Antimicrobial Use in Cats in a University Veterinary Hospital in Central Italy: A Retrospective Study.

Background: Monitoring antimicrobial use is crucial for understanding current prescribing practices. Such information helps in establishing stewardship programs for effectively using antimicrobials and combating resistance to them. Objectives: This study describes how antimicrobials are prescribed at the Veterinary Teaching Hospital of the University of Pisa and compares how the internal medicine and intensive care units differ in their usage of antimicrobials. Methods: The study involved cats that were presented in the units in 2021 and 2022. Antimicrobial prescriptions were obtained via the hospital's management software (OCIROE). Results: In a population of 1164 non-surgical cats with 397 antimicrobials prescribed, the most prescribed ones were amoxicillin-clavulanic acid in the internal medicine unit and ampicillin in the intensive care unit. Twenty-five percent of all antimicrobials were Highest-Priority Critically Important Antimicrobials or Antimicrobial Advice Ad Hoc Expert Group Category B. The oral route was the main route of administration in the internal medicine unit, while parenteral was the most common route used in the intensive care unit. Most antimicrobials were prescribed to treat pathologic conditions affecting the urinary (39%), gastroenteric (21%), respiratory (13%), and integumentary (12%) systems. A diagnosis, accurate dosage, and the use of species-approved medications were recorded in the antimicrobial prescriptions. However, only 11% of these prescriptions were supported by antimicrobial susceptibility tests. Conclusions: These results suggest room for improvement, particularly in increasing the use of antimicrobial susceptibility testing to ensure more targeted antimicrobial therapy. Given the importance of antimicrobial resistance and the One Health approach, the study also highlights the need to consider the broader impact of antimicrobial use in animals, including the potential contribution to resistance in bacteria that affect both animal and human health.

Computed tomographic features of pulmonary and extrapulmonary lesions can be useful in prioritizing the diagnosis of hemangiosarcoma metastases in dogs.

OBJECTIVE: The aim of this study was to evaluate the CT features of pulmonary metastases in dogs with hemangiosarcoma (HSA) at various sites. Additionally, the CT characteristics of extrapulmonary metastases in the same population were assessed. METHODS: Retrospective, observational, and descriptive study conducted from April 2013 to January 2024. Dogs with histologically confirmed HSA and suspected or cytologically/histologically confirmed lung metastases were included. Dogs were excluded if they had a second primary tumor or only 1 unsampled pulmonary nodule. RESULTS: 33 dogs were included, with 26/33 [78.8%] having more than 10 metastatic pulmonary nodules. Most nodules were generalized (24/33 [72.7%]), miliary (29/33 [87.9%]) to subcentimetric (32/33 [97%]) in size, well-defined margins (29/33 [87.9%]), or a perilesional halo sign (24/33 [72.7%]). When more than 10 nodules were present, a generalized distribution was prevalent, while a peripheral location was more common when 2 to 10 nodules were present (P < .0001). In 32/33 (97%) cases, a pulmonary vessel was directly connected to the nodule (feeding vessel). After contrast administration, most lung metastases appeared homogenous (26/33 [78.8%]), although some showed areas of intense enhancement (5/33 [15.1%]) a feature also observed in extrapulmonary metastases with varying frequency (0% to 85.7%). CONCLUSIONS: Pulmonary HSA metastases were characterized by generalized, small (miliary/subcentimetric), well-defined nodules, commonly associated with a halo sign and feeding vessel. Intralesional areas of spotty postcontrast linear or amorphous strong hyperdensity were frequently observed especially in extrapulmonary metastases. CLINICAL RELEVANCE: These features may help radiologists and clinicians orient their diagnosis toward metastatic HSA.

Interplay of UV-filter pollution and temperature rise scenarios on Mytilus galloprovincialis health: Unveiling sperm quality and adult physiology, biochemistry, and histology insights.

