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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.

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 (EC

With the increase of UV filters usage and consequent release into aquatic environments, the concerns about their potential ecological risks are also increasing. According to this, in the present study, adult polychaetes of the species Ficopomatus enigmaticus were chronically exposed to three concentrations (0.01, 0.1 and 0.5 mg/L) of organic and inorganic filters (Ethylhexyl methoxycinnamate (EHMC) and nanoparticulate Zinc oxide (nZnO), respectively) in order to analyse biochemical responses related to cellular damage, antioxidant defence, biotransformation mechanisms and, lastly, neurotoxicity. Despite major lipid peroxidation caused by EHMC was observed, both UV filters have produced the same response patterns. In details, a clear concentration-dependent activation of glutathione S-transferases and a significant decrease of acetylcholinesterase levels defined an important neurotoxic effect was observed for both contaminants. These results become important to expand the limited scientific literature on biochemical responses of marine and brackish water invertebrates to organic and inorganic UV filters.

Pharmaceuticals drugs are Contaminants of Emerging Concern (CECs) and are continuously discharged into the environment. As a result of human and veterinary use, these substances are reaching aquatic coastal systems, with limited information regarding the toxic effects of these compounds towards inhabiting organisms. Among CECs are pharmaceuticals like 17 alpha-ethinylestradiol (EE2), which is a synthetic hormone with high estrogenic potency. EE2 has been increasingly found in different aquatic systems but few studies addressed its potential toxicity to marine wildlife, in particular to bivalves. Therefore, the aim of the present study was to evaluate the influence of temperature (17 oC-control and 21 °C) on the potential effects of EE2 on the mussel Mytilus galloprovincialis. For this purpose, mussels were exposed to different concentrations of EE2 (5.0; 25.0; 125.0 and 625 ng/L), resembling low to highly polluted sites. Mussels exposed to each concentration were maintained under two temperatures, 17 and 21 °C, which represent actual and predicted warming conditions, respectively. After 28 days, oxidative stress status, metabolism related parameters, neurotoxicity and histopathological alterations were measured. The results obtained clearly showed an interactive effect of increased temperature and EE2, with limited antioxidant and biotransformation capacity when both stressors were acting together, leading to higher cellular damage. The combination of both stressors also enhanced mussels’ metabolic capacity and neurotoxic effects. Nevertheless, loss of redox balance was confirmed by the strong decrease of the ratio between reduce glutathione (GSH) and oxidized glutathione (GSSG) in contaminated mussels, regardless the temperature. Histopathological indexes in contaminated mussels were significantly different from the control group, indicating impacts in gills and digestive glands of mussels due to EE2, with higher values observed at 21 °C. Overall, this study demonstrates that of EE2 represents a threat to mussels and predicted warming conditions will enhance the impacts, which in a near future might result in impairments at the population and community levels.

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.

Background There is no commonly shared severity score for myxomatous mitral valve disease (MMVD) based on routinely acquired echocardiographic variables. Hypothesis/Objectives To propose an easy‐to‐use echocardiographic classification of severity of MMVD in dogs. Animals Five hundred and sixty dogs with MMVD. Methods This was a retrospective, multicenter, observational study. The proposed Mitral INsufficiency Echocardiographic (MINE) score was based on 4 echocardiographic variables: left atrium‐to‐aorta ratio, left ventricular end‐diastolic diameter normalized for body weight, fractional shortening, and E‐wave transmitral peak velocity. Specific echocardiographic cutoffs were defined based on previous prognostic studies on MMVD, and severity scores were assigned as follows: mild (score: 4‐5), moderate (score: 6‐7), severe (score: 8‐12), late stage (score: 13‐14). Results Median survival time was significantly different (P 8 was predictive of cardiac death (area under the curve = 0.85; P

Background: Although the influence of copper ([Cu]) on chronic hepatitis (CH) has been widely studied in dogs, little information is available about the accumulation of other metals. Hypothesis/Objectives: We assessed the concentration of lead ([Pb]) in the livers of dogs with CH with or without abnormal hepatic [Cu] to establish if any association existed between [Pb] and either hematologic or biochemical variables, fibrosis, necrosis and inflammation of the liver on histology. Animals: Thirty-four dogs with CH that had hepatic [Cu] and [Pb] determined. Methods: Retrospective review of medical records of dogs with CH and hepatic [Cu] and [Pb]. Chronic hepatitis was defined using current American College of Veterinary Internal Medicine consensus statement guidelines. Hepatic [Cu] and [Pb] were determined using square wave anodic stripping voltammetry. Dogs were divided into 2 groups based on [Cu]

The influence of salinity on the effects of sodium lauryl sulfate (SLS) was evaluated using the Mediterranean mussel Mytilus galloprovincialis, exposed for 28 days to SLS (control-0.0 and 4.0 mg/L) under three salinity levels (Control-30, 25 and 35). The effects were monitored using biomarkers related to metabolism and energy reserves, defence mechanisms (antioxidant and biotransformation enzymes) and cellular damage. The results revealed that non-contaminated mussels tended to maintain their metabolic capacity regardless of salinity, without activation of antioxidant defence strategies. On the contrary, although contaminated mussels presented decreased metabolic capacity at salinities 25 and 35, they were able to activate their antioxidant mechanisms, preventing cellular damage. Overall, the present findings indicate that SLS, especially under stressful salinity levels, might potentially jeopardize population survival and reproduction success since reduced metabolism and alterations on mussels’ antioxidant mechanisms will impair their biochemical and, consequently, physiological performance.

Ochratoxin A (OTA) is a mycotoxin produced by Aspergillus and Penicillium, capable of contaminating several foodstuffs. OTA damages primarily the kidneys, and is suspected to be a carcinogenic substance, thus maximum levels for OTA in foodstuffs have been established in the EU. Italian Ministry of Health suggested a maximum level of 1 μg/kg OTA in pork meat and derived products. In this study, OTA concentrations in liver, kidney, and muscle of 64 wild boars (Sus scrofa) killed in two areas (area A and B) of Parma province (northern Italy), characterized by different habitat types, were assessed by HPLC-FLD technique. OTA was detected in 54% liver, 52% kidney, and 16% muscle samples. OTA levels were significantly higher in liver and kidney compared with muscle, and were above 1 μg/kg in 19 liver, 17 kidney, and 4 muscle samples. OTA levels in wild boars from area A resulted significantly higher with respect to those from area B, suggesting an environmental influence on OTA contamination in wild boars. This study seems to confirm that wild boar meat is a potential source of OTA, thus monitoring the presence of this mycotoxin in game meat might be recommended to prevent risks for human health.

