One-Pot facile synthesis of fluorescent copper nanoclusters for highly selective and sensitive detection of tetracycline.

Due to the excellent characteristics, fluorescent copper nanoclusters (Cu NCs) have aroused great interest in recent years. Herein, the simple prepared, environmentally friendly fluorescent Cu NCs were synthesized by using trypsin as the stabilizer and applied for the determination of tetracycline. Uniformly dispersed Try-Cu NCs were obtained with average size of 3.5 ± 0.3 nm and some excellent merits of good water solubility, UV light stability and salt stability. Emission peaks around 460.0 nm were visibly quenched by tetracycline based on static quenching mechanism and inner filter effect (IFE). Two excellent linear relationships were observed between ln(F0/F) and tetracycline concentrations in the range of 1-100 µM and 100-300 µM with limit of detection (LOD) of 0.084 µM. Meanwhile, this nanoprobe exhibited an apparent selectivity for tetracycline detection. Moreover, Try-Cu NCs were successfully employed to determine tetracycline in serum and milk samples after facile pretreatment with satisfactory recovery rates and credible standard deviation. The results suggested that this as-prepared Try-Cu NCs had excellent application prospects in the future.

A long lifetime ratiometrically luminescent tetracycline nanoprobe based on Ir(III) complex-doped and Eu3+-functionalized silicon nanoparticles.

Different from fluorescent dyes-doped or carbon materials-based ratiometric tetracycline nanoprobes, herein, a new Ir(III) complex-doped and europium(III) ion (Eu3+)-functionalized silicon nanoparticles (Ir(III)@SiNPs-Eu3+) with long luminescent lifetimes was firstly fabricated for selective detection of tetracycline (TC) in complex systems through time-resolved emission spectra (TRES) measurement. In the presence of TC, the red phosphorescence of Eu3+ is greatly enhanced by adsorption energy transfer emission (AETE) of TC, while the strong green luminescence of Ir(III)@SiNPs is quenched by the inner filtration effect (IFE) of TC. Based on these striking emission changes, Ir(III)@SiNPs-Eu3+ can sensitively detect TC in the linear range of 0.01-20 µM with a detection limit of 4.9 × 10-3 µM. Benefitting from the long lifetime of Ir(III)@SiNPs-Eu3+, the nanoprobe demonstrates excellent TC detection performance through TRES in high background system of 5 % human serum. Furthermore, the formed Ir(III)@SiNPs-Eu3+/TC complex can be used to sensitively recognize Hg2+ via a ratiometric luminescence mode. Notably, the cytotoxicity of Ir(III)@SiNPs-Eu3+ is very low and thus the sensitive monitoring the detection of Ir(III)@SiNPs-Eu3+ to TC and Hg2+ also works well in porcine renal cells, demonstrating high application potential in real samples.

Ratiometric detection of tetracycline based on gold nanocluster enhanced Eu3+ fluorescence.

Tetracycline (TC), a widely-used antibiotic to treat bacterial infections, combines with Eu3+ to form a stable EuTC complex that has a low fluorescence (FL) yield because of Eu3+ coordination with water molecules. We report a simple, label-free ratiometric fluorescent platform for sensitively and selectively sensing of TC, using L-histidine caped gold nanoclusters (His-AuNCs) as a FL indicator and an enhancer of Eu3+ FL. The His-AuNCs were prepared via chemical reduction of Au(III) by L-histidine, which was a reducer and a protecting agent. The His-AuNCs exhibited good photostability, outstanding stability toward high ionic strengths, storage stability, and favorable optical properties. In the absence of TC, the AuNCs-Eu3+ system displays strong FL emission at 475 nm (F475) from the His-AuNCs and weak FL at 620 nm from Eu3+ (F620) with excitation at 375 nm. TC quenches the His-AuNC FL and greatly enhances the Eu3+ emission. This is attributed to an enhancement of the EuTC complex fluorescence by the His-AuNCs. Thus, a ratiometric F620/F475 FL signal can be used for TC detection by simply mixing AuNCs and Eu3+. Under the optimized conditions, the linear range from 10 nM to 60 µM and a detection limit of 4 nM (S/N = 3) for TC were obtained. The application of the assay platform for the detection of TC in environmental and biological samples was demonstrated. The sensing platform has advantages of easy preparation, rapid response, high sensitivity, and good selectivity.

