Effect of Agitation on Bacterial Aggregates in Pure Cultures and Raw Milk

1983 ◽  
Vol 46 (8) ◽  
pp. 681-685 ◽  
Author(s):  
ROBYN E. O'CONNOR ◽  
K. N. EWINGS ◽  
NEIL W. HOLLYWOOD
Keyword(s):  
Raw Milk ◽  
Mechanical Agitation ◽  
Filter Technique ◽  
Milk Samples ◽  
Pure Cultures ◽  
Plate Counts ◽  
Bacterial Aggregates ◽  
Direct Epifluorescent Filter Technique ◽  
Clump Size

A comparison of the effects of various mechanical agitation treatments on bacterial aggregates was performed on 8 pure cultures and 27 raw milk samples. Although both syringing and blending produced significant increases in total counts and psychrotroph counts, blending for 2 min gave the greatest increase in count. Use of the direct epifluorescent filter technique (DEFT) confirmed that syringing and blending reduced bacterial clump size to approximately 2 cells. These agitation treatments markedly improved the correlation between DEFT counts and plate counts.

Determination of Viable Bacterial Populations in Raw Milk within 20 Minutes by Using a Direct Epifluorescent Filter Technique

1997 ◽  
Vol 60 (7) ◽  
pp. 874-876 ◽  
Author(s):  
CLAUDE P. CHAMPAGNE ◽  
NANCY J. GARDNER ◽  
JULIE FONTAINE ◽  
JACQUES RICHARD
Keyword(s):  
Raw Milk ◽  
Plate Count ◽  
Bacterial Populations ◽  
Filter Technique ◽  
Milk Samples ◽  
Standard Plate ◽  
Plate Counts ◽  
Direct Epifluorescent Filter Technique ◽  
Triton X

The results from a shortened procedure for the direct epifluorescent filter technique (DEFT) determination of viable bacterial populations in raw milk were compared to standard plate counts. Shortening the prefiltration trypsin-Triton X-100 incubation period from 10 to 3 min enabled the completion of the analysis within 20 min. The short DEFT method results had a correlation coefficient (r) of 0.81 with plate counts. With respect to precision, the average difference between values of duplicate plate count analyses was 0.16 log units; that of the short DEFT was 0.14 log units. The slopes of the regressions equations were less than 1, indicating that a direct correlation is not achieved. Short DEFT values were 0.17 log units higher than those of plate counts on milk samples containing less than 10,000 CFU/ml. For milk samples containing counts over 10,000 CFU/ml, short DEFT values averaged only 0.05 log units above plate count readings. Daily preparation of the stain appears unnecessary since acridine orange solutions stored for up to 2 days at 4°C did not produce results significantly (P > 0.05) different from those obtained with fresh solutions. The short DEFT method has potential for the assessment of the bacteriological quality of raw milk in tanker deliveries.

BACTERIAL COUNTS OF RAW MILK AND FLAVOR OF THE MILK AFTER PASTEURIZATION AND STORAGE

1972 ◽  
Vol 35 (4) ◽  
pp. 203-206 ◽  
Author(s):  
G. B. Patel ◽  
G. Blankenagel
Keyword(s):  
Raw Milk ◽  
Bacterial Counts ◽  
Psychrotrophic Bacteria ◽  
Milk Samples ◽  
Standard Plate ◽  
Plate Counts ◽  
Subsequent Storage ◽  
And Storage ◽  
Common Defect

A total of 216 raw milk samples with a variety of Standard Plate Counts and psychrotrophic bacteria counts were laboratory-pasteurized, stored at 7 C, and then evaluated for flavor after 1 and 2 weeks. Results showed that milk with counts of >1,000,000/ml before heating frequently developed objectionable flavors after pasteurization and subsequent storage. The most common defect was a bitter flavor which appeared within 2 weeks after pasteurization in nearly all samples which as raw milk had counts exceeding 10,000,000/ml. This off-flavor developed in spite of small numbers of organisms in the pasteurized product and in the absence of post-pasteurization contamination.

