Efficacy and safety testing of mycotoxin-detoxifying agents in broilers following the European Food Safety Authority guidelines.

Contamination of feeds with mycotoxins is a worldwide problem and mycotoxin-detoxifying agents are used to decrease their negative effect. The European Food Safety Authority recently stated guidelines and end-points for the efficacy testing of detoxifiers. Our study revealed that plasma concentrations of deoxynivalenol and deepoxy-deoxynivalenol were too low to assess efficacy of 2 commercially available mycotoxin-detoxifying agents against deoxynivalenol after 3 wk of continuous feeding of this mycotoxin at concentrations of 2.44±0.70 mg/kg of feed and 7.54±2.20 mg/kg of feed in broilers. This correlates with the poor absorption of deoxynivalenol in poultry. A safety study with 2 commercially available detoxifying agents and veterinary drugs showed innovative results with regard to the pharmacokinetics of 2 antibiotics after oral dosing in the drinking water. The plasma and kidney tissue concentrations of oxytetracycline were significantly higher in broilers receiving a biotransforming agent in the feed compared with control birds. For amoxicillin, the plasma concentrations were significantly higher for broilers receiving an adsorbing agent in comparison to birds receiving the biotransforming agent, but not to the control group. Mycotoxin-detoxifying agents can thus interact with the oral bioavailability of antibiotics depending on the antibiotic and detoxifying agent, with possible adverse effects on the health of animals and humans.

Lidocaine does not reduce pain perception during gel instillation sonography or subsequent office hysteroscopy: results of a randomized trial.

BACKGROUND: To evaluate if the addition of lidocaine to the gel used for gel infusion sonohysterography (GIS) reduces pain experienced during GIS or subsequent hysteroscopy. METHODS: A total of 142 consecutive patients were randomized using computer-generated random integers. In 79 patients, GIS was performed with a gel containing lidocaine (Instillagel®) and in 63 patients the gel did not contain lidocaine (Endosgel®). Immediately after GIS, 132 patients (94%) underwent office hysteroscopy. The women were asked to fill in a questionnaire including a 100-mm visual analogue scale (VAS) score after each examination. RESULTS: The mean age (SD) was 50.8 (12.1) years; 58.5% were premenopausal and 15.6% were nulliparous. The median (interquartile range (IR)) VAS score during GIS for all women was 6 (19.5): 8 (21) for the lidocaine group versus 5 (18.2) for those who received gel without lidocaine. The median (IR) VAS scores during hysteroscopy in the total group, the Instillagel group and the Endosgel group were 15.5 (43.2), 24 (35) and 9 (52), respectively. None of the differences were statistically significant. CONCLUSION: The addition of lidocaine to the gel used either for GIS or prior to office hysteroscopy does not reduce the procedure-related pain.

Routine gynecological ultrasound: look at the bladder and the ureters!

BACKGROUND: To illustrate the technical feasibility and diagnostic value of including the assessment of the bladder and the ureters during a standard transvaginal ultrasound examination. METHODS: We present four cases illustrating ureter and bladder pathology diagnosed by transvaginal ultrasound. RESULTS: In a first case, transvaginal ultrasonography in a woman with lower abdominal pain showed a calculus in the left distal ureter. The small echogenic lesion was easily detectable within the ureter lumen. A second patient, presenting with suprapubic pain, urgency and back pain, was also diagnosed with a ureteral calculus and additionally a hydroureter. The presence of a hydroureter is part of the differential diagnosis of any cystic structure in the pelvis. In a third case, an elderly woman, referred with uterine prolapse, transvaginal ultrasound examination showed a tubular anechoic structure lateral in the pelvis due to a hydroureter, illustrating the differential diagnosis with a hydrosalpinx. A fourth patient, presenting with hematuria, showed an irregular and highly vascularized mass in the bladder and was diagnosed with a transitional cell carcinoma. CONCLUSION: The bladder and pelvic part of the ureters are easily identifiable on transvaginal scan. Important pathology of the lower urinary tract and the distal ureters can be readily diagnosed by transvaginal ultrasound examination. The evaluation of the bladder and the ureters should therefore be part of the standard gynecological ultrasound investigation.

Gel infusion sonography in the evaluation of the uterine cavity.

