Slightly viscous dispersions of mucoadhesive polymers as vehicles for nasal administration of dopamine and grape seed extract-loaded solid lipid nanoparticles.

With the aim to find an alternative vehicle to the most used thermosensitive hydrogels for efficient nanotechnology-based nose-to-brain delivery approach for Parkinson's disease (PD) treatment, in this work we evaluated the Dopamine (DA) and the antioxidant grape seed-derived pro-anthocyanidins (Grape Seed Extract, GSE) co-loaded solid lipid nanoparticles (SLNs) put in slight viscous dispersions (SVDs). These SVDs were prepared by dispersion in water at low concentrations of mucoadhesive polymers to which SLN pellets were added. For the purpose, we investigated two polymeric blends, namely Poloxamer/Carbopol (PF-127/Carb) and oxidized alginate/Hydroxypropylmethyl cellulose (AlgOX/HPMC). Rheological studies showed that the two fluids possess Newtonian behaviour with a viscosity slightly higher that water. The pH values of the SVDs were mainly within the normal range of nasal fluid as well as almost no osmotic effect was associated to both SVDs. All the SVDs were capable to provide DA permeation through nasal porcine mucosa. Moreover, it was found that PF-127/Carb blend possesses penetration enhancer capability better than the Alg OX/HPMC combination. Flow cytometry studies demonstrated the uptake of viscous liquids incorporating fluorescent SLNs by human nasal RPMI 2650 cell in time-dependent manner. In conclusion, the SVD formulations may be considered promising alternatives to thermosensitive hydrogels strategy. Moreover, in a broader perspective, such SVD formulations may be also hopeful for treating various neurological diseases beyond PD treatment.

Relationship between Fat Status, Stage of Gonadal Maturity and Hormonal Variation of Turdus philomelos (C.L. Brehm, 1831) Wintering in Apulia during 2018-2020.

In this study, we aimed to evaluate the development of the fattening condition and the reproductive status of the song thrush from December to February. For this purpose, the chemical and fatty acid compositions of the pectoral muscle were analyzed in relation to the fattening state of the birds. Moreover, their reproductive activity was evaluated via the anatomical and pathological examination of tissues and through the assessment of sex steroid profiles. One hundred ninety-five thrushes captured by local hunters during the 2018-2019 and 2019-2020 hunting seasons in different provinces of the Apulia region in Italy were used. The first step was the measurement of bird body mass, and the amount of subcutaneous body fat was estimated visually. During post-mortem examinations, the pectoral muscle was excised and used for chemical and fatty acid analysis and a hormone assay, respectively. Moreover, ovaries and testicles were evaluated to determine the degree of maturation and thus the reproductive status of the birds. The results regarding fattening status and fatty acid profile confirmed that in January-February, thrushes change their diet, increasing their intake of oleic acid, likely to better cope with low temperatures and prepare for long-distance migration. In both male and female thrushes, the concentrations of sex hormones confirmed a phase of reproductive quiescence from December to February, which was also confirmed through histological examination of the gonads.

Effect of rearing system (free-range vs cage) on gut and muscle histomorphology and microbial loads of Italian White breed rabbits.

OBJECTIVE: The growing consumers' interest on animal welfare has raised the request of products obtained by alternative rearing systems. The present study was conducted to assess the influence of housing system on gut and muscle morphology and on microbial load in rabbits reared under free-range (FR) and cage system (CS). METHODS: A total of forty weaned (35 days of age) male Italian White breed rabbits were allotted according to the rearing system, and at 91 days of age were randomly selected and slaughtered for the morphological evaluation of tissue from duodenum and longissimus lumborum. Morphometric analysis of the villus height, villus width, crypt depth, villus height/crypt depth ratio, and villus surface was performed. The microbial loads on hind muscle was determined by total mesophilic aerobic count (TMAC), Escherichia coli and Enterobacteriaceae; whereas, total anaerobic bacteria count (TABC) and TMAC, E. coli and Enterobacteriaceae was determined on caecal content. RESULTS: Rearing system did not interfere with the duodenum and muscle histomorphology in both rabbit groups. Similarly, microbial load of caecal content showed no significant differences on the TABC and TMAC. Conversely, significant difference was found for E. coli strains in caecal content, with the lower counts in FR compared to CS rabbits (p<0.01). Microbiological assay of muscle revealed significant lower TMAC in FR vs CS rabbits (p< 0.05). All rabbit meat samples were negative for E. Coli and Enterobacteriaceae. CONCLUSION: Free-range could be considered a possible alternative and sustainable rearing system in rabbits to preserve gut environment and muscle quality.

