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Accurate and reliable on-farm protocols are essential in sheep welfare evaluation. However, protocols with a high number of measures are demanding in terms of time. The aim of this trial was to use a few already validated, reliable, feasible, rapid and non-invasive animal-based indicators to evaluate and compare welfare on dairy and mixed purpose sheep farms. Indicators included prevalence of leanness, fleece condition, fleece cleanliness, skin lesions, tail docking, lameness, hoof overgrowth and mastitis. Farmers were asked to rank the relevance of the animal welfare indicators. Nine farms with dairy and nine farms with dual-purpose breeds were tested. The relevance given by eighteen farmers was affected by the type of indicators. Farmers scored mastitis as the most relevant welfare indicator, followed by leanness and lameness. The least relevant indicator was docked tail. The prevalence of poor fleece condition was lower in dairy sheep, whereas dual-purpose farms had higher prevalences of tail-docked animals. Farms showing a higher prevalence of skin lesions had animals with lower body condition and higher longevity. Hoof overgrowth was positively correlated with fleece dirtiness. No differences between dairy and dual-purpose farms were found in terms of the final score, which ranged from 33 (best welfare score) to 187 (worst welfare score). The final score was highly variable among farms. Identification of indicators showing higher prevalences allow the farmers to identify the main areas for intervention at farm level.

As different animal welfare assessment schemes are based on different assumptions but all have similar welfare claims, we studied the degree of agreement among three systems, two mainly based on resource-based measures: Animal Needs Index (ANI) and Lombardy Extension Service Index (IBS) and one mainly based on animal based-measures (i.e. Welfare Quality®). Thirty-three dairy cattle farms were assessed by one trained assessor. In order to make the welfare categories of the different welfare assessment systems comparable with one another the first three welfare categories of the ANI scheme were merged into a not acceptable category so that a score scale with 4 welfare categories was obtained. Then, we calculated the Cohen's weighted kappa (k) to assess the degree of agreement between the three welfare assessment systems in terms of welfare categories. In addition, the correlation between pair of systems in terms of total scores (the mean of the four principles of the Welfare Quality® was used for this calculation) was calculated using the Spearman correlation coefficient (rs). Our results showed that only few farms were scored in the same welfare category of different welfare assessment systems as only 12.1% of the farms were scored as enhanced and 3% of the farms were scored as acceptable according to all three welfare assessment systems. Based on k statistics, the degree of agreement in terms of welfare categories was not significant for the pairs IBS – ANI (k = 0.128; P = 0.130) and WQ – ANI (k = 0.014; P = 0.850), whereas it tended to be significant for the pair WQ - IBS (k = 0.256; P = 0.070). The Spearman correlation coefficient of the total scores confirmed the low agreement among the three systems. In particular, the correlation coefficients between ANI-IBS, IBS-WQ and ANI-WQ were rs = 0.022, 0.208 and −0,068, respectively, and none of them was significant (P = 0.905, P = 0.246 and P = 0.707, respectively). Significant correlations were only observed between the Welfare Quality® principle “Good health” and the somatic cell counts (rs = −0.500; P = 0.003), the number of lactating animals (rs = −0.360; P = 0.040), the total number of cows per herd (rs = −0.339; P = 0.053), whereas the correlation of the same principle with mortality (rs = −0.315; P = 0.074) and calving interval ((rs = −0.330; P = 0.070) tended to be significant. We conclude that different schemes based on different assessment criteria yield different results albeit making similar welfare claims.

This study aimed to assess the relationship between stockperson behavior and buffalo behavior. The research was carried out in 27 buffalo farms. The behavior of stockpeople and animals during milking and the avoidance distance at the feeding place were recorded. Recordings were repeated within one month to assess test-retest reliability. A high degree of test-retest reliability was observed for all the variables with Spearman rank correlation coefficients (rs) ranging from 0.578 (p = 0.002, df = 25) for the number of kicks performed during milking to 0.937 (p &lt, 0.001, df = 25) for the percentage of animals moving when approached by &le, 0.5 m. The number of negative stockperson interactions correlated positively with the number of kicks during milking (rs = 0.421, p &lt, 0.028, df = 25) and the percentage of animals injected with oxytocin (rs = 0.424, p &lt, 0.027), whereas the percentage of negative stockperson interactions correlated positively with the percentage of buffaloes moving when approached at a distance &gt, 1 m (rs = 0.415, p &lt, 0.031, df = 25). In a subsample of 14 farms, milk yield was correlated positively with the number of positive interactions (rs = 0.588, p &lt, 0.027, df = 12) and correlated negatively with the number of steps performed by the animals during milking (rs = &minus, 0.820, p &lt, 0.001, df = 12). This study showed that the quality of stockpeople interactions may affect buffalo behavior and production.

Simple Summary The concern for better farm animal welfare has been greatly increasing among scientists, veterinarians, farmers, consumers, and the general public over many years. As a consequence, several indicators have been developed to assess animal welfare, and several specific protocols have been proposed for welfare evaluation. Most of the indicators developed so far focus on the negative aspects of animal welfare (e.g., lameness, lesions, diseases, presence of abnormal behaviours, high levels of stress hormones, and many more). However, the lack of negative welfare conditions does not necessarily mean that animals are in good welfare and have a good quality of life. To guarantee high welfare standards, animals should experience positive conditions that allow them to live a life that is really worth living. We reviewed the existing indicators of positive welfare for farmed ruminants and identified some gaps that still require work, especially in the domains of Nutrition and Health, and the need for further refinement of some of the existing indicators. Abstract 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.

The effects of different dietary levels of maize silage (10% v. 36% DM) and group size (7 v. 14 animals) were assessed on growth performance and in vivo digestibility of 28 male fattening buffaloes. In addition, the effects of diet on meat quality and group size on behaviour and immune response were separately evaluated. Animals were weighed and assigned to three groups. The high silage - low size group (HL) was fed a total mixed ration (TMR) containing 36% DM of maize silage and consisted of seven animals (age 12.7±2.6 months; BW 382.2±67.7 kg at the start of the study). The low silage - low size group (LL) was fed a TMR containing 10% DM of maize silage and consisted of seven animals (age 13.0±2.7 months; BW 389.4±72.3 kg). The high silage - high size group (HH) was fed the 36% maize silage DM diet and consisted of 14 animals (age 13.9±3.25 months; BW 416.5±73.9 kg). Total space allowance (3.2 indoor+3.2 outdoor m2/animal) was kept constant in the three groups, as well as the ratio of animals to drinkers (seven animals per water bowl) and the manger space (70 cm per animal). Growth performance, carcass characteristics and digestibility were influenced neither by dietary treatment nor by group size, even if the group fed 36% maize silage diet showed a higher fibre digestibility. No effect of diet was found on meat quality. Group size did not affect the behavioural activities with the exception of drinking (1.04±0.35% v. 2.60±0.35%; P

In western countries buffaloes are emerging as an alternative species for dairy product differentiation. In the near future dairy enterprises will have to meet increasing environmental regulations. Life Cycle Assessment has been widely used to assess the environmental impact of different milk production systems. We aimed to examine the environmental consequences of two dairy buffalo heifer farming systems using the Life Cycle Assessment approach. The primary data were collected from 32 subjects aged 7–8 months at the start of the experiment until they reached the age of puberty in about 12 months (i.e. at the age of 19–20 months). Sixteen animals were group-housed and confined in an indoor slatted floor pen (4 m2/animal) with an outdoor paddock (4 m2/animal); 16 others free-ranged on a Mediterranean natural pasture. The environmental charges for global warming potential expressed in terms of total emissions of carbon dioxide equivalent (CO2-eq) was 35.7% less in the free-ranging system as compared with the confined system. The main source of pollution for the confined system was biogenic methane (total amount produced = 2012 kg CO2-eq) followed by CO2 from fossil fuels (total amount produced = 1006 kg CO2-eq). The environmental charges for acidification potential, eutrophication potential and non-renewable energy use were 86.3%. 60.0% and 81.4% lower in the free-ranging system compared with the confined system, respectively. In the confined system the largest pollutant in terms of acidification potential was ammonia, whereas nitrate leaching in water (total amount produced = 3311 g SO2-eq) and the use of crude oil (total amount consumed = 5684 MJ-eq) were the most relevant for eutrophication potential and non-renewable energy use, respectively. Our results represent the first example of study comparing the environmental impact of an intensive dairy farming system with an alternative natural pasture based system in the Mediterranean region and suggest that the conduction of the unproductive part of the cycle on natural pasture can promote the reduction of several sources of pollution both in atmosphere and in water. Conversely, land occupation was higher in the free-ranging system as compared with the confined system (20,349 vs 1381 m2 year, respectively). However, the software and the database used for this calculation only considered duration of land use and yield per area unit, whereas no relevance was given to the quality of land use in terms of animal welfare promotion, contribution to biodiversity conservation, and maintenance of economically active social communities. Therefore, we suggest that the estimation of the impact categories related to land occupation would include aspects concerning the nature of the land.

