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To investigate the haemodynamic effects of gamma-hydroxybutyric acid (GHB) in isoflurane-anaesthetized pigs. Experimental, randomized, nonblinded, crossover study. A group of six stress-resistant Landrace pigs (approximately 3 months old; three male, three female; bodyweight 39.2 ± 4 kg, mean ± standard deviation). After premedication (midazolam 0.5 mg kg–1 and ketamine 10 mg kg–1 intramuscularly) and induction [propofol 0.25–0.5 mg kg–1 intravenously (IV)], anaesthesia was maintained with isoflurane in oxygen, and either GHB 250 mg kg–1 IV or an equal volume of saline was administered (minimum washout period 1 week). Systolic (SAP), diastolic (DAP) and mean (MAP) arterial pressures, heart rate and rhythm and respiratory rate were recorded every 5 minutes for 2 hours. Arterial samples were collected for blood gas and pharmacokinetic analyses. Relative changes from baseline were calculated and compared between treatments using a mixed model with time, period and treatment as variables (α < 0.05). Changes from baseline differed significantly between treatments (p < 0.001) for SAP (GHB –1.6 ± 10.7; saline –5.9 ± 14.8 mmHg), DAP (GHB +2.9 ± 9.6; saline –6.5 ± 10.7 mmHg) and MAP (GHB +2.2 ± 10.5; saline –5.7 ± 9.6 mmHg). Statistical analysis of secondary outcomes suggested effects on PaO 2 [GHB –45.2 ± 29.8 mmHg (–6.03 ± 3.97 kPa); saline +24.5 ± 32.4 mmHg (+3.27 ± 4.32 kPa); p < 0.001] and PaCO 2 [GHB –2 ± 10 mmHg (–0.27 ± 1.33 kPa); saline –9 ± 8 mmHg (–1.20 ± 1.07 kPa); p < 0.001]. Mean maximum blood concentration of GHB was 1171.1 ± 229.3 μg mL–1, with volume of distribution 335.3 ± 68.5 mL kg–1, clearance 77.2 ± 19.12 mL kg–1 hour–1 and elimination half-life 3.10 ± 0.80 hours. GHB did not cause severe physiological side effects and may reduce cardiovascular depression. [ABSTRACT FROM AUTHOR]

Sedative as well as protective effects during hypoxia have been described for gamma‐hydroxybutyric acid (GHB). Six swine (Sus scrofa domesticus) of 6 weeks old were administered NaGHB at a dose of 500 mg/kg intravenously (IV) and 500 and 750 mg/kg orally (PO) in a triple cross‐over design. Repeated blood sampling was performed to allow pharmacokinetic analysis of GHB. Whole blood concentration at time point 0 after IV administration was 1727.21 ± 280.73 μg/mL, with a volume of distribution of 339.45 ± 51.41 mL/kg and clearance of 164.94 ± 47.05 mL/(kg h). The mean peak plasma concentrations after PO administration were 326.57 ± 36.70 and 488.01 ± 154.62 μg/mL for 500 mg/kg and 750 mg/kg, respectively. These were recorded at 1.42 ± 0.72 and 1.58 ± 0.58 h after PO dose for GHB 500 mg/kg and 750 mg/kg, respectively. The elimination half‐life for IV and PO 500 mg/kg and PO 750 mg/kg dose was respectively 1.33 ± 0.30, 1.16 ± 0.31 and 1.11 ± 0.33 h. The bioavailability (F) for PO administration was 45%. No clinical adverse effects were observed after PO administration. Deep sleep was seen in one animal after IV administration, other animals showed head pressing and ataxia. [ABSTRACT FROM AUTHOR]

Age-related differences in toxicokinetic processes of deoxynivalenol (DON) and deoxynivalenol-3-glucoside (DON3G) were studied. DON3G [55.7 µg/kg bodyweight (BW)] and an equimolar dose of DON (36 µg/kg BW) were administered to weaned piglets (4 weeks old) by single intravenous and oral administration in a double two-way cross-over design. Systemic and portal blood was sampled at different time points pre- and post-administration and plasma concentrations of DON, DON3G and their metabolites were quantified using validated liquid chromatography-tandem mass spectrometry (LC–MS/MS) and liquid chromatography–high-resolution mass spectrometry (LC–HRMS) methods. Data were processed using tailor-made compartmental toxicokinetic (TK) models to accurately estimate TK parameters. Results were statistically compared to data obtained in a previous study on 11-week-old pigs using identical experimental conditions. Significant age-related differences in intestinal and systemic exposure to both DON and DON3G were noted. Most remarkably, a significant difference was found for the absorbed fraction of DON3G, after presystemic hydrolysis to DON, in weaned piglets compared to 11-week-old piglets (83% vs 16%, respectively), assumed to be mainly attributed to the higher intestinal permeability of weaned piglets. Other differences in TK parameters could be assigned to a higher water/fat body ratio and longer gastrointestinal transit time of weaned piglets. Results may further refine current risk assessment concerning DON and DON3G in animals. Additionally, since piglets possibly serve as a human paediatric surrogate model, results may be extrapolated to human infants. [ABSTRACT FROM AUTHOR]

Intranasal ketamine has recently gained interest in human medicine, not only for its sedative, anaesthetic or analgesic properties, but also in the management of treatment resistant depression, where it has been shown to be an effective, fast acting alternative treatment. Since several similarities are reported between human psychiatric disorders and canine anxiety disorders, intranasal ketamine could serve as an alternative treatment for anxiety disordered dogs. However, to the authors knowledge, intranasal administration of ketamine and its pharmacokinetics have never been described in dogs. Therefore, this study aimed to examine the pharmacokinetics, absolute bioavailability and tolerability of intranasal ketamine administration compared with intravenous administration. Seven healthy, adult laboratory Beagle dogs were included in this randomized crossover study. The dogs received 2 mg/kg body weight ketamine intravenously (IV) or intranasally (IN), with a two-week wash-out period. Prior to ketamine administration, dogs were sedated intramuscularly with dexmedetomidine. Venous blood samples were collected at fixed times until 480 min post-administration and ketamine plasma concentrations were determined by liquid chromatography-tandem mass spectrometry. Cardiovascular parameters and sedation scores were recorded at the same time points. Non-compartmental pharmacokinetic analysis revealed a rapid (Tmax = 0.25 ± 0.14 h) and complete IN bioavailability (F = 147.65 ± 49.97%). Elimination half-life was similar between both administration routes (T1/2el IV = 1.47 ± 0.24 h, T1/2el IN = 1.50 ± 0.97 h). Heart rate and sedation scores were significantly higher at 5 and 10 min following IV administration compared to IN administration, but not at the later time-points. [ABSTRACT FROM AUTHOR]

To investigate the pharmacokinetics and absolute oral bioavailability of meloxicam in guinea pigs. Prospective crossover study. A group of six healthy male Dunkin Hartley guinea pigs. A single dose of meloxicam (1.5 mg kg−1) was administered orally and intravenously (IV) to six healthy male guinea pigs. A wash-out period of 48 hours was taken into account between administrations (oral and IV) in the same animal. Blood was sampled through a central venous catheter before administration (t = 0 hours) and at 0.5, 1, 2, 3, 4, 6, 8, 10, 12, 16, 20, 24 and 28 hours post administration. After centrifugation, plasma concentrations of meloxicam were measured by high-performance liquid chromatography with UV detection, and pharmacokinetic parameters were calculated using noncompartmental analysis. Meloxicam in guinea pigs exhibited a moderate absorption rate after oral dosing (time to maximal plasma concentration 3.7 ± 1.7 hours) and maximal plasma concentration was 0.92 ± 0.30 μg mL−1. After IV administration, total body clearance and volume of distribution were 0.13 ± 0.04 and 0.72 ± 0.36 L kg−1, respectively. Terminal half-life was 3.7 ± 0.7 hours and 3.5 ± 1.1 hours after IV and oral administration, respectively. Body extraction ratio was 0.0087 and mean absorption time was 3.8 ± 1.7 hours. The absolute oral bioavailability was 0.54 ± 0.14 in unfasted guinea pigs. This study reported the pharmacokinetics of meloxicam in guinea pigs. Studies concerning efficacy and safety are the next step towards a rational use of this drug in guinea pigs. [ABSTRACT FROM AUTHOR]

The pharmacokinetics and residue depletion of doxycycline (DOX) in striped catfish (Pagasianodon hypophthalmus) after oral dosage were investigated. The pharmacokinetic experiment was conducted in an aquarium, while the experiment of residue depletion was performed in both an aquarium and earth ponds. Medicated feed was administered orally using the gavage method at a dosage of 20 mg/kg body weight. Blood, liver, and kidney from medicated fish samples were collected. In the depletion experiments, fish were fed medicated feed for five consecutive days at a dosage of 20 mg/kg body weight, with samples collected during and after medication. The concentrations of DOX were quantified using an LC–MS/MS system. The pharmacokinetics parameters of DOX in striped catfish included the absorption rate constant (ka), absorption half‐life (T1/2abs), maximal plasma concentration (Cmax), time to maximal plasma concentration (Tmax), and area under the plasma concentration–time curve from time 0 to 96 h (AUC0–96 h) which were 0.12 h−1, 5.68 h, 1123.45 ng/mL, 8.19 h, and 25,018 ng/mL/h, respectively. Residue depletion results indicated that the withdrawal times of DOX in muscle (with skin) from fish kept in the aquarium were slightly longer than that in fish raised in earth ponds, corresponding to 194 degree‐days compared with 150 degree‐days. In conclusion, administration of DOX at the dosage of 20 mg/kg body weight can be used for treatment of bacterial infections in striped catfish, and a withdrawal time of 5 days at 29.4°C will ensure consumer food safety due to the rapid depletion of DOX from muscle and skin. [ABSTRACT FROM AUTHOR]

