Detection of Penicillin G and its Benzylpenicilloyl (BPO) - Derivatives in Cow Milk and Serum by means of an ELISA

1985 ◽  
Vol 48 (1) ◽  
pp. 59-62 ◽  
Author(s):  
PETER ROHNER ◽  
MELCHIOR SCHÄLLIBAUM ◽  
JACQUES NICOLET
Keyword(s):  
Plasma Level ◽  
Bacillus Stearothermophilus ◽  
Competitive Elisa ◽  
Cow Milk ◽  
Penicillin G ◽  
Biological Assay ◽  
Antibiotic Residues ◽  
Milk Samples ◽  
Bulk Milk ◽  
Long Time

Pharmacokinetic characteristics of benzylpenicillin and its benzylpenicilloyl (BPO)-derivatives were studied in serum and milk of health cows, using a classical biological assay (Sarcina lutea test) and a competitive ELISA for BPO detection. The plasma level and passage into milk was determined after intramuscular administration of penethamate-hydroiodide and benzylpenicillin-procaine. In serum of cows receiving penethamate hydroiodide, BPO seemed to persist for a rather long time; the reason for this observation was not clarified. The effect of local (intramammary) application of penicillin G was followed with milk from cows having healthy and mastitic quarters. In all cases, it was found that BPO was not excreted any longer than active penicillin G in milk. In a further survey, 1015 bulk milk samples from two large dairy regions were examined with the ELISA and a biological assay using Bacillus stearothermophilus var. calidolactis. None of the samples showed detectable BPO or antibiotic residues. It is concluded that milk containing inactive penicillin derivatives, like BPO, is not an important source to cause allergies.

Comparison of Microbiological Methods for Detecting Penicillin and Penicillin-Like Antibiotic Residues in Milk

1984 ◽  
Vol 47 (8) ◽  
pp. 604-610 ◽  
Author(s):  
J. N. GELETA ◽  
H. H. BRYANT ◽  
J. E. HEAVNER ◽  
J. J. COLAIANNE ◽  
G. G. CARTER ◽  
...  
Keyword(s):  
Bacillus Stearothermophilus ◽  
Post Treatment ◽  
Penicillin G ◽  
Antibiotic Residues ◽  
Withdrawal Time ◽  
Water Dispersible ◽  
Milk Samples ◽  
Lactating Cows ◽  
Microbiological Methods ◽  
Specific Reactions

Five microbiological methods for detecting residues of penicillin and penicillin-like antibiotics were compared, and the relative incidence of residues in whole milk samples was determined. Five groups of 10 lactating cows each and five commercial antibiotic products (penicillin G in oil, penicillin G in a water-dispersible oil base, cloxacillin, cephapirin and ampicillin) used for the treatment of mastitis were studied. Milk samples collected from the cows were tested for antibiotic content by the Sarcina lutea or Bacillus subtilis cylinder plate, International Dairy Federation-Association of Official Analytical Chemists (IDF-AOAC) Bacillus stearothermophilus ATCC 10149 disc plate, Delvotest-P, Antibiotic Residue Branch (ARB) B. stearothermophilus C 953 quantitative disc plate and ARB B. stearothermophilus C 953 quantitative cylinder plate methods. All samples tested by the original S. lutea or B. subtilis cylinder plate methods, with the exception of the penicillin G in oil product (Uni-Biotic), were free of antibiotic residues and showed no inhibitors beyond the labeled milk withdrawal time. The newer, more sensitive official IDF-AOAC method, Delvotest-P and both ARB B. stearothermophilus quantitative disc and cylinder plate methods detected measurable amounts of cephapirin and penicillin G in the post-treatment milk for one to three additional milkings beyond the labeled milk discard time. Cloxacillin, as determined by the ARB B. stearothermophilus cylinder plate method, was generally depleted by 48 h. The excretion rate of ampicillin product coincided with the labeled milk discard time. The level of detection of both disc plate methods and Delvotest-P for cephapirin and ampicillin was comparable. Variations in the concentration in milk and the persistence of two penicillin G products were probably due to differences in the formulation and the dosing regimen. Inhibitory substances and non-specific reactions were observed with the IDF-AOAC disc method, the Delvotest-P and both ARB B. stearothermophilus methods in some control and post-treatment milk samples. These results are consistent with reports of other investigators and suggest that the use of more sensitive procedures for monitoring antibiotic residues in milk may result, for some infusion preparations, in an increased incidence of violative residues of penicillin G and cephapirin.