Addressing the impacts of emerging contaminants within the context of climate change is crucial for understanding ecosystem health decline. Among these, the organic UV-filters 4-methylbenzylidenecamphor (4-MBC) and benzophenone-3 (BP-3) are widely used in cosmetics and personal care products. Their unique physico-chemical properties, along with their growing commercialization and consumption, have made them ubiquitous in aquatic environments through both direct and indirect releases, raising significant concerns about their potential threats to inhabiting biota. Additionally, increasing surface water temperatures exacerbate ecological risks, making it imperative to understand the implications for non-target species at different biological levels. This study investigated the short- and long-term effects of UV-filters 4-MBC or BP-3, at ecologically relevant concentrations, combined with current and predicted warming scenarios, on the performance and male reproductive health of Mytilus galloprovincialis mussel populations. Using biomarkers across sub-cellular, cellular, tissue, and individual levels, the study revealed significant physiological and biochemical impairments in both sperm cells and adults exposed to UV-filters. Temperature emerged as the primary driver influencing mussel responses and modulating the impacts of 4-MBC/BP-3, emphasizing their sensitivity to temperatures outside the optimal range and interactive effects between stressors. Specifically, sperm motility declined with increasing UV-filter concentrations, while temperature alone influenced ROS production, leading to compromised mitochondrial activity and DNA damage in the presence of combined stressors, indicative of potential reproductive impairments. Adults exhibited high UV-filter bioconcentration potential in whole tissues, compromised physiological status, morphophysiological changes in digestive glands, oxidative stress, and alterations in metabolic capacity, antioxidant defences, and biotransformation mechanisms, correlating with UV-filter exposure and temperature increase. Among the UV-filters tested, 4-MBC was the most detrimental, especially when combined with warming. Overall, this study underscores the vulnerability of M. galloprovincialis to cumulative stressors and highlights the importance of employing a multi-biomarker approach to assess and mitigate the impacts of stressors on coastal ecosystems.

PON1 enzyme activity assays for serum and heparinized plasma in horses and stability evaluation of the enzyme activity over different freeze-thaw cycles and mimic transportation.

Consistent information and standardization procedures regarding the time of storage for frozen samples and the effects of storage time on enzyme activity are still missing in the literature. Thus, we evaluated the effects of different storage temperatures (-20 °C and - 80 °C), three repetitive freeze/thaw cycles, and 24-h mimic transportation on the activities of PON1 (paraoxonase and arylesterase), enzymes involved in the protection and detoxification processes of reactive molecules. PON1 enzymes' activity was validated on serum and heparinized plasma in horses. The results revealed that conditions and time of storage of blood samples for PON1 analyses altered the activities of both enzymes in both sample types, evidencing that these conditions can lead to protein degradation or general alteration. Specifically, paraoxonase and arylesterase activities significantly decreased among storage temperatures, with major effects detected at -20 °C. The repeated freeze/thaw cycles at -20 °C and 24-h mimic transport conditions also generated an expected degradation of the arylesterase in both serum and heparinized plasma while freeze/thaw cycles at -80 °C caused an increase of both arylesterase and paraoxonase activities on both sample types. In general, similar enzyme responses were detected between serum and heparinized plasma.

Mytilus galloprovincialis: A valuable bioindicator species for understanding the effects of diclofenac under warming conditions.

Drugs are chemical compounds used to treat and improve organic dysfunctions caused by diseases. These include analgesics, antibiotics, antidepressants, and antineoplastics. They can enter aquatic environments through wastewater streams, where their physico-chemical properties allow metabolites to distribute and accumulate. Current climate change and associated extreme weather events may significantly impact these substances' toxicity and aquatic organisms' sensitivity. Among the chemicals present in aquatic environments is the non-steroidal anti-inflammatory drug diclofenac (DIC), which the EU monitors due to its concentration levels. This study investigated the influence of temperature (control at 17 °C vs. 21 °C) on the effects of DIC (0 µg/L vs. 1 µg/L) in the mussel species Mytilus galloprovincialis. Significant results were observed between 17 and 21 °C. Organisms exposed to the higher temperature showed a decrease in several parameters, including metabolic capacity and detoxification, particularly with prolonged exposure. However, in some parameters, after 21 days, the M. galloprovincialis showed no differences from the control, indicating adaptation to the stress. The results of this study confirm that DIC concentrations in the environment, particularly when combined with increased temperatures, can produce oxidative stress and adversely affect M. galloprovincialis biochemical and physiological performance. This study also validates this species as a bioindicator for assessing environmental contamination with DIC. Beyond its direct impact on aquatic organisms, the presence of pharmaceuticals like DIC in the environment highlights the interconnectedness of human, animal, and ecosystem health, underscoring the One Health approach to understanding and mitigating environmental pollution.

