer (Capreolus capreolus) in Slovenia during passive health surveillance of wildlife. The genus Gongylonema was determined by light microscopy based on the genus-specific cuticular bosses in the anterior part of the parasite. Molecular methods were used to confirm the species Gongylonema pulchrum, which has zoonotic potential. Although Gongylonema species are considered common and distributed worldwide, this is the first report of G. pulchrum in an animal on the territory of Slovenia and the first molecular report in a roe deer worldwide. The parasite is likely to be underdiagnosed, misdiagnosed or goes unnoticed as the animals show little or no clinical signs and minor pathological lesions. Slaughterhouse workers, hunters and veterinarians should be aware of this elusive parasite. Examination and evisceration of the upper digestive tract of animals should therefore be carried out more carefully.

Prion protein gene (PRNP) variation in German and Danish cervids.

The structure of cellular prion proteins encoded by the prion protein gene (PRNP) impacts susceptibility to transmissible spongiform encephalopathies, including chronic wasting disease (CWD) in deer. The recent emergence of CWD in Northern European reindeer (Rangifer tarandus), moose (Alces alces alces) and red deer (Cervus elaphus), in parallel with the outbreak in North America, gives reason to investigate PRNP variation in European deer, to implement risk assessments and adjust CWD management for deer populations under threat. We here report PRNP-sequence data from 911 samples of German red, roe (Capreolus capreolus), sika (Cervus nippon) and fallow deer (Dama dama) as well as additional data from 26 Danish red deer close to the German border and four zoo species not native to Germany. No PRNP sequence variation was observed in roe and fallow deer, as previously described for populations across Europe. In contrast, a broad PRNP variation was detected in red deer, with non-synonymous polymorphisms at codons 98, 226 and 247 as well as synonymous mutations at codons 21, 78, 136 and 185. Moreover, a novel 24 bp deletion within the octapeptide repeat was detected. In summary, 14 genotypes were seen in red deer with significant differences in their geographical distribution and frequencies, including geographical clustering of certain genotypes, suggesting "PRNP-linages" in this species. Based on data from North American CWD and the genotyping results of the European CWD cases, we would predict that large proportions of wild cervids in Europe might be susceptible to CWD once introduced to naive populations.

Nasopharyngeal Botflies Oestrus ovis and Cephenemyia stimulator in Roe Deer (Capreolus capreolus) from Northern Spain.

Two of 19 roe deer (Capreolus capreolus) in northern Spain were infested with Cephenemyia stimulator and one individual with Oestrus ovis. Three larvae of O. ovis were recovered from the nasopharynx of an adult female deer examined in February 2017, during the hunting season in the province of Burgos. The larvae were identified by morphology, and DNA sequencing from cytochrome oxidase subunit I and 28S rDNA confirmed the identification of both species of larvae. The possibility of O. ovis parasitizing roe deer should be considered in the diagnosis of nasal bots in cervids.

Shiga Toxin-Producing Escherichia coli Isolated from Wild Ruminants in Liguria, North-West Italy.

Wildlife may represent an important source of infectious diseases for humans and other wild and domestic animals. Wild ruminants can harbour and transmit Shiga toxin-producing Escherichia coli (STEC) to humans, and some strains even carry important antimicrobial resistance. In this study, 289 livers of wild roe deer, fallow deer, red deer and chamois collected in Liguria, north-west Italy, from 2019 to 2023 were analysed. Overall, 44 STEC strains were isolated from 28 samples. The characterisation of serogroups showed the presence of O104, O113, O145 and O146 serogroups, although for 28 colonies, the serogroup could not be determined. The most prevalent Shiga toxin gene in isolated strains was Stx2, and more specifically the subtype Stx2b. The other retrieved subtypes were Stx1a, Stx1c, Stx1d and Stx2g. The isolated strains generally proved to be susceptible to the tested antimicrobials. However, multi-drug resistances against highly critical antimicrobials were found in one strain isolated from a roe deer. This study highlights the importance of wildlife monitoring in the context of a "One Health" approach.

Seroprevalence of infectious pathogens of zoonotic and veterinary importance in wild ruminants from Slovenia.