pages, 7 figures, 2 tables, 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, Francesca Coppola benefited from a PhD grant (SFRH/BD/118582/2016) given by the National Funds through the Portuguese Science Foundation (FCT), supported by FSE and Programa Operacional Capital Humano (POCH) e European Union. Rosa Freitas was funded by national funds (OE), through FCT – Fundação para a Ciência e a Tecnologia, I.P., in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19. Thanks are due for the financial support to CESAM (UIDB/50017/2020 + UIDP/50017/2020), to FCT/MEC through national funds, and co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020. This work acknowledges the contribution of the project BISPECIAl: BIvalveS under Polluted Environment and ClImate chAnge PTDC/CTA-AMB/28425/2017 (POCI-01-0145-FEDER-028425) funded by FEDER, through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI), and by national funds (OE), through FCT/MCTES; the AimCost project (ref CGL2016-76332-R MINECO/FEDER/UE) and The RED RIESCOS (CYTED) ref. 419RT0578, With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI)

Background: Homocysteine (HCY) was evaluated in healthy and chronic enteropathic dogs, however no studies on dogs with immunosuppressant-responsive enteropathy are available. Objectives: The aim was to evaluate serum HCY concentrations and its prognostic role in dogs with immunosuppressant-responsive enteropathy compared to healthy dogs. Methods: Serum HCY concentration was statistically compared between 24 healthy dogs and 29 dogs with immunosuppressant-responsive enteropathy. Correlation analyses between serum total protein, albumin (ALB), C-reactive protein (CRP), folate and cobalamin, and serum HCY concentration were performed in immunosuppressant-responsive enteropathic dogs. Results: The associations between serum HCY concentration and clinical, histological, endoscopic scores and follow-up were evaluated. Mean serum HCY concentration was higher in immunosuppressant-responsive enteropathic dogs compared to control dogs (30.22 ± 8.67 μmol/L vs. 5.26 ± 2.78 μmol/L; p < 0.0001). No association between serum HCY concentration and total protein, ALB, CRP, folate concentration as well as, clinical score, histological and endoscopic scores was found. A negative correlation between serum HCY concentration and cobalamin was noted (p = 0.0025, r = −0.54). No significant difference in HCY was found between responsive and non-responsive dogs or between survivors and non-survivors. Conclusions: Although, serum HCY concentration was higher in immunosuppressant-responsive enteropathy, its prognostic value remains unclear. However, further prospective, large-scale studies are warranted to better investigate the possible prognostic role of HCY in immunosuppressant-responsive enteropathic dogs.

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. published

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).

11 pages, 6 figures, 1 table, supplementary data https://doi.org/10.1016/j.ecoenv.2019.109899, 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, Francesca Coppola benefited from a PhD grant (SFRH/BD/118582/2016) given by the National Funds through the Portuguese Science Foundation (FCT), supported by FSE and Programa Operacional Capital Humano (POCH) e European Union. Rosa Freitas was funded by national funds (OE), through FCT – Fundação para a Ciência e a Tecnologia, I.P., in the scope of the framework contract foreseen in the numbers 4, 5 and 6 of the article 23, of the Decree-Law 57/2016, of August 29, changed by Law 57/2017, of July 19. Thanks are due for the financial support to CESAM (UID/AMB/50017/2019), to FCT/MEC through national funds, and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020. This work was also financially supported by the project BISPECIAl: BIvalveS under Polluted Environment and ClImate chAnge (POCI-01-0145-FEDER-028425) funded by FEDER, through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI), and by national funds (OE), through FCT/MCTES; and by the Spanish Ministry of Economy, Industry and Competitivity: AimCost project (ref CGL 2016-76332-R MINECO/FEDER/UE), With the funding support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S), of the Spanish Research Agency (AEI)

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.

Mixture of contaminants often determine biological responses of marine species, making difficult the interpretation of toxicological data. The pharmaceutical 17 alpha-ethinylestradiol (EE2) and the surfactant Sodium Lauryl Sulfate (SLS) commonly co-occur in the marine environment. This study evaluated the effects of EE2 (125.0 ng/L) and SLS (4 mg/L), acting individually and combined, in the mussel Mytilus galloprovincialis. Contaminated mussels closed their valves for longer periods than control ones, especially in the presence of both contaminants, with longer closure periods immediately after spiking compared to values obtained one day after spiking. Nevertheless, males and females increased their metabolism when in the presence of both contaminants (males) and SLS (females), and independently on the treatment males and females were able to activate their antioxidant and biotransformation defences. Although enhancing defences mussels still presented cellular damage and loss of redox balance, especially noticed in the presence of EE2 for males and SLS for females. Histopathological damage was found at mussel's gills in single and mixture exposure, and qPCR analysis revealed a clear estrogen receptor expression with no additive effect due to combined stressors. The results obtained highlight the harmful capacity of both contaminants but further research on this matter is needed, namely considering different climate change scenarios.

The presence of pharmaceuticals in the aquatic environment is an ongoing concern. However, the information regarding their effects under different climate change scenarios is still scarce. 17α-ethinylestradiol (EE2) is widely present in different aquatic systems showing negative impacts on aquatic organisms even when present at trace concentrations (≈1 ng/L). Nevertheless, its impact on bivalves is poorly understood, especially considering the influence of climate change factors. This study aimed to assess the toxicological impacts of EE2 under current and predicted warming scenarios, in the edible clam Ruditapes philippinarum. For this, clams were exposed for 28 days to different EE2 concentrations (5, 25, 125, 625 ng/L), under two temperatures (17 °C (control) and 21 °C). Drug concentrations, bioconcentration factors and biochemical parameters, related to oxidative stress and energy metabolism, were evaluated. Results showed that under actual and predicted temperature scenarios EE2 concentrations led to a disturbance in redox homeostasis of the clams, characterized by an increase in oxidized glutathione in contaminated organisms compared to control ones. Nevertheless, clams were capable to cope with the stressful conditions, activating their defence mechanisms (especially at the highest exposure concentration and in particular at increased temperature), and no oxidative damage occured. Although limited effects were observed, the present findings indicate that under both temperatures contaminated clams altered their biochemical performance, which can impair their sensitivity and protection capacity to respond to other environmental changes and/or affect their capacity to grow and reproduce. The results presented here highlight the need for further research on this thematic, considering that climate change is an ongoing problem, and the levels of some pharmaceutical drugs will continue to increase in marine/estuarine environments.

