A methodological approach to compare continuous and instantaneous sampling and two methods to deal with animals out of sight on dairy cattle behavior and interaction with their calf in the first hours post-partum.

Animal behavior can provide useful information about animal welfare, but methods and tools used to gather behavioral data and data treatment can influence the results. Therefore, this study was carried out on dairy cow (Bos taurus) behavior and interaction with calves early post-partum aiming at comparing two sampling rules, namely continuous and instantaneous sampling at scan intervals of 30 s, 1, 2, 3, 4, 5, and 10 min, and of two methods to deal with out of sight animals. The study was based on three assumptions: (1) continuous sampling provides the most complete and accurate data, allowing the observation of seldom behaviors and short events; (2) instantaneous sampling can provide accurate measurements of frequency and duration, especially at short scan intervals; (3) differences in behavioral results may occur depending on whether a correction for time out of sight is applied or not. Thus, 10 dams were observed from videos in the 2 h post-parturition. Ruminating, stereotypies, calf-biting and calf-butting were not recorded during the observation period. Other behaviors were observed only with continuous sampling or with continuous and instantaneous at 30-s scan intervals. The recoding of several behaviors was less accurate when applying longer scan intervals. Data from continuous and instantaneous sampling at 30-s scan intervals were compared with Wilcoxon test. Results showed no significant differences for posture, position in the pen and all behaviors (p > 0.05) except vocalizing (p = 0.003). The same test did not highlight significant differences due to method of dealing with out of sight for both sampling rules (p > 0.05). Correlation between continuous and instantaneous sampling were prevalently high at 30-s intervals and they decreased as the length of scan intervals increased for most behaviors. Results confirmed the first two assumptions suggesting that continuous sampling is more accurate, in particular for short and rare behaviors, and caution against the suitability of dam behavioral data collected using instantaneous sampling at scan intervals of minutes. The third assumption was not proven by this study. Results should be considered in light of the development of new technologies that relies on data acquired by sensors and imaging to monitor cow-calf welfare and behavior post-parturition.

Patterns and determinants of the global herbivorous mycobiome.

Despite their role in host nutrition, the anaerobic gut fungal (AGF) component of the herbivorous gut microbiome remains poorly characterized. Here, to examine global patterns and determinants of AGF diversity, we generate and analyze an amplicon dataset from 661 fecal samples from 34 mammalian species, 9 families, and 6 continents. We identify 56 novel genera, greatly expanding AGF diversity beyond current estimates (31 genera and candidate genera). Community structure analysis indicates that host phylogenetic affiliation, not domestication status and biogeography, shapes the community rather than. Fungal-host associations are stronger and more specific in hindgut fermenters than in foregut fermenters. Transcriptomics-enabled phylogenomic and molecular clock analyses of 52 strains from 14 genera indicate that most genera with preferences for hindgut hosts evolved earlier (44-58 Mya) than those with preferences for foregut hosts (22-32 Mya). Our results greatly expand the documented scope of AGF diversity and provide an ecologically and evolutionary-grounded model to explain the observed patterns of AGF diversity in extant animal hosts.

Host Species Affects Bacterial Evenness, but Not Diversity: Comparison of Fecal Bacteria of Cows and Goats Offered the Same Diet.

The aim of this study was to compare the diversity and composition of fecal bacteria in goats and cows offered the same diet and to evaluate the influence of animal species on the gut microbiome. A total of 17 female goats (Blond Adamellan) and 16 female cows (Brown Swiss) kept on an organic farm were fed pasture and hay. Bacterial structure in feces was examined by high-throughput sequencing using the V4-V5 region of the 16S rRNA gene. The Alpha diversity measurements of the bacterial community showed no statistical differences in species richness and diversity between the two groups of ruminants. However, the Pielou evenness index revealed a significant difference and showed higher species evenness in cows compared to goats. Beta diversity measurements showed statistical dissimilarities and significant clustering of bacterial composition between goats and cows. Firmicutes were the dominant phylum in both goats and cows, followed by Bacteroidetes, Proteobacteria, and Spirochaetes. Linear discriminant analysis with effect size (LEfSe) showed a total of 36 significantly different taxa between goats and cows. Notably, the relative abundance of Ruminococcaceae UCG-005, Christensenellaceae R-7 group, Ruminococcaceae UCG-010, Ruminococcaceae UCG-009, Ruminococcaceae UCG-013, Ruminococcaceae UCG-014, Ruminococcus 1, Ruminococcaceae UCG-002, Lachnospiraceae NK4A136 group, Treponema 2, Lachnospiraceae AC2044 group, and Bacillus was higher in goats compared to cows. In contrast, the relative abundance of Turicibacter, Solibacillus, Alloprevotella, Prevotellaceae UCG-001, Negativibacillus, Lachnospiraceae UCG-006, and Eubacterium hallii group was higher in cows compared with goats. Our results suggest that diet shapes the bacterial community in feces, but the host species has a significant impact on community structure, as reflected primarily in the relative abundance of certain taxa.

