Laboratory methods (LAB) for feed analysis are reliable but time-consuming and expensive. Near-infrared reflectance spectroscopy (NIRS) offers a non-destructive alternative, and portable devices have increased interest for on-farm real-time monitoring of feed composition. However, their accuracy relative to LAB remains insufficiently validated. This study evaluated two portable NIRS instruments compared with LAB methods for chemical characterisation of total mixed rations (TMR) for dairy buffaloes. 230 TMR samples were collected, scanned with both devices, and analysed with LAB methods. Parameters assessed included dry matter (DM), ether extract (EE), ASH, crude protein (CP), neutral detergent fibre, acid detergent fibre, acid detergent lignin, and starch. Agreement among methods was examined using Bland-Altman and Passing-Bablok analyses, while differences were evaluated with Friedman ANOVA. Principal Component Analysis (PCA) was applied to explore the multidimensional structure of the dataset. Finally, Passing-Bablok regression was used to correct NIRS bias and reassess agreement with the reference method. Good agreement was observed for DM (mean differences 3.12% for NIRS_A, 0.99% for NIRS_B) and moderate for EE (<0.5%), while larger discrepancies occurred for fibre fractions, CP, ASH, and starch. Passing-Bablok regression revealed significant constant and proportional biases, particularly for fibre and starch. Bias correction improved agreement, with DM and EE predictions closely matching LAB values (e.g. DM: slope = 1.00, intercept = 0.95). The PCA indicated that NIRS devices capture overall compositional patterns, with NIRS_B showing lower variability. In conclusion, portable NIRS devices cannot yet replace laboratory analyses for buffalo TMR; however, after bias correction, they may have potential as tools for on-farm monitoring.

IntroductionThe gastrointestinal microbiota of ruminants plays a crucial role in health, influencing immune responses, nutrient metabolism, and environmental impact. While the ruminal microbiota has been widely investigated, the hindgut microbiota, particularly the fecal microbiota, remains less explored. Diet strongly shapes microbial communities, thereby affecting digestion, metabolic pathways, and methane emissions. Next-generation sequencing enables detailed microbiota profiling; however, no studies have characterized the fecal microbiota of Italian Mediterranean buffaloes in relation to diet. This study aimed to evaluate the bacterial composition and dietary influences on the fecal microbiota of dairy buffaloes.MethodsOver 6 months, monthly pooled fecal samples were collected from ~10 to 15% of lactating buffaloes across 10 farms. Concurrently, dietary data were recorded, and total mixed ration samples were analyzed for physicochemical properties and fatty acid profiles. DNA was extracted using the Quick-DNA™ kit, followed by 16S rRNA sequencing on an Illumina MiSeq System. Statistical analyses in R included alpha and beta diversity, differential abundance testing, and one-way ANOVA (p < 0.05; trends at p < 0.10).ResultsIn total, 10 phyla, 13 classes, 26 orders, 47 families, 86 genera, and 120 species were identified. Firmicutes was the most abundant phylum (55.8 ± 3.6%), followed by Bacteroidota (37.7 ± 3.4%). Among dietary variables, the forage-to-concentrate (FC) ratio and linseed (LS) inclusion exerted the greatest influence. Notably, the FC ratio affected beta diversity (community structure) but not alpha diversity (within-sample diversity), whereas LS inclusion influenced both alpha and beta diversity. A low FC ratio promoted phyla negatively associated with fiber digestibility, particularly families Lachnospiraceae and Succinivibrionaceae, consistent with cattle studies linking these taxa to high-concentrate diets rich in fine particles (<4 mm). Linseed inclusion reduced species richness and increased Firmicutes, Spirochaetota, and Proteobacteria, the latter including potential pathogens implicated in ruminal dysbiosis. Conversely, LS inclusion decreased Verrucomicrobiota, a phylum important for gut health and mucus layer maintenance.ConclusionThis study provides the first characterization of the fecal microbiota of Italian Mediterranean dairy buffaloes and highlights its responsiveness to diet. Findings underscore the potential of fecal microbiota as a non-invasive biomarker for evaluating dietary effects, with implications for animal health, productivity, and environmental sustainability.