The influence of temperature on the impacts of caffeine in mussels: Evaluating subcellular impacts and model predictions.

In aquatic ecosystems, the presence of pharmaceuticals, particularly caffeine (CAF), has been linked to wastewater discharge, hospital waste, and the disposal of expired pharmaceutical products containing CAF. Additionally, rising temperatures due to climate change are anticipated in aquatic environments. This study aimed to assess the toxicity of various CAF concentrations under current (17 °C) and projected (21 °C) temperature conditions, using the mussel Mytilus galloprovincialis as a bioindicator species. Subcellular impacts were evaluated following 28 days of exposure to four CAF concentrations (0.5; 1.0; 5.0; 10.0 µg/L) at the control temperature (17 °C). Only effects at an environmentally relevant CAF concentration (5.0 µg/L) were assessed at the highest temperature (21 °C). The overall biochemical response of mussels was evaluated using non-metric Multidimensional Scaling (MDS) and the Integrated Biomarker Response (IBR) index, while the Independent Action (IA) model was used to compare observed and predicted responses. Results showed that at 17 °C, increased CAF concentrations were associated with higher metabolism and biotransformation capacity, accompanied by cellular damage at the highest concentration. Conversely, under warming conditions (21 °C), the induction of antioxidant enzymes was observed, although insufficient to prevent cellular damage compared to the control temperature. Regarding neurotoxicity, at 17 °C, the activity of the acetylcholinesterase enzyme was inhibited up to 5.0 µg/L; however, at 10.0 µg/L, activity increased, possibly due to CAF competition for adenosine receptors. The IA model identified a synergistic response for most parameters when CAF and warming acted together, aligning with observed results, albeit with slightly lower magnitudes.

Six-Year Prescription Pattern of Antimicrobial Use in Cats at the Veterinary Teaching Hospital of the University of Pisa.

The use of antimicrobials has greatly contributed to improving animal health. However, their inappropriate use reduces their effectiveness in treating bacterial infections and contributes to the selection of resistance. This study aimed to retrospectively evaluate the six-year pattern (2017-2022) of antimicrobial use in cats visiting the Veterinary Teaching Hospital (VTH) of the University of Pisa (Italy). The total number of prescribed antimicrobials, the number of animals for which an antimicrobial was prescribed, and the total number of antimicrobial prescriptions showed a significant time trend decrease during the study period, except for the fixed-dose combinations. The most frequently prescribed antimicrobials were amoxicillin-clavulanic acid (Synulox) (39.1%) followed by enrofloxacin (29.8%). These antimicrobials were mostly prescribed to treat infections affecting the genitourinary tract (~30%), followed by the gastrointestinal tract, skin, and respiratory system affections. Antimicrobials in empirical associations represented 13.0% of the total antimicrobial prescriptions, and the combination of amoxicillin-clavulanic acid (Synulox) with enrofloxacin accounted for the majority. The oral route represented the main route of administration of prescribed antimicrobials, followed by parenteral and topical ones. Amoxicillin-clavulanic acid (Synulox) (37.2%), ceftriaxone (2.7%), and tobramycin (2.8%) were the most prescribed antimicrobials for the oral, parenteral, and topical routes, respectively. Antimicrobial prescriptions complied with prudent use recommendations in terms of availability of diagnosis, respect to the dose range, duration of treatment, and the use of medicinal products approved for the species. On the contrary, antimicrobial susceptibility tests were used infrequently (5.2%), lacking compliance with the existing guidelines observed in companion animal practice. Overall, additional interventions are required not only to improve the responsible use of antimicrobials in our feline practice but also to implement antimicrobial stewardship programs, enhancing diagnostics such as culture and sensitivity testing in the future.

