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The Northern raccoon Procyon lotor is a native mesocarnivore of North America which has been introduced to many Eurasian countries, where invasive populations currently occur. Regarding Italy, the species is present in Lombardy and in the Foreste Casentinesi National Park (Tuscany and Emilia-Romagna). In this work, we conducted a review on food habits of the Northern raccoon both in the native and in the introduced ranges, and we firstly reported the evidence on the impact of this species on native crayfish in Central Italy. We retrieved 18 papers published between 1932 and 2020, 14 from the native range and four from the invasive range. We analysed the frequency of occurrence of nine food categories (mammals, birds, reptiles, amphibians, fish, insects, crustaceans, fruits, and others). After fruits, crustaceans (where available) were the main food items detected in raccoon scats (up to 60% of the total diet, in the frequency of occurrence). In September 2019, we detected a total of 37 dead adult freshwater white-clawed crayfish Austropotamobius pallipes complex in Central Italy, with evident signs of predation by the Northern raccoon. The local population of Northern raccoon is fast expanding its range, whereas the local population of A. pallipes complex is declining. This crayfish species is protected by national and international laws. Thus, a monitoring program involving all stakeholders and institutions dealing with in environmental management, aiming at removing all raccoons currently present in this area of Central Italy, should be carried out in the short term to limit the potential impacts exerted by the fast-spreading of this invasive mammal. [ABSTRACT FROM AUTHOR]

The white-clawed crayfish Austropotamobius pallipes complex populations are decreasing in the Foreste Casentinesi, Monte Falterona and Campigna National Park (Central Italy), due to several factors, including illegal poaching, predatory fishes, drought, and invasive alien species. Recently, the Northern raccoon Procyon lotor has been reported to be present in the area of the National Park and has started to predate on the white-clawed crayfish. The aim of the study was to update the distribution and population status of A. pallipes in the reserves, other sites of the National Park, and surrounding areas to assess the potential effects of the raccoon. Crayfish were sampled by hand or by traps in 14 sites; sampled individuals were sexed and measured. Signs of raccoon presence (e.g., footprints and predated crayfish) were also recorded. Our study confirms the impact of raccoon on native crayfish: indeed, where the invasive mammal is present (six sites), crayfish disappeared, or their populations have been dramatically reduced in number, with a size distribution skewed towards juveniles. In two sites, close to urban settlements, fresh footprints of P. lotor and predated specimens of A. pallipes were also observed. Populations of crayfish are still abundant or even increasing as compared with samplings conducted in the past where raccoon is absent (five sites). Urgent actions (e.g., control of raccoons, and monitoring and restocking of A. pallipes populations if feasible and where possible) should be taken into account to guarantee the survival of this protected species.

The microsporidiosis of the endangered white-clawed crayfish Austropotamobius pallipes complex has generally been attributed to only one species, Thelohania contejeani , the agent of porcelain disease. Species identification was mostly assessed by macroscopic examination or microscopic evaluation of muscle samples rather than by molecular or ultrastructural analyses. A survey conducted on A. pallipes complex populations in Northern Italy highlighted the presence of two different microsporidia causing similar muscular lesions, T. contejeani and an undescribed octosporoblastic species Vairimorpha austropotamobii sp. nov. Mature spores and earlier developmental stages of V. austropotamobii sp. nov. were found within striated muscle cells of the thorax, abdomen, and appendages of the crayfish. Only octosporoblastic sporogony within sporophorous vesicles (SPVs) was observed. Diplokaryotic sporonts separated into two uninucleate daughter cells, which gave rise to a rosette-shaped plasmodium, and eight uninucleate spores were produced within the persistent SPV. Ultrastructural features of stages in the octosporoblastic sequence were similar to those described for Vairimorpha necatrix , the type species. Mature spores were pyriform in shape and an average of 3.9 × 2.2 µm in size. The polar filament was coiled 11–14 times, lateral to the posterior vacuole. The small subunit ribosomal RNA gene (SSU rRNA) and the large subunit RNA polymerase II gene (RPB1) of V. austropotamobii sp. nov. were sequenced and compared with other microsporidia. The highest sequence identity of SSU rRNA (99%) and RPB1 (74%) genes was with the amphipod parasite Nosema granulosis and subsequently with V. cheracis , which infects the Australian yabby Cherax destructor. In our work we discuss about the reasons for placing this new species in the genus Vairimorpha . In addition, we provide for T. contejeani a RPB1 gene sequence, supplemental sequences of SSU rRNA gene and ultrastructural details of its sporogony in the host A. pallipes complex. [ABSTRACT FROM AUTHOR]

