Mass mortality event of round sardinella Sardinella aurita Valenciennes associated with Glugea Thélohan, 1891 microsporidian infection off the southern Italian coast.

Intracellular parasites of the genus Glugea Thélohan, 1891 (Microsporidia) comprise about 34 putative species capable of causing high morbidity and mortality in freshwater and marine teleost fishes. In this study, we report on the first mass mortality event associated with Glugea sp. infecting free-ranging round sardinella Sardinella aurita in the southern Tyrrhenian Sea (Italy). Here, we describe the ultrastructure of mature spores of this microsporidian and characterize it molecularly, as well as report its phylogenetic position. Most of the affected fish showed an irregular swelling of its abdomen. At necropsy, a variable number of xenomas, spherical to ellipsoidal in shape, were found in the peritoneal cavity strongly attached to the viscera of all fish. Histological analysis revealed varying severity of chronic inflammation along with occasional necrosis in visceral organs associated with multiple xenoma proliferation. These pathological findings were considered the main cause of this mass mortality event. Morphologically, the present material was closely related to G. sardinellesis and G. thunni. The phylogenetically closest taxa to the newly SSU rDNA sequence were G. thunni and an erroneusly identified  G. plecoglossi, which were very closely related to each other, also suggesting that all these sequences might belong to the same species.

Lightning deaths in three outdoor dogs: A case study.

This case report presents findings in three German Shepherd placed outdoor, dead after a night of thunderstorm.

Bovine delta papillomavirus E5 oncoprotein negatively regulates the cGAS-STING signaling pathway in cattle in a spontaneous model of viral disease.

Persistent infection and tumorigenesis by papillomaviruses (PVs) require viral manipulation of various cellular processes, including those involved in innate immune responses. The cyclic guanosine monophosphate-adenosine monophosphate synthase-stimulator of interferon genes (cGAS-STING) pathway has emerged as an essential innate immune sensing system, that recognizes DNA and trigger potent antiviral effector responses. In this study, we found that bovine PV (BPV) E5 protein, the major oncoprotein of bovine delta PVs, interacts with STING but not with cGAS in a spontaneous BPV infection of neoplastic urothelial cells of cattle. Real-time RT-PCR revealed a significant reduction in both cGAS and STING transcripts in E5-expressing cells. Furthermore, western blot (WB) analysis failed to detect any variation in the expression of interferon-inducible protein 16 (IFI16), an upstream effector of the STING pathway. A ternary complex composed of E5/STING/IFI16 was also observed. Co-immunoprecipitation studies showed that STING interacts with a protein network composed of total and phosphorylated TANK-binding kinase 1 (TBK1), total and phosphorylated interferon regulatory factor 3 (IRF3), IRF7, IKKα, IKKß, IKKϵ, ELKS, MEKK3, and TAK1. RT-qPCR revealed a significant reduction in TBK1 mRNA levels in BPV-infected cells. WB analysis revealed significantly reduced expression levels of pTBK1, which is essential for the activation and phosphorylation of IRF3, a prerequisite for the latter to enter the nucleus to activate type 1 IFN genes. WB also revealed significantly down-expression of IKKα, IKKß, IKKϵ, and overexpression of IRF7, ELKS, MEKK3, and TAK1in BPV-positive urothelial cells compared with that in uninfected healthy cells. Phosphorylated p65 (p-p65) was significantly reduced in both the nuclear and cytosolic compartments of BPV-infected cells compared with that in uninfected urothelial cells. Our results suggest that the innate immune signaling pathway mediated by cGAS-STING is impaired in cells infected with BPV. Therefore, effective immune responses are not elicited against these viruses, which facilitates persistent viral infection and subsequent tumorigenesis.

The Mediterranean Mussel (Mytilus galloprovincialis) as Intermediate Host for the Anisakid Sulcascaris sulcata (Nematoda), a Pathogen Parasite of the Mediterranean Loggerhead Turtle (Caretta caretta).

