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The Mediterranean Sea is recognized as a marine biodiversity hotspot. This enclosed basin is facing several anthropogenic-driven threats, such as seawater warming, pollution, overfishing, bycatch, intense maritime transport and invasion by alien species. The present review focuses on the diversity and ecology of specific marine trophically transmitted helminth endoparasites (TTHs) of the Mediterranean ecosystems, aiming to elucidate their potential effectiveness as ‘sentinels’ of anthropogenic disturbances in the marine environment. The chosen TTHs comprise cestodes and nematodes sharing complex life cycles, involving organisms from coastal and marine mid/upper-trophic levels as definitive hosts. Anthropogenic disturbances directly impacting the free-living stages of the parasites and their host population demographies can significantly alter the distribution, infection levels and intraspecific genetic variability of these TTHs. Estimating these parameters in TTHs can provide valuable information to assess the stability of marine trophic food webs. Changes in the distribution of particular TTHs species can also serve as indicators of sea temperature variations in the Mediterranean Sea, as well as the bioaccumulation of pollutants. The contribution of the chosen TTHs to monitor anthropogenic-driven changes in the Mediterranean Sea, using their measurable attributes at both spatial and temporal scales, is proposed.

Dirofilariasis is a zoonosis caused by nematodes of the genus Dirofilaria.Dirofilaria immitis is cosmopolitan as regards its distribution in animals, being responsible for human pulmonary dirofilariasis in the New World. However, human infections by Dirofilaria immitis are exceptional in Europe, and the previously reported Italian cases of pulmonary dirofilariasis were due to Dirofilaria repens. We performed a systematic literature review of the Italian cases of human dirofilariasis due to Dirofilariaimmitis according to the PRISMA guidelines. We also report the first autochthonous case of human pulmonary dirofilariasis due to Dirofilariaimmitis, confirmed by polymerase chain reaction analysis. The patient was a 60-year-old man who lived in the Po river valley and had never traveled abroad; on histological examination, the 2-cm nodule found in his right upper lung was an infarct due to a parasitic thrombotic lesion. Only one other autochthonous (but conjunctival) case due to Dirofilariaimmitis (molecularly confirmed) was previously found in the same geographic area. Climatic changes, the increasing movements of animal reservoirs and vectors, and new competent carriers have expanded the geographic distribution of the Dirofilaria species, increasing the risk of human infections. Our report demonstrates that at least some pulmonary Italian cases of human dirofilariasis are due to Dirofilaria immitis, as in the New World.

The family Acuariidae is a speciose group of parasitic nematodes, infecting mostly birds as definitive hosts. This study focused on the characterization of two species of acuariids, collected in two different species of piscivorous birds, the European great cormorant Phalacrocorax carbo sinensis from Italy, and the pygmy cormorant Microcarbo pygmaeus from Israel. Parasites were analyzed using light and scanning electron microscopy and by amplification and sequencing of the 28S rDNA. The results of morphological and molecular analyses showed that Ph. carbo sinensis was infected by the acuariid Syncuaria squamata (12 females) and Cosmocephalus obvelatus (1 female), whereas M. pygmaeus was infected by C. obvelatus (2 males, 12 females). The present results provide new data on the distribution of acuariid parasites of piscivorous birds, the first report of Acuariidae in Israel, and the first molecular data on S. squamata and C. obvelatus , which will be useful in future epidemiological and phylogenetic studies of these widely distributed, but less molecularly studied parasites.

The distribution of parasites is shaped by a variety of factors, among which are the migratory movements of their hosts. Israel has a unique position to migratory routes of several bird species leaving Europe to winter in Africa, however, detailed studies on the parasite fauna of birds from this area are scarce. Our study investigates occurrence and distribution of sibling species among Contracaecum rudolphii complex in Phalacrocorax carbo sinensis from Italy and Israel, to acquire further information on the geographical range of these species to gain deeper knowledge on the ecology of these parasites and their bird host. A total of 2383 Contracaecum were collected from the gastric mucosa of 28 great cormorants (18 from Israel and 10 from Italy). A subsample was processed for morphological analyses in light and scanning electron microscopy (SEM), and for molecular analyses through amplification and sequencing of the ITS rDNA and the cox2 mtDNA, and through PCR-RFLP. All the 683 Contracaecum subjected to molecular identification belonged to C. rudolphii s.l., (300 C. rudolphii A and 383 C. rudolphii B). SEM micrographs provided, for the first time, details of taxonomic structures in male specimens from both sibling species, and the first SEM characterization of C. rudolphii B. This work presents the first data on the occurrence of sibling species of C. rudolphii in Israel and provides additional information on the distribution of C. rudolphii A and B in Italy, confirming the high prevalence and intensity of infection observed in Ph. carbo sinensis from other Italian areas.