A high-performance liquid chromatography method coupled to tandem mass spectrometry was validated in order to study the pharmacokinetics of cefotaxime in shrimp hepatopancreases and plasma, as well as its withdrawal time related to a maximum residue limit (MRL) in shrimp muscle. Pharmacokinetics parameters were investigated through oral medication at a single dose of 25 mg/kg shrimp body weight and subsequent hepatopancreas and plasma cefotaxime concentration measurements at 0.5, 1, 2, 4, 8, 12 and 24 h after shrimp were fed with medication. The maximum concentration of cefotaxime was observed after one hour in the hepatopancreas (Cmax, 19.45 ± 2.10 mg/kg) and 4 h in plasma (0.184 ± 0.061 mg/L). Based on a minimum inhibitory concentration (MIC) of cefotaxime of 4.13 mg/L against Vibrio parahaemolyticus (known to cause acute hepatopancreatic necrosis disease (AHPND) in white leg shrimp), it was observed that the time during which the hepatopancreas cefotaxime concentration was above the MIC was 23 h. An every 24 h cefotaxime treatment could thus be effective in fighting against this bacterium in shrimp. The withdrawal time of cefotaxime was determined after shrimp were fed with medicated feed once a day and twice a day for three consecutive days. Shrimp muscle was collected on day 1 and day 3 during medication and 1, 3, 7, 14 and 21 days after medication was stopped. Considering an MRL of 50 μg/kg, the withdrawal times were 8.5 degree-days (corresponding to 6.9 h at 29.5 °C) after shrimp were fed with medicated feed once a day for 3 days and 95.5 degree-days (77.7 h at 29.5 °C) after shrimp were fed with medicated feed twice a day for 3 days. Moreover, histological analysis revealed that feeding shrimp with cefotaxime at the given dose in once- or twice-a-day treatments did not negatively impact the shrimp hepatopancreas. [ABSTRACT FROM AUTHOR]

• Microdialysis was safe and feasible over multiple days in septic piglets and children with sepsis. • Sepsis impacted the relative recovery. • Comprehensive method validation is required in the target study population. Knowledge on the tissue penetration of piperacillin-tazobactam in children with sepsis is lacking. In this study, the feasibility and performance of microdialysis experiments were explored in septic piglets and children as part of a translational research project. Multiple-day microdialysis investigations were performed in muscle tissue of 22 piglets (of which 11 were septic) and 6 children with sepsis. An in vitro experiment preceded the (pre)clinical trials to derive optimal experimental settings and calibration technique. Linear mixed-effects models quantified the impact of sepsis on relative recovery (RR) and intercatheter, interindividual, interoccasion, and residual variability. In vivo microdialysis was well tolerated in piglets and children, with no significant adverse events reported. Using identical experimental settings, lower RR values were recorded in healthy and septic piglets (range: piperacillin, 17.2–29.1% and tazobactam, 23.5–29.1%) compared with the in vitro experiment (piperacillin, 43.3% and tazobactam, 55.3%), and there were unacceptably low values in children with sepsis (<10%). As a result, methodological changes were made in the pediatric trial. Realistic tissue concentration-time curves were derived in piglets and children. In piglets, sepsis reduced the RR. The greatest contributors to RR variability were residual (>40%) and interoccasion (>30%) variability. The internal standard method was the preferred calibration technique in both piglets and children. Microdialysis is a safe and applicable method for the measurement of tissue drug concentrations in piglets and children. This study demonstrated the impact of experimental settings, sepsis, and target population on individual RR. [ABSTRACT FROM AUTHOR]

The aims of this study were (i) to determine the effect of an algoclay-based decontaminant on the oral availability of three mycotoxins (deoxynivalenol; DON, ochratoxin A; OTA, and aflatoxin B1; AFB1) using an oral bolus model and (ii) to determine the effect of this decontaminant on the performance, intestinal morphology, liver oxidative stress, and metabolism, in broiler chickens fed a diet naturally contaminated with DON. In experiment 1, sixteen 27-day-old male chickens (approximately 1.6 kg body weight; BW) were fasted for 12 h and then given a bolus containing either the mycotoxins (0.5 mg DON/kg BW, 0.25 mg OTA/kg BW, and 2.0 mg AFB1/kg BW) alone (n = 8) or combined with the decontaminant (2.5 g decontaminant/kg feed; circa 240 mg/kg BW) (n = 8). Blood samples were taken between 0 h (before bolus administration) and 24 h post-administration for DON-3-sulphate, OTA, and AFB1 quantification in plasma. The algoclay decontaminant decreased the relative oral bioavailability of DON (39.9%), OTA (44.3%), and AFB1 (64.1%). In experiment 2, one-day-old male Ross broilers (n = 600) were divided into three treatments with ten replicates. Each replicate was a pen with 20 birds. The broiler chickens were fed a control diet with negligible levels of DON (0.19–0.25 mg/kg) or diets naturally contaminated with moderate levels of DON (2.60–2.91 mg/kg), either supplemented or not with an algoclay-based decontaminant (2 g/kg diet). Jejunum villus damage was observed on day 28, followed by villus shortening on d37 in broiler chickens fed the DON-contaminated diet. This negative effect was not observed when the DON-contaminated diet was supplemented with the algoclay-based decontaminant. On d37, the mRNA expression of glutathione synthetase was significantly increased in the liver of broiler chickens fed the DON-contaminated diet. However, its expression was similar to the control when the birds were fed the DON-contaminated diet supplemented with the algoclay-based decontaminant. In conclusion, the algoclay-based decontaminant reduced the systemic exposure of broiler chickens to DON, OTA, and AFB1 in a single oral bolus model. This can be attributed to the binding of the mycotoxins in the gastrointestinal tract. Moreover, dietary contamination with DON at levels between 2.69 and 2.91 mg/kg did not impair production performance but had a negative impact on broiler chicken intestinal morphology and the liver redox system. When the algoclay-based decontaminant was added to the diet, the harm caused by DON was no longer observed. This correlates with the results obtained in the toxicokinetic assay and can be attributed to a decreased absorption of DON. [ABSTRACT FROM AUTHOR]

Microsomes are an ideal medium to investigate cytochrome P450 (CYP450) enzyme-mediated drug metabolism. However, before microsomes are prepared, tissues can be stored for a long time. Studies about the stability of these enzymes in porcine hepatic and intestinal tissues upon storage are lacking. To be able to investigate CYP450 stability in microsomes prepared from these tissues, a highly sensitive and rapid HPLC-MS/MS method for the simultaneous determination of six CYP450 metabolites in incubation medium was developed and validated. The metabolites, paracetamol (CYP1A), 7-hydroxy-coumarin (CYP2A), 1-hydroxy-midazolam (CYP3A), 4-hydroxy-tolbutamide (CYP2C), dextrorphan (CYP2D), and 6-hydroxy-chlorzoxazone (CYP2E) were extracted with ethyl acetate at pH 1.0, followed by evaporation and separation on an Agilent Zorbax Eclipse Plus C18 column. The method was fully validated in a GLP-compliant laboratory according to European guidelines and was highly sensitive (LOQ = 0.25–2.5 ng/mL), selective, had good precision (RSD-within, 1.0–9.1%; RSD-between, 1.0–18.4%) and accuracy (within-run, 83.3–102%; between-run, 78.5–102%), and showed no relative signal suppression and enhancement. Consequently, this method was applied to study the stability of porcine hepatic and intestinal CYP450 isoenzymes when tissues were stored at − 80 °C. The results indicate that porcine CYP450 isoenzymes are stable in tissues at least up to 4 months when snap frozen and stored at − 80 °C. Moreover, the results indicate differences in porcine CYP450 stability compared to rat, rabbit, and fish CYP450, as observed by other research groups, hence stressing the importance to investigate the CYP450 stability of a specific species. [ABSTRACT FROM AUTHOR]