The aim of this paper was to examine the environmental consequences of raising buffalo calves, as assessed by life cycle assessment. Life cycle assessment has been widely used to assess the environmental impact of different livestock production systems. The primary data were collected from 32 animals aged 0-90 days. Calves were allowed to uptake colostrum before separation from their mothers within 24-h after birth. After separation, the animals were kept individually (1 x 2 m) for 8 weeks and then housed in groups of 8 in straw-bedded pens (5 x 8 m). According to the results of the analysis, the contributions from all calves to global warming potential, acidification potential, eutrophication potential, non-renewable energy use was high. In particular, the impact generated from global warm potential was 7 kg CO2 – eq per day, and the impact of non-renewable energy use was 38 MJ – eq per day. Our results estimated for the first time the environmental impact generated from buffalo calves. We conclude that the strategies to mitigate the effects on the environment impact must start from the birth of the buffalo calves and then continue throughout its life cycle.

This study investigates the stability of the milking order of cows entering into the milking parlour of a commercial dairy herd, and changes to this order consequent to a change in health status and a change in the membership of the group of cows being milked. In large herds it is difficult for stockmen to monitor all cows closely. If changes in the milking order are indicative of health or other problems, the monitoring of any changes in the order could alert stockmen to such problems. This could therefore be a tool to add to the parameters collected routinely in precision livestock farming (PLF). The milking process of 692 cows, in seven different feeding groups, over a period of six months, was monitored. The milking order was found to be stable within days, and across days, but was more variable within milking sessions. Cows with mastitis (P < 0.001) entered the parlour later than when they were healthy. Cows with metritis entered into the milking parlour earlier than usually(P < 0.05). When new cows entered a group they fitted into a stable position within the parlour entrance order within two days. There were no effects found of parity, age, days in milk, milk production or milking duration on the milking order. It is concluded that theregular monitoring of milking order, and flagging of changes in this order could be a useful tool in the early identification of the presence of disease in cows

The aim of this study was to investigate the effect pre-partum habituation in the milkingparlour on behaviour and lactation performance of buffalo heifers. Sixteen buffalo heifersin late gestation were used for this study. The animals, with an age ranging from 30 to44 months at the start of the study, were equally allocated into two treatments and bal-anced for estimated calving date. Eight animals received pre-partum habituation for 10 daysbefore the estimated calving date (group H), while the eight others received no treatmentand served as the control (group C). During the pre-partum habituation sessions, group Hanimals were moved to the milking parlour and left for 10 min in the milking stall onceeach day, where the udder was thoroughly washed with warm water, wiped with a dis-posable towel and massaged. The behaviours, registered from entrance into the milkingstall to exit, were step and kick. After calving milk flow profiles, milk yield and milk qual-ity variables were determined along with behavioural recordings. The H animals duringthe habituation procedure showed a reduction in the number of steps (P < 0.001) and kicks(P < 0.01). After calving the H animals performed fewer steps than the control animals at 0(P < 0.001), 3 (P < 0.01), 6 (P < 0.01), 13 (P < 0.01) and 20 (P < 0.01) days after calving. Animalsfrom group H also performed fewer kicks than control animals at 0 (P < 0.001), 3 (P < 0.01),6 (P < 0.01) and 13 (P < 0.01) days after calving. In the C group a reduction in the numberof steps (P < 0.001) and kicks (P < 0.001) was observed as lactation proceeded, whereas forgroup H only a tendency for a reduction over lactation was detected. Pre-partum habitu-ation did not significantly affect milk quality or milk flow variables. Milk yield in the first3 min of milking (P < 0.001), and average milk flow (P < 0.001), increased throughout theexperimental period, whereas the duration of the pre-milking phase decreased as lactationproceeded (P < 0.001). This study shows that buffalo heifers exposed to a pre-partum habit-uation programme performed fewer steps and kicks than control animals during milking.Therefore, it is concluded that using this treatment can reduce the level of restlessness inbuffalo heifers during milking.

This review deals with the behaviour of river buffaloes (Bubalus bubalis), in confinement and in extensive conditions, also focusing on the effects of different housing and rearing conditions on their welfare. The behavioural repertoire expressed by buffaloes in extensive and intensive conditions is similar to those displayed by other domestic ruminants. However, through natural selection, buffaloes have also acquired several morphological, physiological and behavioural (i.e. wallowing) adaptations to hot climatic conditions. Buffaloes kept in intensive conditions and having no access to pasture and water for wallowing extend their periods of idling and are less often involved in investigative activities. Confinement is also associated with a reduction of space; however, no specific studies have been carried out to determine the specific requirements of this species. Space restriction can adversely affect various aspects of buffalo welfare, such as health (increased levels of lesions and injuries), social behaviour (increased number of agonistic interactions) and heat dissipation. The buffaloes, originating from tropical areas, are well adapted to large variations in food availability and quality, and to dietetic unbalances. As to human animal relationship, it has been observed that the incidence of stepping and kicking behaviour of buffaloes in the milking parlour is positively correlated with the frequency of oxytocin injections, whereas the frequency of positive stockperson interactions with the animals such as talking quietly, petting and gentle touching are negatively correlated with the number of kicks during milking. Data from farms where both dairy cattle and buffaloes are present show that avoidance distance measured in the pen is lower in buffaloes than in cattle. This may be attributed to the fact that buffaloes are generally recognised to be curious animals. Finally, the effects of different farming practices on animal-related indicators are described. However, these measures should be integrated into a monitoring protocol, such as the Welfare Quality® scheme, to reliably assess buffalo welfare in the current intensive farming conditions.

Validity and reliability of simple measures of human animal relationship, such as avoidance distance in the pen (AD), avoidance distance at the manger (ADM) and percentage of animals not accepting to be touched at the manger (ADM0), were assessed in 20 sheep farms. Validity was tested using an attitude questionnaire administered to the stock-people and calculating the correlation of questionnaire scores with AD, ADM and ADM0. Convergence among these three variables was also computed. Short-term test–retest reliability was evaluated correlating the measurements recorded in the two visits performed in each farm. Stock-people rated the four sections of the questionnaire at scores above the central point, thus indicating that the attitude of stock-people was generally positive. AD and ADM were higher than reported in other dairy ruminant species. AD was significantly higher than ADM (P < 0.001). A high correlation was observed between ADM and ADM0 (r = 0.810; P < 0.001), whereas lower correlations were observed between AD and ADM (r = 0.445; P < 0.05) and AD and ADM0 (r = 0.570; P < 0.01). ADM and ADM0 showed significant correlation with the questionnaire total score (r = −0.435 and −0.509, respectively; P < 0.05), whereas AD was unrelated with the questionnaire total score. A good test–retest reliability was observed for all the three measures (P < 0.001), although ADM0 showed a lower coefficient value (r = 0.679) in comparison with AD (r = 0.898) and ADM (r = 0.845). The marked gregarious behaviour exhibited by sheep in enclosed areas (when approached in the pen animals may run towards the observer) may impair AD feasibility in terms of observer safety. The present results indicate the measures gathered at the manger as the most suitable for the assessment of human–animal relationship in sheep.

Welfare Quality® is the acronym of an European research project focussing on the integration of animal welfare in the food quality chain: from public concern to improved welfare and transparent quality. The project aims to accommodate societal concerns and market demands, to develop reliable on-farm monitoring systems, product information systems, and practical species-specific strategies to improve animal welfare. Forty-four institutes and universities, representing 20 countries participate in this integrated research project concerning 3 animal species (cattle, pigs and chicken). From 2007 dairy buffalo has been also included in the list. Welfare Quality® has been co-financed by the European Commission, within the 6th Framework Programme. In this paper the development of a monitoring system to assess buffalo welfare is described.