Procalcitonin (PCT) has emerged as a promising biomarker for the rapid identification of sepsis both in human and veterinary medicine. Nevertheless, the only analytical method currently available for the detection of PCT in veterinary species, is represented by immunoassays, useful only for research purposes. In this work, we report the development of two biosensors which utilize molecularly imprinted polymers (MIPs) for the detection of canine and equine PCT. Dopamine (DA) and norepinephrine (NE) were used as monomers for the synthesis of the MIP films on surface plasmon resonance (SPR) gold chips and the imprinting efficiency of canine and equine PCT in terms of binding affinity toward the analyte, selectivity, and sensitivity were compared. After optimization in buffer conditions, PCTs calibration was successfully achieved also in animal plasma, with good specificity and reproducibility. More effective protein binding and imprinting was obtained with polynorepinephrine (PNE) for both PCTs, and the SPR biosensors were able to detect the biomarkers in plasma with a LOD of 15 ng mL−1 and 30 ng mL−1 respectively for equine and canine PCT.

The aim of the present study was to investigate retrospectively the plasma concentration of alpha-tocopherol in dogs with naturally acquired chronic kidney disease (CKD), at different stages of severity. Forty dogs (CKD group) with different stages of CKD (IRIS 1 n=12, IRIS 2 n=8, IRIS 3 n=11, IRIS 4 n=9) and 20 clinically healthy dogs were considered. Plasma alpha-tocopherol was assessed in both groups through high performance liquid chromatography (HPLC). Dogs of CKD group showed significantly lower (p=0.0002) levels of plasma alpha-tocopherol compared with clinically healthy dogs. A significant difference (p0.04) in the number of patients with plasma alpha-tocopherolor ≤ 21.5 ppm was found in CKD patients at different stages of severity. No significant correlation between plasma levels of alpha-tocopherol and plasma creatinine was found. In the present study, dogs affected by spontaneous CKD showed significantly lower plasma concentrations of alpha-tocopherol compared with clinically healthy dogs. Plasma alpha-tocopherol deficiency seems to be more severe in IRIS stage 1 and 4, compared with IRIS stage 2 and 3. The finding of marked alpha-tocopherol deficiency in patients in IRIS stage 1 should encourage further studies on the early use of prescription renal diet and antioxidant in this group of patients.

Background Asymmetric dimethylarginine (ADMA) has been proposed as a severity marker in humans with acute pancreatitis (AP). Objectives To evaluate ADMA in dogs with AP compared with healthy dogs and its association with severity of disease, mortality, and indicators of the systemic inflammatory response syndrome (SIRS), including serum C‐reactive protein (CRP) concentration, WBC count, and band neutrophils. Animals Fifty‐four dogs with AP and a control group (CG) of 28 healthy dogs. Methods Cohort study including dogs with AP diagnosed using clinical and laboratory variables, abnormal canine pancreatic lipase (cPL) concentration, and compatible abdominal ultrasound examination findings performed within 48 hours of admission. Canine AP severity (CAPS) was calculated. Serum concentration of ADMA was measured using high performance liquid chromatography. Blood donor‐, staff‐, and student‐owned dogs were enrolled in the CG. Results Dogs with AP had higher median admission serum ADMA concentrations compared with the CG (62 versus 48.5 μg/dL; P = .003). Dogs with CAPS ≥11 had higher serum ADMA concentrations than did dogs with CAPS

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.6 healthy domestic rabbits (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).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.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.

BACKGROUND There is good diagnostic accuracy of smartphone-based ECG in the evaluation of heart rate (HR), heart rhythm, and ECG values for humans, horses, dogs, and cats. OBJECTIVES Assess feasibility and reliability of a smartphone ECG device to evaluate HR expressed as beats per minute and ECG values in cows. ANIMALS Fifty-five healthy Holstein Friesian cows. METHODS Prospective observational study. A standard base-apex ECG was acquired for 60 seconds in each cow. A smartphone ECG tracing was recorded simultaneously using a single-lead bipolar ECG recorder. All ECGs were reviewed by 1 blinded investigator. The following ECG variables were assessed: HR, P wave duration, PR interval, QRS complex and QT interval (milliseconds), P wave, and QRS complex polarity. Agreement between standard and smartphone ECG was evaluated by Cohen's k test and the Bland-Altman test. RESULTS Smartphone ECG tracings were interpretable in 89% of the recordings. Minimal differences of no clinical relevance were found between standard and smartphone ECG tracings regarding HR and duration of ECG waves and interval. Good agreement found in the evaluation of QRS complex polarity (85%) but not for P wave polarity (k = 0.006). Artifacts were rare but of significantly higher frequency in smartphone ECG compared to standard ECG tracings (22% versus 0%; P = .005). CONCLUSIONS AND CLINICAL IMPORTANCE Good quality single-lead ECG tracings can be recorded by smartphone device in healthy cows. Smartphone tracings are reliable for evaluation of HR and selective ECG variables. Smartphone ECG can represent an additional tool for ECG screening in cows.

10 pages, 5 figures, 2 tables, In the present study Mytilus galloprovincialis mussels were exposed for 28 days to three salinities: 30 (control), 25 and 35. Simultaneously, organisms at each salinity were exposed to either the antimicrobial agent Triclosan (TCS)or the pharmaceutical drug Diclofenac (DIC)at 1 μg/L. Salinity alone and exposure to PPCPs changed mussel's metabolic capacity and oxidative status, but no additive or synergetic effects resulting from the combined exposures were observed. Overall, the metabolic capacity of mussels was decreased when exposed to TCS and DIC under control salinity, which was less pronounced at salinities out of the control level. TCS had a notorious effect over glutathione peroxidase activity while DIC exposure enhanced catalase response. Such defence mechanisms were able to prevent cellular damage but still a clear reduction in GSH/GSSG ratio after PPCPs exposures indicates oxidative stress which could compromise bivalve's performance to further stressing events, Francesca Coppola benefited from PhD grant (SFRH/BD/118582/2016) given by the National Funds through the Portuguese Science Foundation (FCT), supported by FSE and Programa Operacional Capital Humano (POCH) e European Union. Rosa Freitas benefited from a Research position funded by Integrated Programme of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01-0145-FEDER-000018), co-funded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund. Thanks are due for the financial support to CESAM (UID/AMB/50017/2019), to FCT/MEC through national funds, and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020. This work was also financially supported by the project BISPECIAl: BIvalveS under Polluted Environment and ClImate chAnge (POCI-01-0145-FEDER-028425) funded by FEDER, through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI), and by national funds (OE), through FCT/MCTES; and by the Spanish Ministry of Economy, Industry and Competitivity: AimCost project (ref CGL2016-76332-R MINECO/FEDER/UE). Rosa Freitas and Montserrat Solé are also funded by CYTED, through the RED Iberoamericana RIESCOS (419RT0578)