Testicular Dirofilaria repens infection was identified and confirmed by sequence analysis in a child in northeastern Italy. Because human dirofilariasis is emerging in southern and eastern Europe, this parasitic infection should be considered in the differential diagnosis of scrotal swelling in disease-endemic countries to avoid unnecessary interventions, such as orchiectomy.

Electric fishing is an illegal hunting method, unfortunately widely used by poachers to paralyze fish and to catch many animals in a short time. In Italy, it is authorized only for scientific and conservative purposes. Between 2014 and 2018, the Ferrara section of the Experimental Zooprophylactic Institute of Lombardy and Emilia Romagna, Italy, received nine cases of potentially illegal electric fishing in Po river and its tributary rivers. Necropsies were performed following standard protocols and samples of different tissues were collected and examined using histochemical and immunohistochemical techniques. Gross lesions frequently observed were circulatory alteration phenomena (i.e. multi-organ hyperemia, hemorrhages and congestion, hemopericardium), also found histologically, in addition to multifocal degenerative and necrotic muscular processes that could be attributed to injuries from electric current, as already reported in literature. Immunohistochemical investigations confirmed degenerative and necrotic lesions with myoglobin depletion and a corresponding fibrinogen accumulation. Myoglobin globules were also detected in the renal parenchyma, as consequent of rhabdomyolysis. The results of this study allowed to correlate electric fishing to gross, histologic and immunohistochemical lesions, which together constitute a pathognomonic picture to be considered a reference standard in this type of illegal controversy.

Nervous Necrosis Virus (NNV) represents one of the most threatening pathogens for Mediterranean aquaculture. Several NNV strains are currently co-circulating in the Mediterranean Basin with a high prevalence of the RGNNV genotype and the RGNNV/SJNNV reassortant strain and a more limited diffusion of the SJNNV genotype and the SJNNV/RGNNV reassortant. In the present study, a one-step multiplex RT-PCR (mRT-PCR) assay was developed as an easy, cost-effective and rapid diagnostic technique to detect RGNNV and the reassortant RGNNV/SJNNV strain and to distinguish them from SJNNV and the reassortant SJNNV/RGNNV strain in a single RT-PCR reaction. A unique amplification profile was obtained for each genotype/reassortant enabling their rapid identification from cell culture lysates or directly from brain tissues of suspected fish. The method’s detection limit varied between 10 2.3 and 10 3.4 TCID 50 ml -1 depending on viral strains. No cross-reacitivty with viruses and bacteria frequently associated with gilthead seabream, European seabass and marine environment was observed. The mRT-PCR was shown to be an accurate, rapid and affordable method to support traditional diagnostic techniques in the diagnosis of VNN, being able to reduce considerably the time to identify the viral genotype or the involvement of reassortant strains.

The separation of Clinostomum complanatum Rudolphi, 1814 and Clinostomum marginatum Rudolphi, 1819 has long been unclear. Recent data confirm the validity of the junior species, C. marginatum, by ∼1% differences in its 18S rDNA sequences. We collected adults and metacercariae of C. complanatum and C. marginatum and found reliable morphological differences in the genital complex at both developmental stages. In addition, we identified basic morphometrics (distance between suckers, body width) in metacercariae that may be useful for discriminating the species. The morphological differences were supported by the comparison of sequences of internal transcribed spacers of ribosomal DNA and of the mitochondrial gene cytochrome c oxidase I (COI) from 39 specimens. In 36 specimens, the average divergence between the species was 7.3% in ITS and 19.4% in COI sequences. Two specimens from North America and 1 from Europe had sequences that did not allow them to be clearly allied with either species.

