The red fox (Vulpes vulpes) is widely distributed in the world; in central Europe, it is the most numerous wild species of the family Canidae. It can play the role of a definitive host for many intestinal parasites, including zoonotic helminths. Poland, with its geographical location (central Europe), is an interesting area for parasitological investigations of this species. The aim of this study was to evaluate and compare the prevalence of intestinal helminths in red foxes in different regions of Poland. Intestines of 473 red foxes from four different regions were examined using the sedimentation and counting technique (SCT). In addition, 344 samples of faeces were examined using flotation. Overall, intestinal helminths were found in 98.9% of red foxes. The average prevalence of detected parasites was as follows: Mesocestoides spp. (84.1%); hookworms (67.9%); Alaria alata (61.5%); Toxocara/Toxascaris (49.5%); Taenia spp. (42.5%); Echinococcus multilocularis (25.6%); and Trichuris vulpis (2.3%). The prevalence of the majority of parasite species was similar in each region. Significant differences between regions were observed in the case of E. multilocularis: a low prevalence in the south-western and northern regions (0% and 0.9%, respectively) and a high prevalence in the south-east and northeast (39.3% and 42.7%, respectively). In the case of A. alata, important differences were found between northern (96.5% and 93.7% in northern and northeast regions, respectively) and southern regions (15.2% and 24.7% for south-western and south-east regions, respectively). The percentage of positive samples obtained with coproscopic examination (except for Trichuris) was significantly lower than that obtained with SCT. Analysis of the prevalence estimated in individual regions with the use of both methods (flotation and SCT) showed a high correlation for all parasite species (except for Mesocestoides spp.). The flotation method also allowed us to detect the eggs of the lung nematode Eucoleus aerophilus (syn. Capillaria aerophila) (76.2% of positive foxes). This study showed a very high percentage of red foxes infected with intestinal helminths in different parts of Poland. Depending on the location, some differences were observed regarding the prevalence of dangerous zoonotic parasites, which should be considered in the assessment of infection risk for humans.

Background: Eucoleus aerophilus (syn. Capillaria aerophila) is a nematode with a worldwide geographical distribution. It causes a disease called lung capillariosis by affecting the respiratory tract of wild and domestic animals, and has also occasionally been described in humans. Despite steady increases in knowledge of the morphology of this neglected parasite, many aspects are still poorly understood. Epidemiological data regarding, for example, geographic distribution, range of hosts, clinical relevance and the actual zoonotic potential of this nematode are scarce and incomplete. Methods: This article is a systematic review based on the screening of three databases (PubMed, Web of Science and Science Direct) to identify eligible studies published from 1973 to the end of 2022. Results: From a total of 606 studies describing the occurrence of E. aerophilus, 141 articles from 38 countries worldwide were included in this meta-analysis, all of which presented results obtained mainly with flotation and necropsy. Due to the occurrence of E. aerophilus in many different species and different matrices (lungs and faeces), we decided to conduct the meta-analysis separately for each species with a given matrix. This systematic review confirmed the status of the Red fox as the main reservoir and main transmitter of E. aerophilus (average prevalence of 43% in faeces and 49% in lungs) and provided evidence of a higher prevalence of E. aerophilus in wild animals in comparison to domestic animals, such as dogs (3% in faeces) and cats (2% in faeces and 8% in lungs). Previous studies have investigated many host-related factors (age, sex, environmental/living conditions) in relation to the prevalence of E. aerophilus, but they show wide variations and no simple relationship has been demonstrates. Furthermore, mixed infections with other pulmonary nematodes, such as Crenosoma vulpis and/or Angiostrongylus vasorum, are reported very frequently, which greatly complicates the diagnosis. Conclusions: This systematic review focused on identifying data gaps and promoting future research directions in this area. To the best of our knowledge, this is the first systematic review that evaluates and summarizes existing knowledge on the occurrence and prevalence of E. aerophilus in wild and domestic animals originating from different geographical locations worldwide. Graphical

The aim of this study was to provide molecular characterization, together with phylogenetic analysis, of Taenia pisiformis cysts isolated from rabbit. On the basis of morphological features and molecular analysis, the cysticerci were identified as T.pisiformis metacestodes. PCR was performed with three different protocols to obtain partial sequences of 12S ribosomal RNA (12S rRNA), NADH dehydrogenase subunit 1 (nad1), and cytochrome oxidase subunit 1 (cox1) of Taenia spp. The products from the PCRs were sequenced. Interpretation of the sequencing results of the obtained amplicons, by comparing them with the GenBank database, proved that the causative agent, in this case, was T. pisiformis. The phylogenetic analysis of the received sequences identified a new haplotype. The received data can be used to supplement the species description. To our knowledge, this is the first molecular confirmation of T. pisiformis metacestodes infection in the rabbit, in Poland.

