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In 2020–2021, a trial to recruit flat oysters was implemented at a longline farm in the central Adriatic, whereby the efficiency recruitment (n. oyster/dm2) of different suspended substrates was evaluated. Two lantern nets (50 cm diameter; 145 cm h) had different substrates composed of 8 mm wide wrinkled ribbon and empty oyster shells positioned in the upper levels of the lanterns. The tumbling evaluation and the presence of mud were also considered. The efficiency recruitment was similar between the wrinkled ribbon and the oyster shell. Recruitment was in the same proportion on the external rough part of the shells as on the internal smooth part of the shells. No significant differences were shown when comparing the different substrates in terms of recruitment efficiency.

Objective To determine the effect of the Klox fluorescence biomodulation system (Phovia) on the healing of surgical wounds. Study design Prospective, blinded, controlled clinical trial. Sample population Healthy dogs undergoing orthopedic surgery (n = 10). Methods Half of the length of each surgical wound was treated with Phovia, and the remaining 50% was treated with saline solution on the first day after surgery and every 3 days until day 13. Wound healing of treated and control areas within each wound was evaluated via macroscopic assessment and histological and immunohistochemical analysis of treated and control wounds. Results The areas treated with Phovia achieved lower histology scores (P = .001), consistent with complete re-epithelialization, less inflammation of the dermal layer, and greater and more regular deposition of collagen. According to immunohistochemistry, expression of factor VIII, epidural growth factor, decorin, collagen III, and Ki67 was increased in treated compared with untreated tissues. Conclusion Phovia therapy improved re-epithelialization, decreased dermal inflammation, and improved matrix formation in uncomplicated cutaneous incisional wounds by regulating the expression of key biological mediators. Clinical significance Phovia may be a beneficial adjunct to promote the healing of incisional wounds.

Previously, we reported that the administration of a p62/SQSTM1-encoding plasmid demonstrates high safety and signs of clinical benefits for human cancer patients. The treatment also suppressed tumor growth and metastasis in dogs and mouse models. Here we investigated some mechanistic aspects of these effects. In mammary tumors bearing-dogs, i.m. injections of p62 plasmid reduced tumor sizes and their aggressive potential in 5 out of 6 animals, with one carcinoma switching to adenoma. The treatment increased levels of smooth muscle actin in stroma cells and type III collagen in the extracellular matrix, which correlate with a good clinical prognosis. The p62 treatment also increased the abundance of intratumoral T-cells. Because of the role of adaptive immunity cannot be tested in dogs, we compared the protective effects of the p62 plasmid against B16 melanoma in wild type C57BL/6J mice versus their SCID counterpart lacking lymphocytes. The plasmid was only protective in the wild type strain. Also, p62 plasmid amplified the anti-tumor effect of T-cell transfer from tumor-bearing animals to animals challenged with the same tumors. We conclude that the plasmid acts via re-modeling of the tumor microenvironment, making it more favorable for increased anti-cancer immunity. Thus, the p62-encoding plasmid might be a new adjuvant for cancer treatments.

Necropsy of two free-ranging common blackbirds (Turdus merula) found dead in central Italy revealed the presence of a high number of cyclocoelid flukes in the coelomatic cavity. Cyclocoelid flukes primarily infect avian respiratory system. Histologically, air sac walls were covered with a fibrinous exudate containing degenerate heterophils, many trematodes and some colonies of Gram-positive cocci. The superficial bronchi and parabronchi were markedly distended, and the adjacent pulmonary parenchyma was congested and collapsed. Trematodes, surrounded by a mild suppurative to pyogranulomatous inflammatory reaction, were also observed on the pericardial, intestinal, kidney and hepatic serosal surfaces. The death of the two examined birds was likely due to the high parasite load and associated severe lesions. At parasitological examination, flukes showed a tongue-shaped elongate body, tapered anteriorly and rounded posteriorly, of 2,088–2,314 μm in width and 8,268–11,830 μm in length. The mouth was slightly oval and sub-terminal, with a small oral sucker. The oval pharynx measured 250–309 μm, and the two caeca joined posteriorly. Two large (550–702 μm × 450–520 μm) globular testes were situated obliquely to each other, whereas an oval (250 × 300 μm in mean) or round (about 334 μm in diameter) intertesticular ovary was placed in a longitudinal straight line with the testes. The ootype was about 110 μm in diameter, while the brown-yellow eggs measured 131.5 × 73.9 μm in mean. The genital pore was post-pharyngeal, while the symmetrically arranged vitelline glands were not confluent posteriorly. Morphoflogical diagnosis led to the identification of Morishitium polonicum, a cyclocoelid fluke species that typically inhabits the air sacs of blackbirds. The morphological diagnosis was corroborated by molecular phylogenetic analysis of the mitochondrial (CO1, ND1) DNA loci. The present study provides the first report of pathological lesions and death caused by M. polonicum in birds. Keywords: Air sac trematodes, Cyclocoelidae, Death, Histopathological lesions, Morishitium polonicum, Turdus merula

