Acorn-related acquired pseudomelanosis in Calabrian black pigs.

BACKGROUND: Melanosis of lymph nodes in black pigs has generally been related to regression of congenital melanoma and, occasionally, to ingestion of acorns. The aim of this manuscript is to confirm the hypothesis of a possible acquired acorn-related pseudomelanosis in the Nero Calabrese pig, a swine breed belonging to the group of Italian native breeds and whose coverage area corresponds to the region of Calabria, southern Italy. This pig is characterized by slow-growing subjects, producing, however, high quality meat suitable for the production of sausages and fine hams. The study was carried out on 142 normally slaughtered pigs. All organs were examined. Lymph nodes and intestine (jejunum) were sampled. Histochemistry was performed on deparaffinized histological sections to identify the cell types involved and to characterize the pigment stored. To further confirm the pigmentation disorder, immunohistochemistry was carried out. Total phenolic substances were identified in acorns through the use of a biochemical reaction. RESULTS: Lymph node pigmentation appears directly related to acorn ingestion, with a higher incidence in the group which was 70% natural fed (acorn of Quercus virgiliana). Moreover, findings obtained revealed how different amounts of phenolic substrates present in Q. virgiliana and Q. ilex acorns can influence the incidence of such exogenous pigmentation. CONCLUSION: The findings obtained in this study confirm the acquired nature of the melanin-like pigmentation detected in lymph nodes from acorn-fed swine. Acquired pigmentation must be differentiated from true melanosis as well as from melanosis related to tumor regression of congenital melanoma. This thesaurismosis can be proposed as a marker of wellbeing and quality, confirming that the pigs have been bred and fed in natural conditions.

In Vitro Simulated Hemoperfusion on Seraph®-100 as a Promising Strategy to Counteract Sepsis.

Blood purification represents a treatment option for sepsis, improving inflammation and the hyper-activated immune system. This study investigates the binding efficacy of Seraph®-100 against 108 CFU/mL of Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa), and Escherichia coli (E. coli) during a simulated hemoperfusion treatment. The fluorescence-activated cell sorting (FACS) technique was used to evaluate the bacteria reduction, whereas kinetic analysis and cultures revealed bacterial detection and counting at established time points. At the end of the experiment, the filter was cut at three different levels, obtaining suspensions for cultures and scanning electron microscopy (SEM) analyses. The FACS technique revealed a 78.77% reduction of the total bacterial load at the end of the treatment, with maximum filter sequestration occurring in the first 30 min of the treatment. Non-linear regression analysis of kinetic experiments (T0-240 min) highlighted a lower growth rate of S. aureus than the other two Gram bacteria, demonstrating a greater affinity without influencing a reduction rate of 99% for all three bacteria. The analyses of the suspension aliquots of the filter sections confirmed these data, revealing 1 × 108 CFU/mL, equal to the initial bacterial charge. Furthermore, the filter head adsorbed approximately 50% of bacteria, whereas the remaining amount was equally distributed between the body and the tail, as corroborated by SEM analysis. In conclusion, Seraph®-100 adsorbed 108 CFU/mL of S. aureus, E. coli, and P. aeruginosa during an in vitro simulated hemoperfusion session.

Cutaneous Malignant Melanoma with Testicular Metastases in a Wild Rabbit (Oryctolagus cuniculus).

