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Abstract Antimicrobials are extensively used in livestock to treat common diseases, including mastitis and other bacterial infections, with around 80% of food-producing animals receiving such treatments. Despite a ban in the EU since 2006, antibiotics are still used globally for growth promotion, exacerbating antimicrobial resistance (AMR). From the consumers’ point of view, the potential exposure to resistant zoonotic bacteria in animal-derived food products may represent a threat to public health. However, consumers’ knowledge and attitude vary across socio-demographic characteristics and geographical location. This study aims to evaluate the knowledge, attitudes, and perceptions (KAP) of milk consumers regarding AMR in Pakistan (as a representative of low-to-middle income countries (LMICs)), compared to Italy (as a representative of high-income countries (HICs)). Data were collected via surveys from 401 Pakistani and 550 Italian milk consumers, focusing on socio-demographics, milk purchasing habits, and awareness of antibiotic residues and AMR. The study identified three principal components (PCs) influencing milk consumption: milk quality attributes and tradition, family-dependence, and eating style-dependence. A cluster analysis categorized consumers into four groups: "Attentive to milk quality attributes," "Loyal to milk," "Undecided consumer," and "Milk is essential in my food pattern." Italian consumers predominantly belonged to the "Attentive to milk quality attributes" group, valuing safety and health benefits, and were the most informed about AMR. Pakistani consumers, primarily belonging to the "Loyal to milk" group, exhibited high purchasing frequency influenced by family size but showed limited awareness about AMR risks. The "Undecided consumer" group, a mix of Italian and Pakistani individuals, lacked strong preferences and displayed moderate awareness of AMR, influenced by habitual consumption patterns. This research underscores the need for targeted educational campaigns to raise awareness about AMR and antibiotic residues in milk, particularly in LMICs. Survey findings can help to identify targeted action plans and boost policy development on AMR. Enhancing consumer knowledge through the One-Health approach could mitigate AMR risks and promote informed decision-making in milk consumption across different socio-economic contexts.

The most frequent adverse effects of AFB1 in chicken are low performance, the depression of the immune system, and a reduced quality of both eggs and meat, leading to economic losses. Since oxidative stress plays a major role in AFB1 toxicity, natural products are increasingly being used as an alternative to mineral binders to tackle AFB1 toxicosis in farm animals. In this study, an in vivo trial was performed by exposing broilers for 10 days to AFB1 at dietary concentrations approaching the maximum limits set by the EU (0.02 mg/kg feed) in the presence or absence of turmeric powder (TP) (included in the feed at 400 mg/kg). The aims were to evaluate (i) the effects of AFB1 on lipid peroxidation, antioxidant parameters, histology, and the expression of drug transporters and biotransformation enzymes in the liver; (ii) the hepatic accumulation of AFB1 and its main metabolites (assessed using an in-house-validated HPLC-FLD method); (iii) the possible modulation of the above parameters elicited by TP. Broilers exposed to AFB1 alone displayed a significant increase in lipid peroxidation in the liver, which was completely reverted by the concomitant administration of TP. Although no changes in glutathione levels and antioxidant enzyme activities were detected in any treatment group, AFB1 significantly upregulated and downregulated the mRNA expression of CYP2A6 and Nrf2, respectively. TP counteracted such negative effects and increased the hepatic gene expression of selected antioxidant enzymes (i.e., CAT and SOD2) and drug transporters (i.e., ABCG2), which were further enhanced in combination with AFB1. Moreover, both AFB1 and TP increased the mRNA levels of ABCC2 and ABCG2 in the duodenum. The latter changes might be implicated in the decrease in hepatic AFB1 to undetectable levels (

An ad hoc questionnaire was designed in order to investigate AMR knowledge amongst Italian dog owners, owner expectations concerning pharmacological treatment of canine AD, and client attitudes towards and compliance with alternative strategies to antimicrobial administration. A total of 250 questionnaires were returned. Most of respondents were female, aged 36–70 and workers. More than a half of participants owned one dog with mixed breed, with Labrador retriever, golden retriever, dachshund, and border collie being the most represented breeds. On average, each dog was treated with an oral antibiotic 1.044 times per year. Intestinal diseases were among the main reasons (19%) for antibiotic prescription. Oral antibiotic courses without veterinary consultation (21%) and anticipated termination of the therapy (17.1%) were less common than reported elsewhere. The majority of respondents knew the meaning of AMR with a significant inverse association between the level of education and the tendency to administer antimicrobials without consulting a clinician (p = 0.004). Most of the owners expected a rapid recovery of clinical signs after a first episode of AD and accepted natural dietary supplementation for treating the condition. Ninety-five percent of the respondents believed that public funding should be spent to study AMR. Even though an acceptable degree of AMR awareness emerged, we feel that further efforts should be made to increase public AMR knowledge and to stimulate proactive measures to fight the phenomenon. On the other hand, the development of guidelines for the treatment of uncomplicated canine AD would help clinicians to rationalize antimicrobial use.

