Comparison of Rapid Tests Used to Detect Antibiotic Residues in Milk1

1990 ◽  
Vol 53 (2) ◽  
pp. 158-164 ◽  
Author(s):  
GARY F. SENYK ◽  
JOSEPH H. DAVIDSON ◽  
JANICE M. BROWN ◽  
ERIC R. HALLSTEAD ◽  
JOHN W. SHERBON
Keyword(s):  
Bacillus Stearothermophilus ◽  
Raw Milk ◽  
Penicillin G ◽  
Beta Lactam ◽  
Rapid Methods ◽  
Milk Samples ◽  
Beta Lactam Antibiotics ◽  
Rapid Tests ◽  
Combined Data ◽  
Disc Assay

Five rapid methods for detection of antibiotics in milk were compared. The Bacillus stearothermophilus var. calidolactis disc assay was also performed on the same samples. The rapid methods were: Angenics Spot Test, Charm II, Delvotest P, Penzyme Farm, and Penzyme Lab III. Ten antibiotics (penicillin G, cephapirin, cloxacillin, ampicillin, streptomycin, chloramphenicol, erythromycin, novobiocin, tetracycline, and gentamicin) were used individually to spike eight raw milk samples at five levels of antibiotic. Antibiotic levels were chosen that would result in zones of <16 mm, 16 mm, and >16 mm on the disc assay. Only the disc assay, Charm II and Delvotest P were compared on non-beta-lactam antibiotics. A small percentage of milks with no antibiotic added tested positive with the Charm II and Penzyme Lab III. On combined data for penicillin G, cephapirin, and cloxacillin, for which all methods were compared, the percent correctly categorized as pass (below actionable) for the <16 mm zone spiked level, reject or caution at the 16 mm zone level, and reject or caution at the >16 mm zone level were: Angenics 79, 83, 100; Charm II 66, 92, 100; Delvotest P 74, 93, 100; Disc Assay 100, 74, 100; Penzyme Farm 93, 61, 92; Penzyme Lab III 81,78, 100 respectively. In most cases, the rapid methods showed greater apparent sensitivity than the disc assay and did not fail to reject milks spiked with antibiotic in excess of the 16 mm zone level.

scholarly journals Validation Study of a Receptor-Based Lateral Flow Assay for Detection of Beta-Lactam Antibiotics in Milk

2009 ◽  
Vol 92 (3) ◽  
pp. 959-974 ◽  
Author(s):  
Mohamed Abouzied ◽  
Michael Sarzynski ◽  
Aaron Walsh ◽  
Heather Wood ◽  
Mark Mozola
Keyword(s):  
Bovine Milk ◽  
Operating Parameter ◽  
Raw Milk ◽  
Cross Reactivity ◽  
Penicillin G ◽  
Beta Lactam ◽  
Milk Samples ◽  
Beta Lactam Antibiotics ◽  
Cell Counts ◽  
Lactam Antibiotic

Abstract Avalidation study designed tomeet the requirements of the AOAC Research Institute and the U.S. Food and Drug Administration (FDA), Center for Veterinary Medicine, was conducted for a receptor-based, immunochromatographic method (BetaStar US) for detection of beta-lactam antibiotic residues in raw, commingled bovine milk. The assay was found to detect amoxicillin, ampicillin, cephapirin, cloxacillin, and penicillin G at levels below the FDA tolerance/safe levels but above the maximum sensitivity thresholds established by the National Conference on Interstate Milk Shipments. Results of the Part I (internal) and Part II (independent laboratory) dose-response studies using spiked samples were in very close agreement for all five drugs tested, with differences between the Part I and Part II 90/95 sensitivity values ranging from 0 to 1 ppb. The test was able to detect all five drugs at the approximate 90/95 sensitivity levels when present as incurred residues in milk collected from cows that had been treated with the specific drug. Asixth drug, ceftiofur, was found to be undetectable at levels of 500 ppb (as total ceftiofur metabolites from incurred residues in milk samples). The selectivity of the assay was 100, because no false-positive results were obtained in tests of >1000 control milk samples. The assay was found to be applicable to the testing of frozen raw milk samples. Results of ruggedness experiments established the operating parameter tolerances for the BetaStar US assay. Results of cross-reactivity testing established that the assay detects certain other beta-lactam drugs (dicloxacillin and ticarcillin), but it does not cross-react with any of 30 drugs belonging to other classes. Abnormally high bacterial or somatic cell counts in rawmilk produced no interference with the ability of the test to detect beta-lactams at tolerance/safe levels.