Development, validation and application of a novel HPLC-MS/MS method for the measurement of minocycline in human plasma and urine.

New treatments are urgently required to treat infections caused by multi-drug resistant Acinetobacter baumanni,. To address this need, a new formulation of Minocin®, (minocycline for injection) has been developed that allows for higher doses of minocycline to be administered. Phase 1 clinical trials were conducted in healthy volunteers to assess the safety and pharmacokinetics (PK) of this new formulation at higher doses. In order to generate PK data, novel, selective and simple HPLC-MS/MS based assays were developed and validated for the determination of minocycline (MC) in human plasma and urine. The respective working ranges were 0.05 to 30 mg/L and 0.1 to 30 mg/L. Removal of endogenous proteins with trichloroacetic acid was used as a simple means of extracting MC from the samples. An analogue, tetracycline was used as the internal standard (IS). Chromatographic separation, including that of MC from its 4-epimer (4-EMC), was achieved on a Waters XBridge BEH C18 column (50 x 4.6 mm ID, 5 µm) with gradient elution. The mobile phases comprised water containing 5 mM ammonium formate at a pH of 2.5, and methanol containing 5 mM ammonium formate. The internal standard (IS) was tetracycline, a structural analogue of minocycline. The methods were fully validated and met regulatory acceptance criteria for intra-run and inter-run accuracy and precision, carryover, dilution integrity and matrix effects. Mean extraction recoveries ranged between 64.3% and 84.6% for MC and 64.3% for the IS. There was no significant ion suppression or enhancement for MC or the IS. The validated assays were successfully applied to 1423 plasma and 689 urine samples from a Phase 1 clinical study. There was no evidence of instability, or significant interconversion between MC and 4-EMC, in stored clinical samples, spiked plasma and urine samples, or their extracts, under various test conditions.

Electrospun metal-organic framework/polyacrylonitrile composite nanofibrous mat as a microsorbent for the extraction of tetracycline residue in human blood plasma.

A new composite nanofiber of polyacrylonitrile doped with copper benzene-1,3,5-tricarboxylate metal-organic framework was fabricated by electrospinning and used as a microsorbent in the solid-phase extraction of trace tetracycline. The chemical structure of the adsorbent was studied by X-ray diffraction, scanning electron microscopy, Fourier transform infrared spectroscopy, Brunauer-Emmett-Teller surface area analysis, and Barrett-Joyner-Halenda pore size and volume analysis techniques. The significant parameters of the method including desorption solvent kind and volume, adsorbent mass, pH, and salt percentage were investigated. At the optimized conditions, the linear range was 8-1000 µg/L with a determination coefficient (R2 ) of 0.9954. The limits of detection and quantification were 2.40 and 8.00 µg/L, respectively. The inter- and intraday precisions were 4.7 and 3.4%, respectively. The developed extraction method was followed by high-performance liquid chromatography and applied for the determination of tetracycline in human blood plasma, and good relative recoveries (97.3-104.5%) were obtained.

Evaluation of tetracycline in milk following extra-label administration of topical tetracycline for digital dermatitis in dairy cattle.

Digital dermatitis (DD) is a painful infectious foot lesion commonly treated topically with extra-label tetracycline. Our objectives were to determine the concentrations of tetracycline in milk and plasma and to calculate a withdrawal interval following topical application at various doses. Another objective was to evaluate agreement between tests for measuring tetracycline in milk. A randomized block trial was conducted on 2 farms, where 50 cows with active DD lesions on 2 feet were allocated to 1 of 5 treatment groups (n = 10 cows per group). Treatment groups consisted of topical applications of tetracycline hydrochloride, in a paste or as a powdered form under a bandage, at 3 different dosing levels (2, 5, and 25 g) on each of the 2 affected feet. Following enrollment and treatment, samples were collected from milk, teat skin, and blood every 8 to 24 h for up to 7 d postdosing. Concentrations of tetracycline were measured by liquid chromatography-mass spectrometry and milk samples were further tested using the Charm ROSA TET test (Charm Sciences Inc., Lawrence, MA). Tetracycline was present in milk, plasma, and teat skin from all treatment groups. Tetracycline concentrations varied depending on time of sampling, method of application, and dosing level. At 8 h post-treatment, 11% of cows had tetracycline present in milk higher than 100 ng/mL (ppb) but none higher than 300 ng/mL. The 25-g treatment group had the longest estimated withdrawal interval, the highest observed concentrations (210-244 ng/mL) of tetracycline present in milk, and the longest observed consecutive period of tetracycline presence (from 8 to 72 h) among all treatment groups. Compared with liquid chromatography-mass spectrometry, the Charm test had a sensitivity of 77 and 100% for measuring tetracycline in milk at ≥30 and ≥100 ng/mL, respectively. Post-treatment samples of the teat skin were taken from 15 cows on 6 occasions, and every cow had tetracycline present in at least 1 of those 6 samples. This confirms an association between topical DD treatment with tetracycline and contamination of the teat. A total of 22% of blood samples had detectable tetracycline, and the majority (63%) occurred at 8 h post-treatment. At 100 ng/mL, the estimated cow-level milk withdrawal interval ranged from 0 to 70 h. At 300 ng/mL, the estimated cow-level withdrawal interval ranged from 0 to 34 h, and was 0 h at the bulk tank level. We recommend that conservative measures be adopted after extra-label use of topical tetracycline for DD treatment, including using a low dose and strategic post-treatment testing for tetracycline-class drugs in milk.

