In recent years, the increasing cost of feed ingredients and the need for environmental sustainability in the poultry sector have necessitated the exploration of alternative feed sources. In this context, the production of hydroponic green fodder (HGF) is attracting attention, as it is an environmentally friendly method that minimizes water and land use. It has been reported that hydroponic fodder can positively impact production performance, welfare, and digestive health, particularly in poultry. This study was conducted to investigate the effects of HGF supplementation in the last quarter of the growing season on growth performance, carcass, and meat quality traits in the slow-growing Gerze Turkish native chicken breed. In the study, chicks were fed starter feed until 8 weeks of age, grower feed between 8 and 14 weeks, and either finisher feed alone (control group, CD) or finisher feed plus HGF (treatment group) ad libitum between 14 and 18 weeks of age. At the end of the trial, the CD group was found to have significantly higher abdominal fat weight, dry matter percentage, and protein levels than the HGF group (p < 0.05). In contrast, ash content was lower in both the pectoralis major (PM) and gastrocnemius (GN) muscles in the control group (p<0.05). In chickens who were fed HGF, drip loss and intramuscular fat content in the GN muscle on the second storage day increased significantly (p<0.05), while cooking loss in the GN muscle and freezing loss in the PM muscle decreased (p<0.05). Furthermore, the hardness and adhesiveness values ​​of the PM muscle in the CD group were higher than in the HGF group (p<0.05).
In conclusion, it was determined that the use of hydroponic green feed can improve meat quality traits in Gerze chickens without negatively affecting growth performance. These findings suggest that HGF is a potential alternative feed source in sustainable and efficient poultry farming practices.

Nutrition plays a vital role in regulating reproductive performance in humans and animals. Recently, the inclusion of agro-industrial by-products such as noodle waste (NW) in animal diets has increased, often without the adequate evaluation of their potential reproductive consequences. This study investigated the effects of NW consumption on ovarian function in twenty (20) healthy adult female Wistar rats. The rats were randomly assigned to four groups (n = 5). Group A (control) received standard rat feed, while Groups B, C, and D were fed diets containing NW only, 50% NW supplemented with 50% fish meal (FM), and 30% NW supplemented with 70% FM, respectively, for 35 days. Serum samples were analyzed for reproductive hormones (progesterone, oestradiol, and follicle-stimulating hormone (FSH)) using an enzyme-linked immunosorbent assay (ELISA). Ovaries were excised to evaluate ovarian biometry, gonadosomatic index (GSI), oxidative stress markers, antioxidant enzymes, and histopathology.

There were no significant differences (p > 0.05) in the mean serum levels of FSH, progesterone, and oestradiol among the treatment groups. Similarly, mean malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) levels showed no significant variation (p > 0.05), although Group B (NW only) exhibited higher MDA and antioxidant enzyme levels. Significant differences (p < 0.05) were observed in right ovarian weight (Group B: 0.03 ± 0.01 g and Group C: 0.06 ± 0.02 g), left ovarian weight (Group B: 0.02 ± 0.01 g and Group C: 0.06 ± 0.01 g), and GSI (Group B: 0.03 ± 0.02 and Group C: 0.07 ± 0.00). Histological examination revealed extensive follicular atresia, reduced ovarian vascularization, and smaller ovarian size in Group B compared to other groups.

These findings suggest that exclusive consumption of noodle waste may impair ovarian function through oxidative stress and structural degeneration, emphasizing the need for appropriate dietary supplementation to mitigate its adverse effects.