Introduction: Wildlife represents an increasingly important source of pathogens of medical and veterinary importance. Surveillance in wildlife offers an insight on current epidemiological status of selected pathogens and help to prevent spillovers to humans and livestock. Material and methods: Our study included 312 wild ruminants belonging to five species: Roe deer (n = 134), red deer (n = 113), Alpine chamois (n = 53), European mouflon (n = 10) and Alpine ibex (n = 2). Seven pathogens that may have profound effect on human/livestock health and economic viability of the farms were tested using serological methods. Results: Antibodies against Toxoplasma gondii, Neospora caninum, Coxiella burnetii, Brucella spp., Chlamydophila abortus, Mycobacterium avium subsp. paratuberculosis (MAP) and Mycobacterium bovis were detected in 34.62% (108/312), 0.96% (3/312), 2.24% (7/312), 0, 0.96% (3/312), 0, 0.64% (2/312) of animals tested, respectively. Because of low prevalences, risk factors were assessed only for T. gondii. Sex (female>male) and species (roe deer>red deer, roe deer>Alpine chamois) were significantly associated with the T. gondii positive outcome, while age was not. Discussion: Adult males had the lowest T. gondii prevalence which offers future research opportunities. The lower seroprevalence of most investigated pathogens suggests game meat, if properly cooked, as being relatively safe for human consumption. This is the first study investigating the seroprevalence and associated risk factors of selected pathogens in wild ruminants in Slovenia.

Dissecting the Effects of Cephenemyia stimulator on the Olfactory Turbinates and Nasopharynx of Roe Deers (Capreolus capreolus).

Nasopharyngeal myiasis in European roe deer (Capreolus capreolus) is a pathological condition caused by the larval stages of Cephenemyia stimulator, a fly from the Oestridae family. These larvae reside in the host's upper respiratory tract for months, inducing significant tissue damage and clinical symptoms. The lifecycle of Cephenemyia stimulator is complex, involving three larval stages before maturation into adult flies, with each stage contributing to the progressive pathology observed in the host. Despite their prevalence, the histopathological effects of these larvae in the nasal and nasopharyngeal cavities have been understudied. Our study fills this knowledge gap by providing a detailed histopathological analysis of the affected tissues, using various staining techniques to reveal the extent and nature of the damage caused by these parasitic larvae. This histopathological examination reveals significant alterations within the nasopharyngeal mucosa and nasal cavity, including erythematous changes, mucosal metaplasia, fibrosis, and tissue necrosis. Parasitic cysts and eosinophilic infiltration further characterize the impact of the infestation, compromising not only the mucosal integrity but also potentially the olfactory function of the affected animals. This research is crucial for understanding the impact of myiasis on both the health and olfactory capabilities of roe deer populations and could have significant implications for wildlife management and conservation.

First Report of Caseous Lymphadenitis by Corynebacterium pseudotubercolosis and Pulmonary Verminosis in a Roe Deer (Capreolus capreolus Linnaeus, 1758) in Italy.

Caseous lymphadenitis is a chronic debilitating disease typical of small ruminants, but it is also noted in several other domestic and wild species. In this report, we present the first documented case in Italy of pseudotuberculosis in a roe deer (Capreolus capreolus, Linnaeus 1758) found dead in the mountains of Forlì-Cesena province, Emilia Romagna region. The carcass underwent necropsy according to standard protocols, revealing generalized lymphadenopathy and severe apostematous pneumonia with multifocal and encapsulated abscesses. Corynebacterium pseudotuberculosis was isolated from the lung parenchyma, lymph nodes and abscesses. Additionally, severe parasitic bronchopneumonia of the caudal lobes and gastrointestinal strongyle infestation were detected. To our knowledge, this is the first documented case of CLA referable to C. pseudotubercolosis in a roe deer in Italy.

Cadmium in Selected Organs of Game Animals from Areas with Different Degrees of Industrialisation and Its Intake by Human Consumers.