Sulcascaris sulcata (Anisakidae), a pathogenic nematode of sea turtles, may cause ulcerous gastritis with different degrees of severity. Previous studies demonstrated a high prevalence of infection in the Mediterranean loggerhead turtle (Caretta caretta), although no data on the potential intermediate hosts of this nematode has been published thus far from the Mediterranean basin. Here, using molecular analyses, we demonstrated that the cross sections of nematode larvae observed histologically in Mediterranean mussels (Mytilus galloprovincialis) collected from a farm along the Tyrrhenian coast of southern Italy belong to S. sulcata. The BLAST analysis of sequences at the ITS2 region of rDNA and mtDNA cox2 gene loci here obtained from samples of two Mediterranean mussels containing nematode larvae showed 100% homology with those at the same gene loci from the adults of S. sulcata collected from the Mediterranean Sea and deposited in GenBank. To our knowledge, this study is the first to present data on a potential intermediate host of S. sulcata in the Mediterranean basin and to report a nematode parasite from the Mediterranean mussel.

Correction to: Epidemiology of Sulcascaris sulcata (Nematoda: Anisakidae) ulcerous gastritis in the Mediterranean loggerhead sea turtle (Caretta caretta).

The original version of this article contained a mistake in the value of Bar in figure 3 caption. It should be Bar = 200 µm instead of Bar = 500 µm.

Epidemiology of Sulcascaris sulcata (Nematoda: Anisakidae) ulcerous gastritis in the Mediterranean loggerhead sea turtle (Caretta caretta).

Sulcascaris sulcata Rudolphi 1819 is a gastric nematode parasite of sea turtles. Here, we report the occurrence and describe for the first time the pathological changes caused by S. sulcata in the Mediterranean loggerhead sea turtle (Caretta caretta) stranded along the Tyrrhenian coast and northern Adriatic coast of Italy. Prevalence of infection was significantly higher in loggerhead sea turtles from the Adriatic Sea. Both prevalence and abundance of infection showed an increasing trend along with host age classes from both geographical localities. Nevertheless, while many small loggerhead sea turtles were found infected from the Adriatic Sea, only bigger individuals were infected from the Tyrrhenian Sea. The most common gross pathological change was a mucous gastritis with focal to multifocal raised ulcerous lesions roundish to irregular in shape ranging from 1 to over 20 cm in length, and cream-yellowish to greenish in color. The severity grade of gastritis increased with higher number of S. sulcata individuals. Microscopic pathological changes ranged from atrophic gastritis with heterophilic infiltration in the lamina propria to the destruction of the mucosal and sub-mucosal surfaces and necrosis. Results here obtained demonstrate that S. sulcata may cause ulcerous gastritis in both samples of loggerhead sea turtles studied from the Mediterranean Sea. Observed differences in S. sulcata infection among the different host age classes and between the two studied basins are likely linked to the differences of regional habitat and intermediate prey host availability.

Grillotia (Cestoda: Trypanorhyncha) plerocerci in an anglerfish (Lophius piscatorius) from the Tyrrhenian Sea.

Trypanorhynch cestodes are common parasites of marine fish with complicated life cycles which have been suggested as model taxa to study the evolution of marine helminth parasites and their life cycles. Among the Trypanorhyncha, the genus Grillotia includes 18 valid species, of which only four have been found in Mediterranean fish hosts. Morphological, histopathological, and molecular data are presented on a massive Grillotia plerocercus infection in an anglerfish (Lophius piscatorius) from the Tyrrhenian Sea. BLAST analysis of the 28S rDNA sequences revealed 99% similarity between specimens here found and a G. (Bathygrillotia) rowei sequence available in GenBank with a total of six nucleotide site differences. A morphological study suggested that the Grillotia sp. here reported did not match important characters to those previously reported from the Mediterranean Sea. Taking in account these differences, we prefer to place these specimens within Grillotia sensu lato until more material is available for study including sequences from adult specimens of Grillotia spp. from the Mediterranean Sea.

Coxiella burnetii in Infertile Dairy Cattle With Chronic Endometritis.