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.

scholarly journals Assessment of ceftiofur residues in cow milk using commercial screening test kits

2019 ◽  
Vol 6 (1) ◽  
pp. e000329 ◽  
Author(s):  
Luc Durel ◽  
Guglielmo Gallina ◽  
Terence Pellet
Keyword(s):  
Screening Test ◽  
Generation Cephalosporin ◽  
Operating Conditions ◽  
Cow Milk ◽  
Screening Tests ◽  
Rapid Screening ◽  
Beta Lactam ◽  
Withdrawal Time ◽  
Milk Samples ◽  
Bulk Milk

Ceftiofur, a third-generation cephalosporin, is one of the most used antibiotics in dairy industry. Intramuscular injection of 1 mg/kgBW ceftiofur hydrochloride (HCl) generally results in 0 hour withdrawal time for the milk in dairy cows. Nevertheless, farmers and dairy processors occasionally complain about ceftiofur-based products in case of positive result to a commercial rapid screening test for the presence of violative residues of antimicrobials (inhibitors) in the bulk milk tank. Six lactating cows were injected with a 50 mg/ml ceftiofur HCl-based product at the dosage regimen of 1 mg/kg, intramuscularly, once a day, for five consecutive days, as per label. Milk samples were then collected just before the very last injection (T0) and then at 12, 24, 36, 48, 60, 72, 84 and 96 hours after the last injection. Individual milk samples were tested using three commercial screening test kits for inhibitor residues: DelvotestSP NT, SNAP Beta-Lactam ST Plus and ROSA MRL Beta-Lactam Test. Since bulk tank is screened in real operating conditions, samples were also diluted to 1:4, 1:10 and tested again. For the Delvotest SP NT, which lowest detected concentration is close the MRL of the ceftiofur (100 µg/kg), all results were negative. For the ROSA MRL Beta-Lactam Test and the SNAP Beta-Lactam ST Plus, several samples yielded positive and doubtful results at T0 and T12. However, after dilution to 1:10, all results were negative. Consequently, when used as officially instructed, the tested 50 mg/ml ceftiofur HCl-based injectable veterinary products are safe, and milk should be free of violative residues of ceftiofur. With consideration to the low specificity and the low positive predictive value of commercial screening tests, positive reactions of the bulk milk should be interpreted as false positive or another risky usage of β-lactam-based medicines in the farm must be investigated.

A Practical Evaluation of the Delvotest P Multi Plate Test in Screening Raw Milk for Antibiotics

1986 ◽  
Vol 49 (11) ◽  
pp. 868-870 ◽  
Author(s):  
LYSE LAROCQUE ◽  
G. A. NEVILLE
Keyword(s):  
Bacillus Subtilis ◽  
Staphylococcus Epidermidis ◽  
Raw Milk ◽  
The Other ◽  
Penicillin G ◽  
Antibiotic Residues ◽  
Plate Test ◽  
Milk Samples ◽  
Practical Evaluation ◽  
Bacillus Subtilis Atcc

The Delvotest P Multi plate test was evaluated by screening 100 milk samples for total antibiotic residues (penicillin G, streptomycin and neomycin). The samples were taken in conjunction with an antibiotic depletion study in milk derived from six cows treated with a multiple antibiotic, intramammary infusion product. Within the limits of sensitivity of the Delvotest, only penicillin G persisted in milk samples taken beyond 60 h, whereas in some samples, the other antibiotics appeared to be depleted as early as 48 h. More sensitive tests, however, detected neomycin (Staphylococcus epidermidis, ATCC 12228) in 50% of samples taken at 60 h and streptomycin (Bacillus subtilis, ATCC 6633) at 14.5 d after discontinuation of infusion.

scholarly journals βeta-lactam and Tetracycline Antibiotic Residues in Cow Milk in the Constantine region, Algeria

