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Cortisol is the primary biomarker associated with the hypothalamic–pituitary–adrenal (HPA) axis. This research aimed at assessing the predictive potential and the relationship of cortisol concentrations in four media (blood, milk, whey and hair) with parity, lactation stage and productive classes. Multiparous (n = 30) and primiparous (n = 38) Italian Mediterranean buffaloes were used and assigned to four productive classes (based on percentiles of mature equivalent milk yield (EMY) and mature equivalent energy corrected milk (eECM)), and cortisol concentrations were measured using a in house radioimmunoassay (RIA) method. Parity did not show a significant effect on cortisol concentrations of the four media. The catabolic stage of lactation (up to 90 d in milk (DIM)) was characterised by higher cortisol concentrations compared to the second anabolic stage (beyond 150 DIM) in milk formulations. The plasmatic concentrations of cortisol were higher at the catabolic and the first anabolic stage (91–150 DIM) compared to the second anabolic (p = 0.022 and p = 0.009, respectively). Buffaloes beyond 150 DIM differed from those below 90 DIM (p < 0.001) and between 91 and 150 DIM (p < 0.05) in hair. Finally, hair cortisol concentrations were negatively correlated to EMY, mature equivalent protein content (EPC) (p < 0.01) and eECM (p < 0.05). The whey showed a potential to predict the concentrations of cortisol in whole extracted milk (R2 = 0.31). Hence the development of a real-time automated cortisol measurement in milk could be a valuable tool in Italian Mediterranean Buffalo farming. HIGHLIGHTS: The lactation stage significantly influences plasma, hair and milk cortisol concentrations. Hair cortisol concentrations are negatively correlated to milk quality parameters. The whey is a potential predictor of the cortisol concentrations in whole extracted milk. [ABSTRACT FROM AUTHOR]

Buffalo is a rustic animal; it can adapt and survive in various environments with different topography, climate and vegetation, and has a greater longevity than cattle. The high resistance to parasites, as well as the ability to improve the efficiency of feed consumption in case of forage shortage, means that even in marshy territories, where any other type of livestock or agricultural activity is impossible, buffalo farming can develop an economy. This review aims to present the key research topics, the evolution and geographic distribution of literature on buffalo welfare, and to underscore potentiality and existing knowledge gaps. A comprehensive search on the Scopus bibliometric database was carried out using various welfare-related keywords such as: 'buffalo welfare', 'buffalo calf welfare' and 'buffalo heifers' welfare'. The research identified 308 papers published in English from January 1992 to December 2023 that, after filtering, became 148 eligible records. Descriptive statistics revealed a significant exponential increase in studies on buffalo welfare since 2020, particularly in Italy, followed by Brazil. Text mining and topic analysis revealed that the most frequently mentioned terms were 'milk', 'water', 'behaviour', 'farm' and 'stun', confirming the role and the importance of this species in dairy production. Nine identified topics span diverse subject areas such as 'slaughter process', 'dairy production', 'general welfare', 'housing system', 'heat stress', 'reproduction performance', 'health status' 'breeding management', and 'stress evaluation'. Although academia shows an increasing interest in buffalo behaviour and welfare, continued research is necessary to fully comprehend how to ensure the welfare of this important species. HIGHLIGHTS: Overview of buffalo welfare in scientific literature Usage of new techniques as text mining and topic analysis for the review process Slaughter process and Dairy production were the most frequent topics [ABSTRACT FROM AUTHOR]

The aim of the present study was to determine the effect of including green forage in the diet of Italian Mediterranean dairy buffaloes on the ruminal microbiota, CAZymes profile, functional biomolecules and total antioxidant activity in bulk milk. Sixteen buffaloes were randomly assigned according to lactation number and daily milk production to two homogeneous groups, and for 60 days received each: Group 1, a standard total mixed ration (TMR) or group 2, TMR + ryegrass green forage (30% of diet). The diets of the two groups were iso-nitrogenous and iso-energetic and differed only in the proportion of green forage. Buffaloes that received TMR + green feed had a higher (p <.01) representation of bacteria belonging to the orders Veillonellales, Selenomonadales and Bradymonadales compared with buffaloes that received TMR. The former buffaloes also had a greater (p <.01) abundance of CAZymes of the GT class (GHT4, GT14, GT20, GT26, GT39) and AA class (AA1, AA3, AA6). The milk of buffaloes that received TMR + green feed had a higher (p <.01) antioxidant capacity and greater (p <.01) amounts of the functional biomolecules l-carnitine, propionyl-l-carnitine, acetyl-l-carnitine and δ-valerobetaine. The findings have provided evidence for metabolic and biosynthetic pathways that link green forage with rumen bacteria, CAZymes and the synthesis of amino acids and functional biomolecule in buffaloes. Highlights: Green feed diet in dairy buffaloes favours ruminal microbiota that produces CAZymes. CAZymes support the synthesis of amino acids and functional biomolecules in milk. Functional milk from animals fed natural diets will be more appreciated by consumers. [ABSTRACT FROM AUTHOR]

