scholarly journals Rapid Methods for Detection and Enumeration of Campylobacter spp. in Foods

2002 ◽  
Vol 85 (4) ◽  
pp. 996-999 ◽  
Author(s):  
Haiyan Wang
Keyword(s):  
Dna Hybridization ◽  
Policy Development ◽  
Diarrheal Disease ◽  
Food Samples ◽  
Most Probable Number ◽  
Dot Blot ◽  
Probable Number ◽  
Rapid Methods ◽  
Cultural Methods ◽  
Campylobacter Spp

Abstract Campylobacter spp. are the most commonly reported bacterial cause of acute diarrheal disease in humans throughout the world. Traditional cultural methods for the detection and quantitation of Campylobacter spp. are slow and tedious; therefore, specific, sensitive, and rapid methods for campylobacters are needed to collect sufficient data for risk assessment and food safety policy development. We developed several rapid methods based on polymerase chain reaction (PCR), DNA hybridization, hydrophobic grid membrane filters (HGMFs), and enzyme immunoassays (EIAs). A PCR assay targeting C. jejuni, combined with a simple sample preparation procedure, detects as few as 0.3 most probable number (MPN)/mL C. jejuni in naturally contaminated chicken rinses after 20–24 h enrichment. An HGMF–EIA method using a commercial polyclonal antibody for Campylobacter detects and enumerates thermophilic Campylobacter spp. from spiked chicken rinse and milk, and naturally contaminated chicken rinses. A C. jejuni–specific probe in an HGMF–DNA hybridization protocol specifically detects and quantitates C. jejuni in food samples. A dot-blot EIA combined with an MPN procedure quantitates thermophilic campylobacters from samples that might be difficult to filter through HGMFs.

Development of Rapid Real-Time PCR and Most-Probable-Number Real-Time PCR Assays To Quantify Enterotoxigenic Strains of the Species in the Bacillus cereus Group

2007 ◽  
Vol 70 (12) ◽  
pp. 2774-2781 ◽  
Author(s):  
I-CHEN YANG ◽  
DANIEL YANG-CHIH SHIH ◽  
JAN-YI WANG ◽  
TZU-MING PAN
Keyword(s):  
Bacillus Cereus ◽  
Real Time ◽  
Real Time Pcr ◽  
Food Samples ◽  
Most Probable Number ◽  
Pcr Assay ◽  
Bacillus Cereus Group ◽  
Probable Number ◽  
Cooked Rice ◽  
Group Detection

Members of the Bacillus cereus group may produce diarrheal enterotoxins and could be potential hazards if they enter the food chain. Therefore, a method capable of detecting all the species in the B. cereus group rather than B. cereus alone is important. We selected nhe as the target and developed a real-time PCR assay to quantify enterotoxigenic strains of the B. cereus group. The real-time PCR assay was evaluated with 60 B. cereus group strains and 28 others. The assay was also used to construct calibration curves for different food matrices and feces. The assay has an excellent quantification capacity, as proved by its linearity (R2 > 0.993), wide dynamic quantification range (102 to 107 CFU/g for cooked rice and chicken, 103 to 107 CFU/ml for milk, and 104 to 107 CFU/g for feces), and adequate relative accuracy (85.5 to 101.1%). For the low-level contaminations, a most-probable-number real-time PCR assay was developed that could detect as low as 100 CFU/ml. Both assays were tested with real food samples and shown to be considerably appropriate for B. cereus group detection and quantification.

Occurrence and Characterization of Aeromonas Species in Pasteurized Milk and White Cheese in Rio De Janeiro, Brazil

1993 ◽  
Vol 56 (1) ◽  
pp. 62-65 ◽  
Author(s):  
ANGELA CÔRREA FREITAS ◽  
MARLY PAIVA NUNES ◽  
ARLETE MOREIRA MILHOMEM ◽  
ILVAN DELGADO RICCIARDI
Keyword(s):  
Rio De Janeiro ◽  
Foodborne Pathogen ◽  
Food Samples ◽  
High Rate ◽  
Most Probable Number ◽  
Aeromonas Species ◽  
Probable Number ◽  
Pasteurized Milk ◽  
Milk Samples ◽  
White Cheese

