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Upon in vivo administration of nanoparticles, a protein corona forms on their surface and affects their half-life in circulation, biodistribution properties, and stability; in turn, the composition of the protein corona depends on the physico-chemical properties of the nanoparticles. We have previously observed lipid composition-dependent in vitro and in vivo microRNA delivery from lipid nanoparticles. Here, we carried out an extensive physico-chemical characterisation to understand the role of the lipid composition on the in vivo fate of lipid-based nanoparticles. We used a combination of differential scanning calorimetry (DSC), membrane deformability measurements, isothermal titration calorimetry (ITC), and dynamic light scattering (DLS) to probe the interactions between the nanoparticle surface and bovine serum albumin (BSA) as a model protein. The lipid composition influenced membrane deformability, improved lipid intermixing, and affected the formation of lipid domains while BSA binding to the liposome surface was affected by the PEGylated lipid content and the presence of cholesterol. These findings highlight the importance of the lipid composition on the protein-liposome interaction and provide important insights for the design of lipid-based nanoparticles for drug delivery applications.

Objective: To conduct a bibliographic review of the inventory and distribution of the buffalo herd, leading dairy buffalo breeds, and to map the main research topics for dairy buffaloes, emphasizing feeding methods and their effects on milk and mozzarella cheese quality. Design/Methodology/Approach: Analysis of the main research topics on dairy buffalo, through a wide review of specialized journals. Results: The production and processing of buffalo milk has gained relevance in recent years along different latitudes thanks to its nutritional qualities and the international regard for products such as mozzarella cheese. The main studies are carried out in Asia, Italy, and Brazil, emphasizing that diets are a determining factor in yield and quality of milk and its derivatives, but that genetics, environment, and animal management are what in the end model these characteristics. Study Limitations/Implications: To conduct further research on dairy buffalo, especially in Mexico, where it has important development opportunities. Findings/Conclusions: The bibliographic body of work presents practical restrictions, advances are recognized, and also the need to further research topics such as reproduction and animal welfare, management and valuation of buffalo milk and its derivatives, with the opportunity to explore organic production.

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. Crosssucking 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.

ClassyFarm is an integrated system aiming at the categorization of the farms according to a risk assessment methodology (RA). It is an Italian innovation that allows to facilitate and improve the collaboration and the dialogue between the breeders and the competent authority, in order to raise the level of safety and quality of food of animal origin. ClassyFarm collects, gather and process data, referred to the following evaluation areas: biosecurity, animal welfare, health and production parameters, animal nutrition, antimicrobial usage, injuries detected at slaughterhouse. ClassyFarm can be applied to several livestock species, included buffalo farms. On the request of the Italian Ministry of Health, CreNbuf in collaboration with CReNBa, have develop a checklist (CL) for the animal welfare assessment in buffalo species, based on RA and included in the ClassyFarm system. The multiple-choice CL consists of 80 Items. Each Item is scored according to 3 choice categories: “insufficient”, “acceptable” and “excellent”. The assessment system includes non-animal based (N-ABMs) and animal-based measures (ABMs). N-ABMs are divided into 2 macro-areas: Area A (28 Items) “Management factors”; Area B (30 Items) “Housing factors”. ABMs are assessed in Area C (14 Items). The CL has been tested in 341 farms, with an average of 353 head (min10, max 2240). The average overall welfare value was 61.27% (on a scale of 0 to 100%). The values of the specific areas were: Area A 60.20%; Area B 41.34%; Area C 71.55%. At least one potential legislative non-compliance was recorded in 39.8% of the farms. These CL represent a functional and smart instrument to assign an animal welfare index to each farm, and to provide to veterinarians and breeders a tool for improving farm management and housing conditions by using the data collected in each Area, while respecting farm sustainability, giving answers to consumers and add value to farmers’ correct activities. The Italian Ministry of Health is promoting this system, unique in its kind, also in the European and international context, raising a wide and strong interest.

An expert opinion elicitation, based on a modified Delphi technique, was organized to collect the opinion of 14 Italian veterinarians with the aim of conducting a hazard and a welfare promoter characterization for defining and weighing a list of management and housing factors potentially associated with negative or positive welfare outcomes in buffalo kept in loose housing systems. In addition, the 14 experts judged a set of animal-based measures (ABMs), rating their appropriateness and the level of animal pain and suffering due to the welfare consequences they measure. Experts were asked to score 52 hazards, 45 welfare promoters and 14 ABMs. Management and housing hazards, that were determined to be associated with a very high negative impact on the welfare of buffalo, were: lack of water availability, uncorrected animal grouping strategy, dirty and unmanaged lying area, incorrect milking routine, poor maintenance of the milking system, lack of litter for calves, insufficient shaded lying area and/or lack of any cooling system and presence of inadequate or slippery floor in walking areas. Management and housing welfare promoters dealing with optimal buffalo comfort around resting obtained the highest ratings, in particular: housing free for all animals and the possibility for adult buffaloes to access a loafing area of at least 7 m²and/or to a pasture for at least 60 days a year; cleanleness of the calf boxes, a hospital pen of at least 8 m²/parturient animal. In addition, high scores were given to appropriate grouping, adequate temperature, humidity and ventilation, presence of experienced and trained stockpersons. As to ABMs, the highest rankings were given to mortality of calves and adult buffaloes, percentage of animals showing vaginal and/or uterine prolapses, body condition scoring and percentage of deformed claws in adult buffaloes. Our results represent the starting point for the development of the first protocol for the assessment of the welfare of buffalo farmed in loose housing systems.