The aim of this work was to study the effects of red orange and lemon extract (RLE) inclusion on volatile organic compounds (VOCs) and sensory profile of lamb meat aged seven days. A total of 44 Merino lambs were randomly divided into two experimental groups: one that received the RLE (n = 22) and the control group (CON; n = 22). The RLE extract was orally and individually administered (90 mg/kg of live weight) to each lamb every day from colostrum assumption until slaughter (40 ± 1 days). Longissimus lumborum muscle was sampled and dry aged at 4 °C for seven days. The VOC and sensorial analyses were carried out at 1, 3 and 7 days. A total of 65 VOCs were detected. Aldehyde family was the most abundant, followed by ketones and alcohols in both groups. Extending ageing period, content of aldehydes and thiols increased only in control group (p < 0.01), but no differences were observed 24 h after slaughtering (p > 0.05). Regardless of dietary treatment, sensory evaluation, tenderness and juiciness showed a similar trend in both groups. Along ageing days, a significant improvement in these patterns was observed (p < 0.05). Our results showed that RLE inclusion did not have effects on both volatile compounds and sensory profile after slaughtering; ageing improves some sensory characteristics of lamb meat and the inclusion of RLE also seems to have a positive effect on the production of VOCs and on sensory properties such as meaty odour and overall liking. Highlights: Anthocyanins extracted from red orange and lemon extract have antioxidant properties. Red orange and lemon extract is reported to exert antioxidant effect as feed additive in ruminants. Anthocyanins affect volatile compounds (VOCs) of meat only after some ageing days. Anthocyanins can improve VOCs and sensory profile of lamb meat. [ABSTRACT FROM AUTHOR]

Simple Summary: The role of miR-148a-3p in colorectal cancer (CRC) is still debated. In this study, the in vitro antineoplastic effect of miR-148a-3p overexpression in the CRC model is reported. The antitumor activity of miR-148a-3p occurs through apoptosis, mitochondrial impairment, lipid peroxidation, and ferroptosis sustained by the ACSL4/TFRC/Ferritin axis. Bioinformatic analysis and transfection experiments with miR-148a-3p mimics and inhibitors revealed that the cytotoxicity might be related to the downregulation of SLC7A11. These findings, extending knowledge on functional and molecular mechanisms, unveil the oncosuppressor role of miR-148a-3p, pointing out its potential as a diagnostic and therapeutic biomarker in CRC. Ferroptosis, an iron-dependent form of cell death, and dysregulated microRNA (miRNA) expression correlate with colorectal cancer (CRC) development and progression. The tumor suppressor ability of miR-148a-3p has been reported for several cancers. Nevertheless, the role of miR-148a-3p in CRC remains largely undetermined. Here, we aim at investigating the molecular mechanisms and regulatory targets of miR-148a-3p in the CRC cell death mechanism(s). To this end, miR-148a-3p expression was evaluated in SW480 and SW620 cells and normal colon epithelial CCD 841 CoN cells with quantitative real-time polymerase chain reaction (qRT-PCR). Data reported a reduction of miR-148a-3p expression in SW480 and SW620 cells compared to non-tumor cells (p < 0.05). Overexpression of miR-148a selectively inhibited CRC cell viability (p < 0.001), while weakly affecting normal CCD 841 CoN cell survival (p < 0.05). At the cellular level, miR-148a-3p mimics promoted apoptotic cell death via caspase-3 activation (p < 0.001), accumulation of mitochondrial reactive oxygen species (ROS) (p < 0.001), and membrane depolarization (p < 0.001). Moreover, miR-148a-3p overexpression induced lipid peroxidation (p < 0.01), GPX4 downregulation (p < 0.01), and ferroptosis (p < 0.01), as revealed by intracellular and mitochondrial iron accumulation and ACSL4/TFRC/Ferritin modulation. In addition, levels of SLC7A11 mRNA and protein, the cellular targets of miR-148a-3p predicted by bioinformatic tools, were suppressed by miR-148a-3p's overexpression. On the contrary, the downregulation of miR-148a-3p boosted SLC7A11 gene expression and suppressed ferroptosis. Together, these in vitro findings reveal that miR-148a-3p can function as a tumor suppressor in CRC by targeting SLC7A11 and activating ferroptosis, opening new perspectives for the rationale of therapeutic strategies through targeting the miR-148a-3p/SLC7A11 pathway. [ABSTRACT FROM AUTHOR]