We report a case of laboratory-confirmed Campylobacter (C). jejuni and C. upsaliensis symptomatic infection in a puppy, a French Bouledogue, female, 6 months of age, fed a raw, unbalanced, poultry-based diet (RPD), (48.1 CP, 33% EE, 0.3% Ca, 0.5% Phos, 0.5 Ca/P, on a dry-matter basis), and in its owner. Soon after adoption, the pet and the caregiver showed severe gastrointestinal signs and needed hospitalization. Fecal PCR assays, selective cultures, and antibiotic susceptibility testing were performed, and multi-drug resistant C. jejuni and C. upsaliensis were isolated from the feces of both. The same bacteria were also identified by FISH in the dog colonic biopsies collected during endoscopy. The puppy was prescribed a complete commercial diet for growing dogs, (30.00% CP, 21.00% EE, 1.2% Ca; 1% Phos; as fed) and treated with ciprofloxacin. The dog and the man healed uneventfully and tested negative for further fecal PCR analyses. This report focuses on dog nutritional management and explores the potential routes of exposure, with emphasis on emerging outbreaks related to current pet food fads. Our data support the One Health approach, where veterinarians, physicians, and owners are challenged to build effective stewardship to prevent the spread of zoonoses.

Aflatoxin B1 (AFB1) causes hepatotoxicity, immunotoxicity, and kidney damage, and it is included in group I of human carcinogens. The European Commission has established maximum limits of AFB1 in feed, ranging from 5 to 20 μg/kg. Chicken is moderately sensitive to AFB1, which results in reduced growth performance and economic losses. Oxidative stress triggered by AFB1 plays a crucial role in kidney damage and the antioxidant activity of Curcumin (CURC) could help in preventing such adverse effect. Twenty-days-old broilers were treated for 10 days with AFB1 (0.02 mg/kg feed), alone or in combination with CURC (400 mg/kg feed), to explore the effects on the renal tissue. Animals exposed to AFB1 alone displayed alterations of the oxidative stress parameters compared with controls: serum antioxidant capacity, and enzymatic activity of kidney superoxide dismutase, catalase and glutathione peroxidase were decreased, while renal malondialdehyde levels and NADPH oxidase complex expression were increased. The administration of CURC attenuates all the oxidative stress parameters modified by AFB1 in the chicken kidney, opening new perspectives in the management of aflatoxicosis.

Abstract Background Feline hyperthyroidism, the most common endocrinopathy in older cats, provides a spontaneous model for human thyrotoxicosis. Human thyrotoxicosis is associated with redox unbalance, which may result in organ damage. The redox status of hyperthyroid cats is largely unknown. The aims of the present study were to compare the redox status of cats with hyperthyroidism with that of healthy cats and cats with chronic non-thyroidal illness. Results Forty cats with untreated hyperthyroidism (group H), 45 chronically ill cats with non-thyroidal illness (group I), and 39 healthy cats (group C) were recruited for this observational cross-sectional study. All cats were screened for redox status markers. Determinable reactive oxygen metabolites (d-ROMs) were used as oxidative stress markers. Antioxidant status was determined using the OXY-Adsorbent test to quantify the plasma barrier to oxidation. The Oxidative Stress index (OSi) was calculated as the ratio of d-ROMs and OXY-Adsorbent test values. Data were compared by ANOVA with Tukey’s multiple comparisons post-hoc test. The dROMs of group H (193 ± 47 CarrU) were significantly higher (p

Canine acute enteropathies (AE) are common morbidities primarily managed with supportive therapy. However, in some cases, unnecessary courses of antibiotics are empirically prescribed. Recent studies in humans have hypothesized the use of antioxidants as a possible alternative and/or support to antimicrobial drugs in uncomplicated cases. Considering the global need to reduce the antibiotic use, the aim of the study was to compare the oxidative burden of the diarrhetic population to that of healthy dogs. Forty-five patients suffering from uncomplicated acute diarrhea (AD) and 30 controls were screened for clinical and biochemical parameters, and serum redox indices (reactive oxygen metabolites, dROMs; serum antioxidant capacity, SAC; oxidative stress index, OSi). The average levels of dROMs in AD dogs were significantly higher (p < 0.001) than in healthy dogs, while SAC did not significantly differ between the two groups. However, the OSi values (ratio between dROMs and SAC) significantly increased (p < 0.001) in AD dogs compared to controls. The study demonstrates that canine AD could induce redox imbalance. Although its role in the etiopathogenesis and evolution of the disease should be further investigated, our results suggest that the improvement of the patient oxidative status, possibly through the dietary administration of antioxidants, could support the management of canine AE, reducing the use of antibiotics.

