Inactivation of penicillin G in milk using hydrogen peroxide.

Milk antibiotic residues have been a public concern in recent years. The Grade A Pasteurized Milk Ordinance mandates that raw Grade A milk will test negative for beta-lactam antibiotic residues before processing. The purpose of this research was to investigate the ability of various levels of peroxide and heat to inactivate penicillin G in raw milk. Whole milk spiked to a mean of 436 +/- 15.1 (standard error of the mean) ppb of potassium penicillin G was treated with hydrogen peroxide at levels of 0.0, 0.09, 0.17, and 0.34%. Samples at each peroxide level (n = 6 per treatment) were treated as follows: 1) incubated at 54.4 degrees C for 3 h, 2) pasteurized at 62.8 degrees C for 30 min, 3) incubated and pasteurized as in treatments 1 and 2, or 4) received no further treatment. A beta-lactam competitive microbial receptor assay was used for quantification of penicillin G. Concentrations of penicillin in selected samples were determined by HPLC for a comparison of test methods. Treatments were evaluated relative to their ability to reduce milk penicillin G levels to below the safe level of 5 ppb. The 0.09% hydrogen peroxide level was ineffective for all treatments. Hydrogen peroxide at 0.17% lowered the mean penicillin G (+/- SEM) from 436 +/- 15.1 to 6 +/- 1.49 ppb using the incubated and pasteurized heat treatment. The 0.34% concentration of hydrogen peroxide was the most effective, inactivating penicillin G to a level well below the safe level of 5 ppb with the pasteurized heat treatment, with or without incubation.

Potential for milk containing penicillin G or amoxicillin to cause residues in calves.

The potential for antibiotic residues in calves from consuming milk containing penicillin G or amoxicillin was investigated. Six calves were fed milk replacer, 6% body weight twice daily, containing 0.293, 2.92, or 5.85 microg of penicillin/ml (ppm) G or 0.25, 1.0, or 2.0 microg of amoxicillin/ml for three consecutive feedings. Urine and blood samples were collected after each feeding. Serum and urine samples were tested with a microbial receptor assay and a microbial growth inhibition assay to indicate potential drug residues. Penicillin G and amoxicillin were detected in the serum and urine of several calves 3 h after drinking spiked milk replacer. Possible violative drug residues in the calves were detected by the microbial growth inhibition assay up to 15 h after drinking spiked milk replacer. Penicillin G, but not amoxicillin, could be detected in urine 24 h after the final feeding of spiked milk replacer. Subsequently, six calves were fed milk replacer containing 11.7 microg of penicillin G/ml (ppm) twice daily, 6% body weight per feeding. Calves were slaughtered 3 h after the final feeding. Mean (+/-SD) concentrations of penicillin G measured by high-pressure liquid chromatography in liver, kidney, muscle, and serum were 0.409 (+/-0.167) microg/g, 0.031 (+/-0.012) microg/g 0.008 (+/-0.002) microg/g, and 0.013 (+/-0.006) mg/ml, respectively. This study indicates that calves fed milk with amoxicillin or penicillin G could possibly have violative residues if slaughtered within 24 h after feeding. Violative drug residues in liver tissue were found in calves slaughtered 3 h after consuming milk replacer containing 11.7 microg of penicillin G/ml (ppm).

Conversion of cephapirin to deacetylcephapirin in milk and tissues of treated animals.

Cephapirin is one of six beta-lactam antibiotics approved for use in the treatment of food-producing animals in the United States. When used for treatment of mastitis by intramammary infusion, it is partially converted to a microbiologically active metabolite identified as deacetylcephapirin (DACEP). The degradation was followed in four cows with naturally acquired mastitis which were treated with cephapirin. DACEP persisted longer than the parent compound in the milk. When a calf was treated with cephapirin by intramuscular injection, the compound was almost completely converted to DACEP in tissues. The deacetyl form must be considered in the determination of residues in treated animals.

Detection of milk antibiotic residues by use of screening tests and liquid chromatography after intramammary administration of amoxicillin or penicillin G in cows with clinical mastitis.