Rapid Detection of Psychrotrophic Bacteria In Manufacturing Grade Raw Milks

1991 ◽  
Vol 54 (11) ◽  
pp. 861-867 ◽  
Author(s):  
S. R. TATINI ◽  
P. MEKALA ◽  
A. EL-HABAZ ◽  
M. W. GRIFFITHS
Keyword(s):  
Rapid Detection ◽  
Bacterial Count ◽  
Raw Milk ◽  
Plate Count ◽  
Psychrotrophic Bacteria ◽  
Milk Samples ◽  
Microscopic Count ◽  
Direct Epifluorescent Filter Technique ◽  
Native Microflora

Methods to rapidly assess the bacteriological quality of raw milk were investigated. Whereas direct microscopic count, modified psychrotrophic plate count, and direct epifluorescent filter technique (DEFT) did not correlate well with initial psychrotrophic bacterial count of raw milk, improvements were obtained after preincubation of the milk samples. The best preincubation conditions were identified as 30°C for 6 h, 21°C for 10 h, 13°C for 15 h, 13°C for 20 h, or 7°C for 37 h. The “square root” equation was applied to the data, and a model was produced for predicting growth of the native microflora of raw milk. Using this equation, a DEFT count after preincubation of the milk at 21°C for 10 h could accurately predict the initial psychrotroph count and the count after storage of the milk at 6°C for 48 h.

Effect of Five Surface Area-to-Sample Volume Ratios During Preliminary Incubation on The Bacterial Count of Raw Milk1,2

1979 ◽  
Vol 42 (12) ◽  
pp. 968-970
Author(s):  
J. A. RITTER ◽  
B. E. LANGLOIS ◽  
J. O'LEARY
Keyword(s):  
Surface Area ◽  
Incubation Temperature ◽  
Bacterial Count ◽  
Raw Milk ◽  
Sample Volume ◽  
Bacterial Counts ◽  
Milk Samples ◽  
Standard Plate ◽  
Plate Counts ◽  
Incubation Temperatures

Effects of ratio of surface area to volume of sample during preliminary incubation (PI) and of different plate incubation temperatures on bacterial counts of raw milk samples were studied. One hundred and twenty Grade A raw milk samples collected during a 10-month period were divided into five 100-ml aliquots and allotted to one of five surface area to volume ratios. The ratios during PI ranged from 0 cm2/100 ml to 149.74 cm2/100 ml. Following PI, pour plates of each treatment were incubated at 26, 30 and 32 C for 72 h. The Standard Plate Counts (SPC) ranged from 89 × 101 to 20 × 108/ml, with the SPC of 73.6% of the samples being less than 1 × 105/ml. Counts after PI tended to be higher as the plate incubation temperature decreased from 32 to 26 C and as the ratio of surface area to volume of sample increased. None of the differences between the counts for the 15 treatment-incubation temperatures was significant. Counts of 61 samples increased less than one log count during PI, while counts of 33 and 16 samples increased one to two log counts and over two log counts, respectively. The greater the SPC, the smaller the increase in count during PI. Of the 81 samples with SPC less than 1 × 105/ml, 29 had counts after PI that exceeded 2 × 105/ml.

The Automated Pyruvate Method as a Quality Test for Grade A Milk

1978 ◽  
Vol 41 (3) ◽  
pp. 168-177 ◽  
Author(s):  
R. T. MARSHALL ◽  
C. C. HARMON
Keyword(s):  
Incubation Temperature ◽  
Micrococcus Luteus ◽  
Raw Milk ◽  
Salmonella Typhi ◽  
Pseudomonas Fragi ◽  
Pasteurized Milk ◽  
Psychrotrophic Bacteria ◽  
Natural Flora ◽  
Pure Cultures ◽  
Plate Counts

Concentrations of pyruvate in Grade A raw and pasteurized milks were determined by an automated procedure. The method was sensitive to 0.1 mg of pyruvate per liter and the coefficient of variation of the method was 1.6% for a sample of milk containing 3 mg/1. Since about twice as much pyruvate was produced in samples incubated 24 h at 20 C compared with 15 C, the higher temperature is suggested for the pyruvate difference test (ΔP); however, psychrotrophs would more likely be detected with an incubation temperature of 15 C. Pyruvate difference tests of pure cultures in steamed milk distinguished between concentrations approximating 102 and 103 Pseudomonas fragi, 103 and 104 Pseudomonas fluorescens, and between 104 and 105 Escherichia coli, Streptococcus faecalis, Lactobacillus acidophilus and Micrococcus luteus, but did not distinguish between 104 and 105 initial concentrations of six other gram-positive bacteria and Salmonella typhi. Both the initial pyruvate (IP) concentration and the ΔP in raw milk were correlated with the logarithm of the Wisconsin Mastitis Test score, suggesting that somatic cells contribute to pyruvate content. Correlations of IP and ΔP with Standard Plate Counts of raw and pasteurized milks were low or insignificant and only with raw milk from normal quarters was there a significant correlation of IP and ΔP with initial Psychrotrophic Plate Counts (PPC). Rates of pyruvate production by pure cultures in steamed milk and by the mixed natural flora of pasteurized milk were consistent with rates of growth. However, certain psychrotrophic bacteria reduced the concentration of pyruvate to undetectable levels after producing 10 or more mg/1. Suggestions are made regarding applicability of the test in controlling keeping quality of pasteurized milk and in receiving of bulk raw milk.