Cytochrome P450 (CYP450) drug biotransformation enzymes and multidrug resistance (MDR) proteins may influence drug disposition processes. The first part of the study aimed to evaluate the effect of mycotoxins deoxynivalenol (DON) and/or fumonisins (FBs), at contamination levels approaching European Union guidance levels, on intestinal and hepatic CYP450 enzymes and MDR proteins gene expression in broiler chickens. mRNA expression of genes encoding CYP450 enzymes (CYP3A37, CYP1A4 and CYP1A5) and drug transporters (MDR1/ABCB1 and MRP2/ABCC2) was determined using qRT-PCR. A significant up-regulation of CYP1A4 ( P = 0.037 ) and MDR1 ( P = 0.036 ) was observed in the jejunum of chickens fed a diet contaminated with FBs. The second part of this study aimed to investigate the impact of feeding a FBs contaminated diet on the oral absorption of enrofloxacin (10 mg/kg BW), a MDR1 substrate. A significant (P = 0.045), however small, decreased area under the plasma concentration-time curve (AUC 0-48 h, mean ± SD) was observed for enrofloxacin in chickens fed the FBs contaminated diet compared to the control group, 16.28 ± 1.82 h μg/mL versus 18.27 ± 1.79 h μg/mL. These findings suggest that concurrent administration of drugs with FBs contaminated feed might alter the pharmacokinetic characteristics of CYP1A4 substrate drugs and MDR1 substrates, such as enrofloxacin. [ABSTRACT FROM AUTHOR]

Ornithobacterium rhinotrachealeis an avian respiratory pathogen that affects turkeys. The objective of this study was to evaluate the clinical efficacy of gamithromycin (GAM) againstO. rhinotrachealein turkeys. The birds were inoculated oculonasally with 108colony-forming units (cfu) ofO. rhinotracheale, preceded by infection with avian metapneumovirus. In addition to a negative (CONTR−) and a positive control group (CONTR+) there were two treated groups administered GAM (6 mg/kg) either subcutaneously (GAM SC) or orally (GAM PO) by administration as a single bolus at one-day post-bacterial infection (p.b.i.). From the start of the avian metapneumovirus infection until the end of the experiment, the turkeys were examined clinically and scored daily. In addition, tracheal swabs were collected at several days p.b.i. Necropsy was performed at 4, 8 and 12 days p.b.i. to evaluate the presence of gross lesions, and to collect trachea and lung tissue samples and air sac swabs forO. rhinotrachealequantification. The clinical score of the GAM SC group showed slightly lower values and birds recovered earlier than those in the GAM PO and CONTR+ groups.O. rhinotrachealecfus were significantly reduced in tracheal swabs of the SC group between 2 and 4 days p.b.i. At necropsy, CONTR+ showed higherO. rhinotrachealecfu in lung tissues compared to the treated groups. Moreover, at 8 days p.b.i. only the lung samples of CONTR+ were positive. In conclusion, the efficacy of GAM againstO. rhinotrachealewas demonstrated, especially in the lung tissue. However, the PO bolus administration of the commercially available product was not as efficacious as the SC bolus. [ABSTRACT FROM AUTHOR]

The gastrointestinal tract is the first target after ingestion of the mycotoxin deoxynivalenol (DON) via feed and food. Deoxynivalenol is known to affect the proliferation and viability of animal and human intestinal epithelial cells. In addition to DON, feed and food is often co-contaminated with modified forms of DON, such as 3-acetyldeoxynivalenol (3ADON), 15-acetyl-deoxynivalenol (15ADON) and deoxynivalenol-3-β-D-glucoside (DON3G). The goal of this study was to determine the in vitro intrinsic cytotoxicity of these modified forms towards differentiated and proliferative porcine intestinal epithelial cells by means of flow cytometry. Cell death was assessed by dual staining with Annexin-V-fluorescein isothiocyanate (FITC) and propidium iodide (PI), which allows the discrimination of viable (FITC−/PI−), apoptotic (FITC+/PI−) and necrotic cells (FITC+/PI+). Based on the data from the presented pilot in vitro study, it is concluded that cytotoxicity for proliferative cells can be ranked as follows: DON3G ≪ 3ADON < DON ≈ 15ADON. [ABSTRACT FROM AUTHOR]

The macrolide gamithromycin (GAM) has the ability to accumulate in tissues of the respiratory tract. Consequently, GAM might be a suitable antibiotic to treat bacterial respiratory infections in poultry, such as Ornithobacterium rhinotracheale. As O. rhinotracheale infections are common in turkey flocks, the aim of this study was to determine the pharmacokinetic (PK) parameters of GAM in plasma, lung tissue, and pulmonary epithelial lining fluid (PELF) of turkeys and to correlate them with pharmacodynamic (PD) characteristics (PK/PD). The animal experiment was performed with 64 turkeys, which received either a subcutaneous (SC, n = 32) or an oral (PO, n = 32) bolus of 6 mg GAM/kg body weight (BW). GAM concentrations in plasma, lung tissue, and PELF were measured at different time points post administration (p.a.), and PK characteristics were determined using non-compartmental modeling. The maximum plasma concentration after PO administration was ten-fold lower than after SC injection (0.087 and 0.89 µg/mL, respectively), whereas there was no difference in lung concentrations between both routes of administration. However, lung concentrations at day 1 p.a. were significantly higher than plasma levels for both routes of administration (2.22 and 3.66 µg/g for PO and SC, respectively). Consequently, lung/plasma ratios were high, up to 50 and 80 after PO and SC administration, respectively. GAM could not be detected in PELF, although this might be attributed to the collection method of PELF in birds. The GAM minimum inhibitory concentration (MIC) was determined for 38 O. rhinotracheale strains; MIC50 and MIC90 were 2 and >32 µg/mL, respectively. PK/PD correlation for lung tissue demonstrated that the time above the MIC90 of the susceptible population (2 µg/mL) was 1 day after PO bolus and 3.5 days after SC administration. The area under the curve (AUClast)/MIC ratios for lung tissue after SC and PO administration were 233 and 90, respectively. To conclude, GAM is highly distributed to lung tissue in turkey poults, suggesting that it has the potential to be used to treat respiratory infections such as O. rhinotracheale. [ABSTRACT FROM AUTHOR]

The aim of present study was to develop a rapid and reliable analytical method for quantification of ketamine and its major metabolite, norketamine, in bovine plasma. A cost-effective and high-throughput extraction procedure was combined with a sensitive and specific liquid chromatography-heated electrospray ionization-tandem mass spectrometry method. Sample preparation consisted of a protein precipitation step using methanol followed by a clean-up using a HybridSPE-phospholipid column and further dilution by a factor 1/10 with water before injection onto the LC-MS/MS instrument. A gradient elution program was performed with 1 % formic acid in water and 0.1 % acetic acid in methanol as mobile phases. Ionization was performed by a heated electrospray ionization (h-ESI) probe operating in the positive ionization mode. Following transitions were monitored in the selected reaction monitoring mode: 238.0 > 124.9/220.0 for ketamine and 224.0 > 207.0/124.9 for norketamine. The method was validated in-house. Matrix-matched calibration graphs were prepared for ketamine and norketamine and correlation and goodness-of-fit coefficients were 0.9991 and 0.9997 and 7.7 and 4.0 %, respectively. The within- and between-run precision and accuracy were evaluated and the results fell within the ranges specified. The limit of quantification was 0.1 µg/mL for both analytes, whereas limits of detection were 9.6 and 3.5 ng/mL for ketamine and norketamine, respectively. The method was applied on biological samples from a pharmacokinetic study in calves and demonstrated the suitability of the method for this application. [ABSTRACT FROM AUTHOR]

The use of chemotherapeutics as a possible treatment strategy in avian oncology is steadily increasing over the last years. Despite this, literature reports regarding dosing strategies and pharmacokinetic behaviour of chemotherapeutics in avian species are lacking. The aim of the present study was to investigate the pharmacokinetics of carboplatin in a representative species of the order of Galliformes, Anseriformes, Columbiformes and Psittaciformes. Eight chickens, ducks and pigeons and twenty-eight parakeets were administered carboplatin intravenously (5 mg/kg body weight). A specific and sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for quantification of the free carboplatin in plasma of the four birds species (limit of quantification: 20 ng/mL for chicken and duck, 50 ng/mL for pigeon and 100 ng/mL for parakeets). Non-compartmental pharmacokinetic analysis and allometric scaling demonstrated a significant correlation (R² = 0.9769) between body weight (BW) and elimination half-life (T1/2el). T1/2el ranged from 0.41 h in parakeets (BW: 61 ± 8 g) to 1.16 h chickens (BW: 1909 ± 619 g). T1/2el is a good parameter for dose optimization of carboplatin in other avian species, since also the previously reported T1/2el in cockatoos (average BW: 769 ± 68 g) of 1.00 h corresponds to the results obtained in the present study. [ABSTRACT FROM AUTHOR]

