Global and local drivers of Echinococcus multilocularis infection in the western Balkan region.

The cestode, Echinococcus multilocularis, is one of the most threatening parasitic challenges in the European Union. Despite the warming climate, the parasite intensively spread in Europe's colder and warmer regions. Little is known about the expansion of E. multilocularis in the Balkan region. Ordinary least squares, geographically weighted and multi-scale geographically weighted regressions were used to detect global and local drivers that influenced the prevalence in red foxes and golden jackals in the southwestern part of Hungary. Based on the study of 391 animals, the overall prevalence exceeded 18% (in fox 15.2%, in jackal 21.1%). The regression models revealed that the wetland had a global effect (ß = 0.391, p = 0.006). In contrast, on the local scale, the mean annual precipitation (ß = 0.285, p = 0.008) and the precipitation seasonality (ß = - 0.211, p = 0.014) had statistically significant effects on the infection level. The geospatial models suggested that microclimatic effects might compensate for the disadvantages of a warmer Mediterranean climate. This study calls attention to fine-scale analysis and locally acting environmental factors, which can delay the expected epidemic fade-out. The findings of our study are suggested to consider in surveillance strategies.

Evaluation of a Phytogenic Feed Supplement Containing Carvacrol and Limonene on Sheep Performance and Parasitological Status on a Hungarian Milking Sheep Farm.

There is currently worldwide interest in phytogenic feed supplements (PFSs) because they can lead to improved animal production. The scope of the present study was to observe the impact of a feed supplement containing carvacrol (CAR) and limonene (LIM) on the performance and parasitological status of sheep. The feed supplement decreased the plasma levels of ß-hydroxybutyrate (p < 0.001), triglycerides (p = 0.014), nonesterified fatty acids (p = 0.021), and fructosamine (p = 0.002) in lactating ewes after 42 days of supplementation, while the average live weight (p = 0.002) and average daily weight gain (p = 0.001) of their twin suckling lambs increased significantly by the end of the study. In another experiment, fattening lambs fed the same supplement showed a decrease in fecal egg number of gastrointestinal nematodes (p = 0.02) but no differences in live weight, average daily gain, or mean number of Haemonchus contortus nematodes in the abomasum. The results highlighted that the inclusion of carvacrol and limonene in the feed of lactating ewes effectively increased the weight gain of the suckling lambs, presumably due to the ewes' improved energy, but further studies are needed to elucidate the effects of carvacrol and limonene against gastrointestinal parasites.

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.

Survey of wild boar hunter interactions with pig farming in central Europe.

African swine fever (ASF) is a fatal animal disease without zoonotic potential but greatly impacts human well-being, especially in the most vulnerable human communities. In Europe, ASF concerns mostly the wildlife domain of health. The main vector of the disease is confirmed to be the wild boar, though long-distance jumps of the infection are due to anthropogenic effects. This study aimed to evaluate the potential role of hunting assistant personnel (beaters and carcass handlers) in ASF spread in Hungary. Based on a personal interview survey, we attempted to identify the epidemiological risk caused by hunting activities and the hunting personnel. The interviews with 58 hunting workers confirmed that an extent backyard pig sector (13 pig farmers) and pork production system (31 pork producers) existed within the study region out of the authorities' sight. Two pig farmers did not wear special working clothes for pig caring, seven pork producers disposed of slaughter offal in the settlements periphery, and six persons regularly contacted distant pig farms. The revealed knowledge, attitude, and practice of the questioned pig farmers suggested that this sector would be very vulnerable in an epidemic situation; moreover, backyard farms would cause a great risk for wildboar populations. Considering that the study region is the third poorest region of Hungary, these findings called attention to the high epidemiologic risk of socioeconomic inequality between different regions within the European Union.

The golden jackal (Canis aureus) and the African swine fever pandemic: Its role is controversial but not negligible (a diet analysis study).

BACKGROUND: In Europe, the African swine fever (ASF) pandemic mostly affects the environmental domain of health, which is a strongly human-impacted ecosystem. However, the current control strategies focus solely on the wild boar and tend to disregard other epidemiologically relevant elements of the ecosystem. OBJECTIVES: This study investigated the potential impact of the golden jackal on the surveillance effort and disease transmission. METHODS: For this reason, the authors analysed the content of 277 stomachs of this canid species within its westernmost inhabitant population, in order to determine the amount of suid remains, disposed. RESULTS: The findings confirmed that in a densely populated wild boar habitat, the main diet component of jackals was wild boar all the year round. The jackals disposed of 0.3-0.6 kg/km2 /day offals that potentially contained suid remains. On the other hand, the scavenging activity removed the most important target objects on which the passive surveillance of ASF should be based. CONCLUSIONS: This study cannot determine whether canid scavengers positively or negatively influence the control efforts; however, the impact of the jackal should not be disregarded. The results warn the necessity of a multidisciplinary approach to complex epidemiological situations within different ecosystems.