OBJECTIVES: To compare gel infusion sonohysterography (GIS) with saline contrast sonohysterography (SCSH) with regard to technical feasibility and procedure-related pain experienced by patients. METHODS: This prospective observational cohort study included 551 consecutive patients with abnormal bleeding: SCSH was attempted in the first 402 women and GIS was attempted in the following 149. All procedures were performed by the same examiner, in the same clinical setting, using a 2-mm diameter catheter. After contrast sonohysterography, most patients underwent office hysteroscopy (n = 502) and endometrial sampling (n = 323). The women were asked to rate the pain experienced during each procedure using a 100-mm visual analog scale (VAS). Patients' characteristics, ultrasound findings, histological diagnosis, technical failures and procedure-related pain were compared between the two procedures. RESULTS: The mean +/- SD VAS score for contrast sonography, subsequent hysteroscopy and endometrial biopsy were 22.9 +/- 21.7, 38.8 +/- 26.6 and 50.0 +/- 26.3, respectively, in the SCSH subgroup vs. 16.5 +/- 21.5, 27.6 +/- 28 and 33.6 +/- 30.3, respectively, in the GIS subgroup (P = 0.0051, P = 0.0005 and P = 0.0003, respectively). The technical failure rate was 5% in the SCSH subgroup vs. 2% in the GIS subgroup (P = 0.1522). CONCLUSIONS: GIS and SCSH showed similar technical feasibility. The procedure-related pain reported by patients during contrast sonohysterography, as well as during subsequent hysteroscopy and endometrial sampling, was less in the GIS group.

Limitations of a chromogenic agar plate for the identifying bacteria isolated from equine endometritis samples.

BACKGROUND: The use of commercial chromogenic agar plates for the rapid, easy and correct identification of equine endometritis-causing bacteria has been proposed. Preliminary tests in our lab revealed undescribed limitations. Therefore, we tested the ability of the Brilliance UTI agar, a commercially available chromogenic agar, to accurately identify bacteria causing equine endometritis. OBJECTIVES: To 1) investigate whether bacteria present in the equine uterus are able to grow on this chromogenic agar plate, 2) determine whether these bacteria belong to the genera for which these agar plates were originally designed and 3) consider whether these bacterial genera can be correctly identified, based only on the colour appearance. STUDY DESIGN: In vitro experiments. METHODS: Macroscopic growth and colour appearance on the Brilliance UTI agar of 58 bacterial isolates, from previously collected equine uterine samples, were evaluated. Isolates were tentatively identified at the genus level using the manufacturer's guidelines and results were compared with MALDI-TOF MS as a "gold standard". RESULTS: The study revealed that 1) 77% (N = 45/58) of the bacterial isolates grew well on this chromogenic agar, 2) 83% of the investigated isolates (N = 48/58) belonged to one of the genera for which guidelines for identification were provided by the manufacturer and 3) only 50% of the isolates (N = 29/58) were correctly identified at the genus level, based only on colour appearance. MAIN LIMITATIONS: The current study uses purified bacterial isolates to inoculate the chromogenic agar plates, instead of fresh uterine samples. Bacteria were identified to the genus level using MALDI-TOF MS. CONCLUSION: This study shows that identification at the genus level based only on colour appearance, without additional tests or the simultaneous use of other media, is not reliable based on the existing identification guidelines.

Pain experienced during transvaginal ultrasound, saline contrast sonohysterography, hysteroscopy and office sampling: a comparative study.

OBJECTIVE: To evaluate and compare the pain experienced by women during transvaginal ultrasound, saline contrast sonohysterography (SCSH), diagnostic hysteroscopy and office sampling. METHODS: This was a descriptive study of 402 consecutive patients presenting at a 'one-stop' Bleeding Clinic between October 2004 and November 2006. Thirty-nine percent of the patients were postmenopausal. The patients underwent the following examinations transvaginally: first ultrasound with color Doppler, second SCSH, third diagnostic hysteroscopy and fourth endometrial biopsy. After completion of the examinations the patients were asked to complete a questionnaire including a visual analog scale (VAS) about their subjective appreciation of all four examinations. Two-hundred and ninety-three (72%) patients returned the questionnaire. RESULTS: The median (range) VAS scores for transvaginal ultrasound, SCSH, diagnostic hysteroscopy and endometrial sampling were 1.0 (0-8.1), 2.2 (0-10), 2.7 (0-10) and 5.1 (0-10), respectively (P < 0.0001). The patients' answers to the other questions about the pain experienced, including comparison with other minor procedures such as venous blood sampling, were all concordant with the VAS scores. CONCLUSIONS: Transvaginal ultrasound was the procedure best accepted, followed by SCSH, hysteroscopy and endometrial sampling. These results suggest that patients would prefer SCSH over hysteroscopy as an initial diagnostic approach in the evaluation of abnormal uterine bleeding.

