Antibiotics In Food Research Articles

Antibiotic residue monitoring results for pork, chicken, and beef samples in Vietnam in 2012-2013.

A monitoring plan of residual antibiotics in food of animal origin was conducted in Vietnam from 2012 to 2013. Meat samples were collected from slaughterhouses and retail stores in Ho Chi Minh City and Nha Trang. A total of 28 antibiotics were analyzed using a LC-MS/MS screening method. Sulfonamides, fluoroquinolones, and tilmicosin were detected in some of the samples. Sulfaclozine and fluoroquinolones were mainly detected in chicken samples, and sulfamethazine was mainly detected in pork samples. High levels of sulfonamide residues, ranging between 2500 and 2700 μg/kg sulfaclozine and between 1300 and 3600 μg/kg sulfamethazine, were present in two chicken and three pork samples, respectively. Tilmicosin was detected at ranges of 150-450 μg/kg in 10 chicken samples. Positive percentages were 17.3, 8.8, and 7.4% for chicken, pork, and beef, respectively, for an average of 11.9%. The results suggest an appropriate withdrawal period after drug administration had not been observed in some livestock.

A novel antibacterial tripeptide from Chinese leek seeds

Antibacterial peptides have been one of the most prospects as a new member of antibiotics in food and/or medicine industry. In this thesis, a novel antibacterial peptide, designated as CLP-1, was purified from Chinese leek seeds by a gel filtration chromatography and reversed-phase high-performance liquid chromatography. CLP-1 was composed of three amino acids, and the sequence was identified as Ser-Asn-Ala (SNA) using LC–MS/MS, which was a novel short antibacterial peptide from Chinese leek seeds. SNA showed moderate antibacterial activity against the growth of Gram-positive and Gram-negative bacteria, and the minimal inhibition concentration values of SNA against Escherichia coli, Staphylococcus aureus, Salmonella, and Bacillus subtilis were 4.31, 2.24, 4.31, and 2.24 mM, respectively. Moreover, SNA exerted slighter hemolytic activity at concentration of 3.44 mM.

Risk of allergic reaction and sensitization to antibiotics in foods

Risk of allergic reaction and sensitization to antibiotics in foods

Development of a direct ELISA based on carboxy-terminal of penicillin-binding protein BlaR for the detection of β-lactam antibiotics in foods

β-Lactam antibiotics, including penicillins and cephalosporins, are commonly used in veterinary medicine. Illegal use and abuse of β-lactams could cause allergy and selected bacterial resistance. BlaR-CTD, the carboxy-terminal of penicillin-recognizing protein BlaR from Bacillus licheniformis ATCC 14580, was utilized in this study to develop a receptor-based ELISA for detection and determination of β-lactam antibiotics in milk, beef, and chicken. This assay was based on directly competitive inhibition of binding of horseradish peroxidase-labeled ampicillin to the immobilized BlaR-CTD by β-lactams. The assay was developed as screening test with the option as semiquantitative assay, when the identity of a single type of residual β-lactam was known. The IC50 values of 15 β-lactam antibiotics, including benzylpenicillin, ampicillin, amoxicillin, dicloxacillin, oxacillin, nafcillin, cefapirin, cefoperazone, cefalotin, cefazolin, cefquinome, ceftriaxone, cefotaxime, cefalexin, ceftiofur and its metabolite desfuroylceftiofur were evaluated and ranged from 0.18 to 170.81 μg L(-1). Simple sample extraction method was carried out with only phosphate-buffered saline, and the recoveries of selected β-lactam antibiotics in milk, beef, and chicken were in the range of 53.27 to 128.29 %, most ranging from 60 to 120 %. The inter-assay variability was below 30 %. Limits of detection in milk, beef, and chicken muscles with cefquinome matrix calibration were 2.10, 30.68, and 31.13 μg kg(-1), respectively. This study firstly established a rapid, simple, and accurate method for simultaneous detection of 15 β-lactams in edible tissues, among which 11 β-lactams controlled by European Union could be detected below maximum residue limits.