Immunohistochemical, pharmacovigilance, and omics analyses reveal the involvement of ATP-sensitive K+ channel subunits in cancers: role in drug-disease interactions.

Background: ATP-sensitive-K+ channels (KATP) are involved in diseases, but their role in cancer is poorly described. Pituitary macroadenoma has been observed in Cantu' syndrome (C.S.), which is associated with the gain-of-function mutations of the ABCC9 and KCNJ8 genes. We tested the role of the ABCC8/Sur1, ABCC9/Sur2A/B, KCNJ11/Kir6.2, and KCNJ8/Kir6.1 genes experimentally in a minoxidil-induced renal tumor in male rats and in the female canine breast cancer, a spontaneous animal model of disease, and in the pharmacovigilance and omics databases. Methods: We performed biopsies from renal tissues of male rats (N = 5) following a sub-chronic high dosing topical administration of minoxidil (0.777-77.7 mg/kg/day) and from breast tissues of female dogs for diagnosis (N = 23) that were analyzed by immunohistochemistry. Pharmacovigilance and omics data were extracted from EudraVigilance and omics databases, respectively. Results: An elevated immunohistochemical reactivity to Sur2A-mAb was detected in the cytosol of the Ki67+/G3 cells other than in the surface membrane in the minoxidil-induced renal tumor and the breast tumor samples. KCNJ11, KCNJ8, and ABCC9 genes are upregulated in cancers but ABCC8 is downregulated. The Kir6.2-Sur2A/B-channel opener minoxidil showed 23 case reports of breast cancer and one case of ovarian cancer in line with omics data reporting, respectively, and the negative and positive prognostic roles of the ABCC9 gene in these cancers. Sulfonylureas and glinides blocking the pancreatic Kir6.2-Sur1 subunits showed a higher risk for pancreatic cancer in line with the positive prognostic role of the ABCC8 gene but low risks for common cancers. Glibenclamide, repaglinide, and glimepiride show a lower cancer risk within the KATP channel blockers. The Kir6.2-Sur1 opener diazoxide shows no cancer reactions. Conclusion: An elevated expression of the Sur2A subunit was found in proliferating cells in two animal models of cancer. Immunohistochemistry/omics/pharmacovigilance data reveal the role of the Kir6.1/2-Sur2A/B subunits as a drug target in breast/renal cancers and in C.S.

Combined Dopamine and Grape Seed Extract-Loaded Solid Lipid Nanoparticles: Nasal Mucosa Permeation, and Uptake by Olfactory Ensheathing Cells and Neuronal SH-SY5Y Cells.

We have already formulated solid lipid nanoparticles (SLNs) in which the combination of the neurotransmitter dopamine (DA) and the antioxidant grape-seed-derived proanthocyanidins (grape seed extract, GSE) was supposed to be favorable for Parkinson's disease (PD) treatment. In fact, GSE supply would reduce the PD-related oxidative stress in a synergic effect with DA. Herein, two different methods of DA/GSE loading were studied, namely, coadministration in the aqueous phase of DA and GSE, and the other approach consisting of a physical adsorption of GSE onto preformed DA containing SLNs. Mean diameter of DA coencapsulating GSE SLNs was 187 ± 4 nm vs. 287 ± 15 nm of GSE adsorbing DA-SLNs. TEM microphotographs evidenced low-contrast spheroidal particles, irrespective of the SLN type. Moreover, Franz diffusion cell experiments confirmed the permeation of DA from both SLNs through the porcine nasal mucosa. Furthermore, fluorescent SLNs also underwent cell-uptake studies by using flow cytometry in olfactory ensheathing cells and neuronal SH-SY5Y cells, evidencing higher uptake when GSE was coencapsulated rather than adsorbed onto the particles.

Thoracic and Abdominal Mesothelioma in an Older Horse in Lazio Region.