A total of 35 samples (1000 ml each) of pasteurized milk and 25 samples (100 g each) of white cheese purchased at supermarkets in Rio de Janeiro were analyzed for the presence of Aeromonas. Strains of Aeromonas were isolated from 28.5% of pasteurized milk and 32% of white cheese samples. Standard Plate counts in the pasteurized milk samples ranged from 7.2 × 10* to 2.5 × 105 CFU/ml. Total and fecal coliform counts in white cheese samples ranged from 1.9 × 10* to 2.4 × 105 most probable number per g and 3.2 × 102 to 1.2 × 105 most probable number per g, respectively. It was possible to identify Aeromonas caviae (58.9%), Aeromonas hydrophila (12.8%), and Aeromonas schubertii (2.5%) among the cultures isolated from pasteurized milk samples. Twenty-five percent of the strains could only be classified as Aeromonas spp. In white cheese samples, unclassified strains were the most frequent isolates (61.5%) followed by A. hydrophila (26.9%), A. caviae (7.6%) and Aeromonas sobria (3.8%). Only strains of A. hydrophila and A. sobria showed high rate of positive results when tested for the production of hemolysin, cytotoxin, and staphylolytic activity. Heat-stable enterotoxin and autoagglutination test did not correlate as virulence factors. The presence of Aeromonas species in refrigerated food samples suggests that this microorganism could be a potential foodborne pathogen, and dairy products may represent an important vehicle of its transmission.

Differentiation of Clostridium perfringens from Related Clostridia in Iron Milk Medium

1985 ◽  
Vol 48 (2) ◽  
pp. 130-134 ◽  
Author(s):  
CARLOS ABEYTA ◽  
ANITA MICHALOVSKIS ◽  
MARLEEN M. WEKELL
Keyword(s):  
Clostridium Perfringens ◽  
High Selectivity ◽  
Positive Response ◽  
Food Samples ◽  
Gas Production ◽  
The Other ◽  
Plate Count ◽  
Most Probable Number ◽  
Probable Number ◽  
Positive Results

The stormy fermentation reaction of Clostridium perfringens in iron milk medium was compared to that of several C. perfringens-like strains. These clostridia, C. barati, C. perenne, C. absonum, and C. paraperfringens are very similar to C. perfringens on the basis of certain biochemical reactions and, consequently, are often difficult to distinguish from C. perfringens. Furthermore, these related clostridia may also be present in foods. Results of this study demonstrate that after 18 h of incubation at 45°C, only C. perfringens gave a positive reaction in iron milk with inocula as low as 22 cells/g. Some of the other strains began to show only gas production at 18 h. After 24 to 42 h some strains gave positive results and after 72 h all were positive. Enumeration of C. perfringens from food samples in iron milk medium by a 3-tube most probable number (MPN) technique gave similar results to enumeration by plate count using Shahidi-Ferguson Perfringens (SFP) agar. Furthermore, a rapid positive response occurred after only 2 and 3 h incubation of iron milk inoculated with 108 and 107 cells/ml, respectively. The high selectivity, ease of identification and rapid growth of C. perfringens in iron milk make the iron milk MPN procedure a valuable assay for accurate enumeration and differentiation of C. perfringens from related Clostridia in food products.

Glucuronidase Assay in a Rapid MPN Determination for Recovery of Escherichia coli from Selected Foods

1987 ◽  
Vol 70 (1) ◽  
pp. 31-34
Author(s):  
Wallace H Andrews ◽  
Clyde R Wilson ◽  
Paul L Poelma
Keyword(s):  
Escherichia Coli ◽  
False Positive ◽  
Uv Light ◽  
Most Probable Number ◽  
Probable Number ◽  
Rapid Methods ◽  
Glucuronidase Assay

Abstract Glucuronidase is present in most strains of Escherichia coli but absent in most other enteric microorganisms; therefore, an assay for this enzyme is useful for determining the presence of the organism. The substrate 4-methylumbelliferyl beta-D-glucuronide (MUG) is incorporated into either lauryl tryptose (LT) broth or EC medium; the inoculated tubes are then incubated under specified conditions and examined under longwave UV light for the presence of a fluorogenic glucuronidase end product. When compared with the 10-day most probable number (MPN) procedure of AOAC, the LT-MUG and the EC-MUG tests required 24 and 96 h, respectively, and gave comparable mean log MPN values for samples of crabmeat, sunflower kernels, and walnut pieces. However, false-positive and falsenegative reactions were observed with foods tested by both of these rapid methods. Overall, method sensitivity was not compromised by using the LT-MUG rather than the EC-MUG method. Incorporation of 25 αg MUG/mL into LT broth resulted in diminished fluorescence of positive reactions, whereas MUG concentrations of 50 and 100 αg/mL provided decisive fluorogenic reactions.

