A Rapid Microbiological Method for Enumeration of Pseudomonas fluorescens from Broiler Chicken Carcasses

1997 ◽  
Vol 60 (4) ◽  
pp. 385-390 ◽  
Author(s):  
SCOTT M. RUSSELL
Keyword(s):  
Pseudomonas Fluorescens ◽  
Broiler Chicken ◽  
Brain Heart Infusion ◽  
Brain Heart Infusion Broth ◽  
Animal Origin ◽  
Plate Count ◽  
Isolation And Identification ◽  
Plate Count Agar ◽  
Spoilage Bacteria ◽  
Time Required

A method to selectively enumerate Pseudomonas fluorescens from fresh chicken carcasses in less than 24 h using capacitance microbiology was developed. Capacitance assays were conducted on whole-carcass rinses at 25°C using brain heart infusion broth (BHI) containing 25 μg of Irgasan per ml to obtain a detection time. The capacitance samples were spread plated on plate count agar for isolation and identification. From plates with the highest dilution, from each carcass, 4 colonies were randomly selected and identified. Seven species of bacteria including Pseudomonas fluorescens were responsible for capacitance detection times. Various antibiotics and chemicals were added to basal media or brain heart infusion broth with Irgasan and were evaluated to select for the growth of P. fluorescens. BHI broth containing 4 μg of nitrofurantoin, 120 μg of carbenicillin, and 25 μg of Irgasan, all per ml, was found to be optimal and was termed Pseudomonas fluorescens selective additive (PSA) (patent pending). In a second study, 12 carcasses were collected in each of three replicate trials. For each trial, 2 carcasses were sampled immediately and 2 were sampled after storage at 3°C on days 3, 6, 9, 12, and 15. The BHI-PSA broth was found to be excellent for enumeration of P. fluorescens from broiler chicken carcass rinses in assays using capacitance microbiology at 25°C. The time required to enumerate P. fluorescens for all samples (day 0 to 15) was <22.4 h. This method is rapid and would be a useful tool for determining the number of spoilage bacteria on fresh chicken and thus may possibly be used to predict the potential shelf life of fresh chicken and other foods of animal origin.

Comparison of Media for Determining Temperature Abuse of Fresh Broiler Carcasses Using Impedance Microbiology

1995 ◽  
Vol 58 (10) ◽  
pp. 1124-1128 ◽  
Author(s):  
SCOTT M. RUSSELL ◽  
DANIEL L. FLETCHER ◽  
NELSON A. COX
Keyword(s):  
Broiler Chicken ◽  
Brain Heart Infusion ◽  
Plate Count ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
E Coli ◽  
Plate Count Agar ◽  
Processing Plants ◽  
Microbiological Techniques ◽  
The Ideal

Experiments were conducted to determine the ideal medium for detection of temperature abuse of fresh broiler chicken using impedance microbiological techniques. In three separate trials, 15 ready-to-cook broiler chicken carcasses were obtained from the chiller exit of three separate processing plants. Five carcasses were sampled immediately (day 0), 5 carcasses were sampled after temperature abusing at 25°C for 12 h and holding at 3°C for 6 days (temperature abused), and the remaining 5 carcasses were sampled after holding at 3°C for 7 days (day 7 controls). Whole-carcass rinses were diluted by placing 1 ml from each carcass into 9 ml of each of the following media: (1) brain heart infusion broth (BHI), (2) EC broth with 3% added dextrose (ECD), (3) CM medium with 2% added dextrose (CMD), (4) EC broth (EC), and (5) CM medium (CM). The diluted samples were assayed in duplicate at 43°C using impedance microbiological techniques. Once a detection time (DT) was recorded, one ml of the sample was immediately recovered from the module well, diluted to 10−6, 10−7, and 10−8, and spread plated onto plate count agar. Two colonies from each carcass on plates with the highest dilution were randomly selected and identified. Since both gram-positive and gram-negative genera of bacteria were isolated from BHI-cultured carcass rinses and were responsible for changing the impedance of the medium, DTs were variable. EC and ECD media were not suitable for conducting temperature-abuse determinations. Using CMD medium to select for the growth of gram-negative bacteria, specifically E. coli, temperature-abuse determinations were more accurate than using a general medium, such as BHI. CMD appears to be the most effective medium tested to conduct temperature abuse determinations using impedance microbiological techniques.