This study examined the effects of seasonal climatic variations and parity on the milk yield and composition of Holstein (HST), Brown Swiss (BSW), and Montbéliard (MBM) dairy cows in North Tunisia from 2020 to 2024. Test day records (TD) from 199762 observations for different classes of parities (1, 2, 3 and 4+), with calculation of the Temperature-Humidity Index (THI) through Ambient Temperature (AT, ℃) and Relative Humidity (RH, %), were merged with meteorological data for analysis of the effect of seasonal heat stress and parity on the milk yield and composition. Meteorological data indicated a significant seasonal variation, with THI surpassing heat-stress thresholds during summer (mean = 75.29) and reaching its maximum in July and August. According to the results, Holstein cows had the highest peak milk yields (about 24.5 kg/d), but they were also the most sensitive to heat stress, as shown by their significant production decrease in summer. On the other hand, despite environmental challenges, BSW and MBM showed better thermotolerance and maintained consistent lactation. Analysis of milk composition revealed a strong seasonal concentration effect in all breeds, with the percentages of milk fat (MF) and milk proteins (MP) increasing in the summer and autumn months, with BSW producing the highest solid concentrations (up to 3.87% fat). Statistical analysis demonstrated significant interactions among breed, season, and parity (p < 0.05) for all traits except parity with milk fat. The results indicate that, whereas Holsteins maximize performance in temperate environments, incorporating Brown Swiss and Montbéliard genotypes provides a more resilient strategy for maintaining dairy productivity and milk quality in the increasingly hot Mediterranean climate of North Tunisia.

This study examined the impact of heat stress on physiological responses, milk yield and composition, and heat shock biomarkers in 45 dairy cows (Holstein (HO, n = 15), Montbéliard (MT, n = 15), and Brown Swiss (BS, n = 15)) at the same lactation stage and reared under uniform management and climatic conditions in Jendouba, North Tunisia. During a three-week summer period (July - August 2024), cows were exposed to high temperatures and humidity with THI levels ranging from 70.11 to 80.60. Significant breed-specific differences (P < 0.05) were observed in rectal temperature, respiration rate, heart rate, and panting score, with HO cows showing the highest stress indicators. BS cows spent more time lying (69.99%) than MT (48.88%) and HO (39.99%). Although HO had the highest average daily milk yield (25.97 ± 5 L/day) across the study period, production declined significantly from week 1 to 3 by 11.58%, while BS maintained stable output (P > 0.05) for a decrease of 2.79%. Milk composition also varied, with HO showing reduced fat and protein levels under stress, whereas BS remained more stable. Plasma cortisol levels increased across all breeds by week 2 (up to 3.84 ng/ml), but HO had the lowest HSP70 concentrations (4.64 - 6.53 ng/ml), indicating lower thermotolerance. Overall, BS cows demonstrated greater resilience to heat stress, maintaining physiological stability and milk performance, underscoring the importance of breed choice for climate-adaptive dairy farming.

Recent studies focus on animal welfare, heat stress (HS), climate change, and enteric methane (CH4) emissions from livestock. To mitigate HS effects and reduce CH4 emissions, various strategies exist. This study evaluated a commercial nutritional supplement in lactating dairy buffaloes during summer, assessing its impact on diet digestibility, milk production, and CH4 emissions. The supplement contained yeast, capsaicin, Yucca schidigera, garlic, and other compounds. Fifty-seven buffaloes were divided into three groups: control (C) with a basal diet, and two groups receiving 100 g (T1) or 200 g (T2) of the supplement. TMR, faeces, and milk samples were collected, CH4 emissions were measured using a laser gun-type detector, and animal behaviour was monitored via ear-tag sensors. Results indicate that 100 g/head/d of nutritional supplement improved milk yield (p < 0.01, MY = 8.51 L/head/d) and quality, especially the clotting parameter (p < 0.01, RCT = 19.02 min, a30 = 28.99 mm), diet digestibility (p < 0.01) for dry matter (69.12%), crude protein (67.23%), aNDF (62.39%), and starch (97.38%), and animal welfare compared with the control group (MY = 7.92 L/head/d, RCT = 20.01 min, a30 = 28.23 mm, and 60.47%, 57.80%, 50.87% and 96.31% for DM, CP, aNDF, and starch digestibility) and may help animals to counteract the deleterious effects of HS. Moreover, the nutritional supplement decreases (p < 0.05) CH4 emissions (376.28 and 404.68 g/head/d for T1 and C, respectively). Higher doses (i.e., 200 g/head/d) had significant deleterious effects such as worse digestibility, lower milk quality compared lower doses, and did not reduce CH4 as T1.