A vast variety of substances currently reaches the aquatic environment, including newly developed chemicals and products. Lack of appropriate analytical methods for trace determinations in aquatic ecosystem compartments and lack of information regarding their toxicity explains existing regulation gaps. However, suspicion of their toxicity assigned them as Contaminants of Emerging Concern (CECs). Among CECs are Pharmaceuticals including Salicylic Acid (SA), which is the active metabolite of acetylsalicylic acid (ASA; aspirin). The aim of the present study was to evaluate the potential effects of SA on the mussel Mytilus galloprovincialis. For this, organisms were exposed for 28 days to different concentrations of SA (0.005; 0.05; 0.5 and 5 mg/L), resembling low to highly polluted sites, after which different physiological and biochemical parameters were evaluated to assess organism’s respiration rate, neurotoxic, metabolic and oxidative stress status. Our results clearly showed that SA strongly reduced the respiration capacity of mussels. Also, SA inhibited the activity of superoxide dismutase (SOD) and catalase (CAT) enzymes, but increased the activity of glutathione peroxidase (GPx) and glutathione-S-transferases (GSTs), which prevented the occurrence of lipid peroxidation (LPO). Nevertheless, oxidative stress was confirmed by the strong decrease of the ratio between reduce glutathione (GSH) and oxidized (GSSG) glutathione in contaminated mussels. Moreover, neurotoxicity was observed in mussels exposed to SA. Overall, this study demonstrates the metabolic, neurotoxic and oxidative stress impacts of SA in M. galloprovincialis, which may result in negative consequences at the population level.

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.

8 pages, 4 figures, 1 table, 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, PhD grant (SFRH/BD/118582/2016) given by the National Funds through the Portuguese Science Foundation (FCT), supported by FSE and Programa Operacional Capital Humano (POCH) e European Union. Rosa Freitas benefited from a Research position funded by Integrated Programme of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro-01-0145-FEDER-000018), co-funded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund. Thanks are also due, for the financial support to CESAM (UID/AMB/50017), to FCT/MEC through national funds, and the co-funding by the FEDER, within the PT2020 Partnership Agreement and Compete 2020; Programa Operacional Competitividade e Internacionalização FEDER (POCI-01-0145-FEDER-028425) BISPECIAl - BIvalves under Polluted Environment and ClImate change; and by the Ministry of Economy, Industry and Competitiveness: AimCost project (ref CGL2016-76332-R MINECO/FEDER/UE)

Pharmaceuticals such as racemate ketoprofen (RS-KP) and its enantiomer, dexketoprofen (S(+)-KP) are highly detectable non-steroidal anti-inflammatory drugs (NSAIDs) in the aquatic environment and therefore are designated as one of the most emerging groups of pollutants that can affect environmental and human health. The potential impact of these pharmaceuticals was assessed for the first time in vitro using a rat hepatocellular carcinoma cell line (H4IIE). Cells were exposed to low and high concentrations of these drugs. Cytotoxicity was determined by MTT reduction assay; CYP1A1 transcriptional and enzymatic levels together with canonical oxidative stress responsive markers (GPx, GR, GST and CAT) were also investigated. Cells exposed to RS-KP and S(+)-KP did not show cytotoxicity effect at the concentrations tested. However, this study highlighted differences between RS-KP and S(+)-KP in most of the evaluated markers, showing compound-, concentration- and time-specific effect patterns which suggest a potential stereo-selective toxicity of these drugs.

The aim of the present study was to evaluate the sensitivity and specificity of urine KIM-1 and urine GGT for the detection of naturally-occurring AKI, compared to healthy control dogs, dogs with stable chronic kidney disease (CKD), and dogs with lower urinary tract disorders (LUTD). The study included AKI grade 1 (n = 21), AKI grade 2 to 5 (n = 11), stable CKD (n = 11), LUTD (n = 15), and healthy dogs (n = 37). Urine KIM-1 (ng/mg) and GGT (U/l) were normalized to urine creatinine (uCr). Statistically significant difference in KIM/uCr (p = 0.0007) and GGT/uCr (p

The diagnosis of sepsis in calves is challenging. Blood culture and clinical signs combined with a complete blood count have been used for the diagnosis of sepsis. Recent literature in humans and animal species has been focused on sepsis-specific biomarkers, such as procalcitonin (PCT), that may more accurately and efficiently diagnose sepsis. The aim of this study was to evaluate plasma PCT concentrations in healthy and septic calves. Twenty healthy control calves and 58 sick calves with septic systemic inflammatory response syndrome (SIRS) based on SIRS score and clinical findings were included. Calves with septic SIRS were further divided in septic SIRS survivors (SSS) and non-survivors (SSNS). Plasma PCT concentrations were measured with a commercial ELISA assay for cattle. A receiver operating characteristic curve was used to determine cut-off values and corresponding sensitivity and specificity for the diagnosis of sepsis. Differences in plasma PCT concentration between groups (control vs. SSS vs. SSNS) were evaluated. Plasma PCT concentrations in healthy calves and those with septic SIRS were 33.3pg/mL (0-44.3pg/mL) and 166.5pg/mL (85.9-233.0pg/mL), respectively (P0.05). These results confirmed an increase in plasma PCT concentrations in calves with septic SIRS, as previously reported in humans and other species.

To test an equine-derived polyvalent viperid antivenom (EPVA) in the treatment of dogs with evidence of viper envenomation.Prospective, multicenter observational study.Veterinary emergency and critical care hospitals.A total of 82 client-owned dogs with progressive clinical signs after viperid snakebite were enrolled in the study.Equine-derived polyvalent viperid antivenom was administered at a dosage of 1 mL/kg body weight, either by IV infusion or SC injection.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%).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.