scholarly journals ATP-Bioluminescence as a method to evaluated microbiological quality of UHT milk

2014 ◽  
Vol 66 (6) ◽  
pp. 1909-1916 ◽  
Author(s):  
A.F. Cunha ◽  
A.D. Lage ◽  
M.M. Pereira e Araújo ◽  
C.F. Abreu ◽  
A.R. Tassinari ◽  
...  
Keyword(s):  
Microbiological Quality ◽  
Brain Heart Infusion ◽  
Plate Count ◽  
Culture Methods ◽  
Uht Milk ◽  
Milk Samples ◽  
Plate Count Agar ◽  
Atp Bioluminescence ◽  
Mesophilic Microorganism

New approaches are needed to quickly indicate possible contamination of UHT milk, among them the technique of ATP-Bioluminescence. Therefore, the aim of this study was to compare the results of culture methods with the results of ATP-Bioluminescence technique of 102 UHT whole milk samples incubated at 48, 72, and 168 hours. UHT milk samples were analyzed for the presence of mesophilic and psychrotrophic aerobic microorganisms using Plate Count Agar (PCA), Brain-Heart Infusion (BHI) media and PetrifilmTM Aerobic Count (AC) plates. The ATP-Bioluminescence technique was applied through the Microbial Luminescent Screening (MLS) system. Significant correlations were found between counts of aerobic mesophilic microorganisms on PCA, PetrifilmTM AC, BHI and results of ATP bioluminescence technique (P≤0.05). The ATP-Bioluminescence technique had higher correlation with counting method in PCA than BHI media. At lower pass/fail limits of Relative Light Units (60, 50, 45 and 40 RLU), the number of samples identified as positive increased and statistically agreed with aerobic mesophilic microorganism counts (P>0.05). For the dairy industry, the ATP-Bioluminescence technique may become an important tool that assists the official methods to quickly monitor the microbiological quality of UHT milk though this will likely require a threshold below 150 RLU.

Temperature Equilibration Times of Plate Count Agar and a Comparison of 50 Versus 45 C for Recovery of Raw-Milk Bacteria1

1975 ◽  
Vol 38 (6) ◽  
pp. 319-322 ◽  
Author(s):  
C. N. HUHTANEN ◽  
A. R. BRAZIS ◽  
W. L. ARLEDGE ◽  
C. B. DONNELLY ◽  
R. E. GINN ◽  
...  
Keyword(s):  
Raw Milk ◽  
Plate Count ◽  
Equilibration Time ◽  
Standard Methods ◽  
Standard Plate Count ◽  
Milk Samples ◽  
Plate Count Agar ◽  
Standard Plate ◽  
Time Required ◽  
Tempering Time

Sixty raw milk samples were plated using “Standard Methods” agar tempered to 45 or 50 ± 1 C. The standard plate count was significantly lower with the agar at 50 C. Tempering time (to 44–46 C) of a flask of agar in a water bath was about 5–10 min longer than that of a comparable flask of water. Time required to reach the desired temperature depended upon the volume of agar in the flasks, the number of flasks, and the volume of the water in the bath. Up to an hour of equilibration time may be necessary for newly autoclaved agar to reach the recommended temperature (44–46 C). Insufficient tempering time might cause an excessively high plating agar temperature which might cause a reduction in bacterial counts, especially of a heat sensitive psychrotrophic bacterium.

A Rapid Microbiological Method for Enumerating Escherichia colifrom Broiler Chicken Carcasses

1998 ◽  
Vol 61 (10) ◽  
pp. 1375-1377 ◽  
Author(s):  
ANGELA L. EDMISTON ◽  
SCOTT M. RUSSELL
Keyword(s):  
Rapid Method ◽  
Broiler Chicken ◽  
Bacterial Species ◽  
Plate Count ◽  
Processing Plant ◽  
Medium Temperature ◽  
E Coli ◽  
Plate Count Agar ◽  
Significant Linear Correlation ◽  
Inspection Service

