Strategies for the screening of antibiotic residues in eggs: comparison of the validation of the classical microbiological method with an immunobiosensor method
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Efficient screening methods are needed to control antibiotic residues in eggs. A microbiological kit (Explorer® 2.0 test (Zeu Inmunotech, Spain)) and an immunobiosensor kit (Microarray II (AM® II) on Evidence Investigator™ system (Randox, UK)) have been evaluated and validated for screening of antibiotic residues in eggs, according to the European decision EC/2002/657 and to the European guideline for the validation of screening methods. The e-reader™ system, a new automatic incubator/reading system, was coupled to the Explorer 2.0 test. The AM II kit can detect residues of six different families of antibiotics in different matrices including eggs. For both tests, a different liquid/liquid extraction of eggs had to be developed. Specificities of the Explorer 2.0 and AM II kit were equal to 8% and 0% respectively. The detection capabilities were determined for 19 antibiotics, with representatives from different families, for Explorer 2.0 and 12 antibiotics for the AM II kit. For the nine antibiotics having a maximum residue limit (MRL) in eggs, the detection capabilities CCβ of Explorer 2.0 were below the MRL for four antibiotics, equal to the MRL for two antibiotics and between 1 and 1.5 MRLs for the three remaining antibiotics (tetracyclines). For the antibiotics from other families, the detection capabilities were low for beta-lactams and sulfonamides and satisfactory for dihydrostreptomycin (DHS) and fluoroquinolones, which are usually difficult to detect with microbiological tests. The CCβ values of the AM II kit were much lower than the respective MRLs for three detected antibiotics (tetracycline, oxytetracycline, tylosin). Concerning the nine other antibiotics, the detection capabilities determined were low. The highest CCβ was obtained for streptomycin (100 µg kg-1).Keywords:
Maximum Residue Limit
Guideline
Dihydrostreptomycin
Efficient screening methods are needed to control antibiotic residues in eggs. A microbiological kit (Explorer® 2.0 test (Zeu Inmunotech, Spain)) and an immunobiosensor kit (Microarray II (AM® II) on Evidence Investigator™ system (Randox, UK)) have been evaluated and validated for screening of antibiotic residues in eggs, according to the European decision EC/2002/657 and to the European guideline for the validation of screening methods. The e-reader™ system, a new automatic incubator/reading system, was coupled to the Explorer 2.0 test. The AM II kit can detect residues of six different families of antibiotics in different matrices including eggs. For both tests, a different liquid/liquid extraction of eggs had to be developed. Specificities of the Explorer 2.0 and AM II kit were equal to 8% and 0% respectively. The detection capabilities were determined for 19 antibiotics, with representatives from different families, for Explorer 2.0 and 12 antibiotics for the AM II kit. For the nine antibiotics having a maximum residue limit (MRL) in eggs, the detection capabilities CCβ of Explorer 2.0 were below the MRL for four antibiotics, equal to the MRL for two antibiotics and between 1 and 1.5 MRLs for the three remaining antibiotics (tetracyclines). For the antibiotics from other families, the detection capabilities were low for beta-lactams and sulfonamides and satisfactory for dihydrostreptomycin (DHS) and fluoroquinolones, which are usually difficult to detect with microbiological tests. The CCβ values of the AM II kit were much lower than the respective MRLs for three detected antibiotics (tetracycline, oxytetracycline, tylosin). Concerning the nine other antibiotics, the detection capabilities determined were low. The highest CCβ was obtained for streptomycin (100 µg kg-1).
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Guideline
Dihydrostreptomycin
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Virulence factor
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Acute gastroenteritis is common infectious disease in community in adults.This work represents an update of 'Clinical guideline for the diagnosis and treatment of gastrointestinal infections' that was developed domestically in 2010.The recommendation of this guideline was developed regarding the following; epidemiological factors, test for diagnosis, the indications of empirical antibiotics, and modification of antibiotics after confirming pathogen.Ultimately, it is expected to decrease antibiotic misuse and prevent antibiotic resistance.
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The effect of the antibiotic dihydrostreptomycin on the phagocytic and bactericidal ability of peritoneal macrophages obtained from mice has been investigated. In subliminal concentrations which did not influence the bacterial growth, the drug caused macrophages to ingest and kill bacteria ( Escherichia coli ) at a higher rate than did macrophages without antibiotic. The differences for phagocytosis and intracellular killing of E. coli with and without a subliminal amount of dihydrostreptomycin were statistically significant. Macrophages pretreated with the antibiotic did not demonstrate any enhancement of phagocytosis.