Ochratoxin A (OTA) is a secondary toxic metabolite synthesized by Aspergillus or Penicillium species, which can contaminate various crops. The International Agency for Research on Cancer (IARC) classified OTA as a group 2B possible human carcinogen. The aim of the present study was to assess OTA concentrations in tissues of wild boar (Sus scrofa L.) from Tuscany (Italy). Over a period of 2 years, samples of muscle, liver, and kidney from 48 wild boars were collected and concentrations of OTA were determined by enzymatic digestion (ED) coupled to high-performance liquid chromatography with a fluorescence detector (HPLC-FLD). The highest concentrations of OTA were found in the kidneys of the 48 wild boars analyzed. No difference in concentrations was found based on years of collection and sex while a significantly higher OTA concentration was found in the kidney of the young wild boars with respect to the adult one. Monitoring the quality of meat destined for transformation is a priority in order to decrease the possibility of toxin carry-over to humans. The present study showed that contamination of wild boar meat products by OTA represents a potential emerging source of OTA.

The increased use of nonsteroidal anti-inflammatory drugs (NSAIDs) has resulted in their ubiquitous presence in the environment. The toxicological properties of these 2 widely prescribed NSAIDs, namely racemic ketoprofen and its enantiomer S(+)-ketoprofen (dexketoprofen), were evaluated, firstly, by acute and chronic toxicity tests using 3 representative model organisms (Vibrio fischeri, Pseudokirchneriella subcapitata, and Ceriodaphnia dubia) and, secondly, by evaluating the responses of biotransformation systems and multidrug resistance-associated proteins (MRP1/MRP2) using the Poeciliopsis lucida hepatocellular carcinoma 1 (PLHC-1) fish hepatic cell line. Toxicity data from both acute and chronic dexketoprofen exposure indicated higher sensitivity through inhibition of bioluminescence and algal growth and through increased mortality/immobilization compared to racemic ketoprofen exposure. The growth inhibition test showed that racemic ketoprofen and dexketoprofen exhibited different effect concentration values (240.2 and 65.6 μg/L, respectively). Furthermore, racemic ketoprofen and dexketoprofen did not exert cytotoxic effects in PLHC-1 cells and produced compound-, time-, and concentration-specific differential effects on cytochrome P450 1A (CYP1A) and glutathione S-transferase levels. For CYP1A, the effects of racemic ketoprofen and dexketoprofen differed at the transcriptional and catalytic levels. Exposure to racemic ketoprofen and dexketoprofen modulated MRP1 and MRP2 mRNA levels, and these effects were also dependent on compound, exposure time, and concentration of the individual drug. The present study revealed for the first time the interactions between these NSAIDs and key detoxification systems and different sensitivity to the racemic mixture compared to its enantiomer. Environ Toxicol Chem 2018;37:201-212. © 2017 SETAC.

Ochratoxin A (OTA) is a mycotoxin produced by secondary metabolism of several fungi belonging to the genera Aspergillus and Penicillium. OTA is potentially nephrotoxic, neurotoxic, immunotoxic and carcinogenic in several animal species and in humans. This toxin has been detected in several human food and animal feed. The aim of this study was to determine OTA in blood samples of healthy and affected by chronic kidney disease (CKD) dogs. CKD group showed higher incidence of OTA-positivity than healthy dogs (96 vs. 56%) and a significantly higher median value of OTA plasma concentration (0.008 vs. 0.144 ng/ml). No significant correlation was observed between OTA levels and creatinine values in CKD dogs. This is the first study regarding OTA detection in plasma samples of healthy and CKD dogs; the presence of this toxin is higher in nephropatic patients but is not yet clear, if it is correlated with progression of the disease.

Objective—To investigate serum calcium-phosphorus concentration product (sCaPP) as a predictor of mortality rate in dogs with chronic kidney disease (CKD). Design—Retrospective case-control study. Animals—31 dogs with definitive CKD and 35 apparently healthy dogs. Procedures—All dogs had been referred for nephrological consultation between December 2008 and December 2010. Dogs with CKD had stable disease for ≥ 3 months. On the basis of glomerular filtration rate < 60 mL/min/m2, 13 of the 35 apparently healthy dogs were subsequently classified as having early CKD. Disease stage among dogs was determined on the basis of plasma creatinine concentration as follows: stage 1, < 123.7 μmol/L (n = 13), stage 2, 123.7 to 176.8 μmol/L (7); stage 3, 185.6 to 442 μmol/L (13); or stage 4, > 442 μmol/L (11). For each dog, serum concentrations of ionized and total calcium and phosphorus were evaluated once; the latter 2 variables were used to determine sCaPP. Results—The sCaPP differed significantly between the 22 healthy dogs and dogs with stage 3 or stage 4 CKD. The proportion of dogs with sCaPP > 70 mg2/dL2 increased with stage of disease. Mortality rate among the 24 dogs with sCaPP > 70 mg2/dL2 was higher than that among the 42 dogs with sCaPP ≤ 70 mg2/dL2. Dogs with sCaPP > 70 mg2/dL2 had a comparatively lower survival rate, and risk of death was 4.2 times as high as risk for dogs with sCaPP ≤ 70 mg2/dL2. Conclusions and Clinical Relevance—For dogs with CKD, sCaPP > 70 mg2/dL2 appeared to be a negative prognostic indicator, which was not influenced by the concomitant serum concentrations of phosphorus and total or ionized calcium.

Escherichia coli are a common inhabitant of the gastrointestinal tract of mammals and birds; nevertheless, they may be associated with a variety of severe and invasive infections. Whereas fluoroquinolones (FQ) have been banned in the United States for use in poultry production, the use of these antimicrobials in poultry husbandry is still possible in the European Union, although with some restrictions. The aim of this study was to investigate the FQ resistance of 235 E. coli isolates recovered from chickens and turkeys. Minimum inhibitory concentrations were determined by a microdilution method, whereas mutations in the quinolone resistance-determining regions of the target genes, gyrA and parC, were detected by a PCR-based method. High resistance rates (>60%) were observed for nalidixic acid, flumequine, and difloxacin, whereas resistance to ciprofloxacin, danofloxacin, enrofloxacin, marbofloxacin, and sarafloxacin was less frequently reported (

Background The diseases most frequent associated with SIRS in adult horses are those involving the gastrointestinal tract. An early diagnosis should be the goal in the management of horses with SIRS. Objective The objective of this study was to evaluate the plasma procalcitonin (PCT) concentration in healthy and SIRS horses to assess differences between the two groups. Animals Seventy-eight horses (30 healthy and 48 SIRS). Methods Prospective in vivo multicentric study. Horses were classified as SIRS if at least 2 of the following criteria were met: abnormal leukocyte count or distribution, hyperthermia or hypothermia, tachycardia, tachypnea. Healthy horses showed no clinical or laboratory signs of SIRS. Plasma PCT concentrations were measured with a commercial ELISA assay for equine species. Results were expressed as mean±standard deviation. T-test for unpaired data was performed between healthy and SIRS group. SIRS group was divided in 4 subgroups and t-test was performed between healthy versus each subgroup. Results PCT concentrations in healthy and SIRS horses were 18.28 ± 20.32 and 197.0 ± 117.0 pg/mL, respectively. T-test showed statistical differences between healthy versus SIRS group and between healthy versus all subgroups. Conclusions and Clinical Importance Results showed an increase in PCT concentration in SIRS horses as previously reported in humans and dogs. PCT could be used as a single assay in equine practice for detection of SIRS.

