Silver nanoparticles (AgNPs) are an important class of nanomaterials used as antimicrobial agents for a wide range of medical and industrial applications. However toxicity of AgNPs and impact of their physicochemical characteristics in in vivo models still need to be comprehensively characterized. The aim of this study was to investigate the effect of size and coating on tissue distribution and toxicity of AgNPs after intravenous administration in mice, and compare the results with those obtained after silver acetate administration. Male CD-1(ICR) mice were intravenously injected with AgNPs of different sizes (10 nm, 40 nm, 100 nm), citrate-or polyvinylpyrrolidone-coated, at a single dose of 10 mg/kg bw. An equivalent dose of silver ions was administered as silver acetate. Mice were euthanized 24 h after the treatment, and silver quantification by ICP-MS and histopathology were performed on spleen, liver, lungs, kidneys, brain, and blood. For all particle sizes, regardless of their coating, the highest silver concentrations were found in the spleen and liver, followed by lung, kidney, and brain. Silver concentrations were significantly higher in the spleen, lung, kidney, brain, and blood of mice treated with 10 nm AgNPs than those treated with larger particles. Relevant toxic effects (midzonal hepatocellular necrosis, gall bladder hemorrhage) were found in mice treated with 10 nm AgNPs, while in mice treated with 40 nm and 100 nm AgNPs lesions were milder or negligible, respectively. In mice treated with silver acetate, silver concentrations were significantly lower in the spleen and lung, and higher in the kidney than in mice treated with 10 nm AgNPs, and a different target organ of toxicity was identified (kidney). Administration of the smallest (10 nm) nanoparticles resulted in enhanced silver tissue distribution and overt hepatobiliary toxicity compared to larger ones (40 and 100 nm), while coating had no relevant impact. Distinct patterns of tissue distribution and toxicity were observed after silver acetate administration. It is concluded that if AgNPs become systemically available, they behave differently from ionic silver, exerting distinct and size-dependent effects, strictly related to the nanoparticulate form.

Until now, most research has focused on the development of indicators of negative welfare, and relatively few studies provide information on valid, reliable, and feasible indicators addressing positive aspects of animal welfare. However, a lack of suffering does not guarantee that animals are experiencing a positive welfare state. The aim of the present review is to identify promising valid and reliable animal-based indicators for the assessment of positive welfare that might be included in welfare assessment protocols for ruminants, and to discuss them in the light of the five domains model, highlighting possible gaps to be filled by future research. Based on the existing literature in the main databases, each indicator was evaluated in terms of its validity, reliability, and on-farm feasibility. Some valid indicators were identified, but a lot of the validity evidence is based on their absence when a negative situation is present; furthermore, only a few indicators are available in the domains of Nutrition and Health. Reliability has been seldom addressed. On-farm feasibility could be increased by developing specific sampling strategies and/or relying on the use of video- or automatic-recording devices. In conclusion, several indicators are potentially available (e.g., synchronisation of lying and feeding, coat or fleece condition, qualitative behaviour assessment), but further research is required.

This paper reviews animal-based welfare indicators to develop a valid, reliable, and feasible on-farm welfare assessment protocol for dairy goats. The indicators were considered in the light of the 4 accepted principles (good feeding, good housing, good health, appropriate behavior) subdivided into 12 criteria developed by the European Welfare Quality program. We will only examine the practical indicators to be used on-farm, excluding those requiring the use of specific instruments or laboratory analysis and those that are recorded at the slaughterhouse. Body condition score, hair coat condition, and queuing at the feed barrier or at the drinker seem the most promising indicators for the assessment of the "good feeding" principle. As to "good housing," some indicators were considered promising for assessing "comfort around resting" (e.g., resting in contact with a wall) or "thermal comfort" (e.g., panting score for the detection of heat stress and shivering score for the detection of cold stress). Several indicators related to "good health," such as lameness, claw overgrowth, presence of external abscesses, and hair coat condition, were identified. As to the "appropriate behavior" principle, different criteria have been identified: agonistic behavior is largely used as the "expression of social behavior" criterion, but it is often not feasible for on-farm assessment. Latency to first contact and the avoidance distance test can be used as criteria for assessing the quality of the human-animal relationship. Qualitative behavior assessment seems to be a promising indicator for addressing the "positive emotional state" criterion. Promising indicators were identified for most of the considered criteria; however, no valid indicator has been identified for "expression of other behaviors." Interobserver reliability has rarely been assessed and warrants further attention; in contrast, short-term intraobserver reliability is frequently assessed and some studies consider mid- and long-term reliability. The feasibility of most of the reviewed indicators in commercial farms still needs to be carefully evaluated, as several studies were performed under experimental conditions. Our review highlights some aspects of goat welfare that have been widely studied, but some indicators need to be investigated further and drafted before being included in a valid, reliable, and feasible welfare assessment protocol. The indicators selected and examined may be an invaluable starting point for the development of an on-farm welfare assessment protocol for dairy goats.

Outdoor and extensive farming systems allow animals to behave in a natural way and are often perceived as welfare friendly. Nonetheless, the natural environment poses multiple challenges to the welfare of animals, sometimes hampering their capacity to cope. Welfare assessment in outdoor and extensive systems has been rarely investigated, and little is known about the most appropriate indicators. The aim of this review was to identify animal-based measures of welfare to apply in extensive and pasture-based systems in domestic ruminants. Through the use of a dedicated software for systematic reviews, 810 papers were screened and a total of 52 papers were retained for in-depth analysis. ABM resulting from these papers were initially divided according to the species (cattle and small ruminants, including sheep and goats) and then to four principles: comfort, behavior, feeding and health. The results showed that welfare data were collected applying different methodologies, with an increasing use of sensors in recent years. The need to herd and restrain animals for individual data collection is one of the major constraints to data collection in extensive farming systems. It is suggested that welfare assessment in outdoor/extensive farming systems is carried out by following shared procedures in order to provide evidence of the higher animal welfare claims that these products often imply compared to indoor systems.