Highly sensitive and selective colorimetric sensing of antibiotics in milk

Antibiotics residues in foods are very harmful to human beings. Determination of antibiotics residues relies largely on the availability of adequate analytical techniques. Currently, there is an urgent need for on site and real time detection of antibiotics in food. In this work, a novel one step synthesis of gold nanoparticles (AuNPs) was proposed using pyrocatechol violet (PCV) as a reducer agent. Highly sensitive and selective colorimetric detection of four antibiotics kanamycin mono sulfate (KA), neomycin sulfate (NE), streptomycin sulfate (ST) and bleomycin sulfate (BL) was realized during the formation of AuNPs. PCV has OH groups and these antibiotics have OH, NH2, NH groups, so there may be some special hydrogen-bonding interactions between PCV and these antibiotics. Therefore, the presence of KA, NE, ST and BL would influence the synthesis of AuNPs, then the color and state of AuNPs would change, which could be observed with the naked eye or a UV–vis spectrophotometer. Results showed that A670 was linear with the logarithm of KA concentration in the range from 1.0×10−8 to 5.0×10−7M and 5.0×10−7 to 5.5×10−5M. The detection limit of KA was 1.0×10−9M (S/N=3). The coexisting substances including 1.0×10−5M phenylalanine, alanine, glycerol, glucose, Mg2+, Ca2+, Na+, K+, CO32−, SO42−, NO3−, Cl− and Br− did not affect the determination of 1.0×10−7M antibiotics. In particular, the proposed method could be applied successfully to the detection of antibiotics in the pretreated liquid milk products.

Zwischen Verrheißung und Bedrohung. Antibiotika in Nahrungsmitteln in Westdeutschland 1950–1980

Produktion und Verbrauch von Nahrungsmitteln sind traditionell mit weit reichenden Angsten verknupft. Bezogen diese sich in vormodernen Gesellschaften vorrangig auf eine ausreichende Versorgung, sind sie in der modernen Industriegesellschaft eher auf die Qualitat der Nahrungsmittel gerichtet. In diesem Zusammenhang spielten in der Nachkriegszeit Antibiotika eine zentrale Rolle. Der Aufsatz stellt die Einfuhrung der Antibiotika in die westdeutsche Veterinarmedizin, ihre fruhe Diskussion als Konservierungsmittel und ihre wachsende Rolle als Wachstumsforderer dar. Vor dem Hintergrund einer zunehmend industrialisierten, auf Maximierung ausgerichteten Produktionspolitik werden die Rolle der Veterinare, der Forschung und der wissenschaftlichen Experten sowie die Probleme der Regulierung des Antibiotikaeinsatzes in der Landwirtschaft diskutiert, die in die Entstehung eines unubersichtlichen grauen Marktes fur Antibiotika und die Verlagerung ihres Einsatzes in die Therapie mundete. Besondere Aufmerksamkeit wird der Frage zugewandt, welche Rolle das Vertrauen und die Angste der Verbraucher in diesem Prozess spielten.

An Ex Vivo Model for Suture-Less Amniotic Membrane Transplantation with a Chemically Defined Bioadhesive

Purpose: Chemically defined bioadhesives (CDB) are “two-component reactive adhesives” made from antibiotic food additives. They contain no animal-derived ingredients and are characterized by a high level of self-degradability, low toxicity, and strong bonding properties. Amniotic membrane (AM) transplantation is used to repair severe ocular surface disease in humans and horses. AM requires meticulous suturing to the injured cornea. The purpose of this study is to determine whether CDB can facilitate the suture-less placement and adherence of equine AM to porcine cadaver cornea.Design: Prospective study.Materials and Methods: To determine whether there are differences in ex vivo adhesion force between equine AM bonded with CDB to porcine cadaver cornea and equine AM sutured to porcine cadaver cornea. The following groups were tested (n = 5 per group): (1) CDB group: trephined (8 mm in diameter) cornea was bonded with CDB to 8-mm-sized discs of AM. (2) Suture group: the 8-mm trephined cornea was sutured with 8–0 absorbable suture to the AM. (3) Control group 1: two sheets of nitrocellulose paper were bonded with CDB. (4) Control group 2: two sheets of nitrocellulose paper were attached with suture and tested. Each sample was anchored to a tensiometer materials testing machine, and the strength of adhesion measured and expressed in units of kilogram-force (kgf).Results: Mean ± SD strength of adhesion force found no statistically significant difference between CDB group (0.091 ± 0.054 kgf; 15 min of CDB adhesion time) and suture groups (0.095 ± 0.037 kgf), whereas there was significant difference (P < 0.05) between control group 1 (0.679 ± 0.048 kgf) and control group 2 (0.585 ± 0.092 kgf). These results revealed that the 15 min of CDB adhesive time had adhesive properties as strong as suture group.Conclusions and Clinical Relevance: CDB provides a satisfactory level of adherence to ex vivo suture-less attachment of equine amniotic membrane transplantation to the porcine cornea.