Humans and Goats: Improving Knowledge for a Better Relationship.

There is consensus that the quality of the human-animal relationship (HAR) is relevant to guarantee appropriate levels of animal welfare. Given the impact that HAR may have on both goats and human beings, the aim of the present review is to elucidate: (1) how humans and goats communicate; (2) which are the factors affecting human-goat interactions; (3) how we can measure the quality of this relationship. The systematic review led to the selection of 58 relevant articles. Effective human-goat communication takes place by means of visual, tactile and auditory stimuli and, to a less extent, via olfactory and gustative stimuli. Goats have well-developed socio-cognitive abilities and rely on humans to get relevant information. A deep knowledge of goats' communication means and socio-cognitive abilities may greatly help improving the human-goat relationship. Management practices (e.g., rearing methods, amount and quality of interactions), as well as genetic selection for suitable individual traits, may contribute to improving HAR. Several measures to assess the quality of HAR have been validated, including avoidance in the pen and at the feeding rack and latency to first contact. Finally, farmers' attitudes and empathy with goats, as well as their motivation to work with animals, should be improved through appropriate training.

Feasibility and Reliability of the AWIN Welfare Assessment Protocol for Dairy Goats in Semi-extensive Farming Conditions.

The aim of this study was to test the feasibility and reliability of the Animal Welfare Indicators (AWIN) protocol for welfare assessment of dairy goats when applied to semi-extensive farming conditions. We recruited 13 farms located in the NW Italian Alps where three assessors individually and independently applied a modified version of the AWIN welfare assessment protocol for goats integrated with some indicators derived from the AWIN welfare assessment protocol for sheep. The applied protocol consisted of nine individual-level (body condition score, hair coat condition, abscesses, overgrown claws, udder asymmetry, fecal soiling, nasal discharge, ocular discharge, and improper disbudding) and seven group-level (severe lameness, Qualitative Behavior Assessment-QBA, thermal stress, oblivion, Familiar Human Approach Test-FHAT, synchrony at grazing, synchrony at resting) animal-based indicators. On most farms, the level of welfare was good. Many of the considered welfare problems (overgrown claws, fecal soiling, discharges, and thermal stress) were never recorded. However, oblivion, severe lameness, hair coat condition and abscesses were detected on some farms, with percentages ranging from 5 to 35%. The mean percentage of animals with normal body condition was 67.9 ± 5.7. The level of synchronization during resting was on average low (14.3 ± 7.2%). The application of the whole protocol required more than 4 h/farm and 3 min/goat. The inter-observer reliability varied from excellent (udder asymmetry, overgrown claws, discharges, synchrony at resting, use of shelter) to acceptable (abscesses, fecal soiling, and oblivion), but insufficient for hair coat condition, improper disbudding, synchrony at grazing, QBA. Differences in background of the assessors and feasibility constraints (i.e., use of binoculars in unfenced pastures, individual-level assessment conducted during the morning milking in narrow and dark pens, difficulties when using the scan and instantaneous sampling method due to the high number of animals that moved at the same time) can affect the reliability of data collection. Extensive training seems necessary for properly scoring animals when applying the QBA, whereas the FHAT to evaluate the Human-Animal Relationship of goats at pasture seems promising but needs to be validated. Indicators that evaluate the synchrony of activities require to be validated to identify the best moment to perform the observations during the day.

Effect of Weeping Teats on Intramammary Infection and Somatic Cell Score in Dairy Goats.