Comparison of the SimPlate Coliform and Escherichia coli Test with Petrifilm, Three-Tube MPN, and VRBA + MUG Methods for Enumerating Coliforms and E. coli in Food

1998 ◽  
Vol 61 (4) ◽  
pp. 444-449 ◽  
Author(s):  
D. E. TOWNSEND ◽  
R. L. IRVING ◽  
A. NAQUI
Keyword(s):  
Escherichia Coli ◽  
Correlation Coefficients ◽  
Raw Milk ◽  
Food Samples ◽  
Coliform Bacteria ◽  
Most Probable Number ◽  
Additional Food ◽  
Suitable Alternative ◽  
Probable Number ◽  
E Coli

SimPlate for coliforms and Escherichia coli (CEc) is a new method for the detection and quantification of coliforms and E. coli in food. Internal validation of the method was carried out at IDEXX Laboratories (Westbrook, ME) with 180 food samples representing a variety of different food matrices and compared against three-tube MPN (most probable number), VRBA (violet red bile agar) + MUG, and Petrifilm (E. coli count) methods. SimPlate CEc was highly correlated with each of these methods for the quantification of coliform bacteria (r ≥ 0.90). An insignificant number of food samples were found to contain E. coli; therefore, no meaningful correlation data could be generated. Four hundred forty-four additional food samples were tested at five collaborating laboratories for the presence of coliforms and E. coli using SimPlate CEc and either VRBA + MUG or Petrifilm (E. coli count). Regression analysis of data from SimPlate for CEc versus Petrifilm E. coli count plates generated correlation coefficients (r) of at least 0.89 for total coliforms and at least 0.90 for generic E. coli. Correlation coefficients between SimPlate for CEc and VRBA + MUG data were at least 0.90 for coliforms and at least 0.86 for E. coli. SimPlate for CEc demonstrated better recovery of E. coli than Petrifilm when high populations of bacteria were present. E. coli was not detected in 20 of 50 (40%) raw milk samples tested by the Petrifilm method due to the presence of interfering coliform and noncoliform bacteria. It is concluded that SimPlate for CEc is a suitable alternative for determining numbers of coliform bacteria and E. coli in food.

Enumerative Analysis of Salmonella in Outbreak-Associated Breaded and Frozen Comminuted Raw Chicken Products

2017 ◽  
Vol 80 (5) ◽  
pp. 814-818 ◽  
Author(s):  
Angela Catford ◽  
Kyle Ganz ◽  
Sandeep Tamber
Keyword(s):  
Foodborne Pathogen ◽  
Food Samples ◽  
Most Probable Number ◽  
Distribution Analysis ◽  
Probable Number ◽  
Contaminated Food ◽  
Foodborne Outbreaks ◽  
Low Levels ◽  
Data Gap ◽  
Pathogen Exposure

ABSTRACT A significant data gap exists with respect to the levels of pathogens in foods implicated in foodborne outbreaks. These data are essential for the quantification of pathogen exposure via the ingestion of contaminated food. Here we report the levels of the foodborne pathogen Salmonella in comminuted raw chicken products that had been breaded and then frozen. The products investigated were collected during four food safety investigations of foodborne outbreaks that occurred in Canada from 2014 to 2016. Most-probable-number (MPN) distribution analysis of the food samples revealed Salmonella levels of 0.0018 to 3 MPN/g, which is equivalent to 1 MPN per 0.33 to 556 g of product. These data suggest low levels of Salmonella may be associated with foodborne outbreaks.