First four Oral Rabies Vaccination campaigns of the red foxes in Greece: Evaluating factors and assessment.

Following the last animal rabies outbreak in Greece in 2012, Oral Rabies Vaccination (ORV) campaigns of red foxes (Vulpes vulpes) were conducted in order to halt the spread of the disease, as widely and effectively have also been implemented in other countries. The present study aims to report the main outcomes following the first Greek ORV campaigns during autumn 2013, 2014, 2015 and spring 2016, to assess their effectiveness and to investigate factors potentially related to their success. Blood samples, mandible bones and teeth, derived by 452 foxes, were tested for rabies antibody titration, animal age determination and tetracycline (TTC) detection. The laboratory results obtained were statistically analyzed. High seroprevalence and TTC detection rates were obtained following the autumn campaigns studied, while these rates were significantly reduced following the spring campaign. The year or the season of the vaccination campaign, the estimated age group of the animal and the geographical Regional Unit (RU), where the animal was hunted, were identified as important factors. On the contrary, no significance could be ascertained for TTC detection based on exclusively previous uptake, use of filter paper, blood sample type and quality, as well as sex of animal. Based on the monitoring results achieved, the first ORV campaigns conducted in the country can be generally considered to be satisfactory. No positives cases were detected since May 2014. Seasonal, geographical parameters and factors related to fox ecology may interfere with monitoring results and should be always considered when planning future ORV programs.

Multianalyte Antibiotic Detection on an Electrochemical Microfluidic Platform.

The excessive use of antibiotics in human and veterinary medicine causes the emergence of multidrug resistant bacteria. In this context, the surveillance of many different antibiotics provokes a worldwide challenge. Hence, fast and versatile multianalyte single-use biosensors are of increasing interest for many fields such as medical analysis or environmental and food control. Here we present a microfluidic platform enabling the electrochemical readout of up to eight enzyme-linked assays (ELAs), simultaneously. To demonstrate the applicability of this platform for the surveillance and monitoring of antibiotics, we used highly sensitive biomolecular sensor systems for the simultaneous detection of two commonly employed antibiotic classes tetracycline and streptogramin. Thus, microfluidic channel networks are designed, comprising distinct numbers of immobilization sections with a very low volume of 680 nL each. These passively metered sections can be actuated separately for an individual assay procedure. The limits of detection (LOD) are determined, with high precision, to 6.33 and 9.22 ng mL-1 for tetracycline and pristinamycin, respectively. The employed channel material, dry film photoresist (DFR), allows an easy storage of preimmobilized assays with a shelf life of at least 3 months. Multianalyte measurements in a complex medium are demonstrated by the simultaneous detection of both antibiotics in spiked human plasma within a sample-to-result time of less than 15 min.

Fluorescence Probe Based on Hybrid Mesoporous Silica/Quantum Dot/Molecularly Imprinted Polymer for Detection of Tetracycline.