A study was carried out at an organic buffalo dairy farm in order to investigate the effect of feeding protein peas (Pisum sativum L.) as an alternative protein source for buffalo cow diets. Two concentrates were formulated to contain (as fed basis) either 350 g/kg of soybean cake (SC) or 450 g/kg of peas (PC) as the main protein sources. The two concentrates were formulated to be almost isonitrogenous (on average crude protein 240 g/kg DM). Two groups of 10 buffalo cows were used in a 100-day lactation study (from 10 days in milk onwards). Cows were blocked into two groups according to lactation number and previous milk yield and were assigned to one of two dietary treatments: control group was offered in the milking parlour 3 kg of SC, while treatment group was offered the same quantity of PC. All cows were fed a total mixed ration containing 3 kg of SC. Daily milk yield was not affected by treatment, as well as, milk fat and protein percentages, somatic cell count, urea content and fatty acid composition.

The present study was undertaken to evaluate the inter-observer reliability of a welfare monitoring scheme to be applied to sheep, and compare the welfare state of the animals between 10 organic and 10 conventional sheep farms. Two trained observers performed recordings. A graded point protocol was used, that relies on five sheets mostly derived by the Animal Needs Index, which is mainly based on resource-based parameters. Therefore, in the fifth sheet animal-based parameters, deemed relevant to sheep welfare, were taken into account. In particular, the following animal-based variables were assessed: integument alterations, animal dirtiness, hoof overgrowth, lameness and lesions, which where scored on the basis of their prevalence (number of affected animals/numbers of observed animals), longevity (age in years), and mutilations, such as de-horning and caudotomy, evaluated in terms of presence/absence. No significant differences were observed between organic and conventional farms in terms of ANI scores, housing characteristics and animal-based parameters. This result was not surprising, as most of the farms, both conventional and organic, based their farming systems on an extensive use of the land by grazing animals. The monitoring protocol proved to be feasible (the mean time needed to perform the assessment of welfare was 85 min per farm) and reliable: a significant Spearman’s correlation coefficient between observers was observed for total score and all sheets. As to animal-based parameters, body condition could not be assessed visually due to the presence of flee in winter; the correlation between observers was significant for integument alterations, animal dirtiness, hoof overgrowth and lameness, whereas inter-observer reliability was not significant for lesions. This result indicated that more training is needed for the assessment of lesions in order to increase the reliability of the measure. In addition, we suggest visiting farms in early summer, soon after shearing, in order to make easier the detection of lesions and the assessment of body condition.

Soon after parturition a lasting and mutual ewe–lamb bond is established. However, in an increasing number of intensive sheep farms, lambs are separated from the dam at an early age. When artificial rearing is applied lambs are often kept with mothers for 2 days to allow the ingestion of maternal colostrum and then abruptly removed from their dams. Thus, lambs experience a marked emotional stress represented by the loss of the most relevant social model at this early stage of their behavioural development and a nutritional stress represented by the transition from maternal milk to a commercial milk substitute. These animals when exposed to open field tests show reduced levels of vocalization, are slower to initiate movements, spend less time in ambulatory behaviour and display an increased cortisol response than non-separated animals. In addition, artificial rearing performed on lambs from 2 days of age onward can cause decreased cellular and humoral immune responses. The main oral abnormal behaviour performed by artificially reared lambs is represented by sucking the navel or the scrotum of pen mates. This activity is evident from the initial days on reconstituted milk and lasts until weaning from milk. Attempts have been made to reduce the detrimental effects of early separation. Some of them mainly focus on the emotional aspects (it is recommended not to leave a lamb alone for artificial rearing), others aim at reducing the nutritional impact of artificial rearing (milk intake can be increased by offering a mix of ewe milk and a milk replacer during the first week and then gradually moved to a diet based only on milk substitute which results in higher growth rates). As compared with artificial rearing, early weaning performed at 3 months of age is associated with a later disruption of the mother–young bond and the consequent direct replacement of maternal milk by solid food. However, when they are given the chance, ewes and their lambs form long-term social associations which exceed the age of natural weaning, regarded as the end of the milk feeding period. Early weaned lambs emit an increased number of high pitched bleats immediately after weaning than before and this increment is still evident 2 days afterwards. Neither partial nor gradual separation from mothers is able to reduce the stress associated with early weaning. In conclusion, premature separation from mothers has clear and marked detrimental effects on various functions in lambs. For lambs maternal deprivation seems to be worse at 2 days (artificial rearing) than at 3 months of age (early weaning).

The responsiveness of 10 horses and 10 ponies to environmental challenge (represented by an open field test) was assessed using a qualitative approach based on free choice profiling methodology (FCP), which gives observers complete freedom to choose their own descriptive terms. Data were analysed with generalised Procrustes analysis (GPA), a multivariate statistical technique associated with FCP. A cross-validation of the outcomes of this approach to data recorded through quantitative behaviour analysis, and through a questionnaire given to the animals’ owner/riding instructor, was also performed using principal component analysis (PCA). Twelve undergraduate students generated their own descriptive vocabularies, by watching 20 horse/pony video clips lasting 2.5 min each. GPA showed that the consensus profile explained a high percentage of variation among the 12 observers, and differed significantly from the mean randomised profile ( p < 0.001). Two main dimensions of the consensus profile were identified, explaining 60% and 5.2% of the variation between animals, respectively. The 12 observer word charts interpreting these dimensions were semantically consistent, as they all converged towards the same meaning, albeit using different terms. The most used term to describe the positive end of axis 1 was ‘‘quiet’’, whereas ‘‘attentive’’ was the best positive descriptor of axis 2. The most frequently used descriptors for the negative ends of axes 1 and 2 were ‘‘nervous’’ and ‘‘bored’’, respectively. Thus, axis 1 was labelled as ‘‘quiet/nervous’’ and axis 2 was named as ‘‘attentive/bored’’. A marked effect of animal category was observed on the scores of the animals on the first dimension ( p < 0.001). Horses received significantly higher scores, and were thus assessed as more quiet and calm, than ponies. Conversely, ponies tended to receive lower scores on the second dimension ( p < 0.12), therefore they appeared less curious and attentive. The results of the PCA showed that the variables from different types of measurement clearly had meaningful relationships. For instance, the variables with the highest loading on the positive end of axis 1 were all indicative of tractable and docile animals, whereas axis 2 showed high loadings on the positive end for variables indicating attentive animals. Qualitative behaviour assessment proved to be an appropriate methodology for the study of horse behavioural responsiveness, in that it provided a multifaceted characterisation of horse behavioural expression that was in agreement with other quantitative and subjective assessments of the animals’ behaviour.

In order to contribute to the improvement of the national greenhouse gas emission inventory, this work aimed at estimating a country-specific enteric methane (CH4) emission factor for the Italian Mediterranean buffalo. For this purpose, national agriculture statistics, and information on animal production and farming conditions were analysed, and the emission factor was estimated using the Tier 2 model of the Intergovernmental Panel on Climate Change. Country-specific CH4 emission factors for buffalo cows (630 kg body weight, BW) and other buffalo (313 kg BW) categories were estimated for the period 1990–2004. In 2004, the estimated enteric CH4 emission factor for the buffalo cows was 73 kg/head per year, whereas that for other buffalo categories it was 56 kg/head per year. Research in order to determine specific CH4 conversion rates at the predominant production system is suggested.

The feeding management, milk yield and milk composition were investigated in two adjacent buffalo farms, one organic certified (on average, 220 lactating buffalo cows) and one conventional (on average, 314 lactating buffalo cows) located in the Sele Plain (southern Italy). Milk samples from the two farm were collected twice a month during the period from June to November 2006. Milk production was also recorded. The investigated milk components were the content of protein, fat, lactose, urea and the number of somatic cells. The following features about the feeding management of lactating buffalo cows were recorded during monthly farm visits: feed used, herbage utilization, ration composition. Fat and protein correct milk yield was higher in conventional farm than in organic one, but milk components were similar between the farms. The greatest differences were found in the somatic cell count, lower in organic milk than in conventional one, and the urea content, which was higher in organic milk than in conventional farm, but still within the normal range reported for buffalo milk.