Pattern of prescriptions and prudent use of antimicrobial in horse practice at a Veterinary Teaching Hospital.

Antimicrobial prescriptions of the University of Pisa and their compliance with prudent use recommendations were investigated over 11 years (2011-2021). At least one antimicrobial was always prescribed in surgical prophylaxis for the suture of wounds and in 33% of horses with signs of disease of a body system. Antimicrobials were administered in monotherapy (48%) in fixed dose combinations (21%) and empirical combinations (31%). Antimicrobials were mostly (63%) administered by parenteral route, while oral and topical antimicrobials accounted for 14% and 23% of prescriptions, respectively. Gentamicin, benzylpenicillin and ceftiofur were the most prescribed antimicrobials; aminoglycosides, penicillins and cephalosporins were the most common class of antimicrobial prescribed. Protected antimicrobials (WHO HPCIA and rifampicin) represented 24% of antimicrobial dispensations. The pattern of classes of antimicrobial used by body system was broad and included up to eight different pharmaceutical classes. The heterogeneity of antimicrobial use was confirmed by the estimate of the prescription diversity index. Antimicrobial prescriptions were in compliance with prudent use recommendations in terms of availability of diagnosis, respect of the dose range and duration of treatment On the contrary, principles of appropriate antimicrobial use have only been partially observed in relation to off-label use, use of antimicrobials in empirical combination, use of antimicrobial susceptibility tests and use of protected antimicrobials, suggesting that additional interventions are required to improve the responsible use of antimicrobials use in our equine practice.

Effect of amino acid enriched diets on hemolymph amino acid composition in honey bees.

Amino acids (AAs) are an abundant class of nectar solutes, and they are involved in the nectar attractiveness to flower visitors. Among the various AAs, proline is the most abundant proteogenic AA, and γ-amino butyric acid (GABA) and ß-alanine are the two most abundant non-proteogenic AAs. These three AAs are known to affect insect physiology, being involved in flight metabolism and neurotransmission. The aim of this study was to investigate the effects of artificial diets enriched with either ß-alanine, GABA, or proline on consumption, survival, and hemolymph composition in honey bees belonging to two different ages and with different metabolism (i.e., newly emerged and foragers). Differences in feed intake among diets were not observed, while a diet enriched with ß-alanine improved the survival rate of newly emerged honey bees compared to the control group. Variations in the hemolymph AA concentrations occurred only in newly emerged honey bees, according to the diet and the time of hemolymph sampling. A greater susceptibility of young honey bees to enriched diets than older honey bees was observed. The variations in the concentrations of hemolymph AAs reflect either the accumulation of dietary AAs or the existence of metabolic pathways that may lead to the conversion of dietary AAs into different ones. This investigation could be an initial contribution to studying the complex dynamics that regulate hemolymph AA composition and its effect on honey bee physiology.

Exploring toxicological interactions in a changing sea: The case of the alkaloids caffeine and caulerpin.

