Background-deflection Brillouin microscopy reveals altered biomechanics of intracellular stress granules by ALS protein FUS

Altered cellular biomechanics have been implicated as key photogenic triggers in age-related diseases. An aberrant liquid-to-solid phase transition, observed in in vitro reconstituted droplets of FUS protein, has been recently proposed as a possible pathogenic mechanism for amyotrophic lateral sclerosis (ALS). Whether such transition occurs in cell environments is currently unknown as a consequence of the limited measuring capability of the existing techniques, which are invasive or lack of subcellular resolution. Here we developed a non-contact and label-free imaging method, named background-deflection Brillouin microscopy, to investigate the three-dimensional intracellular biomechanics at a sub-micron resolution. Our method exploits diffraction to achieve an unprecedented 10,000-fold enhancement in the spectral contrast of single-stage spectrometers, enabling, to the best of our knowledge, the first direct biomechanical analysis on intracellular stress granules containing ALS mutant FUS protein in fixed cells. Our findings provide fundamental insights on the critical aggregation step underlying the neurodegenerative ALS disease

Influence of space allowance on the welfare of weaned buffalo (Bubalus Bubalis) calves

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

Sustainability and welfare of Podolian cattle

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

Estimation of the methane emission factor for the Italian Mediterranean buffalo

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

Housing system and welfare of buffalo (Bubalus bubalis) cows

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

The qualitative assessment of responsiveness to environmental challenge in horses and ponies.

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

Use of peas in organic buffalo farming: effects on nutrient digestibility and milk production

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