Pier Paolo Danieli

Aflatoxin B1 and fumonisin B1 affect the oxidative status of bovine peripheral blood mononuclear cells

Mycotoxins are secondary metabolites having a high cytotoxic potential. They are produced by molds and released in food and feed. To date, the mechanisms underlying the mycotoxin-induced cytotoxicity have not been fully clarified. The induction of oxidative stress, as a possible mechanism, has been postulated. This in vitro study was focused on the effect of two widely occurring mycotoxins, aflatoxin B(1) (AFB(1)) and fumonisin B(1) (FB(1)), on the oxidative status of bovine peripheral blood mononuclear cells (PBMC) incubated for 2 and 7 days at different levels of AFB(1) (0, 5 and 20 μg/ml) and FB(1) (0, 35 and 70 μg/ml). Reactive oxygen metabolites (ROM), intracellular thiols (SH), malondialdehyde (MDA) and gene expression of cytoplasmic superoxide dismutase (SOD) and glutathione peroxidase (GSHPX-1) were measured on PBMC after incubation. The highest concentration of AFB(1) and all concentrations of FB(1) caused an increase (p<0.05) of intracellular ROM without any time dependent effect. Intracellular SH decreased with 20 μg AFB(1)/ml (p<0.05) and the effect was particularly marked after 7 days of exposure. Intracellular SH were not affected by FB(1) even though a lower (p<0.05) SH level after 2 days exposure than after 7 days was observed. MDA increased (p<0.05) in AFB(1) or FB(1) treated PBMC. The exposure to FB(1) for 7 days increased MDA (p<0.05) only in cells treated with 70 μg/ml. Exposure of PBMC to AFB(1) reduced SOD mRNA while FB(1) decreased both SOD and GSHPX-1 mRNA abundance. These results demonstrate that, even though by different mechanisms, AFB(1) and FB(1) may induce cytotoxicity through an impairment of the oxidative status of PBMC.

Quality evaluation of regional forage resources by means of near infrared reflectance spectroscopy

Abstract Quality parameters of grassland and pasture samples collected during a three-year period at two environmentally and geographically different areas were analysed by Near Infrared Reflectance Spectroscopy (NIRS). Chemical analysis for crude protein (CP), crude fibre (CF), neutral detergent fibre (NDF), acid detergent fibre (ADF), acid detergent lignin (ADL) and crude ash (ASH) carried out on two-thirds of the samples were used in calibration processes. The remaining one-third of the data was used to validate the best calibrations obtained. Samples selection is discussed. Different math pre-treatments (derivative, gap, primary smoothing and secondary smoothing), light scattering correction methods and calibration algorithms were tested to achieve the better predictive performances. We obtained the best results using different regression algorithms to correlate spectral information to chemical data. For CP (R2 = 0.94, SEP=1.3), NDF (R2 = 0.95, SEP = 2.14) and ADF (R2 = 0.92, SEP=2.06) Multiple Linear Regression (MLR) models fit chemical data better than Mean Partial Least Square (MPLS) regression. A molecular basis explanation of wavelengths selected was carried out. MPLS models worked well for CF (R2 = 0.93, SEP=1.57), and ASH (R2 = 0.95, SEP=1.17) while poor calibrations were obtained for ADL using both algorithms. To confirm the reliability of the models developed, uncertainties of predictions were compared with findings on nutritional variations and animal performances.

An update on the safety of foods of animal origin and feeds

Chemical hazards may occur in any phases of the different livestock production systems. Aim of this review is to address an update about the key issues related to the risk of contamination in foods of animal origin by environmental contaminants linked to industrialisation or urbanisation (e.g., heavy metals and persistent organic pollutants), and natural contaminants (e.g., mycotoxins). This review deals with current issues and future perspectives on the complex issue of the safety of feeds and foods of animal origin, by taking into account the estimation of the occurrence of chemical residues in food, the hazard identification and characterisation of mycotoxins in animal feeds, and the analysis of feedstuffs as a tool to control and evaluate food safety.