Pesticide exposure is a critical concern for honeybee health. Flupyradifurone (FPF) is widely used in agriculture but has been associated with neurotoxic effects in honeybees (Apis mellifera), including impaired motor coordination, hyperactivity, and lethargy. Given the growing need for protective strategies, natural antioxidants such as polyphenolic extracts have gained attention for their potential to mitigate pesticide-induced toxicity. Pistacia lentiscus has demonstrated anti-inflammatory and antioxidant properties in mammalian models, but its protective role in honeybees remains underexplored. This study aimed to evaluate whether Pistacia lentiscus leaf polyphenolic extract (LPE) could reduce FPF-induced toxicity in honeybees. Honeybees were allocated into 12 experimental groups (n=20/group) under controlled laboratory conditions. FPF was administered orally at 100 mg/L for 72 hours, while parallel groups received co-administration of LPE (1% or 5% w/v). Clinical observations were recorded daily, including neuromuscular dysfunction (e.g., tremors, abnormal wing positioning, uncoordinated movement, curved belly, etc.), gastrointestinal distress (e.g., distended abdomen, abnormal and/or defecation), behavioral alterations (e.g., hyperactivity followed by apathy, failure to respond to stimuli), and mortality rates. Honeybees exposed to FPF alone exhibited severe neurotoxic symptoms within 24 hours, including erratic movement (67.5% incidence) and moribund posturing (27.5% by Day 3), with 90% mortality by 72 hours. In contrast, co-treatment with LPE at 1% significantly delayed symptom progression and prevents mortality: FPF +1% LPE groups showed 100% survival at 72 hours vs 10% in FPF-only groups (p<0.01). The FPF +1% LPE groups showed lesser hyperactivity (2,5% vs. 67.5% in FPF-only) and abdominal spasms (0% vs. 27.5%). FPF +5% LPE groups also showed higher survival probability but in a lesser amount (42.5% vs. 10% FPF-only), suggesting that high LPE doses are toxic. The therapeutic activity is attributable to the polyphenolic component identified through phytochemical characterisation using HPLC-MS. LPE attenuated FPF-induced toxicity and should be considered for further field studies to confirm its therapeutic potential.

Bisphenols (BPs) are chemical compounds largely used in the plastic industry and in several manufacturing processes. Their presence has been documented in several environmental and biological matrices. Many of them have been found to act as endocrine disruptors, causing negative effects on animals and humans. Bisphenol A is the precursor of all bisphenols, and given its documented toxicity, it has been banned and substituted by several analogues in different processes. Among them, Bisphenol S has been used worldwide, and its presence is reported in several matrices. Nevertheless, these alternative bisphenols do not seem to be safer than their congener. Given these premises, it is of great concern to monitor BPS presence in bivalve mollusks, which can be used as sentinels of environmental contamination. BPs are generally quantified using chromatographic techniques interfaced with high-resolution spectrometers. This study aims to identify a simple and reproducible method able to determine the BPS presence in mussels and to quantify it. For this purpose, a competitive ELISA method was validated and applied to the determination of BPS in mussels. The validated method was applied to the determination of BPS in mussels harvested along Sardinian coasts in Italy, and a mean concentration of 10 ng/g ww of BPS was found. The results of the present study are of great concern from a One Health perspective, given that they reflect the healthiness of the environment and of mussels collected in Sardinia, where over 13,000 tons of mussels are produced every year.

In recent years, new global health threats have been linked to climate change and vector-borne diseases; West Nile viruses (WNV) and Usutu virus (USUV) are Flaviviridae viruses, involved in several outbreaks all over the world. Warmer temperatures and altered precipitation patterns can expand the geographical range and seasonal activity of mosquitoes. Italy has experienced numerous incursions of WNV and USUV, becoming the European country with the highest number of cases. This study aims to present our findings during 4 years of wild bird surveillance for WNV and USUV viruses in the three regions of northwest Italy.

In this study, we carried out both passive (wild birds found dead, syndromic surveillance. and neurological cases in equids) and active surveillance (culling wild birds of target species). After RNA extraction, we performed two RT-PCR to detect WNV Lineage 1 and 2 and USUV.

A total of 1026 animals underwent analysis in 2022; 20 animals tested positive for WNVL2, with only one positive for USUV. In 2023, a total of 1307 animals were tested; 24 animals were positive for WNVL2, 5 were positive for USUV, and 5 were co-infected with WNVL2 and USUV. In 2024, a total of 995 animals underwent testing; 25 animals tested positive for WNVL2. Finally, in 2025, a total of 1184 animals were analyzed; we detected a total of 20-WNVL2 positive samples and 1 WNVL1-positive sample.

Our results show that positive samples are increasingly detected in uncommon periods for these viruses. Currently, it remains difficult to assess the impact of this change on the viruses' dynamics and epidemiology through vectors in relation to the last alternating pattern of periods of the complete absence of rain and subsequent periods of heavy flooding. Finally, we must consider the effect on bird migration and on the balance between viruses and vectors. This is a One-Health concern; public health monitoring requires more integrated plans and actions to anticipate zoonotic viral diffusion.