Melanocytic skin tumours have been rarely described in pet rabbits, and exposure to UV light in sparsely haired areas has been hypothesised to play a cancerogenic role. Here, we describe a case of cutaneous malignant melanoma arising from the skin of the scrotum in an 8-year-old male wild rabbit, with testicular metastases as an unusual metastatic site for melanoma reported in humans to date. The tumour was nearly 5 cm in size, firm, and highly pigmented, with multifocal superficial ulcerations and large areas of intratumoural necrosis. The adjacent testis was 1.5 cm, multinodular, and black, obscuring tissue morphology. Histologically, the dermis was expanded by an infiltrative, densely cellular neoplasm composed of nests and sheets of polygonal to spindle neoplastic melanocytes, supported by scant fibrovascular stroma. Neoplastic cells showed intermediate N/C ratio, moderate basophilic cytoplasm, often obscured by abundant brownish granular pigment, and eccentric nuclei with prominent nucleoli. Cellular pleomorphism and nuclear atypia were severe, and high mitotic activity was observed. Diffuse dermal lymphovascular invasion was also observed. The testis was delimited by a thin tunica albuginea, and the parenchyma was largely obscured in its morphology by densely packed neoplastic cells. Seminiferous tubules, lined with a thin basement membrane and containing neoplastic and scattered spermatogenic cells, were occasionally observed. Neoplastic cells within the skin and the testis were positive for HMB-45, Melan-A, and S-100. The growing popularity of rabbits as pets allows for a greater ability to accumulate data on the spontaneous occurrence of tumours in these animals. Furthermore, descriptions of the biological aspects of spontaneously occurring tumours may serve to improve current knowledge in animal species and humans in which the same neoplasm may occur.

Upregulation of miR-21 and pro-inflammatory cytokine genes IL-6 and TNF-α in promoting a pro-tumorigenic microenvironment in canine mammary carcinomas.

This study evaluated the gene expression of the pro-inflammatory cytokines, interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in canine mammary tumors (CMTs), and correlated them with gene expression of miRNAs expected to regulate the secretion of pro-inflammatory cytokines within the tumor microenvironment (TME). Furthermore, gene expression of cytokines and miRNAs involved in tumor cell proliferation and invasion (i.e. miR-21; miR-124; miR-145) were correlated with tumor proliferation index (Ki67 index) to determine the prognostic value in CMTs. Twenty-six canine mammary samples were used, including 22 CMTs and 4 control samples. MiR-21, IL-6 and TNF-α were upregulated in mammary carcinomas compared with controls (p < 0.05). MiR-146b was downregulated in CMTs compared with control cases (p < 0.05). IL-6 expression showed a significant positive correlation with miR-21 and a negative correlation with miR-146b; while, TNF-α gene expression was positively correlated with miR-21 and miR-145 in mammary carcinomas. In carcinomas, the Ki67 index correlated positively with gene expression of IL-6 and miR-21 and negatively correlated with miR-145 and miR-146b. Specifically, gene expression of IL-6 and miR-21 was positively correlated with ki67 index >33.3%, whereas, expression of miR-145 and miR-146b was negatively correlated with ki67 index <33.3%. Results reinforce the concept of interaction between tumor cells and inflammatory cells within the TME, with a central role of IL-6 and TNF-α. Since the upregulation of miR-21 reflects the gene overexpression of interleukins and the high proliferation index of tumor cells, this miRNA may be considered a biomarker with prognostic value in CMTs.

Epigenetics in Canine Mammary Tumors: Upregulation of miR-18a and miR-18b Oncogenes Is Associated with Decreased ERS1 Target mRNA Expression and ERα Immunoexpression in Highly Proliferating Carcinomas.