Red mark syndrome (RMS) is a non-lethal inflammatory skin disorder spreading in farmed adult rainbow trout (Oncorhynchus mykiss) and reported worldwide. The aetiology is still uncertain, but positive correlation was found between Midichloria-like organism and RMS-affected fish. Here, we describe the first cases of RMS in Bosnia and Herzegovina. The outbreaks under study occurred in two intensive farms during the late winter and spring of 2020. Affected fish showed signs of disease ascribable to RMS, confirmed by pathological and molecular examination.

Red mark syndrome (RMS) is a skin disorder affecting rainbow trout (Oncorhynchus mykiss). The present work aimed to correlate the gross skin lesions affecting 46 fish sampled from farms surveyed for RMS with their microscopic features, identifying histological parameters that may be suggestive of disease progression. Skin lesions were grossly included in one of three categories (types I, II and III) according to the progressive degree of severity. Histological parameters and anti‐proliferating cell nuclear antigen (PCNA) tissue immunoreactivity were semi‐quantitatively assessed. In the dermis, PCNA‐positive lymphocytes, fibroblasts and endothelial cells were indicative of active phlogosis. A significant increase in PCNA‐immunoreactive lymphocytes, from gross type I to type III cases, was found only in the hypodermis. The histological parameters significantly associated with the gross lesion severity were progressive loss of the epithelium and scales, recruitment of inflammatory cells in the stratum compactum, loss of architecture of the stratum compactum, perivascular and perineural granulomatous inflammation and increase in lymphocyte infiltration of the muscular layer. In the type II and type III categories, inflammation in the hypodermis and muscle displayed a granulomatous pattern, reinforcing the hypothesis of an immunopathological mechanism. The morphological diagnosis of “deep chronic dermatitis associated to panniculitis and myositis, characterised by lympho‐histiocytic and granulomatous reaction” is suggested.

Discoid lupus erythematous (DLE) is a common autoimmune skin disorder of dogs where keratinocytes play a pivotal role both in the innate and adaptive immune responses. As for the innate response, pattern recognition receptors (PRR), including Toll-like receptors (TLRs), can activate macrophages and immune tissue cells allowing for transmission and transduction of signals through cytokine and chemokine release to improve host defenses. In particular, TLR4 can also recognize endogenous molecules such as heat shock proteins produced during reactions to tissue damage. The aim of this study was to evaluate the expression of TLR4, a bacterial lipopolysaccharide sensor, in the skin of dogs with DLE and in normal skin to evaluate a possible involvement of this receptor in the disease pathogenesis. Skin samples of affected dogs had a diffuse and intense expression of TLR4 in the epidermis. Also, the inflammatory infiltrates were immunolabelled. The expression was significantly higher in DLE skin compared to normal skin (**** p &lt, 0.0001). In conclusion, dogs with DLE showed an altered expression of TLR4, which might play an important pathogenic role in the ongoing immunopathologic process, thus being considered a valuable therapeutic potential target for DLE.

Background: In Europe, rainbow trout is one of the main fresh water fish farmed in a constantly developing environment that requires innovative studies to improve farm management, fish welfare and environmental sustainability. The aim of this paper is to investigate the trend of water quality parameters over 10 years, after a feeding strategy change from pellet to extruded feed. Methods: The study was conducted on a farm in central Italy, based on parallel raceways. The cycle started from young rainbow trout (90 &plusmn, 2 g) that were grown until they reached market size. A water sample of 500 cm3 was collected monthly from 2009 to 2019 from the lagoon basin in order to investigate the trends of the total suspended solids (TSS), biochemical oxygen demand (BOD5), chemical oxygen demand (COD), nitrites (NO2-N), nitrates (NO3-N), total ammonia nitrogen (TAN), total phosphorus (TP) and pH. Results: All of the studied parameters (TSS, BOD5, COD, NO2-N, NO3-N, TAN and TP) showed a significant improvement from 2009 to 2019. The pH parameter did not display notable variation during the studied period. The feed conversion ratio (FCR) was also investigated and exhibited a significant improvement from 1.4 to 1.1. Conclusion: Based on the decrease of all the investigated parameters, it is possible to say that extrusion is currently an excellent processing feed technique in aquaculture with a good level of respect for the environment.