Introduction

The rapid spread of antimicrobial resistance (AMR) in humans, animals, and the environment poses growing challenges to global health. Rapid, cost-effective methods for routine resistome surveillance in dairy herds are increasingly needed to support antimicrobial stewardship and implement risk management strategies. High-resolution resistome profiling could be a possible solution to better understand the antibiotic resistance landscape in herds related to the dairy industry chain. This study aimed to provide a proof-of-concept for a quantitative analysis of the resistome retrieved from milking filters in six dairy herds in Lombardy (Italy).

Materials and Methods

Milking filters (n=6, one from each herd), replaced after each milking session, were collected. DNA was extracted using the PowerWater DNA kit (Qiagen) and quantified with the NanoReady Touch series Micro Volume Spectrophotometer (Aurogene, Italy). DNA libraries were prepared using the Rapid Barcoding Kit (Oxford Nanopore). Sequencing was performed on a MinION MK1C device.

Microbial communities and resistomes were analyzed through a customized bioinformatic pipeline for taxonomic classification (Centrifuge) and antibiotic resistance gene identification (ABRicate); finally, results were visualized using R statistical software.

Results

The two most abundant families of resistant bacteria were Mycobacteriaceae and Staphylococcaceae for herd 1 and Moraxellaceae and Methylococcaceae for herd 5. Two out of six dairy herds showed high levels of resistance, specifically herd 1 and herd 5 with 335 and 465 genomic copies/Gigabase (gc/Gb) when compared to the other four herds, which showed values lower than 20 gc/Gb. The most predominant levels of resistance were against gentamicin (17.6%) and kanamycin (15.1%) for herd 1 and spectinomycin (37.1%) and tetracyclines (18.8%) for herd 5.

Discussion and Conclusions

This study shows a relatively quick and easy method to quantitatively analyse the resistome of milking filters based on a long-read sequencing approach. This approach can determine precisely the levels of antibiotic resistance present in the sample and associate them with the bacterial families harboring them.

Introduction:
Colic remains a major cause of emergency intervention in horses, and feeding practices are a critical component of risk management. Freshly cut, semi-dried grass is often considered a convenient feed source, yet early fermentation processes may predispose horses to gastrointestinal disturbances. This study investigated microbiological, physicochemical, and sensory changes in grass clippings during natural drying to identify factors contributing to colic development.

Methods:
Grass samples were collected near Stable X during the seasonal peak of grass-related colic. The grass samples were collected at three time points (0 h, after 12 h and 24 h of fermentation), and three independent replicates were obtained at each time point, resulting in a total n=9. Microbiological evaluation was performed using Columbia Blood Agar under anaerobic conditions, followed by Gram staining. Feed quality assessment included dry matter (DM), pH, and concentrations of nitrogen, phosphorus, and calcium. Organoleptic characteristics were also documented. Clinical colic cases recorded in the LSMU Large Animal Clinic during the same period were reviewed to support laboratory findings.

Results:
Across all replicates, Clostridium botulinum was not detected. Instead, Actinomyces spp. and non-specific Gram-positive rods proliferated consistently, with the highest bacterial load in the 24-hour samples. DM displayed a uniform pattern: a slight increase at 12 hours, followed by a statistically significant decline at 24 hours (mean decrease 26.5%, p < 0.05), indicating active microbial fermentation. Nitrogen and phosphorus increased, while calcium decreased; pH remained acidic. Sensory evaluation revealed progressive heating, darkening, and development of spoilage odour. Clinical data demonstrated recurrent episodes of gaseous distension and reduced intestinal motility, consistent with laboratory findings.

Conclusions:
The results confirmed that freshly cut grass undergoes rapid, reproducible fermentation within 24 hours, producing conditions that predispose horses to fermentation-related colic. Semi-dried grass clippings represent a consistent feeding risk and should be avoided. Consider it as a hypothesis and expand the studies.

Introduction:
Equine colic is a multifactorial syndrome, and horse owners frequently relate its occurrence to lunar cycles. Despite the popularity of this belief, scientific data are scarce. This study evaluated whether lunar phases are associated with the frequency of colic episodes in horses and whether they influence colic localisation or clinical outcome.