Cases of co-occurrence among freshwater decapods are few. In their European range, the crab Potamon fluviatile and the white-clawed crayfish Austropotamobius pallipes complex do not usually share the same stream section or pond even if they live in sympatry, suggesting a strong segregation of the two species. The coexistence was hypothesized during the Pleistocene era, but without evidence of recent contact between them. The co-occurrence of these two species (i.e., occupying a common area) is reported in a stream of the Central Italy, being the first case in Italy and in Europe. To assess this co-occurrence, in 2015 we investigated three transects (only crabs, only crayfish and both species), collecting biotic and abiotic data. The three transects were similar for canopy cover, substrate composition and density of potential shelters (but not for the abiotic parameters), and characterized by a good water quality and abundance of preys, being suitable for the two species. No agonistic behavioural patterns have been observed, leading to hypothesize that co-occurrence may be favoured by the slightly smaller size of crayfish and crabs in the shared area than the size of animals in the other transects or by the different microhabitats chosen by crayfish and crabs when sharing the same stream section. Other possible explanations are discussed here, but further studies are necessary to better understand the co-occurrence of these two native crustaceans. Moreover, urgent measures are demanded to guarantee the survival of these protected species hampered by several threats. [ABSTRACT FROM AUTHOR]

Any initiative aimed at the management of a threatened species needs a good knowledge of its environmental requirements. Aims of this study were to individuate suitable areas for the reintroduction of crayfish species belonging to the Austropotamobius pallipes complex and to evaluate the causes of the local extinction in Tuscany (Italy) of some populations. Between May 2003 and September 2004, we collected ecological data from 19 streams of 4 catchments, 9 watercourses where crayfish were present (WI) and 10 where they were present in the recent past and absent now (WO), and we compared them. Multivariate analyses were performed using chemico-physical and biotic parameters to examine the relationships between their values and the presence of crayfish. The results did not allow us to find significant differences between the two categories of streams, suggesting their suitability for crayfish reintroduction.

Information was gathered on the distribution of white-clawed crayfish Austropotamobius pallipes complex in the watercourses of the Alessandria province (NW Italy), on the biological and ecological preferences of the species, and on the features of the biotopes in which it is found. A total of 409 sites on 361 watercourses connected to the main sub-basins of the Po River were analysed, with data gathered during the field research phase, conducted for three consecutive summers from 2002 through 2004. Thirteen percent of the sites investigated were found to currently house crayfish populations. The persistence in time of superficial water and natural morphology of the watercourse were found to be the chief requirements for the presence of crayfish. Data on land use, human density and other factors which potentially limit the survival of crayfish populations were also analysed.

The protection of freshwater biodiversity has become a priority task for conservation practices, as freshwater ecosystems host high levels of cryptic diversity, while also record similarly high rates of extinction. The Italian white-clawed crayfish Austropotamobius italicus is an endemic freshwater crustacean, threatened by several anthropogenic impacts such as habitat fragmentation, pollution, invasion of exotics, and climate change. Previous phylogenetic studies conducted in Italy pointed out a complex phylogeographic framework for the species, with four different subspecies currently recognized. Conservation efforts, particularly when involving restocking and reintroduction, require a detailed knowledge of their population genetics. In this study we describe the genetic structure of A. italicus populations in northern Italy (Lombardy Alpine foothills and northern Apennines) by using the informative mitochondrial marker cytochrome c oxidase subunit I, in order to assess their current evolutionary diversity and past phylogeographic history from a conservation perspective. Our results contribute to the mapping of the contact area among A. i. carsicus and A. i. carinthiacus in the Orobie Larian Prealps. More interestingly, we highlight the existence of two deeply differentiated evolutionary lineages within A. i. carsicus, showing alternative phylogeographic patterns and past demographic trends. We propose to consider these two clades as distinct molecular operational taxonomic units for the conservation of this endangered crayfish. [ABSTRACT FROM AUTHOR]