Disorders of sexual development (DSDs) are common in horses and cause economic loss in horse breeding. Thus, it is important to develop methods for unambiguous and fast identification of affected horses shortly after birth, as well as those that may propagate the condition to the next generation. Genetic causes of DSDs are multivarious and still little known, and thus development of diagnostic tests requires accumulating knowledge about individual cases and their etiologies. In particular, it is necessary to perform clinical, ultrasound, surgical, histological, cytogenetic, and genetic analyses with close attention in all the affected individuals. This report describes the case of a XX/XY chimeric horse with reproductive apparatus abnormalities and a very low percentage of XY cell in blood highlighting that to avoid undiagnosed case of cell chimeras, above all when studying DSD cases, it is essential to perform both genetic and cytogenetic analyses possibly on more than one tissue.

This article describes a case of acute respiratory distress syndrome in a 3-year-old Uromastyx acanthinura nigriventris . The lizard was presented to the veterinary hospital with an 8-day history of respiratory distress. After the initial physical examination, the patient was treated for the respiratory condition for 3 weeks and subsequently discharged. Unfortunately, the uromastyx died 2 weeks following release from the veterinary hospital, after an episode of acute dyspnea. Acute respiratory syndrome was diagnosed following histological examination of submitted tissue samples. The authors believe the condition was caused by possible environmental exposure to volatile organic compounds.

Previously, we reported that the administration of a p62/SQSTM1-encoding plasmid demonstrates high safety and clinical benefits for human cancer patients, having also suppressed tumor growth and metastasis in dogs and mouse models. Here we investigated the mechanistic aspects of these effects. In mammary tumors bearing-dogs, p62 plasmid i.m. injections reduced tumor volumes, and reverted tumor grade to less aggressive lesions in 5 out of 6 animals, with one carcinoma switching to benign adenoma. The treatment increased levels of alpha-SMA in stroma cells and collagen 3 in the extracellular matrix, both of which correlate with a good clinical prognosis. p62 treatment also increased the abundance of intratumoral T-cell. To test the role of adaptive immunity, we compared protective effects of the plasmid against B16 melanoma in wild type C57BL/6J mice and in the corresponding SCID strain lacking lymphocytes. The plasmid was only protective in the wild type strain. Also, p62 plasmid amplified anti-tumor effect of adoptive T-cell transfer from tumor-bearing animals to animals challenged with the same tumors. We conclude that the plasmid acts indirectly via re-modeling of the tumor microenvironment, making it more favorable for increased anti-cancer immunity. Thus, the p62-encoding plasmid might be a new adjuvant for cancer treatments. Key words: Canine breast tumors, Tumor microenvironment, Anti-cancer immunity, p62 / SQSTM1, DNA plasmid, clinical trial, extracellular matrix.