Benthic diatom assemblages and their response to human stress in small-sized volcanic-siliceous streams of central Italy (Mediterranean eco-region)

The river typology ‘volcanic rivers’ represents a unique typology in the Mediterranean eco-region in Europe but its type-specific diatom assemblages are still poorly investigated. The main aims of this study were (i) to analyse the diatom diversity of this river type, (ii) to investigate the relationships between diatom diversity and environmental variables and (iii) to identify the characterizing diatom community structure of different environmental groups. To this end, we selected 13 streams located in central Italy, belonging to a single river type and covering the whole quality range from human impacted sites to those in ‘reference conditions’. Most frequent and abundant diatom species found were Cocconeis placentula var. lineata, Planothidium lanceolatum, Rhoicosphenia abbreviata and Amphora pediculus. Species richness in reference and less impacted sites was not higher than in intermediate and heavily degraded ones, but non-metric multidimensional scaling (N-MDS) analysis revealed a substantial difference among their diatom community structure. The indicator species analysis defined the characteristic species of three environmental groups identified by N-MDS. The findings of this study represent a contribution to the knowledge on diatom communities of Mediterranean volcanic-siliceous rivers and provide information for a data-driven implementation of the Water Framework Directive in Italy for this specific river typology.

Contamination by Persistent Chemical Pesticides in Livestock Production Systems

The impact of persistent organic pesticides represents one of the major environmental problems as reported in several studies and reflected in some mandatory actions at the inter-governmental level. In particular, isomers of Hexachlorocyclohexane (HCH), like many others Organochlorine Pesticides (OCPs), are of human health and environmental concern due to their persistence in the biosphere. In industrialised countries, Lindane (the γ-isomer of HCH with insecticide effects) has been widely used in the past. As a consequence, large quantities of HCH isomers without insecticide effects (α-, β-, δ- and e), discarded during the purification of Lindane, have been disposed for years into dumps around industrial sites. Nowadays, such uncontrolled disposal practices imply a great risk of environmental contamination, possibly threatening animals and humans by food chains transfer and bio-accumulation. In 2005, dairy cow farms in the Province of Rome (Italy) were threatened due to environmental contamination of the Sacco River by HCHs. As a case study, all components of the dairy cow production system undergoing agroenvironmental pollution crisis, were investigated with the aim to analyze the main critical points. Five dairy farms were involved in the research. Data regarding the contamination by HCHs of soils, forages, bovine milk and blood serum are reported. Soil and forage samples (mainly maize, alfalfa and ryegrass) were taken in different places near the Sacco River, on the basis on irrigation practiced and flooding conditions. All samples were analyzed by Gas Cromatography using an Electron Capture Detector. Soil contamination by HCHs was found higher nearby than away the river (p < 0.01) with a great incidence of outflow risk (p < 0.01), while no differences were observed on the basis of irrigation practices. In alfalfa samples higher concentration of HCHs than in ryegrass were detected, with a greater plant/soil apparent partition factor. Differences in milk contamination by β-HCH among dairy farms (p < 0.01) and sampling time were found (p < 0.05). In many cases, the β-HCH content of milk resulted above the EU limit (0.003 mgβ-HCH/kg), posing serious hazard for human consumption due to chronic toxicity of that isomer. Differences in milk β-HCH concentration were found related to lactating phase and parity. A linear regression between blood serum and milk β-HCH concentration was observed (r2 = 0.919, p < 0.05). Furthermore, β-HCH as a trace was detected in blood sera when milk levels fell below the analytical limits, indicating the usefulness of blood serum HCH content as an early indicator of animal exposure. Results obtained from the case study highlighted the needing for further researches at wider level, due to the strong impact of persistent organic pesticides on rural environment and human food chain.

Evaluation of mycotoxins contamination in intensive beef cattle production system

Abstract Feed samples from 15 intensive beef cattle farms located in Northern Italy (provinces of Verona and Mantova) were checked for mycotoxin contamination [aflatoxins (AF), fumonisins (FB), ochratoxin A (OTA), deoxynivalenol (DON) and zearalenol (ZEN)]. Total mixed rations (TMR) resulted positive for AF and FB contamination. Among feedstuffs corn and corn gluten feed were the only responsible of TMR contamination. Level of contamination was positively related to corn moisture content.

Stable isotopes in tissues discriminate the diet of free-living wild boar from different areas of central Italy.