The bisindolic alkaloid caulerpin (CAU) is a bioactive compound isolated from green algae of the genus Caulerpa that are highly invasive in the Mediterranean Sea. On the other side, the purine alkaloid caffeine (CAF) is one of the most globally consumed psychoactive substances and a widespread anthropogenic water pollutant. Both compounds display a large panel of biological properties and are well known to accumulate in the tissues of aquatic organisms and, in certain circumstances, co-occur in the human diet. On this premise, the present study aimed to investigate possible synergistic interactions between CAU and CAF by using the bivalve Mytilus galloprovincialis as a model organism. Mussels were exposed to CAF via medium while they were fed with food enriched with CAU. After treatments, biochemical analysis confirmed the toxic potential of CAF, with increased AChE activity and lipid peroxidation. Also, histopathological alterations were observed in the gills and digestive tubules. The NMR-based metabolomics analysis detected higher levels of free amino acids under CAF treatments. Conversely, the food administration of CAU did not affect the above toxicological biomarkers. In addition, we did not observe any cumulative effect between CAF and CAU toward increased cellular damage and neurotoxicity. On the other hand, a possible action of CAU in decreasing CAF toxicity could be hypothesized based on our results. This hypothesis is supported by the activity of CAU as an agonist of peroxisome proliferator-activated receptors (PPARs). PPARs mediate xenobiotic detoxification via cytochromes P450, which is involved in CAF metabolism. Overall, the results obtained not only rule out any cumulative adverse effects of CAF and CAU but also encourage further research to evaluate the possible use of CAU, a compound easily obtained through the valorization of biomass from invasive species, as a food additive to improve the clearance of xenobiotics.

Procalcitonin and carbonylated protein concentrations in equine synovial fluid.

Early diagnosis of joint diseases is fundamental for prompt and appropriate management, particularly in septic arthritis. Procalcitonin (PCT) and protein carbonylated content (PCC) have been investigated in both human and veterinary medicine. An increase in PCT has been shown in infectious bacterial diseases, while higher levels of PCC have been shown in inflammatory pathologies characterized by oxidative damage. This study evaluated PCT and PCC in plasma and synovial fluid (SF), in healthy and pathological equine joints, affected by different types of arthropathy. Twenty-nine joints were evaluated and underwent orthopedic, radiographic, ultrasonographic and SF evaluation. The joints were divided in three groups: healthy, septic, and non-septic arthritis. PCT and PCC were measured in horse plasma and SF. Data distribution was evaluated and results were expressed as median, quartile values. Statistical differences in SF values among groups and correlations were assessed between plasma and SF of both PCT and PCC. The groups of joint disease included: 8/29 healthy, 13/29 non-septic and 8/29 septic. Significant differences were obtained for SF PCC and plasma PCT between healthy and septic joints, while no differences were found for plasma PCC and SF PCT. A positive correlation was found between plasma and SF PCT. To the best of our knowledge, this is the first study reporting PCT in equine SF. SF PCC could be a useful biomarker to differentiate between septic and healthy joints.

Case-Control Study: Endogenous Procalcitonin and Protein Carbonylated Content as a Potential Biomarker of Subclinical Mastitis in Dairy Cows.

Procalcitonin (PCT) and protein carbonylated content (PCC) are promising biomarkers for bacterial infection and inflammation in veterinary medicine. This study examined plasma PCT and PCC levels in healthy cows (H) and cows with subclinical mastitis (SCM). A total of 130 cows (65 H and 65 SCM) were included in this study. Blood samples were collected, and plasma was frozen at -80 °C. PCT levels were determined using a bovine procalcitonin ELISA kit, while PCC was measured following the methodology of Levine et al. Statistical analysis revealed a significant difference in PCT levels between H (75.4 pg/mL) and SCM (107.3 pg/mL) cows (p < 0.001) and significantly lower concentrations of PCC in the SCM group (H: 0.102 nmol/mL/mg, SCM: 0.046 nmol/mL/mg; p < 0.001). The PCT cut-off value for distinguishing healthy and subclinical mastitis animals was >89.8 pg/mL (AUC 0.695), with a sensitivity of 66.2% and specificity of 69.2%. PCT showed potential value as a diagnostic tool to help in decision making for subclinical mastitis cases, while PCC requires further studies to investigate the trend of this biomarker during localized pathology.

Multi-biomarker approach for the (eco)toxicity of UV-filter environmental pollution on the Mediterranean mussel Mytilus galloprovincialis in a multiple stressor context. The case of 4-MBC under salinity shifts.