Within the general aim of developing a Welfare Quality system for monitoring dairy buffalo welfare, this study focused on prevalence and interobserver reliability of the animal-related variables to be included in the scheme. As most of the measures were developed for cattle, the study also aimed to verify their prevalence for buffaloes. Thirty animal-based measures (22 clinical and 8 behavioral measurements) and 20 terms used for qualitative behavior assessment were assessed in 42 loose-housed buffalo farms. All farms were located in central-southern Italy. Two assessors were used (1 male and 1 female). The time needed to record all measures (animal-, resource-, and management-based) was 5.47 ± 0.48 h (mean ± SD). Interobserver reliability of animal-based measures was evaluated using Spearman rank correlation coefficient test (rs). If 0.7 is considered as threshold for high interobserver reliability, all animal-based measures were above this level. In particular, most of the coefficients were above 0.85, with higher values observed for prevalence of animals that can be touched (rs = 0.99) and prevalence of animals with iatrogenic abscess (rs = 0.97), whereas lower coefficients were found for the prevalence of vulvar discharge (rs = 0.74) and dewlap edema (rs = 0.73). Twelve out of the 20 terms used for the qualitative behavior assessment reached a satisfactory interobserver reliability (rs = 0.65). Principal component analysis of qualitative behavior assessment scores was conducted for each assessor. Both principal component 1 and principal component 2 showed high interobserver reliability (rs = 0.80 and 0.79, respectively). In addition, relevant proportions of animals were affected by welfare issues specific to buffaloes, such as overgrown claws (median = 34.1%), withers hygroma (median = 13.3%), and vulvar or uterine prolapse (median = 9.3%). We concluded that most of the investigated measures could be reliably included in the final scheme, which can be used as such to monitor buffalo welfare. However, to inform consumers about the welfare status of the animals, the data should be integrated into a single overall assessment of animal welfare, as already performed in the Welfare Quality project for dairy cattle.

In order to develop a method suitable for a continuous qualitative evaluation of animal behaviour while it changes during the observation period, two recently described techniques, the Qualitative Behaviour Assessment (QBA) and the Temporal Dominant Behavioural Expression (TDBE), derived by the Temporal Dominance of Sensations, were used in combination and applied to 8 Mediterranean buffalo heifers. Animals were subjected to two isolation tests lasting 150 s each. One consisted in isolating individual animals from the rest of the group in the indoor part of the home pen, the other in leading animals individually to an outdoor paddock, which was novel to the animals. The behaviour of the animals was video-recorded and the resulting 16 video clips were assessed by a twelve-member trained panel with previous experience in animal behaviour observation.The TDBE allowed the QBA to be performed continuously (C-QBA) during the observation period on a pre-determined list of six behavioural descriptors. Data were subjected to analysis of variance using observer (n = 12), replication (n = 4), animal (n = 16) and the first order interactions as factors. Changes of behavioural expressions during the test were assessed by dividing it into three intervals of 50 s and using the 2 one-sample test to check the dominance of each descriptor in each interval. The Wilcoxon test was used to compare quantitative and qualitative behavioural variables as assessed indoors and outdoors. The low level of variance explained for each descriptor by the interactions animal × replication and animal × assessor as compared with the animal (the F of the factor animal was roughly 10 times higher than that of the interactions), showed the intra- and inter-observer reliability of the panel. Indoors, the dominant descriptors of buffalo behaviour (calm, apathetic and curious) were all indicative of a low level of arousal and frequently switched from one another. Outdoors, most of the animals were described with terms indicating a high level of arousal (e.g. Active and Nervous) and only at the end of the test they were generally scored as Curious. Accordingly, the dominance of the descriptor Curious changed during the test when the animals were tested outdoors (2= 7.00, P < 0.05), whereas no significant changes were observed for all the other descriptors both indoors and outdoors (P > 0.05).The results indicate that C-QBA, if combined with the appropriate tools (i.e. TDBE), may be able to follow and describe the variations of animal behavioural expressions in time.

On two farms, three milking groups of buffalo cows were used to assess the consistency of entrance order and the preference for one side of the milking parlour. On Farm 1 (F1) all animals were primiparous (n=57). On Farm 2, three primiparous, 16 secondiparous and 36 multiparous cows (range 1–8) constituted group F2G1; whereas group F2G2 had 12 primiparous, 10 secondiparous and 14 multiparous cows (range 1–10). Animals were milked in auto-tandem milking parlours (2×5 and 2×6 for Farms 1 and 2, respectively). For each cow, entrance order into milking parlour, side where she was milked, milk yield, time and duration of milking were recorded. These data were derived from the computerised identification of cows. The sequence in which the cows entered the milking parlour ranged from 1 to 57 for group F1, from 1 to 55 for group F2G1 and from 1 to 36 for group F2G2. The analysis of data was conducted on 130, 120 and 92 consecutive milkings for groups F1, F2G1 and F2G2, respectively. Kendall's coefficients of concordance showed a strong constancy of the entrance order into milking parlour for groups F1 (W=0·658; χ2=4792·81; PW=0·779; χ2=5046·81; PW=0·624; χ2=2030·48; Prs=−0·221 and rs=−0·215; Prs=−0·265; Prs=0·334; Prs=−0·319; Prs=−0·325; P

The effects of dietary inclusion of Saccharomyces cerevisiae culture on intake, in vivo digestibility, and fecal nitrogen excretion were examined in dairy buffalo. Forty lactating buffalo cows were equally divided into Control and Saccharomyces groups, balanced for milk production, parity, and days in milk. Two subsequent 16-d experimental phases were carried out. For both groups during the first experimental period a TMR based on maize silage (maize-TMR) was used, whereas in the second period an alfalfa haylage (alfalfa-TMR) was administered to the animals. In each experimental period, Saccharomyces group was supplemented with 50 g/head/day of yeast (Biocell®, Limena, Padova, Italy), corresponding to 20×10 9 CFU/head/day Saccharomyces cerevisiae NCYC Sc47 strain. The yeast supplement was top-dressed onto the morning feed. Dry matter intake (DMI) was assessed for 6 consecutive d on group basis, by the difference between feed offered and refused. In the last 3 days of experimental period in vivo digestibility was determined by using acid-insoluble ash (AIA) as an intrinsic digestibility marker. Saccharomyces supplemented buffalo cows presented greater DMI of maize-TMR, whereas no statistical differences between the groups were observed for alfalfa-TMR. Saccharomyces supplementation significantly improved in vivo digestibility of both TMR. Fecal nitrogen excretion was significantly reduced by the use of yeast supplementation. Results suggest that the inclusion of Saccharomyces cerevisiae culture in the diet for lactating buffaloes can be recommended for its effects on cow’s digestive efficiency and fecal nitrogen excretion.

Twenty lactating buffalo cows, organically farmed, were used to examine the effects of including peas in total mixed ration. Two concentrates were formulated to contain, as the main protein sources, either 350 g/kg of soybean cake (CC) or 450 g/kg of peas (ExpC). Cows were blocked into two groups according to parity and previous milk yield and were assigned to one of two dietary treatments: one group was fed a diet with 6 kg/d of CC, whereas the treatment group was fed diet in which 3kg/d of CC were replaced by an equal quantity of ExpC. Digestibility of the diets and milk production of the cows were measured. The experimental period covered the whole lactation period. No differences were observed between groups for milk yield and composition, and for total tract apparent digestibility. The main hypothesis tested, that the replacement of soybean cake with peas in buffalo diet would not affect milk yield and composition, was confirmed. This suggested that the partial substitution of soybean cake with peas in diet for buffalo cows can be possible without affect performances.

The influence of artificial rearing at different ages was evaluated in Comisana lambs. Animals were assigned to three test groups (2d, 15d and 28d) on the basis of age (2, 15 or 28 days, respectively) at the time of their separation from the ewe, and three corresponding control non-separated groups (2c, 15c and 28c). On the day of separation from their dam, animals were injected i.m. with 250 μg of keyhole limpet hemocyanin in complete Freund's adjuvant. Antibody titers and surface leukocyte antigens were determined at weekly intervals. Group 2d showed a lower antibody response than the corresponding control group (F = 8.0, P < 0.01), whereas groups 15d and 28d did not differ from the control. No significant difference between test and control groups was found in the proportion of cells bearing different surface antigens. Furthermore, lambs were subjected to a behavioral test (isolation from tactile and visual contact with other animals). Younger animals showed reduced duration of movement (162.8 vs. 272.1 and 221.1 s for 2d, 15d and 28d, respectively; U = 21.0, P < 0.01 and U = 30.5, P < 0.05, respectively; Mann-Whitney U test) and enhanced latency time (15.8 vs. 3.4 and 2.3 s for 2d, 15d and 28d, respectively; U = 28.0, P < 0.01 and U = 14.5, P < 0.001, respectively; Mann-Whitney U test) in comparison with older animals. Cortisol concentration was evaluated immediately before the separation from the mother and 15, 45, 90, 150, and 225 min after. An increased cortisol level was observed in group 2d 15 min after removal from the dam (LSD: P < 0.001), whereas the other two groups did not show any significant increase. In addition, during the first 28 days of age all the animals were weighed at birth and at weekly intervals. When the animals were removed from the mother either at 2 or 15 days of age, the daily weight gain decreased significantly in comparison with the corresponding control groups (107.1 vs. 157.1 g for 2d and 2c, respectively, F = 4.8, P < 0.05; 37.3 vs. 126.8 g for 15d and 15c, respectively, F = 5.8, P < 0.05). In conclusion, early separation from the ewe affects humoral immune response and post-separation performance of lambs as a possible consequence of a reduced ability of young animals to cope with emotional and nutritional stresses.

