Toxic impacts induced by Sodium lauryl sulfate in Mytilus galloprovincialis.

Pharmaceuticals and personal care products (PPCPs) are continuously dispersed into the environment, as a result of human and veterinary use, reaching aquatic coastal systems and inhabiting organisms. However, information regarding to toxic effects of these compounds towards marine invertebrates is still scarce, especially in what regards to metabolic capacity and oxidative status alterations induced in bivalves after chronic exposure. In the present study, the toxic impacts of Sodium lauryl sulfate (SLS), an anionic surfactant widely used as an emulsifying cleaning agent in household and cosmetics, were evaluated in the mussel Mytilus galloprovincialis, after exposure for 28 days to different concentrations (0.0; 0.5; 1.0; 2.0 and 4.0 mg/L). For this, effects on mussels respitation rate, metabolic capacity and oxidative status were evaluated. The obtained results indicate a significant decrease on mussel's respiration rate after exposure to different SLS concentrations, an alteration that was accompanied by a decrease of bioconcentration factor along the increasing exposure gradient, especially at the highest exposure concentration. Nonetheless, the amount of SLS accumulated in organisms originated alterations in mussel's metabolic performance, with higher metabolic capacity up to 2.0 mg/L followed by a decrease at the highest tested concentration (4.0 mg/L). Mussels exposed to SLS revealed limited antioxidant defense mecanhisms but cellular damage was only observed at the highest exposure concentration (4.0 mg/L). In fact, up to 2.0 mg/L of SLS limited toxic impacts were observed, namely in terms of oxidative stress and redox balance. However, since mussel's respiration rate was greatly affected by the presence of SLS, the present study may highlight the potential threat of SLS towards marine bivalves, limiting their filtration capacity and, thus, affecting their global physiological development (including growth and reproduction) and ultimely their biochemical performance (afecting their defense capacity towards stressful conditons).

The influence of climate change related factors on the response of two clam species to diclofenac.

Diclofenac (DIC) is one of the non-steroidal anti-inflammatory drugs (NSAID) with higher consumption rates, used in both human and veterinary medicine. Previous studies already demonstrated the presence of this drug in aquatic environments and adverse effects towards inhabiting organisms. However, with the predictions of ocean acidification and warming, the impacts induced by DIC may differ from what is presently known and can be species-dependent. Thus, the present study aimed to comparatively assess the effects caused by DIC in the clams Ruditapes philippinarum and Ruditapes decussatus and evaluate if these impacts were influenced by pH and temperature. For this, organisms were acclimated for 30 days at two different temperature and pH (control conditions: pH 8.1, 17 °C; climate change forecasted scenario: pH 7.7, 20 °C) in the absence of drugs (experimental period I) followed by 7 days exposure under the same water physical parameters but in absence or presence of the pharmaceutical drug (at 1 µg/L, experimental period II). Biochemical responses covering metabolic capacity, oxidative stress and damage-related biomarkers were contrasted in clams at the end of the second experimental period. The results showed that under actual conditions, R. philippinarum individuals exposed to DIC presented enhanced antioxidant activities and reduced their respiration rate compared with non-contaminated clams. When exposed to the predicted climate change conditions, a similar response was observed in contaminated clams, but in this case clams increased their metabolic activities probably to fight the stress caused by the combination of both stressors. When R. decussatus was exposed to DIC, even at actual pH and temperature conditions, their antioxidant defences were also elevated but their baseline enzymatic activities were also naturally higher in respect to R. philippinarum. Although clams may use different strategies to prevent DIC damage, both clam species showed under low pH and high temperature limited oxidative stress impacts in line with a lower DIC bioaccumulation. The present findings reveal that predicted climate change related factors may not enhance the impacts of DIC in Ruditapes clams in a species-dependent manner although both displayed particular mechanisms to face stress.

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.

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.

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.

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".

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.

Use of antimicrobials in companion animal practice: a retrospective study in a veterinary teaching hospital in Italy.