A necropsy was conducted on a female grey-headed parrot (

Necropsy of two free-ranging common blackbirds (Turdus merula) found dead in central Italy revealed the presence of a high number of cyclocoelid flukes in the coelomatic cavity. Cyclocoelid flukes primarily infect avian respiratory system. Histologically, air sac walls were covered with a fibrinous exudate containing degenerate heterophils, many trematodes and some colonies of Gram-positive cocci. The superficial bronchi and parabronchi were markedly distended, and the adjacent pulmonary parenchyma was congested and collapsed. Trematodes, surrounded by a mild suppurative to pyogranulomatous inflammatory reaction, were also observed on the pericardial, intestinal, kidney and hepatic serosal surfaces. The death of the two examined birds was likely due to the high parasite load and associated severe lesions. At parasitological examination, flukes showed a tongue-shaped elongate body, tapered anteriorly and rounded posteriorly, of 2,088–2,314 μm in width and 8,268–11,830 μm in length. The mouth was slightly oval and sub-terminal, with a small oral sucker. The oval pharynx measured 250–309 μm, and the two caeca joined posteriorly. Two large (550–702 μm × 450–520 μm) globular testes were situated obliquely to each other, whereas an oval (250 × 300 μm in mean) or round (about 334 μm in diameter) intertesticular ovary was placed in a longitudinal straight line with the testes. The ootype was about 110 μm in diameter, while the brown-yellow eggs measured 131.5 × 73.9 μm in mean. The genital pore was post-pharyngeal, while the symmetrically arranged vitelline glands were not confluent posteriorly. Morphoflogical diagnosis led to the identification of Morishitium polonicum, a cyclocoelid fluke species that typically inhabits the air sacs of blackbirds. The morphological diagnosis was corroborated by molecular phylogenetic analysis of the mitochondrial (CO1, ND1) DNA loci. The present study provides the first report of pathological lesions and death caused by M. polonicum in birds., Graphical abstract Image 1, Highlights • Necropsy revealed a high fluke load in the coelom of two deceased common blackbirds. • Based on morphology, the air sac trematode Morishitium polonicum was identified. • Morphological diagnosis was confirmed by molecular analysis. • Blackbird death was likely caused by the high parasite load and severe lesions. • This is the first report of M. polonicum as a cause of death in Turdus merula.

A necropsy was conducted on a female grey-headed parrot (Poicephalus robustus suahelicus) that died following signs of depression, ruffled feathers, and inappetence. Microscopic examination revealed the presence of hemoprotozoa in the liver. A nested polymerase chain reaction (PCR), targeting the mitochondrial cytochrome b gene of Haemoproteus species, Plasmodium species, and Leucocytozoon species, was performed on frozen tissue samples collected at necropsy. The hemoprotozoa were identified by PCR analysis as Leucocytozoon species. Hemoprotozoa are rarely reported in African parrots, and this is the first report of a Leucocytozooon species infection in a Poicephalus robustus suahelicus.

Recent studies highlighted the role of autophagy as a cardinal regulatory system for homeostasis and cancer-related signalling pathways. In this context, the deregulated expression of p62 - Sequestosome1 (p62/SQSTM1) - a protein acting both as an autophagy receptor and signalling hub, has been associated with tumour development and chronic inflammation. Multiple clinical studies test drugs targeting autophagy, and even more research is on the way to clinical trials. However, no comparative investigations have been carried out to identify adequate preclinical models to assess p62-based medicine. In veterinary oncology the role of p62 in cancer-related pathways has been largely ignored. We compared p62 sequences in multiple organisms and found that canine p62 significantly diverges from the humans and from other animals sequences. Then, we chart by immunohistochemistry the expression levels of p62 in canine mammary tumours. A total of 66 tumours and 10 non-neoplastic mammary samples were examined. The expression of p62 was higher in normal tissue and adenomas than carcinomas, with lowest levels of p62 protein detected in high grade carcinomas. In all cases examined the tumour stroma appeared to be p62-negative. Taken together our results would suggest that in dogs the association between p62 expression and cancer cells overturns that reported in human breast carcinoma, where p62 accumulates in malignant cells as compared to normal epithelium. Thus, at least in canine mammary tumours, p62 should be not considered a tumour-rejection antigen for an anti-cancer immunotherapy.

The present work characterized the metabolomic profile of tracheal wash (TW) and exhaled breath condensate (EBC) in healthy horses and horses with respiratory disease. Six asthma-affected horses (group A) and six healthy controls (group H) underwent clinical, endoscopic and cytologic examinations of upper airways to confirm the active phase of asthma. TW and EBC samples were collected from each animal and investigated by proton nuclear magnetic resonance (H-1-NMR) metabolomic analysis. A total of ten out of 38 metabolites found in the TW were significantly different between the groups (p < 0.05). Higher concentrations of histamine and oxidant agents, such as glutamate, valine, leucine and isoleucine, as well as lower levels of ascorbate, methylamine, dimethylamine and O-phosphocholine, were found in group A compared to group H. Eight metabolites were found in equine EBC, namely methanol, ethanol, formate, trimethylamine, acetone, acetate, lactate and butanone, previously observed also in human EBC. Despite the fact that this was a pilot study, the results showed that the metabolomic analysis of TW and EBC has the potentiality to serve as a basis for diagnostic tools in horses with asthma.