Bacillus stearothermophilus Disc Assay for Detection of Inhibitors in Milk: Collaborative Study

1982 ◽  
Vol 65 (5) ◽  
pp. 1208-1214 ◽  
Author(s):  
James W Messer ◽  
James E Leslie ◽  
Gary A Houghtby ◽  
James T Peeler ◽  
Jerald E Barnett ◽  
...  
Keyword(s):  
Bacillus Stearothermophilus ◽  
Collaborative Study ◽  
Raw Milk ◽  
Penicillin G ◽  
Confirmatory Test ◽  
Beta Lactam ◽  
Aoac Method ◽  
Positive Results ◽  
Paper Disc ◽  
Disc Assay

Abstract A 2-part (A and B) collaborative study was conducted on a Bacillus stearothermophilus paper disc (12.7 mm) method to detect residual inhibitors in milk. The 18 participating collaborators assayed raw milk samples spiked with a beta-lactam (penicillin G). Of the 18 collaborators, 14 participated in part A and 16 in part B. Part A demonstrated that either Antibiotic Medium No. 4 or PM Indicator Agar is suitable for use in the assay. The lowest concentration detectable, not significantly different from 100% at the α = 0.05 level, was 0.008 unit/mL with either medium. Part B demonstrated that the sensitivity of the method is equal to that of the current AOAC method (16.131- 16.136). The concentration of beta-lactam detected by 50% of the analysts was 0.003-0.005 unit/mL in this study, compared with 0.005 unit/mL reported in an earlier collaborative study on the current AOAC method. No false positive results were reported in part A or part B. All samples found positive by the confirmatory test in part B were correctly identified as a beta-lactam with commercial Penase discs. The lowest concentration detectable by the method, not significantly different from 100% at the α = 0.05 level, was 0.008 unit/mL. The method was adopted official first action.

Detectability Levels of Four Beta-Lactam Antibiotics in Eight Milk Products Using the AOAC Bacillus stearothermophilus Disc Assay

1986 ◽  
Vol 49 (9) ◽  
pp. 687-690 ◽  
Author(s):  
KATHLEEN T. RAJKOWSKI ◽  
JAMES T. PEELER ◽  
JAMES W. MESSER
Keyword(s):  
Bacillus Stearothermophilus ◽  
Penicillin G ◽  
Beta Lactam ◽  
Milk Products ◽  
Beta Lactam Antibiotics ◽  
Goat’S Milk ◽  
Confirmation Test ◽  
Goat's Milk ◽  
Disc Assay

The 50% dectability level (ED50) of the Bacillus stearothermophilus disc assay in raw, pasteurized whole, protein-fortified lowfat, lowfat, and skim milks, half-and-half, heavy cream and goat's milk was determined for penicillin G, ampicillin, cloxacillin and cephapirin. Results demonstrate a lower level of detectability with PM agar than with A4 agar for ampicillin, cloxacillin and penicillin. Ranges of detection using PM agar at 64°C were 0.0025 to 0.0042 IU/ml (0.0016 to 0.0026 μg/ml) for penicillin G, 0.0021 to 0.0042 μg/ml for ampicillin, 0.0030 to 0.0059 μg/ml for cephapirin and 0.0167 to 0.0334 μg/ml for cloxacillin. Liquid penicillinase is recommended when performing the confirmation test for beta-lactam identification.

Precision Parameters for AOAC Bacillus stearothermophilus Disc Assay Based on FDA Milk Laboratory Quality Assurance 1982–1986 Samples

1989 ◽  
Vol 52 (12) ◽  
pp. 867-870 ◽  
Author(s):  
J. T. PEELER ◽  
J. W. MESSER ◽  
G. A. HOUGHTBY ◽  
J. E. LESLIE ◽  
J. E. BARNETT
Keyword(s):  
Quality Assurance ◽  
Bacillus Stearothermophilus ◽  
Penicillin G ◽  
Test Results ◽  
Inhibitory Substance ◽  
Milk Samples ◽  
Relative Standard ◽  
Standard Deviations ◽  
Estimate Precision ◽  
Disc Assay

Inhibitory substance (antibiotic) test results from State Split Milk Samples were used to estimate precision parameters and to compare antibiotic medium 4 (A4) and PM indicator (PM) agars. Five inhibitory substances (ampicillin, cephapirin, erythromycin, neomycin, and penicillin-G) were tested. Repeatability relative standard deviations (RSDr) ranged from 1.0 to 4.8%, and the reproducibility relative standard deviations (RSDR) ranged from 4.8 to 10.4%. Zone sizes of erythromycin, neomycin, and penicillin-G were significantly larger on PM agar (α = 0.05) than on A4 agar. The reverse was observed for cephapirin. No difference between agars was noted for ampicillin.

