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".

Case - Control study: Evaluation of plasma procalcitonin concentration as an indicator of inflammation in healthy and sick cows.

This case - control study aims to evaluate Procalcitonin (PCT) plasma concentrations in healthy and hospitalized cows with a conclusive diagnosis of inflammation due to bacterial infection. Thirty-four healthy and 131 sick cows were included. Procalcitonin concentrations were assessed using an ELISA kit for cattle. Depending on whether sick cows received antimicrobial treatments prior to admission or not, they were divided in treated (TP) or not treated (NTP) subgroups. Mann-Whitney U tests were performed to determine differences between healthy vs sick cows, while Kruskal-Wallis with Dunn's multiple comparison test were applied for healthy vs sick subgroups. Receiver operating characteristic (ROC) analysis was performed to assess the optimal cut-off value. Kaplan-Meier survival curves were determined for cows belonging to the groups with PCT values below and above ROC cut-offs. Plasma PCT concentration was 200.1 (147.8-324.1) pg/mL and 361.6 (239.7-947.1) pg/mL in the healthy control and in the sick group, respectively (P < 0.001). The optimal cut-off value of plasma PCT concentration was 244.4 pg/mL (sensitivity 73.6%, specificity 60.0%). The plasma PCT concentration was 267.5 (210.3-771.2) pg/mL in the TP subgroup and 425.6 (253.1-1242) pg/mL in the NTP subgroup (P = 0.03). Cows with PCT above the ROC cut-off value had a reduced survival percentage and a higher mortality risk (P < 0.05). Procalcitonin showed the ability of differentiate healthy cows from hospitalized cows with a conclusive diagnosis of inflammation due to bacterial infection. Moreover, PCT was a good predictor of negative prognostic outcome.

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.

Gene expression and biochemical patterns in the digestive gland of the mussel Mytilus galloprovincialis (Lamarck, 1819) exposed to 17α-ethinylestradiol.

Contaminants of emerging concern (CECs) are a class of chemicals that can spread throughout the environment and may cause adverse biological and ecological effects. While there are many different classes of CECs, one of the most well documented in the aquatic environment are pharmaceutical drugs, such as natural and synthetic estrogens. In particular, the widespread presence of the synthetic estrogen 17 α-Ethinylestradiol (EE2) in water may lead to bioaccumulation in sediment and biota. EE2 is the primary component in contraceptive pills, and is a derivative of the natural hormone estradiol (E2). In this study, the mussel Mytilus galloprovincialis was exposed to EE2 in a semi-static and time-dependent experiment, for a total exposure period of 28 days. Biochemical and transcriptomics analyses were performed on mussel digestive glands after exposure for 14 (T14) and 28 (T28) days. Metabolic and DNA impairments, as well as activation of antioxidant and biotransformation enzymes activation, were detected in T28 exposed mussels. RNA-Seq analysis showed significant differential expression of 160 (T14 compared to controls), 33 (T28 compared to controls) and 79 (T14 compared to T28) genes. Signs of stress after EE2 treatment included up-regulation of gene/proteins involved with immune function, lipid transport, and metabolic and antibacterial properties. This study elucidates the underlying mechanisms of EE2 in a filter feeding organisms to elucidate the effects of this human pharmaceutical on aquatic biota.

Molecularly imprinted polymers as effective capturing receptors in a pseudo-ELISA immunoassay for procalcitonin detection in veterinary species.

In this study, a new sandwich-type immunoenzymatic assay, based on a molecularly imprinted polymer (MIP) as an artificial antibody (pseudo-ELISA), was developed for the determination of procalcitonin (PCT) in veterinary species. The quantification of PCT in human medicine represents the state of the art for the diagnosis of sepsis; instead the clinical studies on the relevance of PCT as a sepsis predictor in veterinary patients are few, likely due to the total absence of validated assays. MIPs have been widely used as antibody mimics for important applications, and MIP-based sandwich assays have emerged as promising analytical tools for the detection of disease biomarkers. Herein, a polynorepinephrine (PNE)-based imprinted film was directly synthesized on the well surface of a 96-well plate. Subsequently, based on a commercial ELISA kit, the PCT quantification was accomplished via a colorimetric sandwich assay by replacing the capture antibody of the kit with the PNE-based MIP. This method was performed to detect canine and equine PCT in buffer and in plasma samples. Under optimal conditions, the results obtained in plasma samples showed a limit of detection (LOD) of 5.87 ng mL-1 and a reproducibility (CVav%) of 10.0% for canine samples, while a LOD = 4.46 ng mL-1 and CVav% = 7.61% were obtained for equine samples.