Marine-coastal ecosystems are rapidly transforming because of climate change (CC). At the same time, the impacts of emerging organic contaminants (i.e., organic UV-filters) on these ecosystems are intensifying. In the Mediterranean, the consequences of these disturbances are occurring at a fast pace making this area a potential sentinel site to be investigated. While singular effects of organic UV-filters or CC-related factors on marine biota have been relatively described, their combined impact is still largely unknown. Thus, the objective of this study was to assess the long-term responses of the Mediterranean mussel Mytilus galloprovincialis towards anticipated salinity changes (decreases-S20 or increases-S40) when exposed to environmentally relevant concentrations of the UV-filter 4-methylbenzylidene camphor (4-MBC). An integrated multi-biomarker approach was applied, featuring general and oxidative stress, antioxidant and biotransformation enzyme capacity, energy metabolism, genotoxicity, and neurotoxicity biomarkers. Results showed that both projected salinities, considered separately, exerted non-negligible impacts on mussels' health status, with greater biological impairments found at S 40. Combining both stressors resulted in an evident increase in mussels' susceptibility to the UV-filter, which exacerbated the toxicity of 4-MBC. The dominant influence of salinity in the climate change-contaminant interaction played a crucial role in this outcome. The most severe scenario occurred when S 20 was combined with 4-MBC. In this situation, mussels exhibited a decrease in filtration rate, metabolic capacity and deployment of energy reserves increased, with an upregulation of biotransformation and inhibition of antioxidant enzyme activities. This exposure also led to the observation of cellular and DNA damage, as well as an increase in AChE activity. Furthermore, salinity-dependent bioaccumulation patterns were evaluated revealing that the lowest values in contaminated mussels are found at S 20. Overall, the present findings provide evidence that projected CC/pollutant scenarios may represent high risks for mussels' populations, with global relevant implications for the ecosystem level.

Oxidative Status and Histological Evaluation of Wild Boars' Tissues Positive for Zearalenone Contamination in the Campania Region, Southern Italy.

Zearalenone (ZEN) is a mycotoxin produced by fungi belonging to the genera Fusarium spp. and commonly found in feed and food. It is frequently related to reproductive disorders in farm animals and, occasionally, to hyperestrogenic syndromes in humans. Nowadays, knowledge about ZEN effects on wild boars (Sus scrofa) is extremely scarce, despite the fact that they represent one of the most hunted game species in Italy. The aim of this study was to investigate how ZEN affects the liver, kidney, and muscle oxidative status and morphology of wild boars hunted in various locations throughout the province of Avellino, Campania Region, Southern Italy, during the 2021-2022 hunting season. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activities, as well as the malondialdehyde (MDA) levels, were assessed by colorimetric assays; tissue morphology was evaluated by hematoxylin-eosin and Masson's stains. Our data showed that ZEN contamination might result in oxidative stress (OS) and some histopathological alterations in wild boars' livers and kidneys rather than in muscles, emphasizing the importance of developing a wildlife monitoring and management strategy for dealing not only with the problem of ZEN but the surveillance of mycotoxins in general.

Sub-lethal effects induced in Mytilus galloprovincialis after short-term exposure to sodium lauryl sulfate: Comparison of the biological responses given by mussels under two temperature scenarios.

Surfactants are among the most common PPCPs that reach coastal systems, being often used in large quantities in cleaning products such as detergents and soap powders. Sodium lauryl sulfate (SLS) is listed in this group of emerging contaminants. Previous studies have already demonstrated the presence of SLS in aquatic environments and the negative effects on organisms living there. However, with ocean acidification and warming predictions, SLS-induced impacts may differ from those currently known. In this context, the present study aimed to reproduce environmental conditions by assessing the release of substances over a short period and to understand the influence of a rapid increase in temperature on the impacts caused. The marine bivalve Mytilus galloprovincialis was exposed to 2.0 mg/L SLS at 17 °C and 21 °C for 7 days. To assess the possible biochemical changes resulting from the exposure of mussels to SLS, a series of biomarkers related to oxidative stress/damage, detoxification, and metabolic capacity were measured. The SLS accumulation in soft tissues was low (about 0.7 ng/g) at both temperatures. The results evidenced increased metabolic activity, especially in mussels exposed to SLS at 17 °C. An increase in protein content was also observed upon exposure to SLS and increased temperature compared to controls at 17 °C. Although no effects on antioxidant enzymes were observed, protein damage was recorded, especially at 21 °C. These findings confirmed that SLS induces toxic effects and predicted climate change factors may increase the impact on M. galloprovincialis.