Optimisation and validation of a quantitative and confirmatory method for residues of macrolide antibiotics and lincomycin in kidney by liquid chromatography coupled to mass spectrometry

A solid phase extraction followed by a liquid chromatography (LC)‐tandem mass spectrometry (MS/MS) detection method for the confirmatory analysis of lincomycin (LIN), clindamycin (CLI), tilmicosin (TIM), erythromycin (ERI) and tylosin (TYL) residues in kidney were optimised and validated for monitoring and controlling the use of these antibiotics in food producing‐animals. The method optimisation was carried out by testing changes in the extraction buffer pH and in the ammonium/acetonitrile concentrations on SPE eluent solutions. The optimised extraction procedure involved the extraction of the analytes with a pH 8 phosphate buffer, clean‐up on a reversed‐phase mixed‐cation exchange cartridge, followed by the elution of the analytes in a 98:2 acetonitrile/ammonia solution, concentration in air flow and re‐dissolved with an 1:1 methanol/water solution. The analytes were detected in an LC‐MS/MS system in electrospray positive ionisation mode. The validation was performed according to the European Commission Decision 2002/657/EC. Linearity was established for all analytes using the method of least weighted squares and CCα values ranged from 5.3% to 21.1% higher than the minimum residue limit (MRL) values. The addition levels varied from 0.5 to 1.50 MRL for all analytes, with recoveries exceeding 92.5%. The relative standard deviations (RSD%) in terms of repeatability (n = 54) and reproducibility (n = 108) for all analytes were less than 21.6% and 21.4%, respectively. The uncertainties were calculated by simplified methods using the calibration curve uncertainty and the intermediate precision to obtain the combined measurement uncertainty. The results of the validation process demonstrated that this method is suitable for the quantification and confirmation of antibiotic residues for the Brazilian Residue and Contaminant Control Plan (PNCR).

Impact of enrofloxacin on the human intestinal microbiota revealed by comparative molecular analysis

The indigenous human intestinal microbiota could be disrupted by residues of antibiotics in foods as well as therapeutically administered antibiotics to humans. These disruptions may lead to adverse health outcomes. To observe the possible impact of residues of antibiotics at concentrations below therapeutic levels on human intestinal microbiota, we performed studies using in vitro cultures of fecal suspensions from three individuals with 10 different concentrations (0, 0.1, 0.5, 1, 5, 10, 15, 25, 50 and 150 μg/ml) of the fluoroquinolone, enrofloxacin. The bacterial communities of the control and enrofloxacin dosed fecal samples were analyzed by denaturing gradient gel electrophoresis (DGGE) and pyrosequencing. In addition, changes of functional gene expression were analyzed by a pyrosequencing-based random whole-community mRNA sequencing method. Although each individual had a unique microbial composition, the communities of all individuals were affected by enrofloxacin. The proportions of two phyla, namely, Bacteroidetes and Proteobacteria, were significantly reduced with increasing concentrations of enrofloxacin exposure, while the proportion of Firmicutes increased. Principal Coordinate Analysis (PCoA) using the Fast UniFrac indicated that the community structures of intestinal microbiota were shifted by enrofloxacin. Most of the mRNA transcripts and the anti-microbial drug resistance genes increased with increasing concentrations of enrofloxacin. 16S rRNA gene pyrosequencing of control and enrofloxacin treated fecal suspensions provided valuable information of affected bacterial taxa down to the species level, and the community transcriptomic analyses using mRNA revealed the functional gene expression responses of the changed bacterial communities by enrofloxacin.

Dysprosium-sensitized chemiluminescence reactions: Their mechanism and application to the determination of synthetic quinolone antibiotics

Dysprosium-sensitized chemiluminescence (CL) reactions have been suggested for the determination of enoxacin (ENX), fleroxacin (FLX), pefloxacin (PFX) and pipemidic acid (PPA). The CL conditions of Dy+3-ENX-MnO4−-S2O3−2-HNO3, Dy+3-FLX-MnO4−-S2O3−2-H6P4O13, Dy+3-PFX-MnO4−-S2O3−2-HCl, and Dy+3-PPA-MnO4−-S2O3−2-H2SO4 systems were investigated and optimized. The CL spectra are formed from the narrow characteristic emission of Dy+3 at 482 and 578 nm through the intermolecular energy transfer from the excited SO2* to analyte, followed by intramolecular energy transfer from analyte* to Dy+3. The calibration curves for the four analytes have good linearity. The relative standard deviations (RSDs) are in the range of 1.6–1.9% for 11 determinations of 6.0 × 10−8 g/mL of ENX, FLX, PFX, and PPA. The detection limits (3σ) are in the range of 2.2 × 10−10–6.0 × 10−10 g/mL. The proposed four CL-based methods have high sensitivity, precision and potential capability for the determination of residues of quinolone synthetic antibiotics in foods and biological samples.