In vitro spermiotoxicity and in vivo adults' biochemical pattern after exposure of the Mediterranean mussel to the sunscreen avobenzone.

Avobenzone (AVO) is one of the most frequent ultraviolet (UV) filters in personal care products (PCPs). The Mediterranean mussel Mytilus galloprovincialis is a bioindicator often used for ecotoxicological research. Since UV filters reach higher peaks during summer in aquatic bodies, coincident with mussels' spawning period, and bivalves are sessile, both male gametes and adults of this species were used in this experiment. Therefore, the present study aimed to assess how AVO affects M. galloprovincialis at different biological levels. In vitro experiments on sperms (30 min-exposure) and in vivo experiments on adults (28 days-exposure) were carried out at 0.1, 1.0 and 10.0 µg/L of AVO concentrations. The oxidative and physiological status together with genotoxicity in exposed sperms were assessed. Several biochemical parameters related to enzymatic antioxidant defences, biotransformation enzymes, cell membrane damage, energy reserves, and neurotoxicity were evaluated in adult mussels. Results of in vitro sperm exposure to AVO showed significant overproduction of superoxide anions and DNA damages in all treatments and decrease in sperm viability at 1.0 and 10.0 µg/L. AVO exposure also led to complete inhibition of motility of sperms at the highest concentration, while a significant increase of curvilinear velocity and decrease of wobble occurred at 1.0 µg/L. In vivo exposed adults exhibited a significant decrease in metabolic capacity at 0.1 µg/L, a significant increase in the total protein content and enzymatic turnover as superoxide dismutase (antioxidant defence) at 10 µg/L. This study revealed an ecological concern related to the high sensitivity of sperms respectively to adults under environmentally relevant concentrations of AVO, underpinning an hypothesis of male reproductive function impairments.

Prevalence, clinical features, and outcome of intervertebral disc extrusion associated with extensive epidural hemorrhage in a population of French Bulldogs compared to Dachshunds.

Intervertebral disc extrusion associated with extensive epidural hemorrhage (DEEH) is a well-documented pathological condition in veterinary medicine. This retrospective study aimed to evaluate the prevalence and clinical features of DEEH in a population of French Bulldogs affected by intervertebral disc extrusion (n=75), compare the findings with those from a group of Dachshunds (n=98) and identify possible predictive factors of DEEH and outcomes in surgically treated patients. The study showed that the prevalence of DEEH observed in Dachshunds (11.2% [95% confidence interval [CI]: 5.7-19.2%]) was significantly lower than that observed in French Bulldogs (41.3% [95% CI: 30.1-53.3%]). The multiple logistic regression model highlighted that the patients presenting with an acute onset of clinical signs (>24 hr) (odds ratio [OR]: 13.08; 95% CI: 4.63-37.03, P=0.00), presence of clinical signs progression (OR: 5.04; P=0.01), and French Bulldogs (OR: 5.15; 95% CI: 1.71-15.54, P=0.00) were at increased risk of developing DEEH. Secondary analysis showed that patients with DEEH were at an increased risk of being non-ambulatory at discharge (OR: 3.43; P=0.017). Overall, the surgically treated patients had favorable outcomes.

Ecotoxicological effects of the UV-filter 4-MBC on sperms and adults of the mussel Mytilus galloprovincialis.

Present in an increasing number of products, UV-filters are continuously discharged into aquatic environments. Despite potential risks for inhabiting organisms are recognized, the effects of UV-filter 4-methylbenzylidenecamphor (4-MBC) on marine invertebrates are poorly investigated. By combining in vitro/in vivo exposures through a multi-biomarker approach on sperms and adults, the present study evaluated how 4-MBC affect the mussel species Mytilus galloprovincialis, providing ecologically relevant information on organisms' responses. From the obtained results, considering mortality as endpoint, sperms revealed a greater sensitivity (EC50:347 µg/L) than adults (EC50: not calculable). From an ecotoxicological perspective, this resulted in a derived threshold concentration (LOEC) of 100 µg/L and 72 µg/L, respectively. Effects at the cell/molecular level were provided by general redox-status imbalance and oxidative stress. Sperms showed functional and structural impairments, hyperactivation and DNA damage, while adults showed physiological, metabolic/energetic dysfunctions, DNA damage and activation of oxidative and biotransformation enzymes. High 4-MBC bioaccumulation was also observed in exposed mussels (BCFs:14.0-32.0 L/kg). These findings suggest that 4-MBC may impair fitness and survival of the broadcast spawning mussel M. galloprovincialis, affecting reproduction success and population growth.