Fatal infection with Taenia martis metacestodes in a ring-tailed lemur (Lemur catta) living in an Italian zoological garden

A case of fatal infection caused by larval forms of Taenia martis in a ring-tailed lemur (Lemur catta) living in the Rome zoological garden is described. The animal, living in a semi-natural pen with other 15 conspecific individuals and being fed with fresh fruit and vegetables, yoghurt and eggs, was transported to the Istituto Zooprofilattico of Rome for post-mortem examination. The anamnesis included, ten days before the death, apathy, lack of appetite, abdominal distension and diarrhoea. A severe exudative fibrinous-purulent peritonitis with numerous adhesions between the abdominal wall and the bowel loops was detected. After intestine removal, two free and viable, 4cm long, whitish, leaf-like parasitic forms were pinpointed. Macroscopic examination of the two parasites allowed their identification as larval stages of cestodes, identified via molecular analysis as T. martis metacestodes. This report represents the first record of T. martis infection in the host species and in a zoological garden and for the pathological relevance of the infection

Whole mitochondrial genome sequencing provides new insights into the phylogeography of loggerhead turtles (Caretta caretta) in the Mediterranean Sea

Population structure and phylogeography of the loggerhead sea turtle (Caretta caretta) have so far been assessed mainly by mitochondrial DNA (mtDNA) single-gene sequencing studies. However, phylogenetic relationships amongst matrilines, genetic characterisation of rookeries and mixed-stock analyses have suffered from the limited resolution obtained by comparison of relatively short sequences such as from the mtDNA control region. Whole mitogenome sequencing can significantly improve population genetics, particularly in marine organisms showing female natal philopatry. Despite mitogenomics becoming increasingly common in biodiversity monitoring and conservation, only a few complete mitogenomes are available for C. caretta. In this study, we sequenced the complete mtDNA of 61 loggerhead turtles sampled between 2008 and 2021 along the Italian coastline and central Mediterranean Sea. We assigned complete mtDNA haplotypes to dead embryos and bycatch samples, and introduced a first nomenclature for loggerhead mitogenomes. Analysis of mtDNA diversity, Maximum Parsimony and Bayesian phylogenetic reconstruction allowed improved resolution of lineages with respect to studies reporting on partial mtDNA control region sequence comparisons, and we were able to further inform previous analyses on loggerhead ancestry based on control region haplogroups. Overall, whole mitogenome analysis has potential for considerable improvement of evolutionary history and phylogeographic investigations as well as mixed-stock surveys of loggerhead turtles

Multidisciplinary studies on a sick-leader syndrome-associated mass stranding of sperm whales (Physeter macrocephalus) along the Adriatic coast of Italy

Mass strandings of sperm whales (Physeter macrocephalus) are rare in the Mediterranean Sea. Nevertheless, in 2014 a pod of 7 specimens stranded alive along the Italian coast of the Central Adriatic Sea: 3 individuals died on the beach after a few hours due to internal damages induced by prolonged recumbency; the remaining 4 whales were refloated after great efforts. All the dead animals were genetically related females; one was pregnant. All the animals were infected by dolphin morbillivirus (DMV) and the pregnant whale was also affected by a severe nephropathy due to a large kidney stone. Other analyses ruled out other possible relevant factors related to weather conditions or human activities. The results of multidisciplinary post-mortem analyses revealed that the 7 sperm whales entered the Adriatic Sea encountering adverse weather conditions and then kept heading northward following the pregnant but sick leader of the pod, thereby reaching the stranding site. DMV infection most likely played a crucial role in impairing the health condition and orientation abilities of the whales. They did not steer back towards deeper waters, but eventually stranded along the Central Adriatic Sea coastline, a real trap for sperm whales

Prion search and cellular prion protein expression in stranded dolphins.

The recent description of a prion disease (PD) case in a free-ranging bottlenose dolphin (Tursiops truncatus) prompted us to carry out an extensive search for the disease-associated isoform (PrPSc) of the cellular prion protein (PrPC) in the brain and in a range of lymphoid tissues from 23 striped dolphins (Stenella coeruleoalba), 5 bottlenose dolphins and 2 Risso s dolphins (Grampus griseus) found stranded between 2007 and 2012 along the Italian coastline. Three striped dolphins and one bottlenose dolphin showed microscopic lesions of encephalitis, with no evidence of spongiform brain lesions being detected in any of the 30 free-ranging cetaceans investigated herein. Nevertheless, we could still observe a prominent PrPC immunoreactivity in the brain as well as in lymphoid tissues from these dolphins. Although immunohistochemical and Western blot investigations yielded negative results for PrPSc deposition in all tissues from the dolphins under study, the reported occurrence of a spontaneous PD case in a wild dolphin is an intriguing issue and a matter of concern for both prion biology and intra/inter-species transmissibility, as well as for cetacean conservation medicine