Simultaneous voltammetric analysis of tetracycline antibiotics in foods

The new simple, cost effective and well performing differential pulse stripping voltammetry method for the simultaneous determination of antibiotics – oxytetracycline, tetracycline and chlorotetracycline was researched and developed. It depends on the reduction of these compounds at a hanging mercury drop electrode. The samples were extracted from (i) spiked animal feed, and (ii) fresh fish muscle dosed with the drugs. The voltammograms from the drug mixture produced complex, overlapping profiles, and chemometrics methods were applied for calibration modelling. The analytical linear ranges were within 0.02–0.18 μg mL −1 and the corresponding LODs were 3–5 μg L −1 for the three analytes. These values compare well with those from the HPLC and fluorescence methods in the literature. The % relative prediction errors from the verification trials were between 4% and 9% with % Recoveries being 103–107. Also, the % Recoveries of the antibiotics from animal feeds as measured by the new method were comparable with those from the HPLC analysis (85%), i.e. the method is highly competitive, especially as a screening approach.

Erratum to: Antibiotics in food: Legislation and validation of analytical methodologies

The article was published in Vol. 395, issue 4, on pages 877–891. Recently, the authors became aware that Regulation (EEC) no. 2377/90 was repealed by Regulation (EC) no. 470/2009. The correct version of the first row of Table 1 on page 879 is given here: Also, in the text on page 880 three corrections need to be made: Line 2: incorrect: 2377/90, correct: 470/2009. Line 23: “or Annex IVof Regulation 2377/90” should be deleted. Line 43: “Regulation 2377/90 and” should be deleted. The correct reference 31 should read: Council of the European Union (2009) Off J Eur Union L 152:11 Table 1 Main legislative documents of the EU (in chronological order)

Antibiotics in food and environmental samples

Antibiotics in food and environmental samples

Antibiotics in food: Legislation and validation of analytical methodologies

In this paper, European food safety legislation is presented, and special attention is devoted to monitoring residues of veterinary drugs in foodstuffs of animal origin. After a short review of the state of the art of analytical methodology for antibiotic residue analysis, the paper focuses on validation of analytical methods, with Decision 657/2002/EC as reference document. Finally, the main issues of the quality control of the analytical data, i.e. analysis of reference materials and participation in proficiency tests, are briefly addressed.

Applications of capillary electrophoresis to the determination of antibiotics in food and environmental samples

In this paper we review applications of capillary electrophoresis (CE) to the determination of antibiotic residues in food derived from animals and in environmental samples. Although many CE methods have been used to determine antibiotics in the pharmaceutical field (drug quality control or therapeutic monitoring in biological samples), food and environmental applications have been increasing in recent years. Due to the maximum residue limits established by the EU, in Directive 2377/90/EEC, for foodstuffs of animal origin and considering the low levels that can be found in environmental or waste waters or soils, different strategies to increase sensitivity have been developed, including off-line preconcentration, on-line stacking modes to use higher sample volumes, or in-line solid-phase extraction. Also, several detection techniques, such as fluorescence, laser-induced fluorescence, electrochemical detection, or mass spectrometry have been used; the last of these also enables unequivocal identification of the residues, required by Commission Decision 2002/657/EC. All these aspects will be discussed in this paper, in relation to the main groups of antibiotics used in veterinary and human medicine, for which applications in food and environmental samples have been developed by using CE as an efficient alternative to liquid chromatography.

Monoclonal antibody production and immunochemical detection of polyether antibiotics

Polyether antibiotics such as monensin and salinomycin have been primarily used as coccidiostat and growth promoter. Since residues of these antibiotic in food may pose a health risk for sensitive individuals, their use should be carefully monitored. An immunochemical method was developed for the determination of polyether antibiotic using monoclonal antibody (Mab) produced by immunized mice. Conjugates of monensin, salinomycin and laidlomycin were prepared with bovine serum albumin (BSA), keyhole limpet haemocyanine (KLH) and ovalbumin (OVA) by mixed anhydride method and then used as immunogene to produce Mab. Eight hybridoma cell lines were isolated that produced Mabs that competed with polyether antibiotic-protein conjugates in BALB/c-SP2/0 fusion system. Two hybridoma with higher sensitivity, designated as 4G11F and 1C8F1F, were cultured for mass production and then purified from ascites fluid. Antibiotic-protein conjugates were quantitavely analyzed by using the purified Mabs through a competitive enzyme-linked immunosorbent assay (ELISA).