A Quarter Horse, a gelding aged 22, was subjected to a clinical examination for colic syndrome. During admission to the clinic, blood counts and ultrasound examination were performed. Ultrasound revealed abdominal masses and abundant accumulation of pleural (50 L) and abdominal fluid (100 L). Cytology was performed on the aspirated fluid. The patient was euthanized. The autopsy examination revealed abundant effusion and nodular masses on the peritoneum, omentum, lungs, heart, and mediastinum. A diagnosis of epithelioid mesothelioma was made via histopathology and confirmed with immunohistochemistry; it showed positive antibodies against cytokeratin (CK) and vimentin. Mesothelioma is a rare cancer in older horses. It is important to employ the correct differential diagnostics using the available methods, providing valid ante-mortem support to the clinical veterinarian and monitoring the territory using this species as a valid biological indicator.

Ex-Vivo Evaluation of "First Tip Closing" Radiofrequency Vessel Sealing Devices for Swine Small Intestinal Transection.

This study compared burst pressure (BP), number of activations, and histological assessment of ex vivo swine small intestine loops transected by stapler, a single fulcrum radiofrequency vessel sealing (RFVS) device, and the newly-developed jaws RFVS. Fifty (n = 50) 20 cm long jejunal loops were randomly assigned to be transected with RFVS devices and linear stapler (Caiman5, Caiman Maryland, Caiman12, Ligasure Atlas, and Stapler group as control respectively). Caiman5, Caiman12 and stapler required only one activation to complete the sealing. The mean BP in Caiman5 and Caiman Maryland groups were significantly lower (p < 0.05) than the S group as control and the other RFVS devices studied. RFVS Caiman12 and Ligasure Atlas produced mean BP values that were close to the Control and did not differ between them. The lumen was totally closed in the Caiman12 and Ligasure Atlas groups. The findings of this investigation were promising; we discovered that Caiman12 and Ligasure Atlas produce comparable mechanical capabilities as well as stapled intestinal closure, however Caiman12 need a single activation to complete the transection.

Consequences of SUR2[A478V] Mutation in Skeletal Muscle of Murine Model of Cantu Syndrome.

(1) Background: Cantu syndrome (CS) arises from gain-of-function (GOF) mutations in the ABCC9 and KCNJ8 genes, which encode ATP-sensitive K+ (KATP) channel subunits SUR2 and Kir6.1, respectively. Most CS patients have mutations in SUR2, the major component of skeletal muscle KATP, but the consequences of SUR2 GOF in skeletal muscle are unknown. (2) Methods: We performed in vivo and ex vivo characterization of skeletal muscle in heterozygous SUR2[A478V] (SUR2wt/AV) and homozygous SUR2[A478V] (SUR2AV/AV) CS mice. (3) Results: In SUR2wt/AV and SUR2AV/AV mice, forelimb strength and diaphragm amplitude movement were reduced; muscle echodensity was enhanced. KATP channel currents recorded in Flexor digitorum brevis fibers showed reduced MgATP-sensitivity in SUR2wt/AV, dramatically so in SUR2AV/AV mice; IC50 for MgATP inhibition of KATP currents were 1.9 ± 0.5 × 10-5 M in SUR2wt/AV and 8.6 ± 0.4 × 10-6 M in WT mice and was not measurable in SUR2AV/AV. A slight rightward shift of sensitivity to inhibition by glibenclamide was detected in SUR2AV/AV mice. Histopathological and qPCR analysis revealed atrophy of soleus and tibialis anterior muscles and up-regulation of atrogin-1 and MuRF1 mRNA in CS mice. (4) Conclusions: SUR2[A478V] "knock-in" mutation in mice impairs KATP channel modulation by MgATP, markedly so in SUR2AV/AV, with atrophy and non-inflammatory edema in different skeletal muscle phenotypes.

Dirofilarioses in two cats in southern Italy.

Two cats infected by Dirofilaria immitis and Dirofilaria repens, respectively, were taken to two different private practitioners for a clinical examination. The analyses conducted on the first cat revealed a microfilaraemia due to D. repens of 66 mfs/mL by a modified Knott's test. No clinical signs of D. repens infection were observed in the cat. The animal was euthanised because of a lymphoma condition, and two adult females of D. repens were found in the subcutaneous tissue of the lumbar and left scapular regions at the post-mortem examination. The second cat showed severe abnormalities in the white blood cells, including eosinophil count. Microfilariae of D. immitis were detected in the blood smear, with an average length (n = 2) of 296.2 µm. These clinical cases represent the first reports of feline dirofilarioses in southern Italy and are indicative of a common occurrence of dirofilarial infection in the local canine population.