Pharmacokinetics of eight anticoagulant rodenticides in mice after single oral administration.

The first aim of the study was to investigate the pharmacokinetics of eight anticoagulant rodenticides (brodifacoum, bromadiolone, chlorophacinone, coumatetralyl, difenacoum, difethialone, flocoumafen and warfarin) in plasma and liver of the mouse after single oral administration. Eight groups of mice dosed orally with a different anticoagulant rodenticide in a dose equal to one-half the lethal dose 50 (LD(50)), were killed at various times up to 21 days after administration. The eight anticoagulant rodenticides were assayed in plasma and liver by an LC-ESI-MS/MS method. Depending on the compound, the limit of quantification was set at 1 or 5 ng/mL in plasma. In liver, the limit of quantification was set at 250 ng/g for coumatetralyl and warfarin and at 100 ng/g for the other compounds. The elimination half-lives in plasma for first-generation rodenticides were shorter than those for second-generation rodenticides. Coumatetralyl, a first-generation product, had a plasma elimination half-life of 0.52 days. Brodifacoum, a second-generation product, showed a plasma elimination half-life of 91.7 days. The elimination half-lives in liver varied from 15.8 days for coumatetralyl to 307.4 days for brodifacoum. The second aim of the study was to illustrate the applicability of the developed method in a clinical case of a dog suspected of rodenticide poisoning.

Evaluation of the hygienogram scores and related data obtained after cleaning and disinfection of poultry houses in Flanders during the period 2007 to 2014.

Cleaning and disinfection (C&D) of poultry houses is an essential aspect in farm hygiene management. Adequate performance of the different steps of a C&D protocol and the use of suitable products are key to prevent and control zoonoses and animal diseases. Hygiene monitoring on total aerobic flora through sampling with agar contact plates at different locations of the poultry house results in a hygienogram score that is used to evaluate the proper execution of C&D.This study analyzed the hygienogram scores of 19,739 poultry flocks in Flanders after C&D. Data relating to the C&D protocol, i.e., year, season, husbandry system, production type, cleaning product, sampler, active components of the disinfectant, disinfection time, disinfection temperature, and disinfection responsible, were collected.The average hygienogram score decreased significantly over time, suggesting a general improvement between 2007 and 2014. Differences in scores were found among the husbandry systems, with the barn/aviary system having a significantly better hygienogram score compared to the floor house, furnished cage, and battery. Significantly better scores also were found when a cleaning product was used in the C&D protocol. Disinfection with a peracetic acid and hydrogen peroxide combination or formaldehyde gave the best scores. In addition, C&D protocols using ≥2 different disinfectants showed improved results compared to the use of one single disinfectant. Finally, disinfection applied by a specialist contractor resulted in a better score compared to disinfection by the farmer.In conclusion, analysis of the hygienogram scores and related data allowed identifying several factors, resulting in an improvement, which may reduce the total bacterial load in poultry stables and, consequently, the number of zoonotic and pathogenic micro-organisms.

Toxic effects of dietary exposure to T-2 toxin on intestinal and hepatic biotransformation enzymes and drug transporter systems in broiler chickens.

The effects of the mycotoxin T-2 on hepatic and intestinal drug-metabolizing enzymes (cytochrome P450) and drug transporter systems (MDR1 and MRP2) in poultry were investigated during this study. Broiler chickens received either uncontaminated feed, feed contaminated with 68µg/kg or 752µg/kg T-2 toxin. After 3weeks, the animals were euthanized and MDR1, MRP2, CYP1A4, CYP1A5 and CYP3A37 mRNA expression were analyzed using qRT-PCR. Along the entire length of the small intestine no significant differences were observed. In the liver, genes coding for CYP1A4, CYP1A5 and CYP3A37 were significantly down-regulated in the group exposed to 752µg/kg T-2. For CYP1A4, even a contamination level of 68µg/kg T-2 caused a significant decrease in mRNA expression. Expression of MDR1 was not significantly decreased in the liver. In contrast, hepatic MRP2 expression was significantly down-regulated after exposure to 752µg/kg T-2. Hepatic and intestinal microsomes were prepared to test the enzymatic activity of CYP3A. In the ileum and liver CYP3A activity was significantly increased in the group receiving 752µg/kg T-2 compared to the control group. The results of this study show that drug metabolizing enzymes and drug transporter mechanisms can be influenced due to prolonged exposure to relevant doses of T-2.

