An observational case study was designed to highlight issues associated with a possible expansion of dairy buffalo (Bubalus bubalis) farming outside the traditional coastal plains of southern Italy. Twenty pregnant buffaloes were transferred to a hilly inland farm. After calving, production and reproduction data were collected monthly throughout lactation. From 4 to 6 months of lactation, buffaloes were enrolled in a feeding trial to evaluate the effects of locally grown forages (maize silage vs. hay) on milk production and in vivo digestibility. Sensory properties of mozzarella cheese produced at a local dairy were also evaluated. No obvious effects of diet were found. Compared to the data recorded in the previous lactation completed in the farm of origin, milk yield was reduced by 37.2%, and milk protein by 6.1%, whereas milk fat improved (+4.5%). A lower pregnancy rate (−13.3%), increased days open (+122%), and a prolonged intercalving period (+26.9%) were also observed. Lactation length was shorter than the standard value of 270 d. The results showed that peculiar reproductive characteristics, lower environmental temperatures, and the specificity of the mozzarella production process are the main problems to be addressed in an expansion of buffalo farming outside traditional areas.

This paper presents the results of a survey carried out in 68 dairies in southern Italy on the manufacturing processes of traditional Italian Caciocavallo cheese varieties. Following a study of the relevant literature, the various cheesemaking processes were analysed and the implications of different cheesemaking procedures were explored. The manufacturing variations able to influence the organoleptic characteristics of Caciocavallo cheese were milk and rennet types, procedures for curd acidification and stretching, salting and ripening conditions, and smoking treatment. This survey is designed to guide producers and consumers alike with respect to the perceivable effects of manufacturing variants on cheese quality.

The incidence and the potential risk factors for cross-sucking in calves and intersucking in lactating cows were assessed in 73 buffalo farms located in the area of PDO ‘Mozzarella di Bufala Campana’. Two trained assessors collected the data through a questionnaire including resource and management measures, which were directly taken and asked to the farmer, respectively. The incidences of cross-sucking, intersucking and mortality were also asked to the farmer. The farm was used as experimental unit. For each discrete risk factor, an ANOVA was performed to assess the association with outcome variables (i.e. the incidence of cross-sucking and intersucking), whereas for continuous risk factors the association was tested using Pearson correlation coefficients. Cross-sucking and intersucking were observed in 91% and 45% of the farms, respectively; their mean incidences were 21.9 ± 22.3 and 1.2 ± 1.9 (mean ± SD), respectively. Cross-sucking was negatively correlated with the number of calves in the collective pen (n = 34, r= −0.47, p=.01). Furthermore, cross-sucking tended to be positively correlated with calf mortality from 24 h to weaning (n = 34, r=.27, p=.11). The incidence of intersucking was positively correlated with the number of buffalo cows on the farm (n = 73, r=.26, p=.03), the number of lactating buffalo in the pen (n = 73, r = 0.23, p=.04), the percentage of weaned calves cross-sucking (n = 73, r=.29, p=.01), the percentage of heifers and pregnant heifers cross-sucking (n = 73, r=.61, p=.001 and n = 73, r=.52, p=.001; respectively). As expected the percentage of buffalo cows with nose-ring was highly correlated with the incidence of intersucking (n = 73, r = 0.75, p=.001). Moreover, the expression of cross-sucking in the categories of weaned calves, heifers and pregnant heifers induced a higher incidence of intersucking (F1,69=5.5, p=.02, F1,70=28.8, p=.0001 and F1,71=15.5, p=.0002; respectively). We conclude that appropriate management strategies should be implemented in order to reduce the incidence of cross-sucking and intersucking. In addition, the onset of cross-sucking in replacement stock, such as calves and heifers, may favour the development of intersucking in their later lives. Acknowledgements The research was funded by University of Basilicata (Intersucking nella bufala Mediterranea italiana: analisi dei potenziali fattori di rischio).