Absorbable fixation straps for laparoscopic gastropexy in dogs.

OBJECTIVE: To evaluate the feasibility and efficacy of laparoscopic absorbable fixation straps (AFS) for laparoscopic gastropexy in dogs. STUDY DESIGN: Cadaveric and prospective clinical study. ANIMALS: Five dog cadavers for the cadaveric study; 12 dogs for the clinical study. METHODS: The pyloric antrum was affixed to the abdominal wall laparoscopically by applying a series of straps. The cadaveric study assessed potential challenges during the procedure and stomach mucosal penetration. For the clinical study, the total duration of surgery, time to complete the gastropexy, and the number of straps used were recorded. Ultrasound evidence of adhesion, complications, and weight were monitored at 7, 30, and 90 days after surgery. Owner satisfaction was evaluated at the 6-month follow-up. RESULTS: The total duration of surgery was 25.8 minutes (range, 19-39; SD, 6.7), and the time to complete the gastropexy was 10.1 minutes (range, 7-19; SD, 3.9). The linear regression analysis revealed an inverse correlation between the time to complete the gastropexy and the order of the surgeries (r2  = 0.75, P < .05). No complications were recorded. Ultrasound examination was used to confirm gastropexy at all follow-ups. CONCLUSION: Laparoscopic gastropexy with AFS was performed in both cadavers and clinical animals with minimal complications. Persistent adhesion was demonstrated during ultrasound evaluations and in one postmortem evaluation. CLINICAL SIGNIFICANCE: This novel laparoscopic technique can be employed safely, effectively, and reasonably quickly, and the learning curve is expected to be relatively short.

Microvascular Density, Endothelial Area, and Ki-67 Proliferative Index Correlate Each Other in Cat Post-Injection Fibrosarcoma.

Soft tissue sarcomas are a large group of different tumor types both in humans and in animals. Among them, fibrosarcoma is the most frequent malignant mesenchymal tumoral form in cats, representing up to 28% of all cat skin tumors, while human fibrosarcoma, fortunately, only represents 5% of all sarcomas and 0.025% of the world-wide burden of tumors. This low incidence in humans leads to consideration of this group of tumoral diseases as rare, so therapeutic options are few due to the difficulty of starting clinical trials. In this context, the identification of research models for fibrosarcomas could be of great interest to deepen knowledge in this field and recognize new or possible biological pathways involved in tumor progression and metastasis. Angiogenesis is considered a fundamental scattering cause of tumor aggressiveness and progression in all forms of cancer, but only a few research parameters were developed and reported to express them quantitatively and qualitatively. The role in angiogenesis of microenvironmental stromal cells, such as fibroblasts, lymphocytes, mast cells, and macrophages, was largely demonstrated since this topic was first approached, while quantification of new vessels and their blood capacity in tumoral area is a relatively recent approach that could be well developed thanks to expertise in immunohistochemistry and image analysis. In this paper, a crossing study evaluating microvascular density (MVD), endothelial area (EA), and Ki-67 proliferative index was reported for a series of formalin-fixed and paraffin-embedded tissue samples from 99 cat patients, affected by cat post-injection fibrosarcoma, by using a till ×400 magnification light microscopy. We aim to demonstrate that cat pets may be considered a useful animal model for better studying the correspondent human diseases and we report, for the first time to our knowledge, experimental data in terms of correlation among MVD, EA, and Ki-67 strictly involved in aggressiveness and tumoral progression.

Systematic Pathologic Findings Report of Callosciurus finlaysonii (Horsfield, 1823) (Rodentia, Sciuridae) Squirrels from Maratea area (South Italy) to Investigate Species-Specific Pathologies, Reliability of CO2 Euthanasia Method, and Possible Use as Environmental Sentinels.

The aim of the present study was to macroscopically and microscopically describe the main pathological findings occurring in this invasive species, in order to better understand the real risks for naïve animals and humans. The present study was conducted on Callosciurusfinlaysonii squirrels (n = 165), captured in the Maratea area and euthanatized with CO2 according to a population control of invasive species of the Italian Agriculture Ministry project (ex CIPE project) and conferred to the Department of Veterinary Medicine of Bari (Italy). Macroscopic analysis demonstrated heart, lung, and liver congestion, probably due to the euthanasia method, and variable lesions of bowel, liver, and kidney. The microscopically examination showed the presence of lymphocytic infiltration in the lower layers of the bowel, suggesting enteritis. To the best of our knowledge, this is the first systemic report of gross and microscopical anatomopathological lesions in C.finlaysonii, in South Italy. The results could be useful to fill a gap of knowledge of this species in Italy.