The possibility of inducing conditioned taste aversion (CTA) by injection of apomorphine in 24 young Maltese goats and the effect of social models on the feeding behaviour of conditioned animals were evaluated. The experimental feeds were oats seed (OS) and rabbit alfalfa pellets (RP). Animals were divided into six social groups (S), including conditioned and unconditioned subjects, and two non-social groups (NS), with conditioned animals only. Within the S groups each conditioned animal stood between two unconditioned subjects while eating. When feeds were paired with apomorphine (APO feeds), no significant differences were found in consumptions between conditioned animals of the S and NS groups (P > 0.05). Unconditioned animals of the S groups did not modify their feed intakes and were not able to stimulate the intake of the APO feeds in the conditioned subjects. Consumption of the APO feeds was significantly lower than that of the control feeds (P < 0.001). RP was always consumed more than OS, even when it was associated with apomorphine, probably because of its physical similarity to the pellet of the base diet. In the two-choice test 16 out of 24 goats chose the control feed first and eight the feed associated with illness. When they chose the latter, it was RP seven out of eight times and OS only once (P < 0.01). The amounts of the consumed control feeds were significantly higher than those of the APO feeds (P < 0.01). The significant reduction in consumption of the APO feeds in comparison with the control feeds showed that goats can learn to avoid a feed that immediately produces illness. The conditioned animals did not modify their feed intakes following the influence of the unconditioned subjects.

This study applies qualitative behaviour assessment (QBA) for the first time to dairy buffaloes, using three groups of observers with different cultural backgrounds and different levels of experience in animal behaviour observation and buffalo farming. Eight buffalo heifers aged 16–18 months were subjected to two isolation tests, one performed in the indoor part of their home environment, and one in a novel outdoor paddock. Animals were filmed individually for 2.5 min, and the resulting 16 video clips were shown to three observer panels, consisting of 11 applied animal behaviour scientists from 6 European countries, 11 Italian animal scientists with a background in buffalo farming but no experience in behavioural observation, and 14 Italian undergraduate animal science students with no particular experience. A free choice profiling method was used to instruct observers in QBA, and data for the three panels were analysed separately using Generalised Procrustes Analysis. All three panels showed significant inter-observer agreement (p < 0.001) and generated two main consensus dimensions characterised as ‘calm-agitated’ and ‘curious-shy’. There were significant correlations between buffalo scores provided by each of the three observer panels on both these dimensions (dim1: Kendall W = 0.96, n = 3, χ2 = 43.28, p < 0.001; dim2: W = 0.68, n = 3, χ2 = 30.73, p < 0.01). Buffaloes viewed in the familiar indoor pen were assessed by all three panels as more calm and less agitated (dimension 1) than animals viewed in the novel outdoor pen (Wilcoxon z = −2.52, p < 0.01, z = −2.52, p < 0.01, z = −2.38, p < 0.01 for Panels 1, 2, and 3, respectively). Scores on dimension 1 for the same animals viewed in either indoor or outdoor pen were correlated at r = 0.60 (p < 0.10), 0.74 (p < 0.05) and 0.71 (p < 0.05) for Panels 1, 2, and 3, respectively. Quantitatively, buffalo in the outdoor pen displayed longer bouts of running and higher frequencies of sniffing (both p < 0.05) than those in the indoor pen. Principal component analysis showed meaningful associations between qualitative and quantitative assessments, allowing qualitative dimensions to play a valuable role in interpreting the animals’ state. The main outcomes of this study are that QBA can be usefully applied to scientific studies of dairy buffalo, and that substantial differences in observer background do not appear to diminish the reliability of QBA.

A first experiment was carried out to verify whether a goat may label her kid through licking and nursing in order to discriminate between her own and two alien kids. Furthermore, it was assessed how long after birth a kid needs to be along with its mother in order to be rejected by another goat and to be recognized by its own dam. All the 18 goats tested at 2 h accepted their own kid, whereas ten of them rejected both aliens; all the 20 goats tested at 4 h accepted their own young, whereas 17 rejected the alien isolated kid and 18 refused the alien labelled kid. In a second experiment physical contact, licking and nursing were prevented. All the 25 maternal goats accepted their own kids, whereas 11 of them rejected the alien. These two studies indicate that when the goat is left undisturbed with her kid after kidding, maternal selectivity is developed within 4 h.

The effect of dietary supplementation with Enterococcus faecium strain SF68 (E. faecium SF68) on growth performance, fecal consistency, in vivo digestibility and immune response in buffalo (Bubalus bubalis) calves was evaluated at a commercial dairy location. Forty calves were randomly assigned at 10 days of age to one of four treatments: A—milk replacer with no additive, B—milk replacer supplemented with 0.17 g/L of viable (2 × l09 cfu/g) E. faecium SF68 culture for 3 consecutive d at 7 day intervals for 11 wks, C—milk replacer supplemented with E. faecium SF68 daily for 4 wks, D—milk replacer supplemented with E. faecium SF68 daily for 11 wks. A total mixed ration (TMR) was offered ad libitum from the 5th wk of the experimental period onward. Fecal scoring was conducted twice weekly, and dry matter intake (DMI) was measured weekly. Body weight was recorded every 2 wks. Phytohemagglutinin was used to perform a skin test based on non-specific delayed type hypersensitivity, whereas keyhole limpet hemocyanin was injected subcutaneously to evaluate humoral immune response. Blood samples for the evaluation of interferon-γ were collected from the jugular vein in vacuum tubes with heparin as an anticoagulant. At the end of the experimental period, nutrient in vivo digestibility was measured by using acid-insoluble ash as an internal indigestibility marker. Probiotic treated calves exhibited better fecal consistency and higher average daily gain as compared to control calves. The use of E. faecium SF68 had no effect on TMR DMI or total tract digestibility. Immune responses and interferon γ concentration were not affected by treatment. E. faecium SF68 supplementation did not dramatically change the health of animals although it favorably affected the status of the gastro-intestinal tract. Dietary supplementation of buffalo calves with E. faecium SF68, either concentrated early after birth (i.e. 4 wks), or dispersed over 11 wks (i.e. 3 consecutive days at weekly interval), allowed the effect of the probiotic to be achieved at lower cost.

From February to August 2007 the effect of rearing system (confined (C) v. free ranging (FR)) and season (spring v. summer) was evaluated on behaviour, immune and blood parameters and beef production (experiment 1). From February to August 2008, the effect of rearing system was only evaluated on beef production (experiment 2). A total of 12 Podolian young bulls were used each year. They were slaughtered at 18 and 15 months of age in 2007 and 2008, respectively. Herbage mass and pasture composition were monitored during the 2 years. Pasture availability, in terms of herbage mass and composition, as well as its seasonal development, was similar in the 2 years. In the first experiment, FR animals spent more time walking (P < 0.05), feeding (P < 0.001) and standing (P < 0.01) and showed a lower number of agonistic (P < 0.05) and non-agonistic social interaction than C animals (P < 0.01). Significant lower concentrations of serum urea nitrogen (P < 0.001) and creatinine (P < 0.10) in FR animals indicated a lower protein nutritional status due to inadequate protein availability at pasture. As a consequence, average daily gains (P < 0.05), slaughter weight (P < 0.05) and body condition score (P < 0.01) were lower in grazing animals as compared with C bulls. Cellular immune responsiveness was higher in FR animals (P < 0.05). Similarly, antibody titre to keyhole limpet hemocyanin was higher in FR bulls at the 2nd and 3rd month after antigen injection (P < 0.05), whereas it tended to be higher at the 4th month (P < 0.10). In both experiments, grazing negatively affected meat colour in terms of lightness. Eighteen-month-old bulls also showed lower final weight, weight gain and body conditions when kept outdoor: a possible consequence of nutrient deficits, as suggested by the metabolic status of FR animals. The same animals, however, benefited from FR in terms of natural behaviour expression and immune responsiveness. When the experiment was replicated the subsequent year, on animals slaughtered at 15 months of age, no differences between the performances of FR and C animals were detected. The earlier slaughter age system was also proportionally less dependent on external inputs as grazing was not extended to the dry season when herbage mass availability was lower.

