Natural antioxidants are widely investigated as potential substitutes for synthetic compounds suitable for use in organic diets. They are often included in diets based on literature and manufacturer recommendations, sometimes leading to inconsistent results. In this study, we replaced 0.05 g/kg of vitamin E in poultry diets (vitE) with commercial products containing grape skin and green tea extracts (GSGT) or hydrolyzed wood polyphenols (HWP), at doses calculated based on their in vitro antioxidant capacity measured by the 2.2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) test (ABTS). A total of 252 one-day-old male Ross 308 chicks, reared according to organic farming guidelines, were assigned to 9 pens in a randomized block design with 3 replicates per dietary treatment; 108 were slaughtered at 42 days as in the conventional production cycle, while the remaining were slaughtered at 84 days, in accordance with organic farming regulations. Overall, no differences were observed in performance, antioxidant capacity, hematological parameters, carcass and cut weights, meat quality, and survival rate among the dietary treatments. The control group had the highest plasma vitamin E levels (p < 0.0001), while at 42 days, the HWP diet increased plasma vitamin A (p = 0.041) and reduced ALT-GPT levels (p = 0.008). The GSGT diet reduced creatinine levels (p < 0.0001) and showed higher meat protein content compared to HWP (p = 0.024). Differences in gut histomorphology were observed at 42 and 84 days, with effects on specific intestinal regions. Natural antioxidants modulated immune-related gene expression, maintaining the innate immune system in an “alert” state compared to control. The GSGT and vitE groups showed the highest European Production Efficiency Factor (EPEF) at 42 days, while at 84 days, a higher EPEF was observed for the HWP and GSGT groups. In conclusion, HWP and GSGT can effectively replace vitamin E in poultry diets when balanced for antioxidant capacity, in both conventional and organic diets.

Nitrogen content in urine plays a crucial role in assessing the environmental impact of dairy farming. Urine acidifications avoid urine nitrogen volatilization, but potentially lead to a degradation of creatinine, the most dependable marker for quantifying total urine excretion volume, affecting its measurement. This study aimed to assess how acidifying urine samples affects the concentration and detection of creatinine in dairy cattle. In this trial, individual urine samples from 20 Holstein lactating dairy cows were divided into three subsamples, allocated to 1 of 3 groups consisting of 20 samples each. Samples were immediately treated as follows: acidification with H₂SO₄ (1 mL of acid in 30 mL of sample) to achieve a pH < 2 (Group 1)); addition of an equal volume of distilled water (1 mL of distilled water in 30 mL of sample) to investigate dilution effects (Group 2); or storage without any acid or water treatment (Group 3). An analysis of creatinine levels was carried out using the Jaffe method. The Friedman test was employed to compare urine groups across treatments, and the Bland–Altman test was used to assess the agreement between measurements in Group 1 and Group 3. Urinary creatinine values were statistically different (p < 0.001) between Group 1 (median 48.5 mg/dL; range 36.9–83 mg/dL), Group 2 (median 47.5 mg/dL; range 36.5–80.7 mg/dL), and Group 3 (median 48.9 mg/dL, range 37.2–84). Bland–Altman analysis demonstrates agreement between Group 3 and Group 1. The measurement of urinary creatinine using the Jaffe method is affected by sample acidification, but the use of creatinine as a marker for total urine output could remain a viable tool when urine samples are acidified.

A short-term study was conducted to compare the effect of using poplar wood chips (PWC) instead of wheat straw (WS) litter in dairy cows. A total of 38 lactating Holstein cows (204 ± 119 days in milk, 26.9 ± 6.5 kg of milk yield [MY]) were housed in a tiestall farm for a 10-d trial including 5 d of adaptation followed by 5 sampling days (from d 5 to 10). Cows were divided into 2 homogeneous groups: one group was bedded with WS, and the second with PWC. Both litter materials were provided in the amount of 7 kg/stall per d. Each group was composed of 3 subgroups of 6 or 7 cows; the subgroups were physically separated along the feeding line by wooden boards. During the sampling days, fecal composition, used litter composition, and bacterial count (Clostridium spp., Salmonella spp., Escherichia coli, Lactobacillus, and total bacterial count) were analyzed by subgroup twice a day. On d 1 and from d 5 to 10, udder hygiene score and cow cleanliness score were also evaluated individually twice a day. Meanwhile MY, milk hygiene (total bacterial count [TBC], coliform bacterial count [CBC], and spore-forming unit [SFU]) and quality were measured and analyzed from 9 animals per group. Moreover, individual animal behavior (body position and behavioral traits) and subgroup dry matter intake were measured on d 9 and 10. Fecal dry matter did not differ between groups, PWC had the lowest used litter moisture and N content favoring the highest clean cow frequency, but also gave rise to the greatest used litter microbial contamination. The MY, milk quality, TBC, SFU, and CBC were similar. The lying behavior frequency was similar between groups. However, the PWC group showed the lowest sleeping frequency, the highest frequency of other behaviors (including discomfort signs), and the lowest dry matter intake. However, despite this apparent reduction in cow comfort, no biologically important differences were observed in this short-term study between cows on PWC and WS in milk production or hygiene.

Nerolidol (NER) is a sesquiterpene alcohol with recognized antimicrobial potential, whose applications as pure substance are limited by hydrophobicity, instability, and cytotoxicity. Invasomes, i.e. liposomes with terpene ingredients, offer a strategy to improve its delivery; however, the NER loading limits compatible with vesicle integrity are still unclear. Here, Nerolidol-loaded invasomes were produced using a controlled simil-microfluidic coaxial injection process. As a preliminary step, unloaded liposomes were fabricated to consolidate operating conditions and ensure their reproducible colloidal properties. Thereafter, formulations with progressively decreasing nominal NER loads were investigated to evaluate vesicle size, polydispersity, ζ-potential, encapsulation efficiency, effective loading, and stability. High nominal loads promoted turbidity, size increase (by agglomeration coalescence phenomena), and structural instability, whereas formulations containing approximately 1–2% NER achieved nearly complete encapsulation, Z-average ≈ 300 nm, |ζ| > 30 mV, and satisfactory physical stability. Antimicrobial and cytotoxic profiles of representative formulations, previously evaluated in an independent study, confirmed biological activity. Overall, this work identifies a realistic loading window for Nerolidol invasomes and highlights the suitability of the simil-microfluidic approach to obtain scalable, well-controlled formulations, providing a rational basis for their future biological assessment. Nerolidol invasomes systems indeed can be considered a promising versatile platform for antimicrobial applications, including prospective use in animal feed.