A newly designed fluorescence probe made from a hybrid quantum dot/mesoporous silica/molecularly imprinted polymer (QD/MS/MIP) was successfully created, and the probe was used for the detection of tetracycline (TC) in serum sample. QD/MS/MIP was characterized by transmission electron microscope, Fourier transform infrared spectroscopy, UV spectroscopy, X-ray powder diffraction, nitrogen adsorption-desorption experiment and fluorescence spectroscopy. Tetracycline, which is a type of broad-spectrum antibiotic, was selected as the template. The monomer and the template were combined by covalent bonds. After the template was removed to form a binding site, a hydrogen bonding interaction formed between the hole and the target molecule. Moreover, when rebinding TC, a new complex was produced between the amino group of QD/MS/MIP and the hydroxyl group of TC. After that, the energy of the QDs could transfer to the complex, which explains the fluorescence quenching phenomenon. The fluorescent intensity of QD/MS/MIP decreased in 10 min, and an excellent linearity from 50 to 1000 ng mL(-1) was correspondingly obtained. This composite material has a high selectivity with an imprinting factor of 6.71. In addition, the confirmed probe strategy was successfully applied to serum sample analyses, and the recoveries were 90.2%-97.2% with relative standard deviations of 2.2%-5.7%. This current work offers a novel and suitable method to synthesize QD/MS/MIP with a highly selective recognition ability. This composite material will be valuable for use in fluorescence probe applications.

Comparison of two fabricated aptasensors based on modified carbon paste/oleic acid and magnetic bar carbon paste/Fe3O4@oleic acid nanoparticle electrodes for tetracycline detection.

In this research, we have improved two aptasensors based on a modified carbon paste electrode (CPE) with oleic acid (OA), and a magnetic bar carbon paste electrode (MBCPE) with Fe3O4 magnetic nanoparticles and oleic acid (OA). After the immobilization process of anti-TET at the electrode surfaces, the aptasensors were named CPE/OA/anti-TET and MBCPE/Fe3O4NPs/OA/anti-TET respectively. In this paper, the detection of tetracycline is compared using CPE/OA/anti-TET and MBCPE/Fe3O4NPs/OA/anti-TET aptasensors. These modified electrodes were characterized by infrared spectroscopy (IR), scanning electron microscopy (SEM), electrochemical impedance spectroscopy (EIS), UV-vis spectroscopy, and voltammetric methods. The linear range and the detection limit for TET with the CPE/OA/anti-TET aptasensor were found to be 1.0×10(-12)-1.0×10(-7)M and 3.0×10(-13)M respectively by EIS method. The linear range and the detection limit for TET with the CPE/OA/anti-TET aptasensor were found to be 1.0×10(-10)-1.0×10(-7)M with a limit of detection of 2.9×10(-11)M using differential pulse voltammetry (DPV) technique. The MBCPE/Fe3O4NPs/OA/anti-TET aptasensor was used for determination of TET, and a liner range of 1.0×10(-14)-1.0×10(-6)M with a detection limit of 3.8×10(-15)M was obtained by EIS method. Also, the linear range and detection limit of 1.0×10(-12)-1.0×10(-6)M and 3.1×10(-13)M respectively, were obtained for MBCPE/Fe3O4NPs/OA/anti-TET aptasensor using DPV. The proposed aptasensors were applied for determination of tetracycline in some real samples such as drug, milk, honey and blood serum samples.

Efficient one-pot synthesis of hydrophilic and fluorescent molecularly imprinted polymer nanoparticles for direct drug quantification in real biological samples.

Efficient one-pot synthesis of hydrophilic and fluorescent molecularly imprinted polymer (MIP) nanoparticles and their application as optical chemosensor for direct drug quantification in real, undiluted biological samples are described. The general principle was demonstrated by preparing tetracycline (Tc, a broad-spectrum antibiotic)-imprinted fluorescent polymer nanoparticles bearing hydrophilic polymer brushes via poly(2-hydroxyethyl methacrylate) (PHEMA) macromolecular chain transfer agent-mediated reversible addition-fragmentation chain transfer (RAFT) precipitation polymerization in the presence of a fluorescent monomer. The introduction of hydrophilic PHEMA brushes and fluorescence labeling onto/into the MIP nanoparticles proved to not only significantly improve their surface hydrophilicity and lead to their obvious specific binding and high selectivity toward Tc in the undiluted bovine serum, but also impart them with strong fluorescent properties. In particular, significant fluorescence quenching was observed upon their binding with Tc in such complex biological milieu, which makes these Tc-MIP nanoparticles useful optical chemosensor with a detection limit of 0.26 µM. Furthermore, such advanced functional MIP nanoparticles-based chemosensor was also successfully utilized for the direct, sensitive, and accurate determination of Tc in another biological medium (i.e., the undiluted pig serum) with average recoveries ranging from 98% to 102%, even in the presence of several interfering drugs.