Experiments were conducted to evaluate a rapid method for enumerating Escherichia coli from broiler chicken carcasses. In three separate trials, carcasses were obtained from a commercial processing plant, temperature abused at 37°C for 0, 3, 6, 9, or 12 h, and then rinsed. E. coli were enumerated from carcass rinses using Petrifilm E. coli count plates (PC) and by placing the rinse into double-strength colifiform medium supplemented with 2% dextrose (CMD). The CMD mixture was placed into a Bactometer module and conductance was measured at 44°C. Once a detection time (DT) was recorded, the sample was immediately recovered from the module well, diluted, and spread onto plate count agar. Colonies on plates at the highest dilution from each module well were randomly selected and identified. After 0, 3, 6, 9, and 12 h of temperature abuse, E. coli was the bacterial species identified 97, 92, 88, 87, and 61% of the time, respectively. These results indicate that the medium/temperature combination was excellent for enumerating E. coli from samples that contain mixed microflora using conductance. Significant linear correlations were observed between time of abuse (TA) and log10 PC (LPC) or DT (R2 = 0.86 and R2 = −0.90, respectively). A significant linear correlation was observed between LPC and DT (R2 = −0.92). This rapid method (1 to 7.6 h) for enumeration of E. coli on chicken should provide a way to determine E. coli levels before a product is shipped, and it should aid the poultry industry in meeting the E. coli testing requirement of the U.S. Department of Agriculture Food Safety and Inspection Service pathogen reduction regulation.

Effect of Organic Acids and Temperature on Survival of Shigella flexneri in Broth at pH 4†,‡

2002 ◽  
Vol 65 (9) ◽  
pp. 1417-1421 ◽  
Author(s):  
LAURA L. ZAIKA
Keyword(s):  
Organic Acids ◽  
Tartaric Acid ◽  
Storage Temperature ◽  
Shigella Flexneri ◽  
Brain Heart Infusion ◽  
Brain Heart Infusion Broth ◽  
Ion Concentration ◽  
Plate Count ◽  
Control Medium ◽  
Hydrogen Ion Concentration

The survival of bacterial pathogens in acidified foods depends not only on the hydrogen ion concentration, but also on the type of acid and the storage temperature. Shigella flexneri is a foodborne pathogen that is acid tolerant. The survival of S. flexneri 5348 in brain heart infusion broth supplemented with 0.04 M acetic, citric, lactic, malic, or tartaric acid and adjusted to pH 4 with HCl or NaOH was studied. The control medium was brain heart infusion broth adjusted to pH 4 with HCl. Stationary-phase cells were inoculated into media at initial populations of 6 to 7 log10 CFU/ml and incubated at 4, 19, 28, and 37°C. A two-phase linear inactivation model was applied to plate count data to derive lag times (tL) and slopes of the curves, from which D-values and time required for a 4-log10 decrease in population (T4D) were calculated. In all cases, survival increased with decreasing temperature. For each acid, tL, the D-value, and T4D increased with decreasing temperature. All acids inhibited S. flexneri to some extent but to differing degrees as follows: lactic acid, acetic acid > citric acid, malic acid, tartaric acid > HCl. The T4D values for the control medium and for media containing acetic, citric, lactic, malic, and tartaric acids were 64, 47, 50, 34, 58, and 52 h, respectively, at 37°C and 2,607, 1,498, 1,905, 1,346, 1,726, and 2,134 h, respectively, at 4°C. The results of this study indicate that organic acids may aid in the inactivation of Shigella. However, these data also suggest that foods stored at or below room temperature containing low levels (<1%) of acids could cause illness if contaminated with Shigella.

scholarly journals The ability of Aliarcobacter butzleri strains isolated from foods of animal origin in Costa Rica to form biofilm

2021 ◽  
Vol 10 (2) ◽  
Author(s):  
Marco Chaves ◽  
Daniel Vazquez-Valverde ◽  
Heriberto Fernandez-Jaramillo ◽  
Marìa Laura Arias-Echandi
Keyword(s):  
Costa Rica ◽  
Food Processing ◽  
Food Industry ◽  
Brain Heart Infusion ◽  
Brain Heart Infusion Broth ◽  
Animal Origin ◽  
Cross Contamination ◽  
Waterborne Pathogen ◽  
Adherence Ability