OBJECTIVES: To describe the use of antimicrobials in a veterinary teaching hospital for companion animals in Italy, with particular regard to the agreement with recommendations of prudent use METHODS: The study was conducted with a retrospective, cross-sectional design. The population under investigation included 18,905 cats and dogs that were referred to the hospital between 2000 and 2007. Two different samples of the clinical paper forms were randomly selected to estimate the prevalence of animals receiving an antimicrobial prescription and to describe the pattern of antimicrobials used in relation to the condition being treated. The proportion of antimicrobials prescribed accomplishing recommendations of prudent use was also estimated, as well as the level of agreement with specific, diagnosis-based guidelines for antimicrobial use. RESULTS: Broad-spectrum antimicrobials, including penicillins with ß-lactamase inhibitors, first-generation cephalosporins and fluoroquinolones, were the most frequently prescribed compounds. Antimicrobials prescribed with the support of microbiological analyses and susceptibility testing were less than 5%. Among the recommendation of prudent use, the availability of information from laboratory testing had the poorest degree of agreement, while the other evaluated items were accomplished in most of the cases. CONCLUSIONS: Our results highlight the need to improve the procedures of antimicrobial prescription in the study setting. This can be achieved by supporting the guidance for antimicrobial use at the local level, with the adoption of specific guidelines, and at the national level with a further implementation of the policies of prudent prescriptions.

Rapid and simultaneous electrochemical method to measure copper and lead in canine liver biopsy.

The conventional analytical techniques used for the quantitative analysis of heavy metals in animal tissues are the atomic absorption spectroscopy (AAS) and the inductively-coupled plasma mass spectrometry (ICP-MS). These methods involve high cost, skilled personnel and long analysis times. Several studies have shown the applicability of electrochemical transducer coupled to disposable screen-printed electrodes (SPE) for measuring metals. The aim of the present study was to applicate and validate a simple and fast protocol for the simultaneous measurement of Cu and Pb concentrations in canine liver biopsy, coupling a simple digestion procedure with electrochemical stripping analysis. Square wave anodic stripping voltammetry (SWASV) coupled to disposable SPE was employed as fast and sensitive method for metals detection. Samples were digested with hydrogen peroxide/hydrochloric acid mixture for 1 h coupled to solid phase purification with carbon columns. Instrumental precision, digestion procedure accuracy and limits of detections were evaluated. To validate the proposed method the metal concentrations have also been determined by AAS. Instrumental precision and digestion method accuracy were less than 15.3% and 16.8% for Cu and Pb, respectively. The limits of detection were 10 and 6 mg/kg dry weight in liver samples for Cu and Pb, respectively. The results obtained by the electrochemical method are in agreement with those of the reference method. The obtained results showed a reliable approach for Cu and Pb detection simultaneously with good sensitivity, accuracy and precision. The simple sample pre-treatment together with the low cost makes this approach particularly appealing for Cu and Pb quantification for diagnostic purposes.•Simultaneous determination of Cu and Pb concentrations in canine liver biopsy.•Square wave anodic stripping voltammetry (SWASV) with screen-printed electrodes (SPE) coupled to an acid digestion protocol.•Cu and Pb were measured with a very good sensitivity and accuracy.•Good agreement with AAS reference method.•Low cost, fast and sensitive method for diagnostic purposes.

A prospective multicenter observational study of Viperidae polyvalent immune F(ab')2 antivenom administration for the treatment of viper envenomation in dogs.

OBJECTIVE: To test an equine-derived polyvalent viperid antivenom (EPVA) in the treatment of dogs with evidence of viper envenomation. DESIGN: Prospective, multicenter observational study. SETTING: Veterinary emergency and critical care hospitals. ANIMALS: A total of 82 client-owned dogs with progressive clinical signs after viperid snakebite were enrolled in the study. INTERVENTIONS: Equine-derived polyvalent viperid antivenom was administered at a dosage of 1 mL/kg body weight, either by IV infusion or SC injection. MEASUREMENTS AND MAIN RESULTS: A standardized snakebite severity score (SSS) was used to characterize the severity of envenomation and the clinical course after EPVA treatment. Most dogs had improved SSS both at 4 (65.8%) and 8 hours (81.7%) following EPVA administration. Five dogs died. At the 4-week assessment, 3 dogs had slightly abnormal hematological or coagulation parameters; all other surviving dogs showed no abnormalities. Antivenom-related acute or intermediate reactions occurred in 12 dogs (14.6%). CONCLUSIONS: In the first study on antivenom in dogs in Italy, the effects of progressive viper envenomation were stabilized or reversed in the large majority of dogs receiving EPVA, as confirmed by the SSS analyses.

Novel organ-specific effects of Ketoprofen and its enantiomer, dexketoprofen on toxicological response transcripts and their functional products in salmon.