2021 ◽  
Vol 53 (3) ◽  
Author(s):  
Noureddine Zeghilet ◽  
Brahim Bouchoucha ◽  
Omar Bouaziz
Keyword(s):  
Milk Sample ◽  
Dairy Industry ◽  
Raw Milk ◽  
Control Programme ◽  
Cow Milk ◽  
Antibiotic Residues ◽  
Pasteurized Milk ◽  
Tetracycline Antibiotic ◽  
Milk Samples ◽  
Antibiotics Residues

The aim of the present study was to investigate the βeta-lactam and tetracycline antibiotic residues in cow milk samples. A total of 122 samples of cow milk were collected from raw milk collectors (109 samples) and from a reconstituted pasteurized milk sales clerk (13 samples) in the Constantine region, Algeria and examined using the ΒetaStar Combo screening kit (Neogen, USA). Results indicates that 13 samples (10.66%) were positive for antibiotics residues: 12 (9.84%) for βeta-lactams (ten (8.20%) raw and two (1.64%) pasteurized milk samples) and only one (0.82%) for tetracyclines in a raw milk sample. It is evident that the Algerian consumer is not sheltered from the danger of antibiotic residues in milk and these inhibitor residues should constitute a constant concern for the dairy industry in Algeria. A control programme should be established.

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.

Detection of Antibiotic Residues in Consumer Milk Supplies in North America using the Charm Test II Procedure

1988 ◽  
Vol 51 (8) ◽  
pp. 632-633 ◽  
Author(s):  
D. L. COLLINS-THOMPSON ◽  
D. S. WOOD ◽  
I. Q. THOMSON
Keyword(s):  
North America ◽  
Bacillus Stearothermophilus ◽  
Lower Sensitivity ◽  
Antibiotic Residues ◽  
Comparative Tests ◽  
Milk Samples ◽  
Assay Procedure ◽  
Antibiotics Residues ◽  
Disc Assay

Two hundred and fourteen consumer milk samples from across North America were examined for antibiotics residues by means of the Bacillus stearothermophilus disc assay (Charm test) and the competitive isotopic (Charm Test II) procedure. Of the 174 samples taken from 16 states, 150 results were positive for one or more antibiotics. The greatest number of positives were sulphamethazine, 82, and tetracyline, 48. Candian samples, 40, also showed the same problem related to tetracycline, 12, and sulphamethazine residues, 12. The Bacillus stearothermophilus disc assay procedure was unable in most cases to detect these residues possibly due to the lower sensitivity of this test. Further comparative tests between Charm Test II and other methods of similar sensitivity are recommended to confirm these findings.

Overmilking causes deterioration in teat-end condition of dairy cows in late lactation

2013 ◽  
Vol 80 (3) ◽  
pp. 344-348 ◽  
Author(s):  
J Paul Edwards ◽  
Bernadette O'Brien ◽  
Nicolas Lopez-Villalobos ◽  
Jenny G Jago
Keyword(s):  
Milk Production ◽  
Flow Rate ◽  
Cow Milk ◽  
Udder Health ◽  
Milk Samples ◽  
Bulk Milk ◽  
Milk Flow ◽  
Holstein Friesian ◽  
Mixed Age ◽  
Friesian Cows

The objective of the study was to determine the effect of varying degrees of overmilking on teat-end hyperkeratosis, milk production variables and indicators of udder health during late lactation. This was examined by assessing the effect of four end-of-milking criteria on 181 spring-calving, mixed-age Holstein-Friesian cows, at an average 217±24 d in milk, over a six-week period. The four treatments were: remove cluster once milk flow rate fell to 0·2 kg/min plus 5 s (Ovr0), plus 120 s (Ovr2), plus 300 s (Ovr5), and plus 540 s (Ovr9). Daily measurements included individual cow milk yield, milking duration, overmilking duration, maximum milk flow rate, milk flow rate at cluster removal and the number of cluster re-attachments. Individual cow bulk milk samples were collected weekly at AM and PM milkings to determine composition (fat, protein and lactose) and somatic cell count (SCC; AM only). Teat-end hyperkeratosis score was assessed at weeks 0, 3, 5 and 6. At week 6 mean teat-end hyperkeratosis score of the Ovr2 treatment was not greater than Ovr0, whilst Ovr5 was greater than Ovr2 and Ovr9 was greater than Ovr5 and Ovr2. Milk production, milking characteristics and SCC were not different between treatments, except milking duration and milk flow rate at cluster removal. However, higher teat-end hyperkeratosis scores may have a longer-term impact on indicators of udder health if teat-end condition reaches severe levels. Results indicate that to minimise changes in teat-end condition overmilking should be limited to 2 min, which has implications for milking management in large parlours not fitted with automatic cluster removers.