Coxiella burnetii is an obligate intracellular pathogen and the cause of Q fever in many animal species and humans. Several studies have reported the association between C. burnetii and abortion, premature delivery, stillbirth, and weak offspring. However, no solid evidence indicates that C. burnetii causes endometritis, subfertility, and retained fetal membranes. For this study, histopathological and PCR evaluation were performed on 40 uterine biopsies from dairy cattle with poor fertility. Uterine swabs were concurrently tested with microbiology assays. The endometrial biopsies of 30 cows did not have any significant lesions, and no pathogens were identified by aerobic bacterial culture and PCR. Ten cows were PCR-positive for C. burnetii and negative for other pathogens by aerobic bacterial culture and PCR. These 10 cases revealed a mild to severe chronic endometritis admixed with perivascular and periglandular fibrosis. Immunohistochemical evaluation of C. burnetii PCR-positive biopsies identified, for the first time, the presence of intralesional and intracytoplasmic C. burnetii in macrophages in the endometrium of cattle.

NEUROPATHOLOGIC FINDINGS IN CETACEANS STRANDED IN ITALY (2002-14).

We summarized the neuropathologic findings in 60 cetaceans stranded along the Italian coastline from 2002 to 2014. The following neuropathologic changes were detected in 45% (27/60) of animals: nonsuppurative meningo-encephalitides (30%, 18/60), nonspecific lesions (12%, 7/60), suppurative encephalitis (2%, 1/60), and neoplasm (2%, 1/60). No histologic lesions were found in 47% (28/60) of the specimens. Five (8%, 5/60) samples were unsuitable for analysis. Analysis with PCR detected Brucella spp., morbillivirus, and Toxoplasma gondii infection in one, six, and seven individuals, respectively. Immunohistochemical analysis confirmed positivity for morbillivirus and for T. gondii infection in three cases each. No evidence of the scrapie-associated prion protein PrPSc was detected. Our findings underscore the importance of an adequate surveillance system for monitoring aquatic mammal pathologies and for protecting both animal and human health.

Expression and activation of platelet-derived growth factor ß receptor, mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) and extracellular signal-regulated kinase (ERK) in canine mammary tumours.

Canine mammary tumours are frequent neoplasms mostly affecting intact female dogs, for which no 100% efficient therapy is available. Platelet derived growth factor ß receptor (PDGFßR) is a tyrosine kinase receptor (TKR) with a potential role in human breast cancer and a series of canine tumours. In this study we demonstrated, for the first time, expression of PDGFßR and its downstream transduction molecules, mitogen-activated protein/extracellular signal-regulated kinase kinase (MEK) and extracellular signal-regulated kinase (ERK), as well as their activated forms in canine mammary tumours by both biochemical analysis and immunohistochemistry. PDGFßR was expressed and hyperphosphorylated in the majority of tumour samples and tumour derived cell lines. Additionally, both MEK and ERK were expressed and activated in cell lines as well as biopsies. TKR inhibitors (TKRi) are currently under investigation as possible therapy in human breast and several canine tumours, thus our in vivo and in vitro findings pave the way for future studies aimed at establishing a potential therapeutic employment of TKRi for the treatment of canine mammary cancer.

Molecular identification and pathology of Anisakis pegreffii (Nematoda: Anisakidae) infection in the Mediterranean loggerhead sea turtle (Caretta caretta).

Nematodes of genus Anisakis spp. parasitize a wide range of marine hosts with marine mammals (mainly cetaceans) serving as definitive hosts, while fish, squid and other invertebrates serve as paratenic or intermediate hosts. Sea turtles can act as accidental or paratenic hosts for Anisakis spp. larvae, harbouring third-stage larvae unable to complete their life cycle in an ectothermic vertebrate. Post-mortem examination of 96 loggerhead sea turtles (Caretta caretta) stranded along the Italian coast of the Mediterranean Sea showed infection by Anisakis larvae Type I from 4 of 6 locations that were identified as belonging to Anisakis pegreffii by sequence analyses of the mtDNA cox2. Thirteen turtles (11 males and 2 females) were infected with A. pegreffii. Larvae were detected through gross necroscopy from 7 turtles, while in other 6 positive loggerhead sea turtles A. pegreffii larvae were revealed by histopathology. Pathological changes associated with A. pegreffii larvae in the stomach and intestine included necrosis and granulomatous response compatible with larvae migration. The role of the loggerhead sea turtle as an accidental host in the life cycle of this nematode is also discussed. This study is the first description of pathological changes associated with A. pegreffii in a sea turtle.