To verify the effect of 2 housing systems (with and without a pool and an ample outdoor lot) on behavior and milk yield, 45 lactating buffalo cows were group-housed in a free stall open-sided barn with concrete floor where they received 10 m(2)/head as space allowance (group NP); 43 cows were group-housed in a similar barn, but had access to an outdoor yard (36 m(2)/head) and a concrete pool of 208 m(2) (group WP). Animals were subjected to 8 sessions of instantaneous scan sampling at approximately 10-d intervals. Behavioral variables were expressed as proportions of subjects observed in each category of posture and activity. In addition, rapid behaviors such as agonistic, social, and reproductive interactions, social licking, and self-grooming were recorded continuously. These variables were expressed as number of interactions per animal. At the end of each hour of observation, temperature and relative humidity were recorded. In WP the proportion of animals observed wallowing was 0.476 +/- 0.034, whereas lower proportions were observed standing (0.389 +/- 0.029) or lying (0.141 +/- 0.021) outside the pool. In NP the proportions of animals observed standing and lying were 0.452 +/- 0.042 and 0.548 +/- 0.042, respectively. A significant relationship between mean temperatures recorded on observation days and proportion of animals in the pool was observed (r(s) = 0.41). Fewer animals from group WP were observed idling compared with buffaloes from group NP (0.44 +/- 0.024 vs. 0.509 +/- 0.024, respectively), whereas more WP animals were involved in investigative activities than NP cows (0.099 +/- 0.009 vs. 0.042 +/- 0.009, respectively). A greater number of social interactions (sniffing and nuzzling) and social lickings were observed in group WP than in group NP (0.120 +/- 0.010 vs. 0.067 +/- 0.010, and 0.151 +/- 0.018 vs. 0.090 +/- 0.018, respectively). The WP buffalo cows had a greater milk yield than NP cows (11.73 +/- 0.31 vs. 10.78 +/- 0.28 kg/d, respectively), whereas no differences between groups were observed for protein (4.86 +/- 0.04 vs. 4.80 +/- 0.03% for WP and NP, respectively) and fat contents (8.49 +/- 0.14 vs. 8.38 +/- 0.13% for WP and NP, respectively). We conclude that the provision of a pool and an ample outdoor paddock can have beneficial effects on welfare and milk production of buffaloes.

The present paper addresses the issue of buffalo welfare. Firstly, the biological characteristics and behavioural needs of buffalo are considered. Subsequently, the effects of intensive farming and some animalrelated indicators, to be used for a monitoring scheme of buffalo welfare at farm level, are described. The attention was focused on the following indicators: excessive thinning or fattening assessed with Body Condition Score (BCS) systems; cleanliness (the presence of mud may be considered positively, whereas a thick and compact layer of dung may be regarded negatively); health status (lameness, hoof overgrowth, injuries, etc.); social, aggressive, oral abnormal behaviours; animal-human relationship (avoidance distance at manger); positive indicators (qualitative assessment of behaviour, etc.); housing factors. The indicators are discussed on the basis of their validity (meaningful with respect to animal welfare), reliability (reflecting the tendency to give the same results on repeated measurements) and feasibility (concerning time and money consumed). For some aspects, the differences between buffalo and dairy cattle are also highlighted.

An Aspergillus oryzae fermentation extract and a Saccharomyces cerevisiae culture product were added to a pelleted calf starter, respectively, at 6.0 and 26 g/kg of dry matter (DM) to determine effects on faecal score, DM intake, body weight (BW) gain and in vivo digestibility. Treatments were control groups with the control starter alone (CSt) or with free access to ryegrass hay (CStH) and combinations with the fungal supplemented starter (ExpSt and ExpStH). Forty buffalo calves (12 male, 28 female; 10 calves per treatment) were started on the experiment at 10 days of age and for 12 weeks. Faecal scoring was conducted twice weekly, and DM intake was measured weekly. Every 2 weeks, BW gain was recorded and grab faecal samples were collected to examine for the presence of intestinal parasites and Salmonella. At the end of the experimental period, on a subset of 20 calves (i.e., five per treatment), nutrient in vivo digestibility was measured by using acid-insoluble ash as internal indigestibility marker. Starter DM intake was unaffected by fungal supplementation, but inclusion of fungal additives in the starter resulted in increased apparent total tract digestibility, regardless of the presence of hay in the diet. Calves fed ExpStH diet versus the CStH and ExpSt diets had improved faecal consistency, higher average daily gain and higher total tract digestibility of fibre. Supplementation of starter with A. oryzae fermentation extract and S. cerevisiae culture may improve calf digestive efficiency if forage is in the diet.

SUMMARY In the last decade, there has been an increased awareness that secondary metabolites produced by marine diatoms negatively impact the reproductive success of their principal predators, the copepods. Several oxylipins,products of the enzymatic oxidation of fatty acids, are produced when these unicellular algae are damaged, as occurs during grazing. In the past, the dinoflagellate Prorocentrum minimum, which does not produce the oxylipin 2-trans,4-trans-decadienal (DD), has been used as a live carrier to calculate daily ingestion rates of this molecule by copepod crustaceans. However, since the interaction between oxylipins and live carriers is unknown, the question as to how much and for how long ingestion of these molecules affects copepod reproduction remains a critical point to understanding the functional role of such compounds at sea. In the investigation presented here we used giant liposomes (∼7 μm) as a delivery system for the oxylipin DD, prepared in the same size range as copepod food and containing known amounts of DD. The aim of this work was to relate the ingestion of DD to the reproductive failure of the copepods Temora stylifera and Calanus helgolandicus. Liposomes were very stable over time and after 10 days of feeding, liposomes encapsulating DD reduced egg hatching success and female survival with a concomitant appearance of apoptosis in both copepod embryos and female tissues. Concentrations of DD inducing blockage were one order of magnitude lower that those used in classical feeding experiments demonstrating that liposomes are a useful tool to quantitatively analyze the impact of toxins on copepods.

From November 2005 to October 2006, sixteen buffalo heifers were used to evaluate the effect of season on grazing behaviour. They grazed a fenced Mediterranean maquis of ~40ha. Approximately 10% of this area was covered by woodland and shrub vegetation and the rest of the area by grassy habitat. From December to March, animals were supplemented with dehydrated beet-pulps and meadow hay administered in the barn. Behavioural data were collected from March to October. During a 6-h period the behaviour of a focal animal, chosen at random, was continuously monitored. All observations were conducted during daylight (between 05.30 and 15.00). During the behavioural observations, the ingestion of woody or shrub vegetation was rarely observed. The proportion of time spent grazing was not affected by season (0.600, 0.409, and 0.434, for spring, summer and autumn, respectively). Animals lay more in summer and autumn than in spring (P

In order to verify the effect of a pool on behaviour and milk yield, eighty-eight lactating buffalo cows equally allocated to two treatments were used. Group WP was provided with a concrete pool of 208 m2, 36 m2/ head + 10 m2 of outdoor and indoor space allowance, respectively, whereas group NP only received 10 m2. Less animals from group WP were observed idling (P

SUMMARYGiant liposomes are proposed as a potential delivery system in marine copepods, the dominant constituent of the zooplankton. Liposomes were prepared in the same size range as the food ingested by copepods (mean diameter of about 7 μm). The encapsulation of a hydrophilic and high molecular mass fluorescent compound, fluorescein isothiocyanate-dextran (FitcDx), within the liposomes provided a means of verifying copepod ingestion when viewed with the confocal laser-scanning microscope. Females of the calanoid copepod Temora stylifera were fed with FitcDx-encapsulated liposomes alone or mixed with the dinoflagellate alga Prorocentrum minimum. Control copepods were incubated with the P. minimum diet alone. Egg production rates,percentage egg-hatching success and number of faecal pellets produced were evaluated after 24 h and 48 h of feeding. Epifluorescence of copepod gut and faecal pellets indicated that the liposomes were actively ingested by T. stylifera in both experimental food conditions, with or without the dinoflagellate diet. Ingestion rates calculated using 3H-labelled liposomes indicated that females ingested more liposomes when P. minimum was added to the solution (16% vs 7.6% of uptake). When liposomes were supplied together with the algal diet, egg production rate,egg-hatching success and faecal pellet production were as high as those observed for the control diet. By contrary, egg production and hatching success were very low with a diet of liposomes alone and faecal pellet production was similar to that recorded in starved females. This results suggest that liposomes alone did not add any nutritive value to the diet,making them a good candidate as inert carriers to study the nutrient requirements or biological activity of different compounds. In particular,such liposomes are proposed as carriers for diatom-derived polyunsaturated aldehydes, which are known to impair copepod embryo viability. Other potential applications of liposomes as a delivery system of drugs and nutrients in copepod mass cultivation, or as carriers of pollutants to study copepod physiology in ecotoxicological experiments, are also discussed.

