Short communication: Seroprevalence of paratuberculosis in Italian water buffaloes (Bubalus bubalis) in the region of Campania.

Paratuberculosis is a chronic enteric disease affecting virtually all ruminants, but only anecdotal information is currently available about the occurrence of this disease in water buffaloes (Bubalus bubalis). We carried out a survey study aimed at determining the prevalence of paratuberculosis in 2 provinces in the region of Campania, Italy, where about half of all Italian buffaloes are reared. From May 2017 to December 2018, we collected 201,175 individual serum samples from 995 buffalo herds. The sera were collected from animals over 24 mo old and were tested using a commercial ELISA test. The herd-level apparent prevalence result was 54.7%, and the animal-level apparent prevalence was 1.8%. The herd-level true prevalence was estimated using a Bayesian approach, demonstrating a high herd-level prevalence of paratuberculosis in water buffaloes from the Campania area. These findings suggest that the urgent adoption of paratuberculosis herd-control programs for water buffaloes in this area would be beneficial.

Short communication: comparing real-time PCR and bacteriological cultures for Streptococcus agalactiae and Staphylococcus aureus in bulk-tank milk samples.

For more than 30 yr, a control plan for Streptococcus agalactiae and Staphylococcus aureus has been carried out in more than 1,500 dairy herds of the province of Brescia (northern Italy). From 2010 to 2011, the apparent prevalence of Strep. agalactiae has been relatively stable around 10%, but the apparent prevalence of Staph. aureus has been greater than 40% with an increasing trend. The aim of this paper was to estimate and compare the diagnostic accuracy of 3 assays for the detection of Strep. agalactiae and Staph. aureus in bulk-tank milk samples (BTMS) in field conditions. The assays were a qualitative and a quantitative bacteriological culture (BC) for each pathogen and a homemade multiplex real-time PCR (rt-PCR). Because a gold standard was not available, the sensitivities (Se) and specificities (Sp) were evaluated using a Bayesian latent class approach. In 2012 we collected one BTMS from 165 dairy herds that were found positive for Strep. agalactiae in the previous 2-yr campaigns of eradication plan. In most cases, BTMS collected in these herds were positive for Staph. aureus as well, confirming the wide spread of this pathogen. At the same time we also collected composite milk samples from all the 8,624 lactating cows to evaluate the within-herd prevalence of Strep. agalactiae. Streptococcus agalactiae samples were cultured using a selective medium Tallium Kristalviolette Tossin, whereas for Staph. aureus, we used Baird Parker modified medium with added Rabbit Plasma Fibrinogen ISO-Formulation. In parallel, BTMS were tested using the rt-PCR. Regarding Strep. agalactiae, the posterior median of Se and Sp of the 2 BC was similar [qualitative BC: Se=98%, posterior credible interval (95%PCI): 94-100%, and Sp=99%, 95%PCI: 96-100%; quantitative BC: Se=99%, 95%PCI: 96-100%, and Sp=99%, 95%PCI: 95-100%] and higher than those of the rt-PCR (at 40 cycle threshold, Se=92%, 95%PCI: 85-97%; Sp=94%, 95%PCI: 88-98%). Also in case of Staph. aureus, the posterior medians of BC were generally higher than those of rt-PCR. In fact, although the Se of BC was slightly lower (rt-PCR at 40 cycle threshold, median Se=99%, 95%PCI: 97-100%, and qualitative BC, median Se=94%, 95%PCI: 87-99%), the Sp was much higher (rt-PCR at 40 cycle threshold, median Sp=67%, 95%PCI: 38-97%; qualitative BC, median Sp=95%; 95%PCI: 76-100%). Our study confirms that BC and rt-PCR are reliable diagnostic tools to detect Strep. agalactiae and Staph. aureus, and rt-PCR results should be confirmed by BC carried out on BTMS and possibly on composite milk samples.

A method to identify the areas at risk for the introduction of avian influenza virus into poultry flocks through direct contact with wild ducks.