A CASE OF PAPILLOMAVIRUS-ASSOCIATED TONGUE ADENOCARCINOMA IN A CAPTIVE WHITE RHINOCEROS

Papillomaviruses induce hyperplastic and tumoral lesions in different animal species4.We describe a case of Papillomavirus-Associated adenocarcinoma which occurred in a 41–year-old male white rhinoceros, maintained at the Zoological Garden of Pistoia, Tuscany3,5. The rhinoceros showed weight loss and difficulty in chewing the hay for three months before death2. At necroscopy, the most relevant finding was a lesion that involved the tongue, characterized by erosion of the mucosa with increased involvement of the dorsal surface. The cut surface showed lardaceous appearance with escape of purulent material. Histologically, the mass predominantly involved the submucosa and muscular portion of the tongue, resulting in closely packed large, medium, and small glands consisted of columnar cells without mucous cells. Tumor cells exhibited amphophilic to pale eosinophilic cytoplasm, high nucleo-cytoplasmic ratio and medium mitotic rate. No squamous differentiation was noted, and the squamous epithelium immediately adjacent to the adenocarcinoma showed slight perinuclear halos suggestive of PV-related changes. The tumor cells showed diffuse staining for A1-A3 pan-keratins, but staining for keratin 7 (CK7) clearly separated the adenocarcinoma from the adjacent non-neoplastic squamous epithelium. Keratin 19, keratin 20 and MUC-5AC were negative. Immunohistochemical staining performed with a monoclonal antibody against papillomaviruses evidenced a strong nuclear immunoreactivity only in glandular epithelium similarly to CK7. DNA extracted from paraffin-embedded tissue was tested by PCR using degenerated primers (FAP 59-64), amplifying common gene region of papillomaviral L11.The PCR result was an amplicon of 480 bp. Sequence similarity analysis with the BLAST tool of the National Center for Biotechnology Information and Papillomavirus Episteme tool showed that this fragments belongs to new putative PVs, not yet characterized for rhinoceros. To our knowledge this is the first case of Papillomavirus-Associated non-salivary gland-type adenocarcinoma arising in the base of the tongue in a captive white rhinoceros

Neuronal and astrocytic involvement in striped dolphins (Stenella coeruleoalba) with morbilliviral encephalitis.

Dolphin morbillivirus (DMV), a highly pathogenic agent, may cause peculiar, "brain-only" forms of infection (BOFDI), in which viral antigen and/or genome is found exclusively in the brain from striped dolphins (Stenella coeruleoalba). These BOFDIs show morphopathological similarities with subacute sclerosing panencephalitis and old dog encephalitis (ODE) in measles virus-infected patients and in canine distemper virus-infected dogs, respectively. The brain tissue from 3 BOFDI-affected striped dolphins was investigated by means of double labelling-indirect immunofluorescence (DL-IIF) and ultrastructurally, in order to characterize the DMV-targeted neuronal and non-neuronal cell populations, along with the associated submicroscopic findings. Viral colonization of calbindin-immunoreactive (IR) and nitric oxide synthase-IR neurons was detected in the cerebral parenchyma from the 3 DMV-infected dolphins under study, associated with nuclear (chromatin) and cytoplasmic (mitochondrial) ultrastructural changes. Furthermore, a limited viral targeting of brain astrocytes was found in these animals, all of which exhibited a prominent astrogliosis/astrocytosis. To the best of our knowledge, those herein reported should be the first submicroscopic pathology and neuropathogenetic data about BOFDI in striped dolphins. In this respect, the marked astrogliosis/astrocytosis and the low viral colonization of brain astrocytes in the 3 DMV-infected dolphins under investigation are of interest from the comparative pathology and viral neuropathogenesis standpoints, when compared with ODE-affected dogs, in whose brain a non-cytolytic, astrocyte-to-astrocyte infectious spread has been recently documented. Further studies aimed at characterizing the complex DMV-host interactions in BOFDI-affected striped dolphins are needed

Morbilliviral encephalitis in a striped dolphin Stenella coeruleoalba calf from Italy.

A free-living, striped dolphin Stenella coeruleoalba calf was stranded on the Latium coast of Italy in November 2009. Significant neuropathological findings included non-suppurative meningoencephalitis, microgliosis, neuronal degeneration, astrocytosis, and occasional multinucleate syncytia. Immunohistochemistry (IHC) and reverse transcription polymerase chain reaction (RT-PCR) for Morbillivirus were positive exclusively from the brain, with morbilliviral antigen and nucleic acid being detected in neurons and, to a lesser extent, in astrocytes. A low neutralizing antibody titer (1:10) against Morbillivirus (Canine distemper virus, CDV) was also found in blood serum, with no simultaneous presence of serum antibodies to Brucella spp. or Toxoplasma gondii. Furthermore, no pathogenic bacteria were isolated from any tissue or biological sample. This is the second report of morbilliviral encephalitis in a striped dolphin stranded along the Italian coastline in a 16 yr period (1993 to 2009). The neurohistopathological, IHC, and biomolecular features of this case are of additional interest, as antigenic and genomic positivity were exclusively confined to the brain of this dolphin, which may have acquired morbilliviral infection either postnatally or transplacentally

Prion search and cellular prion protein expression in stranded dolphins

The recent description of a prion disease (PD) case in a free-ranging bottlenose dolphin (Tursiops truncatus) prompted us to carry out an extensive search for the disease-associated isoform (PrPSc) of the cellular prion protein (PrPC) in the brain and in a range of lymphoid tissues from 23 striped dolphins (Stenella coeruleoalba), 5 bottlenose dolphins and 2 Risso s dolphins (Grampus griseus) found stranded between 2007 and 2012 along the Italian coastline. Three striped dolphins and one bottlenose dolphin showed microscopic lesions of encephalitis, with no evidence of spongiform brain lesions being detected in any of the 30 free-ranging cetaceans investigated herein. Nevertheless, we could still observe a prominent PrPC immunoreactivity in the brain as well as in lymphoid tissues from these dolphins. Although immunohistochemical and Western blot investigations yielded negative results for PrPSc deposition in all tissues from the dolphins under study, the reported occurrence of a spontaneous PD case in a wild dolphin is an intriguing issue and a matter of concern for both prion biology and intra/inter-species transmissibility, as well as for cetacean conservation medicine