The increase in ambient temperature is responsible for a behavioral, physiological, and metabolic responses known as heat stress, which affects dairy cows' general well-being, health, reproduction, and productivity. Focusing on the functioning of the mammary gland, attention has been recently paid to a new method of cell-cell communication mediated by extracellular vesicles, which with their cargo can affect the target cells' phenotypic traits, behavior, and biological functions. This study investigated whether the small extracellular vesicles (sEV) isolated from milk of heat-stressed Holstein Friesian (H) and Brown Swiss (B) cows affect the cellular response of a bovine mammary epithelial cell line (BME-UV1). To this purpose, 8 mid lactation cows, 4 of each breed fed the same diet and kept in the same barn, which experienced the same hyperthermia during a natural heat wave, were chosen to collect 2 milk different samples: under thermoneutrality (TN, d1) and under heat stress (HS, d 8) conditions. The sEV were isolated from skim milk samples through differential centrifugations, characterized for size and concentration by nanoparticle presence of EV markers through western blotting. Then BME-UV1 cells were incubated for 24 h with different cows. In vitro results of BME-UV 1 cells treated with milk sEV H-HS and B-HS showed an alteration of the cell viability and metabolic activity, by reducing or increasing reactive oxygen species accumulation, and suppressing or increasing the expression of stress-associated genes thereby modulating the response of BME-UV 1 according to the animals' thermal condition and the breed. These findings indicated that the small vesicles of Brown milk triggered cellular defense against heat stress, supporting the Brown Swiss breed's thermotolerance.

Precision Livestock Farming (PLF) represents a significant evolution in the livestock sector, promising to transform farms management improving production efficiency and sustainability, products quality, working conditions and animal welfare. While PLF is increasingly utilised under controlled condition typically linked to intensive livestock systems, its implementation in extensive livestock farms - where animals are primarily raised outdoor - is a challenge. Extensive systems, which cover approximately 67% of global agricultural land, have significant socio-economic importance worldwide, both in terms of food supply and the provision and maintenance of ecosystem services. At the farm level, Precision Livestock Extensive Farming (PLEF) - including the use of wearable sensors, environmental monitoring equipment, and remote sensing - can be widely employed in production monitoring, solving management problems, addressing logistical challenges and improving efficiency in resource management and finally in decision-making processes. Through text mining of 710 scientific articles published between 1980 and September 2024, this review identifies key trends, technologies, and gaps in current research and management approaches. Major findings highlight the prevalence of sensor technologies for monitoring animal behaviour and pasture quality, principally from Australia and United States. The review underscores the potential of PLF to enhance sustainable production in extensive systems, but also calls for more research on the integration of advanced data analytics and remote sensing technologies, including edge computing. The findings aim to guide future research and practical implementations, fostering the development of sustainable livestock farming practices globally with a view to multispecies approaches tacking into account also wildlife-livestock interactions.HIGHLIGHTS PLEF shifts livestock system management from manual to (semi-) automated. Text mining shows the evolution and spread of PLEF literature over the years. PLEF tools help address environmental and livestock system challenges.

This study investigated the pattern of 6 behavioral parameters in Holstein dairy cows under heat stress (HS) conditions using a generalized additive mixed model the effectiveness of a commercial electrolyte, osmolyte, and antioxidant blend in mitigating HS-induced adverse effects. The trial was conducted during a severe Italian summer on 84 multiparous dairy cows in the Central Italy, divided into 4 balanced groups (2 treated and 2 control) of 21 cows each. All animals received the same diet, and the treatment groups were supplemented with 3,150 g/d of the additive. Behavioral parameters, namely (HA), rumination time (RUM), eating time (ET), and heavy breathing (OH), were monitored using neck collar sensors. Generalized additive mixed models were used to analyze trajectory changes of these parameters over time and climatic conditions. Results indicated significant variations in only one activity parameter, with treated cows showing increased MA with rising temperature-huHS relief. Rumination time decreased with increasing HS levels but increased also with milk yield and lactaon behavioral parameters.