A sensitive and specific method for the quantitative determination of gamithromycin in animal plasma, lung tissue and pulmonary epithelial lining fluid (PELF) using liquid chromatography combined with heated electrospray ionization tandem mass spectrometry (LC–MS/MS) was developed. The sample preparation was rapid, straightforward and consisted of a deproteinization and phospholipid removal step using an Oasis ® Ostro™ 96-well plate (chicken, turkey and calf plasma) or HybridSPE ® -Phospholipid SPE cartridges (pig plasma and turkey lung tissue), while a liquid–liquid extraction with diethyl ether in alkaline medium was used for PELF of turkey poults. Chromatography was performed on a C18 Hypersil GOLD column using 0.01 M ammonium acetate in water with a pH of 9, and acetonitrile as mobile phases. The MS/MS instrument was operated in the positive electrospray ionization mode and the following selected reaction monitoring transitions were monitored for gamithromycin (protonated molecule > product ion): m / z 777.45 > 619.35 and m / z 777.45 > 157.80 for quantification and identification, respectively. The method was validated in-house: matrix-matched calibration graphs were prepared and good linearity ( r ≥ 0.99) was achieved over the concentration ranges tested (2.5–10,000 ng mL −1 for chicken, pig and calf plasma; 5.0–2500 ng mL −1 for turkey plasma; 50–10,000 ng g −1 for turkey lung tissue and 20–1000 ng mL −1 for turkey PELF). Limits of quantification (LOQ) were 2.5 ng mL −1 for chicken, pig and calf plasma and 5.0 ng mL −1 for turkey plasma, while the limits of detection (LOD) ranged between 0.007 and 0.07 ng mL −1 . For lung tissue and PELF, respective LOQ and LOD values of 50 ng g −1 and 0.76 ng g −1 (lung tissue) and 20 ng mL −1 and 0.1 ng mL −1 (PELF) were obtained. The results for the within-day and between-day precision, expressed as relative standard deviation (RSD), fell within the maximal RSD values. The accuracy fell within −30% to +10% (concentrations 1–10 ng mL −1 ) or −20% to +10% (concentrations > 10 ng mL −1 or ng g −1 ) of the theoretical concentration. The method was successfully applied for the quantitative determination of gamithromycin in plasma samples of chickens, turkeys, pigs and calves; and in lung tissues and PELF of turkeys, all derived from pharmacokinetic studies in these animal species. [ABSTRACT FROM AUTHOR]

For the first time, a sensitive and specific method was developed and fully validated for the quantification of the EU marker residue of tiamulin, 8-α-hydroxy-mutilin, in rabbit muscle and liver tissues using liquid chromatography combined with positive heated electrospray ionization triple quadrupole mass spectrometry. The mass spectrometer was operated in the selected reaction monitoring (SRM) mode with selection of the [M + H] ion in both quadrupoles 1 and 3, resulting in the SRM transition m/z 337.25 > 337.25 for quantification. Chromatography was performed using a Hypersil Gold C18 column using a gradient elution program with water and methanol as mobile phases. The sample preparation procedure for the analysis of 8-α-hydroxy-mutilin in liver and muscle samples consisted of three main steps: (1) extraction of the tissue matrix using 0.1 N hydrochloric acid/acetone (50/50, v/v), (2) hydrolysis of tiamulin and metabolites to 8-α-hydroxy-mutilin in alkaline medium at 45 °C, and (3) liquid-liquid extraction in acidic medium using ethyl acetate. This is the first method presenting fully validated results, encompassing a linearity of 50 to 2,000 μg/kg, within-run and between-run accuracy and precision, limit of quantification (50 μg/kg for both muscle and liver tissues), limit of detection (muscle, 11.9 μg/kg; liver, 20.6 μg/kg), extraction recovery (muscle, 66.2 %; liver, 75.5 %), signal suppression and enhancement (muscle, 51.7 %; liver, 43.3 %), carryover, applicability and practicability, and stability during storage and analysis. This novel method is therefore sensitive enough to be used for residue depletion studies of tiamulin in rabbits and for food safety monitoring with respect to MRL compliance of residues. [ABSTRACT FROM AUTHOR]

Modified mycotoxins formed by plants, fungi and during some food processing steps may remain undetected by analytical methods, potentially causing underestimation of mycotoxin exposure and risk. Furthermore, due to altered physico-chemical characteristics of modified mycotoxins, these compounds might have different gastro-intestinal absorption compared to the unmodified forms, leading to altered modified mycotoxin plasma concentrations. Additionally, modified mycotoxins can be converted back into their corresponding unmodified forms by in vivo hydrolysis upon oral ingestion. This review aims to describe the current knowledge on the production, occurrence, toxicity and toxicokinetic properties of the modified Fusarium mycotoxins. The need for more occurrence data to correctly assess the risks associated with these modified mycotoxins is clearly indicated, including differences between commodities as well as geographical and climatological influences. Research on toxicity of these modified forms demonstrates the possibility of significant decreases as well as increases in the toxic effects of these compounds compared with those of the unmodified forms. Their toxicokinetics demonstrates that a decreased (increased) polarity of modified mycotoxins might cause enhanced (decreased) oral absorption. The possibility of in vivo hydrolysis, altered toxicity and their wide-spread occurrence makes modified mycotoxins a complex threat for which a risk assessment will require prospective multi-disciplinary efforts. [ABSTRACT FROM AUTHOR]

Both deoxynivalenol (DON) and fumonisin B1 (FB1) are common contaminants of feed. Fumonisins (FBs) in general have a very limited oral bioavailability in healthy animals. Previous studies have demonstrated that chronic exposure to DON impairs the intestinal barrier function and integrity, by affecting the intestinal surface area and function of the tight junctions. This might influence the oral bioavailability of FB1, and possibly lead to altered toxicity of this mycotoxin. A toxicokinetic study was performed with two groups of 6 broiler chickens, which were all administered an oral bolus of 2.5 mg FBs/kg BW after three-week exposure to either uncontaminated feed (group 1) or feed contaminated with 3.12 mg DON/kg feed (group 2). No significant differences in toxicokinetic parameters of FB1 could be demonstrated between the groups. Also, no increased or decreased body exposure to FB1 was observed, since the relative oral bioavailability of FB1 after chronic DON exposure was 92.2%. [ABSTRACT FROM AUTHOR]

The aim of this study was to characterize 27 feed additives marketed as mycotoxin binders and to screen them for their in vitro zearalenone (ZEN) adsorption. Firstly, 27 mycotoxin binders, commercially available in Belgium and The Netherlands, were selected and characterized. Characterization was comprised of X-ray diffraction (XRD) profiling of the mineral content and d-spacing, determination of the cation exchange capacity (CEC) and the exchangeable base cations, acidity, mineral fraction, relative humidity (RH) and swelling volume. Secondly, an in vitro screening experiment was performed to evaluate the adsorption of a single concentration of ZEN in a ZEN:binder ratio of 1:20,000. The free concentration of ZEN was measured after 4 h of incubation with each of the 27 mycotoxin binders at a pH of 2.5, 6.5 and 8.0. A significant correlation between the free concentration of ZEN and both the d-spacing and mineral fraction of the mycotoxin binders was seen at the three pH levels. A low free concentration of ZEN was demonstrated using binders containing mixed-layered smectites and binders containing humic acids. [ABSTRACT FROM AUTHOR]

Background: The (mis)use of fluoroquinolones in the fowl industry has led to an alarming incidence of fluoroquinolone resistance in pathogenic as well as commensal bacteria. Next to simply reducing antimicrobial consumption, optimizing dosage regimens can be regarded as a suitable strategy to reduce antimicrobial resistance development without jeopardizing therapy efficacy and outcome. A first step in order to limit antimicrobial resistance is to assess the exposure of the intestinal microbiota to enrofloxacin after different treatment strategies. Therefore, a study was conducted in broiler chickens to assess the effect of route of administration (oral versus intramuscular) and dose escalation (10 and 50 mg/kg body weight) on plasma and intestinal concentrations of enrofloxacin and its main metabolite ciprofloxacin after treatment with enrofloxacin once daily for five consecutive days. Four different parts of the intestinal tract were sampled: ileum, cecum, colon and cloaca. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to quantify both analytes in plasma and intestinal content. Sample preparation prior to LC-MS/MS analysis consisted of extraction with ethyl acetate. For intestinal content samples PBS buffer was added before extraction. The supernatant was evaporated to dryness and resuspended in water prior to analysis. Results: The results in plasma and intestinal content demonstrated that biotransformation of enro- to ciprofloxacin in broiler chickens is limited. In general, the intestinal microbiota in cecum and colon is exposed to significant levels of enrofloxacin after conventional treatment (21–130 μg/g). A clear increase of intestinal concentrations was demonstrated after administration of a five-fold higher dose (31–454 μg/g). After intramuscular administration, intestinal concentrations were comparable, except for the higher levels in cloaca due to the complete bioavailability and urinary excretion. Conclusions: The intestinal microbiota is exposed to high levels of the antimicrobial, after oral as well as parenteral therapy. Furthermore, a dose and time dependent correlation was observed. The impact of the detected intestinal levels on resistance selection in the intestinal microbiota has to be further investigated. [ABSTRACT FROM AUTHOR]

In order to assess the in vivo efficacy of mycotoxin binders, specific toxicokinetic parameters should be measured according to European guidelines. For this purpose, an absorption model in pigs is described with emphasis on absorption kinetics. Pigs received a single oral bolus of the mycotoxin deoxynivalenol alone or in combination with active carbon (applied as mycotoxin binder). After administration of deoxynivalenol alone, significant plasma amounts of deoxynivalenol were detected and kinetic parameters were calculated using a one compartmental model. Activated carbon completely prevented the absorption of deoxynivalenol as no plasma amounts could be detected. [ABSTRACT FROM AUTHOR]