Nonselective antimuscarinic drugs are clinically useful in several pathologic conditions of horses, but, blocking all muscarinic receptor (MR) subtypes, may cause several side effects. The availability of selective antimuscarinic drugs could improve therapeutic efficacy and safety. We aimed to enlighten the role of different MR subtypes by evaluating the effects of nonselective, and selective M1, M2 and M3 MR antagonists on the contractions of horse jejunum. Segments of circular muscle of equine jejunum, were put into organ baths, connected to isotonic transducers, and the effects on ACh concentration-response curves, and on electrical field stimulation (EFS)-evoked contractions of intestinal preparations, induced by nonselective or selective MR antagonists, compared to pre-drug level, were studied. Atropine (nonselective MR antagonist), pirenzepine (selective M1 antagonist), and p-FHHSiD (selective M3 antagonist) competitively antagonized ACh (pA2=9.78±0.21; 7.14±0.25 and 7.56±0.17, respectively). Methoctramine (selective M2 antagonist) antagonized ACh in a concentration-unrelated fashion; however, it competitively antagonized carbachol, a nonselective muscarinic agonist (pA2=6.42±0.23). Atropine dose-dependently reduced EFS-evoked contractions, reaching a maximal effect of -45.64±6.54%; the simultaneous block of neurokinin receptors, almost completely abolished the atropine-insensitive contractions. p-FHHSiD dose-dependently reduced EFS-induced contractions, while pirenzepine caused a minor decrease. Methoctramine, ineffective up to 10-7M, enhanced the contractions at 10-6M; the block of neurokinin receptors abolished the increase of contraction. Cholinergic contractions of horse jejunum are mainly mediated by M3 receptors; M2 selective antagonists seem to scarcely affect cholinergic, and to enhance neurokininergic contractions of equine jejunum, thus their use entails a lower risk of causing intestinal hypomotility, compared to nonselective drugs.

Background A clinical trial was conducted in order to assess the efficacy of rifaximin, a broad-spectrum antibiotic with negligible gastrointestinal absorption, in comparison with metronidazole, a commonly employed antimicrobial drug, in dogs with chronic enteropathy. Twenty-four pet dogs were randomly enrolled into two different groups: MET group (10 dogs) and RIF group (14 dogs). Dogs of MET group received metronidazole 15 mg/kg q12h for 21 days by oral route, whereas dogs of RIF group, were given rifaximin 25 mg/kg q12h for 21 days by oral route. Clinical signs of disease were evaluated the day before the beginning of drug administration (D0), and at the end of treatment (D21), by means of Canine IBD Activity Index (CIBDAI). Blood levels of C-reactive protein (CRP) at D0 and D21 were also measured, as another parameter of treatment efficacy. The primary outcome measure of efficacy was the complete remission at D21, defined as a 75 % or greater decrease of CIBDAI; secondary outcome measures were the variation of mean CIBDAI scores, of mean CRP serum levels, and any observed adverse effect from D0 to D21. Results Treatment with metronidazole or rifaximin greatly improved the clinical signs of disease in each group: in MET group the complete remission was achieved in 8 of 10 dogs (80.0 %), and partial remission in 2 subjects (20.0 %). In RIF group, 12 of 14 dogs showed complete remission (85.7 %), and the remaining 2 dogs were in partial remission (14.3 %). There were also significant decreases of CIBDAI scores (P = 0.002 and P = 0.0002 for MET and RIF, respectively), and CRP levels (P = 0.002 and P = 0.0001 for MET and RIF, respectively) compared to pre-treatment values in both groups. No significant difference, however, was found when comparing MET and RIF groups. No relevant side-effect was reported during the trial with either drugs. Conclusions The present study showed, for the first time, that oral rifaximin could represent an effective alternative to metronidazole for the induction of clinical remission in dogs with chronic enteropathy. Electronic supplementary material The online version of this article (doi:10.1186/s12917-016-0851-0) contains supplementary material, which is available to authorized users.

Endotoxemia represents a significant clinical and economic problem for the equine industry. This study assesses the kinetics of soluble CD14 (sCD14), chemokine (CC motif) ligand 2 (CCL2), interleukin 10 (IL-10) and plasma procalcitonin (PCT) in healthy horses after the intravenous infusion of lipopolysaccharide (LPS). The aim was to contribute to the basic understanding of the equine species-specific kinetics of these molecules in response to LPS exposure, which could support further findings in clinical studies and identify valuable inflammatory biomarkers for equine practice. Eleven healthy horses were involved in this experimental in vivo study. Horses were classified as healthy before the LPS infusion. After the pre-infusion blood collection (T0), all horses received an infusion of E. coli endotoxin (30ng/kg over 30min). Data and samples were collected 1h (T1), 2 (T2), 3 (T3) and 24h (T24) after infusion. Plasma sCD14, CCL2 and IL-10 were evaluated with a fluorescent bead-based assay, while PCT was evaluated with an equine PCT ELISA assay. A one-way ANOVA test was performed between each blood-sampling time for PCT, sCD14 and IL-10, and a Friedman test was performed for CCL2. Plasma PCT, IL-10 and CCL2 concentrations increased statistically significantly at T1, T2 and T3 compared to T0. No statistically significant differences were found between plasma IL-10 and CCL2 concentrations between T0 vs T24, although plasma PCT values remained high 24h after LPS infusion. Plasma sCD14 concentration showed no statistically significant differences for any of sampling times. Our results demonstrate that LPS injection into healthy horses results in PCT, CCL2 and IL-10 increases in plasma without an increase in sCD14. The increases in PCT, CCL2 and IL-10 are related to the inflammatory response induced by circulating lipopolysaccharide.