Background: Cortisol is known as the main hormone released during stress responses in cattle and has been used to assess various stressors, including heat stress. This study investigated hair cortisol concentration (HCC) in different hair coat colors in dairy cows under natural heat stress conditions (temperature humidity index = 75). Methods: Hair samples were collected from the forehead region of ten multiparous cows (Brown Swiss, Montbéliarde, and Holstein) per group color at both the beginning and end of a three-week peak summer period in 2024 in the region of Jendouba, North Tunisia. Cows were grouped according to hair coat color (black, brown, red, white, and yellow) for subsequent analysis. Hair samples were prepared using a methanol-based separation protocol and analyzed via enzyme-linked immunosorbent assay (ELISA). Results: Meteorological data confirmed that cows were sustained under heat stress, with an average temperature humidity index value of 75; results indicated that black hair had considerably more HCC than white hair (p < 0.05). The results showed that there is a significant difference between HCC under three clusters (p < 0.05) according to hair color. Conclusions: The study emphasizes that hair color, along with factors such as breed and environmental conditions, should be carefully considered when using HCC to assess stress in cattle beyond simply black or white hair color.

Near-infrared spectroscopy (NIRS) is an efficient, non-destructive method for evaluating the chemical composition of various compounds. This study aimed to assess both the proximate composition, fibres, and fatty acid (FA) content of Total Mixed Rations (TMRs) in dairy buffalo nutrition. A total of 240 TMR samples were collected from ten dairy buffalo farms across four seasons to develop predictive models using Partial Least Squares Regression (PLSR). Calibration models for dry matter (DM), crude protein (CP), ether extract (EE), and starch demonstrated good predictive accuracy, with coefficients of determination in cross-validation (R2cv) around 0.90 and Residual Predictive Deviation (RPDcv) values exceeding 3.0. Fatty acid models showed slightly lower R2cv values, ranging from 0.80 to 0.90. A good predictive performance was observed for linoleic acid (18:2 n-6) and α-linolenic acid (18:3 n-3), with RPDp values above 3.0, indicating reliable predictions. The inclusion of omega-3-rich compounds in the diet provides significant benefits for both animal health and product quality, highlighting the importance of ration monitoring. The findings confirm that while NIRS is effective for assessing chemical composition, further refinement is needed to improve FA prediction accuracy. These results support the use of NIRS as a practical tool for nutritional monitoring in lactating buffaloes.

Heat stress is a significant challenge in dairy cattle herds, affecting milk production and quality, and generating important changes at the cellular level. Most in vitro research on heat shock (HS) effects on dairy cow mammary cells was focused on medium-long-term effects. In recent years, Fourier transform-infrared (FT-IR) micro-spectroscopy has been increasingly used to study the effects of several external stresses on different cell lines, down to the level of single cellular components, such as DNA/RNA, lipids, and proteins. In this study, the possible changes at the biochemical and molecular level induced by acute (30 min-2 h) HS in bovine mammary epithelial (BME-UV1) cells were investigated. The cells were exposed to different temperatures, thermoneutral (TN, 37 °C) and HS (42 °C), and FT-IR spectra were acquired to analyse the effects of HS on biochemical characteristics of BME-UV1 cellular components (proteins, lipids, and DNA/RNA). Moreover, cell viability assay, reactive oxygen species production, and mRNA expression of heat shock proteins (HSPA1A, HSP90AA1, GRP78, GRP94) and antioxidant genes (SOD1, SOD2) by RT-qPCR were also analysed. The FT-IR results showed a change already at 30 min of HS exposure, in the content of long-chain fatty acids, which probably acted as a response to a modification of membrane fluidity in HS cells compared with TN cells. After 2 h of HS exposure, modification of DNA/RNA activity and accumulation of aggregated proteins was highlighted in HS cells. The gene expression analyses showed the overexpression of HSPA1A and HSP90AA1 starting from 30 min up to 2 h in HS cells compared with TN cells. At 2 h of HS exposure, also the overexpression of GRP94 was observed in HS cells. Acute HS did not affect cell viability, reactive oxygen species level, and SOD1 and SOD2 gene expression of BME-UV1 cells. According to the results obtained, cells initiate early defence mechanisms in case of acute HS and probably this efficient response capacity may be decisive for tolerance to heat stress of dairy cattle.

