Evaluation of a direct epifluorescent filter technique (DEFT) for the enumeration of bacterial spores in raw milk

1991 ◽  
Vol 1 (4) ◽  
pp. 253-261 ◽  
Author(s):  
Lynn Moran ◽  
M.T. Rowe ◽  
A. Gilmour
Keyword(s):  
Raw Milk ◽  
Bacterial Spores ◽  
Filter Technique ◽  
Direct Epifluorescent Filter Technique

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.

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.

scholarly journals Semi-Automated Direct Epifluorescent Filter Technique for Total Bacterial Count in Raw Milk

2000 ◽  
Vol 83 (6) ◽  
pp. 1345-1348 ◽  
Author(s):  
María Hermida ◽  
Margarita Taboada ◽  
Santiago Menéndez ◽  
José Luis Rodríguez-Otero
Keyword(s):  
Standard Deviation ◽  
Reference Method ◽  
Bacterial Count ◽  
Raw Milk ◽  
Total Bacterial Count ◽  
Filter Technique ◽  
Colony Forming Units ◽  
Repeatability Standard Deviation ◽  
Direct Epifluorescent Filter Technique ◽  
Different Levels

Abstract The Direct Epifluorescent Filter Technique (DEFT) and the reference method of counting total bacterial colonies on Petri dishes were compared. IDF Standards 128 (1985) and 161A (1995) were applied. A total of 496 samples of milk were analyzed. Colony forming units per μL milk were transformed to decimal logarithmic units: log (cfu/μL). The repeatability standard deviation, Sr = 0.114, was typical for a routine microbiological method. To study the carryover at different levels of bacteria, 3 tests were performed on milk samples of approximately 100, 700, and >1000 cfu/μL. For the first 2 experiments, no carryover was detected; in the milk sample with >1000 cfu/μL, the carryover was <0.12%. When the DEFT counts were regressed versus the reference method, the values of the slope and intercept were 0.92 and 0.17, respectively; the correlation coefficient was r = 0.84; and the residual standard deviation was Syx = 0.287. The paired t-test showed that the reference method and DEFT do not give significantly different results (p = 0.05).

The Limulus Amoebocyte Lysate Assay and the Direct Epifluorescent Filter Technique as Estimators of Potential Shelf-life of Pasteurized Fluid Milk

1990 ◽  
Vol 53 (2) ◽  
pp. 151-153 ◽  
Author(s):  
ROBERT D. BYRNE ◽  
J. RUSSELL BISHOP
Keyword(s):  
Shelf Life ◽  
Benzalkonium Chloride ◽  
Detection Techniques ◽  
Filter Technique ◽  
Psychrotrophic Bacteria ◽  
Limulus Amoebocyte Lysate ◽  
Whole Milk ◽  
Before And After ◽  
Direct Epifluorescent Filter Technique ◽  
Fluid Milk

The Limulus Amoebocyte Lysate Assay, Direct Epifluorescent Filter Technique and modified Psychrotrophic Bacteria Count were used to indicate potential shelf-life of pasteurized fluid milk. Commercial whole milk samples, stored at 7°C, were analyzed for bacterial and biochemical parameters, as well as for potential shelf-life by daily sensory evaluation. Each sample was evaluated before and after the following preliminary incubations: milk alone, milk with benzalkonium chloride, milk and broth, and milk and broth with benzalkonium chloride. The Limulus Amoebocyte Lysate Assay, Direct Epifluorescent Filter Technique, and modified Psychrotrophic Bacteria Count in conjunction with the preliminary incubations, produced relatively high correlations to shelf-life (−0.78, −0.85, and −0.86 respectively). Thus, these bacterial detection techniques could be used as rapid methods of shelf-life estimation.

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.

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