Péter Laczay

Pharmacokinetics and bioavailability of doxycycline in fasted and nonfasted broiler chickens

The pharmacokinetics and the influence of food on the kinetic profile and bioavailability of doxycycline was studied after a single intravenous (i.v.) and oral dose of 10.0 mg/kg body weight in 7-week-old broiler chickens. Following i.v. administration the drug was rapidly distributed in the body with a distribution half-life of 0.21 +/- 0.01 h. The elimination half-life of 6.78 +/- 0.06 h was relatively long and resulted from both a low total body clearance of 0.139 +/- 0.007 L/h.kg and a large volume of distribution of 1.36 +/- 0.06 L/kg. After oral administration to fasted chickens, the absorption of doxycycline was quite fast and substantial as shown by the absorption half-life of 0.39 +/- 0.03 h, the maximal plasma concentration of 4.47 +/- 0.16 micrograms/mL and the time to reach the Cmax of 1.73 +/- 0.06 h. The distribution and the final elimination of the drug were slower than after i.v. administration. The absolute bioavailability was 73.4 +/- 2.5%. The presence of food in the intestinal tract reduced and extended the absorption (t1/2a = 1.23 +/- 0.21 h; Cmax = 3.07 +/- 0.23 micrograms/mL; tmax = 3.34 +/- 0.21 h). The absolute bioavailability was reduced to 61.1% +/- 4.4%.

Degradation of doxycycline in aged pig manure

Two types of studies were carried out to characterise the degradation rate of doxycycline in manure. In the laboratory experiment, pig manure containing doxycycline was stored in biochemical oxygen demand (BOD) bottles in a thermoregulated room at a temperature of 20 ± 3.5 °C to maintain the humid and anaerobic incubation conditions, and the relevant temperature. In the field study, the traditional manure composting process used by small farms and household plots was demonstrated, in which the manure heap containing doxycycline was exposed to the regular environmental conditions. In the laboratory experiment more than 30% and in the field study about 10% of the initial doxycycline amount could be detected in manure samples after 16 and 12 weeks of manure ageing period, respectively. Based on the results obtained from the studies performed under laboratory and field conditions, the half-life of doxycycline in manure was calculated to be 52.5 and 25.7 days, respectively.

Ecotoxicological assessment of doxycycline in soil

Doxycycline has been used in continually increasing quantities for mass treatment of food animals because of its greater bioavailability relative to older tetracyclines. The study presented in this paper was undertaken to investigate the degradation rate of the tetracycline derivative in manure-amended soil. In the present experiment, following composting, the doxycycline-contaminated manure was applied to agricultural land, and a field study was performed to investigate the degradation rate of doxycycline in soil. By the end of the 20-week sampling period, about 20 %, 33 % and 18 % of the initial doxycycline concentrations could be measured in soil samples taken at three different soil depths. The calculated half-life of doxycycline in the soil was 66.5, 76.3 and 59.4 days at depths of 0 cm, 25 cm and 50 cm, respectively. The potential effect of doxycycline on soil microbial activity was demonstrated by the nitrogen transformation test performed in compliance with the Organisation for Economic Co-operation and Development (OECD) Guideline No. 216. On day 28, the following nitrate concentrations of the control soil sample were found in the soil samples treated with different amounts of doxycycline: 76.9 %, 53.0 %, 65.6 %, 59.7 % and 77.1 %.

Determination of paromomycin residues in turkey tissues by liquid chromatography/mass spectrometry

A liquid chromatography-electrospray-mass spectrometry method has been developed and validated for the determination of paromomycin in turkey muscle, liver and kidney, using kanamycin as internal standard. The method consists of solid-phase extractions on mixed-mode columns. The chromatographic separation was carried out on a C(18) column using binary gradient elution containing acetonitrile and 5mM pentafluoropropionic-acid in water. The method was evaluated for specificity, linearity, recovery, accuracy, limit of detection, limit of quantification, intra- and inter-day repeatability, and stability. It was proven to be selective, linear, precise and accurate over the concentration range tested (0.5xMRL-2xMRL for each tissue) with correlation coefficients >0.990. The method was successfully used for the residue determination of PARO from edible tissues of turkeys.

Potentiation of ionophorous anticoccidials with dihydroquinolines: battery trials against Eimeria tenella in chickens.

Seven battery tests, employing a total of 1344 Hybro cockerels, were conducted in order to study the interaction between ionophorous anticoccidials and a new dihydroquinoline-type antioxidant known as duokvin. A significant, non-selective, toxic interaction was established, resulting in growth depression and improved anticoccidal efficacy against Eimeria tenella and E. mitis in these chickens. The duokvin itself showed no appreciable activity against the coccidia. The adverse effects of the interaction were eliminated, and the anticoccidial efficacy of the approved levels of ionophores was maintained, when the dietary levels of monovalent cation ionophorous monensin, salinomycin and narasin were reduced to approximately 12% in the presence of 120 p.p.m. duokvin. No adverse effects on the growth of chickens appeared in the combination with maduramicin, yet the enhancement of anticoccidial activity allowed an approx. 50% reduction of this ionophore as well.