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 basedmeasures (i.e. Welfare Quality (R)). 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 (R) was used for this calculation) was calculated using the Spearman correlation coefficient (r(s)). 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 r(s) = 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 (R) principle "Good health" and the somatic cell counts (r(s) = - 0.500; P = 0.003), the number of lactating animals (r(s) = - 0.360; P = 0.040), the total number of cows per herd (r(s) = - 0.339; P = 0.053), whereas the correlation of the same principle with mortality (r(s) = - 0.315; P = 0.074) and calving interval (r(s) = - 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 chapter will focus on the effects of interaction of river buffaloes with the environment both in confinement and in extensive conditions, on their behavior and welfare. Firstly, the time and place of domestication are described. Sections on time budget, foraging and feeding habit, maternal and social behavior, and some aspects of style of interaction with the environment and temperament are then elucidated. Subsequently, focus is shifted on the quality of relationship between stock-people and animals, which is an important aspect affecting animal welfare in modern farms. Finally, the welfare consequences of intensive farming on buffalo welfare are evaluated using a number of animal-based indicators.

Table S1 Sequence alignment of upstream and downstream regions at position 1022. Capital letters indicate the polypirimidine-reach region. Table S2 Primer sequence of oligonucleotides used in AS-PCR. Table S3 Hardy-Weinberg equilibrium based on g.1022G>A in water buffalo infected and not infected with Mycobacterium bovis. Table S4 Association between resistance to Mycobacterium bovis infection and the g.1022G>A polymorphism in the replication study group 2. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

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<0.01 for groups HL and HH, respectively) and vigilance (2.58±0.46% v. 1.20±0.46%; P<0.05 for groups HL and HH, respectively). Immune responses were not affected by group size.

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.

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.

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; P<0·001), F2G1 (W=0·779; χ2=5046·81; P<0·001) and F2G2 (W=0·624; χ2=2030·48; P<0·001). Spearman rank correlation coefficients indicated that the more productive cows in groups F1 and F2G1 tended to enter the milking parlour first (r s=−0·221 and r s=−0·215; P<0·10; respectively). In group F2G1, a negative correlation was found between duration of milking and order of entry in the milking parlour (r s=−0·265; P<0·05). Animals in group F2G2 (r s=0·334; P<0·05) with higher days in milk entered the milking parlour latterly. In all three groups, 68 cows (45·9%) preferred the right side of the milking parlour, 73 the left side (49·3%) and the remaining seven (4·8%) showed no preference. Finally, negative correlations were found between mean entrance order and parity for both groups of Farm 2 (r s=−0·319; P<0·05 and r s=−0·325; P<0·05 for F2G1 and F2G2, respectively). As buffaloes showed higher entrance order consistency and side preference than other domestic ruminants, it is concluded that management practices that disturb their choice should be avoided in order to minimise stress during farming routines.

The aim of this study was to investigate the effect pre-parturm habituation in the milking parlour on behaviour and lactation performance of buffalo heifers. Sixteen buffalo heifers in late gestation were used for this study. The animals, with an age ranging from 30 to 44 months at the start of the study, were equally allocated into two treatments and balanced for estimated calving date. Eight animals received pre-parturm habituation for 10 days before the estimated calving date (group H), while the eight others received no treatment and served as the control (group C). During the pre-partum habituation sessions, group H animals were moved to the milking parlour and left for 10 min in the milking stall once each day, where the udder was thoroughly washed with warm water, wiped with a disposable towel and massaged. The behaviours, registered from entrance into the milking stall to exit, were step and kick. After calving milk flow profiles, milk yield and milk quality variables were determined along with behavioural recordings. The H animals during the 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. Animals from 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 number of steps (P < 0.001) and kicks (P < 0.001) was observed as lactation proceeded, whereas for group H only a tendency for a reduction over lactation was detected. Pre-partum habituation did not significantly affect milk quality or milk flow variables. Milk yield in the first 3 min of milking (P < 0.001), and average milk flow (P < 0.001), increased throughout the experimental period, whereas the duration of the pre-milking phase decreased as lactation proceeded (P < 0.001). This study shows that buffalo heifers exposed to a pre-partum habituation 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 in buffalo heifers during milking. (C) 2014 Elsevier B.V. All rights reserved.

