Cucullanus carettae Baylis, 1923, in a loggerhead sea turtle (Caretta caretta) from the Adriatic sea: first detection and molecular characterization.

Cucullanus carettae Baylis, 1923 (Nematoda: Cucullanidae) is found worldwide in loggerhead turtles (Caretta caretta). Regarding the Mediterranean, C. carettae has been identified in the Tyrrhenian and the Ionian Sea and a unique description of a Cucullanus sp. specimen in loggerheads from the Adriatic Sea has been reported in the literature so far. In the framework of a bio-monitoring project of the Abruzzo and Molise coasts, a parasitological survey was performed on stranded and by-caught sea turtles, at the Istituto Zooprofilattico of Abruzzo and Molise "G. Caporale." During necropsy, the gastrointestinal system of 72 stranded loggerhead turtles was analyzed for the presence of endoparasites and fecal samples were collected for coprological examination. Adult C. carettae (n = 123) was found in the upper intestine of one loggerhead turtle, associated with chronic lymphoplasmocytic enteritis. Additionally, five stool samples (6.9%) were positive for Cucullanus sp. eggs. Molecular characterization of adult nematodes was carried out to study phylogenetic relationships among the Cucullanus species. To our knowledge, this is the first morphological and molecular identification of C. carettae in loggerhead turtles from the Adriatic Sea. Additional studies on the distribution of this parasite in the Mediterranean are encouraged.

Morphological and molecular characterization of adults and larvae of Crassicauda spp. (Nematoda: Spirurida) from Mediterranean fin whales Balaenoptera physalus (Linnaeus, 1758).

Crassicauda boopis is known to infect the kidneys and vascular system of mysticetes included Balaenoptera physalus and has been recently reported in Mediterranean waters. Identification at the species level relies on the observation of morphological features of the adult parasites, but field conditions during necropsy and the massive reaction of the host's immune system often prevent optimal conservation of the extremities. Moreover, larval stages of Crassicauda have never been described and no sequences are available in public databases to help such identification. Adult and larvae of Crassicauda were isolated from four specimens of B. physalus and studied with morphological and molecular techniques. Specimens of C. anthonyi, C. grampicola and Crassicauda sp. isolated from Ziphius cavirostris, Grampus griseus, Stenella coeruleoalba and Tursiops truncatus respectively were studied as well. Sequences of nuclear markers 18S and ITS-2 and of mitochondrial gene cox1 were obtained and phylogenetic relationships within the genus Crassicauda were analysed. Analysis of the ITS2 grouped the different species in accordance with morphological identification, as already evidenced in literature for other Spirurida. A higher intra-specific variability was observed for the cox1 gene, for which two species (C. grampicola and C. anthonyi) did not appear as monophyletic in the tree. Well-developed non-attached larval specimens in the intestinal lumen of a whale calf were molecularly identified as C. boopis, allowing new insights on the life cycle of this species. This work broadens the genetic database on cetaceans parasites, allowing species identification even in challenging field conditions or in poor conservation of the samples; moreover, the first morphological description of C. boopis larvae is provided.

Parasitological and pathological findings in fin whales Balaenoptera physalus stranded along Italian coastlines.

Mediterranean fin whales Balaenoptera physalus face many threats to their conservation, including both anthropogenic and natural issues. There are few records of the parasitic fauna of this species in this geographical area. To partially fill in this gap of knowledge, we investigated the presence and potential impact of parasitic diseases in Mediterranean fin whales. Seven animals stranded along Italian coastlines between 2006 and 2015 were submitted for necropsy and parasitological examination. The protozoan parasite Toxoplasma gondii was detected in 1 fin whale and, for the first time in mysticetes, it was successfully genotyped as a type II strain with 15 microsatellite markers. One crustacean (Pennella spp.) and 4 helminth taxa (Crassicauda boopis, Ogmogaster antarcticus, Tetrabothrius ruudi and Bolbosoma sp.) were detected and morphologically identified. Different degrees of ectoparasitism by adult P. balaenoptera were recorded. Immature stages of Pennella sp. were also detected in 2 animals and are described here for the first time in cetaceans. Infestation by C. boopis was confirmed or suspected in 5 cases. Parasitic thrombi, involving renal veins and caudal vena cava, and fibrosis of renal parenchyma were associated with C. boopis and likely resulted in some degree of renal dysfunction. Larval nematodes were found within foci of mesenteric endarteritis. Further research to evaluate the prevalence of this potentially fatal endoparasitosis in Mediterranean fin whales is warranted.

Comparative field study to evaluate the performance of three different traps for collecting sand flies in northeastern Italy.