A novel colorimetric triple-helix molecular switch aptasensor for ultrasensitive detection of tetracycline.

Detection methods of antibiotic residues in blood serum and animal derived foods are of great interest. In this study a colorimetric aptasensor was designed for sensitive, selective and fast detection of tetracycline based on triple-helix molecular switch (THMS) and gold nanoparticles (AuNPs). As a biosensor, THMS shows distinct advantages including high stability, sensitivity and preserving the selectivity and affinity of the original aptamer. In the absence of tetracycline, THMS is stable, leading to the aggregation of AuNPs by salt and an obvious color change from red to blue. In the presence of tetracycline, aptamer binds to its target, signal transduction probe (STP) leaves the THMS and adsorbs on the surface of AuNPs. So the well-dispersed AuNPs remain stable against salt-induced aggregation with a red color. The presented aptasensor showed high selectivity toward tetracyclines with a limit of detection as low as 266 pM for tetracycline. The designed aptasensor was successfully applied to detect tetracycline in serum and milk.

Pharmacokinetics of metronidazole, tetracycline and bismuth in healthy volunteers after oral administration of compound tablets containing a combination of metronidazole, tetracycline hydrochloride and bismuth oxide.

BACKGROUND: To eradicate Helicobacter pylori in human pylorus and to heal duodenal ulcers, recently, a new formulation of combination tablets containing metronidazole 125 mg, tetracycline hydrochloride 125 mg and bismuth oxide 40 mg has been developed. OBJECTIVE: To investigate the pharmacokinetics of metronidazole, tetracycline and bismuth in healthy Chinese volunteers after oral administration of the test formulation. METHODS: A one-sequence, 3-period study was conducted in 12 Chinese healthy volunteers (6 male, 6 female). Volunteers each received single low dose (1 tablet) under fed condition in period 1, single high dose (3 tablets) under fasted condition in period 2, and single high dose (3 tablets) and multiple doses (3 tablets at once, 4 times daily for 7 consecutive days) under fed condition in period 3. Blood samples were collected and determined over 48 h in every period. RESULTS AND CONCLUSION: After single high dose administration under fed condition, the C max of metronidazole, tetracycline and bismuth were 6.833 ± 0.742 µg/mL, 0.8513 ± 0.1253 µg/mL and 3.32 ± 1.89 ng/mL, respectively. The C max and AUC 0-48 of metronidazole increased in proportion to the doses within the tested dose range, but tetracycline and bismuth did not. Food caused 10% and 80% decrease of the C max for metronidazole and bismuth, respectively, but did not affect tetracycline. No gender effect was found on the pharmacokinetics of the 3 ingredients. In the steady state, the C av of metronidazole, tetracycline and bismuth were 20.75 ± 3.52 µg/mL, 1.900 ± 0.243 µg/mL and 5.61 ± 1.34 ng/mL, respectively.

Novel and remarkable enhanced-fluorescence system based on gold nanoclusters for detection of tetracycline.

Tetracycline and Eu(3+), while coexisting, usually appear as a complex by chelating. This complex shows low fluorescence intensity, leading to its limitation of analytical goals. Gold nanoclusters (AuNCs), emerging as novel nano-material, are attracting increasing attentions in multiple fields. Herein, gold nanoclusters first function as a fluorescence-enhanced reagent rather than a conventional fluorescent-probe, and a dramatic enhanced-fluorescence system was built based on Eu(3+)-Tetracycline complex (EuTC) by introducing gold nanoclusters. Simultaneously, three types of gold nanoclusters were employed for exploring various conditions likely affecting the system, which demonstrate that no other gold nanoclusters than DNA-templated gold nanoclusters enormously caused fluorescence-enhancement of EuTC. Moreover, this enhanced-fluorescence system permitted available detection of tetracycline (TC) in a linear range of 0.01-5 µM, with a detection limit of 4 nM at a signal-to-noise ratio of 3. Significantly, the practicality of this method for detection of TC in human urine and milk samples was validated, demonstrating its advantages of simplicity, sensitivity and low cost. Interestingly, this system described here is probably promising for kinds of applications based on its dramatically enhanced-fluorescence.