Due to the climatic change, an increase in aflatoxin B1 (AFB1) maize contamination has been reported in Europe. As an alternative to mineral binders, natural phytogenic compounds are increasingly used to counteract the negative effects of AFB1 in farm animals. In cows, even low dietary AFB1 concentrations may result in the milk excretion of the genotoxic carcinogen metabolite aflatoxin M1 (AFM1). In this study, we tested the ability of dietary turmeric powder (TP), an extract from Curcuma longa (CL) rich in curcumin and curcuminoids, in reducing AFM1 mammary excretion in Holstein–Friesian cows. Both active principles are reported to inhibit AFM1 hepatic synthesis and interact with drug transporters involved in AFB1 absorption and excretion. A crossover design was applied to two groups of cows (n = 4 each) with a 4-day washout. Animals received a diet contaminated with low AFB1 levels (5 ± 1 µg/kg) for 10 days ± TP supplementation (20 g/head/day). TP treatment had no impact on milk yield, milk composition or somatic cell count. Despite a tendency toward a lower average AFM1 milk content in the last four days of the treatment (below EU limits), no statistically significant differences with the AFB1 group occurred. Since the bioavailability of TP active principles may be a major issue, further investigations with different CL preparations are warranted.

The treatment of choice for feline hyperthyroidism is the administration of the antithyroid drug methimazole. Both the endocrinopathy and the drug adverse reactions (e.g., hepatotoxicosis, gastrointestinal disorders, and renal injury) are partly due to oxidative stress and redox unbalance. This study investigated the free radical production and the impairment of the antioxidant barrier induced by methimazole in an in vitro model of feline renal epithelium. The protective effects of quercetin and resveratrol were also explored. CRFK cells were incubated with a methimazole concentration equivalent to the maximum plasma levels in orally treated cats (4 µM), in the presence or absence of either one of the two selected antioxidants at different time-points (up to 72 h). Cell viability, ROS production, GSH levels, and mRNA expression of antioxidant enzymes (i.e., CAT, SOD, GPx, and GST) were assessed. Methimazole impaired cell viability and increased ROS levels in a time-dependent manner. Similarly, GSH content and CAT, SOD, and GPx3 expression were higher compared with control cells. Such effects were significantly counteracted by quercetin. These results provide new insights about the mechanisms underlying the methimazole-related side effects frequently observed in hyperthyroid cats. They also support the use of quercetin in the management of feline hyperthyroidism.

Feline hyperthyroidism is a clinical syndrome related to an excessive production of thyroid hormones, and it is considered as a spontaneous animal model for human thyrotoxicosis. Many shared features between the feline and the human disease have been identified so far, including pathogenesis, clinical signs, and treatment options. Although methimazole is considered the first-choice drug in both species, several side effects have been described. Furthermore, methimazole could interfere with the oxidative status, already perturbated by the disease. It has been reported in humans that dietary management, mainly through antioxidant supplementation, could mitigate this oxidative burden. The purpose of the review is to describe current therapeutic options in the course of feline hyperthyroidism and to summarize the state of the art relationship between dietary antioxidants administration and the reduction of methimazole side-effects in humans to support the use of this approach also in cats.

Ochratoxin A (OTA) is a powerful mycotoxin found in various foods and feedstuff, responsible for subchronic and chronic toxicity, such as nephrotoxicity, hepatotoxicity, teratogenicity, and immunotoxicity to both humans and several animal species. The severity of the liver damage caused depends on both dose and duration of exposure. Several studies have suggested that oxidative stress might contribute to increasing the hepatotoxicity of OTA, and several antioxidants, including curcumin (CURC), have been tested to counteract the toxic hepatic action of OTA in various classes of animals. Therefore, the present study was designed to evaluate the protective effect of CURC, a bioactive compound with different therapeutic properties on hepatic injuries caused by OTA in rat animal models. CURC effects were examined in Sprague Dawley rats treated with CURC (100 mg/kg), alone or in combination with OTA (0.5 mg/kg), by gavage daily for 14 days. At the end of the experiment, rats treated with OTA showed alterations in biochemical parameters and oxidative stress in the liver. CURC dosing significantly attenuated oxidative stress and lipid peroxidation versus the OTA group. Furthermore, liver histological tests showed that CURC reduced the multifocal lymphoplasmacellular hepatitis, the periportal fibrosis, and the necrosis observed in the OTA group. This study provides evidence that CURC can preserve OTA-induced oxidative damage in the liver of rats.