Comparing Three Methods for Counting Raw-Milk Pour Plates1

1979 ◽  
Vol 42 (9) ◽  
pp. 729-731
Author(s):  
J. A. WESLEY ◽  
B. E. LANGLOIS ◽  
J. O'LEARY
Keyword(s):  
Raw Milk ◽  
Regression Equations ◽  
Standard Methods ◽  
Manual Count ◽  
Milk Samples ◽  
Plate Counts ◽  
Electronic Probe ◽  
The Difference ◽  
Incubation Temperatures ◽  
Colony Counter

Six hundred Grade A raw milk samples were plated on Standard Methods agar and the pour-plate counts compared using three methods. Counts of each sample were determined after incubation at 26, 30 and 32 C for 72 h with an automatic colony counter (ACC). Quebec colony counter and hand tally (HT) and Quebec colony counter and electronic probe with digital register (DT). No significant differences were found between the counts obtained with the HT and the DT for any of the three incubation temperatures. The ACC counts were lower (P < .01) than the manual counts at each temperature. The ACC counts were .39, .35 and .31 log counts lower than the manual count at 26, 30 and 32 C, respectively. The ACC counts at 72 h were higher (P < .01) than the ACC counts at 48 h for all temperatures. The difference was about .41 log counts. The two manual counts were totally correlated, while the correlation between the ACC counts and the manual count; was 97. Regression equations were formulated for predicting the 72 h HT count from either the 48 or 72 h ACC count.

AN EVALUATION OF SIMPLIFIED METHODS FOR DETERMINING VIABLE COUNTS OF RAW MILK

1960 ◽  
Vol 23 (9) ◽  
pp. 275-277 ◽  
Author(s):  
C. B. Donnelly ◽  
L. A. Black ◽  
K. H. Lewis
Keyword(s):  
Rapid Method ◽  
Raw Milk ◽  
Accurate Method ◽  
Bacterial Counts ◽  
Milk Samples ◽  
Standard Plate ◽  
Plate Counts ◽  
Oval Tube

When statistically analyzed, the results of comparative, examinations of raw milk samples revealed that bacterial counts by oval tube and microplate methods were essentially equivalent to standard plate counts. The oval tube provides an economical and accurate method for determining the count of producers' milk and may be used in the laboratory or on the platform. The microplate provides an equally accurate but more rapid method than either the oval tube or standard plate since counts be obtained in 20 hours.

scholarly journals Staphylococcus aureus of raw cow's milk

2021 ◽  
Vol 23 (102) ◽  
pp. 53-59
Author(s):  
M. D. Kukhtyn ◽  
Y. V. Horiuk ◽  
V. Z. Salata ◽  
V. T. Klymyk ◽  
N. M. Vorozhbit ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Raw Milk ◽  
Cow’S Milk ◽  
Cow's Milk ◽  
Milk Samples ◽  
Before And After ◽  
Pure Cultures ◽  
Almost All