The expression of miRNAs is one of the main epigenetic mechanisms responsible for the regulation of gene expression in mammals, and in cancer, miRNAs participate by regulating the expression of protein-coding cancer-associated genes. In canine mammary tumors (CMTs), the ESR1 gene encodes for ERα, and represents a major target gene for miR-18a and miR-18b, previously found to be overexpressed in mammary carcinomas. A loss in ERα expression in CMTs is commonly associated with poor prognosis, and it is noteworthy that the downregulation of the ESR1 would appear to be more epigenetic than genetic in nature. In this study, the expression of ESR1 mRNA in formalin-fixed, paraffin-embedded (FFPE) canine mammary tumors (CMTs) was evaluated and compared with the expression levels of miR18a and miR18b, both assessed via RT-qPCR. Furthermore, the possible correlation between the miRNA expression data and the immunohistochemical prognostic factors (ERα immunoexpression; Ki67 proliferative index) was explored. A total of twenty-six FFPE mammary samples were used, including 22 CMTs (7 benign; 15 malignant) and four control samples (three normal mammary glands and one case of lobular hyperplasia). The obtained results demonstrate that miR-18a and miR-18b are upregulated in malignant CMTs, negatively correlating with the expression of target ESR1 mRNA. Of note, the upregulation of miRNAs strictly reflects the progressive loss of ERα immunoexpression and increased tumor cell proliferation as measured using the Ki67 index. The results suggest a central role of miR-18a and miR-18b in the pathophysiology of canine mammary tumors as potential epigenetic mechanisms involved in ERα downregulation. Moreover, as miRNA expression reflects ERα protein status and a high proliferative index, miR-18a and miR-18b may represent promising biomarkers with prognostic value. More detailed investigations on a larger number of cases are needed to better understand the influence of these miRNAs in canine mammary tumors.

Intraspecific variability of the saccular and utricular otoliths of the hatchetfish Argyropelecus hemigymnus (Cocco, 1829) from the Strait of Messina (Central Mediterranean Sea).

Mesopelagic species are enjoining increasing attention due to the growing impact of fisheries activities on deep marine biocenosis. Improving the knowledge base on mesopelagic species is required to enhance their conservation due to the knowledge gaps regarding many species and families. In this context, otoliths can be fundamental to assessing their life history, ecomorphological adaptation to the deep environment and stock composition. The present paper aims to explore the saccular and utricular otoliths morphology and intra-specific variability of the hatchetfish, Argyropelecus hemigymnus, from the Strait of Messina. Lapilli and sagittae were collected from 70 specimens and separated into four size classes. Morphometric, shape and SEM investigations were performed to describe their morphology, contours, and external structural organization, also studying their intraspecific variability related to sample sizes and differences between otolith pairs. Results showed an otolith morphology different from those reported in the literature with fluctuating asymmetry in sagittae and lapilli belonging to Class IV, and a high otolith variability between all the size classes. Data herein described confirm the otoliths singularity of the population from the Strait of Messina, shaped by a unique marine environment for oceanographic and ecological features.

Systemic candidiasis in farm-reared red-legged partridges (Alectoris rufa) caused by Leucosporidium spp.

BACKGROUND: This report describes the results of radiological, histological and molecular examination of three farm-reared red-legged partridges (Alectoris rufa) affected by candidiasis. CASE PRESENTATION: Three juvenile farm-reared red-legged partridges in a batch of 100 of the same species were sent for clinical and pathological investigations. The owner referred of a sudden isolation of the sick animals, with apathy, diarrhea, ruffled plumage and respiratory rattles. Post mortem total body lateral projection radiograph showed an increased perihilar interstitial pattern and air bronchogram signs due to lung edema. At necropsy, carcasses showed cachexia; the pericloacal region was soiled by diarrheic fecal material. From the mouth to the intestine, a mucous yellowish fluid was present on a slightly reddish mucosa. Histopathology showed slight edema and congestion with different free fungal elements, referable to blastospores, hyphae and pseudohyphae. Biomolecular exam identified the most similar sequences as belonging to Leucosporidium scottii. CONCLUSION: To our knowledge, this case report describes for the first time this fungal species as a causative agent of candidiasis in birds.

Beta-catenin expression in pilomatrix carcinoma with multiple visceral metastases in a dog.