OBJECTIVE: To compare results of 6 commercially available milk antimicrobial screening tests with results of liquid chromatography (LC) when testing milk samples from individual cows treated for mild clinical mastitis by intramammary (IMM) infusion with amoxicillin or penicillin G. ANIMALS: 6 cows with noninduced clinical mastitis: 3 treated by IMM infusion with amoxicillin and 3 treated by IMM infusion with penicillin G. PROCEDURE: Composite milk samples were collected before, during, and after treatment. Samples were assayed by use of the screening tests and their results and those of LC were compared. The LC results were assumed to represent the true result. RESULTS: Results of screening tests compared well with results of LC, with agreement of 94%. Positive screening test results for samples containing drug values below the established tolerance or safe level, as evaluated by LC, were obtained from 2 cows in which abnormal milk, as well as marked increases in composite milk somatic cell count, were observed. With the exception of 1 test in 1 cow, all screening tests had negative results at the end of the labeled milk-withholding time. CONCLUSIONS AND CLINICAL IMPLICATIONS: On the basis of results of the limited sample reported, the screening tests appeared to provide good agreement overall, compared with LC results, when testing milk of individual cows treated by IMM infusion with amoxicillin or penicillin G. Positive screening test results for milk samples containing amoxicillin or penicillin G at values below the established tolerance or safe level, as evaluated by LC, may occasionally be obtained.

Heat resistance of Alicyclobacillus acidoterrestris spores as affected by various pH values and organic acids.

Alicyclobacillus acidoterrestris, a thermoacidophilic sporeformer, has caused spoilage of fruit juices which had been treated with thermal processes intended to commercially sterilize the juice. The objective of this research was to document the effect of pH, acid, and temperature on the heat resistance of spores of three fruit-juice isolates of A. acidoterrestris. The thermal resistance of spores of A. acidoterrestris strains VF, WAC, and IP were studied in a model fruit-juice system composed of 12% glucose and 30 mM of either citric, malic, or tartaric acid, adjusted to selected pH values ranging from 2.8 to 4.0. Decimal reduction times (D values) and inactivation rates were determined. Spores of strains VF and WAC were similarly resistant to heat under acidic conditions, while strain IP spores were less resistant. In the range of pH 2.8 to 4.0, a statistically effect of hydrogen ion concentration on heat resistance was observed at lower temperatures, but not at the higher temperatures, but not at the higher temperatures. For examples, at 91 degrees C and pH 3.1 and 3.7, D values were 31.3 and 54.3 min, respectively, while at 97 degrees C D values at pH 3.1 and 3.7 were 7.9 and 8.8 min, respectively. The type of acid did not significantly affect the heat resistance. The zd values ranged from 5.9 to 10 degrees C, depending on the acid, pH, and the strain. The models generated from this research can be used to determine adequate thermal processes, accounting for the acid type, pH, and temperature, to destroy A. acidoterrestris spores in beverages, since this organism is able to survive the typical hot-fill and hold process (2 min at 88 to 96 degrees C) currently used to process fruit juice.

Procedures used by North Carolina dairies for vitamins A and D fortification of milk.

New research findings have documented fortification errors in fortified milk products all across the US milk industry. Also, the consumption of overfortified bovine milk has led the public to question whether vitamin fortification is safe. Therefore, North Carolina dairies were surveyed to determine vitamin fortification procedures used and to determine differences among these procedures. Of the parameters surveyed, the general conditions under which vitamin preparations were stored, the method used to add vitamin preparations to milk, and the point during processing at which vitamin preparations were added to milk were different among dairies. Forty-six percent of the dairies stored vitamin preparations under refrigerated conditions, and 54% stored vitamin preparations at ambient temperatures. The addition of vitamin preparations to bovine milk was accomplished by metered injection at 64% of the dairies and batch addition techniques at 36%. Vitamin preparations were added before fat content standardization and separation by 23% of the dairy processors; 77% added the vitamin preparations after this point. When dairies were asked at what point they added their vitamin preparations to the milk, nine different answers were given. Although other sources of error could also contribute to inconsistent concentrations of vitamin fortification, differences in fortification procedures may have a large impact upon the problem. The diversity of vitamin fortification practices used in North Carolina may be an indication of nationwide trends.

Ampicillin and amoxicillin residue detection in milk, using microbial receptor assay (Charm II) and liquid chromatography methods, after extra-label administration of the drugs to lactating cows.