The presence of Staphylococcus aureus in raw milk is considered a direct danger to consumers. Almost all types of dairy products normalize the amount of Staphylococcus aureus in 1 g (cm3) of product, due to their ability to produce thermostable enterotoxins. The aim of the study was to investigate the role of incision of teats of healthy and sick mastitis cows and milking equipment for irradiation of cow's milk with Staphylococcus aureus. Sanitation of milking equipment in farms was carried out with alkaline detergents based on sodium hypochlorite and acid based on phosphoric acid. Blood agar with 5 % sodium chloride was used to isolate staphylococci from washes and milk. Species identification of pure cultures of staphylococci was performed using RapID Staph Plus kits. At the same time before and after milking the skin of teats with Udder Wash and Udder Forte reduces the incidence of Staphylococcus aureus to 4.1 ± 0.3 % of cases. Quantitative determination of staphylococci, including golden on the skin of the teats revealed a decrease in the latter from 283.5 ± 21.2 to 27.6 ± 2.1 CFU/cm3 flush after milking. It has been established that the number of staphylococci, including golden, on the skin of teats increases sharply under subclinical or clinical ointment. Thus, it was found that in the subclinical form of staphylococcal mastitis from the skin of teats isolated S. aureus in the amount of 790.6 ± 65.4 CFU/cm3 of wash. At the same time, in the clinical form of mastitis, the amount of S. aureus increased, on average to 8 thousand CFU/cm3 of flush. It was found that under the condition of sanitary treatment of milking equipment with alternate use of alkaline and acid detergents and disinfectants, only 25 % of washes were staphylococci, and gold – up to 5 % of samples. It was found that in 43.7 ± 3.9 % of raw milk samples Staphylococcus aureus was not detected in 1 cm3. The number of milk samples containing Staphylococcus aureus up to 100 CFU/cm3 was 39.1 ± 3.3 %, and 17.2 ± 1.4 % of milk samples were contaminated with Staphylococcus aureus in the amount of 101 to 200 CFU/cm3. Therefore, the presence of more than 200 CFU/cm3 in freshly milked Staphylococcus aureus in milk indicates the ineffectiveness of antimastitis measures, skin remediation of teats and sanitation of milking equipment.

PRELIMINARY INCUBATION OF RAW MILK SAMPLES AS AN AID IN EVALUATING BACTERIOLOGICAL QUALITY1

1964 ◽  
Vol 27 (11) ◽  
pp. 333-338 ◽  
Author(s):  
M. N. Desai ◽  
T. J. Claydon
Keyword(s):  
Raw Milk ◽  
Growth Ratio ◽  
Milk Samples ◽  
Count Range ◽  
Before And After ◽  
Different Seasons ◽  
Standard Plate ◽  
Plate Counts ◽  
The Relationship ◽  
Summer Growth

A total of 194 milk samples were obtained from 35 bulk tank grade-A milk producers in the Manhattan, Kansas, milk-shed during different seasons, and examined for standard, coliform, and psychrophile plate counts both before and after preliminary incubation (P.I.). Samples also were tested for inhibitory substances. Farms were ranked I, II, or III in general sanitation on the basis of visible conditions, with rank I most desirable. Except for one doubtful sample, all tests for inhibitory substances were negative. With standard plate counts, when no consideration was given to initial count level, there was no statistically significant relation between P.I. counts and season or sanitation rank, although summer growth ratios were always lowest. When data were grouped on the basis of initial count range, P.I. counts and growth ratios increased with increased sanitation rank in the lower count ranges. In initial count ranges > 50,000/ml the relationship between growth ratio and sanitation rank declined. With coliform counts, considerable inconsistency occurred in P.I. counts among seasons and sanitation ranks. Initial and P.I. coliforms counts showed little relationship with standard plate counts (except in summer) or with psychrophile counts. With psychrophile counts, growth ratios during P.I. were influenced by season, being highest in winter and lowest in summer. P.I. counts generally increased with increase in sanitation rank. The relation between growth ratios and sanitation rank was statistically significant (P <0.05).

PREDICTION OF STANDARD PLATE COUNT OF MANUFACTURING-GRADE RAW MILK FROM THE PLATE LOOP COUNT1

1970 ◽  
Vol 33 (4) ◽  
pp. 168-170 ◽  
Author(s):  
Earl O. Wright ◽  
George W. Reinbold ◽  
Leon Burmeister ◽  
James Mellon
Keyword(s):  
Raw Milk ◽  
Plate Count ◽  
Adjustment Factor ◽  
Bacterial Counts ◽  
Standard Plate Count ◽  
Milk Samples ◽  
Count Range ◽  
Standard Plate ◽  
Plate Counts

Plate Loop Counts and Standard Plate Counts of manufacturing grade raw milk samples were compared. With bacterial counts of 200,000/ml and less, the Plate Loop Count approximated the Standard Plate Count very closely. With counts greater than 200,000, the agreement was not as close. Differences between counts from the two methods, however, were not statistically significant (P<0.05) within either count range. An adjustment factor was determined to enable the Plate Loop Count to more accurately predict the Standard Plate Count of milk with a Standard Plate Count >200,000/ml. The Standard Plate Count can be reasonably predicted from the Plate Loop Count by the formula:

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