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An analytical method was developed for the determination of streptomycin and dihydrostreptomycin in honey using hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS). The streptomycin and dihydrostreptomycin residues in the test samples were extracted with 20 g/L trichloroacetic acid aqueous solution (including 50 mmol/L phosphate, pH 6.8) and cleaned on an Oasis HLB solid phase extraction column. The products were separated on a SIELC Obelisc R column with gradient elution using 0.5% (v/v) formic acid aqueous solution and acetonitrile as mobile phases. Streptomycin and dihydrostreptomycin were detected by liquid chromatography-tandem mass spectrometry in the positive ion mode using the external standard method. Under the optimal conditions, streptomycin and dihydrostreptomycin showed good linearity (r>0.99) in the range of 2.5-100 μg/L. The LOD and LOQ of the method was 2.0 μg/kg and 5.0 μg/kg, respectively. The spiked recoveries of the analytes from blank honey samples at the three levels of 5.0, 20.0 and 100.0 μg/kg were in the range of 86.9%-113.2% with the relative standard deviations less than 10%. With the advantages of convenience, rapidity, sensitivity and good repeatability, the method is suitable for the detection of streptomycin and dihydrostreptomycin in honey.
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OBJECTIVE:To strengthen supervision on the use of antibiotics and to promote rational drug use.METHODS:By analyzing the common pathogenic bacteria in operative site injections,a corresponding guideline for the preventive use of antibiotics was established.RESULTS CONCLUSION:The guideline for prophylactic use of antibiotics established according to our own condition serves as basis for the rational use of antibiotics before operation.
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Competitive exclusion (CE) cultures may offer alternatives to antimicrobial agents for disease prophylaxis in poultry. To avoid potential transfer of antibiotic resistance, safe and effective CE cultures must, by necessity, be highly sensitive to antimicrobial residues. The following studies evaluated the effect of maternal administration of selected antibiotics on the establishment of a licensed CE culture, PREEMPT. Selected antibiotics were administered to actively laying hens for a period of 7 days (experiment 1) or 9 days (experiment 2) in drinking water [sulfadimethoxine (0.05%), enrofloxacin (0.005%), and tylosin tartrate (0.05%)] or feed (sulfadimethoxine with ormetoprim, 250 ppm). In experiment 1, fertile eggs were collected daily and subjected to bioassay for detectable antimicrobial residues in yolk. Antimicrobial residues were not detected during the 7 days of treatment or the subsequent 3 days following cessation of treatment in the control, sulfadimethoxine, sulfadimethoxine with ormetoprim, or tylosin treatment groups. However, detectable residues were observed in eggs derived from enrofloxacin-treated hens on days 6 and 7 during antibiotic administration and also on days 2 and 3 post-antibiotic administration. In experiment 2, antimicrobial residues were also only detected in yolks from hens treated with enrofloxacin. Residue detection occurred on days 2-6 of antibiotic administration, on day 9 of antibiotic administration, on days 1-3 post-antibiotic administration, and also on day 7 post-antibiotic administration. A subset of eggs from each experimental group, corresponding to days 2-6 of antibiotic administration, days 4-6 post-antibiotic administration, and days 14-16 post-antibiotic administration, were pooled for incubation, and chicks hatched from these pools of fertile eggs were treated with PREEMPT at hatch. When 48-h cecal propionate concentrations were used as an index of culture establishment, reduced (P < 0.05) efficacy was observed only in chicks derived from enrofloxacin-treated hens at either collection period. Although several antibiotics do not appear to produce detectable egg residues or interfere with CE culture establishment, these data suggest that chicks derived from enrofloxacin-treated hens may not be candidates for safe and effective CE culture treatment.
Tylosin
Sulfadimethoxine
Dihydrostreptomycin
Tilmicosin
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Antibiotics used in food producing-animals may cause residual problems in food in terms of public health. This situation can lead to serious problems in terms of human health. Raw milk is one of the foods that are likely to contain antibiotics, even in trace amounts. This study aimed to determine the residue levels of commonly used antibiotics of raw goat's milk samples offered for sale in Ankara, Çankırı and Kırıkkale. One active ingredient was selected from the five most commonly used antibiotic groups in animals and it determined the residue levels of these substances in milk. For this purpose, within one year and in two different periods, 150 raw goat milks analyzed in terms of antibiotics using HPLC method. The values for enrofloxacin, sulfamethoxazole, tylosin, penicillin G and oxytetracycline were 7.9, 9.7, 11.5, 5.4 and 7.3 minutes for retention times, 1.47, 0.8; 7.51; 2.69 and 8.89 µg/L for limit of detection (LOD) and 4.47, 2.44, 22.78, 8.16 and 26.96 µg/L for limit of quantification (LOQ), respectively. No antibiotic residues were detected in the goat milk samples. It is predicted that the higher resistance of goats to diseases compared to other milk-producing animals, and therefore the lower use of antibiotics in these animals, leads to this result. The findings obtained as a result of this study are valuable in terms of public health. It is important that no antibiotic residues are found in the analyzes.
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Residue (chemistry)
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The effect of the parenteral administration of aminoglycoside antibiotics on the blood calcium concentration in dairy cows was investigated. Gentamicin was tested in vitro in blood drawn from cows, dihydrostreptomycin was tested in nonlactating cows and neomycin was tested in postpartum cows. The total and bound calcium fractions were significantly reduced by all three antibiotics. No change occurred in the unbound calcium fractions. Caution is advised in the use of these drugs in postpartum cows, especially those with a history of milk fever.
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Neomycin
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Neisseria gonorrhoeae
Neisseria
Pathogenic bacteria
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