Beta-catenin is a protein initially identified as a submembrane component of the E-cadherin-mediated cell-to-cell adhesion system. It plays a role as a transcriptional factor in the wingless/Wnt signalling pathway. Beta-catenin has been associated with oncogenic activity in human benign and malignant pilomatrix neoplasms where the immunohistochemical profile of ß-catenin expression displayed both nuclear and cytoplasmic staining in basaloid cells. In this study, an 8-year-old female Irish setter dog was examined because of the presence of skin nodules. Tissue biopsies from different nodules were obtained and histological examination suggested a diagnosis of pilomatrix carcinoma. The dog spontaneously died after 2 months and necropsy showed multiple metastases in the nasal cavity, lungs, heart, kidney, liver and colon. Routine histopathology of metastatic sites showed features consistent with the pattern of primary neoplastic nodules. Immunohistochemical detection of ß-catenin was performed in both the primary tumour and the metastases. Beta-catenin expression was located in the nuclei, cytoplasm and membrane of squamoid cells and in the cytoplasm of basaloid cells, while shadow cells were completely negative. To the best of our knowledge, these data represent the first report on the immunohistochemical expression profile of ß-catenin in canine pilomatrix carcinoma.

RT-qPCR Expression Profiles of Selected Oncogenic and Oncosuppressor miRNAs in Formalin-Fixed, Paraffin-Embedded Canine Mammary Tumors.

MicroRNAs (miRNAs) can act as oncogenes or oncosuppressor genes, and their involvement in nearly all cancer-associated processes makes these small molecules promising diagnostic and prognostic biomarkers in cancer, as well as specific targets for cancer therapy. This study aimed to investigate the expression of 7 miRNAs (miR-18a, miR-18b, miR-22, miR-124, miR-145, miR-21, miR-146b) in formalin-fixed, paraffin-embedded canine mammary tumors (CMTs) by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Twenty-six mammary samples were selected, including 22 CMTs (7 benign; 15 malignant) and 4 control samples (3 normal mammary gland and 1 case of lobular hyperplasia). Oncogenic miR-18a, miR-18b and miR-21 were significantly upregulated in malignant tumors compared with control tissues (p < 0.05). Conversely, oncosuppressor miR-146b was significantly downregulated in benign and malignant mammary tumors compared with control samples (p < 0.05) while, no group-related differences in the expression levels of miR-22, miR-124 and miR-145 were found (p > 0.05). Upregulated miRNAs found here, may regulate genes involved in receptor-mediated carcinogenesis and proteoglycan remodeling in cancer; while miRNA with reduced expression can regulate genes involved in Toll-like receptor and MAPK signaling pathways. According to the results obtained in the current study, the oncogenic and oncosuppressor miRNAs analyzed here are dysregulated in CMTs and the dysregulation of miRNA targets may lead to specific altered cellular processes and key pathways involved in carcinogenesis. Of note, since oncogenic miRNAs predicted to regulate neoplastic cell proliferation and hormonal activities, they may play an active role in neoplastic transformation and/or progression, having mechanistic and prognostic relevance in CMTs.

Aspiration Pneumonia with Prominent Alveolar Mineralization in a Dairy Cow.

A 2-years-old Jersey breed cow showed severe respiratory distress and prolonged lateral recumbency about 1 week after parturition. The cow was regularly vaccinated against the main respiratory pathogens and was given a calcium salt solution per os shortly after calving in order to prevent milk fever. Despite therapy with broad-spectrum antibiotics and anti-inflammatory drugs, the cow spontaneously died one week later and was necropsied. On gross examination, a severe, acute, diffuse fibrinonecrotic tracheitis was observed. In addition, the cranioventral portions of both lungs appeared firm and severely congested, while the pleural surface was covered by a discrete amount of fibrinous exudate. Microscopically, the following lesions were observed: tracheal hemorrhages, acute, fibrinonecrotic and suppurative tracheitis, pulmonary hemorrhages, fibrinous bronchopneumonia and fibrinous pleuritis. Noteworthy, multiple foci of mineralization were observed, scattered throughout the lung parenchyma and occasionally within the tracheal mucosa. The presence of calcium deposits was confirmed by means of Von Kossa staining method. Based on clinical history, clinical signs and pathological findings, aspiration pneumonia caused by the accidental inhalation of liquid calcium salt supplement was diagnosed. The present case report highlights the relevance of the staff training to optimize animal production and welfare.

