Short communication: effect of storage and preservation on total bacterial counts determined by automated flow cytometry in bulk tank goat milk.

This study was designed to evaluate the effects of different storage conditions on total bacterial count (TBC) determinations made in goat bulk tank milk using an automated flow cytometry method. The storage conditions tested were storage temperature (refrigeration at 4 and 10 degrees C or freezing at -20 degrees C), the use of a preservative (no preservative, NP; azidiol, AZ; or bronopol, BR), and the age of the milk samples for each analytical condition (storage times at 4 degrees C: from 0 h to 5 d for NP; and from 0 h to 22 d for AZ and BR; storage times at 10 degrees C: from 24 h to 2 d for NP and from 24 h to 22 for AZ and BR; storage times at -20 degrees C: from 24 h to 22 d for NP, AZ, and BR). Significant effects on individual bacterial count (IBC) variation were shown by the bulk tank milk sample, preservative, storage temperature, interaction preservative x storage temperature, and milk age within the interaction preservative x storage temperature. In preserved samples, the highest IBC were obtained for AZ and the lowest counts were obtained in samples preserved with BR. Because of the variation in IBC recorded in BR-preserved samples, we recommend that BR should not be used for TBC determinations using the automated flow cytometry method. The NP samples stored at 4 and 10 degrees C showed significantly higher IBC at 24 h postcollection, also invalidating these analytical conditions for TBC analyses. The practical implications of our findings are that goat milk samples preserved with AZ and stored at 10 or 4 degrees C are appropriate for TBC by the BactoScan flow cytometry method for up to 24 h and 11 d postcollection, respectively.

Short communication: Detection limits of non-beta-lactam antibiotics in goat's milk by microbiological residues screening tests.

This study compares the performance of 4 antimicrobial residue screening tests [brilliant black reduction test AiM (Analytik in Milch Produktions- und Vertriebs GmbH, München, Germany), Delvotest MCS (DSM Food Specialties, Delft, the Netherlands), Eclipse 100 test (ZEU-Inmunotec SL, Zaragoza, Spain), and Copan Milk Test (Copan Italia S.p.a., Brescia, Italy)] used to detect 20 antimicrobial agents (aminoglycosides, macrolides, tetracyclines, sulfonamides, and quinolones) in goat's milk, according to International Dairy Federation guidelines. Composite milk samples from 30 antibiotic-free goats were used to prepare spiked milk samples and 11,520 analytical determinations were carried out. According to a logistic regression model, agreement coefficients were greater than 98% for most of the antibiotics, with higher b values obtained for macrolides. Neither tetracyclines nor quinolones were detected at European Union maximum residue limits. Only the Copan Milk Test and the Delvotest MCS were able to detect 3 antimicrobials below their maximum residue limits (neomycin, tylosin, and sulfadimethoxine). Given that these tests are used in control programs for goat's milk, our results indicate their sensitivity would need to be improved to guarantee safety for consumers.

Detection limits of four antimicrobial residue screening tests for beta-lactams in goat's milk.

This study was conducted to compare the detection limits (DL) of several antibiotic residue screening tests with the maximum residue limits (MRL) authorized by the EU according to the guidance for the standardized evaluation of microbial inhibitor tests of the International Dairy Federation. Composite antibiotic-free milk samples from 30 primiparous Murciano-Granadina goats in good health condition were used to prepare test samples spiked with different concentrations of each antimicrobial. In total, 5,760 analytical determinations of 10 beta-lactam antibiotics (penicillin-G, ampicillin, amoxicillin, cloxacillin, oxacillin, dicloxacillin, cefadroxyl, cefalexin, cefoperazone, and cefuroxime) were performed using 4 antibiotic residue screening tests: the brilliant black reduction test BRT AiM (AiM-Analytik in Milch Produktions-und Vertriebs GmbH, München, Germany), Delvotest MCS (DSM Food Specialties, Delft, the Netherlands), Eclipse 100 (ZEU-Inmunotec SL, Zaragoza, Spain), and the Copan Milk Test (CMT; Copan Italia SpA, Brescia, Italy). For each method, we estimated the detection limits of the antimicrobial agents using a logistic regression model. Using the CMT and Delvotest on samples spiked with the 8 antibiotics for which MRL were available, DL were at or below the MRL. The BRT test provided DL at or below the MRL for all of the agents except cefalexin, whereas the Eclipse 100 method failed to detect 4 antibiotics (ampicillin, amoxicillin, cloxacillin, and cefoperazone) at MRL or below. Logistic regression-determined levels of agreement were highest for the CMT method (98.6 to 100%) and lowest for Eclipse 100 (66.3 to 100%). In general, agreement levels indicated good correlation between observed results and those predicted by logistic regression. The lowest b values (closely related to test sensitivity) were recorded for the cephalosporins (0.074 to 0.430) and highest for penicillin G, ampicillin, and amoxicillin (11.270 to 11.504). Delvotest and CMT best fulfilled IDF criteria for the ideal test for detecting antibiotic residues in milk.