New bolus models for in vivo efficacy testing of mycotoxin-detoxifying agents in relation to EFSA guidelines, assessed using deoxynivalenol in broiler chickens.

In this study, three new models were developed for efficacy testing of mycotoxin-detoxifying agents in relation to recent European guidelines. In the first model, deoxynivalenol was given to broiler chickens as an intra-crop bolus together with a mycotoxin-detoxifying agent in order to study the plasma concentration-time profile of deoxynivalenol. In the second model, the same oral bolus was given, preceded by an oral bolus of mycotoxin-detoxifying agent, to make sure the detoxifying agent was present in the whole intestinal tract when the mycotoxin was administered. In the third model, the mycotoxin-detoxifying agent was mixed in the feed of broiler chickens, and after 1 week's feeding, deoxynivalenol was given as an oral bolus. In order to evaluate the efficacy of these agents, plasma concentration-time profiles were set up and the main toxicokinetic parameters were compared. Two commercially available mycotoxin-detoxifying agents were tested, but they were not able to lower the oral availability of deoxynivalenol. As a positive control, activated carbon was used. We showed that activated carbon significantly reduces the absorption and oral availability of deoxynivalenol in all three models. Therefore, it can be concluded that these models are able to demonstrate the efficacy of mycotoxin-detoxifying agents in relation to European Food Safety Authority guidelines.

Quantitative determination of T-2 toxin, HT-2 toxin, deoxynivalenol and deepoxy-deoxynivalenol in animal body fluids using LC-MS/MS detection.

A sensitive and specific method for the quantitative determination of deoxynivalenol (DON), deepoxy-deoxynivalenol (DOM-1), T-2 toxin (T-2) and HT-2 toxin (HT-2) in animal body fluids (plasma and bile) using liquid chromatography combined with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) is presented. The extraction of plasma consisted of a deproteinization step using methanol, followed by a clean-up using an Oasis HLB solid-phase extraction column. For bile analysis, an extraction using a methanol/water mixture (70/30, v/v), followed by a liquid-liquid extraction using ethyl acetate, was performed. Chromatographic separation was achieved on a reversed-phase Nucleosil (100-5 C18 G100 × 3.0 mm) column. For the analysis of DON and DOM-1, a mixture of 0.1% acetic acid in water and methanol was used as the mobile phase. T-2 and its metabolite HT-2 were separated using 5mM ammonium acetate in a mixture of water/methanol/acetic acid. The mass spectrometer was operated in the negative or positive ESI selected reaction monitoring mode for DON and T-2 analysis, respectively. Calibration graphs (1-250 ng mL(-1)) were prepared for all matrices and correlation and goodness-of-fit coefficients were between 0.9978-1.000 and 2.96-11.77%, respectively. Limits of quantification were between 1 and 2.5 ng mL(-1) for all compounds. Limits of detection ranged from 0.01 to 0.63 ng mL(-1). The results for the within-day precision and accuracy fell within the ranges specified. The method has been successfully used for the quantitative determination of DON, DOM-1, T-2 and HT-2 in plasma and the semi-quantitative determination of the same compounds in bile from broiler chickens and pigs, respectively.

Spark erosion of arteriosclerotic plaques.

For the recanalization of occluded arteries by means of plaque ablation an electrical technique, called spark erosion, has been developed. In vitro tests have shown that well-defined holes can be produced easily in fibrous, collagenous and lipid plaques, with minimal thermal side effects. As the method is not, by its nature, selective in vivo application will require an additional guidance technique to avoid arterial perforation. For this purpose, the potential use of tissue impedance as a sensing parameter has been tested. Main advantages of spark erosion are that it is a well controllable, low cost and highly effective tissue ablation technique.