Necropsies were conducted on a female blue-fronted Amazon (Amazona aestiva) and a female yellow-headed Amazon (Amazona oratrix) that died after depression, ruffled feathers, diarrhea, and biliverdin in the urine. Gross and microscopic examinations revealed multifocal necrosis in the liver, spleen, lungs, kidneys, intestines, and heart caused by acute bacteremia. Yersinia pseudotuberculosis, serogroup O:1a, was isolated by culturing from the visceral lesions in the liver, intestines, and spleen. Virulence gene analysis showed the presence of the inv gene and the complete pathogenicity island: IS100, psn, yptE, irp1, irp2 ybtP-ybtQ, ybtX-ybtS, and int asnT-Int. Histopathologic findings and chemical analysis also demonstrated hepatic hemosiderosis. As has been demonstrated in other species, hemosiderosis may predispose Amazona spp. to systemic infection with Y. pseudotuberculosis after enteric disease.

Background Chronic intestinal pseudo-obstruction (CIPO) is a rare clinical syndrome in veterinary medicine characterized by severe intestinal dysmotility without evidence of mechanical occlusion of the intestinal lumen. The exact pathogenesis of CIPO is unknown. Case presentation A 1-year-old male Miniature Bull Terrier dog was presented with a history of chronic weight loss, regurgitation, lethargy, vomiting and diarrhea. The dog was submitted for exploratory laparotomy. A full thickness intestinal biopsy was taken and a CIPO was suspected. The clinical condition deteriorated and the dog was euthanized. At gross examination the small intestine was severely dilated. Histologically severe fibrosis of the submucosa and severe atrophy of the tunica muscularis were present in small intestine and colon. Immunohistochemical examination with a panel of antibodies for gastro-intestinal neuromuscular disease-associated antigens revealed a severely reduced expression of alpha-smooth muscle actin in the tunica muscularis. Conclusions This case report describes the gross, histological and immunohistochemical findings of CIPO affecting a 1-year-old Miniature Bull Terrier; on the basis of these findings a myopathic form of CIPO is hypothesized in this case.

A pregrowing trial was carried out using nursery-stage cupped oyster (Crassostrea gigas) in order to evaluate weight, length, mortality, and final costs of spat grown in long-line aquaculture at different sampling times (t1–t5) over 81 days. Spat were reared at two stocking densities in polyvinyl chloride cylinders (10 cm diameter, 24 cm length) placed within lanterns composed of five levels (60 cm diameter, 11 cm height). Each lantern level hosted two cylinders. Lanterns were hung 6 m below the surface of the water in the Middle Adriatic Sea. A total of 53, 100 spat (weight 0.033 g, length 5 mm) were divided into seven cylinders at either high density (H ; 5055 oysters/cylinder, 167.3 g/cylinder) or low density (L ; 2530 oysters/cylinder, 83.7 g/cylinder). On the basis of the ANOVA analysis, spat were significantly heavier (0.352 vs. 0.227 g ; P < 0.001) and longer (12.22 vs. 10.47mm ; P < 0.001) in the L group compared to the H group. Considering the time factor, oysters were significantly heavier (0.941 g ; P < 0.001) and longer (21.30 mm ; P < 0.001) at t5 than at any previous sampling time (t1–t4). A higher rate of mortality was initially observed in the L group (6.7% at t1) ; however, mortality in the H group was higher at later sampling dates, reaching the highest level at t4 (32.4%). Spat grew better at the low stocking density, demonstrating that a higher stocking density is not suitable.

Visna/Maedi virus (VMV), a small ruminant lentivirus responsible for lymphoproliferative pneumonia, encephalitis, arthritis and/or mastitis in sheep, has been detected in different non-lymphoid organs. However, only a few investigations have been carried out in lymphoid tissues. In this study, some lymphoid tissues and lymph node draining or non-draining VMV target organs from five sheep infected experimentally by the respiratory route three years previously were investigated. Archival samples of spleen, red bone marrow, caudal mediastinal lymph nodes, mammary lymph nodes, popliteal lymph nodes and mesenteric lymph nodes were tested by PCR for the presence of proviral DNA. Popliteal and mesenteric lymph node samples were tested also by immunohistochemi - cal staining of the viral capsid antigen p28. The proviral DNA was detected by PCR in all the lymphoid tissue samples from the infected sheep. The viral antigen was stained in mononuclear cells in popliteal and mesenteric lymph nodes of the infected sheep. Although the lymph nodes draining the classical target organs seem to be more infected than the others, both the viral capsid antigen and the proviral DNA were present also in lymph nodes draining non-target organs, such as the mesenteric lymph nodes. These findings show the presence of VMV in different lymphoid tissues in the late stages of infection and suggest a potential role of these tissues as a site for viral reservoir and replication, even three years after infection.