Low intensity exercise increases insulin-stimulated glucose uptake in skeletal muscle and decreases its plasma concentration. In this study, plasma insulin and glucose concentrations were evaluated 5min before and 5, 15, 25, 35, 45 and 60min after an IV bolus of glucose in 12 Thoroughbreds before and after 1 month of submaximal aquatraining exercise, monitored using heart rate and blood lactate. Plasma glucose concentrations were evaluated using a colorimetric enzymatic method, and plasma insulin concentrations with a solid-phase radioimmunoassay method. Pre-training plasma glucose concentrations at 15, 25 and 35min, area under the glucose curve and peak glucose concentration were significantly higher than post-training values (P

Disposable electrochemical sensor for rapid measurement of heavy metals in fish by square wave anodic stripping voltammetry (SWASV) V. Meucci & L. Intorre & C. Pretti & S. Laschi & M. Minunni & M. Mascini Published online: 9 July 2009 # Springer Science + Business Media B.V. 2009

Tetracyclines, which represent one of the most commonly used antibiotics for poultry, are known to be deposited in bones, where they can remain, despite the observation of appropriate withdrawal times. The aim of the study was to determine the concentration of oxytretracycline (OTC) residues in the bone and muscle of chickens, following the oral administration of a commercially available liquid formulation, and to test their cytotoxic effects on an in vitro cell culture model. Seventy-two 1-day-old broiler chickens were randomly allotted into 2 groups (control and treated animals). OTC (40 mg/kg BW) was administered via drinking water during the 1 to 5 and 20 to 25 days of life periods. At the end of the trial, the birds were slaughtered and the OTC residues in the target tissues were measured by means of liquid chromatography (LC) - tandem mass spectrometry (MS/MS). Cytotoxicity was assessed by evaluating the pro-apoptotic effect of the bone residues on the K562 erythroleukemic line and on the peripheral blood mononuclear cells (PBMC). In all the animals, the OTC residues in the muscle were far below the established MRL of 100 μg/kg. The OTC levels in the bones of the treated animals were instead found in the parts per million (ppm) range. Cell cytotoxicity was assessed by evaluating the pro-apoptotic effect of OTC bone residues on the haematopoietic cell system. This in vitro system has revealed a significant pro-apoptotic effect on both the K562 cell line and PBMC cultures. This result suggests potential human and animal health risks due to the entry of tetracycline residues contained in the bones of treated livestock into the food-chain. This could be of concern, particularly for canine and feline diets, as meat, bone meal, and poultry by-products represent some of the main ingredients of pet foods, especially in the case of dry pet food. Further studies are needed to define the underlying mechanisms of cytotoxicity and to evaluate the in vivo toxicological implications due to the observed in vitro effects.

Induction of blood plasma vitellogenin (Vtg) and zona radiata proteins (Zr-proteins) in male and juvenile of oviparous vertebrates was proposed and shown to be sensitive biomarkers for exposure to estrogen mimic. The time- and dose-dependent expression of Vtg and Zr-proteins in nonylphenol (NP) exposed juvenile Atlantic salmon (Salmo salar) is reported. Fish were exposed continuously to waterborne nonylphenol at 5, 15 and 50 μg/L. Blood and surface mucus samples were collected after 3 and 7 days post-exposure. Nonylphenol-induced plasma and surface mucus levels of Vtg and Zr-protein were analysed using immunochemical methods (Western blot and enzyme-linked immunosorbent assay; ELISA). Both Vtg and Zr-protein levels in plasma and surface mucus showed similar and parallel nonylphenol-induced expression patterns after waterborne nonylphenol exposure and in a concentration- and time-dependent manner. Zr-proteins were significantly induced at the lowest concentration of nonylphenol after 3 and 7 days of exposure both in plasma and in surface mucus. We conclude that the detection of Vtg and Zr-proteins directly in the surface mucus of fish, and the correlation of these values with plasma protein biomarker values in xenoestrogen-treated fish represents a sensitive non-invasive system for the detection of these known endocrine disruptor biomarkers. The demonstration of detectable Vtg and Zr-protein levels from surface mucus is a potential biomarker for estrogenic compounds, and their presence should be considered as an improvement in the methods for detection of endocrine disrupting chemicals (EDCs) and related pollutants in the environment.

A high-performance liquid chromatography method to determine iohexol (IOX) and p-aminohippuric acid (PAH) in the plasma of dogs is evaluated according to recovery, reproducibility, and linearity utilizing a gradient pump. The mobile phase consists of 50mM sodium dihydrogen phosphate with 0.5mM tetrabutylammonium chloride, the pH is adjusted to 4.1, methanol is added to the final ratio of 90:10 (v/v), the flow rate is set at 1 mL/min, and separation is achieved with an ODS2 Luna column. The UV detector is set at 254 nm. IOX and PAH are used for evaluation of the effective renal plasma flow (ERPF) and glomerular filtration rate (GFR). The present method tested in three dogs demonstrates the accuracy in the evaluation of ERPF and GFR. Because of its precision and simplicity and low cost, it can be considered a good tool for ERPF and GFR in small animal practice.

The safety of azamethiphos (AZA), an organophosphorous insecticide and the active ingredient of Salmosan, was evaluated in the European eel, seabass and rainbow trout. Fish were bathed in 0.1 ppm AZA for a period of 60, 120 or 240 min. After termination of each treatment fish were transferred to clean aquaria and randomly sampled over 21 days. Compared to controls, brain acetylcholinesterase (AChE) was inhibited up to 44, 56 and 62% in eels, seabass and trout, respectively, with the inhibition being significant for up to 4 days in eels and seabass and 7 days in trout. As result of the AChE depression, fish displayed motor hyperactivity and erratic jumping at the onset of treatment. Mortality was observed only in trout following exposure for 240 min. A variable correlation observed among species between the level of exposure, the reduced activity of brain AChE and the signs of toxicity suggest that brain AChE should be considered as an indicator of exposure rather than as an index of toxicity of AZA. The present data indicate that at the therapeutic dosage of 0.1 ppm AZA for 1h can be safely used in eels, seabass and trout. The extended treatment times up to 240 min were equally safe for eels and seabass but not for trout.

Background: Detection of nonsteroidal anti-inflammatory drugs (NSAIDs) in equine plasma is a significant analytical problem in veterinary anti-doping controls. Results: A new HPLC method coupled to selective extraction with molecularly imprinted polymers was developed for the simultaneous determination in equine plasma of the NSAIDs phenylbutazone, flunixin, oxyphenbutazone, ketoprofen and naproxen. The analytical performances of the method have been evaluated both in standard solutions and equine plasma samples. Recovery: Molecularly imprinted polymers solid-phase extraction for all NSAIDs was >94% with intra-day values below 15.0% and inter-day values below 20%. Method quantification limit was 0.01 μg/ml. Conclusion: The developed method could be considered as a useful alternative to existing screening methods for the determination of NSAIDs in the context of studies of pharmacokinetics and anti-doping controls.