Three new species of Neoechinorhynchus are described from freshwater fish of Tigris River, at Salah Al-Din province, mid Iraq, namely Neoechinorhynchus tigrisensisn. sp. from Shilik Leuciscus vorax (Heckel) and Brond-snout Chondrostoma regium (Heckel), N. planilizain. sp. from Abu mullet Planiliza abu (Heckel) and Tigris catfish Silurus triostegus (Heckel) and N. barbin. sp. from Mesopotamian barb Capoeta damascina (Valenciennes). The first two new species shared the para-receptacle structure (PRS) with seven nominal valid species of the genus Neoechinorhynchus but they differed in other taxonomic traits, while the third new species shared the vaginal vestibular muscles (para vaginal patch) at the end of female trunk with N. spiramuscularis Amin, Heckmann & Ha, 2014 and N. zabensis Amin, Abdullah & Mhaisen, 2003 but differed for further morphological features. Proboscis hooks of middle and posterior circles in N. barbin. sp. are remarkably bigger than in the other two new species (N. tigrisensis and N. planilizai) while the hooks in first circle is almost similar in the three species. Furthermore, N. tigrisensisn. sp. and N. planilizain. sp. differ from some marine species by having smallest hooks size at anterior circle, with different shape and size of hooks. The three new species reported in the present study are compared with other eight species of Neoechinorhynchus reported so far from fish in Iraq. This work contributes to increase the knowledge on the biodiversity of fish parasites all over the world, especially for acanthocephalans.

The fluke Clinostomum complanatum , a parasite of piscivorous birds, but also reptiles and rarely mammals, has established several foci in the western Palaearctic regions. Previous studies pointed out the complicated taxonomy of the genus, but broader population genetic analysis of C. complanatum has not yet been carried out. The aim of this study was to determine the structure, intraspecific variability, and diversity of mitochondrial cox 1 haplotypes of C. complanatum from different localities in Slovakia (Danube floodplain forests) and Italy (Emilia-Romagna and Tuscany), as well as to evaluate the interrelationships among populations from Europe, the Middle East, and North Africa. The genetic structure of C. complanatum from Slovakia and Italy was represented by a great number of haplotypes, showing stable populations with high intraspecific diversity. The haplotypes of samples from other localities (Romania, Turkey, Egypt, and Iran) showed possible gene flow among the populations from Central Europe down to the Mediterranean region, North Africa, and the Middle East. The genetic homogeneity of these samples can be linked to the distribution and migratory routes of the definitive hosts, aquatic piscivorous birds, mainly herons and cormorants, that spread parasite eggs among the continents. La douve Clinostomum complanatum , un parasite des oiseaux piscivores, mais aussi des reptiles et rarement des mammifères, a établi plusieurs foyers dans les régions paléarctiques occidentales. Des études antérieures ont souligné la taxonomie compliquée du genre, mais une analyse génétique de la population plus large de C. complanatum n’avait pas encore été réalisée. L’objectif de cette étude était de déterminer la structure, la variabilité intraspécifique et la diversité des haplotypes mitochondriaux cox1 de C. complanatum de différentes localités de Slovaquie (forêts de la plaine inondable du Danube) et d’Italie (régions d’Émilie-Romagne et Toscane), ainsi que d’évaluer les interrelations entre les populations d’Europe, du Moyen-Orient et d’Afrique du Nord. La structure génétique de C. complanatum de Slovaquie et d’Italie était représentée par un grand nombre d’haplotypes, montrant des populations stables avec une grande diversité intraspécifique. Les haplotypes d’échantillons provenant d’autres localités (Roumanie, Turquie, Égypte et Iran) ont montré un possible flux génétique entre les populations de l’Europe centrale jusqu’à la région méditerranéenne, l’Afrique du Nord et le Moyen-Orient. L’homogénéité génétique de ces échantillons peut être liée à la distribution et aux voies migratoires des hôtes définitifs, des oiseaux aquatiques piscivores, principalement hérons et cormorans, qui propagent les œufs des parasites entre les continents.