The aim of this study was to determine the concentration of cadmium (Cd) in the organs of game animals from areas with different degrees of industrialisation and to assess its intake following the consumption of the livers of these animals. The material for this study consisted of liver and kidney samples taken from roe deer (Capreolus capreolus), red deer (Cervus elaphus), and wild boar (Sus scrofa) from five regions differing in land use and industrialisation. Compared to the kidneys, the liver was an indicator of the current state of the environment and Cd intake, while the kidney was used to assess the long-term accumulation of Cd in the body. The cadmium concentration in the samples was determined by inductively coupled plasma atomic emission spectrometry. The cadmium concentrations of the organs were found to vary according to geographical location, with the highest levels being observed in Southern Poland, i.e., areas characterised by the highest degree of industrialisation and the presence of heavy industry. However, high Cd concentrations were also reported in the northeastern region. Examples of excessive cadmium levels in the kidneys were noted in all tested regions. Cd intake by humans was evaluated by calculating the estimated daily cadmium intake, and this was compared with the tolerable weekly intake (TWI). The TWI was only exceeded by children frequently consuming wild boar liver from the southwest region.

Wildlife-vehicle collisions: The disproportionate risk of injury faced by motorcyclists.

We applied a generalized linear mixed-effects model to determine the factors leading to injuries from wildlife-vehicle collisions (WVCs). We used the Police database representing WVCs which took place on the Czech road network between 2009 and 2022. The majority of WVCs in Czechia are with roe deer, followed by wild boar, i.e., both relatively small ungulates. Less than 2 % of these encounters ends with an injury to the motor vehicle occupants. We found that the probability of sustaining injury was systematically higher for motorcyclists than for car occupants. The odds of sustaining an injury during WVC were roughly 1600 times higher for motorcyclists than for car occupants. When applying an evading manoeuvre, the odds of sustaining an injury were approximately 68 times higher for car occupants while only 2.3 times higher for motorcyclists compared to a direct hit to an animal. The lack of helmets (for motorcyclists) and missing seat belts (for car occupants) were additional factors which made the outcomes worse for WVCs. While the acceptance of a direct hit (preceded by braking) seems to be a reasonable strategy for car drivers, WVC awareness (including maintaining a lower speed during critical times and places) should be raised among motorcyclists as both manoeuvres are almost comparably dangerous for them.

First Molecular Identification of Calicophoron daubneyi (Dinnik, 1962) and Paramphistomum leydeni (Nasmark, 1937) in Wild Ruminants from Romania.

Rumen flukes are geographically widespread trematodes affecting wild and domestic ruminants. The juvenile forms, which are found in the small intestine, are more pathogenic compared to the adults. Severe diarrhoea and weight loss are the major clinical signs, and the disease might be fatal in severely infested individuals. In the last decade, paramphistomosis has been described as an emerging parasitic disease in Europe. This study aimed to identify the rumen fluke species in wild ruminants from western Romania. Fifty-two pre-stomachs obtained from roe deer (Capreolus capreolus) that were hunted on 14 hunting grounds from Timiș and Arad counties were examined for the presence of paramphistomes. Three (9.09%) out of 33 samples were positive in Timiș County, and one (5.26%) out of 19 samples was positive in Arad County. Subsequent PCR testing revealed that three samples were positive for Calicophoron daubneyi and one for Paramphistomum leydeni. The presence of C. daubneyi and P. leydeni in roe deer has not been previously reported in Romania. Two Paramphistomum species-C. daubneyi and P. leydeni-were revealed as the main species of rumen flukes in roe deer from forests in Romania.

Contribution to the Knowledge of Gastrointestinal Nematodes in Roe Deer (Capreolus capreolus) from the Province of León, Spain: An Epidemiological and Molecular Study.

