The efficacy of flunixin alone and together with enrofloxacin in treatment of experimental Escherichia coli mastitis was compared using six cows. The cross-over study design was used. Pharmacokinetics of flunixin and enrofloxacin were also studied in these diseased cows. The response of each cow was similar after the first and second challenge and the individual reaction seemed to explain the severity of clinical signs. The most important predictive factor for outcome of E. coli mastitis was a heavy drop in milk yield. Treatment with enrofloxacin and flunixin enhanced elimination of bacteria, but the difference from those receiving flunixin alone was not significant. Two cows, which had received no antimicrobial treatment (Group 1), were killed on day 4 postchallenge. One cow was killed after the first and the other after the second challenge. Cows receiving combination therapy produced 0.9 L more milk per day during the study period than cows which had only received flunixin (P < 0.05). Based on our findings, antimicrobial treatment might be beneficial in the treatment of high-yielding cows in early lactation. The absorption of enrofloxacin was delayed after subcutaneous administration, the mean apparent elimination half-life being about 23 h, whereas after i.v. administration elimination t(1/2) was only 1.5 h. The majority of the antimicrobial activity in milk originated from the active metabolite, ciprofloxacin, which could be measured throughout the 120-h follow-up period after the last subcutaneous administration. No differences were present in the pharmacokinetic parameters of flunixin between treatment groups: mean elimination half-life was 5.7-6.2 h, volume of distribution 0.43-0.49 L/kg and clearance 0.13-0.14 L h/kg. No flunixin or merely traces were detected in milk: one of the three cows had a concentration of 0.019 mg/L 8 h after administration.

A microbiological method was developed for group level identification of antibiotics in incurred kidney and muscle samples from cattle and pigs. The method was composed of six test bacterium-plate growth medium combinations and the result was recorded as a profile of growth inhibition zones. The sample profiles were compared to two sets of references: one constructed with standard antibiotic solution profiles, and the other with these combined with profiles of microbiologically and chemically identified residues from incurred samples. The algorithm employed in profile comparison located the minimal sum of absolute pairwise differences over the tests, with the addition of a number of experimentally observed intra-test criteria. Chemical identification and quantitation of incurred residues was based on liquid chromatography. The method identified penicillin G as a penicillinase sensitive penicillin, enrofloxacin and ciprofloxacin belonging to fluoroquinolone group, and oxytetracycline belonging to tetracycline group. Each of these residues was microbiologically identified below the Maximum Residue Limit (MRL) for kidney tissue. Combining sample profiles with the standard reference data set did not enhance the resolution. Microbiological and chemical identification test results were in good agreement. The results of this study show that a microbiological identification method is a useful tool in preliminary characterisation of antibiotic residues in animal tissues.

Incurred penicillin G, oxytetracycline, enrofloxacin and ciprofloxacin residues in bovine and porcine muscle and kidney samples were analysed by microbiological and chemical methods, the former using Bacillus subtilis BGA as a test organism on agar media of pH 6, pH 7.2 and pH 8 and the latter using liquid chromatography. Least squares fits between the logarithms of the chemically obtained concentrations of the antimicrobials and the widths of the inhibition zones were used to estimate the inhibition zone widths corresponding to the maximum residue limit concentrations. In vitro sensitivities were determined with standard antimicrobial solutions. The results indicate that if B. subtilis BGA is used as a test organism, muscle tissue cannot be used as test material for screening oxytetracycline, enrofloxacin and ciprofloxacin residues on the plates used in this study, while penicillin G can be screened from muscle tissue. Because of the numerous factors causing or increasing variation in the analysis, the inhibition zone caused by a given antibiotic concentration cannot be predicted precisely. Therefore, a positive agar diffusion test needs to be confirmed chemically. If a kidney sample gives a positive agar diffusion test result, the antimicrobial concentration in a muscle sample from the same carcass should be checked chemically.

The lead, cadmium and mercury concentrations in muscle, liver and kidney from Finnish pigs and cattle were determined. The average wet weight lead concentrations in pig muscle, liver and kidney were 15 micrograms/kg, 38 micrograms/kg and 40 micrograms/kg, respectively. The corresponding concentrations for cattle were 13 micrograms/kg, 57 micrograms/kg and 110 micrograms/kg. The average wet weight cadmium concentrations were 1.5 micrograms/kg, 28 micrograms/kg and 170 micrograms/kg (pigs) and 1.3 micrograms/kg, 61 micrograms/kg and 350 micrograms/kg (cattle). The corresponding mercury concentrations were 11 micrograms/kg, 12 micrograms/kg and 14 micrograms/kg (pigs) and 11 micrograms/kg, 12 micrograms/kg and 15 micrograms/kg (cattle). The average concentrations were at or above the detection limit of the metal in question. According to the results obtained by the National Veterinary Institute, the cadmium concentration in pigs and cattle has decreased during the period 1973-1988. The provisional tolerable daily intake of lead/person (60 kg), recommended by GEMS/Food, is 0.43 mg. According to the results for lead levels in these products in Finland, a daily intake of 29 kg pig muscle, 33 kg cattle muscle, 11 kg pig liver, 8 kg cattle liver, 11 kg pig kidney or 4 kg cattle kidney would be required to reach this norm. The corresponding provisional tolerable daily intake of cadmium/person (60 kg) is 0.06 mg and is equivalent to 40 kg pig muscle, 46 kg cattle muscle, 2 kg pig liver, 1 kg cattle liver, 0.4 kg pig kidney and 0.2 kg cattle kidney. The validity of the methods was tested four times a year using spiked check samples.

Microbiological and chemical identification of antimicrobial drug residues was attempted in 95 kidney and 76 muscle samples from 58 cattle, 36 pigs and one horse which had revealed kidneys positive to an inhibitor test. Information on pre-slaughter medication with one antimicrobial drug was available for 63% of the carcasses. Microbiological identification was performed by agar diffusion using 17 or 18 combinations of eight test bacteria, varying medium pH and three substances blocking the action of certain antimicrobials. Sample activity patterns compiled from inhibition zone diameters on test plates were compared with those obtained with standard antimicrobial solutions both visually and by locating the minimal sum of absolute pairwise differences over the tests. Chemical identification of residues was based on liquid chromatography. In kidney samples containing one microbiologically-identified antimicrobial the two methods gave fully consistent results with tetracyclines (15/15) and fluoroquinolenes (8/8). Preparation and storage of the kidney samples before chemical analyses appeared to influence the chemical identification of penicillin G. The results were consistent in 37 of the 41 samples stored without homogenization at -70 degrees C. The residue was identified by chemical means only in six and neither microbiologically nor chemically in four kidney samples with information on pre-slaughter medication. The same residue as in the kidney samples was identifiable microbiologically in 41% of the muscle samples of the same carcasses. The results show that the microbiological method is well suited for identification of antibiotic residues. They indicate further that an enhanced resolution with a reduced combination of plates is attainable.