Evaluation of the MilkoScan FT 6000 milk analyzer for determining the freezing point of goat's milk under different analytical conditions.

The aim of this research was to evaluate the Milko-Scan FT 6000 (Foss Electric, Hillerød, Denmark) for determining the freezing point (FP) of goat's milk under different analytical conditions. The FP was determined in duplicate in 1,800 milk aliquots obtained from 45 bulk tank milk samples from 10 Murciano-Granadina goat herds, using the MilkoScan method and a reference thermistor cryoscopy method (Advanced Instrument Inc., Norwood, MA). Five different preservation strategies--no preservative, preservation with azidiol (0.006 or 0.018 g of sodium azide/100 mL), and preservation with bronopol (0.020 or 0.040 g/100 mL)--were then used to preserve the milk. For each preservation strategy, 8 different amounts of water were added (0, 1, 2, 3, 4, 5, 6, or 7% total volume). The results obtained with each method under these 40 analytical conditions were examined by comparison of means, comparison of the standard deviations of repeatability (s(r) and its relative value s(r)%), and a regression analysis. Under most analytical conditions, the FP was recorded as lower by the MilkoScan method, with a mean difference of 1.5 m degrees C compared with the reference method. Both methods showed similar repeatabilities (the overall s(r)% was 0.22% for the MilkoScan method and 0.20% for the reference method). In comparisons of the 2 methods, the highest regression coefficients were obtained with aliquots containing >3% added water. The best regression coefficients (0.85 to 1.02) were obtained for milk samples preserved with bronopol at 0.020 g/100 mL. These results allow the MilkoScan method to be used with goat's milk for screening purposes. The factors of added water, preservative, analytical method, lactose concentration, and the effect of the bulk tank milk sample within each lactose group contributed significantly to the observed variation in FP. For practical purposes, either of the bronopol concentrations could be used when determining the FP of goat's milk with the methods tested. However, the increase in the concentration of sodium azide in the azidiol formula contributed to an important reduction in the FP recorded. Thus, the type and concentration of preservative should be taken into account when interpreting FP values.

Influence of storage and preservation on fossomatic cell count and composition of goat milk.

This study was designed to evaluate the effects of different test conditions on the somatic cell count (SCC) and composition of goat milk. To this end, 3600 tests were performed on 1800 aliquots taken from 40 goat milk samples using a combined instrument set-up based on flow cytometry for SCC and Fourier transform infrared analysis for fat, total protein, lactose, total solids, and freezing point determinations. The conditions tested were storage temperature (refrigeration and freezing), use of a preservative [no preservative (NP), azidiol (AZ), and bronopol (BR)], and age of the milk samples at each storage temperature (24 h to 42 d at refrigeration temperature and 21 to 105 d at freezing temperature). Significant effects on logSCC variation were shown by the storage temperature, the preservation treatment, the interaction of storage temperature x preservation treatment, and milk age within the interaction of storage temperature x preservative. Highest counts were recorded in the BR-preserved milk samples (logSCC = 5.877), and lowest counts were recorded in milk samples preserved using AZ (logSCC = 5.803). The use of frozen/thawed samples led to a significantly decreased logSCC for the treatments AZ and NP; the logSCC was not modified when BR-preserved frozen/thawed samples were analyzed. During storage, variations in the SCC observed for BR-preserved samples stored at refrigeration temperature for up to 25 d and at freezing temperature for all times tested were always < 10%. The preservation treatment was the main factor affecting the milk composition variables examined. Highest values of most variables were obtained in the BR-preserved samples, and the lowest values were obtained in the AZ-preserved samples. The freezing point was lower in the preserved samples than in the NP samples. The levels of milk constituents recorded in the BR-preserved samples were independent of both the storage temperature and age of milk sample. Our findings indicate that the freezing point of goat milk must be interpreted according to the preservative used.