Recently, much interest has been raised for the characterization of signaling molecules carried by extracellular vesicles (EVs), which are particularly enriched in milk (mEVs). Such interest is linked to the capability of EVs to cross biological barriers, resist acidification in the gastric environment, and exert modulation of the immune system, mainly through their microRNA (miRNA) content. We characterized the small-RNA cargo of colostrum EVs (colosEVs) and mEVs from Italian Mediterranean buffalo through next generation sequencing. Colostrum (first milking after birth) and milk (day 50 of lactation) were sampled from seven subjects from five farms. ColosEVs and mEVs were subjected to morphological characterization, followed by high-depth sequencing of small RNA libraries produced from total RNA. The main difference was the amount of EV in the two samples, with colostrum showing 10 to 100-fold higher content than milk. For both matrices, miRNA was the most abundant RNA species (95% for colosEVs and 96% for mEVs) and three lists were identified: colosEV-specific, mEV-specific and shared most expressed. Gene ontology (GO) enrichment analysis on miRNA targets highlighted many terms related to the epigenetic, transcriptional and translational regulations across the three lists, with a higher number of enriched terms for colosEV-specific miRNAs. Terms specific to colosEVs were related to "cell differentiation" and "microvillus assembly", while for mEV "cardiac and blood vessel development" and "mitochondria" emergerd. Immune modulation terms were found for both sample-specific miRNAs. Overall, both matrices carry a similar molecular message in terms of biological processes potentially modulated into receiving cells, but there is significant difference in the abundance, with colostrum containing much more EVs than milk. Moreover, colosEVs carry molecules involved in signal transduction, cell cycle and immune response, as for mEVs and EVs of other previously characterized species, but with a special enrichment for miRNAs with epigenetic regulation capacities. These beneficial characteristics of colosEVs and mEVs are essential for the calf and could also be exploited for the therapeutic purposes in humans, although further studies are necessary to measure the sanitization treatment impact on EV conservation, especially in buffalo where milk is consumed almost exclusively after processing.

Buffalo breeding in Italy is becoming increasingly economically attractive, with milk production in the Lazio region growing by 15% since 2004. However, the optimal physical characteristics of total mixed rations (TMR) have not been thoroughly investigated. This lack of research suggests the need for greater attention to the physical characteristics of diet, which, combined with its chemical properties, may negatively impact milk yield (MY) and milk quality (MQ), as seen in dairy cows. Therefore, the objective of this study was to assess the effect of diet composition, with a particular focus on the physical component, on MY and MQ, and apparent digestibility (AD) of dairy buffaloes. The study was conducted on 10 dairy buffalo farms in Amaseno valley (Italy). Monthly samples of TMR, faeces, and bulk milk were collected over a 12-month period, alongside interviews with farmers to gather information on MY and diet composition. The results revealed a high variability in the physical-chemical composition of diets across farms, attributable to differences in feeding management practices. Both chemical and physical component were found to be associated with MY, MQ and AD changes. A ration poor in roughage and rich in concentrate negatively affected MQ showing lower milk fat, and higher somatic cells count and milk urea, while a greater presence of coarse particles and high fibre were associated with lower MY. Nevertheless, coarse particles and aNDF were positively associated with milk clotting parameters and aNDF digestibility. Our study emphasised the high importance of fibre for improving MQ of dairy buffaloes. Particle size distribution of dairy buffalo diet is affected by raw material and feeding management.Dairy buffalo unbalanced diets (rich in fibre and low in starch) can negatively affect milk yield.Physical characteristics of diets can positively affect milk clotting parameters of buffalo milk.

