Parasites are among the most common pathogens recorded in dogs. High prevalence rates are usually reported in stray and shelter dogs, as these animals are less likely to be tested and treated. In this study a survey was carried out on intestinal and lung parasites of stray dogs at the moment of the admission in a shelter in Central Italy. In the period June 2014-June 2015, 262 individual faecal samples were examined. Twelve parasitic taxa were detected. Helminths were more prevalent than protozoa (61.8% and 25.6%, respectively). Ancylostomatidae showed the higher prevalence (40.5%), followed by Giardia duodenalis (21.4%), Toxocara canis (20.6%) and Trichuris vulpis (17.6%). Angiostrongylus vasorum was the most prevalent lungworm (12.6%), Crenosoma vulpis being recorded just in one dog. G. duodenalis had a high prevalence, probably because of the diagnostic techniques adopted, actually the most sensitive available for its detection. Nevertheless, the zoonotic potential of this parasite in dogs appears reduced, as all the isolates were identified as C (24.0%) and D (76.0%) dog specific assemblages. Taeniidae constituted a high potential zoonotic risk, as from the eggs it is impossible to exclude they were Echinococcus granulosus, the most relevant zoonotic parasite in Europe. The present study underline the importance of stray dogs control, aimed at preventing or minimizing parasitic spread and zoonotic transmission.

Free-living amoebae of the genus Acanthamoeba are worldwide present in natural and artificial environments, and are also clinically important, as causative agents of diseases in humans and other animals. Acanthamoeba comprises several species, historically assigned to one of the three groups based on their cyst morphology, but presently recognized as at least 20 genotypes (T1-T20) on the basis of their nuclear 18S ribosomal RNA (rRNA) gene (18S rDNA) sequences. While strain identification may usually be achieved targeting short (<500 bp) 18S ribosomal DNA (rDNA) fragments, the use of full-length gene sequences (>2200 bp) is necessary for correct genotype description and reliable molecular phylogenetic inference. The genotype T15, corresponding to Acanthamoeba jacobsi, is the only genotype described on the basis of partial sequences (~1500 bp). While this feature does not prevent the correct identification of the strains, having only partial sequences renders the genotype T15 not completely defined and may furthermore affect its position in the Acanthamoeba molecular tree. Here, we complete this gap, by obtaining full-length 18S rDNA sequences from eight A. jacobsi strains, genotype T15. Morphologies and physiological features of isolated strains are reported. Molecular phylogeny based on full 18S rDNA confirms some previous suggestions for a genetic link between T15 and T13, T16, and T19, with T19 as sister-group to T15.

Free-living amoebae (FLA) are protozoa ubiquitous in Nature, isolated from a variety of environments worldwide. In addition to their natural distribution, some species have been found to be pathogenic to humans. In the present study a survey was conducted in order to evaluate the presence and to characterize at molecular level the isolates of amoebic organisms collected from different water sources in Italy. A total of 160 water samples were analyzed by culture and microscopic examination. FLA were found in 46 (28.7%) of the investigated water samples. Groundwater, well waters, and ornamental fountain waters were the sources with higher prevalence rates (85.7%, 50.0%, and 45.9%, respectively). Identification of FLA species/genotypes, based on the 18S rDNA regions, allowed to identify 18 (39.1%) Acanthamoeba isolates (genotypes T4 and T15) and 21 (45.6%) Vermamoeba vermiformis isolates. Other FLA species, including Vahlkampfia sp. and Naegleria spp., previously reported in Italy, were not recovered. The occurrence of potentially pathogenic free-living amoebae in habitats related to human population, as reported in the present study, supports the relevance of FLA as a potential health threat to humans.