A study of gastrointestinal nematodes in roe deer was carried out in the regional hunting reserves of Riaño and Mampodre, Province of León, Spain, to provide information on their prevalence and intensity of infection in relation to the sampling areas, age of the animals, and body weight. Through a regulated necropsy of the animals, all of them harbored gastrointestinal nematodes in their digestive tract, with a mean intensity of parasitism of 638 ± 646.1 nematodes/infected animal. Eleven genera were found and 18 species of gastrointestinal nematodes were identified, three of them polymorphic: Trichostrongylus axei, Trichostrongylus vitrinus, Trichostrongylus capricola, Trichostrongylus colubriformis, Haemonchus contortus, Spiculopteragia spiculoptera/Spiculopteragia mathevossiani, Ostertagia leptospicularis/Ostertagia kolchida, Ostertagia (Grosspiculopteragia) occidentalis, Teladorsagia circumcincta/Teladorsagia trifurcate, Marshallagia marshalli, Nematodirus europaeus, Cooperia oncophora, Capillaria bovis, Oesophagostomum venulosum, and Trichuris ovis. All of them have already been cited in roe deer in Europe, but Marshallagia marshalli, Capillaria bovis, and Ostertagia (Grosspiculopteragia) occidentalis are reported for the first time in Spain in this host. The abomasum was the intestinal section, where the prevalence (98.9%) and mean intensity (x¯ = 370.7 ± 374.4 worms/roe deer; range 3-1762) were significantly higher, but no statistically significant differences were found when comparing the sampling areas and age of animals. The animals with lower body weight had a higher parasite load than those in better physical condition, finding, in this case, statistically significant differences (p = 0.0020). Seven genera and 14 species were identified. In the small intestine, 88% of the animals examined presented gastrointestinal nematodes, with an average intensity of x¯ = 131.7 ± 225.6 parasites/infected animal, ranging between 4-1254 worms. No statistically significant differences were found when the three parameters studied were compared. Four genera and seven species were identified. In the large intestine/cecum, 78.3% of the examined roe deer presented adult worms, with an average intensity of 6.3 ± 5.5 worms/infected animal; range 1-26 worms. Only statistically significant differences were observed when considering the mean intensity of parasitism and the sampling area (p = 0.0093). Two genera and two species were identified. Several of the species found in the study were studied molecularly, and with the sequences obtained compared with those deposited in GenBank, phylogenetic trees were prepared to determine their taxonomic status. Using coprological techniques, the existing correlation in the shedding of gastrointestinal nematode eggs in roe deer was investigated with that of semi-extensive sheep farms in the same study area to verify the existence of cross-transmission of these parasites between wild and domestic animals. The high values found in the studied parameters show that northern Spain is an area of high-intensity infection for roe deer.

Microbiota and Nutrient Portraits of European Roe Deer (Capreolus capreolus) Rumen Contents in Characteristic Southern German Habitats.

Roe deer (Capreolus capreolus) are found in various habitats, from pure forest cultures to agricultural areas and mountains. In adapting to the geographically and seasonally differentiating food supply, they depend, above all, on an adapted microbiome. However, knowledge about the microbiome of wild ruminants still needs to be improved. There are only a few publications for individual species with a low number of samples. This study aims to identify a core microbiota for Bavarian roe deer and present nutrient and microbiota portraits of the individual habitat types. This study investigated the roe deer's rumen (reticulorumen) content from seven different characteristic Bavarian habitat types. The focus was on the composition of nutrients, fermentation products, and the rumen bacterial community. A total of 311 roe deer samples were analysed, with the most even possible distribution per habitat, season, age class, and gender. Significant differences in nutrient concentrations and microbial composition were identified for the factors habitat, season, and age class. The highest crude protein content (plant protein and microbial) in the rumen was determined in the purely agricultural habitat (AG), the highest value of non-fibre carbohydrates in the alpine mountain forest, and the highest fibre content (neutral detergent fibre, NDF) in the pine forest habitat. Maximum values for fibre content go up to 70% NDF. The proportion of metabolites (ammonia, lactate, total volatile fatty acids) was highest in the Agriculture-Beech-Forest habitat (ABF). Correlations can be identified between adaptations in the microbiota and specific nutrient concentrations, as well as in strong fluctuations in ingested forage. In addition, a core bacterial community comprising five genera could be identified across all habitats, up to 44% of total relative abundance. As with all wild ruminants, many microbial genera remain largely unclassified at various taxonomic levels. This study provides a more in-depth insight into the diversity and complexity of the roe deer rumen microbiota. It highlights the key microorganisms responsible for converting naturally available nutrients of different botanical origins.

Helminth fauna in roe deer (Capreolus capreolus Linnaeus, 1758) in the province of Grosseto (central Italy).