Racemic ketoprofen (RS-KP) and its enantiomer, dexketoprofen (S(+)-KP) are widely used non-steroidal anti-inflammatory drugs (NSAIDs), and commonly detected in the aquatic environment. The present study has evaluated the toxicological effects of RS-KP and S(+)-KP on biotransformation and oxidative stress responses in gills and liver of Atlantic salmon. Fish were exposed for 10 days using different concentrations of RS-KP (1, 10 and 100 µg/L) and S(+)-KP (0.5, 5 and 50 µg/L). Biotransformation and oxidative stress responses were analysed at both transcript and functional levels. In the gills, significant inhibitory effect at transcriptional and enzymatic levels were observed for biotransformation and oxidative stress responses. On the contrary, biotransformation responses were significantly increased at transcriptional and translational levels in the liver, while the associated enzymatic activities did not parallel this trend and were inhibited and further demonstrated by principal component analysis (PCA). Our findings showed that both compounds produced comparable toxicological effects, by producing organ-specific effect differences. RS-KP and S(+)-KP did not bioaccumulate in fish muscle, either due to rapid metabolism or excretion as a result of their hydrophobic properties. Interestingly, the inhibitory effects observed in the gills suggest that these drugs may not undergo first pass metabolism, that might result to downstream differences in toxicological outcomes.

Procalcitonin Detection in Veterinary Species: Investigation of Commercial ELISA Kits.

In human medicine, procalcitonin (PCT), the precursor of calcitonin, is used for the rapid identification of the origin and severity of sepsis. In veterinary medicine, PCT has been studied in horses, cattle, and dogs, but the use of PCT in diagnostic and/or prognostic settings is not possible because of the lack of validated assays to obtain reference ranges. The aim of the present study was the investigation of commercially available ELISA kits for the detection of canine and equine PCT in plasma samples. Validation of the ELISA kits was performed by using species-specific recombinant proteins spiked both in plasma and buffer samples; linearity, limit of detection (LOD), recovery, and intra-assay and inter-assay variability were calculated. Moreover, clinical samples obtained from sick and healthy animals were also analyzed with the tested kits. Canine PCT was measured with a recombinant canine and a canine PCT ELISA kit. Equine PCT was measured with an equine and a human ELISA PCT kit. Our data demonstrate that the canine recombinant PCT ELISA kit can be used to measure canine PCT in plasma samples, showing an intra-assay and inter-assay coefficient of variation less than 20% and a LOD of 11 pg/mL, whereas the present results do not support the use of the canine PCT ELISA kit. The human PCT ELISA kit is suitable to detect equine PCT with a LOD of 56 ng/mL, whereas the equine PCT ELISA kit did not detect recombinant equine PCT.

Evaluation of Protein Carbonyl Content in Healthy and Sick Hospitalized Horses.

Literature on the protein carbonyl content (PCC) evaluation in horses is scarce, thus the aims were to evaluate the PCC in healthy and SIRS (Systemic Inflammatory Response Syndrome) horses and to investigate the performances of PCC in terms of sensitivity, specificity, and likelihood ratio in identifying SIRS positive and negative horses. A total of 72 adult horses were included. All the horses underwent to a complete physical examination, blood analysis, and were evaluated for the SIRS status. Blood samples were collected once in healthy horses and at admission time, then 24, 48, 72, and 96 h after admission in sick animals. PCC was evaluated using a method previously described. Data were statistically analyzed to verify differences in PCC between healthy vs. SIRS positive or SIRS negative horses at all sampling time. The receiver operating characteristic (ROC) curve was performed to verify sensitivity and specificity of PCC in the diagnosis of SIRS-positive and SIRS negative horses. The healthy horses were standardbred mares with a median age of 8.5 years. The sick horses were 31/54 females, 16/54 geldings, and 7/54 stallions of different breeds and with a median age of 12 years old. Eight out of 54 sick horses were SIRS negative, while 46/54 were SIRS positive. Statistically significant differences were obtained between healthy and SIRS positive horses, while no differences were observed between healthy and SIRS negative horses at any sampling time. The best cutoff value of PCC to discriminate between SIRS positive, SIRS negative, and healthy horses, the sensitivity and specificity of cutoff point, the area under receiver operating characteristic curve, the 95% confidence intervals, and the likelihood ratio were reported. We found higher PCC values in sick SIRS-positive horses vs. healthy ones with a decrement over time, while no differences at admission, nor during the observational period, were obtained in sick but SIRS-negative horses. The value of 0.049 nmol/ml/mg is reported as a potential cutoff for the diagnosis of SIRS positivity vs. healthy horses with a sensibility of 74.5% and a specificity of 72.2%. In conclusion, PCC seems to be a sensitive and specific marker for SIRS in horses.

Combined effects of salinity changes and salicylic acid exposure in Mytilus galloprovincialis.