Quantitative Estimates of Beta-Lactam Residues in Raw Milk Around a Reference Standard: Collaborative Study

1982 ◽  
Vol 65 (6) ◽  
pp. 1407-1412
Author(s):  
Roy E Ginn ◽  
Ronald A Case ◽  
Vernal S Packard ◽  
Sita R Tatini ◽  
◽  
...  
Keyword(s):  
Bacillus Stearothermophilus ◽  
Collaborative Study ◽  
Raw Milk ◽  
Assay Method ◽  
Zone Size ◽  
Reference Standard ◽  
Beta Lactam ◽  
Quantitative Estimates ◽  
The Difference ◽  
Disc Assay

Abstract A collaborative study was conducted to determine the reliability of a Bacillus stearothermophilus disc assay method for differentiating various concentrations of penicillin in raw milk. Participating laboratories tested 10 different samples (including one negative) in blind duplicate. Triplicate standards were alternated with triplicate unknowns around the periphery of each of 5 different plates. Zone diameters were measured and the difference in zone size of pairs of adjacent standard and unknown samples were analyzed by a paired t-test. Penicillin concentrations 0.003 IU/mL different from the reference concentrations were consistently distinguishable at a 95% confidence level. Such discriminatory power was determined to be possible with as few as 3 plates (9 replicates) per unknown. The method has been adopted official first action.

Quantitative Assay of Beta-Lactam Residues in Raw Milk Using a Disc Assay Method

1982 ◽  
Vol 45 (6) ◽  
pp. 571-573 ◽  
Author(s):  
R. E. GINN ◽  
R. CASE ◽  
V. S. PACKARD ◽  
S. TATINI
Keyword(s):  
Bacillus Stearothermophilus ◽  
Raw Milk ◽  
Technical Skill ◽  
Assay Method ◽  
Reference Level ◽  
Reference Standard ◽  
Beta Lactam ◽  
Quantitative Estimates ◽  
Fixed Reference ◽  
Disc Assay

Numerous methods have been developed to determine presence of antibiotics in raw milk. Until recently, major effort had been placed on qualitative considerations, and primarily for detecting presence of penicillin (beta-lactam) residues. Only one method, the Sarcina lutea Cylinder Plate (CP) procedure, has been modified to provide for quantitative estimates. The CP method is a rather long, tedious test, requiring considerable technical skill. Need for a simpler, faster quantitative method was apparent. This paper describes a method for making quantitative estimates of beta-lactam residues around a fixed reference standard. The method uses Bacillus stearothermophilus in a disc assay test. Quantitative estimates above or below the reference level of antibiotic are computed through a paired-t statistical analysis. The test can be completed within 3 h.

Rapid Screening Assay for Beta-Lactam Antibiotics in Milk: Collaborative Study

1982 ◽  
Vol 65 (5) ◽  
pp. 1186-1192 ◽  
Author(s):  
Stanley E Charm ◽  
Ruey K Chi ◽  
◽  
H Bryant ◽  
M Carson ◽  
...  
Keyword(s):  
Cell Wall ◽  
Control Point ◽  
Collaborative Study ◽  
Rapid Screening ◽  
Penicillin G ◽  
Beta Lactam ◽  
Screening Assay ◽  
Milk Samples ◽  
Beta Lactam Antibiotics ◽  
The Mean

Abstract A 15 min assay for beta-lactam antibiotics has been used by dairies for several years as a screening procedure for testing milk tankers before they unload. The test is based on a competition between 14C-penicillin and beta-lactam antibiotics in the milk samples for sites on a microbial cell wall that specifically binds beta-lactam. In a collaborative study, 11 laboratories correctly distinguished 10 coded zero penicillin G samples and 10 coded 0.01 IU/mL samples. The proposed test is qualitative, positive or negative, and can detect the presence of beta-lactam antibiotics at the 0.01 IU/mL level. The control point for determining positive or negative samples is more than 3 standard deviations from the mean of 0.01 IU/mL. The method has been adopted official first action.

Validation Study of the BetaStar® Plus Lateral Flow Assay for Detection of Beta-Lactam Antibiotics in Milk

2012 ◽  
Vol 95 (4) ◽  
pp. 1211-1221 ◽  
Author(s):  
Mohamed Abouzied ◽  
Dana Driksna ◽  
Coilin Walsh ◽  
Michael Sarzynski ◽  
Aaron Walsh ◽  
...  
Keyword(s):  
Validation Study ◽  
Bovine Milk ◽  
Operating Parameter ◽  
Raw Milk ◽  
Cross Reactivity ◽  
Penicillin G ◽  
Beta Lactam ◽  
Beta Lactam Antibiotics ◽  
Cell Counts ◽  
Lactam Antibiotic