Swimmer's itch is caused by the penetration of free-swimming larvae of trematodes of the family Schistosomatidae in human skin. It is usually reported in people engaged in recreational water activities in freshwater bodies and in most of cases, it is provoked by bird schistosomes of the genus Trichobilharzia. In the summer 2017, many cases of dermatitis were recorded in people bathing in the waters of the Albano Lake (Rome, Italy) and a parasitological investigation was carried out in order to ascertain the causative agent of these cases. Snails of the family Lymnaeidae, natural intermediate hosts of bird schistosomes, were collected from lake shallow waters to detect the presence of trematodes of the genus Trichobilharzia. Pools of maximum 10 snails were placed in Petri dishes, and cercarial emergence was stimulated exposing snails to strong artificial light intensity at 25°C. Three hundred and thirty-seven snails were collected and screened for the shedding of cercariae. Furcocercariae of the family Schistosomatidae, with a morphology overlapping that of the genus Trichobilharzia, were detected in seven Petri dishes. Assuming that in each positive Petri dish just one snail was shedding furcocercariae, the minimum infectious rate was 2.1%. Molecular analysis of furcocercariae allowed ascribing them to the species Trichobilharzia franki.Snails of the species Radix auricularia were identified as intermediate hosts of the parasite. This is the second record of T. franki causing cercarial dermatitis in Central Italy, the third in Italy. The 2017 was in Italy exceptionally warm and dry. Trematodes are sensitive to changes in temperature, being cercarial production and emission rates temperature dependent. Small increases in water temperature would speed up parasite development and transmission, leading to a manifold increase in cercarial emergence. Moreover, high temperatures raise chances to acquire the infection, due to increased time spent in water by people.

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Giardia duodenalis is the most common intestinal protozoan in humans and animals worldwide, including eight morphologically identical assemblages, infecting pets, livestock, wildlife and human beings. Assemblages A and B are those with the higher zoonotic potential, and they have been detected in several mammals other than humans; the others (C to H) show a higher host specificity. Cats can harbour both the specific Assemblage F and the zoonotic ones A and B. Several studies have been carried out on G. duodenalis genotypes in cats; however, the role of this species in the epidemiology of giardiasis is still poorly understood. In this scenario, the present study carried out the detection and genetic characterization at sub-assemblage level of G. duodenalis from colony stray cats in central Italy. In the period 2018-2019, 133 cat faecal samples were analysed for the presence of G. duodenalis cysts by a direct immunofluorescence assay. Positive samples were subsequently subjected to molecular analyses for assemblage/sub-assemblage identification. Forty-seven samples (35.3%) were positive for G. duodenalis cysts by immunofluorescence. G. duodenalis DNA was amplified at SSU-rDNA locus from 39 isolates: 37 were positive for zoonotic Assemblage A and 2 showed a mixed infection (A + B). Positive results for the β-giardin gene were achieved for 25 isolates. Sequence analysis revealed 16 isolates belonging to Sub-assemblage AII and 8 to Sub-assemblage AIII. One isolate resulted as ambiguous AI/AIII. Large sequence variability at the sub-assemblage level was detected, with several double peaks and mutations, making complex a proper isolate allocation. When compared with previous studies, the 35.3% prevalence of G. duodenalis in cats reported in the present article was surprisingly high. Moreover, all positive cats resulted to be infected with zoonotic assemblages/sub-assemblages, thus indicating stray cats as a possible source of human giardiasis and highlighting the sanitary relevance of cat colonies in the study area.

Background: Canine primary chronic enteropathy (CE) includes a heterogeneous group of diseases characterized by chronic gastrointestinal signs.Aim: This study evaluated the occurrence of Giardia duodenalis infection in primary CE-affected dogs.Methods: Forty-seven CE-affected dogs of different age and sex were enrolled in the study. For each dog, frequency of defecation, fecal consistency, and eventual fecal abnormalities were evaluated. A clinical scoring index of CE severity (clinical chronic enteropathy activity index) was also assessed, and the type of enteropathy was retrospectively classified. For parasitological analysis, fresh fecal samples collected from each dog were examined by fresh and Lugol stained smears, flotation test, and a rapid immunoassay. Giardia duodenalis genotypes were identified by molecularanalysis. Differences of clinical parameters between G. duodenalis positive and G. duodenalis negative dogs were statistically evaluated.Results: Among the CE canine patients, 16 out of 47 (34%) dogs were found positive for G. duodenalis and assemblages C and D were identified. No statistical differences emerged according to the types of CE between G. duodenalispositive and G. duodenalis-negative dog groups. The clinical index of CE severity was indicative of significant less severe clinical forms in G. duodenalis-positive dogs (p = 0.037).Conclusion: Results here obtained shows how G. duodenalis may be present in primary CE-affected dogs and further investigations are needed to clarify the real significance of mild clinical presentation in G. duodenalis-positive dogs affected by CE. Keywords: Canine, Clinical forms, Genotypes, Giardiasis, Primary chronic enteropathy.

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