Red mark syndrome (RMS) and US strawberry disease (US SD) are skin disorders affecting rainbow trout farmed in Europe and USA. The disease etiology has not yet been established. In spite of specific investigations, identifying Rickettsia-like organism (RLO)- and Midichloria-like organism (MLO)-related DNA in affected individuals, these pathogens have never been observed. We performed histological, ultrastructural and biomolecular analysis on skin and spleen samples of trout with RMS. Examination by TEM revealed the presence of intracytoplasmic microorganisms resembling Rickettsiales within macrophages, fibroblasts and erythrocytes. The microorganisms were oval or short rod shaped (400–800 nm in length and 100–200 nm in width) and often showed a cell wall similar to Gram-negative bacteria. PCR analysis for Rickettsiales supported these findings: 53% of affected trout were positive by both PCR and TEM The primers RiFCfw-RiFCrev were used to anneal both the RLO 16S DNA sequence and the MLO 16S DNA sequence. For this reason, and in agreement with previous studies confirming the presence of Rickettsiales-related DNA in trout with RMS, we assume that TEM detected microorganisms morphologically consistent with bacteria belonging to Rickettsiales order and could be considered as possible causative agents of RMS.

A case of an ovarian tumor is reported in an 8-yr-old South American sea lion (Otaria flavescens) kept in a marine park in Malta (35.57 degrees N, 14.25 degrees E). The neoplasm was a solid mass of dense sheets and nests of round to polyhedral, irregularly shaped cells with abundant, finely vacuolated cytoplasm. The nuclei were uniformly small and round to oval. The supporting stroma contained thecal cells. The tumor cells were positive for positive inhibin and vimentin and focally positive for cytokeratin by immunohistochemistry. The neoplasm was diagnosed as an ovarian sex cord-stromal tumor, specifically an interstitial cell tumor.

The aim of this study was to investigate the carbohydrate composition of mucosubstances in the equine guttural pouches using conventional histochemical tests in conjunction with glycolytic digestion to degrade different classes of glycosoaminoglycans. In the goblet cells, the mucopolysaccharides contained chondroitin sulfate B, heparin, heparan sulfate and sialic acid residues. The acinar cells also expressed these substances (except for heparin), whereas the ductal cells produced chondroitin sulfate B and sialic acid. Neutral sugars were also found in each cell type. The diversity of the glycocomponents found in the auditory tube suggests that they may have important functional roles. Indeed, the glycosoaminoglycans provide a hydrophilic environment that prevents dehydration and desiccation of the guttural membranes during air passage. Additionally, these glycomolecules may be involved in the pathogenesis of some bacterial disease in horses, such as equine strangles.

Sheep develop clinical diseases after 3-5 years after infection with Mycobacterium avium subsp. paratuberculosis (MAP). Clinical symptoms of paratuberculosis include persistent diarrhea and weight loss due to a chronic inflammation of the small intestine. Tissue alterations in the areas of the ileo-cecal junction are often observed. Here, we investigate the molecular processes underlying tissue damages in intestinal mucosa of 14 sheep showing either tuberculoid or lepromatous form of MAP enteritis. We found that E-cadherins, alpha-catenin and beta1-integrins were present at significant low levels in tissues of sheep affected by lepromatous form and that this pattern was associated with high expression of TGF-beta, IL-10, IL-1beta, and TNF-alpha and with a modest increase of CD4+ and CD25+ T cells. Tissues of sheep with the tuberculoid form showed high expression of IFNgamma, IL-12, and MCP-1 and a significant presence of CD4+ and CD25+ T cells. Finally, anti-transglutaminase (tTG) IgG1 antibodies were detected in sera of infected animal belonging to both groups, as already described for human inflammatory bowel diseases. Our results further stress the similarities in the clinical and histological features between ruminant paratuberculosis and human intestinal inflammatory diseases.