scholarly journals Determination and Human Health Risk Assessment of Antibiotic Residue in Cow Milk in Bangladesh

2020 ◽  
Author(s):  
Md. Rahman ◽  
Mohammad Hassan ◽  
Sharmin Chowdhury
Keyword(s):  
Risk Analysis ◽  
Health Risk ◽  
Human Life ◽  
Hazard Quotient ◽  
Human Health Risk ◽  
Raw Milk ◽  
Cow Milk ◽  
Antibiotic Residues ◽  
Veterinary Antibiotics ◽  
Milk Samples

Consumption of milk contaminated with antibiotic residues above the maximum residue limit (MRL) causes direct toxicity to humans and the development of superbugs that leads to the failure of antibiotic therapy and threatens human life. Besides, long-duration exposure might alter the nature of gut microflora results in the enhancement of many diseases. Therefore, we examined 300 raw and processed packet milk samples using thin layer chromatography (TLC) and ultra high performance liquid chromatography (UHPLC) method against five veterinary antibiotics and assessed the risk for consumers in Chattogram, Bangladesh. Risk analysis was calculated by using hazard quotient on the basis of 165 ml per capita milk consumption. We found a total of 7% prevalence of antibiotic residues in raw milk samples which were higher (8%) in individual samples than the pooled samples (4%). However, we did not find any processed packet milk samples as positive. The mean concentration of oxytetracycline residue was detected 61.29 µg/l and amoxicillin was 124 µg/l in individual milk samples. Risk analysis showed the hazard quotient values of 0.0056 for oxytetracycline and 0.0017 for amoxicillin residue which was confirmed that, no significant health risk associated with the consumption of milk produced and marketed in the study area. Our study might fill the gap of knowledge to measure the safety status of milk regarding public health issues.

scholarly journals HPLC-DAD multi-residue method for determination of florfenicol, penicillin and tetracycline residues in raw cow milk

2021 ◽  
Vol 2 (3) ◽  
pp. 01-08
Author(s):  
Ali Jaber ◽  
Zeinab Zahreddine ◽  
Simon Abou Haidar ◽  
Chadi Hosri ◽  
Ghassan Ibrahim ◽  
...  
Keyword(s):  
High Performance ◽  
Solid Phase ◽  
Raw Milk ◽  
Reversed Phase ◽  
Cow Milk ◽  
Antibiotic Residues ◽  
Milk Samples ◽  
Antibiotics Residues ◽  
Tetracycline Residues

The existence of antibiotic residues in edible products constitutes a health problem to the consumers. Reversed-phase high-performance liquid chromatography with diode array detection (HPLC–DAD) was optimized and validated for the simultaneous determination of florfenicol (FF), penicillin (PE), and tetracycline (TC) residues in dairy raw milk samples. The determination of these antibiotics was carried out on HP-ODS Hypersil C18 (5μm, 125*4 mm) column at a flow rate (1mL/min) and temperature (35 ⁰C). The extraction method includes deproteinization of the milk sample followed by a solid-phase extraction (SPE) clean-up. The method was validated according to the European Commission Decision 2002/657/EC and the International Conference of Harmonization Guidelines. The recoveries for the studied antibiotics ranged from 82–111.54 % making the method suitable for performing routine analysis. The proposed method was applied for the analysis of antibiotic residues in 50 dairy raw milk samples collected from many regions in Lebanon. The results showed the occurrence of these antibiotics residues in milk collected from different Lebanese regions. The numbers indicate that 22 % of milk samples were found to be positive for FF, 42 % for PE, and 28 % for TC residues.

Sign in / Sign up

Export Citation Format

Share Document