Pharmaceuticals and Personal care products (PPCPs) are frequently released into several marine matrices, representing significant environmental and ecotoxicological risks. Among the widest spread PPCPs in aquatic systems is Salicylic acid (SA), with known negative effects on marine and freshwater species. Nevertheless, the toxicity resulting from these emerging pollutants, including SA, together with climate change has still received little attention up to date. Among climate change related factors salinity is one that most affects aquatic organisms. To better understand the combined impacts of SA and salinity, the present study evaluated the biochemical alterations induced in Mytilus galloprovincialis mussels exposed to SA and different salinity levels, acting individually and in combination. The effects observed clearly highlighted that cellular damages were mainly observed at higher salinity (35), with no additive or synergistic effects derived from the combined presence of SA. Higher antioxidant capacity of mussels in the presence of SA may prevent increased LPO levels in comparison to uncontaminated mussels. Nevertheless, in the presence of SA mussels revealed loss of redox balance, regardless of the salinity level. Furthermore, mussels exposed to SA at control salinity showed increased metabolic capacity which decreased when exposed to salinities 25 and 35. These findings may indicate the protective capacity of mussels towards higher stressful conditions, with lower energy reserves expenditure when in the presence of SA and salinities out of their optimal range. Although limited cellular damages were observed, changes on mussel's redox balance, antioxidant mechanisms and metabolism derived from the combined exposure to SA and salinity changes may compromise mussel's growth and reproduction. Overall, the present study highlights the need to investigate the impacts induced by pollutants under present and future climate change scenarios, towards a more realistic environmental risk assessment.

Biochemical and physiological responses of two clam species to Triclosan combined with climate change scenario.

Ocean acidification and warming are among the man-induced factors that most likely impact aquatic wildlife worldwide. Besides effects caused by temperature rise and lowered pH conditions, chemicals of current use can also adversely affect aquatic organisms. Both climate change and emerging pollutants, including toxic impacts in marine invertebrates, have been investigated in recent years. However, less information is available on the combined effects of these physical and chemical stressors that, in nature, occur simultaneously. Thus, this study contrasts the effects caused by the antimicrobial agent and plastic additive, Triclosan (TCS) in the related clams Ruditapes philippinarum (invasive) and Ruditapes decussatus (native) and evaluates if the impacts are influenced by combined temperature and pH modifications. Organisms were acclimated for 30 days at two conditions (control: 17 °C; pH 8.1 and climate change scenario: 21 °C, pH 7.7) in the absence of the drug (experimental period I) followed by a 7 days exposure under the same water physical parameters but either in absence (unexposed) or presence of TCS at 1 µg/L (experimental period II). Biochemical responses covering metabolic, oxidative defences and damage-related biomarkers were contrasted in clams at the end of experimental period II. The overall picture showed a well-marked antioxidant activation and higher TCS bioaccumulation of the drug under the forecasted climate scenario despite a reduction on respiration rate and unaltered metabolism in the exposed clams. Since clams are highly consumed shellfish, the consequences for higher tissue bioaccumulation of anthropogenic chemicals to final consumers should be alerted not only at present conditions but more significantly under predicted climatic conditions for humans but also for other components of the marine trophic chain.

Behavioral Therapy and Fluoxetine Treatment in Aggressive Dogs: A Case Study.

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.

Impacts of salicylic acid in Mytilus galloprovincialis exposed to warming conditions.

While many studies have been conducted on drug-inducing alterations in the aquatic environment, little is known about their interaction with climate change, such as rising temperatures. To increase knowledge on this topic, Mytilus galloprovincialis mussels were exposed to two different temperatures 17 ± 1 °C (control) and 21 ± 1 °C in the absence and presence of salicylic acid (SA) (4 mg/L) for 28 days. Salicylic acid in the water and tissues was measured and its impact reported through biomarker responses including: energy metabolism (electron transport system (ETS) activity, glycogen (GLY), protein (PROT) and lipids (LIP) contents), oxidative stress markers (activity of the enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx)), glutathione balance between the reduced and the oxidized forms (GSH/GSSG), and damage to membrane lipids (lipid peroxidation - LPO). The mussels responded differently if the stresses imposed were single or combined, with greater impacts when both stressors were acting together. Contaminated mussels exposed to high temperatures were unable to increase their metabolic capacity to restore their defence mechanisms, reducing the expenditure of LIP. In the presence of SA and increased temperature antioxidant defences respond differently, with higher SOD levels and inhibition of CAT. The present study highlights not only the negative impact of warming and SA, but especially how temperature increase will promote the impact of SA in M. galloprovincialis, which under predicted climate change scenarios may greatly impair population maintenance and ecosystem biodiversity.