Highlights: [•] LC–MS/MS method for quantitative determination of closantel in bovine and ovine milk. [•] Optimized extraction and clean-up using Oasis® MAX® SPE columns. [•] Method validation of bovine milk according to EU Volume 8 (2005). [•] Method validation of ovine milk according to EU Volume 8 (2005) and VICH GL49 (2012). [•] Although both guidelines cover similar parameters, significant differences exist. [ABSTRACT FROM AUTHOR]

Abstract: The aim of present study was to reveal the toxicokinetic properties and absolute oral bioavailability of enniatin B1 in pigs. Five pigs were administered this Fusarium mycotoxin per os and intravenously in a two-way cross-over design. The toxicokinetic profile fitted a two-compartmental model. Enniatin B1 is rapidly absorbed after oral administration (T 1/2a =0.15h, T max =0.24h) and rapidly distributed and eliminated as well (T 1/2elα =0.15h; T 1/2elβ =1.57h). The absolute oral bioavailability is high (90.9%), indicating a clear systemic exposure. After intravenous administration, the mycotoxin is distributed and eliminated rapidly (T 1/2elα =0.15h; T 1/2elβ =1.13h), in accordance with oral administration. [Copyright &y& Elsevier]

Highlights: [•] Effects of DON on unspecific and specific parameters in turkey poults was studied. [•] The efficacy of a GMA mycotoxin binder on these parameters was evaluated. [•] GMA could alleviate some negative effects of DON on unspecific parameters. [•] GMA did not prevent intestinal absorption of DON or its metabolite DOM-1. [ABSTRACT FROM AUTHOR]

Abstract: A sensitive and reliable method was developed for the identification and quantification of beauvericin, enniatin A, A1, B and B1 in pig plasma using liquid chromatography combined with heated electrospray ionization tandem mass spectrometry. Sample clean-up consisted of a deproteinization step using acetonitrile, followed by evaporation of the supernatant and resuspension of the dry residue in acetonitrile/water (80/20, v/v). The method was in-house validated: matrix-matched calibration graphs were prepared for all compounds and correlation and goodness-of-fit coefficients ranged between 0.9980 and 0.9995 and between 5.2% and 11.3%, respectively. The within- and between-run precision and accuracy were evaluated and the results fell within the ranges specified. The limits of quantification were 0.1ng/mL for enniatin A and A1 and 0.2ng/mL for beauvericin, enniatin B and B1, whereas limits of detection were≤10pg/mL for all analytes. The method has been applied for the analysis of real plasma samples from one pig that received an oral bolus (0.05mg/kg BW) of the investigated mycotoxins. At the applied dosage, the results indicated the suitability of the method for use in toxicokinetic studies with enniatins. [Copyright &y& Elsevier]

Abstract: An in vitro model simulating the intestinal barrier for efficacy and drug interaction testing of mycotoxin detoxifying agents was developed using Transwell® cell culture inserts. Intestinal porcine epithelial cells derived from the jejunum of piglets were exposed to DON and a mycotoxin binder (efficacy testing) or exposed to tylosin and a mycotoxin binder (drug interaction testing). Active carbon and bentonite were used in the efficacy and drug interaction trials, respectively, to validate the developed model. The evaluated parameters were passage of DON and tylosin through the epithelial monolayer, the integrity of the monolayer by measurements of the trans-epithelial electrical resistance and the viability of the monolayer using the neutral red assay. In the efficacy model it was shown that active carbon effectively bound DON at both non-cytotoxic and cytotoxic concentrations of DON, respectively 0.5 and 1μg/mL. Moreover, the negative effects of DON at cytotoxic concentrations on cellular viability and integrity were completely offset. A commercially available modified gluco-mannan binder was also tested and it was able to partly reduce the negative effects on these latter parameter. Moreover, it reduced the transepithelial passage of DON with 37% to 57% compared to active carbon, at both cytotoxic and non-cytotoxic concentrations of DON. In our drug interaction model, the interaction between tylosin and mycotoxin binders was investigated as some authors suggest binding of macrolide antibiotics to bentonite clays. Indeed, a bentonite clay showed decreased passage of tylosin through the epithelial monolayer, indicating binding of tylosin by bentonite. This indicates that the combined use of bentonite and tylosin in the feed could lead to therapy failure. The modified gluco-mannan binder did not alter the passage of tylosin significantly, indicating safe combined use. [Copyright &y& Elsevier]

Abstract: Mycotoxins lead to economic losses in animal production. A way to counteract mycotoxicosis is the use of detoxifiers. The European Food Safety Authority stated that the efficacy of detoxifiers should be investigated based on toxicokinetic studies. Little information is available on the absolute oral bioavailability and the toxicokinetic parameters of deoxynivalenol, T-2 and zearalenone in broilers. Toxins were administered intravenously and orally in a two-way cross-over design. For deoxynivalenol a bolus of 0.75mg/kg BW was administered, for T-2 toxin 0.02mg/kg BW and for zearalenone 0.3mg/kg BW. Blood was collected at several time points. Plasma levels of the mycotoxins and their metabolite(s) were quantified using LC–MS/MS methods and toxicokinetic parameters were analyzed. Deoxynivalenol has a low absolute oral bioavailability (19.3%). For zearalenone and T-2 no plasma levels above the limit of quantification were observed after an oral bolus. Volumes of distribution were recorded, i.e. 4.99, 0.14 and 22.26L/kg for deoxynivalenol, T-2 toxin and zearalenone, respectively. Total body clearance was 0.12, 0.03 and 0.48L/minkg for deoxynivalenol, T-2 toxin and zearalenone, respectively. After IV administration, T-2 toxin had the shortest elimination half-life (3.9min), followed by deoxynivalenol (27.9min) and zearalenone (31.8min). [Copyright &y& Elsevier]

Abstract: A sensitive and reliable multi-mycotoxin method was developed for the identification and quantification of several toxicological important mycotoxins such as deoxynivalenol (DON), deepoxy-deoxynivalenol (DOM-1), T-2 toxin (T-2), HT-2 toxin (HT-2), zearalenone (ZON), zearalanone (ZAN), α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), α-zearalanol (α-ZAL), β-zearalanol (β-ZAL), ochratoxin A (OTA), fumonisin B1 (FB1) and aflatoxin B1 (AFB1) in pig plasma using liquid chromatography combined with heated electrospray ionization triple quadrupole tandem mass spectrometry (LC–h-ESI-MS/MS). Sample clean-up consisted of a deproteinization step using acetonitrile, followed by evaporation of the supernatant and resuspension of the dry residue in water/methanol (85/15, v/v). Each plasma sample was analyzed twice, i.e. once in the ESI+ and ESI− mode, respectively. This method can be used for the assessment of animal exposure to mycotoxins and in the diagnosis of mycotoxicoses. For the performance of toxicokinetic studies with individual mycotoxins, highly sensitive analyte-specific LC–MS/MS methods were developed. The multi-mycotoxin and analyte-specific methods were in-house validated: matrix-matched calibration graphs were prepared for all compounds and correlation and goodness-of-fit coefficients ranged between 0.9974–0.9999 and 2.4–15.5%, respectively. The within- and between-run precision and accuracy were evaluated and the results fell within the ranges specified. The limits of quantification for the multi-mycotoxin and analyte-specific methods ranged from 2 to 10ng/mL and 0.5 to 5ng/mL, respectively, whereas limits of detection fell between 0.01–0.52ng/mL and <0.01–0.15ng/mL, respectively. [Copyright &y& Elsevier]

Florfenicol is commonly used to treat bacterial diseases in striped catfish Pangasianodon hypophthalmus aquaculture with limited knowledge of pharmacokinetic database, explaining treatment failures experienced by farmers. Striped catfish (100 g) were fed medicated feed (10 mg florfenicol/kg fish) by oral gavage. Plasma, liver, and kidney were collected up to 96.0 h after drug administration and analyzed for florfenicol. A single oral gavage dose of 10 mg florfenicol/kg fish revealed a rapid elimination, demonstrated by an elimination half-life (T1/2el) of 2.56 h. Florfenicol amine was found in liver and kidney tissue. The calculation of T > MIC (time that the concentration remains above the minimum inhibitory concentration) revealed a potential therapeutic effect on bacillary necrosis, a common disease in striped catfish if the dosing interval is 12 h. The depletion of florfenicol in striped catfish muscle was determined in feeding trials with medicated feed containing florfenicol at 10 mg/kg fish for five consecutive days in tank and pond conditions. The fast elimination of florfenicol resulted in a withdrawal time of two days (or 39 degree days) in proposed tank-rearing conditions and four days (or 112 degree days) in pond-rearing conditions. [ABSTRACT FROM AUTHOR]

