Identification and Molecular Characterization of Giant Liver Fluke (Fascioloides magna) Infection in European Fallow Deer (Dama dama) in Romania-First Report.

Fascioloidosis is a parasitic disease of primary wild and domestic ruminants, caused by giant liver fluke, Fascioloides magna. The definitive host of the liver fluke in its area of origin (North America) is the white-tailed deer (Odocoileus virginianus). In Europe, the red deer (Cervus elaphus) and European fallow deer (Dama dama) are definitive hosts and the most sensitive hosts to F. magna infection, on which the parasite exerts serious pathogenic effects. In this study, we analyzed fecal samples and livers of 72 D. dama from 11 hunting grounds in Arad County, Romania. Of the 72 fecal samples and livers from D. dama, trematodes of the genus Fascioloides were identified in four (5.56%). Sequencing revealed that the trematodes identified in the samples were similar to the sequence of F. magna (GenBank no. EF534992.1, DQ683545.1, KU232369.1). The sequence obtained from the molecular analysis has been deposited in GenBank® under accession number OQ689976.1. This study describes the first report of giant liver fluke (F. magna) infection in D. dama in Romania.

First Molecular Data of Gongylonema pulchrum (Rhabditida: Gongylonematidae) in European Fallow Deer Dama dama from Romania.

Due to its adaptive versatility to numerous types of habitats, extremely diverse both in terms of composition and specificity, developed in various areas of the Western Plains of Romania, the European fallow deer (Dama dama) is a species with high ecological plasticity. In this area, the D. dama interacts with other species of wild fauna but also with numerous domestic animals, an important aspect in terms of the sanitary-veterinary status of animal populations, as well as the existence of a potential risk of infection with various species of parasites that can cause the D. dama specimens to obtain certain diseases and even zoonoses. A total of 133 esophagi from D. dama have been examined for helminths. Of the 133 esophagus samples from D. dama, nematodes of the genus Gongylonema were identified in 25 (18.80%). Sequencing revealed that the nematode identified in the samples was 99% similar to the sequence of Gongylonema pulchrum (GenBank no. LC026018.1, LC388754.1, AB646061). The present research is the first report of the nematode G. pulchrum from D. dama in Romania.

First Identification and Molecular Characterization of Trichinella britovi (Nematoda: Trichinellidae) from the Pine Marten (Martes martes Linnaeus, 1758) in Romania.

Trichinella spp. are etiological zoonotic agents spread throughout the world and affect mammals, birds, and reptiles; they evolve via two cycles: domestic and sylvatic. Martes martes is a carnivorous nocturnal mammal from the family Mustelidae. In Romania, this host is widespread in all forests of the country. Martes martes has an extremely voracious appetite, feeding on fruit and also on a variety of small animals, including rodents such as mice and rats. The aim of this study was the identification and molecular characterization of Trichinella larvae isolated from the muscle tissue of Martes martes collected in different counties of Romania. The muscle samples were examined via artificial digestion, and the larvae were identified at the species level via multiplex PCR. The presence of larvae belonging to Trichinella britovi, a species frequently identified in wild carnivores in temperate zones, was observed. Although T. britovi has been already reported in several host species in Romania, this is the first time this species has been observed in a Martes martes specimen. This finding contributes to our knowledge about the host species involved in the maintenance of the Trichinella sylvatic cycle in Romania, and it confirms that this parasite is consistently present in the wild fauna of this country.

The First Identification of Trichinella britovi in the Raccoon Dog (Nyctereutes procyonoides) in Romania.

Trichinella spp. are nematodes distributed throughout the world that affect an impressive number of host animals (mammals, birds, and reptiles) involved in the evolution of two cycles, the domestic and the sylvatic. The raccoon dog (Nyctereutes procyonoides) is an omnivorous mammal with great ecological plasticity. The expansion of the raccoon dog in Europe is associated with the risk of the introduction and spread of different pathogens, especially zoonotic ones (Trichinella, Echinococcus). Currently, the raccoon dog's range in Romania is limited to the Danube Delta area, the Lower Danube Meadow, and the Prut Meadow. The aim of this study was to examine the presence of Trichinella larvae isolated from the muscles of raccoon dog from six hunting funds of Giurgeni, Ialomița County, Romania. The muscle samples were examined via artificial digestion, and the obtained larvae were processed via multiplex PCR. The PCR-amplified ESV and ITS1 DNA fragments were then sequenced for species confirmation. The species Trichinella britovi, which is the most common species identified in wild carnivores in temperate zones, was confirmed. Although T. britovi has been reported in several host animals in Romania, this case report confirms its presence in the raccoon dog for the first time.

First Morphological and Molecular Identification of Demodex injai in Golden Jackal (Canis aureus Linnaeus, 1758) in Romania.

Demodicosis is one of the most important external parasitic diseases found in carnivores. Three species of the Demodex mite inhabit the skin of dogs and related species, D. canis being the most prevalent. This paper describes the first case of infestation with D. injai in a golden jackal in Romania. An emaciated golden jackal female body found in Timiș County, western Romania, was examined at Parasitology Department of Faculty of Veterinary Medicine, Timișoara. The gross lesions were present on different regions of the body: feet, tail, axillary and inguinal areas, and skin folds as well, consisting of erythema, extensive severe alopecia with lichenification, seborrhea, and scaling. In order to establish diagnosis, microscopic examination of skin scrapes, trichogram (hair plucking), acetate tape test (impression), fungal culture, and PCR were performed. Both microscopic measurements and PCR analysis have confirmed the presence of D. injai.

First Identification of Trichinella pseudospiralis in a Golden Jackal (Canis aureus) in Romania.

Trichinella spp. are etiological zoonotic agents that spread throughout the world and affect mammals, birds, and reptiles. Within this genus, Trichinella pseudospiralis is the only recognized non-encapsulated species known to infect mammals and birds. This species has been reported in the majority of European countries, and the real epidemiological scenario of this species remains to be defined because its detection in mammals is much lower than that of the capsulated species. The aim of this study was to examine the presence of Trichinella larvae isolated from the muscles of a jackal from the hunting fund of 36 Murfatlar, Constanta County, Romania. The muscle samples were examined by artificial digestion, and the larvae were identified at the species level by multiplex PCR. The presence of larvae belonging to T. pseudospiralis, a species more frequently reported in carnivorous birds, was observed. This study describes the first identification of T. pseudospiralis in a jackal. The results suggest that there is an urgent need to investigate which species of mammals and/or birds act as reservoirs for this zoonotic nematode in Romania.