Helminth infection was analysed at necropsy and coprology in a total of 54 roe deer from the province of Grosseto (central Italy) between 2018 - 2020. Age and sex data were recorded for each deer for a total of 31 adults (23 females, 8 males) and 23 juveniles (11 females, 12 males). The results on the small intestine (51 samples) highlighted that nematodes belonging to the species Trichostrongylus colubriformis were the most prevalent parasite (41.2 %), followed by the cestode Moniezia expansa (7.8 %). The large intestine results (52 samples) showed Trichuris spp. (53.8 %), Oesophagostomum venulosum (50 %) and Chabertia ovina (26.9 %). In the abomasum, only Ostertagia ostertagi (17.9 %) was found. Of the 34 samples analysed by bronchopulmonary, only the lung of an adult female was positive for Dictyocaulus spp. In two livers out of 33 samples analysed, nematodes of the species Setaria tundra were found on the surface. Copropositivity was observed in 45 of the 52 faecal samples analysed. The results of the present study indicate that the roe deer is host to several species of parasites, which are also common in other cervids and domestic ruminants. Statistical testing highlighted a significant difference between mean intensities in males and females.

Analysis of Some Biochemical Properties of Recombinant Siberian Roe Deer (Capreolus pygargus) Chymosin Obtained in the Mammalian Cell Culture (CHO-K1).

Structure of the chymosin gene of Siberian roe deer (Capreolus pygargus) was established for the first time and its exon/intron organization was determined. Coding part of the chymosin gene of C. pygargus was reconstructed by the Golden Gate method and obtained as a DNA clone. Comparative sequence analysis of the roe deer, cow, and one-humped camel prochymosins revealed a number of amino acid substitutions at the sites forming the substrate-binding cavity of the enzyme and affecting the S4 and S1' + S3' specificity subsites. Integration vector pIP1 was used to construct a plasmid pIP1-Cap in order to express recombinant roe deer prochymosin gene in CHO-K1 cells. CHO-K1-CYM-Cap pool cells were obtained, allowing synthesis and secretion of recombinant prochymosin into the culture fluid. As a result of zymogen activation, a recombinant roe deer chymosin was obtained and its total milk-clotting activity was estimated to be 468.4 ± 11.1 IMCU/ml. Yield of the recombinant roe deer chymosin was 500 mg/liter or ≈468,000 IMCU/liter, which exceeds the yields of genetically engineered chymosins in most of the expression systems used. Basic biochemical properties of the obtained enzyme were compared with the commercial preparations of recombinant chymosins from one-humped camel (Camelus dromedarius) and cow (Bos taurus). Specific milk-clotting activity of the recombinant chymosin of C. pygargus was 938 ± 22 IMCU/mg, which was comparable to that of the reference enzymes. Non-specific proteolytic activity of the recombinant roe deer chymosin was 1.4-4.5 times higher than that of the cow and camel enzymes. In terms of coagulation specificity, recombinant chymosin of C. pygargus occupied an intermediate position between the genetically engineered analogs of B. taurus and C. dromedarius chymosins. Thermostability threshold of the recombinant roe deer chymosin was 55°C. At 60°C, the enzyme retained <1% of its initial milk-clotting activity, and its complete thermal inactivation was observed at 65°C.

Winter activity of Ixodes ricinus in Sweden.

BACKGROUND: In Europe, Ixodes ricinus (Acari: Ixodidae) is the most widespread and abundant tick species, acting as a vector for several microorganisms of medical and veterinary importance. In Northern and Central Europe, the tick has a bimodal activity pattern consisting of a peak in spring to the beginning of summer and a second peak at the end of summer. However, several findings of ticks on animals during winter have been reported, which raises the question of whether this is an overwintering strategy or whether ticks are active during winter in Scandinavia. The objectives of our study were to determine (i) whether ticks were active and finding hosts during winter, (ii) whether they parasitize their hosts, and (iii) what climatic factors-i.e., temperature, snow depth and precipitation-govern tick winter activity. METHODS: Throughout three winter seasons, we examined wild-living and free-ranging roe deer (Capreolus capreolus) for ticks on 332 occasions. In total, 140 individual roe deer were captured in two climatically contrasting sites in south-central Sweden, Grimsö and the Bogesund research area, respectively. We re-examined individual roe deer up to 10 times within the same winter or approximately once a week (mean 10 days, median 7 days between re-examinations) and recorded the absence or presence of ticks on the animals, and tested to what extent meteorological factors affected tick activity. To determine the attachment day, we used the coxal/scutal index of 18 nymphs and 47 female ticks. RESULTS: In total, 243 I. ricinus were collected from 301 roe deer captures between 14 December and 28 February at the Bogesund study site during three subsequent years (2013/2014-2015/2016). We found attached ticks every third to every second examination (32%, 48% and 32% of the examinations, respectively). However, we collected only three I. ricinus females from 31 roe deer captures at the Grimsö study site between 17 December 2015 and 26 February 2016. At the Bogesund study site, based on 192 captures of previously examined deer, we collected 121 ticks, and ticks were found at 33%, 48% and 26% of the examinations during the respective winters. The probability of finding an attached tick on a roe deer at a temperature of -5 °C was > 8% ± 5 (SE), and that probability increased to almost 20% ± 7 (SE) if the air temperature increased to 5 °C. CONCLUSIONS: To the best of our knowledge, this is the first time that winter-active nymphs and female ticks have been documented to attach and feed on roe deer during winter (December to February) in Scandinavia. The main weather conditions regulating winter activity for females were temperature and precipitation, and the lowest estimated air temperature for finding an active tick was well below 5 °C. The behaviour of winter-active and blood-feeding ticks was documented over several winters and in two contrasting areas, implying that it is a common phenomenon that should be investigated more thoroughly, since it may have important consequences for the epidemiology of tick-borne pathogens.