Acute diarrhea is one of the most common reasons why pet owners seek veterinary care for their canine companions. In many cases, signs resolve spontaneously or with symptomatic therapy without a specific cause being discovered. However, life-threatening cases can occur. The etiology is complex, including infectious diseases (endoparasites, virus, bacteria, protozoa, fungal agents) by both zoonotic and non-zoonotic pathogens, dietary indiscretion, endocrine diseases, and stress (e.g., travel or environmental changes). In the last years, the role played by oxidative stress in the pathogenesis of acute and chronic enteropathies, independently from the initial noxa, has been highlighted by many researches in both humans and animals. As a result, a series of dietary antioxidant compounds have been studied for their potential use in the treatment of intestinal inflammation. This review summarizes the traditional therapeutic and nutritional options to manage canine acute diarrhea, highlighting the need to explore the role of oxidative stress and potential antioxidant supplementation, especially polyphenols, during acute diarrheic episodes.

Canine aggression is a major concern, affecting millions of people worldwide, and treatment can be challenging even for skilled veterinarians. Empiric use of fluoxetine is sometimes attempted, although few data regarding long-term effects in aggressive dogs are available. The aim of the study was to investigate clinical effectiveness of fluoxetine (1.5 mg/kg/die PO) combined with a behavior modification program for treatment of canine dominance-related aggression. Circulating levels of fluoxetine, norfluoxetine, and serotonin (5-HT) were also measured. Eight dogs with a diagnosis of dominance aggression (owner-directed) were enrolled. Before treatment (T0), and after one (T1), two (T2), four (T3), and six (T4) months of fluoxetine administration, clinical outcomes were graded using a five-point frequency scale (0–4), and blood samples were collected to measure fluoxetine/norfluoxetine (high-performance liquid chromatography) and 5-HT (ELISA) levels. Following treatment, a decrease in behavioral test scores was observed at T1–T4. Increasing concentrations of circulating fluoxetine and norfluoxetine were measured throughout the follow-up. Correlation between norfluoxetine levels and clinical scores was observed at T4. Starting from T1, a significant decrease in 5-HT levels was observed. Our data suggest that fluoxetine (1.5 mg/kg/day) when associated with behavior treatment is effective in controlling canine aggression over a six-month period, and that, in dogs norfluoxetine levels seem reliable in predicting clinical efficacy.

Methimazole (MMI) is often the selected medical treatment for feline hyperthyroidism. However, the onset of MMI-related side effects (MMI-SE) is likely caused by oxidative stress. This study evaluated the dietary supplementation of selected antioxidants in hyperthyroid cats receiving MMI, to reduce MMI-SE. Thirty hyperthyroid client-owned cats were randomly allocated in group M (MMI + placebo) or group M+A (MMI + antioxidants). At different time-points from the enrolment (ET) to the end of the trial (FT), the following information was recorded: clinical findings, complete blood count, serum biochemical parameters, urinalysis, total plasma thyroxine concentrations, determinable reactive oxygen metabolites (dROMs), OXY-adsorbent test values, and oxidative stress index (OSi) values, and MMI-SE. dROMs and OSi values significantly increased from ET to FT in group M and were significantly higher in group M than in group M+A at FT. Likewise, OXY-absorbent test values were significantly higher in group M+A than in group M at FT. Moreover, the occurrence rate of MMI-SE in group M+A was lower than in group M. In conclusion, our results show that the dietary supplementation of antioxidants in hyperthyroid cats receiving MMI exerts a protective effect against oxidative stress, likely contributing to the reduction of MMI-SE.

Our archaeological and anthropological investigations carried out inside the Crypt of the Franciscan Monastery in Azzio (Varese, Northern Italy) allowed us to discover a singular funerary practice of Franciscan friars. It consisted of a secondary burial practice.

The cynomolgus monkey (Macaca fascicularis) is a well-known non-human primate species commonly used in non-clinical research. It is important to know basal clinical pathology parameters in order to have a reference for evaluating any potential treatment-induced effects, maintaining health status among animals and, if needed, evaluating correct substantiative therapies. In this study, data from 238 untreated cynomolgus monkeys (119 males and 119 females of juvenile age, 2.5 to 3.5 years) kept under laboratory conditions were used to build up a reference database of clinical pathology parameters. Twenty-two hematology markers, 24 clinical chemistry markers and two blood coagulation parameters were analyzed. Gender-related differences were evaluated using statistical analyses. To assess the possible effects of stress induced by housing or handling involved in treatment procedures, 78 animals (35 males and 35 females out of 238 juvenile monkeys and four adult males and four adult females) were used to evaluate cortisol, corticosterone and behavioral assessment over time. Data were analyzed using a non-parametric statistical test and machine learning approaches. Reference clinical pathology data obtained from untreated animals may be extremely useful for investigators employing cynomolgus monkeys as a test system for non-clinical safety studies.