Simultaneous determination of praziquantel, pyrantel embonate, febantel and its active metabolites, oxfendazole and fenbendazole, in dog plasma by liquid chromatography/mass spectrometry

A liquid chromatography-electrospray-mass spectrometry method (LC/MS) has been developed and validated for determination of praziquantel (PZQ), pyrantel (PYR), febantel (FBT), and the active metabolites fenbendazole (FEN) and oxfendazole (OXF), in dog plasma, using mebendazole as internal standard (IS). The method consists of solid-phase extractions on Strata-X polymeric cartridges. Chromatographic separation was carried out on a Phenomenex Gemini C6 -Phenyl column using binary gradient elution containing methanol and 50 mm ammonium-formate (pH 3). The method was linear (r(2)  ≥ 0.990) over concentration ranges of 3-250 ng/mL for PYR andFEB, 5-250 ng/mL for OXF and FEN, and 24-1000 ng/mL for PZQ. The mean precisions were 1.3-10.6% (within-run) and 2.5-9.1% (between-run), and mean accuracies were 90.7-109.4% (within-run) and 91.6-108.2% (between-run). The relative standard deviations (RSD) were <9.1%. The mean recoveries of five targeted compounds from dog plasma ranged from 77 to 94%.The new LC/MS method described herein was fully validated and successfully applied to the bioequivalence studies of different anthelmintic formulations such as tablets containing PZQ, PYR embonate and FBT in dogs after oral administration.

In vitro study on the effect of doxycycline on the microbial activity of soil determined by redox-potential measuring system

The potential effect of doxycycline on the microbial activity was investigated in three types of soil. Soil samples were spiked with doxycycline, incubated at 25°C and tested at 0, 2, 4 and 6 days after treatment. The microbiological activity of the soil was characterized by the viable count determined by plate pouring and by the time necessary to reach a defined rate of the redox-potential decrease termed as time to detection (TTD).The viable count of the samples was not changed during the storage. The TTD values, however exhibited a significant increase in the 0.2-1.6 mg/kg doxycycline concentration range compared to the untreated samples indicating concentration-dependent inhibitory effect on microbial activity. The potency of the effect was different in the 3 soil types. To describe the combined effect of the doxycycline concentration and time on the biological activity of one type of soil a mathematical model was constructed and applied.The change of microbial metabolic rate could be measured also without (detectable) change of microbial count when the traditional microbiological methods are not applicable. The applied new redox potential measurement-based method is a simple and useful procedure for the examination of microbial activity of soil and its potential inhibition by antibiotics.

RAPID DETECTION OF LISTERIA MONOCYTOGENES IN RAW MILK AND SOFT CHEESE BY A REDOX POTENTIAL MEASUREMENT BASED METHOD COMBINED WITH REAL-TIME PCR

The incidence of outbreaks of foodborne listeriosis has indicated the need for a reliable and rapid detection of the microbe in different foodstuffs. A method combining redox potential measurement and real-time polymerase chain reaction (PCR) was developed to detect Listeria monocytogenes in artificially contaminated raw milk and soft cheese. Food samples of 25 g or 25 ml were homogenised in 225 ml of Listeria Enrichment Broth (LEB) with Oxford supplement, and the redox potential measurement technique was applied. For Listeria species the measuring time was maximum 34 h. The absence of L. monocytogenes could reliably be proven by the redox potential measurement method, but Listeria innocua and Bacillus subtilis could not be differentiated from L. monocytogenes on the basis of the redox curves. The presence of L. monocytogenes had to be confirmed by real-time PCR. The combination of these two methods proved to detect < 10 cfu/g of L. monocytogenes in a cost- and time-effective manner. This method can potentially be used as an alternative to the standard nutrient method for the rapid detection of L. monocytogenes in food.

Rapid in-house detection method of Campylobacter spp. from food by redox potential monitoring combined with real-time PCR

The rapid detection of Campylobacter spp. is of utmost importance for the reduction of infections in humans by contaminated food products. The standard culturing method (ISO 10272-1:2006) involves a high time and labour demand. In this paper, we present a method that reduces the detection time of Campylobacter spp. to or below one third as compared to the ISO method, at a reduced cost per test. We used redox potential change of enrichment cultures (Bolton broth with Bolton selective supplement) for reliably selecting Campylobacter-contaminated raw milk and broiler meat samples. Identification of Campylobacter spp. in the contaminated samples was done by real-time PCR method. Culturing time to conclusive redox monitoring varied between 6 and 24 h for positive samples, depending on the contamination rate, in contrast to 136 h with the standard culturing process. However, now the Campylobacter-negative majority of food samples will not need to be tested by real-time PCR because redox potential monitoring can identify them in the selective enrichment phase. This method could be potentially used as a faster alternative to the current standard ISO 10272-1:2006, for nonregulatory monitoring purposes.