Aliarcobacter butzleri is a zoonotic emerging food and waterborne pathogen widely distributed in nature. It is present in food processing environments and can easily be spread through the food industry because of its ability to form biofilm. The aim of this work was to determine the ability of strains isolated in Costa Rica from different food matrixes of animal origin to form biofilm. Thirty-eight A. butzleri strains previously isolated and identified from animal origin products were analyzed using the method described by Stepmovic et al. (2000), in three culture broths, brain heart infusion broth, Boer broth and Houf broth. Results showed that 67% of poultry origin strains, 62.5% of meat origin strains and just 8% of milk origin strains showed ability to form biofilm. The findings of this study confirm the adherence ability of A. butzleri to form biofilm, a characteristic that can promote dispersion and cross contamination along food industry processing lines.

scholarly journals Designing and Evaluation of Skin Extract Agar for Isolation of Microflora from Raw Buffalo Hide

IIOAB Letters ◽  
2014 ◽  
Vol 3 (1) ◽  
Author(s):  
Ashish Polkade ◽  
Prafulla Shede ◽  
Pradhnya Kanekar ◽  
Prashant Dhakephalkar ◽  
Seema Sarnaik
Keyword(s):  
Nutrient Medium ◽  
Nutrient Agar ◽  
Plate Count ◽  
Rrna Gene ◽  
Skin Extract ◽  
Isolation And Identification ◽  
Standard Plate Count ◽  
Extract Agar ◽  
Plate Count Agar ◽  
Standard Plate

Abstract Present study was aimed to design nutrient medium most suitable for isolation and enumeration of microbial flora associated with raw buffalo hide. Skin extract agar (SEA) was designed and standardized on the basis of its chemical analysis. SEA and nutrient medium supplemented with skin extract was inoculated with buffalo hide wash. Total viable count as well as diversity of microbial colonies were enumerated on SEA as well as on nutrient agar and standard plate count agar both supplemented with skin extract (1% v/v). Bacterial strains forming diverse types of colonies on the media tested were identified on the basis of their 16S rRNA  gene sequences. The SEA was found to yield higher number of bacteria and to support growth of Acinetobacter, Exiguobacterium and Stenotrophomonas which otherwise difficult to selectively isolate from buffalo hide using nutrient agar and standard plate count agar. Diversity of microbial colonies formed on SEA was significantly higher than that observed on nutrient agar or standard plate count agar. Feasibility of utilizing SEA as a microbiological medium for isolation and identification of microflora from raw buffalo hide was successfully demonstrated. Use of skin extract medium can maximize recovery of taxonomically distinct bacteria from raw buffalo hide. This basic study, with proper manipulations could lead to development of product for enumeration and isolation of bacteria from buffalo hides especially cattle pathogens related to skin diseases.

Capacitance Microbiology as a Means of Determining the Quantity of Spoilage Bacteria on Fish Fillets

1998 ◽  
Vol 61 (7) ◽  
pp. 844-848 ◽  
Author(s):  
SCOTT M. RUSSELL
Keyword(s):  
Pseudomonas Fluorescens ◽  
Fish Species ◽  
Storage Time ◽  
Storage Period ◽  
Plate Count ◽  
Fresh Fish ◽  
Fish Flesh ◽  
Spoilage Bacteria ◽  
Red Grouper ◽  
Highly Correlated