Fungi in Foods

1975 ◽  
Vol 38 (12) ◽  
pp. 745-746 ◽  
Author(s):  
J. A. KOBURGER ◽  
A. R. NORDEN
Keyword(s):  
Food Samples ◽  
Plate Count ◽  
Most Probable Number ◽  
Probable Number ◽  
Plate Count Agar ◽  
Yeasts And Molds ◽  
Surface Plate

It was possible to compare recovery of yeasts and molds from 30 food samples by three methods, employing plate count agar and broth with added antibiotics. Although the pour plate and surface plate methods gave comparable results, the Most Probable Number (MPN) procedure consistently yielded the highest counts. With some of the samples, the MPN method was the only one in which recovery occurred. It thus appears that this procedure is practical for detection of fungi and may be of use in survey work or when analyzing foods containing low numbers of microorganisms.

Comparison of the SimPlate™ Total Plate Count Method with Petrifilm™, Redigel™, and Conventional Pour-Plate Methods for Enumerating Aerobic Microorganisms in Foods

1998 ◽  
Vol 61 (1) ◽  
pp. 14-18 ◽  
Author(s):  
L. R. BEUCHAT ◽  
F. COPELAND ◽  
M. S. CURIALE ◽  
T. DANISAVICH ◽  
V. GANGAR ◽  
...  
Keyword(s):  
Food Samples ◽  
Test Methods ◽  
Plate Count ◽  
Most Probable Number ◽  
Suitable Alternative ◽  
Probable Number ◽  
Plate Count Agar ◽  
Total Plate Count ◽  
Wide Range ◽  
Highly Correlated

The SimPlate™ Total Plate Count (TPC) method, developed by IDEXX Laboratories, Inc., is designed to determine the most probable number of aerobic microorganisms in foods. The 24-h test was compared to the conventional plate count agar (PCA) method, the Petrifilm™ Aerobic Count plates, and the Redigel™ Total Count procedure for enumerating microflora in 751 food samples. Results using the SimPlate™ TPC method were highly correlated (r ≥ 0.96) with results from other test methods. Slopes (0.96–0.97) were not significantly different from 1, and y intercepts (−0.03–0.08) were not different from 0. The SimPlate™ has a high counting range (> 1600 most probable number per single dilution), thus requiring fewer dilutions of samples compared to other methods evaluated. Some foods, e.g., raw liver, wheat flour, and nuts, contain enzymes that gave false-positive reactions on SimPlates™. Overall, however, the SimPlate™ TPC method is a suitable alternative to conventional PCA, Petrifilm™, and Redigel™ methods for estimating populations of mesophilic aerobic microorganisms in a wide range of foods.

Evaluation of Cultural Methods to Isolate Salmonella from Pressed Yeast and Dried Inactive Yeast

1975 ◽  
Vol 38 (7) ◽  
pp. 383-385 ◽  
Author(s):  
CLYDE R. WILSON ◽  
WALLACE H. ANDREWS ◽  
PAUL L. POELMA
Keyword(s):  
Most Probable Number ◽  
Distilled Water ◽  
Lauryl Sulfate ◽  
Improved Method ◽  
Probable Number ◽  
Examination Procedure ◽  
Cultural Methods ◽  
Modified Method ◽  
Current Procedure ◽  
Better Than

An improved method recently developed to isolate Salmonella from dried active yeast was evaluated for use with pressed yeast and dried inactive yeast. The method for dried active yeast consists of pre-enriching a 25-g sample in trypticase soy (TS) broth at a sample-broth ratio of 1:10, incubating at 35 ± 0.5 C for 24 ± 2 h, and transferring to lauryl sulfate tryptose (LST) and tetrathionate (TT) broths. After 24 ± 2 h at 35 C, the broths are streaked to selective agars. When evaluated for use with pressed yeast, Salmonella attained higher most probable number levels/ml with this method (7.9 × 104 – 3.3 × 106), than with 1% tryptone broth and a sample-broth ratio of 1:5 (1.7 × 103 – 1.7 × 106), which is the current examination procedure for pressed yeast. Salmonella was consistently isolated from selective agars streaked from TT broth, but was seldom isolated from selective agars streaked from LST broth because of massive overgrowth by non-salmonellae. With dried inactive yeast, this modified method was equal to, but not significantly better than, pre-enrichment of the yeast in sterile distilled water at a sample-broth ratio of 1:5, which is the current procedure for dried inactive yeast.

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