Postmortem Electrical Conductivity Changes of Dicentrarchus labrax Skeletal Muscle: Root Mean Square (RMS) Parameter in Estimating Time since Death.

Electric impedance spectroscopy techniques have been widely employed to study basic biological processes, and recently explored to estimate postmortem interval (PMI). However, the most-relevant parameter to approximate PMI has not been recognized so far. This study investigated electrical conductivity changes in muscle of 18 sea bass specimens, maintained at different room temperatures (15.0 °C; 20.0 °C; 25.0 °C), during a 24 h postmortem period using an oscilloscope coupled with a signal generator, as innovative technology. The root mean square (RMS) was selected among all measured parameters, and recorded every 15 min for 24 h after death. The RMS(t) time series for each animal were collected and statistically analyzed using MATLAB®. A similar trend in RMS values was observed in all animals over the 24 h study period. After a short period, during which the RMS signal decreased, an increasing trend of the signal was recorded for all fish until it reached a peak. Subsequently, the RMS value gradually decreased over time. A strong linear correlation was observed among the time series, confirming that the above time-behaviour holds for all animals. The time at which maximum value is reached strongly depended on the room temperature during the experiments, ranging from 6 h in fish kept at 25.0 °C to 14 h in animals kept at 15.0 °C. The use of the oscilloscope has proven to be a promising technology in the study of electrical muscle properties during the early postmortem interval, with the advantage of being a fast, non-destructive, and inexpensive method, although more studies will be needed to validate this technology before moving to real-time field investigations.

Environmental Toxicity Assessment of Sodium Fluoride and Platinum-Derived Drugs Co-Exposure on Aquatic Organisms.

Pharmaceuticals are widely acknowledged to be a threat to aquatic life. Over the last two decades, the steady use of biologically active chemicals for human health has been mirrored by a rise in the leaking of these chemicals into natural environments. The aim of this work was to detect the toxicity of sodium fluoride (NaF) exposure and platinum-derived drugs in an ecological setting on aquatic organism development. From 24 to 96 h post-fertilization, zebrafish embryos were treated to dosages of NaF 10 mg/L-1 + cisplatin (CDDP) 100 µM, one with NaF 10 mg/L-1 + carboplatin (CARP) 25 µM, one with NaF 10 mg/L-1 + CDDP 100 µM + CARP 25 µM. Fluoride exposure in combination with Cisplatin and Carboplatin (non-toxic concentration) had an effect on survival and hatching rate according to this study. Additionally, it significantly disturbed the antioxidant defense system and increased ROS in zebrafish larvae. NaF 10 mg/L-1 associated with CDDP 100 µM and CARP 25 µM, increased the production of apoptosis-related proteins (caspase 3, bax, and bcl-2) and the downregulation of acetylcholinesterase (AChE) activity, while no effect was seen for the single exposure.

Experimentally induced melanin-like pigmentation (thesaurismosis) related to acorn ingestion in Nero Siciliano pigs.

In this study, an experimental challenge was carried out by feeding Nero Siciliano pigs with acorn to evoke melanin-like pigmentation and support the hypothesis that it is caused by ingested material. Twelve pigs were studied, 6 fed with acorns and 6 fed commercial feed. At slaughter, all the animals fed on acorns showed black discolouration of almost all lymph nodes. The lymph nodes were normal in size and shape. Histochemical tests performed on tissues allowed us to identify and differentiate the pigment. Immunohistochemical staining for macrophage markers showed macrophages containing a variable amount of melanin-like granules. Our data, and the well-known steps of melanin formation, confirm that swine enzymes could act on phenolic substances found in acorns.

Penicilliosis in an African grey parrot (Psittacus erithacus).