The efficacy of enrofloxacin (ENRO) was evaluated against multidrug-resistant avian pathogenic Escherichia coli correlating the minimum inhibitory concentrations (MIC) of 235 E. coli field strains with its pharmacokinetics (PK) in 50 healthy turkeys (5 groups) with a PK/pharmacodynamic approach. The treatments were as follows: a) single oral gavage and b) single subcutaneous (SC) treatment at the recommended dose of 10 mg/kg; c) single oral gavage, d) 5 d of 10-h pulsed water medication, and e) 5 d of 24-h continuous water medication at the doubled dose of 20 mg/kg. Blood samples were collected at established times over 24 h. Plasma was analyzed using a liquid chromatography tandem mass spectrometry method that was validated in house. A monocompartmental and a noncompartmental model were applied to the data to obtain the PK results. After gavage administration, the mean maximum concentration Cmax/MIC50 and area under the curve AUC0-24/MIC50 ratios were, respectively, 3.07 ± 0.62 and 7.01 ± 1.03 and 25.48 ± 3.04 and 57.2 ± 3.73 for the 10 and 20 mg/kg doses, respectively. After SC administration of 10 mg/kg, Cmax/MIC50 and AUC0-24/MIC50 ratios were 3.45 ± 0.75 and 33.96 ± 7.46, respectively. After the administration of 10-h pulsed or 24-h continuous medicated water at 20 mg/kg, lower values of Cmax/MIC50 (10-h pulsed: 3.45 ± 0.7; 24-h continuous: 3.05 ± 0.48) and AUC0-24/MIC50 (10-h pulsed: 42.42 ± 6.17; 24-h continuous: 53.32 ± 5.55) were obtained. Based on these results, the European Union-recommended dosage of 10 mg/kg seems ineffective to achieve adequate drug plasma concentrations and even the 20 mg/kg by 10 h pulsed or continuous medicated water administration did not reach completely efficacious concentrations in plasma against colibacillosis. Although the results obtained were not completely encouraging, the medicated water should preferably be provided continuously. To conclude about the efficacy of ENRO treatment against colibacillosis, target tissue concentration should be extensively considered.

Flumequine (FLU) is used in the treatment of systemic bacterial infections in poultry, including colibacillosis, which is a common disease in turkeys. The pharmacokinetic (PK) behavior of FLU administered to 32 healthy turkeys as an oral bolus via gavage or as 10-h pulsed administration in drinking water were compared, using the authorized dose of 15 mg/kg and the double dose of 30 mg/kg. The minimum inhibitory concentrations (MIC) of 235 Escherichia coli field strains isolated from poultry were determined for pharmacodynamics (PD) to develop a PK/PD model. Blood samples were collected at established times over 24 h, and the obtained plasma was analyzed using a liquid chromatography tandem mass spectrometry method that was validated in-house. A monocompartmental model and a noncompartmental model were applied to the data to obtain the PK results. For both types of administration and both dosages, the ratios of the maximum concentration (Cmax)/MIC50 and the area under the plasma concentration-time curve (AUC)/MIC50 achieved were considerably lower than the fluoroquinolone breakpoints usually adopted for efficacy. The Cmax/MIC50 and AUC0-24/MIC50 ratios were, respectively, 0.67 ± 0.09 and 4.76 ± 0.48 and 1.18 ± 0.35 and 7.05 ± 2.40 for the 15 and 30 mg/kg bolus doses, respectively. After 10-h pulsed administration of 15 mg/kg, values of Cmax/MIC50, 0.19 ± 0.02 on d 1 and 0.30 ± 0.08 on d 5 of therapy were obtained, the AUC/MIC50 ratios were 2.09 ± 0.29 and 3.22 ± 0.93 on d 1 and 5, respectively. Higher values were obtained with the doubled dose of 30 mg/kg: the Cmax/MIC50 ratios were 0.49 ± 0.11 on d 1 and 0.69 ± 0.18 on d 5; the AUC/MIC50 ratios were 5.15 ± 1.15 and 6.57 ± 1.92 on d 1 and 5, respectively. Based on these results, FLU administration should be adopted when specific diagnostic findings indicate its efficacy, and revising the dosage scheme to comply with the prudent and responsible use of antimicrobials in veterinary medicine is advisable.

Methicillin and multi-drug resistance were investigated in 136 Staphylococcus intermedius strains of canine origin. The large majority of isolates were found to be mecA-negative by polymerase chain reaction, whereas only four strains were positive for the mecA gene. All mecA-positive strains were confirmed as methicillin-resistant by complementary tests, except for oxacillin disk diffusion, which yielded one false-negative result. A significantly higher resistance to fusidic acid, lincosamides, and cotrimoxazole was observed in methicillin-resistant S. intermedius (MRSI) compared with methicillin-susceptible strains. Although the prevalence of MRSI in dogs appeared to be low, methicillin resistance was confirmed to be associated with multi-drug resistance, suggesting the importance of antimicrobial susceptibility testing of canine S. intermedius strains.

After birth, infant formulas constitute an important or often sole food source for infants during the first months of life. In this study, a survey on the presence of aflatoxin M1 (AFM1) and ochratoxin A (OTA) in the 14 leading brands of infant formulas marketed in Italy was conducted. Mycotoxins were determined by immunoaffinity column clean-up and high-performance liquid chromatography (HPLC) with fluorescence detection. AFM1 was found in two of 185 samples, but at levels below the European legislation limit of 25 ng l(-1). OTA was detected in 133 (72%) samples (range = 35.1-689.5 ng l(-1)). It has been observed that OTA contamination was 80% in the ready-to-use preparations and 63% in the powdered samples. The Scientific Committee for Food (SCF) reviewed the toxicology on OTA and concluded that it would be prudent to reduce exposure to OTA ensuring that exposure is towards the lower end of the range of tolerable daily intakes of 1.2-14 ng kg(-1) body weight day(-1). OTA was also evaluated by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) and a provisional tolerable weekly intake (PTWI) of 100 ng kg(-1) body weight was established. The OTA levels in pre-term ready-to-use infant formulas were sufficient to cause a higher OTA intake than the suggested TDI. The results point out the need to perform controls for prevention programmes especially when attempting to identify risk markers of the infant feed quality.