Clinical, haematological and biochemical findings in tigers infected by Leishmania infantum.

BACKGROUND: A large number of animal species are susceptible to Leishmania infantum (Kinetoplastida, Trypanosomatidae) in endemic areas, including domestic and wild felids such as tigers (Panthera tigris). Knowledge on the infection of this endangered species is still at its infancy, and therefore this study aims to identify clinical presentation and clinicopathological findings of tigers naturally infected by L. infantum. RESULTS: Tigers either L. infantum-positive (group A) or -negative (group B) were apparently healthy or presented visceral leishmaniasis unrelated conditions, except for one animal in which a large non-healing cutaneous lesion was observed. However, histological exam and immunohistochemistry carried out on the lesion excluded the presence of L. infantum amastigotes. Biochemical analysis showed that the average concentration of total proteins, globulins and haptoglobin were significantly higher (p < 0.01, p = 0.01 and p = 0.02, respectively), while the albumin/globulin ratio significantly lower (p = 0.05) in group A compared with group B. The biochemical alterations were partially confirmed by the serum protein electrophoresis results revealing a significant increase in the total protein value (p = 0.01) and hypergammaglobulinemia (p = 0.03) but an unmodified albumin/globulin ratio in group A. CONCLUSIONS: In this study tigers infected by L. infantum have shown to be mainly asymptomatic. The absence of clinical signs may lead veterinarians to overlook leishmaniasis in animals kept in captivity. Therefore, diagnostic and screening tests as serology should be part of routinely surveillance programs to be performed on tigers in zoological gardens located in endemic areas. Though only few protein-related laboratory abnormalities were recorded in infected animals, they could provide diagnostic clues for a first suspicion of L. infantum infection in tigers. Indeed, considering the high risk of zoonotic transmission in heavily frequented environment as zoos, a prompt diagnosis of L. infantum infection is of pivotal importance.

The hydroxypropyl-ß-cyclodextrin-minoxidil inclusion complex improves the cardiovascular and proliferative adverse effects of minoxidil in male rats: Implications in the treatment of alopecia.

The efficacy of minoxidil (MXD) ethanolic solutions (1%-5% w/v) in the treatment of androgenetic alopecia is limited by adverse reactions. The toxicological effects of repeated topical applications of escalating dose (0.035%-3.5% w/v) and of single and twice daily doses (3.5% w/v) of a novel hydroxypropyl-ß-cyclodextrin MXD GEL formulation (MXD/HP-ß-CD) and a MXD solution were investigated in male rats. The cardiovascular effects were evaluated by telemetric monitoring of ECG and arterial pressure in free-moving rats. Ultrasonographic evaluation of cardiac morphology and function, and histopathological and biochemical analysis of the tissues, were performed. A pharmacovigilance investigation was undertaken using the EudraVigilance database for the evaluation of the potential cancer-related effects of topical MXD. Following the application of repeated escalating doses of MXD solution, cardiac hypertrophy, hypotension, enhanced serum natriuretic peptides and K+ -ion levels, serum liver biomarkers, and histological lesions including renal cancer were observed. In addition, the administration of a twice daily dose of MXD solution, at SF rat vs human = 311, caused reductions in the systolic, diastolic, and mean blood pressure of the rats (-30.76 ± 3%, -28.84 ± 4%, and -30.66 ± 5%, respectively, vs the baseline; t test P < .05). These effects were not reversible following washout of the MXD solution. Retrospective investigation showed 32 cases of cancer associated with the use of topical MXD in humans. The rats treated with MXD HP-ß-CD were less severely affected. MXD causes proliferative adverse effects. The MXD HP-ß-CD inclusion complex reduces these adverse effects.

Effect of incremental levels of sumac (Rhus coriaria L.) seed powder on growth, carcass traits, blood parameters, immune system and selected ileal microorganisms of broilers.