Mastitis is the most common disease affecting dairy goats and causing economic losses. Although it is accepted that increased somatic cell count (SCC) is mainly a response to infection, its reliability for subclinical mastitis detection in goats is controversial. Indeed, many physiological and extrinsic variables can increase SCC, including breed, parity, age, stage of lactation, seasonal variations, and milking methods. In some animals, milk-secreting tissue is present in the wall of the teat and, in some instances, milk can filter through pores in the skin to the udder surface. This condition is known as "weeping teat" (WT). In these animals, mammary tissue might be prone to develop bacterial infections, although limited information is provided. Weeping teat seems to have a genetic background and is reported to be especially found in goat breeds selected for high milk production. Moreover, it is observed a genetic correlation between WT and decreased milk yield as well as increased somatic cell scores (SCS). Since information on this topic is very limited, this study aimed at investigating any possible relationship between WT, high SCC, and the presence of bacteria in goat milk. Alpine goat farms in Northern Italy were selected based on the presence of WT. Each herd was divided into two age-matched groups, identified as case (WT+) and control (WT-). Half-udder milk samples were collected aseptically at three timepoints; bacteriological analysis was performed, and SCC were determined and transformed in SCS. There was a positive association between SCS and the presence of bacteria in milk (P = 0.037) overall, whereas WT udder defect was associated with positive bacterial culture in just one herd (P = 0.053). Thus, this herd was further investigated, repeating the sampling and the analysis on the following year. The positive association between high SCS and the presence of bacteria in milk was then confirmed (P = 0.007), whereas no association with WT condition was found. These results indicate that WT defect is usually unrelated to both the outcome of milk bacterial culture and SCS. As a side outcome, we could confirm the role of bacterial infection in increasing SCS.

Evaluation of Inter-Observer Reliability of Animal Welfare Indicators: Which Is the Best Index to Use?

This study focuses on the problem of assessing inter-observer reliability (IOR) in the case of dichotomous categorical animal-based welfare indicators and the presence of two observers. Based on observations obtained from Animal Welfare Indicators (AWIN) project surveys conducted on nine dairy goat farms, and using udder asymmetry as an indicator, we compared the performance of the most popular agreement indexes available in the literature: Scott's π, Cohen's k, kPABAK, Holsti's H, Krippendorff's α, Hubert's Γ, Janson and Vegelius' J, Bangdiwala's B, Andrés and Marzo's ∆, and Gwet's γ(AC1). Confidence intervals were calculated using closed formulas of variance estimates for π, k, kPABAK, H, α, Γ, J, ∆, and γ(AC1), while the bootstrap and exact bootstrap methods were used for all the indexes. All the indexes and closed formulas of variance estimates were calculated using Microsoft Excel. The bootstrap method was performed with R software, while the exact bootstrap method was performed with SAS software. k, π, and α exhibited a paradoxical behavior, showing unacceptably low values even in the presence of very high concordance rates. B and γ(AC1) showed values very close to the concordance rate, independently of its value. Both bootstrap and exact bootstrap methods turned out to be simpler compared to the implementation of closed variance formulas and provided effective confidence intervals for all the considered indexes. The best approach for measuring IOR in these cases is the use of B or γ(AC1), with bootstrap or exact bootstrap methods for confidence interval calculation.

What's in a Meow? A Study on Human Classification and Interpretation of Domestic Cat Vocalizations.