Canine mammary tumours (CMTs) are the most common cancer in intact female dogs. In addition to surgery, additional targeted and non-targeted therapies may offer survival benefits to these patients. Therefore, exploring new treatments for CMT is a promising area in veterinary oncology. CMT cells have an altered lipid metabolism and use the oxidation of fatty acids for their energy needs. Here we investigated the tumoricidal effects of teglicar, a reversible inhibitor of carnitine palmitoyl transferase 1A (CPT1A), the rate-limiting enzyme for fatty acid import into mitochondria, on two CMT cells, P114 and CMT-U229. Viability and apoptosis were examined in CMT cells using the crystal violet assay, trypan blue assay, and flow cytometry analysis. The expression of mediators of apoptosis signalling (e.g., caspase-9, caspase-8, and caspase-3) was assessed by quantitative real-time polymerase chain reaction and western blot analyses. Teglicar was able to decrease cell viability and induce apoptosis in P114 and CMT-U229 cells. At the molecular level, the effect of teglicar was associated with an upregulation of the mRNA expression levels of caspase-9, caspase-8, and caspase-3 and an increase in their protein levels. In summary, our results show that teglicar has a potential effect against CMTs through the induction of apoptotic cell death, making it a promising therapeutic agent against CMTs. [ABSTRACT FROM AUTHOR]

In the present study, we aimed at assessing the influence of breed and feeding system on the bovine milk profile of betaines and carnitines and milk capacity in counteracting the inflammatory endothelial cell (EC) damage induced by interleukin (IL)-6. In the first experimental design, two breeds were chosen (Holstein vs. Modicana) to investigate the biomolecule content and antioxidant capacity in milk and dairy products. In the second experimental design, two feeding systems (pasture vs. total mixed ratio) were tested only in Holstein to evaluate the possible effect on the functional profile of milk and dairy products. Finally, the bulk milk from the two experimental designs was used to evaluate the efficacy of preventing IL-6-induced endothelial inflammatory damage. Results showed that Modicana milk and whey had higher biomolecule content and antioxidant activity compared to Holstein milk (p < 0.01). Milk from Holstein fed TMR showed higher concentration of γ-butyrobetaine, δ-valerobetaine (p < 0.01), and l-carnitine (p < 0.05). Similarly, whey from Holstein fed TMR also showed higher content of δ-valerobetaine, glycine betaine, l-carnitine, and acetyl-l-carnitine (p < 0.01) compared to the Holstein fed pasture. Conversely, the antioxidant activity of milk and dairy products was not affected by the feeding system. In ECs, all milk samples reduced the IL-6-induced cytokine release, as well as the accumulation of reactive oxygen species (ROS) and the induction of cell death, with the most robust effect elicited by Modicana milk (p < 0.01). Overall, Modicana milk showed a higher content of biomolecules and antioxidant activity compared to Holstein, suggesting that the breed, more than the feeding system, can positively affect the health-promoting profile of dairy cattle milk. [ABSTRACT FROM AUTHOR]

The aim of this study was to quantify nitrogen (N) and phosphorus (P) utilisation and excretion in intensive dairy buffaloes. Italian Mediterranean buffaloes were divided into 4 groups (n=6/group) as follows: Groups M50, M125 and M225 at 50, 125 and 225 days of lactation, respectively, and Group NL which was non-lactating. Lactating buffaloes had greater (P<0.05) dry matter and organic matter intake than NL buffaloes. Buffaloes in Group M50 had a lower (P<0.05) crude protein (CP) apparent digestibility than buffaloes in Groups M125 and M225. Group NL had a higher (P<0.01) real CP digestibility than Group M50. Buffaloes in Groups M50 and NL had a negative N balance (-79 and -12 g, respectively) whilst buffaloes in Groups M125 and M225 had a positive N balance (26 and 16 g, respectively). Buffaloes in Group M50 had more (P<0.05) N in urine (204 g) than Group NL (87 g). Buffaloes in Group NL had less (P<0.01) P in both faeces (12.9 g) and urine (0.8 g) compared with the three groups of lactating buffaloes combined (25 g in faeces and 12 g in urine) and they also had greater (P<0.05) P digestibility (68%) than lactating buffaloes combined (43%). The present study has shown that buffaloes have a relatively high efficiency of N and P utilisation. [ABSTRACT FROM AUTHOR]