Penicillium mycosis, or penicilliosis, is rare in animals. This report describes the clinical signs and pathological, immunohistochemical, and biomolecular findings in a case of avian penicilliosis in a Congo African grey parrot. On gross necropsy, pyogranulomatous changes were identified in the lung, liver, and kidney. Histopathology demonstrated periodic acid Schiff-(PAS) and Grocott-positive hyphae in these tissues. Immunohistochemistry confirmed the genus of the fungus and polymerase chain reaction (PCR) confirmed a high homology with Penicillium chrysogenum.

Mycobacterium tuberculosis in a red-crowned parakeet (Cyanoramphus novaezelandiae).

An adult female red-crowned parakeet (Cyanoramphus novaezelandieae) was presented for necropsy and histopathologic evaluation. The bird had died after exhibiting lameness, weight loss, respiratory signs, and hemoptysis. Postmortem radiographs revealed lesions in the diaphysis of the left femur and soft-tissue opacities in the lungs. At necropsy, the muscles of the left femur were pale and swollen, white-yellow small nodules were visible in lungs and air sacs, and the liver and other coelomic organs appeared pale. On histologic examination, areas of necrosis in the lungs were extensively infiltrated with acid-fast positive bacilli surrounded by macrophages, epithelioid cells, and giant cells. Acid-fast bacilli were also present in the left leg muscle and in granulomas in the liver, kidneys, and intestine. Fungal hyphae associated with a Splendore-Hoeppli phenomenon were visible in the left leg muscle. Mycobacterium tuberculosis was confirmed by polymerase chain reaction testing performed on muscle samples. Results were indicative of infection with M tuberculosis complicated by mycotic myositis. The disease in avian species is of zoonotic importance, and infected birds may be a useful sentinel for human infection.

A case of neoplastic synchronism in a dog.

INTRODUCTION: Synchronous primary tumors are considered severe, comorbid conditions in people representing neoplasm that develop independently and concomitantly. A diagnosis of synchronous tumors was made in a dog, underlying the difficulties to reach it without the aid of multiple diagnostic techniques aimed to demonstrate the simultaneous coexistence of different tumor types. MATERIALS AND METHODS: A 7-year-old male Boxer dog presented several tumors located on the skin of the left hind limb and the scrotal region. Moreover, additional tumors in the testicles, after palpation and ultrasound examination, were detected. Following diagnostic results, the cutaneous tumor, scrotum, and testes were surgically removed. RESULTS: Pathological investigations revealed the presence of five different tumors: a cutaneous mast cell tumor; a scrotal melanocytoma; three testicular neoplasms (Sertoli Sustentacular cell tumor, seminoma, and interstitial Leydig cell tumor). CONCLUSIONS: The present report describes a neoplastic synchronism due to the presence of five different primary tumors in a dog and, for the first time the presence of a collision testicular tumor together with other non-testicular primary tumors. The occasional finding underlines the importance of the knowledge of such conditions in the process of decision-making and in carrying out all the proper diagnostic procedures for a correct diagnosis and clinical staging.

Anti-Atherogenic Effect of 10% Supplementation of Anchovy (Engraulis encrasicolus) Waste Protein Hydrolysates in ApoE-Deficient Mice.

Fish protein consumption exerts beneficial metabolic effects on human health, also correlating with a decreased risk for cardiovascular disease. Fish waste contains high amount of proteins and utilization may offer the opportunity for generating compounds advantageous for human health. Especially, fish waste protein hydrolysates beneficially influence pathways involved in body composition, exerting anti-inflammatory and antioxidant activities, making their potential supplementation in human disorders of increased interest. This study assessed the effect of a 10% (w/w) anchovy waste protein hydrolysate (APH) diet for 12 weeks in reducing atherosclerosis in ApoE-/- mice, through histological and immunohistochemical methods. In addition, monitoring of plaque development was performed, using high-frequency ultrasound and magnetic resonance imaging. Overall, the APH diet attenuated atherosclerotic plaque development, producing a regression of arterial lesions over time (p < 0.05). Twelve weeks on an APH diet had an anti-obesity effect, improving lipid metabolism and reducing hepatic enzyme activity. A significant reduction in plaque size and lipid content was observed in the aortic sinus of APH-fed mice, compared to the control (p < 0.001), whereas no differences in the extracellular matrix and macrophage recruitment were observed. Supplementation of APH significantly attenuates atherosclerosis in ApoE-/- mice, exerting a lipid-lowering activity. The opportunity to use fish waste protein hydrolysates as a nutraceutical in atherosclerosis is worthy of future investigations, representing a low cost, sustainable, and nutritional strategy with minimal environmental impact.