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.

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 2.5min, 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.

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 10days of age to one of four treatments: A—milk replacer with no additive, B—milk replacer supplemented with 0.17g/L of viable (2×l09cfu/g) E. faecium SF68 culture for 3 consecutive d at 7day intervals for 11wks, C—milk replacer supplemented with E. faecium SF68 daily for 4wks, D—milk replacer supplemented with E. faecium SF68 daily for 11wks. 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 2wks. 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. 4wks), or dispersed over 11wks (i.e. 3 consecutive days at weekly interval), allowed the effect of the probiotic to be achieved at lower cost.

Forty buffalo heifers of approximately 19 months of age were used. Subjects, with a mean live weight of 390 kg, were randomly and equally allocated for 12 weeks to 4 groups. Thus, the experimental design was a 2 x 2 factorial with two space allowances (2.3 indoor slatted floor and 2.3 indoor slatted floor + 15 outdoor yard m2/head) and two dietary protein levels (8.9% and 13.4% of DM). All animals were weighed at group constitution and then at monthly intervals. Behaviour. Three sessions of behavioural observations were made every 10 minutes over a 6 hour period (10.00 to 16.00 h), recording posture (standing or, when lying, number of outstretched legs) and activity such as feeding, ruminating, drinking, locomotion, idling. Rapid behaviours such as agonistic and non agonistic interactions were recorded continuously........

The aim of this study was to evaluate whether housing conditions allowing the animals to lie in the mud and performmore physical exercise can negatively affect reserve mobilization and milk production. In addition, the effect of calvingdistance on blood metabolites was assessed. The experiment was conducted on twenty-eight lactating buffalo cows,equally allocated to two treatments. Fourteen cows were group-housed in a loose open-sided barn with a concrete floorand equipped with self-locking stanchions, where they received 10 m2/head of space allowance, as in intensive systems(Group IS). Fourteen others were group-housed in a similar barn but they also had the benefit of an outdoor yard with500 m2/head as space allowance, including spontaneous vegetation and potholes for bathing and wallowing, as in traditionalsystems (Group TS). Animals were included in the experimental groups 5 days after calving. Daily milk yield, andmilk fat, protein and somatic cell content were determined 4 weeks after grouping (about 35 days after calving) and thenat monthly intervals (5 recordings). Blood samples were collected from the jugular vein in vacuum tubes 10 days aftergrouping (i.e. 15 days after calving) and then at 10 day intervals (17 collections). After centrifugation, plasma and serumaliquots were frozen stored until metabolite determinations (glucose, cholesterol, triglycerides, NEFA, urea, creatinine,albumins, total proteins, calcium, phosphorus, bilirubin, alkaline phosphatase, aspartate aminotransferase, alanineaminotransferase, lactate dehydrogenase and γglutamyl transferase). Treatment did not affect daily milk yield and milkfat, protein or somatic cell count content. Blood metabolite levels were not affected by treatment and interaction treatmentx time. Conversely, as expected, calving distance had an influence on most of these variables (P<0.05÷0.001). Inparticular, in the first two months after parturition, glucose concentration decreased, whereas NEFA and cholesterolincreased as a consequence of the high energetic requirements of animals in the first stage of lactation. The systems thatallow the expression of some basic natural behaviour did not have any detrimental effect on buffalo metabolism and production,thus they should be encouraged.

The present paper addresses the issue on buffalo welfare. Firstly, the biological characteristics and behavioural needs of buffalo are considered. Subsequently, the effects of intensive farming and some animal-related 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 behavior; 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.

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 3 kg/d of CC were replaced by an equal quantity of ExpC. Digestibility of the diets and milk yield 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 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 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 85min 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.