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The effects of preservatives (potassium dichromate and sodium azide), heat treatment (untreated and 82 degrees C/10 min), and lactation stage upon the response of the microbial tests (BRT AiM and Delvotest) utilized for the detection of residues of antimicrobial substances in ewe milk were examined. Milk samples were collected from the morning milking of 50 Manchega ewes every 2 wk, from 15 d postpartum until the end of lactation. A total of 2322 samples were analyzed by BRT AiM with prediffusion and Delvotest microbial tests. The specificity of preservative-free milk samples without heat treatment was high (96.3% for BRT and 97.7% for Delvotest), with results improving for those samples thermally treated at 82 degrees C/10 min (99.0% for BRT and 98.7% for Delvotest). Potassium dichromate produced a total inhibition of growth of Bacillus stearothermophilus with both methods. When acidiol was utilized, the specificity of the samples not treated thermally was lower compared with preservative-free milk samples for the BRT AiM (90.2%) and Delvotest (91.0%) methods, improving when the samples were thermally treated, both for BRT AiM (94.8%) and Delvotest (95.3%), given that the presence of the preservative increased the frequency of doubtful results. The lactation stage significantly affected the results of the methods, with a greater frequency of false-positive and doubtful cases toward the end of the cycle, especially in those samples preserved with acidiol. The greater selectivity in both methods was therefore obtained for preservative-free ewe milk samples with prior heat treatment taken at the beginning or in midlactation period.

This experiment studied the effect of milking pipeline height (mid- vs. low-level milking system) on milking efficiency and milk composition. The experiment was of 8 wk duration: 2 wk preexperimental period and 6 wk experimental, in crossover design (2 x 2). Ewes were milked in a 2 x 12 milking parlor with 2 milking pipelines set at a milking vacuum of 36 kPa with a pulsation rate of 180 cycle/min and ratio of 50%. Height of the milkline had no effect on yield of milk at the time of milking, yield after stripping, milk composition, SCC, and number of teatcup fall-offs. Nor did milkline height have any effect on milk lipolysis or on the distribution of fatty acids. The level of free fatty acids was higher in evening than in morning milk (60.5 vs. 25.6 mg/L). Likewise, the increase in the degree of lipolysis between the receiver (40.4 mg/L) and the refrigeration tank (45.8 mg/L) underlines the importance of the milk delivery line design. The parameters (time and flow rate) that define the first peak in the milk emission kinetics were statistically different between lines, so care must be taken when comparing milk emission curves from both types of pipeline.

The presence of residues of antimicrobial substances in milk is one of the main concerns of the milk industry, as it poses a risk of toxicity to public health, and can seriously influence the technological properties of milk and dairy products. Moreover, the information available on the thermostability characteristics of these residues, particularly regarding the heat treatments used in control laboratories and the dairy industry, is very scarce. The aim of the study was, therefore, to analyze the effect of different heat treatments (40 degrees C for 10 min, 60 degrees C for 30 min, 83 degrees C for 10 min, 120 degrees C for 20 min, and 140 degrees C for 10 s) on milk samples fortified with three concentrations of nine beta-lactam antibiotics (penicillin G: 3, 6, and 12 microg/liter; ampicillin: 4, 8, and 16 microg/liter; amoxicillin: 4, 8, and 16 microg/liter; cloxacillin: 60, 120, and 240 microg/liter; cefoperazone: 55, 110, and 220 microg/liter; cefquinome: 100, 200, and 400 microg/liter; cefuroxime: 65, 130, and 260 microg/liter; cephalexin: 80, 160, and 220 microg/ liter; and cephalonium: 15, 30, and 60 microg/liter). The method used was a bioassay based on the inhibition of Geobacillus stearothermophilus var. calidolactis. The results showed that heating milk samples at 40 degrees C for 10 min hardly produced any heat inactivation at all, while the treatment at 83 degrees C for 10 min caused a 20% loss in penicillin G, 27% in cephalexin, and 35% in cefuroxime. Of the three dairy industry heat treatments studied in this work, low pasteurization (60 degrees C for 30 min) and treatment at 140 degrees C for 10 s only caused a small loss of antimicrobial activity, whereas classic sterilization (120 degrees C for 20 min) showed a high level of heat inactivation of over 65% for penicillins and 90% for cephalosporins.