The aims of the study were to evaluate a new PCR protocol designed to detect Aeromonas salmonicida in fish tissues and to develop a non-destructive method for the diagnosis of furunculosis. A set of primers (Fer3, Fer4), flanking a fragment of the fstA gene (coding for the ferric-siderophore receptor) was designed, showing to be sensitive and specific. When compared to PCR methods previously reported, the new protocol recognized all the 69 A. salmonicida strains evaluated, with no cross-reactions with the other bacterial species analysed. Sensitivity assays were performed in fish tissues seeded with serial dilutions of pure cultures of A. salmonicida and mixed cultures of this bacterium with Vibrio anguillarum and Aeromonas hydrophila. Detection limits obtained were of 60 and 450 bacterial cells 100 mg(-1) of tissue, respectively. Mucus and blood were evaluated in order to develop a non-destructive tool to detect the pathogen. The detection limits in seeded mucus and blood samples were 2.5 x 10(2) and 1 x 10(5) bacterial cells mL(-1), respectively. When the method was used to detect A. salmonicida in asymptomatic wild salmon, four samples of mucus and six of blood were positive, corresponding to 6 out of the 31 fish examined, whereas only one of the samples resulted positive by culture methods. It is concluded that the PCR protocol evaluated is fast, specific and sensitive to detect A. salmonicida in infected and asymptomatic fish, and will be helpful for the control of the disease through the prompt detection of carriers within fish populations.

A spontaneous intestinal adenocarcinoma in a 3-year-old blue gularis Fundulopanchax sjostedti with a history of swelling of the trunk and anorexia is described. At necropsy, there was a large, firm, irregular mass originated from the intestine and distending the abdominal cavity. Histologically the intestinal mass showed proliferation of neoplastic epithelial cells arranged in irregularly shaped islands and dense sheets with papillotubular ingrowths. Immunohistochemically the neoplastic cells expressed cytokeratin AE1/AE3, PCNA and interestingly the mutant p53 protein. On the basis of the histopathological and immunohistochemical features, an intestinal adenocarcinoma was diagnosed. The expression of the mutant p53 protein in tumour cells may demonstrate its role in neoplastic transformation of enterocytes as observed in human cancers.

INTRODUCTION: CD25 (IL-2Ra) is the α-subunit of the membrane-bound interleukin-2 receptor (IL-2R), which is mainly expressed by regulatory T cells (Tregs). Tregs play a pivotal role in the homeostasis of the immune system and in the modulation of the immune response (1,2). Two type of Tregs are known: natural Treg and adaptive Treg. Normally these cells regulate immune system activity and prevent autoimmunity. Imbalanced function or number of these cells, either enhanced or decreased, might lead to tumor development and autoimmunity, respectively (1,2,3,4). In the present study we hypothesized the presence of CD25+ cells among tumor infiltrating lymphocytes (TILs) that are normally associated with feline injection-site sarcoma (FISS), a type of tumor considered a good model to study a link between inflammation and neoplasia. To this end the CD25 protein expression levels were analyzed by immunohistochemistry in eighteen cases of FISS and in 15 cases of post-injection panniculitis (PIP).

The work aimed to determine the welfare status of rainbow trout (50±15 g), intensively reared to commercial size in twoconcrete raceways (V1, V2) at different stocking densities, to reach final load of 40 kg/m3 and 20 kg/m3, respectively.Throughout the out-phase, plasma parameters and hepatic glycogen content were determined every three months andcompared with those of rainbow trout of the same age and mean weight, reared extensively (VE). At the end of the trialsignificant differences of the monitored parameters were found between rainbow trout reared in raceways (V1, V2) andthose held in the reservoir (VE). Plasma triglycerids, total cholesterol and alkaline phosphatase in fish reared intensivelywere significantly different from fish reared in extensive conditions. The highest levels of these parameters in V1 andV2 could be justified mostly by the administration of balanced feeding and consequent modifications of energeticmetabolism and in small part to the high density of the intensive rearing conditions. Transaminases (AST, ALT) increasedin V1 and V2 only at the first sampling. Afterwards, the activity of these two enzymes returned to the normal range atthe subsequent assays, suggesting a capacity of the rainbow trout to adapt to the severe conditions of confinement, typicalof out-basins. No significant differences among groups were observed for total protein, glucose, CK and LDH. A similarsituation was also shown for liver glycogen content.