Although the domestic cat (Felis catus) is probably the most widespread companion animal in the world and interacts in a complex and multifaceted way with humans, the human-cat relationship and reciprocal communication have received far less attention compared, for example, to the human-dog relationship. Only a limited number of studies have considered what people understand of cats' human-directed vocal signals during daily cat-owner interactions. The aim of the current study was to investigate to what extent adult humans recognize cat vocalizations, namely meows, emitted in three different contexts: waiting for food, isolation, and brushing. A second aim was to evaluate whether the level of human empathy toward animals and cats and the participant's gender would positively influence the recognition of cat vocalizations. Finally, some insights on which acoustic features are relevant for the main investigation are provided as a serendipitous result. Two hundred twenty-five adult participants were asked to complete an online questionnaire designed to assess their knowledge of cats and to evaluate their empathy toward animals (Animal Empathy Scale). In addition, participants had to listen to six cat meows recorded in three different contexts and specify the context in which they were emitted and their emotional valence. Less than half of the participants were able to associate cats' vocalizations with the correct context in which they were emitted; the best recognized meow was that emitted while waiting for food. Female participants and cat owners showed a higher ability to correctly classify the vocalizations emitted by cats during brushing and isolation. A high level of empathy toward cats was significantly associated with a better recognition of meows emitted during isolation. Regarding the emotional valence of meows, it emerged that cat vocalizations emitted during isolation are perceived by people as the most negative, whereas those emitted during brushing are perceived as most positive. Overall, it emerged that, although meowing is mainly a human-directed vocalization and in principle represents a useful tool for cats to communicate emotional states to their owners, humans are not particularly able to extract precise information from cats' vocalizations and show a limited capacity of discrimination based mainly on their experience with cats and influenced by empathy toward them.

Intrinsic and Extrinsic Quality Attributes of Fresh and Semi-Hard Goat Cheese from Low- and High-Input Farming Systems.

In this study, we investigated the lipid composition of fresh and semi-hard goat cheese produced in three Italian farms as well as the welfare assessment of goats reared in these farms. The fatty acid (FA) profile of cheese samples were found to be strictly related to the livestock system. Cheese collected from farms in which goats were allowed to graze and were fed diets with a higher forage/concentrate (F/C) ratio showed a FA profile represented by higher contents of health-promoting fatty acids. In the same samples, the health lipid indices showed the most favorable values. Conversely, cheese samples collected from a conventional-lowland farm, where goats were fed with higher amounts of concentrates and lower F/C ratio, presented a lower nutritional quality, characterized by the worst results for what concerns the health lipid indices. Then, we built a multivariate model able to discriminate samples coming from farms managed by a low-input system from those coming from farm managed by a high-input system. The comparison of animal welfare measurements and fatty acids data showed that a better intrinsic quality of low-input farms did not always correspond to better extrinsic quality, suggesting that the information on the livestock system is not always enough to provide consumers with complete awareness of the total product quality.

Deslorelin subcutaneous implants in Oryx dammah males for reproductive control.

The aim of this study was to assess the effects of the deslorelin subcutaneous implant as a temporary contraceptive method in the Oryx dammah male. For this purpose, deslorelin at different doses, i.e. 14.1 mg and 9.4 mg, was subcutaneously implanted in three males (Phase 1) and one male (Phase 2) adult Oryx dammah, respectively. Quantitative behavior evaluation and androgen concentrations in feces and plasma were assessed before and after implant application. Fecal androgen concentrations observed in treated males were compared with those measured in one orchiectomized male and two females. Fecal androgen concentrations increased up to 15 days after the implant application, then progressively decreased, reaching the basal level at day150 in Phase 1. In Phase 2, levels remained high until day 60 and returned to basal level on day 120. Plasma testosterone concentration was higher on the day of implant application than three months later, but with variable ranges among males. A general increase of activity levels and hierarchical changes were observed after treatment, in accordance with hormonal variations. Despite males cohabiting with two fertile females during the observation period, no births were recorded. However, between the end of Phase 1 and the beginning of Phase 2, i.e. about 10-11 months after the first deslorelin implant, a fertile mating occurred leading to the birth of a calf. Therefore, we can hypothesize a contraceptive effect up to 10 months after the implant. Testicular histology performed on one male at the end of the Phase 2 showed no spermatogenetic activity. Our results suggest that deslorelin implant can be used to temporarily control reproduction in the Oryx dammah male. Behavior and fecal androgen measurements were useful and repeatable, non-invasive methods to monitor response.

Welfare Assessment on Pasture: A Review on Animal-Based Measures for Ruminants.

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.

Evaluation of Pain Mitigation Strategies in Goat Kids after Cautery Disbudding.

