Virus isolation from saliva and salivary glands of cattle naturally infected with paralytic rabies

The infectivity of saliva, salivary and mammary glands, muscle, lung, kidney and liver of 87 cattle infected with paralytic rabies (positive viral isolation from brains) was studied. Fifty percent dilutions of saliva and tissue samples were inoculated intracerebrally into 10- to 15-day-old mice. Viral isolation in mice was confirmed by direct rabies fluorescent-antibody test and the antigenic variant of the isolates characterized by monoclonal antibodies. Rabies virus was isolated from 4.6% of salivary glands and from 1.6% of saliva samples. The rest of the peripheral tissues were negative. Cerebral and peripheral isolates belonged to vampire-bat antigenic variants. These results indicate that cattle infected by vampire bats may be a source of infection for man. The infection risk would depend on the type of contact between rabid cattle and man.     

Identification and association analysis of several hundred single nucleotide polymorphisms within candidate genes for back fat thickness in Italian Large White pigs using a selective genotyping approach

Combining different approaches (resequencing of portions of 54 obesity candidate genes, literature mining for pig markers associated with fat deposition or related traits in 77 genes, and in silico mining of porcine expressed sequence tags and other sequences available in databases), we identified and analyzed 736 SNP within candidate genes to identify markers associated with back fat thickness (BFT) in Italian Large White sows. Animals were chosen using a selective genotyping approach according to their EBV for BFT (276 with most negative and 279 with most positive EBV) within a population of ≈ 12,000 pigs. Association analysis between the SNP and BFT has been carried out using the MAX test proposed for case-control studies. The designed assays were successful for 656 SNP: 370 were excluded (low call rate or minor allele frequency <5%), whereas the remaining 286 in 212 genes were taken for subsequent analyses, among which 64 showed a P(nominal) value <0.1. To deal with the multiple testing problem in a candidate gene approach, we applied the proportion of false positives (PFP) method. Thirty-eight SNP were significant (P(PFP) < 0.20). The most significant SNP was the IGF2 intron3-g.3072G>A polymorphism (P(nominal) < 1.0E-50). The second most significant SNP was the MC4R c.1426A>G polymorphism (P(nominal) = 8.0E-05). The third top SNP (P(nominal) = 6.2E-04) was the intronic TBC1D1 g.219G>A polymorphic site, in agreement with our previous results obtained in an independent study. The list of significant markers also included SNP in additional genes (ABHD16A, ABHD5, ACP2, ALMS1, APOA2, ATP1A2, CALR, COL14A1, CTSF, DARS, DECR1, ENPP1, ESR1, GH1, GHRL, GNMT, IKBKB, JAK3, MTTP, NFKBIA, NT5E, PLAT, PPARG, PPP2R5D, PRLR, RRAGD, RFC2, SDHD, SERPINF1, UBE2H, VCAM1, and WAT). Functional relationships between genes were obtained using the Ingenuity Pathway Analysis (IPA) Knowledge Base. The top scoring pathway included 19 genes with a P(nominal) < 0.1, 2 of which (IKBKB and NFKBIA) are involved in the hypothalamic IKKβ/NFκB program that could represent a key axis to affect fat deposition traits in pigs. These results represent a starting point to plan marker-assisted selection in Italian Large White nuclei for BFT. Because of similarities between humans and pigs, this study might also provide useful clues to investigate genetic factors affecting human obesity.     

Heritability of muscular cathepsin B activity in Italian Large White pigs

Previous studies suggested that proteolysis lasting for many months after slaughtering could be responsible for excessive softness in dry-cured hams. Also, the long-term activity of muscle proteinases is mainly attributed to cathepsin B. Should an additive genetic component exist for the activity level of cathepsin B, such information could be used as a selection criterion for improving meat quality for dry-cured ham production. Data on the growth rate, carcass composition, and meat quality of 234 Large White pigs raised in a test station under a &lsquo| quasi ad libitum ' nutritive level and slaughtered at around 160 kg live weight were analysed using a blup - mt -animal model. The cathepsin B activity level showed a moderate heritability (0.23–0.28) in all analyses, and negative genetic correlations with daily gain (− 0.34 to − 0.52) and backfat thickness (− 0.40 to − 0.86), but, with large standard errors. These results suggest that a genetic component exists in explaining the variability of the cathepsin B activity level. Further studies are necessary to confirm the outcome and to investigate genetic correlations with other traits currently under selection.     