OBJECTIVE: A microbial receptor assay method (MRAM; Charm II test) for beta-lactam antibiotics and a liquid chromatography (LC) method with a detection limit of 2 to 5 ppb were evaluated for detection of ampicillin or amoxicillin residues in milk samples from individual cows. DESIGN: The MRAM was compared to the LC in 2 respects. Measured concentrations of drugs were compared, as well as the classification of samples relative to the FDA tolerance value of 10 ppb. ANIMALS: A total of 6 clinically normal lactating Holstein cows were used per drug. PROCEDURE: Ampicillin trihydrate or amoxicillin trihydrate was administered at an extra-label dosage of 22 mg/kg of body weight, IM, once to each of 6 cows/drug. Milk samples were collected at milkings prior to and for 156 hours after drug administration. Drug concentrations in milk samples from individual cows were determined by use of the MRAM and LC tests. Additionally, the classification of milk samples relative to the presence or absence of residues above the FDA tolerance value was determined. Pharmacokinetic analysis was performed on derived milk drug concentrations. RESULTS: Concentration of ampicillin in milk samples from all cows was < 10 ppb by the MRAM and LC methods by the fourth milking (48 hours) after treatment with ampicillin. Values were < 10 ppb by both methods for all cows treated with amoxicillin by the sixth milking (72 hours) after treatment. For individual milk samples, significant differences were found between test methods in the proportion of positive (failing) tests; the MRAM had a higher proportion of presumptive positives. CONCLUSIONS: Even at an extra-label dosage of 22 mg/kg, IM, milk residues > 10 ppb (the FDA tolerance value) were not detected beyond the label milk withholding times for ampicillin (48 hours) and amoxicillin (96 hours). When used for testing milk of individual cows by the control point procedure, the MRAM had a tendency to give presumptive positive test results for milk samples containing < 10 ppb ampicillin or amoxicillin as determined by LC.

Comparison of a radioimmunoassay (Charm II) test with high-performance liquid chromatography for detection of oxytetracycline residues in milk samples from lactating cattle.

A radioimmunoassay test for tetracyclines (Charm II) was compared with high-pressure liquid chromatography (HPLC) for detection of oxytetracycline (OTC) residues in milk samples from individual lactating cows. Oxytetracycline was administered by 1 of 3 routes (IV, IM, or intrauterine) to 21 lactating dairy cows. A total of 292 duplicate milk samples were collected from milkings before and through 156 hours after OTC administration. Concentration of OTC in these samples was determined by use of the Charm II test and an HPLC method with a lower limit of quantitation, approximately 2 ng of OTC/ml. Samples were also classified with respect to presence of OTC residues relative to the FDA safe concentration (< or = 30 ng/ml), using the Charm II (by control point determination) and HPLC methods. There was a significant (P < or = 0.05) difference between test methods in classification of milk samples with respect to presence or absence of OTC at the FDA safe concentration. A total of 48 of the 292 test results (16.4%) did not agree. Using the HPLC test results as the standard with which Charm II test results were compared, 47 false presumptive-violative test results and 1 false presumptive-nonviolative Charm II test result (a sample containing 31 ng of OTC/ml, as evaluated by HPLC) were obtained. The samples with false presumptive-violative Charm II results contained < 30 ng of OTC/ml, as evaluated by HPLC. In some respects, the Charm II test performed appropriately as a screening test to detect OTC residues in milk samples from individual cows.(ABSTRACT TRUNCATED AT 250 WORDS)

Potential for oxytetracycline administration by three routes to cause milk residues in lactating cows, as detected by radioimmunoassay (Charm II) and high-performance liquid chromatography test methods.

Milk antimicrobial residues are a serious concern for the dairy industry. Residues of the tetracycline family of antimicrobials have been reported in market milk by investigators, using radioimmunoassay and microbial receptor technology (hereafter referred to as the Charm II test). In response to these reports, an investigation was conducted to determine the potential of 3 extra-label routes of oxytetracycline (OTC) administration to cause milk residues above the Food and Drug Administration safe value of 30 parts per billion (ppb). Lactating Holstein cows were administered OTC once by use of 1 of 3 routes: IV at 16.5 mg/kg of body weight (n = 6); IM at 11 mg/kg (n = 6); and intrauterine (IU) at 2 g in 500 ml of saline solution/cow (n = 6). Duplicate milk samples were collected at the milking prior to drug administration and for the next 13 milkings at 12-hour intervals. Concentrations of OTC in milk samples were analyzed by use of the Charm II tes for tetracyclines (limit of OTC detection, approx 5 ppb) and were compared with concentrations determined by use of a high-performance liquid chromatography (HPLC) method (lower limit of OTC quantitation, approx 2 ppb). The potential for milk OTC residues above the Food and Drug Administration safe value of 30 ppb after treatment was considerably greater for the IV and IM routes, compared with the IU route.(ABSTRACT TRUNCATED AT 250 WORDS)