The development of detectors for protons and heavy particles is a long-lasting research topic not only for fundamental applications but also, more recently, for monitoring energy and flow of particles in ion beam applications. Nowadays, one of the most demanding application of ion beams is hadron therapy of cancer. In this field, ion beams, mostly proton beams, are used for the controlled treatment of cancer by focusing them onto small volumes to deliver energy to the tumor. The effectiveness and safety of the treatment is enhanced by tuning the beam both in intensity and in position to irradiate the tumor in a controlled way, preserving the surrounding healthy tissues. For this reason, there is an increasing demand of systems optimized for the accurate recording and mapping of the dose delivered during a treatment plan in-situ and in realtime. Here, we present the results achieved by a fully-organic indirect detector in which a flexible Organic Photo-Transistor based on DNTT has been coupled with a plastic scintillator based on polysiloxane. The device has been firstly characterized under 5 MeV proton beams provided by LABEC ion beam center (INFN - FI, Italy) to reproduce the energy range that scattered protons deliver on healthy tissues surrounding the tumor during treatments for prostate cancer. The detector exhibits excellent detecting performances even bent down to a curvature radius of 0.5 mm, at low operation voltages (V = -1 V), reaching a limit of detection of 0.0 6 Gy min -1 . Such results confirm its reliability as a personal dosimeter with high comfort and low risk for the user. Finally we tested the detectors in real clinical conditions employing an anthropomorphic phantom and a therapeutic proton beam (energy range [70–00] MeV) provided by TIFPA Proton Therapy Center (TN, Italy).

In this in vitro study, for the first time was evaluated the antioxidant and anti-inflammatory effect of an Oleuropein-enriched extract (OleE) on bovine mammary epithelial cell line (BME-UV1). OleE was obtained from olives leaves and characterized by HPLC and NMR analysis. Cell viability test indicated that OleE at concentrations of 7.8 up to 250 μg/mL did not exert cytotoxic effect. At concentration of 31.2 up to 250 μg/mL, a dose dependent reduction of ROS production induced by hydrogen peroxide was observed. In addition, OleE at 62.5, 125 and 250 μg/mL showed a dose-dependent reduction in gene expression of TNF, IL1B, and IL10 after exposure to LPS. The downregulation of ROS production and cytokines expression in BME-UV1 by OleE confirmed the antioxidant and anti-inflammatory properties. In vivo experiments will be necessary for future applications of OleE as natural feed supplement in dairy cattle to reduce incidence of oxidative stress in peripartal period.

Increases in temperature and the greater incidence of extreme events are the consequences of the climate change that is taking place on planet Earth. High temperatures create severe discomfort to animal farms as they are unable to efficiently dissipate their body heat, and for this, they implement mechanisms to reduce the production of endogenous heat (reducing feed intake and production). In tropical and subtropical countries, where buffalo breeding is more widespread, there are strong negative consequences of heat stress (HS) on the production and quality of milk, reproduction, and health. The increase in ambient temperature is also affecting temperate countries in which buffalo farms are starting to highlight problems due to HS. To counteract HS, it is possible to improve buffalo thermotolerance by using a genetic approach, but even if it is essential, it is a long process. Two other mitigation approaches are nutritional strategies, such as the use of vitamins, minerals, and antioxidants and cooling strategies such as shade, fans, sprinklers, and pools. Among the cooling systems that have been evaluated, wallowing or a combination of fans and sprinklers, when wallowing is not available, are good strategies, even if wallowing was the best because it improved the production and reproduction performance and the level of general well-being of the animals.

COPYRIGHT © 2023 Basiricò, Abeni and De Palo. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. TYPE Editorial PUBLISHED 13 June 2023 DOI 10.3389/fanim.2023.1221756

The next years will see the completion of the radioactive ion beam facility SPES (Selective Production of Exotic Species) and the upgrade of the accelerators complex at Istituto Nazionale di Fisica Nucleare – Legnaro National Laboratories (LNL) opening up new possibilities in the fields of nuclear structure, nuclear dynamics, nuclear astrophysics, and applications. The nuclear physics community has organised a workshop to discuss the new physics opportunities that will be possible in the near future by employing state-of-the-art detection systems. A detailed discussion of the outcome from the workshop is presented in this report.