Metabolic profiling of strawberry grape ( Vitis × labruscana cv. 'Isabella') components by nuclear magnetic resonance (NMR) and evaluation of their antioxidant and antiproliferative properties

In the assessment of the antioxidant properties of edible plants, the widely consumed Vitis × labruscana cv. 'Isabella', known in Italy as "fragola" (strawberry) grape, was of interest. Phenol and flavonoid contents of the methanolic extracts of peel, pulp, seed, leaf, and stalk components of the plant were determined. The metabolic profile of the extracts was performed by 1D and 2D NMR. Quantitative analysis, obtained in the presence of 0.01% of internal standard trimethylsilyl propionate, evidenced the presence of catechins in both stalk and seed extracts, whereas caffeic acid and quercetin were the main metabolites of the leaf extract. Furthermore, the extracts were tested for their radical scavenging and reducing capacities by measuring their capacity to scavenge DPPH(?) and ABTS(?+) and to reduce Fe(III) and Mo(VI) salts. The antioxidant efficacy of the extracts in cell-free systems and their antiproliferative activity toward HepG2 and A549 cells were also evaluated. Seed and stalk components are able to reduce by 39.6 and 40.6%, respectively, the amount of the metabolically active HepG2 cells after only 24 h of exposure.     

Levels of benzo[a]pyrene (BaP) in "mozzarella di bufala campana" cheese smoked according to different procedures

The content of benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon, was determined by HPLC-FL in "mozzarella di bufala campana" cheese, a stretched cooked cheese, either experimentally smoked according to traditional procedures, using straw, cardboard, and wood shavings or aromatized with smoke flavoring. The BaP residues, researched also in cheese samples sold at retail, were detected in the rind, in the core, and in the slice (outer and inner parts). In the cheeses experimentally smoked with straw and cardboard the BaP levels, ranging from 0.38 to 2.12 microg kg(-1) and from 0.46 to 2.40 microg kg(-1), respectively, were statistically higher than those of the cheeses smoked with wood shavings and aromatized with liquid smoke (from 0.19 to 0.80 microg kg(-1) and from 0.18 to 0.50 microg kg(-1), respectively). However the cheeses treated with liquid smoke flavor showed a BaP content exceeding the level allowed by the European Union. In the samples sold at retail, smoked with straw, values were lower than those obtained from samples smoked experimentally with the same combustible. This is probably due to different smoking technologies among the several provinces of the Protected Designation of Origin (PDO) area. PDO is a term used to characterize foodstuffs produced and prepared in a given geographical region by the means of a recognized process. A standardization of the traditional smoking procedures and an improvement of liquid smoke purification treatments are recommended for mozzarella cheese.     

A Nutraceutical Formulation Containing Brown Algae Reduces Hepatic Lipid Accumulation by Modulating Lipid Metabolism and Inflammation in Experimental Models of NAFLD and NASH

Recently, some preclinical and clinical studies have demonstrated the ability of brown seaweeds in reducing the risk factors for metabolic syndrome. Here, we analyzed the beneficial effect of a nutraceutical formulation containing a phytocomplex extracted from seaweeds and chromium picolinate in animal models of liver steatosis of differing severities (rats with non-alcoholic fatty liver disease (NAFLD) and its complication, non-alcoholic steatohepatitis (NASH)). This treatment led to a significant drop in hepatic fat deposition in both models (p < 0.01 vs. untreated animals), accompanied by a reduction in plasma inflammatory cytokines, such as interleukin 6, tumor necrosis factor α, and C reactive protein, and myeloperoxidase expression in liver tissue. Furthermore, a modulation of the molecular pathways involved in lipid metabolism and storage was demonstrated, since we observed the significant reduction of the mRNA levels of fatty acid synthase, diacylglycerol acyltransferases, the sterol-binding protein SREBP-1, and the lipid transporter perilipin-2, in both treated NAFLD and NASH rats in comparison to untreated ones. In conclusion, this nutraceutical product was effective in reducing liver steatosis and showed further beneficial effects on hepatic inflammation and glycemic control, which were particularly evident in rats characterized by a more severe condition, thus representing a therapeutic option for the treatment of NAFLD and NASH patients.     

Detection and phylogenetic characterization of astroviruses in insectivorous bats from Central‐Southern Italy

In recent years, bats have been found to harbour many viruses, raising several questions about their role as reservoirs and potential disseminators of zoonotic viruses. We investigated the presence of six virus families in bats in three regions of Central‐Southern Italy. Astroviruses were identified in seven of 13 bat species. Sequence analysis revealed marked genetic heterogeneity among the astroviruses identified, with nucleotide identity ranging between 60.26% and 87.62%. Astrovirus diversity was not associated with the bat species, the geographic areas or the bat colony, suggesting the circulation of several astrovirus strains in Italian ecosystems. Genetic diversification and interspecies transmission appear common in bat astroviruses and could provide, potentially, the bases for transmission to humans and other mammals. Yet overemphasizing this risk might have detrimental consequences for bat conservation and preservation of the important ecosystem services bats provide.