Wild dabbling ducks are the main reservoir for avian influenza (AI) viruses and pose an ongoing threat to commercial poultry flocks. Combining the (i) size of that population, (ii) their flight distances and (iii) their AI prevalence, the density of AI-infected dabbling ducks (DID) was calculated as a risk factor for the introduction of AI viruses into poultry holdings of Emilia-Romagna region, Northern Italy. Data on 747 poultry holdings and on 39 AI primary outbreaks notified in Emilia-Romagna between 2000 and 2017 were used to validate that risk factor. A multivariable Bayesian logistic regression was performed to assess whether DID could be associated with the occurrence of AI primary outbreaks. DID value, being an outdoor flock, hobby poultry trading, species reared, length of cycle and flock size were used as explanatory variables. Being an outdoor poultry flock was significantly associated with a higher risk of AI outbreak occurrence. The probability of DID to be a risk factor for AI virus introduction was estimated to be 90%. A DID cut-off of 0.23 was identified to define high-risk areas for AI virus introduction. Using this value, the high-risk area covers 43% of the region. Seventy-four per cent of the primary AI outbreaks have occurred in that area, containing 39% of the regional poultry holdings. Poultry holdings located in areas with a high DID value should be included in a risk-based surveillance programme aimed at AI early detection.

B cells and macrophages pursue a common path toward the development and progression of chronic lymphocytic leukemia.

The development and progression of chronic B-cell tumors depend on a complex microenvironmental network of cells that include monocyte-derived macrophages. In chronic lymphocytic leukemia (CLL) the survival of malignant cells is supported in vitro by nurse-like cells, which differentiate from CD14+ monocytes and have been identified as tumor-associated macrophages (TAMs). The role of the monocyte/macrophage lineage in CLL has been extensively studied in vitro, but only recently has been investigated in in vivo models. We here discuss how the cellular and molecular interactions that physiologically occur between B cells and macrophages can be subverted in chronic B lymphoid malignancies. Clinical approaches for the therapeutic targeting of TAMs are under evaluation. Promising strategies, along with a direct impact on the malignant cells, affect crucial pathways involved in the interaction of leukemic cells with TAMs. As an example, ibrutinib reduces CLL cell chemoattraction by inhibiting macrophage secretion of CXCL13. Lenalidomide and trabectedin prevent TAM recruitment mainly through CCL2 blockade. Most advanced strategies aim at depleting macrophages by targeting the CSF1/CSF1R pathway, which is fundamental for TAM survival. Of note, CSF1 transcripts are significantly more abundant in progressive CLL patients when compared with stable CLL and the frequency of CSF1R+ TAMs correlates with poor survival in hematological malignancies. The successful combination of CSF1R inhibition with currently available agents targeting malignant cells might represent the next therapeutic frontier in CLL. Conceivably these approaches may become applicable to numerous chronic B lymphoid malignancies.

Low-pass filtering, a new method of fractal analysis: application to PET images of pulmonary blood flow.

The pattern of a spatial structure that repeats itself independently of the scale of magnification or resolution is often characterized by a fractal dimension (D). Two-dimensional low-pass filtering, which may serve as a method to assess D, was applied to functional images of pulmonary perfusion measured by positron emission tomography. The corner frequency of a low-pass filter is inversely proportional to the resolution scale. The method was applied to three types of images: random noise images, synthetic fractal images, and positron emission tomographic images of pulmonary perfusion. Images were processed with two-dimensional low-pass filters of decreasing corner frequencies, and a spatial heterogeneity index, the coefficient of variation, was calculated for each low-pass-filtered image. The natural logarithm of the coefficient of variation scaled linearly with the natural logarithm of the resolution scale for the PET images studied (average R(2) = 0.99). D ranged from 1.25 to 1.36 for the residual distribution of pulmonary perfusion after vertical gradients were removed by linear regression. D of the same data without removal of vertical gradients ranged from 1.11 to 1.14, but the fractal plots had systematic deviations from linearity and a lower linear correlation coefficient (R(2) = 0. 96). The method includes all data in the lung field and is insensitive to the effects of misregistration. We conclude that low-pass filtering offers new insights into the interpretation of D of two-dimensional functional images as a measure of the frequency content of spatial heterogeneity.

High performance liquid chromatography versus gas chromatography in the analysis of proteinaceous painting ligands.