Acute toxicity of ionic liquids for three freshwater organisms: Pseudokirchneriella subcapitata, Daphnia magna and Danio rerio.

The static acute toxicities of 18 ionic liquids (ILs) were determined for three representative freshwater organisms, the cladoceran Daphnia magna, the green alga Pseudokirchneriella subcapitata (formerly known as Selenastrum capricornutum), and the fish Danio rerio (formerly known as zebrafish). The test kit compounds contained three widely used ILs (1-butyl-3-methylimidazolium bis(triflimide), [bmim][Tf(2)N], butylpyridinium bis(triflimide), [bpy][Tf(2)N], and N,N-methylbutylpyrrolidinium bis(triflimide), [bmpyrr][Tf(2)N]) and 15 less common salts. These latter comprised a range of five anions, four positively charged head groups (ammonium, morpholinium, thiophenium, and sulfonium), five 1-methyl-3-alkyl imidazolium derivatives bearing a specific functional group on the longer alkyl chain (Cl, OH, or (CH(3))(3)Si) and three imidazolium derivatives characterized by the presence of a hydrogen atom on the imidazolium nitrogen ("Brønsted acidic imidazolium"-based ILs). Generally, long-chain ammonium salts showed higher toxicity to algae, cladocerans, and fish, whereas very low toxicities characterized sulfonium- and morpholinium-based ILs. In imidazolium-based ILs, the substitution of one or two carbon atoms of the longer alkyl chain with a more electronegative atom (chlorine or oxygen) reduced the acute toxicity for algae and cladocerans. Low toxicity also characterized the "Brønsted acidic imidazolium"-based ILs. Structural information for a rational designer of safer ILs can be obtained from these studies.

The influence of temperature on the effects induced by Triclosan and Diclofenac in mussels.

Little is known about the consequences of exposure to pharmaceuticals and personal care products (PPCPs) in elevated temperatures associated with climate change. To increase the knowledge on this topic, Mytilus galloprovincialis mussels were exposed to 1.0 µg/L of either the antimicrobial Triclosan (TCS) or the anti-inflammatory drug Diclofenac (DIC), at control (17 °C) and 4 °C raised (21 °C) temperatures for 28 days. Triclosan and DIC concentrations in the water and tissues were subsequently measured and related to biomarker responses including: energy metabolism (electron transport system (ETS) activity, glycogen and protein reserves), oxidative stress markers, glutathione balance between the reduced and the oxidised form (GSH/GSSG), and damage to proteins and lipids. Mussels responded to the increase in temperature and drug exposure by lowering their metabolic rate (decreased ETS), increasing their endogenous reserves and antioxidant defences, thus preventing oxidative stress damage, with the exception of DIC exposure at the higher temperature. In all cases, GSH/GSSG ratio was reduced in detriment of the antioxidant form at both PPCPs exposures and elevated temperature with no additive effect due to combined stressors. Overall, either drug exposure or increased temperature could compromise the ability of mussels to withstand further insults.

Evaluation of glomerular filtration rate estimation by means of plasma clearance of iohexol in domestic rabbits (Oryctolagus cuniculus).

OBJECTIVE: To evaluate glomerular filtration rate (GFR) estimation by means of plasma clearance of iohexol (IOX) in domestic rabbits and to assess accuracy of limited-sampling models for GFR estimation. ANIMALS: 6 healthy domestic rabbits (Oryctolagus cuniculus). PROCEDURES: Each rabbit received IOX (64.7 mg/kg [0.1 mL/kg], IV), and blood samples were collected at predetermined times before and after administration. Plasma IOX concentration was determined by high-performance liquid chromatography. The pharmacokinetics of IOX was determined by a noncompartmental method. For each rabbit, plasma clearance of IOX was determined by dividing the total IOX dose administered by the area under the concentration-time curve indexed to the subject's body weight. The GFR estimated from the plasma IOX concentration at 6 sampling times (referent model) was compared with that estimated from the plasma IOX concentration at 5 (model A), 4 (model B), and 3 (models C, D, and E) sampling times (limited-sampling models). RESULTS: Mean ± SD GFR was 4.41 ± 1.10 mL/min/kg for the referent model and did not differ significantly from the GFR estimated by any of the limited-sampling models. The GFR bias magnitude relative to the referent model was smallest for model D in which GFR was estimated from plasma IOX concentrations at 5, 15, and 90 minutes after IOX administration. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that plasma clearance of IOX was a safe, reliable, accurate, and clinically feasible method to estimate GFR in domestic rabbits. Further research is necessary to refine the method.