Tetracycline residues in porcine stomach after administration via drinking water on a swine farm.

Tetracycline is a broad-spectrum antibiotic used to treat infections in swine. The maximum residue levels of tetracycline in pork stomach tissue in Russia, Europe, and the United States are 10, 200, and 2,000 ppb, respectively. This difference in accepted safety levels may be the reason why stomach tissues that the United States exports continue to be residue violators in overseas markets. In this study, 30 pigs at two different stages of production (weanling and finisher) were treated with tetracycline at 22 mg/kg of body weight per day for a total of 5 days via a water medicator. Blood samples were collected at 0, 72, 78, 96, and 102 h after the start of medication. The medication was stopped at 120 h, and blood samples were again collected at 126, 144, 168, 192, and 216 h after exposure. Five animals were slaughtered for stomach tissue 0, 24, 48, 96, and 192 h after the drug was flushed from the water line. All blood and tissue samples were analyzed by high-performance liquid chromatography-UV methods. The tetracycline levels in plasma were below the level of detection after the U.S.-labeled withdrawal time of 4 days. The stomach tissue residues averaged 671.72, 330.31, 297.77, 136.36, and 268.08 ppb on withdrawal days 0, 1, 2, 4, and 8, respectively. Using the U.S. Food and Drug Administration tolerance limit method and a population-based pharmacokinetic model with Monte Carlo simulation, a withdrawal interval was estimated. This study demonstrated that tetracycline residues are still detectable in the stomach tissues after the established United States withdrawal time of 4 days. These residue levels may explain why stomach tissues tested in Russia and Europe show positive residues for tetracycline, even though the meat may pass inspection here in the United States prior to export.

Aptamer based extraction followed by electrospray ionization-ion mobility spectrometry for analysis of tetracycline in biological fluids.

An extraction method based on aptamer sorbent followed by electrospray ionization-ion mobility spectrometry (ESI-IMS) has been developed for the analysis of tetracycline in human urine and plasma samples. The effect of extraction parameters on the extraction efficiency including washing (solvent type and volume) and elution (solvent type, volume and flow rate) were investigated. Under the optimized conditions, the linear dynamic ranges for tetracycline in urine and plasma samples were found to be 0.05-5.00 µg/mL and 0.10-5.00 µg/mL with detection limits of 0.019 and 0.037 µg/mL, respectively. The extraction efficiency was 86.5% for urine and it was 82.8% for plasma samples. The relative standard deviation was 5.9% and 6.3% for six replicate measurements of tetracycline at 1 and 2 µg/mL in urine and plasma samples, respectively.

Evaluation of factors important in modeling plasma concentrations of tetracycline hydrochloride administered in water in swine.

OBJECTIVE: To model the plasma tetracycline concentrations in swine (Sus scrofa domestica) treated with medication administered in water and determine the factors that contribute to the most accurate predictions of measured plasma drug concentrations. SAMPLE: Plasma tetracycline concentrations measured in blood samples from 3 populations of swine. PROCEDURES: Data from previous studies provided plasma tetracycline concentrations that were measured in blood samples collected from 1 swine population at 0, 4, 8, 12, 24, 32, 48, 56, 72, 80, 96, and 104 hours and from 2 swine populations at 0, 12, 24, 48, and 72 hours hours during administration of tetracycline hydrochloride dissolved in water. A 1-compartment pharmacostatistical model was used to analyze 5 potential covariate schemes and determine factors most important in predicting the plasma concentrations of tetracycline in swine. RESULTS: 2 models most accurately predicted the tetracycline plasma concentrations in the 3 populations of swine. Factors of importance were body weight or age of pig, ambient temperature, concentration of tetracycline in water, and water use per unit of time. CONCLUSIONS AND CLINICAL RELEVANCE: The factors found to be of importance, combined with knowledge of the individual pharmacokinetic and chemical properties of medications currently approved for administration in water, may be useful in more prudent administration of approved medications administered to swine. Factors found to be important in pharmacostatistical models may allow prediction of plasma concentrations of tetracycline or other commonly used medications administered in water. The ability to predict in vivo concentrations of medication in a population of food animals can be combined with bacterial minimum inhibitory concentrations to decrease the risk of developing antimicrobial resistance.

Pharmacokinetics of tetracycline after single-dose oral administration in the American alligator (Alligator mississippiensis).