Habitat Characteristics as Potential Drivers of the Angiostrongylus daskalovi Infection in European Badger (Meles meles) Populations.

From 2016 to 2020, an investigation was carried out to identify the rate of Angiostrongylus spp. infections in European badgers in Hungary. During the study, the hearts and lungs of 50 animals were dissected in order to collect adult worms, the morphometrical characteristics of which were used for species identification. PCR amplification and an 18S rDNA-sequencing analysis were also carried out. Global and local spatial autocorrelation methods were used to detect high-rated and low-rated infected animal clusters. We conducted a binary logistic regression analysis along with hierarchical agglomerative clustering to determine the relation between selected biotic and abiotic variables, and the prevalence of an A. daskalovi infection. We found a high prevalence (72%) and moderate mean intensity (14.1) of Angiostrongylus sp. infection. Morphology and sequencing revealed that all animals were infected by A. daskalovi. The results of both spatial autocorrelations suggested that the spatial distribution of infected badgers was more spatially clustered than random. The results of an analysis of the correlation between habitat characteristics and infection showed that the infected animals could be associated with dry and open landscape habitats without extended and connected canopy. It is suggested that the territorial behaviour of badgers and the landscape-directed aggregation of potential intermediate hosts might be the drivers of an A. daskalovi infection.

The occurrence of Echinococcus spp. in golden jackal (Canis aureus) in southwestern Hungary: Should we need to rethink its expansion?

Alveolar echinococcosis and cystic echinococcosis are severe zoonotic diseases caused by Echinococcus multilocularis and Echinococcus granulosus s.l. in Europe. To present knowledge, in the European continent, the most important definitive hosts of these parasites belong to the Canidae family. The golden jackal as an opportunistic mesopredator frequently preys on rodents including arvicolids and other easily available food resources, such as viscera and other carrion. By these reasons, the golden jackal can promote the maintenance of both Echinococcus multilocularis and Echinococcus granulosus s.l. Our investigation was conducted in the southwestern part of Hungary where one of the densest golden jackal populations exists. We examined altogether 173 golden jackal small intestines to determine the presence of Echinococcus multilocularis and Echinococcus granulosus s.l. After the molecular diagnostic procedure, we found 27 Echinococcus multilocularis-positive (prevalence: 15.6%; mean intensity: 664 worms) and three Echinococcus granulosus s.l. infected hosts (prevalence: 1.7%; mean intensity: 554.3 worms). We suggest the invasion of the golden jackal in Europe can enhance the spread of both Echinococcus multilocularis and Echinococcus granulosus s.l. This novel epidemiological situation can influence the geographical distribution of these helminths and the characteristics of their endemic in different host species, as well as in humans.

Establishment of Fascioloides magna in a new region of Hungary: case report.

During the monitoring of red deer (N = 124) and fallow deer (N = 13) populations in four neighbouring areas, the presence of Fascioloides magna was confirmed in southwestern Hungary. The prevalence and the mean intensity of the infection within the host populations ranged between 0 and 100% and 0-36.3, respectively. The determined prevalences are similar to that observed earlier in other European natural foci. The authors hypothesise that the appearance of F. magna in this region should have been a partly natural- and partly human-influenced process.

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.

Benzimidazole resistance in Haemonchus contortus recovered from farmed red deer.

Thirty Haemonchus contortus male worms were collected from farmed red deer yearlings in order to determine whether routine administration of albendazole for a long-term period (17 years) could select anthelmintic resistance. PCR-RFLP method based on single-nucleotide polymorphism of codon 200 in isotype 1 ß-tubulin gene (Phe200Tyr) was applied. The results showed a significant frequency of either the resistant allele (85 %) or the homozygous resistant genotype (70 %). By chi-square test, Hardy-Weinberg equilibrium of the population was accepted (p = 0.334, power of test 0.01). True prevalence of the resistant genotype (RR) was estimated to be 46.5-87.2 % (confidence interval 95 %) calculated by Sterne's exact method. These results confirmed that long-term use of benzimidazoles could change the relative allele frequency of genes associated with drug resistance and may cause a large-scale spread of the resistant allele. To our knowledge, this study supported benzimidazole resistance in Haemonchus contortus in red deer for the first time.