A morphological investigation of the tongue of roe deer (Capreolus capreolus Linnaeus, 1758).

The morphological structure of the tongue and papillae that occur on it vary according to an animal's lifestyle, nutrition, and adaptation to various environmental conditions. This study aimed to reveal in detail the morphological, histological, and electron microscopic structure of the tongue of roe deer (Capreolus capreolus Linnaeus, 1758). In this study, nine roe tongues were used. The tongue consists of three parts: the apex, body, and root. When the dorsal surface of the tongue was examined in detail, five different papillae were observed: filiform, lenticular, conical, fungiform, and vallate. Filiform papillae differed in having secondary papillae according to their localization. The opening holes of taste buds were observed on the surface of the round and flat fungiform papillae. The free ends of the filiform papillae were more pointed and thinner than those of the other papillae, while the width of the lenticular papillae was thicker, the surface was flat, and the free ends were blunt. Triangular-shaped conical papillae were observed differently regarding the presence or absence of secondary papillae. The vallate papillae were caudolateral to the lingual torus. On the surface of the vallate papillae, circumferenced by a deep groove, were the opening holes of the taste buds and microridges. From this analysis, it appears to be characteristic of roe deer that mechanical function, filiform, and conical papillae contain secondary papillae; lenticular papillae, absent in many deer species, are found; and a prominent papillary groove surrounds all mechanical and gustatory papillae. RESEARCH HIGHLIGHTS: The lingual papillae of roe deer (Capreolus capreolus Linnaeus, 1758) were examined with this study in detail for the first time. Similarities and differences with ruminant species were determined.

No evidence of Mycobacterium tuberculosis complex infection in samples from cervids in various regions of Poland.

INTRODUCTION: Tuberculosis (TB) is a widespread disease known to affect livestock and wildlife, as well as humans. However, its incidence in wildlife remains poorly recognized on the global level. In Europe, the majority of TB cases have been confirmed in red deer, badgers and wild boar. OBJECTIVE: The aim of the study was to examine the occurrence of TB in Cervidae in Poland, in areas where TB has been detected in cattle and wildlife. MATERIAL AND METHODS: Head and thoracic lymph nodes were collected from a total of 76 free-living red deer (Cervus elaphus) and roe deer (Capreolus capreolus), from nine Polish provinces during a single hunting season (autumn - winter 2018-19). Samples were subjected to conventional microbiological procedures to isolate mycobacteria. RESULTS: No Mycobacteria was isolated in the material collected from red or roe deer. CONCLUSIONS: There is a need to continue monitoring the presence of TB in cattle and other animal species to ensure the protection of public health.

Seroprevalence of tick-borne encephalitis virus in wild and domestic animals in northern Germany.