The aim of the present study was to evaluate the sustainability and welfare of extensively farmed Podolian cattle. A trained interviewer visited 50 farms and filled in a checklist which included four cards corresponding to the following animal categories: calves, replacements, feeders and adults. The analysis of the farming system showed that animals were able to express their main behavioural patterns. In addition, recorded animal-related variables indicated that Podolian cattle could benefit from high standards of welfare. Sustainability of the Podolian farming system in terms of human edible returns was evaluated for two production systems producing 10-month-old calves (10 month) and 18-month-old young bulls (18 month), respectively. Edible returns for humans were low when all animal intakes were considered for both production systems. However, if returns were computed using not only the amount of food used by the animals but also consumable by humans, yields were much higher for 18-month systems [103% crude protein (CP) and 37.1% gross energy (GE)] and so high that they could not be computed for 10-month systems. These results indicate either a low degree of competition (18-month system) or no competition (10-month system) between humans and Podolian cattle. Perceptions of sustainability and welfare of Podolian cattle may promote a favourable positioning of products in premium-price markets and help preserving this breed and the related traditional farming system.

Twenty weaned female buffalo calves were used to evaluate the effect of space allowance in relation to their body surface area on a range of behavioural and physiological parameters. Body surface area in m2 was calculated as 0.12 body weight0.60. Ten calves received 50% of body surface as space allocation (Group 50), 10 others received 90% of body surface area (Group 90). Animals in Group 50 lay with a lower number of outstretched legs than calves in Group 90. Buffaloes from Group 50 were observed standing more frequently than animals from Group 90 ( P < 0.001). The proportions of idling ( P < 0.01) and lying idle observations ( P < 0.001) were higher for Group 90 than for Group 50. Group 90 performed a higher number of non-agonistic interactions than Group 50 ( P < 0.01), whereas the opposite was observed for the number of agonistic interactions ( P < 0.01). When exposed to open field testing, Group 50 animals displayed an increased duration of movement, number of galloping events and more vocalisation. Neither immune responses to phytohemagglutinin and ovalbumin nor the cortisol response to exogenous ACTH were affected by treatment. It was concluded that 50% of body surface area may be an inadequate space allowance for weaned calves.

The aim of this study was to evaluate the intra-observer repeatability of some animal-related variables which could be used in welfare-assessment protocols at farm level. Recordings were performed on seven dairy farms (four for cattle and three for buffaloes). The animals were observed on three occasions at three-week intervals. The variables collected for each animal were behaviour during milking (stepping and kicking), avoidance distance, lameness, and cleanliness. For each farm and each variable, intra-observer repeatability was computed using the Kendall coefficient of concordance (W). A ratio between the variance of the animal and the sum of the latter with the error variance was also calculated using a model of analysis of variance with one factor (animal), to give a further measure of repeatability. These two methods yielded similar results. In particular, in dairy cattle, a high repeatability for avoidance distance, stepping, lameness and cleanliness was observed, whereas for buffaloes avoidance distance and stepping were the repeatable variables.

Twenty Comisana ewes, with no history of mastitis, were divided into 2 groups of 10 and housed separately in 2 rooms within the same building. Ewes in the machine milked (MM) group were removed from their lambs 48 h postpartum and subjected to machine milking twice daily thereafter. Lamb suckled (LS) ewes were suckled by their lambs throughout the trial. The concentrations of the main microbial groups (mesophilic bacteria, pyogenic streptococci, total coliforms, total staphylococci, yeasts and moulds) were measured weekly from both the air and potential sources of udder contamination such as room surfaces, lambs’ mouths, milking operators and milking equipment. The cell-mediated immune response in vivo of ewes to phytohemagglutinin (PHA) injection was evaluated at days 7, 21 and 35 of the study period. At days 7 and 21, ewes were intravenously injected with chicken egg albumin (OVA). Antibody titers were determined immediately before the first antigen injection and then again at days 21 and 35. On each sampling day, an aliquot of blood was analyzed for white blood cell (WBC) differentials. Individual milk samples were analyzed weekly for somatic cell (SCC) and polymorphonuclear neutrophil leukocyte count (PMNLC), and for bacteriological characteristics and composition. The main carriers of milk contamination were the hands of milking operators and mouth of suckling lambs. TheMMewes displayed a lower (P < 0.05) immune response to PHA than the LS animals at day 21. However, no significant differences were found in WBC differentials and antibody titers to OVA. The MM ewes had higher concentrations of PMNL (P < 0.05) and staphylococci (P < 0.01) in their milk than the LS ewes. Milk yield and protein content were not affected by treatment, whereas the fat content was lower (P < 0.01) in the milk of the LS ewes. Results suggest that early transition to machine milking may impair the responsiveness of the ewe immune system. Maximum care should be taken to control the hygiene of the udder, milking equipment and personnel in order to minimize the risk of intramammary infections.

Female water buffalo ( Bubalus bubalis) calves (n = 28) aged 7 to 10 d were divided into four groups of seven animals each to examine the effects of space allowance (Group A: 2.6 indoor m2 + 2.0 outdoor m2/calf; Group B: 2.6 indoor m2/calf; Group C: 1.5 indoor m2/calf; Group D: 1.0 indoor m2/calf) on behavioral, endocrine, and immune variables for a period of 60 d. Animals were offered 7 L/d of a commercial acidified milk substitute. The calves averaged 45.9 kg initially and 92.4 kg finally. The behavior observations were conducted 7 d after grouping and fortnightly thereafter. At wk 4 and 8, the phytohemagglutinin (PHA) skin test was performed to induce aspecific delayed hypersensitivity. At wk 1 and 3, calves were injected i.m. with keyhole limpet hemocyanin. Antibody titers were determined at weekly intervals for 7 wk. Calves in pens with greater space allowance (Groups A and B) were less active than Groups C and D ( P < .001). The latter groups were also observed feeding more often at wk 7 ( P < .01). Calves provided with an outdoor paddock spent less time standing than Groups C and D ( P < .01), and lay with a greater number of outstretched legs ( P < .001). Groups C and D showed a lower reaction to PHA in both skin tests than did Groups A and B ( P < .001 and P < .05, respectively). Group A showed an antibody response consistently higher than groups B, C, and D ( P < .01, P < .05, and P < .05, respectively). At the end of the experimental period, the calves were subjected to an isolation test lasting 10 min. Group D showed a longer duration of movement with respect to Groups A and B ( P < .01); animals from Group C walked more than did Group A ( P < .05). Cortisol concentration evaluated 0, 10, 45, 90, 150, and 225 min after separation from the group was higher in Groups C and D than in Groups A and B ( P < .01). For all animals, the highest cortisol level was observed immediately after the isolation test ( P < .001). Space restriction resulted in evidence of stress in the animals as shown by alterations in a number of physiological responses. However, the use of small groups of only seven animals per pen may have affected their reactions to space restriction. It is possible that using larger groups could change these conclusions.

This paper describes a goat serum antigen (A2) located on a lipoprotein molecule. This antigen is inherited in a simple Mendelian manner and the gene controlling its synthesis is allelic to the one controlling the synthesis of A1, another lipoprotein determinant. In the heterozygous animal the two determinants reside on distinct molecules. This system, by comprising two codominant alleles, is particularly suitable for the solution of disputed parentage cases and the understanding of lipoprotein biosynthesis and structure.