Molecular analysis and MIRU-VNTR typing of Mycobacterium avium subsp. avium, 'hominissuis' and silvaticum strains of veterinary origin.

Besides Mycobacterium avium subsp. paratuberculosis (MAP), M. avium subsp. avium (MAA), M. avium subsp. silvaticum (MAS), and 'M. avium subsp. hominissuis' (MAH) are equally important members of M. avium complex, with worldwide distribution and zoonotic potential. Genotypic discrimination is a prerequisite to epidemiological studies which can facilitate disease prevention through revealing infection sources and transmission routes. The primary aim of this study was to identify the genetic diversity within 135 MAA, 62 MAS, and 84 MAH strains isolated from wild and domestic mammals, reptiles and birds. Strains were tested for the presence of large sequence polymorphism LSP(A)17 and were submitted to Mycobacterial interspersed repetitive units-variable-number tandem repeat (MIRU-VNTR) analysis at 8 loci, including MIRU1, 2, 3, and 4, VNTR25, 32, and 259, and MATR9. In 12 strains hsp65 sequence code type was also determined. LSP(A)17 was present only in 19.9% of the strains. All LSP(A)17 positive strains belonged to subspecies MAH. The discriminatory power of the MIRU-VNTR loci set used reached 0.9228. Altogether 54 different genotypes were detected. Within MAH, MAA, and MAS strains 33, 16, and 5 different genotypes were observed. The described genotypes were not restricted to geographic regions or host species, but proved to be subspecies specific. Our knowledge about MAS is limited due to isolation and identification difficulties. This is the first study including a large number of MAS field strains. Our results demonstrate the high diversity of MAH and MAA strains and the relative uniformity of MAS strains.

Molecular identification of Mycobacterium avium subsp. silvaticum by duplex high-resolution melt analysis and subspecies-specific real-time PCR.

Accurate identification of mycobacterial species and subspecies is essential to evaluate their significance and to perform epidemiological studies. The subspecies of Mycobacterium avium have different attributes but coincide in their zoonotic potential. Our knowledge about M. avium subsp. silvaticum is limited, since its identification is uncertain. Mycobacterium avium subsp. avium and M. avium subsp. silvaticum can be discriminated from each other based only on phenotypic characteristics, as they have almost identical genome sequences. Here we describe the development of a diagnostic method which enables the molecular identification of M. avium subsp. silvaticum and discrimination from M. avium subsp. avium based on genomic differences in a duplex high-resolution melt and M. avium subsp. silvaticum-specific mismatch real-time PCR. The developed assay was tested on reference strains and 199 field isolates, which were analyzed by phenotypic methods previously. This assay not only identified all 63 M. avium subsp. silvaticum and 138 M. avium subsp. avium strains correctly but also enabled the detection of mixed M. avium subsp. avium-M. avium subsp. silvaticum cultures. This is the first time that such a large panel of strains has been analyzed, and we also report the first isolation of M. avium subsp. silvaticum from red fox, red deer, wild boar, cattle, and badger. This assay is reliable, rapid, simple, inexpensive, and robust. It eliminates the long-existing problem of ambiguous phenotypic identification and opens up the possibility for detailed and comprehensive strain studies.

Wild boar density drives Metastrongylus infection in earthworm.

Larvae of Metastrongylus spp. lungworms infect wild boar (Sus scrofa) definitive hosts through earthworms (Lumbricidae). We compared the abundance and Metastrongylus spp. larval infection measures of earthworms between two areas (both in Zselic, Hungary, 2012) characterized by markedly different wild boar population densities. Estimated wild boar density was 0.03 animal/ha in free range area and 1.03 animal/ha in enclosure. The mean abundance of earthworm populations (mostly Allolobophora, Aporrectodea, and Lumbricus spp.) was assessed by analysing 140-140 soil samples. The assesment of Metastrongylus spp. larval infection measures was based on cca 100-100 earthworms derived from the two areas. The abundance of earthworms and their Metastrongylus spp. larval infection measures (prevalence and mean intensity) were significantly lower in the free range than in the enclosure. Furthermore, using a finer scale within the enclosure, we compared wild boar feeding sites (n = 30) to other sites (n = 75). Earthworm populations were significantly more abundant and carried significantly more prevalent and more abundant larval lungworm infections at the former sites. These results suggest that high wild boar density and forage supplementation in enclosures increase both the abundance and the larval Metastrongylus infections of earthworms.