Sumac (Rhus coriaria L.) is a plant species belong to Anacardiaceous family that is worldwide diffused. The sumac seed power (SSP), produced by grinding dried fruits, is recognized to have defensive and beneficial effects on numerous health­related problems. In this study, SSP was included in broilers basal­diet to investigate the comparative effects of different levels of SSP on performance, carcass characteristics, blood parameters, immune system and ileal microorganisms. A total of 225, one day­old male broilers (Ross 308) were randomly assigned to the five dietary treatments with three replicates per treatment. The experimental diets were: basal­diet (BD); and BD including 0.05, 0.10, 0.15 and 0.20% SSP, respectively. During the whole feeding period (42 days), birds fed corn­based grower (1­21 days) and finisher (22­42 days) diets, respectively. Results indicated that supplementing SSP had no effect on broiler body weight gain, feed intake and feed conversion as well as carcass characteristics (P > 0.05). Similarly, blood total protein, albumin, glucose and triglyceride were not influenced by dietary SSP. Conversely, serum total cholesterol and LDL­cholesterol levels were decreased, while HDL­cholesterol increased in all SSP fed groups compared to control (P < 0.05). In this study the addition of SSP in broilers diets did not show any effect on blood heterophils and lymphocyte. Moreover, the lactobacillus count remained unaffected by dietary treatments, while E. coli count in broiler ileal content was lower when fed 0.10% SSP than the other groups (P < 0.05). Thus, the present findings indicated a positive effect of feeding SSP (especially at 0.10% diet) on blood cholesterol levels and E. coli count in broiler chickens

Effect of cat litters on feline coronavirus infection of cell culture and cats.

OBJECTIVES: Feline infectious peritonitis (FIP) is caused by infection with feline coronavirus (FCoV). FCoV is incredibly contagious and transmission is via the faecal-oral route. FCoV infection, and therefore FIP, is most common in breeder and rescue catteries, where many cats are kept indoors, using litter trays. Whether it is possible to break the cycle of FCoV infection and reinfection using cat litters has never been investigated. The aim of the study was to examine the effect of cat litters on FCoV infectivity and virus load in multi-cat households, and transmission frequency. METHODS: Fifteen cat litters were mixed and incubated with FCoV, centrifuged and the supernatants tested in vitro for the ability to prevent virus infection of cell culture. To test applicability of in vitro results to real life, virus load was measured in two households in a double crossover study of four Fuller's earth-based cat litters by testing rectal swabs using FCoV reverse transcriptase quantitative PCR. RESULTS: Four litters abrogated FCoV infection of cell culture, nine reduced it to a greater or lesser extent and two had no effect. One brand had different virus inhibitory properties depending on where it was manufactured. Fuller's earth-based litters performed best, presumably by adsorbing virus. In the field study, there appeared to be less virus shedding on one Fuller's earth-based cat litter. CONCLUSIONS AND RELEVANCE: The in vitro study successfully identified cat litters that inactivate FCoV; such litters exist so do not need to be developed. Fuller's earth-based litters best prevented infection of cell culture, but did not completely abrogate FCoV transmission in two multi-cat households. A dust-free clumping Fuller's earth litter appeared to fare best, but virus shedding also varied on the control litters, complicating interpretation. Sawdust-based cat litters are not useful in FCoV-endemic households because they track badly and have a poor effect on virus infection.

Histological features of Rickettsia-like organisms in the European flat oyster (Ostrea edulis L.).

The European flat oyster (Ostrea edulis L.) represents an economically important oyster production in Southern Italy, widespread in natural beds along the coast. The practice to be eaten raw is an everlasting concern for possible health risk with a need to stringently monitor the health of aquatic environment. A screening survey using histopathological examination was undertaken by harvesting O. edulis from different sites along the Apulian coast of Italy. Tissue samples of the digestive gland, kidney, gonad, and gill were provided for morphologic study in light microscopy (LM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) analysis. The LM observations revealed spherical cytoplasmic inclusions as basophilic prokaryote colonies in 13/250 oysters. The TEM and SEM confirmed the presence of intracytoplasmic inclusions of Rickettsia-like organisms (RLOs), merely in the epithelial cells of the digestive gland tubule tissues in the 13 oysters. Within intracytoplasmic vacuoles, RLOs exhibited a prokaryotic characteristic ultrastructure with transverse binary fission, a DNA zone full of chromatin fibers and a granular periplasmic ribosome zone. O. edulis were found positive for RLOs in wild oysters from Manfredonia, while the other sites were found free of pathological inclusions. Thus, we present the first report of a Rickettsia-like infection in the Apulian wild oyster (O. edulis) from Italy, including an ultrastructural description and pathological characterization.