Combined effects of temperature rise and sodium lauryl sulfate in the Mediterranean mussel.

Personal care products (PCPs) are those compounds used daily (e.g., soaps, shampoos, deodorants, and toothpaste), explaining their frequent detection in aquatic systems. Still, scarce information is available on their effects on inhabiting wildlife. Among the most commonly used PCPs is the surfactant Sodium Lauryl Sulfate (SLS). The present study investigated the influence of temperature (CTL 17 ºC vs 22 ºC) on the effects of SLS (0 mg/L vs 4 mg/L) in the mussel species Mytilus galloprovincialis. Mussels' general health status was investigated, assessing their metabolic and oxidative stress responses. Higher biochemical alterations were observed in SLS-exposed mussels and warming enhanced the impacts, namely in terms of biotransformation capacity and loss of redox homeostasis, which may result in consequences to population maintenance, especially if under additional environmental stressors. These results confirm M. galloprovincialis as an excellent bioindicator of PCPs pollution, and the need to consider actual and predicted climate changes.

A forecast effects of climate change and anthropogenic compounds in Gambusia holbrooki: ecotoxicological effects of salinity and metformin.

Due to global warming and extreme weather events, estuarine and coastal ecosystems are facing sudden fluctuations in salinity. These ecosystems are also threatened by organic and inorganic compounds that increase water pollution. Metformin is an antidiabetic drug commonly used by patients with type-2 diabetes, and an increase in environmental concentration has been recorded. To better understand the impacts of these two stressors on aquatic organisms, this study assessed: 1) the acute (96 h) ecotoxicological effects (antioxidant and biotransformation capacity, oxidative damage, energetic reserves, and protein content, neurotoxicity) induced by a range of metformin concentrations in Gambusia holbrooki under different salinities (17, 24, 31 expressed as Practical Salinity Units - PSU); and 2) the same endpoints after chronic exposure (28 d) under a range of metformin concentrations at a salinity of 17. The results obtained from the acute exposure showed interactions between salinity and metformin in G. holbrooki superoxide dismutase (SOD) activity, body protein, and glycogen (GLY) contents. The results revealed that an increase in salinity can modulate the response of G. holbrooki to metformin. Chronically exposed organisms showed that metformin led to a significant decrease in SOD activity at most of the tested concentrations (0.5, 1.0, and 10 µg/L). In addition, glutathione S-transferases increased and glutathione peroxidase activity decreased significantly at concentrations of metformin of 5 and 10 at the µg/L, respectively. Therefore, overall, metformin can lead to potential oxidative stress in G. holbrooki the highest metformin concentrations tested and the GLY content in G. holbrooki increased after exposure to metformin concentrations of 0.5, 1.0 and 5.0 µg/L. Published studies have already shown that metformin alone can lead to oxidative damage in aquatic species, endangering the biodiversity of aquatic ecosystems. Therefore, additional ecotoxicological studies should be performed to characterize if other metformin concentrations combined with salinity, or other climate change-related factors, might impact non-target species. Standard toxicity bioassays may not be predictive of actual pollutants (e.g. metformin) toxicity under variable environmental conditions, and the investigation of a wider range of exposure conditions could improve the accuracy of chemical risk assessments.

Will warmer summers increase the impact of UV filters on marine bivalves?