Three standard methods for collecting sand flies (sticky trap, CDC light trap, and CO2 trap) were compared in a field study conducted from June to October, 2012, at a site located in the center of a newly established autochthonous focus of canine leishmaniasis in northeastern Italy. Six traps (two sticky traps, two CDC light traps, and two CO2 traps) were activated at the same time for a single night every two weeks during the season of sand fly activity. A total of 5,667 sand flies were collected and 2,213 identified, of which 82.1% were Phlebotomus perniciosus, 17.4% P. neglectus, 0.3% Sergentomya minuta, and 0.2% P. mascitti. The performances of all traps were influenced by their position inside the site, increasing with proximity to the animal shelters. CO2 traps were more attractive for females of P. perniciosus and P. neglectus. CDC light traps showed an intermediate efficiency and were more attractive for P. neglectus, compared to other two traps. Results suggest that in northern Italy the CO2 trap is a suitable sampling method for sand fly monitoring programs that include transmitted pathogen surveillance.

Comparison of diagnostic methods to detect piroplasms in asymptomatic cattle.

This study was carried out to compare different diagnostic techniques to reveal the presence of piroplasms in asymptomatic cattle kept at pasture. Nineteen blood samples were collected from animals of two different areas of Emilia Romagna Region of Italy and processed for microscopic observation, PCR, serological test (IFAT) for Babesia bovis and Babesia bigemina antibodies and in vitro cultivation. The cultures were performed on both bovine and ovine erythrocytes. Seventeen blood smears (89%) were positive for piroplasms, while PCR was positive on 18 samples (95%). DNA sequencing of 18S rRNA identified the piroplasms as Theileria spp. In vitro cultures were successful for 6 samples (32%) cultured on bovine blood and subsequent identified these as Babesia major by PCR. On IFAT analyses of 16 samples, 36.8% resulted positive for B. bovis and 31.6% positive for B. bigemina. These results show, in the same animals, the co-infection with Babesia spp. and Theileria spp.; the detection of B. major was possible only using the in vitro cultures.

New insights into the epidemiology of bovine piroplasmoses in Italy.

Few studies have been published on bovine piroplasmoses in Italy, and therefore a clear picture of the epidemiology of these infections is difficult to obtain. Vertebrate and invertebrate hosts in Central and Northern Regions of Italy were investigated in 2005 and 2006, when microscopy, molecular tools and serological tests were applied to 468 blood samples drawn from cattle in order to evaluate the presence of these protozoa and identify possible risk factors. Ticks were also collected, identified and analyzed by molecular techniques. Microscopy identified 6.5% of the animals as positive, whereas PCR detected piroplasm DNA in 21.6%. BLAST analysis showed 67 amplicons (17.0%) referable to the Theileria sergenti/buffeli/orientalis group, 17 (4.3%) to Theileria annae, and 1 to Babesia divergens. Serology evidenced a prevalence of 45.4% for Babesia bovis, 17.4% for Babesia bigemina, and 34.9% for B. divergens. The 127 collected ticks were identified as belonging to 5 species, mostly represented by Rhipicephalus bursa, Hyalomma marginatum and Ixodes ricinus. Molecular analyses evidenced the presence of B. bovis and B. bigemina, in 3 and 5 ticks, respectively. Our findings suggest that different species of piroplasms are circulating in bovine populations in Central and Northern Italy, and provide new insights into the complex epidemiology of bovine piroplasmoses in Italy.

Parasitic diseases of marine fish: epidemiological and sanitary considerations.

Over recent decades, parasitic diseases have been increasingly considered a sanitary and economic threat to Mediterranean aquaculture. In order to monitor the distribution of parasites in cultured marine fish from Italy and study their pathogenic effects on the host, a three-year survey based on parasitological and histopathological exams was carried out on 2141 subjects from eleven fish species and coming from different farming systems (extensive, intensive inland farms, inshore floating cages, offshore floating cages and submersible cages). A number of parasitic species was detected, mostly in European sea bass (Dicentrarchus labrax), gilthead sea bream (Sparus aurata), mullets (Chelon labrosus, Mugil cephalus, Liza ramada) and sharpsnout sea bream (Diplodus puntazzo), with distribution patterns and prevalence values varying in relation to the farming system, in-season period and size category. The epidemiology and pathological effects of the parasites found during the survey are discussed.

Spinal deformity in a sandtiger shark, Carcharias taurus Rafinesque: a clinical-pathological study.

The present study investigated spinal deformity in a captive sandtiger shark and describes the clinical and histopathological features of the lesion. Clinically, the shark presented a marked spinal curvature in the region between the pectoral girdle and cranial dorsal fin. Radiographic studies revealed scoliosis and kyphosis of the vertebral column with a modification of the basic structure of five vertebrae. Post-mortem examination confirmed the results of radiographic studies and histological examination showed remodelling and cartilaginous proliferation of the vertebral bodies together with necrosis, haemorrhage and signs of a local chronic inflammatory response extending to the surrounding muscle. Different aetiologies are proposed and discussed but nutritional imbalance (dietary vitamin deficiency) and/or traumatic injury are considered the most likely contributing factors.