Background: In people and dogs, torasemide has higher bioavailability, longer half‐life, and longer duration of action than equivalent doses of furosemide but data regarding pharmacological properties of torasemide in cats are limited. Objective: To assess pharmacokinetic and pharmacodynamic parameters of torasemide in healthy cats, and to investigate the effects of a single administration of torasemide on indicators of diuresis, plasma creatinine concentration, blood pressure, electrolyte concentrations and markers of the renin‐angiotensin‐aldosterone system (RAAS). Animals: Six clinically healthy adult European shorthair cats. Methods: Randomized 4‐period crossover design with 3 groups and 4 treatments. Pharmacokinetic parameters were obtained using a noncompartmental analysis, and the clinically effective dose was assessed using a Hill model. Results: Mean absolute bioavailability was estimated at 88.1%. Mean total body clearance was 3.64 mL/h/kg and mean terminal half‐life was 12.9 hours. Urine output significantly increased after torasemide administration (P <.001). The urine sodium : potassium ratio (uNa : uK) paralleled and was statistically correlated to urine output (P <.001). Administration of a single torasemide dose led to a significant dose‐dependent increase in urine aldosterone : creatinine ratio (uAldo : C; P <.001) and a transient decrease in plasma potassium concentration (P <.001) but did not affect blood pressure or plasma creatinine concentration. Conclusions and Clinical Importance: A single torasemide dose leads to a significant increase in diuresis and renin‐angiotensin‐aldosterone system (RAAS) activation in healthy cats, with high absolute bioavailability, and without clinically relevant adverse effects. Pharmacokinetic parameters indicate that once daily dosing of 0.27 mg/kg may be appropriate in a clinical setting. [ABSTRACT FROM AUTHOR]

The interaction between bentonite and tylosin was investigated in broiler chickens, based on pharmacokinetic characteristics obtained in vivo. Simultaneous oral administration of bentonite and tylosin significantly lowered plasma levels of tylosin and reduced the area under the plasma concentration-time curve (AUC0-inf), maximal plasma concentration (Cmax), time to maximal plasma concentration (Tmax) and relative oral bioavailability. The results prove unambiguously the binding of tylosin by bentonite. Simultaneous administration of tylosin (in the drinking water or feed) and bentonite (mixed in the feed as a mycotoxin binder) should therefore be avoided. [ABSTRACT FROM AUTHOR]

• Plasma concentrations of imepitoin were lower after rectal than oral administration. • Time to maximal concentration did not differ significantly between administration routes. • This study does not support rectal administration of imepitoin in dogs. The purpose of the present study was to investigate if rectal administration of imepitoin in healthy dogs leads to plasma concentrations comparable to those after oral administration. Significantly lower systemic exposure and maximal plasma concentration (C max) of imepitoin was achieved after rectal compared to oral administration (P ≤ 0.001). Therefore, this study does not support the rectal administration of imepitoin in dogs. [ABSTRACT FROM AUTHOR]

Mycotoxins frequently contaminate food and feed materials, posing a threat to human and animal health. Fusarium species produce important mycotoxins with regard to their occurrence and toxicity, especially deoxynivalenol (DON), fumonisin B1 (FB1), zearalenone (ZEN) and T-2 toxin (T-2). The susceptibility of an animal species towards the effects of these toxins in part depends on the absorption, distribution, metabolism and excretion (ADME processes) of these toxins from the body. For humans, in vivo information is scarce and often animal data is used for extrapolation to humans. From a kinetic and safety point of view, the pig seems to be a promising animal model to aid in the assessment of the toxicological risk of mycotoxins to humans. Qualitatively, the ADME processes seem to be quite similar between pigs and humans. In addition, similar metabolite and excretion patterns are observed, although some quantitative differences are noticed which are subject of this review. The high sensitivity of pigs towards mycotoxins and the similar kinetics are an advantage for the use of this animal species in the risk assessment of mycotoxins, and for the establishment of legal limits of mycotoxins. • Concerning toxicokinetics, the pig is a promising model in the assessment of the toxicological risk of mycotoxins to humans. • Qualitatively, the ADME processes seem to be quite similar between pigs and humans. • Similar metabolite and excretion patterns are observed, although some quantitative differences are noticed. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) and aflatoxin B1 (AFB1) are mycotoxins highly prevalent worldwide and can impair the performance and health of pigs. Preventive solutions such as the use of dietary mycotoxin deactivators (MD) can help in reducing the exposure to ZEN and AFB1 in animals. The aim of this study was to perform a toxicokinetic study in pigs focusing on the plasma concentration-time profiles of these 2 toxins, alone or in combination with a MD. The study was conducted on 12 healthy pigs (6 males/6 females) of 8 wk of age with an initial average body weight (BW) of 13.8 kg (ethical approval n° 2017/106). The acclimatization period was 1 wk (d 1-7). The pigs were randomly allocated into 2 mixed sex treatment groups of 6 pigs per group. On d 8, after fasting for 12 h, the pigs were given an intragastrical single bolus of ZEN and AFB1 (group 1: ZEN 0.5 mg/ kg BW and AFB1 0.1 mg/kg BW) or the same ZEN and AFB1 challenge with a MD (Unike Plus at 0.1 g/kg BW; group 2). Oral absorption and systemic exposure of ZEN and AFB1 were evaluated by comparing the plasma toxicokinetic parameters of the glucuronide conjugate of ZEN (ZEN-GlcA) and AFB1. For ZENGlcA, the area under the instrument response-time curve (AUC0-4h) for group 1 was 616.9 responses*h whereas it decreased to 379.2 response*h with the addition of MD. Therefore, MD reduced the oral bioavailability of ZEN by 38.5%. For AFB1, the area under the plasma concentration-time curve (AUC0-4h) for group 1 was 32.3 ng/mL*h whereas it decreased to 14.3 ng/ mL*h with the addition of MD. Therefore, the oral bioavailability of AFB1 with MD is 44.3% compared with group 1. This toxicokinetic model developed for swine has shown a great sensitivity that allows to quantity different levels of mycotoxin exposure. Unike Plus was able to reduce the systemic exposure to ZEN as well as AFB1. This preventive strategy is necessary to limit the appearance of clinical symptoms, and therefore, can help to control the development of infectious diseases and maintain the performance of animals. [ABSTRACT FROM AUTHOR]

To explore potential factors contributing to high fluoroquinolone resistance levels, it is essential to develop analytical methods capable of detecting residues and trace amounts of antibiotic use in broilers. The aim of the present study was to develop and in-house validate a sensitive UHPLC-MS/MS method capable of determining enrofloxacin (ENR) and flumequine (FLU) residues at slaughter age (day 45) when the animals were treated with these antimicrobials one day after hatching. Residue depletion of ENR and FLU in feathers was also assessed. Two experimental trials were performed, both consisting of 5 different treatment groups. In the first trial animals were treated with ENR and in the second one with FLU. The developed method was successfully validated and was found to be sensitive enough to detect residues of fluoroquinolones in the feathers up until slaughter age in all treatment groups. Average ENR concentration on day 45 was 10 ng g−1 feather after drinking water treatment, with all concentrations above the limit of quantification (LOQ) of 5 ng g−1 feather. For FLU average concentration on day 45 after drinking water administration was 4 ng g−1 feather, with an LOQ of 1 ng g−1 feather. Therefore, the method is suited for application to monitor fluoroquinolone use in broilers. [ABSTRACT FROM AUTHOR]

The use of juvenile conventional pigs as a preclinical animal model to perform pharmacokinetic (PK), pharmacodynamic (PD) and safety studies for the paediatric population is increasing. Repetitive oral administration of drugs to juvenile pigs is however challenging. A representative method which can be used from birth till adulthood is necessary. The current study presents the placement and use of a gastrostomy button in pigs with a weight ranging from 2.4 to 161 kg. The surgical placement was performed via a laparotomic procedure on, each time, 12 pigs (six male, six female) of 1 week, 4 weeks, 8 weeks and 6–7 months old. For every age category, eight pigs were part of a PK study with a non-steroidal anti-inflammatory drug (NSAID) and four pigs served as a control group. No severe complications were observed during surgery. The button remained functional for 10 days in 40 out of 48 pigs. No significant differences in body temperature or white blood cell count were observed during the trial. Three control pigs showed signs of inflammation indicating a NSAID might be warranted. Autopsy revealed minimal signs of major inflammation in the abdominal cavity or the stomach. A limited number of pigs showed mucosal inflammation, ulcers or abscesses in the stomach or around the fistula. These results indicate that the laparotomic placement of a gastrostomy button might be considered safe and easy in growing pigs to perform repetitive oral dosing preclinical studies. However, the method is not advised in pigs weighing more than 100 kg. [ABSTRACT FROM AUTHOR]

Background and Objective: Drug dosing should ideally be based on the drug concentrations at the target site, which, for most drugs, corresponds to the tissue. The exact influence of growth and development on drug tissue distribution is unclear. This systematic review compiles the current knowledge on the tissue distribution of systemically applied drugs in children, with the aim to identify priorities in tissue pharmacokinetic (PK) research in this population. Methods: A systematic literature search was performed in the MEDLINE and Embase databases. Results: Forty-two relevant articles were identified, of which 71% investigated antibiotics, while drug classes from the other studies were anticancer drugs, antifungals, anthelmintics, sedatives, thyreostatics, immunomodulators, antiarrhythmics, and exon skipping therapy. The majority of studies (83%) applied tissue biopsy as the sampling technique. Tonsil and/or adenoid tissue was most frequently examined (70% of all included patients). The majority of studies had a small sample size (median 9, range 1–93), did not include the youngest age categories (neonates and infants), and were of low reporting quality. Due to the heterogeneous data from different study compounds, dosing schedules, populations, and target tissues, the possibility for comparison of PK data between studies was limited. Conclusion: The influence of growth and development on drug tissue distribution continues to be a knowledge gap, due to the paucity of tissue PK data in children, especially in the younger age categories. Future research in this field should be encouraged as techniques to safely investigate drug tissue disposition in children are available. [ABSTRACT FROM AUTHOR]