Nowadays, most of the goat milk production in developed countries is done in intensive indoors production systems. In these systems, procedures such as disbudding are performed routinely. Disbudding is done in young goat kids and is a recognised as a painful procedure. Pain mitigation strategies have been extensively researched, but a method that is effective in mitigating pain as well as being safe and practical has not yet been found. In this paper we used three treatment groups: one control and two groups with pain mitigation strategies for cautery disbudding, one using local anaesthesia (lidocaine) and a second one using local anaesthesia (lidocaine) plus an analgesic (flunixin meglumine). The behaviour of twenty-seven goat kids was recorded for three hours after disbudding. Overall, the goat kids that received both pain mitigation treatments dedicated more time performing active and positive behaviours. Nevertheless, the incidence of behaviours related to pain and discomfort was not consistently reduced. Research is still needed to find a practical and effective pain mitigation strategy for disbudding. A solution to this challenge would improve animal welfare as well as address societal concerns linked to the suffering of farm animals.

How Can We Assess Positive Welfare in Ruminants?

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.

Automatic Classification of Cat VocalizationsEmitted in Different Contexts.

Cats employ vocalizations for communicating information, thus their sounds can carry a widerange of meanings. Concerning vocalization, an aspect of increasing relevance directly connected withthe welfare of such animals is its emotional interpretation and the recognition of the production context.To this end, this work presents a proof of concept facilitating the automatic analysis of cat vocalizationsbased on signal processing and pattern recognition techniques, aimed at demonstrating if the emissioncontext can be identified by meowing vocalizations, even if recorded in sub-optimal conditions. Werely on a dataset including vocalizations of Maine Coon and European Shorthair breeds emitted in threedifferent contexts: waiting for food, isolation in unfamiliar environment, and brushing. Towards capturing theemission context, we extract two sets of acoustic parameters, i.e., mel-frequency cepstral coefficients andtemporal modulation features. Subsequently, these are modeled using a classification scheme based ona directed acyclic graph dividing the problem space. The experiments we conducted demonstrate thesuperiority of such a scheme over a series of generative and discriminative classification solutions. Theseresults open up new perspectives for deepening our knowledge of acoustic communication betweenhumans and cats and, in general, between humans and animals.

Understanding Cows' Emotions on Farm: Are Eye White and Ear Posture Reliable Indicators?

Understanding the emotions of dairy cows is primarily important in enhancing the level of welfare and provide a better life on farm. This study explored whether eye white and ear posture can reliably contribute to interpret valence and arousal of emotions in dairy cows. The research was conducted in five Italian dairy farms. Four hundred and thirty-six photographs of cows' heads were scored (four-level), according to the eye white and ear posture during feeding, resting, pasture, and an avoidance distance test at the feeding rack (ADF test). Eye white and ear posture were significantly correlated and influenced by the context (P = 0.001). Pasture was the most relaxing context for cows (67.8% of half-closed eyes; 77.3% ears hung down or backwards). The excitement during ADF test was high, with 44.8% of eye white being clearly visible and ears directed forwards to the approaching assessor (95.5%). Housing and management mostly influenced emotions during feeding and resting (P = 0.002 and P = 0.001, respectively): where competition for feeding places and cubicles was low, the cows showed the highest percentages of half-closed eyes and ears backwards or hung down. This research supports the use of eye white and ear posture as reliable indicators of emotions in dairy cows.

Analysis of 19 Minerals and Cortisol in Red Deer Hair in Two Different Areas of the Stelvio National Park: A Preliminary Study.

The aim of the study was to perform an investigation on the concentration of 19 minerals and cortisol in red deer (Cervus elaphus) hair, a matrix that is easy to collect with non-invasive and painless sampling, able to represent an integrative values of long-term substance concentrations, and able to give useful information, also when performed on dead animals, given its extreme stability over time. In the study thirty-five animals were included, coming from two different sides of a valley in the Stelvio National Park, where official water analysis had pointed out elevated concentrations of As in one of the two orographic sides. Hair cortisol concentrations were measured using a RIA(Radio Immuno Assay), while minerals were detected using ICP-MS (Inductively Coupled Plasma- Mass Spectrometry). Results showed a negative relationship between cortisol and some mineral concentrations (Li, Co, As, Cd, Cr and Tl) and significant differences in some mineral concentrations between park areas (Al, Co, Cu, Cd and Ni). As, Cr and cortisol differences approached statistical significance. This preliminary study represents a step forward in the study of wildlife allostatic load and a valid method for applications in wildlife management programs, in environmental studies and in public health programs.