Methods:
A retrospective analysis was performed using clinical records of horses presenting with colic to a university large animal clinic between 2013 and 2020. Each case was assigned to one of four lunar phases based on the date of onset: new moon, last quarter, first quarter, or full moon. In total, 138 colic episodes with complete data were included. Statistical analyses were carried out in SPSS. A chi-square goodness-of-fit test was used to compare the observed distribution of colic cases across lunar phases with an equal expected distribution. Associations between lunar phase and localisation of colic or disease outcome were examined using chi-square tests of independence. Effect size was estimated using Cohen’s w.

Results:
Colic cases were not evenly distributed across the lunar phases: new moon n=50; last quarter n=39; first quarter n=20; full moon n=29 (χ²=14.52, df=3, p<0.01, w=0.32). Thus, colic episodes occurred significantly more often around new moon and the last quarter than during the first quarter. No significant association was found between the lunar phase and anatomical localisation of colic (p>0.05) or clinical outcome (p>0.05).

Conclusions:
Lunar phases showed a moderate but statistically significant association with the timing of colic episodes, with peaks around new moon and the last quarter. However, moon phases did not affect the type of colic or its outcome. Further prospective studies are needed to clarify underlying mechanisms and to control for management and environmental confounders.

In a stable cat group, interactions such as territorial defence or resource competition may be dynamic or variable. Simultaneous feeding can promote the emergence of aggression when defending or claiming food and represents a useful context to identify changes in hierarchy in pets. The aim of this study was to explore the identification of hierarchical positions occupied by individuals within a group of domestic cats by evaluating the frequency of aggressions performed and received during food offering. Four cats (three females and one male), all neutered and aged between 3 and 6 years, were included. To avoid bias due to modifications in feeding habits, the study was conducted in the home, and behavioural observations were performed by a person already familiar with the cats. Observations were carried out for 14 consecutive days at a fixed schedule for 10 minutes, during which the frequencies of agonistic and aggressive behaviours performed and received by each cat were recorded. Individual aggression indexes were estimated as follows: Aggressions Performed / (Aggressions Performed + Aggressions Received). In addition, aggression frequencies were analysed using the chi-square test. The frequency of aggressions performed and received differed between cats (P < 0.0001), with two cats identified as the main aggressors and one receiving the highest number of aggressions. The cats ranked by aggression index were: youngest female (3 years old) > male (6 years old) > older female (5 years old) > oldest female (6 years old), with an index nearly ten times lower. These results should be interpreted with caution due to the small sample size. We conclude that identifying a high frequency of aggressive behaviours enables the identification of the dominant individual within the group and the lower-ranking animal in the hierarchy, which may present a welfare concern due to reduced control over food access.

Introduction: Recording animal behaviour without bias due to the presence of watchers remains challenging; even if technological advances are helpful, direct observations are still worth it. Habituation can partially alleviate this issue, but for many species, there is still a lack of data. The aim: The aim of this study was to habituate big felids in a zoo to the exposition of visual barriers to obtain a reduced or absence of reaction. Methods: Four tigers (Panthera tigris) and five lions (Panthera leo) from “Parque Centenario” zoo in Mérida, México, were used, all in a good healthy state, and the research protocol was approved by the Bioethics Committee (ref# CB-CCBA-I- 2024-001). On non-continuous days, random and gradual exposition to visual barriers (posters) was made. Posters placed on metal supports were mounted in front of the enclosure and replaced nine times by different people. After each change, one minute of observation plus four of rest were effectuated using focal sampling. On the posters, small square holes covered with mesh allowed the person behind them to observe without being seen. The ethogram considered reaction (i.e., turning ears, body orientation) and no reaction (i.e., absence of movements). Frequencies were weighted by number of watchers and animals by species. A Manova test and Spearman correlation between the frequencies and the progression of days were performed. Results: A total of 12 non-continuous days distributed over 39 days were needed to obtain no reaction of animals. A significant effect (P<0.001, Manova test) of days’ progression was found on the frequency of behaviours studied. Significant negative correlations were found between reaction behaviours (P<0.05, Coef. Corr from -0.36 to -0.74) and days in both tigers and lions. Also, a positive correlation (P<0.05, Coef Corr from 0.87 to 0.95) was found with no reaction behaviours. Conclusion: These big felids can be habituated to visual barriers in a relatively short period of time, enabling hidden observation.