Aspergillus Section Fumigati Pneumonia and Oxalate Nephrosis in a Foal.

Equine pulmonary aspergillosis is a rare deep mycosis often due to the hematogenous spread of hyphae after gastrointestinal tract disease. We describe herein the main clinic-pathological findings observed in a foal, which spontaneously died after showing diarrhea and respiratory distress. Necropsy and histopathological investigations allowed to diagnose pulmonary aspergillosis, which likely developed after necrotic typhlitis-colitis. Biomolecular studies identified Aspergillus section Fumigati strain as the causative agent. Notably, severe oxalate nephrosis was concurrently observed. Occasionally, oxalate nephropathy can be a sequela of pulmonary aspergillosis in humans. The present case report suggests that the renal precipitation of oxalates can occur also in horses affected by pulmonary aspergillosis and could likely contribute to the fatal outcome of the disease.

First Isolation and Molecular Characterization of Pseudorabies Virus in a Hunting Dog in Sicily (Southern Italy).

Pseudorabies virus (PrV) is the etiological agent of Aujeszky's disease, a viral infection that causes neurological lethal illness in mammals other than swine. Herein, we describe the occurrence of PrV infection in a hunting dog that had been bitten by an infected wild boar in Sicily, reporting for the first time genetic and phylogenetic data on the virus strain isolated in a dog in this Italian region. The dog was referred for severe neurological signs, respiratory distress, and intense itch around the muzzle. Death occurred within 48 h to the onset of clinical signs. On gross examination, self-induced skin lesions to the head due to intense itching and diffuse cerebral congestion were observed, whereas mild, aspecific, nonsuppurative meningitis was histologically diagnosed. Diffuse PrV positivity in neurons of the brainstem was observed by immunohistochemistry. PrV DNA was isolated and amplified from olfactory bulbs by nested PCR, targeting the viral glycoprotein G gene, and the sequence obtained matched with sequences of PrV isolates from dogs and wild boar. Isolation of PrV in the dog herein analysed denotes the spread of the virus in wild boar populations in Sicily and provides a proof of direct interspecies transmission. Thus, there is an urgent need to increase our understanding of the epidemiology of the PrV infection in wildlife to provide tools to trace possible spill over into domestic pigs or other livestock.

Intra- and interspecific variability among congeneric Pagellus otoliths.

Otolith features are useful tools for studying taxonomy, ecology, paleontology, and fish biology since they represent a permanent record of life history. Nevertheless, the functional morphology of otoliths remains an open research question that is useful to completely understand their eco-morphology. This study aims to deepen the knowledge of intra- and interspecific variation in sagitta morphology in three congeneric seabreams, to understand how such variability could be related to the lifestyles of each species. Therefore, the sagittae (n = 161) of 24 Pagellus bogaraveo, 24 Pagellus acarne, and 37 Pagellus erythrinus specimens, collected from the south Tyrrhenian Sea, were analyzed using scanning electron microscopy and a stereomicroscope to assess morphometric features, variability between otolith pairs and the external crystalline structure the of sulcus acusticus. Statistical analysis demonstrated that, between the species, variability in sagittal otolith rostral length growth and sulcus acusticus features, correlated with increased fish total length and body weight. Moreover, slight differences between otolith pairs were detected in P. acarne and P. erythrinus (P < 0.05). The results confirm changes in otolith morphometry and morphology between different congeneric species and populations of the same species from different habitats.