Tick-borne encephalitis virus (TBEV) is a tick-transmitted flavivirus, which can infect humans and animals, sometimes even with a fatal outcome. Since many decades, TBEV is endemic in southern Germany, while only sporadic occurrence has been noted in northern parts of the country so far. Nevertheless, autochthonous human clinical cases are increasing in the federal state of Lower Saxony in north-western Germany, and several natural foci of TBEV transmission have recently been detected in this federal state. In order to shed more light on the current distribution of TBEV in Lower Saxony, the present study examined blood samples from wild and domestic animals for antibodies against TBEV. Overall, samples from 4,085 animals were tested by ELISA, including wild boar (N = 1,208), roe deer (N = 149), red deer (N = 61), fallow deer (N = 18), red foxes (N = 9), nutria (N = 9), raccoon dogs (N = 3), raccoons (N = 3), badgers (N = 1), European pine martens (N = 1), horses (N = 574), sheep (N = 266), goats (N = 67), dogs (N = 1,317) and cats (N = 399). Samples with an ELISA result of ≥60 Vienna units (VIEU)/ml were subjected to confirmatory serum neutralization tests (SNT). In total, 343 of 4,085 (8.4%) animals tested positive for anti-TBEV-IgG by ELISA, of which 60 samples were confirmed by SNT. Samples of 89 animals showed a cytotoxic effect in the SNT and were excluded from seroprevalence calculation, resulting in an overall seroprevalence of 1.5% (60/3,996). Seroprevalence was higher among wild animals (wild boar: 2.9% [34/1,190], roe deer: 2.7% [4/149], red deer: 1.7% [1/60], fallow deer: 5.6% [1/18]) than among domestic animals (dogs: 1.1% [15/1,317], horses: 0.8% [4/505], sheep: 0.4% [1/266]). No anti-TBEV-antibodies were detected in the other wild animal species as well as goats and cats. A notable clustering of positive samples was observed in districts where TBEV transmission foci have been described. Further clusters in other districts suggest the existence of so far undetected transmission foci, underlining the fact that both wild and domestic animals are useful sentinels for monitoring the spread of TBEV.

Cephenemyia stimulator (Diptera: Oestridae) myiasis in a roe deer (Capreolus capreolus) from Portugal.

Herein we present a nasopharyngeal myiasis case by Cephenemyia stimulator in a roe deer hunted in Trás-os-Montes (NE Portugal). Preliminary inspection showed one larva sorting through the nostrils and the examination of the nasopharyngeal cavity showed more than 15 larvae in the glottis and retropharyngeal recesses. Four larvae were collected and stored into 70% ethanol for morphological and molecular identification. Three of the larvae were identified as third instars and the other one as a prepupa of Cephenemyia stimulator, being the first confirmation of this species in roe deer from Portugal. C. stimulator is currently widely distributed in roe deer from central and northern Spain so, the transboundary natural dispersal of these cervids would explain the introduction of this myiasis in Portugal. Further studies are needed to monitor the spreading of this infection in the westernmost populations of European roe deer.

Hatchability of Fascioloides magna Eggs in Cervids.

The giant liver fluke (Fascioloides magna) is an invasive parasite found permanently in three foci in Europe. The fluke has an indirect life cycle involving a final and an intermediate host. The currently accepted terminology determines three types of final hosts: definitive, dead-end, and aberrant hosts. Recently, roe deer (Capreolus capreolus) has been classified as an aberrant host, which cannot contribute to the reproduction of F. magna. This study investigated the hatchability of F. magna eggs of red deer (Cervus elaphus) and roe deer origin to compare the suitability of the two host species for the maintenance of the parasite. The study was carried out on a newly invaded area, two years after the first reported observation of F. magna. The prevalence of the parasite proved to be 68.4% (CI95% 44.6-85.3%) in red deer and 36.7% (CI95% 24.8-50.0%) in roe deer. The difference between the two species was confirmed to be significant (p = 0.02). The mean intensity proved to be 10.0 (CI95% 4.9-22.6) and 7.59 (CI95% 2.7-24.2) in the red deer and the roe deer, respectively. The difference of the mean intensities did not prove to be significant (p = 0.72). Of the 70 observed pseudocysts, 67 originated from red deer and 3 from roe deer. Most of the pseudocysts contained two flukes, while a few pseudocysts contained one or three parasites. Egg production was observed in all three types of pseudocysts. We did not find more than three flukes in any pseudocyst. The apparent proportion of self-fertilisation in flukes without mating partners was 23.5% and 100% in red deer and roe deer, respectively. The survival of single-parent eggs was not confirmed to be worse than that of gregarious parents. The viability of offspring originating from roe and red deer differed significantly. Our findings suggest that F. magna adapted to the new populations of susceptible hosts rather than vice versa.