The use of isotopic signatures in animal tissues provides information on the environment where they are living and, notably, on their diet. Carbon and, whenever possible, nitrogen stable isotope analyses were performed in animal hairs, muscles and fat. Particularly, we analyzed both carbon and nitrogen isotopic compositions (δ13C and δ15N) on wild boar samples across three different areas of central Italy (Latium region): Tyrrhenian Coast (TC), Maremma (MA) and Central Plains (CP). The agricultural habits of these areas imply that, in winter, no crops are available for wild boars, which feed mainly on acorns and natural feeds (tubers, earthworms etc.). In addition, the three areas were influenced by oak masting. One of these areas (CP) was characterised by the spreading of corn during the hunting season to attract the animals. For each area, we sampled 10 animals aged between 12 and 24 months and balanced by gender. Anenrichment of δ13C in CP area, where corn was used, was observed in all the analysed tissues in comparison to other areas (MA and TC). In CP area, enriched values of δ15N were also observed in all the tissues. The research demonstrates that both δ13C andδ15N in free-living wild boar tissues are influenced by sampling area. According to feeding habits of the species and wildlife management (feed supplementation), the differences observed in δ13C and δ15Nare based on the specific feeding regime; particularly the use of corn in wintertime. Furthermore, the research highlights and discusses diversities and relationships among δ13C and δ15N in the hair, fat and muscles of free-living wild boar.

Conventional and homeopathic treatments in late pregnant goats: effects on metabolic status and immune response

Abstract The study was aimed at assessing the effects of conventional and homeopathic treatments on metabolic status and immune response in late pregnant goats. Administration of an antichetogenic preparation and of Echinacea purpurea in homeopathic dilution did not exert unequivocal effects on metabolic status, but improved some immunological parameters of periparturient goats.

Influence of dietary vitamin E supplementation on cholesterol oxidation and fresh colour in beef aged for 3 and 14 days

Abstract The effects of dietary vitamin E supplementation on the susceptibility to lipid oxidation and colour of the Longissimus thoracis (LT) muscle aged in vacuum packaged conditions for 3 or 14 days were studied. For this purpose, Charolais cattle were fed on a diet providing daily 60 mg (control) or 5500 mg of vitamin E per animal (supplemented) for 30 and 60 days before slaughter. Dietary vitamin E supplementation increased the liver vitamin E content, but not in the LT muscle of treated animals. The vitamin supplementation for 30 and 60 days has shown non-consistent effects in reducing cholesterol oxidation products of vacuum-packed aged meat. However, the vitamin E supplementation for 60 days was effective on Lightness stability in LT muscle during vacuum-packed ageing. Overall, from the practical standpoint, this study suggests that supra-nutritional supplementation up to 60 days may not increase the vitamin E content of Charolais LT muscle giving little, if any, benefits on meat colour and cholesterol oxidation. However, the present study suggests that it would be interesting to determine in which extent specific oxy-sterols are related to the meat colour and whether colour parameters can be useful for predicting the formation of cholesterol oxidation products along the industrial meat production chain.

In vitro effects of the organochlorine pesticide β-hexachlorocyclohexane on bovine peripheral blood mononuclear cells

The β-hexachlorocyclohexane (β-HCH) is a very stable and accumulable isomer of Lindane, a well known organochlorine pesticide. The HCHs were banned in all developed countries but to date high concern still exists for environment, animal and human health due to contaminated sites. In this study, several in vitro tests [cell viability (XTT), trypan blue exclusion (TBE), lactate dehydrogenase release (LDH) and bromodeoxyuridine (BrdU) incorporation assays] were performed to investigate the toxic effects of exposure to β- HCH (from 0.1 to 1000 μM) on bovine peripheral blood mononuclear cells (PBMCs). All the trials were performed incubating PBMCs for 2 and 7 days. At high concentrations (i.e. 1000 μM), the β-HCH approximately halved the number of living cells regardless the exposure time, significantly decreased the cell viability assessed by the XTT assay, and compromised the proliferation potential of PBMCs. At lower β-HCH exposure levels (0.1 to 100 μM), particularly after 7 days of exposure, a progressive decrease of cell viability has been observed. These adverse effects were significant at concentrations observed in the blood of cattle reared in polluted areas. The LDH results suggest that β-HCH does not clearly affect the integrity of the cell membrane in the range of exposure levels tested. All in all, these findings warn about the risk posed by the long-term exposure to β-HCH of farm animals reared in rural areas polluted by β-HCH. Further research is needed to deepen our knowledge about the mechanisms through which β-HCH affects the PBMCs functionality.