Text Mining Analysis to Evaluate Stakeholders' Perception Regarding Welfare of Equines, Small Ruminants, and Turkeys.

Welfare of animals significantly depends on how stakeholders perceive their needs and behave in a way to favor production systems that promote better welfare outcomes. This study aimed at investigating stakeholders' perception of the welfare of equines, small ruminants, and turkeys using text mining analysis. A survey composed by open-ended questions referring to different aspects of animal welfare was carried out. Text mining analysis was performed. A total of 270 surveys were filled out (horses = 122, sheep = 81, goats = 36, turkeys = 18, donkeys = 13). The respondents (41% veterinarians) came from 32 different countries. To describe welfare requirements, the words "feeding" and "water" were the most frequently used in all the species, meaning that respondents considered the welfare principle "good feeding" as the most relevant. The word "environment" was considered particularly important for turkeys, as well as the word "dry", never mentioned for other species. Horses stakeholders also considered "exercise" and "proper training" important. Goat stakeholders' concerns are often expressed by the word "space", probably because goats are often intensively managed in industrialized countries. Although the sample was too small to be representative, text mining analysis seems to be a promising method to investigate stakeholders' perception of animal welfare, as it emphasizes their real perception, without the constraints deriving by close-ended questions.

The Use of Qualitative Behaviour Assessment for the On-Farm Welfare Assessment of Dairy Goats.

This research investigated whether using qualitative behaviour assessment (QBA) with a fixed list of descriptors may be related to quantitative animal- (ABM) and resource-based (RBM) measures included in the AWIN (Animal Welfare Indicators) welfare assessment prototype protocol for goats, tested in 60 farms. A principal component analysis (PCA) was conducted on QBA descriptors; then PCs were correlated to some ABMs and RBMs. Subsequently, a combined PCA merged QBA scores, ABMs and RBMs. The study confirms that QBA can identify the differences in goats' emotions, but only few significant correlations were found with ABMs and RBMs. In addition, the combined PCA revealed that goats with a normal hair coat were scored as more relaxed and sociable. A high farm workload was related to bored and suffering goats, probably because farmers that can devote less time to animals may fail to recognise important signals from them. Goats were scored as sociable, but also alert, in response to the presence of an outdoor run, probably because when outdoors they received more stimuli than indoors and were more attentive to the surroundings. Notwithstanding these results, the holistic approach of QBA may allow to register animals' welfare from a different perspective and be complementary to other measures.

Tissue distribution and acute toxicity of silver after single intravenous administration in mice: nano-specific and size-dependent effects.

BACKGROUND: 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. METHODS: 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. RESULTS: 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). CONCLUSIONS: 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.

Feasibility and validity of animal-based indicators for on-farm welfare assessment of thermal stress in dairy goats.

This investigation tested the feasibility and validity of indicators of cold and heat stress in dairy goats for on-farm welfare assessment protocols. The study was performed on two intensive dairy farms in Italy. Two different 3-point scale (0-2) scoring systems were applied to assess cold and heat stress. Cold and heat stress scores were visually assessed from outside the pen in the morning, afternoon and evening in January-February, April-May and July 2013 for a total of nine sessions of observations/farm. Temperature (°C), relative humidity (%) and wind speed (km/h) were recorded and Thermal Heat Index (THI) was calculated. The sessions were allocated to three climatic seasons, depending on THI ranges: cold (<50), neutral (50-65) and hot (>65). Score 2 was rarely assessed; therefore, scores 1 and 2 were aggregated for statistical analysis. The amount of goats suffering from cold stress was significantly higher in the cold season than in neutral (P < 0.01) and hot (P < 0.001) seasons. Signs of heat stress were recorded only in the hot season (P < 0.001). The visual assessment from outside the pen confirms the on-farm feasibility of both indicators: No constraint was found and time required was less than 10 min. Our results show that cold and heat stress scores are valid indicators to detect thermal stress in intensively managed dairy goats. The use of a binary scoring system (presence/absence), merging scores 1 and 2, may be a further refinement to improve the feasibility. This study also allows the prediction of optimal ranges of THI for dairy goat breeds in intensive husbandry systems, setting a comfort zone included into 55 and 70.