Introduction: The aim of the study was to determine the genetic diversity of Echinococcus multilocularis in pigs in highly endemic areas in Poland, as well as to attempt to confirm the occurrence and geographical distribution of haplotypes characteristic for these areas, which were previously described on the basis of examination of adult tapeworms isolated from foxes. Material and Methods: Twenty samples of E. multilocularis larval forms were obtained from pigs’ livers in four provinces of Poland. Genetic analyses were conducted on sequences of two mitochondrial genes: cox1 and nad2. Results: Seven haplotypes were found for the cox1 gene (OQ874673–OQ874679) and four haplotypes for nad2 (OQ884981–OQ884984). They corresponded to the haplotypes described earlier in foxes in Poland (some of them differing only in one nucleotide). The analysis showed the presence of the Asian-like haplotype in both the cox1 and nad2 genes. The remaining haplotypes were grouped in the European clade. The geographical distribution of haplotypes identified in the pig samples was noticed to bear a similarity to the distribution of haplotypes previously isolated from foxes in the same regions. Conclusion: The characteristic geographical distribution of E. multilocularis haplotypes in Central Europe (including the presence of the Asian-like haplotype) previously described in the population of definitive hosts (foxes) has now been confirmed by the analysis of samples from non-specific intermediate hosts (pigs).

The aim of this study is to confirm the presence and molecular identification of Echinococcus tapeworms in wolves from south-eastern Poland. An investigation was carried out on the intestines of 13 wolves from south-eastern Poland. The small intestines were divided into three equal segments. Each segment was separately examined using the sedimentation and counting technique (SCT). The detected Echinococcus tapeworms were isolated and identified by PCRs and sequencing (nad1 and cox1 genes). Additionally, DNA isolated from the feces of wolves positive for Echinococcus tapeworms was examined with two diagnostic PCRs. The intestines of one wolf were positive for E. granulosus s.l. when assessed by SCT; the intestine was from a six-year-old male wolf killed in a communication accident. We detected 61 adult tapeworms: 42 in the anterior, 14 in the middle, and 5 in the posterior parts of the small intestine. The PCRs conducted for cox1 and nad1 produced specific products. A sequence comparison with the GenBank database showed similarity to the deposited E. ortleppi (G5) sequences. An analysis of the available phylogenetic sequences showed very little variation within the species of E. ortleppi (G5), and identity ranged from 99.10% to 100.00% in the case of cox1 and from 99.04% to 100.00% in the case of nad1. One of the two diagnostic PCRs used and performed on the feces of Echinococcus-positive animals showed product specific for E. granulosus. This study showed the presence of adult E. ortleppi tapeworms in wolves for the first time.

Introduction: The aim of this study was to estimate the occurrence of Echinococcus spp. and other helminth infections in grey wolves in south-eastern Poland. Material and Methods: Overall, 74 samples of wolf faeces were examined with a multiplex PCR and a system of real-time quantitative PCR methods to detect and identify Echinococcus spp. The faeces were additionally examined microscopically. Also, 20 samples of wolf intestines were examined with a sedimentation and counting technique (SCT). Results: Echinococcus multilocularis DNA was detected in 6.8% and E. granulosus s.l . (identified as E. ortleppi ) in 4.1% of faeces samples. Taenia spp. DNA was found in 43.2% and Mesocestoides in 4.1%. Examination of the intestines by SCT showed E. multilocularis worms in 10%, E. granulosus s.l . ( E. ortleppi ) in10%, Taenia spp. in 100%, hookworms in 30%, Alaria alata in 20%, Mesocestoides sp. in 10%, Trichuris vulpis in 15%, Molineus sp. in 5% and Euryhelmis sp. in 5%. By coproscopy, Capillariidae eggs were found in 59% of faeces samples. Genetic analysis of E. multilocularis worms showed the presence of two European haplotypes previously described in Poland in red foxes and pigs. Sequences of nad1 obtained from E. ortleppi worms shared full identity with a sequence from a human case in Poland. Conclusion: The study showed the presence of E. multilocularis in wolves for the first time in Poland and confirmed our earlier observations on E. ortleppi . This double threat from Echinococcus in this wolf population should be taken into account when assessing the epidemiological risk. The study enriched the knowledge of other helminths found in wolves, also those ( Euryhelmis ) that were recorded for the first time in this species.