This is a study of movements and artificial shelters use by a 3-year established population of Austropotamobius pallipes complex, in a lentic environment in the Parco Lombardo della Valle del Ticino (NW Italy). Home-checking and mark-recapture methods were used to investigate movements of 178 crayfish and shelter use by 389 crayfish in summer and in autumn. Most crayfish (97.2%) moved but the movement was highly variable. The greater part of the distances moved was less than 10 m day-1. Multi-hole bricks with one opening have been mostly selected, and the lower level was preferentially occupied; frequently a single crayfish per brick was found. The animal's size affected the majority of the analysed variables: mean daily distance, area fidelity, level of shelter; on the other hand no case was significant for sex. The results highlight that this species is fairly mobile and could colonize waterbodies with similar features even rather rapidly. Mobility does not appear to affect the endangered status of this species. [ABSTRACT FROM AUTHOR]

Crayfish are regarded as useful indicators of environmental quality and freshwater biodiversity. However, reliable methods for monitoring their populations are needed so that this potential can be fully utilised. We report and discuss methodological aspects of the white-clawed crayfish (Austropotamobius pallipes complex) survey conducted in Piedmont, Italy, with the use of mark-recapture. The results suggest that the method can serve as a convenient tool for estimating the size of crayfish populations and inferring their temporal trends. The two populations investigated appeared closed except for wintertime and July. Consequently, the Robust Design, which is regarded as the most reliable mark-recapture approach, can be easily applied. The minimum effective sampling plan for monitoring purposes should comprise one primary period per year, conducted in the summer¿autumn season, and consisting of three capture sessions. If gaining insight into the ecology of the investigated species is the prime objective and sufficient resources are available, the optimal plan should include two primary periods (in spring and the summer¿ autumn season) of five capture sessions each. Capture sessions need to be separated by roughly 2-week intervals in order to avoid the strong, but short-term, negative effect of capturing crayfish on their recapture chances. As the model without heterogeneity in capture probabilities ensures better estimate precision we recommend that data collected for both sexes are analysed separately. Taking into consideration higher male catchabilities and sex ratio being invariably 1:1, it also seems beneficial to estimate only male numbers and double them to achieve total population sizes. [ABSTRACT FROM AUTHOR]

From 2004 to 2006, a parasitological survey aimed at the detection of the microsporidian parasite Thelohania contejeani Henneguy was carried out on 177 wild white-clawed crayfish (Austropotamobius pallipes complex) captured in six streams and rivers of the province of Belluno in north-eastern Italy. Microscopical examination of the skeletal muscles, and histological analysis applying different histochemical stains to full transverse and sagittal sections of the cephalothorax and abdomen were carried out. Transmission electron microscopy (TEM) was also conducted on the parasites recovered during the survey. Out of 177 crayfish examined, Thelohania contejeani (Microsporidia, Thelohaniidae) was present in only one crayfish from the Vena d’oro creek. The parasite was detected in the skeletal muscles in several developmental stages, including mature spores, which represented the most common stage recovered. Sporophorous vesicles were also present. Histological examination revealed that the fibres of the skeletal, cardiac and intestinal muscles were filled with spores. Melanin infiltrations were focally present in the infected striated muscles. The gill phagocytic nephrocytes were engulfed by small masses of spores. Among the staining techniques applied, Crossman’s trichrome stain represented the most effective method of detecting T. contejeani.

The Austropotamobius pallipes complex is formed by the two species A. pallipes and A. italicus, recently defined only on genetic basis, without obvious morphological features for their differentiation. Dealing with this problem, we have conducted a breeding experiment between these two presumed species under controlled conditions, in order to investigate the possible existence of reproductive barriers. Using 80 breeders (56 females and 24 males) collected from populations where their species identity was genetically well established, we performed four series of trials: Fita × Mpal and Fpal × Mita interspecific mating; Fita × Mita and Fpal × Mpal intraspecific mating as a control. We have monitored only the main phases of the reproduction in order to avoid excessive interference for the crayfish. In November 2009 the 91.7% of females having intraspecific mating, as well as the 100% of females having interspecific mating, produced eggs. In June 2010 the eggs from the 36.4% of females having intraspecific mating, and those from the 42.3% of females having interspecific mating did hatch. We emphasize the hatching of hybrids and their vitality.