To evaluate the cardiovascular effects, pharmacokinetic (PK) data and recovery characteristics of an alfaxalone constant rate infusion (CRI) of different duration in dogs at manufacturer's recommended dose rate. Experimental, prospective, randomized, crossover study. Six intact female Beagles. Following an intravenous alfaxalone bolus (3 mg kg–1), anaesthesia was maintained using an alfaxalone CRI at 0.15 mg kg–1 minute–1 for 90 (short CRI) or 180 minutes (long CRI). Venous blood samples were collected to determine the PK profile. Cardiovascular variables and recovery characteristics were evaluated. Recovery was scored on a scale ranging from 0, excellent to 4, bad. A mixed-model statistical approach was used to compare the cardiovascular parameters (global α = 0.05). An analysis of variance was performed to compare PK parameters and recovery times between treatments. No significant difference was noted between protocols for any PK parameter. Volume of distribution at steady state (935.74 ± 170.25 versus 1119.15 ± 190.65 mL kg–1), elimination half-life (12 ± 2 versus 13 ± 3 minutes), clearance from the central compartment (26.02 ± 4.41 versus 27.74 ± 5.65 mL kg–1 minute–1) and intercompartmental clearance (8.47 ± 4.06 versus 12.58 ± 7.03 mL kg–1 minute–1) were comparable for short CRI and long CRI. Cardiovascular variables remained within physiological limits. Mechanical ventilation was necessary (short CRI: n = 1, long CRI: n = 4). The manufacturer's recommended dose rate resulted in a light plane of anaesthesia. No significant differences in recovery times and scores were observed between treatments. The quality of recovery was scored as very poor with both protocols. PK data were similar between long and short infusions of alfaxalone at the manufacturer's recommended dose, with acceptable cardiovascular conditions. Nevertheless, both protocols resulted in a superficial plane of general anaesthesia with poor recovery characteristics. [ABSTRACT FROM AUTHOR]

Background: Asymptomatic malaria transmission has become a public health concern across malaria-endemic Africa including Ethiopia. Specifically, Plasmodium vivax is more efficient at transmitting earlier in the infection and at lower densities than Plasmodium falciparum. Consequently, a greater proportion of individuals infected with P. vivax can transmit without detectable gametocytaemia. Mass treatment of livestock with macrocyclic lactones (MLs), e.g., ivermectin and doramectin, was suggested as a complementary malaria vector tool because of their insecticidal effects. However, the effects of MLs on P. vivax in Anopheles arabiensis has not yet been fully explored. Hence, comparative in-vitro susceptibility and ex-vivo studies were conducted to evaluate the effects of ivermectin, doramectin and moxidectin sub-lethal concentrations on P. vivax oocyst development in An. arabiensis. Methods: The 7-day sub-lethal concentrations of 25% (LC25) and 5% (LC5) were determined from in-vitro susceptibility tests on female An. arabiensis in Hemotek® membrane feeding assay. Next, an ex-vivo study was conducted using P. vivax gametocytes infected patient's blood spiked with the LC25 and LC5 of the MLs. At 7-days post-feeding, each mosquito was dissected under a dissection stereo microscope, stained with 0.5% (w/v) mercurochrome solution, and examined for the presence of P. vivax oocysts. Statistical analysis was based on a generalized mixed model with binomially distributed error terms. Results: A 7-day lethal concentration of 25% (LC25, in ng/mL) of 7.1 (95% CI: [6.3;8.0]), 20.0 (95%CI:[17.8;22.5]) and 794.3 (95%CI:[716.4;1516.3]) were obtained for ivermectin, doramectin and moxidectin, respectively. Similarly, a lethal concentration of 5% (LC5, in ng/mL) of 0.6 (95% CI: [0.5;0.7]), 1.8 (95% CI:[1.6;2.0]) and 53.7 (95% CI:[ 48.4;102.5]) were obtained respectively for ivermectin, doramectin and moxidectin. The oocyst prevalence in treatment and control groups did not differ significantly (p > 0.05) from each other. Therefore, no direct effect of ML endectocides on P. vivax infection in An. arabiensis mosquitoes was observed at the sub-lethal concentration (LC25 and LC5). Conclusions: The effects of ivermectin and doramectin on malaria parasite is more likely via indirect effects, particularly by reducing the vectors lifespan and causing mortality before completing the parasite's sporogony cycle or reducing their vector capacity as it affects the locomotor activity of the mosquito. [ABSTRACT FROM AUTHOR]

The cross-contamination of non-medicated feed with residues of anti-microbials (AM) causes a public and animal health concern associated with the potential for selection and dissemination of resistance. To analyse the associated risks, a probabilistic model was built using @Risk® (Palisade Corporation®) to show the potential extent of the effect of cross-contaminated pig feed on resistance selection. The results of the model include estimations of the proportion of pigs per production stage with residues of doxycycline, chlortetracycline, sulfadiazine and trimethoprim in their intestinal contents, as a result of exposure to cross-contaminated feed with different carry-over levels, in Belgium. By using a semi-quantitative approach, these estimations were combined with experimental data on AM concentrations associated with potential for resistance-selection pressure. Based on this model, it is estimated that 7.76% (min = 1.67; max = 36.94) of sows, 4.23% (min = 1.01%; max = 18.78%) of piglets and 2.8% (min = 0.51%; max = 14.9%) of fatteners in Belgium have residues of doxycycline in their intestinal tract due to consumption of feed with at least 1% carry-over. These values were estimated to be almost triple for sulfadiazine, but substantially lower for chlortetracycline and trimethoprim. Doxycycline concentrations as low as 1 mg/L (corresponding to consumed feed with at least 1% carry-over) can select for resistant porcine commensal Escherichia coli in vitro and in vivo. Conclusions on this risk could not be drawn for other AM at this stage, due to the lack of data on concentrations associated with resistance development. However, since the possibility of resistance mechanisms (e.g. co-selection) occurring cannot be excluded, the results of this model highlight that the use of AM medicated feed should be minimised where possible. In case of medicated feed production, good practice should be followed thoroughly at all levels of production, distribution, storage and administration, with a special focus on the feed distributed to piglets and sows. [ABSTRACT FROM AUTHOR]

The Fusarium mycotoxin enniatin B (ENN B) is a so-called emerging mycotoxin frequently contaminating poultry feed. To investigate the impact of chronic ENN B exposure on animal health, broiler chickens were fed either a diet naturally contaminated with ENN B (2352 μg/kg) or a control diet (135 μg/kg) for 2, 7, 14, or 21 days. ENN B concentrations were determined in plasma and liver using a validated ultra-high performance liquid chromatography--tandem mass spectrometry UHPLC-MS/MS method. Liver was evaluated histologically, and the villus length and crypt depth of the duodenum, jejunum, and ileum were measured. Histopathology of the livers did not reveal major abnormalities. Feeding an ENN B-contaminated diet could possibly inhibit the proliferation of enterocytes in the duodenal crypts, but did not affect villus length, crypt depth, or villus length-crypt depth ratio of the jejunum and ileum. ENN B levels in plasma and liver were significantly higher in the ENN B-fed group and ranged between <25-264 pg/mL and <0.05-0.85 ng/g, respectively. ENN B carry-over rates from feed to liver tissue were 0.005-0.014% and 0.034-0.109% in the ENN B and control group, respectively. Carry-over rates were low and indicated a limited contribution of poultry tissue-derived products to the total dietary ENN B intake for humans. The above results support the opinion of the European Food Safety Authority stating that adverse health effects from ENN B in broiler chickens are unlikely. [ABSTRACT FROM AUTHOR]

Enniatins (ENNs) are hexadepsipeptidic mycotoxins produced by Fusarium species. They occur in mg/kg levels in grain from Northern climate areas. Major ENNs such as ENN B and B1 have shown considerable cytotoxicity in different in vitro test systems. To adequately assess exposure and in vivo toxicity the toxicokinetic properties need to be investigated. The present study describes the metabolism of ENN B1 both in vitro and in vivo in pigs, comparing metabolites found in vitro in experiments with liver microsomes from different pig strains to those found in the plasma of pigs after single oral or intravenous application of ENN B1. Metabolites of hepatic biotransformation were tentatively identified and characterised by high performance liquid chromatography coupled to ion trap and high-resolution mass spectrometry, as well as chemical derivatisations. Kinetic parameters of metabolite formation and elimination were determined. Metabolite formation was higher when ENN B1 was absorbed from the gut compared to intravenous administration indicating pre-systemic metabolism of ENN B1 after oral uptake. The in vitro approach resulted in the detection of ten ENN B1 metabolites, while six were detected in in vivo samples. The putative ENN B1 metabolites were products of hydroxylation, carbonylation, carboxylation and oxidative demethylation reactions. [ABSTRACT FROM AUTHOR]