Thymidine Phosphorylase Expression and Microvascular Density Correlation Analysis in Canine Mammary Tumor: Possible Prognostic Factor in Breast Cancer.

Purpose: The thymidine phosphorylase (TP) is a key enzyme involved in the metabolism of pyrimidines. Inhibition or downregulation of this enzyme causes accumulation of metabolites with consequences in DNA replication. TP regulates angiogenesis and chemotactic activity of endothelial cells. Different studies showed the presence of TP upregulation in human cancer but the correlation between TP expression and the microvascular density (MVD) in canine mammary tumors is unknown. The aim of this study was to investigate a possible correlation between the MVD and TP expression in tumor cells of canine mammary tumors of different degree of severity (G1-G3) by immunohistochemical analysis. Methods: Sixty-eight samples of spontaneous mammary neoplasia of 5-12 cm in diameter were collected from purebred and mixed-breed dogs (mean aged = 9.5 ± 7), not subject to chemotherapy treatments in veterinary clinics. Histopathological analysis and immunostaining were performed. Results: Carcinoma simple samples have been classified as 72.06% of tubule-papillary, 20.59% cysto-papillary, and 7.35% tubular carcinomas. Immunostainings revealed a marked cytoplasmic expression of TP in 30.88% of samples, mild in 32.35%, weaker in 22.07%, and negative in 14.70%. The correlation analysis and two-way ANOVA showed a linear correlation between MVD and TP with a coefficient of correlation (r) > 0.5 (p < 0.05) in G2 and G3. No correlation between variables was found in G1. Conclusions: These findings suggest that cytoplasmic TP overexpression is correlated with microvascular density in canine mammary tumors, in severe grade, and it can be a potential prognostic factor in breast cancer.

Neurological symptoms and mortality associated with Streptococcus gallolyticus subsp. pasteurianus in calves.

Suppurative meningitis-meningoencephalitis (M-ME) is a sporadic disease in neonatal ungulates and only a few studies have reported the involvement of Streptococcus bovis/Streptococcus equinus complex (SBSEC) members in bovine neonatal M-ME. The SBSEC taxonomy was recent revised and previous biotype II/2 was reclassified as S. gallolyticus subsp. pasteurianus (SGP). The aim of this study was to describe a case of fatal neonatal neurological syndrome associated with SGP in calves. Ten calves were monitored because of neurological hyperacute symptoms associate with bilateral hypopyon and death. They were not fed with maternal colostrum; two of them died and were subjected to bacteriological, histopathological and biomolecular analysis as well as antibiotic susceptibility test. Both animals presented lesions mostly concentrated to meninges and brain and had bilateral hypopyon. Nine strains isolated in purity from brain, ocular humors and colon were identified as S. bovis group by using the API Strep system and as S. gallolyticus by using the 16S rRNA sequence. Two of these strains where subjected to WGS analysis that confirmed the sub-species identification and the clonality of the two SGP strains. The strains were found resistant to OT, SXT, MTZ and EN and susceptible to AMP, AMC, KZ and CN. We hypothesized that the syndrome observed could be due to the lack of maternal colostrum feeding. A timely and precise diagnosis could have likely prevented the death of the calves and, since the zoonotic potential of SBSECs members is known, accurate and rapid identification is required.

Sarcocystis bertrami in skeletal muscles of donkeys (Equus africanus asinus) from Southern Italy.

Among the protozoa of the genus Sarcocystis (Apicomplexa; Sarcocystidae), Sarcocystis bertrami (syn. Sarcocystis fayeri) is an obligate intracellular parasite of donkeys and horses with worldwide distribution. Here, we report the detection of S. bertrami in naturally infected donkeys from southern Italy and describe their structure by light microscopy (LM) and transmission electron microscopy (TEM). Protozoal cysts were detected both morphologically and molecularly in skeletal muscles of 28.57% (40/140) donkeys. Mature cysts of S. bertrami were found in skeletal muscle measuring 31-102 µm long and 19-83 µm wide with radially striated thick cyst wall. The high prevalence of infected donkeys suggests that dogs, the definitive hosts of S. bertrami, are contaminating environment with environmentally resistant sporocysts. Considering the increased consumption of raw donkey meat results also suggest a potential risk for human health.