High performance liquid chromatography (HPLC) and gas chromatography (GC) were compared for the analysis of the amino acid composition in painting ligands.

The effects of heparin and dietary fish oil on embolic events and the microcirculation downstream from a small-artery repair.

In recent studies we demonstrated that platelet emboli induced by arterial injury impair the microcirculation. The present study was performed to determine if heparin or dietary cod liver oil reduces the incidence of emboli following the same arterial injury and, in turn, whether these agents prevent the associated decrease in microcirculatory blood flow. The cremaster muscle of 29 male Sprague-Dawley rats was isolated on a single neurovascular bundle consisting of the iliac artery and vein and the genitofemoral nerve. Emboli were generated by a thrombogenic injury of the iliac artery, and their number and their subsequent effect on capillary perfusion in the downstream microcirculation were measured. Nine animals received heparin (10-unit IV bolus plus 10 units/hour IV infusion), 10 were fed cod liver oil (10 percent by weight of food) for 3 weeks prior to the experiment, and 9 animals receiving no treatment served as controls. The number of emboli was significantly reduced in the heparin group, but there was no accompanying improvement in capillary perfusion. In contrast, in the cod liver oil group, the number of emboli was not reduced, but there was significant improvement in capillary perfusion. These findings suggest that the harmful effect that platelet emboli have on the microcirculation is probably biochemical in nature (vasoconstriction) rather than related to simple mechanical obstruction to flow.

Fibrin sealant coated stents compared with non-coated stents in a porcine carotid artery model. Preliminary study report.

BACKGROUND: Balloon expandable metal stents (BEMS) are used to treat restenosis following percutaneous transluminal angioplasty (PTA) and as primary treatment. Intimal proliferation (IP) and resultant restenosis occurs in 25-50% of patients despite all preventive measures. OBJECTIVE: to test the intra-arterial response to the insertion of a fibrin sealant (FS) coated BEMS vs an uncoated BEMS by measuring endothelization and IP. HYPOTHESIS: that a BEMS coated with FS will lead to rapid endothelization and prevent or reduce IP. RATIONALE: FS consists of fibrinogen and thrombin. Thrombin affects endothelial cell proliferation and reduces smooth muscle proliferation, the forerunner of IP and restenosis. Normal endothelium also releases substances that promote vascular relaxation and normal smooth muscle tone regulation. METHODS: Thirty-40 kg pigs (EA), Palmaz-Shatz BEMS (Cordis), FS Tissucol (Baxter Immuno). Stents were uniformly coated with FS in a special mold. Both coated and uncoated stents were mounted on balloon catheters and deployed caudad in the carotid arteries via an arteriotomy. Angiograms were obtained postdeployment. All specimens were examined grossly, photographed then fixed for histology and in some cases, scanning electron microscopy (SEM). RESULTS: Fifteen animals form this preliminary report. Sacrifice at five days as per original protocol showed insufficient stent incorporation. Thereafter 1/2 of the animals were sacrificed at 15 days and 1/2 at 30 days. PATENCY: coated stents: 6 patent, 9 thrombosed. UNCOATED: 7 patent, 8 thrombosed. Of five EA given postoperatively low molecular weight heparin (LMWH) 4 animals had patent stents 80%. HISTOLOGY: varying degrees of IP were seen in all specimens. In general the coated stents showed a greater degree than the uncoated. STENOSIS: presacrifice angiography revealed that where the stents were patent no stenosis was present, in fact, some demonstrated mild dilatation. This was particularly the case with the coated stents. CONCLUSIONS: Coating stents with FS is not detrimental. IP in these EA at 30 days did not produce stenosis. Postoperative LMWH appears helpful in maintaining patency in a thrombogenic experimental animal. Further study maintaining EA for 6-12 months should resolve whether the IP seen had achieved its maximum expression or would progress and produce stenosis.

Determination of polycyclic aromatic hydrocarbons in sediment using solid-phase microextraction with gas chromatography-mass spectrometry.