This study aims to investigate the relationship between human behaviour, buffalo behaviour and prevalence of oxytocin injection at milking. The research was carried out on 17 buffalo farms. On each farm the stockperson's behaviour toward the buffaloes and the buffaloes' behaviour were observed during one afternoon's milking. Stockperson's behaviour was observed from collection of the animals in the waiting area to exit from the milking parlour. The following variables were recorded: number of positive, neutral and negative interactions. The number of steps and kicks were recorded from the entrance to the milking parlour to the removal of cups whenever the stockperson was less than 0.5 m from the animal. The prevalence of oxytocin injection at milking was recorded on two separate occasions with a 5-month interval to obtain a measure of the long-term consistency of this variable. Significant correlations were found between stepping and kicking, stepping and prevalence of oxytocin injection, kicking and prevalence of oxytocin injection and stockperson positive interactions and kicking. These results indicate that stockperson behaviour is related to buffalo behaviour at milking and the latter to the use of oxytocin injections. In addition, prevalence of oxytocin injection proved to be highly reliable when re-tested five months later.

The aim of this study was to evaluate the repeatability of some animal related variables, which could be used in proto- cols developed for assessing animal welfare at farm level. Recordings were performed in seven dairy farms (four for cat- tle and three for buffaloes). The animals were observed on three occasions at three-week intervals. The variables col- lected for each animal were the following: behaviour during milking (stepping and kicking), avoidance distance, lame- ness and cleanliness. For each farm and each variable repeatability was computed using the Kendall coefficient of con- cordance (W). In buffalo farms avoidance distance may be considered highly reliable (W > 0.64), whereas in dairy cat- tle its reliability ranged from medium (W = 0.43 to 0.59) to high (W = 0.64). Behavioural recordings at milking showed that the reliability of stepping was either medium or high for both buffaloes and cattle (W = 0.51 to 0.66 and W = 0.52 to 0.76 for buffaloes and cattle, respectively). Conversely, kicking was less reliable. In cattle farms the reliability for cleanliness ranged from medium (W = 0.51) to high (W = 0.62 to 0.71), whereas, it was not reliable in the sole buffalo farm where this variable was monitored. In cattle farms, the concordance for lameness score was high in two farms (W = 0.62 and 0.66) and moderate in one farm (W = 0.43), whereas no animals displayed lameness in the fourth farm. In all buffalo farms no animals showed lameness. For each species, the Kruskal-Wallis analysis of variance with one factor (farm) was performed to evaluate the effect of farm on recorded variables. For cattle, avoidance distance (P stepping (P nificantly different between farms. In buffaloes a significant effect of farm was observed only for avoidance distance (P that avoidance distance was lower in buffaloes than cattle (P criminate among farms. Lameness and cleanliness scores were able to discriminate only cattle farms, whereas these two parameters, albeit feasible, seem to have low significance for buffaloes. Although stepping during milking was reliable and different among cattle farms, its use in on-farm assessment may be difficult because it is more time consuming, thus less feasible.&nbsp

Sixty-seven multiparous (group M) and 79 primiparous (group P) Friesian cows were used to assess the consistency of the order of entry and the preference for a determinate side into the milking parlour. The Kendall’s coefficient of concordance showed a constancy of the entrance order into milking parlour for both primiparous and multiparous subjects (0.36, P

The present study aims to assess the relationship between consumer perception and on-farm assessment of animal welfare performed using the Animal Needs Index 35L (ANI). Two tie-stall, 2 straw yard and 2 cubicle farms were scored by trained assessors using the ANI and filmed to produce six 3.5 min videos. Each of them contained 4 clips: barn overall view, feeding, milking and individual animals. Ten untrained observers were asked to elicit terms describing how they perceived the observed farming systems to affect animal welfare using Free Choice Profiling (FCP). Data from FCP were subjected to Generalised Procrustes Analysis (GPA). GPA showed a significant consensus among observers. Observers characterised the first dimension with terms ranging from constrictive to comfortable conditions and the second one in terms of cleanliness/dirtiness. Principal Component Analysis (PCA) was conducted using the scores of the farms on the first two dimensions of GPA and the data gathered through the ANI. The first dimension of GPA (Comfortable), ANI's sheet 1 (Locomotion), sheet 2 (Social interaction) and sheet 6 (Total score) showed high loadings on the first component of PCA, whereas the second dimension of GPA (Clean), sheet 3 (Flooring) and sheet 4 (Stockmanship) were correlated with the second component of PCA. We concluded that FCP may be used to elicit lay person perception of welfare-related characteristics of dairy cattle farming systems, thus providing a tool to study the relationships between consumer perception and on-farm assessment of animal welfare.

Twenty-eight buffalo cows were used to evaluate the effect of housing system on a range of behavioural and physiological variables. Fourteen cows were group-housed in a loose open-sided barn with a concrete floor and 10 m2 per head as space allowance (group IS). Fourteen others were group-housed in a similar barn but they could also benefit from an outdoor yard with 500 m2 per head as space allowance, free access to potholes for wallowing and spontaneous vegetation (group TS). Animals were subjected to six sessions of instantaneous scan sampling at 10-day intervals. Behavioural variables were expressed as proportions of subjects observed in each category of posture and activity. Phytohaemagglutinin (PHA) was used to perform a skin test based on non-specific delayed type hypersensitivity, whereas 20 mg of ovalbumin were injected subcutaneously to evaluate humoral immune response. Blood samples for evaluation of cortisol concentration were collected immediately prior to exogenous porcine ACTH injection and 1, 2 and 4 h after. The metabolic status of the animals and milk production were also monitored. The proportion of idling animals was higher in group IS than in group TS (P < 0.001). More IS buffalo cows were observed eating at the manger than TS animals (P < 0.001). A higher proportion of TS animals were observed in the sun (P < 0.001). Grazing and bathing activities were recorded only for TS animals. Our findings suggest that buffalo cows kept in intensive conditions and having no access to ample yards and potholes may extend their periods of idling with negative effects on the state of welfare. Immune responses, metabolite concentrations and milk production were not affected by treatment, whereas cortisol levels were higher in IS animals (P < 0.05). The provision of a housing system similar to natural conditions was able to improve the welfare of buffalo cows as indicated by the expression of some species-specific natural behaviours. Such conditions were also associated with lower adrenal cortex response to ACTH injection, possibly as a consequence of the higher degree of initiative allowed to TS cows.

Given the absence of a ‘Golden Standard’ for the objective determination of welfare, the collection and interpretation of data involving different parameters is essential for assessing the well-being of farm animals. The choice of parameters and the relative weights assigned to each of them are crucial for the outcome of the assessment. Both elements involve a certain degree of subjectivity. In this paper we discuss the basics of different methods used to integrate welfare parameters, focussing on the issue of scientific objectivity. We begin by addressing parameter selection, the assignment of parameter weightings or rankings and the qualifications necessary for ‘experts’ designing and applying the methodology. Five different approaches to integrating parameters are then discussed. The paper does not state a preference for any method, but aims to encourage discussion of key elements involved with the on-farm assessment of welfare.

From April to August 2008, twelve Podolian subjects, aged about 11 months at the beginning of the experimental period, were used to evaluate the effect of rearing system (Confined vs. Freerange) and season (spring vs. summer) on their behaviour and meat quality. Nine sessions of behavioural observations were performed. During a 6-h period, the behaviour of a focal animal, was continuously monitored. In each session a different animal was chosen. All the animals were slaughtered at 18 months of age. Walking (P

Six adult and six juvenile goats were used to estimate their feeding preferences when three grasses (ryegrass, barley and brome) and two legumes (clover and alfalfa) were tested. Each grass was separately tested with both clover and alfalfa. A comparison between alfalfa and clover was also performed. The plants were harvested just before their utilization, chopped into 4- to 6-cm pieces and offered to the animals in plastic baskets. Preferences were evaluated by means of a series of seven short-term trials. Each short-term trial consisted of two single-food presentations and one choice-presentation. In the first single-food presentation animals received one of the two plants to be tested. In the second single-food presentation they were offered the alternative plant species. In the choice presentation they received the two species simultaneously. During each presentation animals were allowed to eat for 20 min or take a maximum of 50 bites. In the single-food presentations bite rate, bite mass and intake rate of both juvenile and adult animals were influenced by the plant species offered. Indeed, all goats tended to display a higher intake rate and bite mass for grasses than for legumes. A grass/legume ratio of the intake rates was also computed for each grass-legume comparison. These ratios tended to be higher for juvenile than for adult goats. The results of the choice presentations indicated that food preferences were not affected by age as all goats preferred grasses to legumes and clover over alfalfa. This experiment showed that age may not play an important role in the development of food preferences.