An experiment was conducted to determine if a method for enumeration of Pseudomonas fluorescens in less than 11 h could be used to predict potential spoilage of fresh fish of four species. In each of three separate replications (Rep), five boneless fillets from each species of fish, including rainbow trout (RT), Atlantic salmon (AS), red grouper (RG), and tilapia (T) were obtained fresh from a retail outlet. For each species, six 25-g samples of fish flesh were asceptically removed from each fillet, placed into a polyethylene bag, and stored at 3°C for 0,1, 2, 3,4, or 5 days. After storage, samples were analyzed for psychrotrophic plate count (PPC), Pseudomonas fluorescens plate counts (PFPC), and P. fluorescens capacitance detection times (PDT) and subjectively evaluated for odor (ODOR). PPC gradually increased on all fish species as storage time increased. In most cases, PFPC decreased slightly and then progressively increased as storage time increased. In Reps 1 and 2, PDT decreased gradually (indicating an increase in bacteria); however, in Rep 3, PDT were erratic and difficult to interpret. Odor increased gradually throughout the storage period for all fish species. Linear correlations (R2 > 0.80) were observed between PPC and day of storage (DAY) for all fish species and Reps except for RT and RG in Rep 3. PFPC correlated (R2 > 0.70) to DAY for all fish except RT in Rep 3 and RG in Rep 2. PDT was negatively correlated to DAY for RT and T in Rep 1 and for all fish in Rep 2. Odor scores were highly correlated (R2 ≥ 0.84) to DAY for all fish tested. PPC and PDT were negatively correlated for RT in Reps 1 and 2, AS in Rep 2, and T in Reps 1 and 2. Because results can be obtained in < 12 h, the capacitance procedure with further refinement may provide an excellent alternative to conducting PPC as a means of predicting potential spoilage of fish such that the fillets of inferior quality (i.e., those that will spoil rapidly) may be sent to distribution outlets that are known to move fish products quickly and are able to sell the fish before it spoils.

Growth and Production of Enterotoxins A and D by Staphylococcus aureus in Salad Bar Ingredients and Clam Chowder

1991 ◽  
Vol 54 (11) ◽  
pp. 844-847 ◽  
Author(s):  
HASSAN GOURAMA ◽  
W. Y. J. TSAI ◽  
L. B. BULLERMAN
Keyword(s):  
Staphylococcus Aureus ◽  
Brain Heart Infusion ◽  
Brain Heart Infusion Broth ◽  
Plate Count ◽  
Green Pepper ◽  
Blue Cheese ◽  
Salad Dressing ◽  
Cell Counts ◽  
Elisa Technique ◽  
Number Of Cells

Growth and production of enterotoxins A and D (SEA, SED) by two strains of Staphylococcus aureus were determined in salad bar ingredients and clam chowder. Salad bar ingredients included lettuce, canned black olives, canned green olives, tomato, green pepper, blue cheese salad dressing, blue cheese crumbles, celery, and croutons. Total S. aureus were determined by plate count on Baird-Parker agar. Enterotoxins were quantified by using an ELISA technique. S. aureus did not survive in salad dressing, with pH 4.3. With the exception of olives and blue cheese, S. aureus survived on all ingredients for more than 12 h. After 24 h, the total number of cells decreased on most of the ingredients. S. aureus grew well on green pepper during the first 24 h, reaching 105 CFU/g, but no enterotoxins were detected. S. aureus also increased in moist and dry plain croutons, but there was no detectable production of enterotoxins. S. aureus growth was excellent in clam chowder with cell counts exceeding 108 CFU/g after 12 h at 42°C. Production of SEA and SED began shortly after 3 h. Maximal levels of SEA and SED were 0.29 and 1.6 ng/g, respectively, after 12 h. In brain heart infusion broth, the production of SEA and SED reached 21.9 and 36.3 ng/ml, respectively, after 24 h at 37°C.

scholarly journals Study on the isolation and identification of Pasteurella species from farm animal and human

2008 ◽  
Vol 2 (1) ◽  
pp. 81-90
Author(s):  
Sawsan salman Atia mubarak ◽  
Ismail khatham Shubar ◽  
Amar salim Rahiem
Keyword(s):  
Pasteurella Multocida ◽  
Agar Medium ◽  
Brain Heart Infusion ◽  
Farm Animal ◽  
Brain Heart Infusion Broth ◽  
Laboratory Animals ◽  
Isolation And Identification ◽  
Selective Agar

This study was interested in isolation and identification of Pasteuralla species in animals and human , also studing it's sensitivity to antibiotic with comparison to it's pathogenicity for laboratory animals . Tow groups of samples were collected and investigeted, 1st group consisting of one handered thirty six samples which were collected from animals ( cow .sheep , goats , and chikens ) . 2nd group was also consisting of 136 samples that were colleeted from human ( wound , urine , and sputum ) . These samples were cultured in selective enrichement brain heart infusion broth and then in the new Pasteurella multocida selective agar medium .

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