The major objective of the study was to assess the pharmacokinetics of tetracycline administered orally to fasted and nonfasted American alligators (Alligator mississippiensis) at 50 mg/kg. Plasma levels of tetracycline were determined using high-performance liquid chromatography with ultraviolet detection. The concentration versus time curve was analyzed using a compartmental modeling technique. A one-compartment model with first-order absorption and elimination, as well as a lag time to absorption, best described the data. The area under the curve and mean residence time values differed significantly between the fasted and nonfasted groups. Based on the results of this study, tetracycline suspension administered once orally at 50 mg/kg to American alligators is not expected to reach plasma concentrations above the breakpoint minimum inhibitory concentration of 4 microg/ml for susceptible organisms.

Raccoon dog rabies surveillance and post-vaccination monitoring in Lithuania 2006 to 2010.

BACKGROUND: Oral rabies vaccination (ORV) in rabies infected regions should target the primary rabies vector species, which in Lithuania includes raccoon dogs as well as red foxes. Specific investigations on ORV in raccoon dogs are needed e.g. evaluation of vaccine effectiveness under field conditions. The objective of the current study was to investigate the efficacy of the ORV programme 2006-2010 in Lithuania by examining the number of rabies cases and estimating the prevalences of a tetracycline biomarker (TTC) and rabies virus antibodies in raccoon dogs. METHODS: From 2006 to 2010, 12.5 million rabies vaccine-baits were distributed by aircraft. Baiting occurred twice per year (spring and autumn), targeting raccoon dogs and red foxes in a 63,000 km2 area of Lithuania. The mandibles of raccoon dogs found dead or killed in the vaccination area were analyzed by fluorescence microscopy for the presence of the TTC. Rabies virus sera neutralizing anti-glycoprotein antibody titres were determined using an indirect ELISA method and seroconversion (> 0.5 EU/ml) rates were estimated. RESULTS: During the study period, 51.5% of raccoon dog mandibles were positive for TTC. 1688 of 3260 tested adults and 69 of 175 tested cubs were TTC positive. Forty-seven percent of raccoon dog serum samples were positive for rabies virus antibodies. 302 of 621 investigated adults and 33 of 95 investigated cubs were seropositive. In the same time 302 of 684 and 43 of 124 tested samples were TTC and ELISA positive in spring; whereas 1455 of 2751 and 292 of 592 tested samples were TTC and ELISA positive in autumn. There was a positive correlation between the number of TTC and antibody positive animals for both adult and cub groups. CONCLUSIONS: ORV was effective in reducing the prevalence of rabies in the raccoon dog population in Lithuania. The prevalence of rabies cases in raccoon dogs in Lithuania decreased from 60.7% in 2006-2007 to 6.5% in 2009-2010.

Farnesol as antibiotics adjuvant in Staphylococcus epidermidis control in vitro.

INTRODUCTION: Farnesol is a sesquiterpenoid that has been described as impairing bacterial growth. Therefore, the goal of this study was to compare the in vitro postantimicrobial effect (PAE) of farnesol against Staphylococcus epidermidis with the corresponding values of most common practice antibiotics and also to evaluate the combined effect of farnesol with these antibiotics against planktonic and biofilm cells. METHODS: After exposure of S epidermidis cells to farnesol and antibiotics at minimum inhibitory concentration for 1 hour, the cells were regrown in medium without any antimicrobial agent. Cellular viability was assessed by colony-forming units, every hour for 12 hours, and then, the PAE was determined. The combined effect of farnesol (0, 30, 100 and 300 µM) with vancomycin, tetracycline and rifampicin was also evaluated, by using these antibiotics at peak serum concentration. RESULTS: When PAE is concerned, it was found that cells grown in 100 µM of farnesol behaved similarly to cells that had never been in contact with farnesol, whereas a clear difference was obtained with cells exposed to 300 µM of farnesol, displaying a longer PAE. Farnesol showed a combined effect with the tested antibiotics against planktonic cells, although this was not so evident against biofilm cells. CONCLUSIONS: Despite the reduced efficacy against biofilm cells, farnesol seems to be a potential adjuvant therapeutic agent to antibiotics against S epidermidis planktonic cells. Moreover, its long PAE makes farnesol a potential candidate in the prevention of biofilm formation because it showed to be very effective against planktonic cells alone as well.