Abstract A validation study designed to meet the requirements of the AOAC Research Institute and the U.S. Food and Drug Administration, Center for Veterinary Medicine (FDA/CVM) was conducted for a receptor and antibody-based, immunochromatographic method (BetaStar® Plus) for detection of beta-lactam antibiotic residues in raw, commingled bovine milk. The assay was found to detect amoxicillin, ampicillin, ceftiofur, cephapirin, cloxacillin, and penicillin G at levels below the FDA tolerance/safe levels, but above the maximum sensitivity thresholds established by the National Conference on Interstate Milk Shipments (NCIMS). Results of the Part I (internal) and Part II (independent laboratory) dose-response studies employing spiked samples were in close agreement. The test was able to detect all six drugs at the approximate 90/95% sensitivity levels when presented as incurred residues in milk collected from cows that had been treated with the specific drug. Selectivity of the assay was 100%, as no false-positive results were obtained in testing of 1031 control milk samples. Results of ruggedness experiments established the operating parameter tolerances for the BetaStar Plus assay. Results of cross-reactivity testing established that the assay detects certain other beta-lactam drugs (dicloxacillin and ticarcillin), but it does not cross-react with any of 30 drugs belonging to other classes. Abnormally high bacterial or somatic cell counts in raw milk produced no interference with the ability of the test to detect beta-lactams at tolerance/safe levels.

scholarly journals Evolution and resistance expression of MRSA. Evaluation of beta-lactam antibiotics against a set of isogenic strains with different types of phenotypic expression.

1995 ◽  
Vol 42 (4) ◽  
pp. 517-524 ◽  
Author(s):  
K Asada ◽  
Y Inaba ◽  
E Tateda-Suzuki ◽  
K Kuwahara-Arai ◽  
T Ito ◽  
...  
Keyword(s):  
Meca Gene ◽  
Phenotypic Expression ◽  
Aminoglycoside Antibiotic ◽  
Penicillin G ◽  
Binding Affinities ◽  
Penicillin Binding Protein ◽  
Beta Lactam ◽  
Beta Lactam Antibiotics ◽  
Mrsa Infection ◽  
Resistance Expression

Methicillin-resistant Staphylococcus aureus (MRSA) has two mechanisms of resistance to beta-lactam antibiotics; one is mediated by mecA gene expression, and the other by penicillinase production. It has been generally accepted in the clinical field that beta-lactam antibiotics are not the drugs of choice for MRSA infection. In this report, however, ampicillin and penicillin G were shown to have relatively good activity against MRSA if combined with a beta-lactamase inhibitor, sulbactam. These beta-lactam antibiotics were found to have relatively high binding affinities to PBP2', the mecA-encoded MRSA-specific penicillin-binding protein. The possible therapeutic application of sulbactam/ampicillin against MRSA infection in combination with arbekacin, an aminoglycoside antibiotic newly developed and introduced into clinical use in Japan, is discussed.

scholarly journals A metallo-beta-lactamase with both beta-lactamase and ribonuclease activity is linked with traduction in giant viruses

2019 ◽  
Author(s):  
Philippe Colson ◽  
Lucile Pinault ◽  
Said Azza ◽  
Nicholas Armstrong ◽  
Eric Chabriere ◽  
...  
Keyword(s):  
Acanthamoeba Castellanii ◽  
Deep Ocean ◽  
Penicillin G ◽  
Beta Lactamase ◽  
Beta Lactam ◽  
Strong Activity ◽  
Beta Lactam Antibiotics ◽  
Double Stranded Dna ◽  
Giant Viruses ◽  
Hydrolysis Of

ABSTRACTEnzymatic proteins with a metallo-beta-lactamase (MBL) fold have been essentially studied in bacteria for their activity on beta-lactam antibiotics. However, the MBL fold is ancient and highly conserved, and these proteins are capable of cleaving a broad range of substrates. It has recently been shown that MBLs are present in a wide array of cellular organisms, including eukaryotes and archaea. We show here that Tupanvirus deep ocean, a giant virus, also encodes a protein with a MBL fold. Phylogeny showed its clustering with transfer ribonucleases (RNases) and the presence of orthologs in other giant viruses, mainly those harboring the largest sets of translation components. In addition, it suggests an ancient origin for these genes and a transfer between giant viruses and Acanthamoeba spp., a host of many giant viruses. Biologically, after its expression in Escherichia coli, the tupanvirus protein was found to hydrolyse nitrocefin, a chromogenic beta-lactam. We also observed an hydrolysis of penicillin G (10 μg/mL) and detected the metabolite of penicillin G hydrolysis, benzylpenilloic acid. This was inhibited by sulbactam, a beta-lactamase inhibitor. In addition, we tested the degradation of single-stranded DNA, double-stranded DNA, and RNAs, and observed a strong activity on RNAs from seven bacteria with G+C varying from 42% to 67%, and from Acanthamoeba castellanii, the tupanvirus host. This was not inhibited by sulbactam or ceftriaxone. RNase activity was estimated to be 0.45±0.15 mU/mg using a fluorescence-based assay. Our results still broaden the range of hosts of MBL fold proteins and demonstrate that such protein can have dual beta-lactamase/nuclease activities. We suggest that they should be annotated according to this finding to avoid further confusion.

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