Variations in the microbiology of peritonsillar abscess

Microbiologic studies are routinely performed in the assessment of peritonsillar abscess (PTA). Though the bacterial growth rates of PTA are expected to be uniform due to high accessibility and reasonable sterility, they demonstrate a vast range of results, which is partially explained by the differing culturing methods and incubation times. Our aim was to retrospectively examine the changing features identified in the occurrence of PTA bacterial growth rates over a period of seven years. A retrospective study was undertaken on all cases of PTA admitted from January 1996 to December 2002. Details regarding sex, age and country of birth were obtained. Population data and the maximum residue level (MRL) of antibiotics in food were collected. Bacteriologic studies were analysed for gram stain, aerobic and anaerobic culture results, and also antibiotic sensitivities, if obtained. Four hundred and fifty-seven consecutive hospitalisations due to PTA were identified; 281 patients who had 310 hospitalisations with known results of the microbiologic studies were included. The most common pathogens were Streptococcus pyogenes and Prevotella. A statistically significant escalation was seen in the anaerobic growth rate from 6.8% of cases in 1996 to 37% in 1999. A similar change, though not significant, was noticed with the polymicrobial growth rate. None of the parameters investigated showed any statistically significant influence on this tendency. These results may clarify the immense range of bacterial study results reported, suggest a change in the biologic behaviour of the studied bacteria and direct further research.

Development of a Monoclonal Antibody-Based Broad-Specificity ELISA for Fluoroquinolone Antibiotics in Foods and Molecular Modeling Studies of Cross-Reactive Compounds

A competitive indirect enzyme-linked immunosorbent assay (ciELISA) using monoclonal antibodies (Mabs) having broad specificity for fluoroquinolone (FQ) antibiotics is described. Four FQs, ciprofloxacin (CIP), enrofloxacin (ENR), norfloxacin (NOR), and ofloxacin (OFL), were conjugated to bovine serum albumin for immunogens and to ovalbumin for coating antigens. A Mab C4A9H1 raised against the CIP hapten exhibited high cross-reactivity (35-100%) with 12 of 14 FQs and detected these FQs in a ciELISA below their maximum residue levels (MRLs) with good sensitivity at 50% binding inhibition (IC50). The quantitative structure-activity relationship (QSAR) between Mab C4A9H1 and various FQs by comparative molecular field analysis (CoMFA) showed a high predictive ability with a cross-validation q2 value of 0.866. Using a simple purification process and the broad-specificity ciELISA adapted for analysis of FQs in chicken muscle, chicken liver, honey, shrimp, and whole egg samples demonstrated recoveries of 60-93% for CIP, ENR, NOR, OFL, flumequine, and danofloxacin.

Detection of antibiotics in food: Extraction of fluoroquinolones by DNA

The ability of DNA to extract fluoroquinolones from model solutions and real probes of food was demonstrated and investigated quantitatively. The interaction between fluoroquinolones and different types of DNA was studied by equilibrium dialysis. The first application of this direct approach allowed us to determine binding constants and binding stoichiometries in different conditions. The binding of enrofloxacin to heat-denatured DNA (d-DNA) from herring sperm is pH- and magnesium-dependent; the highest fraction of bound drugs was found at pH 7 and a magnesium concentration of 0.5-1 mM. Results for three types of DNA: d-DNA, double-stranded DNA and single-stranded DNA were compared. The unwound DNA showed almost doubled binding constants and stoichiometries, thus indicating preferable interaction of enrofloxacin with single-strand regions of DNA. The binding of other fluoroquinolones (lomefloxacin, ciprofloxacin, norfloxacin, danofloxacin and sarafloxacin) with d-DNA is very similar to that of enrofloxacin: the binding constants are in the range from 0.94x10(5) to 2.40x10(5) M-1, and the stoichiometries range from 4.1 to 6.9 fluoroquinolone molecules per 100 DNA bases. The binding properties were quantitatively the same for extraction of fluoroquinolones from model aqueous solutions and from liquid food (milk). The results indicate the efficiency of DNA for selective extraction of fluoroquinolones from real samples for further analysis. This selective binding also allows us to consider DNA as a natural receptor for development of analytical techniques for fluoroquinolones.

Determination of Aminoglycosides and Quinolones in Food Using Tandem Mass Spectrometry: A Review

The widespread use of antibiotics in dairy cattle management may result in the presence of antibiotic residues in food. While rapid screening tests are commonly used to detect the presence of antibiotics in food, more accurate chromatographic-mass spectrometric methods combined with tandem mass spectrometry (MS/MS) are required to determine the identity and quantity of the antibiotic present. These methods (HPLC/MS/MS) may have the greatest potential for accomplishing direct multi-residue identifications in complex biological matrices, such as food. This study reviews recent applications of tandem mass spectrometry in the determination of antibiotic residues, such as aminoglycosides and quinolones in food.