Marine organisms are constantly exposed to multiple stressors including pollutants released into the environment, such as personal care products (PCPs), and climate change-derived factors, namely warming, which are aggravated by anthropogenic drivers and pose increasing pressure on coastal ecosystems. Avobenzone (AVO) is one of the most used ultraviolet (UV) filters in PCPs which have been increasingly used and, thereby, identified in aquatic environments. However, data regarding the influence of warming on the impacts caused by AVO in bivalves is lacking. Mussels are considered good bioindicators thus being often employed in ecotoxicology studies. Hence, the present study aimed to evaluate the toxic effects of an environmentally relevant concentration of AVO (0.5 µg/L) and warming (21 °C), acting alone or in combination, on sperm and adults of the Mediterranean mussel species Mytilus galloprovincialis, through in vitro and in vivo tests, respectively. AVO and warming effects were evaluated by assessing oxidative status, viability, genotoxicity, motility, and kinetics in sperm, together with the quantification of energy content, metabolic capacity, biological defence mechanisms, cellular damage, and neurotoxicity in adults. AVO induced genotoxicity and increased respiration rate in sperm while enhancing the biotransformation enzymes' activity in adults. Exposure to warming led to an increase in respiration rate, ROS overproduction, cellular damage, and viability decrease in sperm whereas metabolic capacity increased in adults. AVO combined with warming caused oxidative stress, cellular damage, genotoxicity, and decreased motility in sperm, while only antioxidant enzymes' activity was enhanced in adults. Overall, the present study demonstrated that when acting in combination the effects of both stressors were more prominent. Furthermore, considering the multiple-stressor scenario tested, major toxic effects occurred in male gametes in comparison to adults.

Zearalenone (ZEN) and Its Metabolite Levels in Tissues of Wild Boar (Sus scrofa) from Southern Italy: A Pilot Study.

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by the fungi of the Fusarium genera, and is a contaminant of cereals and plant products. ZEN and its metabolites are considered endocrine disruptors, and could have various toxic effects on animals and humans. In recent years, there has been a significant demographic increase in wild boar (Sus scrofa) in many mountainous and hilly areas of Italy, including the Campania region, mainly due to global climate change. The wild boar can be defined as a generalist and omnivorous species capable of varying its diet; therefore, it can play a role as an environmental bioindicator towards contaminants such as mycotoxins. This study was conducted to evaluate, for the first time, the concentrations of ZEN and its metabolites in the liver, kidney, and muscle of 82 wild boars shot in their habitat by hunters with hunting permits in different localities of Avellino province (Campania region, Southern Italy) from 2021 to 2022. The samples were collected and analyzed with an SPE clean-up and high-pressure liquid chromatography method with fluorescence detection. The results indicated that ZEN and α-Zearalenol were present in most of the samples, suggesting that a plan to monitor these mycoestrogens is essential to achieve the goals of "One Health".

Toxic effects of a mixture of pharmaceuticals in Mytilus galloprovincialis: The case of 17α-ethinylestradiol and salicylic acid.

The impact of pharmaceuticals on marine invertebrates has been a topic of rising concern, with an increasing number of studies regarding the impacts on bivalves. However, very few investigated the toxicity of mixtures of pharmaceuticals. This knowledge gap was investigated in the present study, where the toxicity of 17α-ethinylestradiol (EE2) and salicylic acid (SA) mixture was evaluated. To this end, Mytilus galloprovincialis mussels were chronically subjected to both pharmaceuticals, acting alone and in combination, and the effects at the cellular level were measured. The Independent Action (IA) model was performed aiming to compare obtained with predicted responses. The integrated biomarker response (IBR) index was used to assess the overall biochemical response given by mussels. The results obtained revealed that the most stressful condition was caused by the combined effect of EE2 and SA, with the highest metabolic capacity, antioxidant (catalase activity) and biotransformation (carboxylesterases activity) activation and cellular damage in organisms exposed to the mixture of both drugs in comparison to responses observed when each drug was acting alone. Predicted responses obtained from the IA model indicate that caution should be paid as frequent deviations to observed responses were found. This study highlights the need for future studies considering the mixture of pollutants, mimicking the actual environmental conditions.