Emerging Fusarium and Alternaria mycotoxins gain more and more interest due to their frequent contamination of food and feed, although in vivo toxicity and toxicokinetic data are limited. Whereas the Fusarium mycotoxins beauvericin, moniliformin and enniatins particularly contaminate grain and grain-based products, Alternaria mycotoxins are also detected in fruits, vegetables and wines. Although contamination levels are usually low (μg/kg range), higher contamination levels of enniatins and tenuazonic acid may occasionally occur. In vitro studies suggest genotoxic effects of enniatins A, A1 and B1, beauvericin, moniliformin, alternariol, alternariol monomethyl ether, altertoxins and stemphyltoxin-III. Furthermore, in vitro studies suggest immunomodulating effects of most emerging toxins and a reproductive health hazard of alternariol, beauvericin and enniatin B. More in vivo toxicity data on the individual and combined effects of these contaminants on reproductive and immune system in both humans and animals is needed to update the risk evaluation by the European Food Safety Authority. Taking into account new occurrence data for tenuazonic acid, the complete oral bioavailability, the low total body clearance in pigs and broiler chickens and the limited toxicity data, a health risk cannot be completely excluded. Besides, some less known Alternaria toxins, especially the genotoxic altertoxins and stemphyltoxin III, should be incorporated in risk evaluation as well. [ABSTRACT FROM AUTHOR]

Crossover animal trials were performed with intravenous and oral administration of deoxynivalenol-3-β- d-glucoside (DON3G) and deoxynivalenol (DON) to broiler chickens and pigs. Systemic plasma concentrations of DON, DON3G and de-epoxy-DON were quantified using liquid chromatography-tandem mass spectrometry. Liquid chromatography coupled to high-resolution mass spectrometry was used to unravel phase II metabolism of DON. Additionally for pigs, portal plasma was analysed to study presystemic hydrolysis and metabolism. Data were processed via tailor-made compartmental toxicokinetic models. The results in broiler chickens indicate that DON3G is not hydrolysed to DON in vivo. Furthermore, the absolute oral bioavailability of DON3G in broiler chickens was low (3.79 ± 2.68 %) and comparable to that of DON (5.56 ± 2.05 %). After PO DON3G administration to pigs, only DON was detected in plasma, indicating a complete presystemic hydrolysis of the absorbed fraction of DON3G. However, the absorbed fraction of DON3G, recovered as DON, was approximately 5 times lower than after PO DON administration, 16.1 ± 5.4 compared with 81.3 ± 17.4 %. Analysis of phase II metabolites revealed that biotransformation of DON and DON3G in pigs mainly consists of glucuronidation, whereas in chickens predominantly conjugation with sulphate occurred. The extent of phase II metabolism is notably higher for chickens than for pigs, which might explain the differences in sensitivity of these species to DON. Although in vitro studies demonstrate a decreased toxicity of DON3G compared with DON, the species-dependent toxicokinetic data and in vivo hydrolysis to DON illustrate the toxicological relevance and consequently the need for further research to establish a tolerable daily intake. [ABSTRACT FROM AUTHOR]

Seed-based pigeon diets could be expected to result in exposure of pigeons to mycotoxins such as deoxynivalenol (DON). Ingestion of low to moderate contamination levels of DON may impair intestinal health, immune function and/or pathogen fitness, resulting in altered host-pathogen interactions and thus different outcome of infections. Here we demonstrate that DON was one of the most frequently detected mycotoxins in seed-based racing pigeons feed, contaminating 5 out of 10 samples (range 177–1,466 μg/kg). Subsequently, a toxicokinetic analysis revealed a low absolute oral bioavailability (F) of DON in pigeons (30.4%), which is comparable to other avian species. Furthermore, semi-quantitative analysis using high-resolution mass spectrometry revealed that DON-3α-sulphate is the major metabolite of DON in pigeons after intravenous as well as oral administration. Following ingestion of DON contaminated feed, the intestinal epithelial cells are exposed to significant DON concentrations which eventually may affect intestinal translocation and colonization of bacteria. Feeding pigeons a DON contaminated diet resulted in an increased percentage of pigeons shedding Salmonella compared to birds fed control diet, 87 ± 17% versus 74 ± 13%, respectively. However, no impact of DON was observed on the Salmonella induced disease signs, organ lesions, faecal and organ Salmonella counts. The presented risk assessment indicates that pigeons are frequently exposed to mycotoxins such as DON, which can affect the outcome of a Salmonella infection. The increasing number of pigeons shedding Salmonella suggests that DON can promote the spread of the bacterium within pigeon populations. [ABSTRACT FROM AUTHOR]

Background: Cross-contamination of feed with low concentrations of antimicrobials can occur at production, transport and/or farm level. Concerns are rising about possible effects of this contaminated feed on resistance selection in the intestinal microbiota. Therefore, an experiment with pigs was set up, in which intestinal and fecal concentrations of chlortetracycline (CTC), doxycycline (DOX) and sulfadiazine-trimethoprim (SDZ-TRIM) were determined after administration of feed containing a 3 % carry-over level of these antimicrobials. Results: The poor oral bioavailability of tetracyclines resulted in rather high concentrations in cecal and colonic content and feces at steady-state conditions. A mean concentration of 10 mg/kg CTC and 4 mg/kg DOX in the feces was reached, which is higher than concentrations that were shown to cause resistance selection. On the other hand, lower mean levels of SDZ (0.7 mg/kg) and TRIM (< limit of detection of 0.016 mg/kg) were found in the feces, corresponding with the high oral bioavailability of SDZ and TRIM in pigs. Conclusions: The relation between the oral bioavailability and intestinal concentrations of the tested antimicrobials, may be of help in assessing the risks of cross-contaminated feed. However, future research is needed to confirm our results and to evaluate the effects of these detected concentrations on resistance selection in the intestinal microbiota of pigs. [ABSTRACT FROM AUTHOR]

Background: Despite the common use of oral group treatment in pig rearing, the magnitude of the factors influencing the homogeneity and stability of antimicrobial drugs in medicated feed and medicated drinking water are largely unknown, as well as the residual concentrations of the drugs after the end of the treatment. Results: This study presents a qualitative risk assessment to estimate the magnitude of the risks for reduced homogeneity and stability, and increased residual concentrations of antimicrobial drugs in medicated feed and drinking water on the farm. Risk assessment was done using a questionnaire and farm visits (n = 52), combined with a second questionnaire, and concentrations of amoxicillin and doxycycline measured in medicated feed and water samples, each collected on 10 farms. For medicated feed, the duration of storage in the silo did not show to influence the concentration levels in a consistent trend, while the treatment duration had a low to negligible effect. A moderate to high risk was found caused by human error when preparing the medicated feed on the farm. Purchased medicated feed greatly reduces the risk of human error and drugs remain stable during the duration of treatment, while the risk of residual concentrations after the end of the treatment was estimated to be low to moderate. The feed intake variability was identified as a moderate to high risk factor. For medicated drinking water, the type of dosing pump, age of pre-solution, and human errors during the preparation of the pre-solution present a moderate to high risk on homogeneity and stability. Precipitation of the active substance in the absence of a stirrer in a drinking water tank was shown to be a low to moderate risk factor for residues after treatment. Waterline length had a weak correlation with the concentrations of the antimicrobials, while a moderate to high influence was detected for the water intake by the pigs. Conclusions: A considerable variation in drug concentration in both medicated feed and medicated drinking water was detected depending on their preparation. Therefore, it is important to know which factors influence the homogeneity and stability, and the residual concentrations after treatment. [ABSTRACT FROM AUTHOR]

The most important source of Salmonella spp. infection in humans is by the consumption of contaminated poultry products. Due to the risk of resistance development and its transfer from animals to humans, the Belgian Royal Decree concerning the eradication of Salmonella (C-2007/22784) prohibits treatment of poultry with antimicrobials against zoonotic Salmonella spp. To uncover illicit use, an analytical method using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) for the determination of antimicrobial residues in poultry excreta was developed and validated for classes having an active spectrum against Salmonella spp. in poultry: β-lactams (amoxicillin and penicillin V), fluoroquinolones (enrofloxacin, difloxacin, and flumequine), polymyxins (colistin), sulfonamides in combination with trimethoprim (sulfachloropyridazine, sulfadiazine, and sulfaclozine), and tetracyclines (chlortetracycline and doxycycline). A generic and high-throughput sample preparation was developed. Extraction of samples was performed by ultrasonication using a combination of acetonitrile and McIlvaine buffer, followed by centrifugation and filtration prior to analysis. The method was validated according to Commission Decision 2002/657/EC for linearity, apparent recovery/trueness, repeatability, reproducibility, limit of quantification, limit of detection, specificity, matrix effect, and storage stability in matrix. To demonstrate the applicability of the method, an in vivo experiment was conducted. For each antimicrobial class, one registered drug was selected and administered in the drinking water to two laying hens. Excreta samples were collected every 12 h during and until 2 days after treatment and analyzed using the developed method. [ABSTRACT FROM AUTHOR]