Manual solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS) is applied for the determination of polycyclic aromatic hydrocarbons (PAHs) from natural matrix through a distilled water medium. Seven of the 16 PAH standards (naphthalene, acenaphthene, fluorene, anthracene, fluoranthene, pyrene, benzo[a]anthracene) are spiked on a marine muddy sediment. The samples, containing PAHs in the range of 10-20 ppm, are then aged at room temperature more than 10 days before analysis. The influence of the matrix, SPME adsorption time, pH, salt content, and SPME adsorption temperature are investigated. The reproducibility of the technique is less than 13% (RDS) for the first 6 considered PAHs and 28% (RDS) for benzo(a)anthracene with a fiber containing a 100-micron poly dimethylsiloxane coating. Linearity extended in the range of 5-50 picograms for PAHs direct injection, 5-70 picograms for PAHs in water, and 1-170 picograms for PAHs in sediment. The detection limit is estimated less than 1 microgram/kg of dry sample for the first 6 considered PAHs in sediment and 1.5 micrograms/kg of dry sample for benzo(a)anthracene using the selected ion monitoring mode in GC-MS. The recoveries of the considered PAHs are evaluated.

[Toxic occupational liver diseases. Therapeutic effects of silymarin]. / Epatopatie tossiche professionali. Effetti terapeutici della silimarina.

We considered two groups, one of 35 and the other of 20 patients, with occupational toxic hepatopathy caused by various toxic substances (mostly solvents, paints and glues). The patients were mostly suffering from chronic or subacute forms. We considered the work conditions in the anamnesis, the period of exposure and most important laboratory parameters of hepatic function checked before and after treatment with Silymarin (420 mg/die/os) for the first group and with "placebo" for the second group. Five patients of the first group were diagnosed by biopsy. The treatment with Silymarin has shown slight variations in some parameters. The therapeutic effect is probably not dependent upon the kind of pathogen noxa; it seems instead to be more evident when the exposure period is shorter. The group "placebo" does not show significant variations.

[Surgical treatment of hydatid cysts]. / Trattamento chirurgico delle cisti idatidee complicate.

Hydatidosis represents a form of helminthic zoonosis with a stronger medical, social and economic impact. It is endemic in some areas of the world including Mediterranean countries, and therefore Italy. The disease is often diagnosed in an advanced phase, due to the poor symptomatology in the early stages. The best treatment is surgery. Emergency surgery is similar to elective surgery, though in some cases is preferable only a drainage procedure until the stabilization of the genreal and local conditions of the patient allow a radical intervention. The Authors report on their five year experience of surgical treatment of compliated pulmonar hydatidosis pointing out that exeresis must be aimed at preserving pulmonar function as much as possible.

[Combined video-thoracoscopic, surgical and endovascular treatment in a case of ascites and recurrent bilateral pleural effusion]. / Trattamento combinato video-toracoscopico, chirurgico ed endovascolare in un caso con ascite e versamento pleurico bilaterale recidivante.

Among recurrent pleural effusions a role of remarkable importance is held by those combined with ascitis due to the difficulty of their treatment, even using widely tested techniques. The incidence of such pathology varies from 4% to 6% of patients suffering from cirrhotic pathology, reaching 10% in cases with advanced illnesses. Pleural effusions involve the right emithorax more frequently than the left one, but it can show up bilaterally too. Its etiopathogenesis is tied up to the direct passage of ascitic liquid into the chest and, during the past years, numerous theories have been described to explain this migration. The Authors report the case of a patient with interesting considerations for the diagnostic difficulties and the peculiarity of the treatment performed.

[Surgical treatment of celomic cysts]. / Trattamento chirurgico delle cisti celomatiche.

Celomic cysts (pericardiopleural or serous cysts) are rare, as they account for about 30% of all the homoplastic disembryogenetic lesions of the mediastinum. There is not usually a characteristic symptomatology associated with this anomaly, which is, however, easily diagnosed by means of the most common radiologic techniques. The TC together with the MRI provides all the necessary data for an appropriate management of the patient. The Authors report on their five year experience of the surgical treatment of celomic cysts, pointing out that surgery must be aimed at preserving pulmonary function as much as possible. They finally maintain that the first surgical approach should be video-thoracoscopic with the aid of a minithoracotomy, if necessary.