The presence of drug residues in ewe's milk samples can be determined by microbial assays. The main limitation of these tests is the large number of false-positive results associated with them. False-positive results can be explained by the interaction of certain substances naturally existing in ewe's milk with the growth of the microorganism used in the test. In this study, milk chemical composition (fat, protein, lactose, total solids), somatic cell counts (SCCs), free fatty acid concentrations, and lactoperoxidase system components were determined in order to investigate their influence on the rate of false-positive results for the BRT and Delvotest microbiological inhibitor tests. Milk samples were obtained after morning milking of Manchega ewes at 15, 30, 45, 60, 75, 90, 105, 120, and 135 days after parturition. The animals did not receive any kind of treatment or medicated feed throughout the experiment. The false-positive rates for BRT and Delvotest were 3.75 and 2.4%, respectively. When the logistic regression model was applied, the percentages of total solids for positive samples were significantly different from those for negative samples (16.90 versus 18.42% for BRT, 16.05 versus 18.45% for Delvotest), while the SCC logarithmic transformation was significantly higher for the positive samples than for the negative samples (5.38 versus 5.11 log units for BRT, 5.32 versus 5.11 log units for Delvotest). Moreover, Delvotest-positive samples exhibited thiocyanate concentrations higher than those of Delvotest-negative samples (8.18 mg/liter versus 6.85 mg/liter). Further analyses are needed to confirm the possible presence of antimicrobial residues in this particular type of milk sample.

Two experiments were carried out to compare mechanical milking in mid-level (ML) and low-level (LL) milkline in goats. The first trial used 40 intramammary infection (IMI)-free goats that had been milked in ML during a pre-experimental period of 4±1 weeks post partum. These animals were divided into two groups (n=20), randomly assigning each group to ML or LL milking for a 17-week experimental period. During this period, several strategies were applied to increase teat exposure to pathogens in both experimental groups. The IMI rate was the same in both experimental groups (30% of goats), although the majority of new infections appeared earlier in ML (weeks 1-5) than in LL (weeks 7-16). Teat-end vacuum range (maximum minus minimum vacuum) was higher in ML than in LL, but no significant differences were found in the remaining variables [milk production and composition, somatic cell count (SCC), frequency of liner slips+teatcups fall off]. In the second experiment, in a crossover design (54 goats in fourth month of lactation; 2 treatments, ML and LL, in 2 experimental periods each lasting 1 week) it was observed that both the milk fractioning (reduced machine milk and increased machine stripping) and average machine milk flow worsened slightly in ML milking; in contrast, no differences were observed in total milking time or teat thickness changes after milking. It was concluded that the differences found between ML and LL are not sufficiently important to discourage breeders from using ML in goat milking.

To study the effect of time elapsed from the moment of taking samples on lactoperoxidase system components, we analyzed the activity of the lactoperoxidase enzyme and the concentrations of thiocyanate and hydrogen peroxide in 46 individual samples of Manchega ewe milk. Samples were maintained at a temperature of 4 degrees C until analysis, which took place at 6, 12, 24, and 48 h after extraction. Decreases were observed in lactoperoxidase activity when the analyses were performed at 48 h and in the thiocyanate and hydrogen peroxide concentrations at 12 h compared with those carried out earlier. Consequently, when the components of the lactoperoxidase system or its antibacterial activity are studied, the time elapsed since the sampling commenced must be taken into account. Similarly, the time elapsed is important when carrying out bacterial counts or residue screening by microbiological methods, during which the lactoperoxidase system may interfere. To study the component changes in the lactoperoxidase system during lactation, samples obtained 15, 30, 45, 60, 75, 90, 105, 120, and 135 d postpartum from 48 Manchega ewes were used. Average lactoperoxidase activity, thiocyanate, and hydrogen peroxide concentrations were 3.46 U/ml, 6.89 mg/L, and 0.39 mg/L, respectively, with significant variations throughout lactation. The thiocyanate and hydrogen peroxide levels at different lactation stages seemed to be insufficient to activate the lactoperoxidase system. Nevertheless, this could be achieved by adding 5 mg/L of thiocyanate and 8 mg/L of hydrogen peroxide at any time during lactation.

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