HM Medical Clinic

 

Novamed.co.il

28 PIERRE KÖENIG ST., TALPIOT INDUSTRIAL AREA POB 53231 JERUSALEM 91531 ISRAEL TEL. 972-2-6781861 FAX. 972-2-6781852
e-mail: [email protected]
NOVASTREAK Microbial Contamination Monitoring Device
POSITIONING & STRATEGY
Traditional Culturing Methods (TCM) for the detection of pathogenic bacteria from foods rely on culturing the bacteria onto agar plates. TCM employs a streaking dilution of the sample on a solid agar media with a calibrated loop, providing accurate identification and quantification of individual colonies. These TCM are laborious and consuming, taking three days or more to detect specific pathogens. Nevertheless, one major disadvantage of this approach is that the samples must be transported to the microbiology lab, where skil ed personnel then plate it. Since food is an excel ent medium for bacterial culture, the number of bacteria present may increase rapidly if during transportation the sample is not refrigerated, which, of course, increases the cost of every test performed. Recent advances in technology make detection and identification faster, safer and more convenient than traditional assays. NOVASTREAK Microbial Contamination Monitoring Device (MCMD) is your superior device, designed to be one step ahead of al culturing devices and to further reduce hands-on involvement. The unique dip-tip technique dif erentiates NOVASTREAK MCMD from al other commercial dipslides enabling dilution streaking similar to culture. NOVASTREAK MCMD combines the sensitivity & specificity of the TCM, with the convenience of dipslide, which together with special unique features makes it the leading device for microbial contamination testing! The simple and rapid protocol together with the see-through view results in a unique transport packaging position NOVASTREAK MCMD as a premium high quality price product, providing easy to use friendly standardized culture process from the sampling site up to the microbiological laboratory.
INTENDED USE
NOVASTREAK MCMD is a convenient semi-quantitative screening culture device for sampling and assessing microbial contamination of food and dairy products, industrial fluids and surfaces of sanitary importance. A unique streaking mechanism permits the isolation of single colonies even when the original bacterial population of the sample was as high as 107 organisms per mil iliter. NOVASTREAK MCMD is intended for use in the food
SUMMARY AND EXPLANATION
NOVASTREAK MCMD comprises a plastic paddle with two types of agar at ached back-to-back, housed in a closed transparent plastic tube. A ring with elongated prongs is at ached to the end of the paddle so that there are prongs on each side of the slide. The ends of the prongs are dipped into the liquid sample. Upon re- insertion into the plastic tube, the prongs are prevented from moving and the agar surfaces are inoculated with the sample as the paddle passes over the prongs. The result is a series of streaks of decreasing bacterial concentration, which permits isolation of single colonies even when the original bacterial population of the sample was as high as 107 organisms per mil iliter. NOVASTREAK MCMD can be used to monitor microbial growth wherever the potential may exceed 103 microorganisms in ml of sample. NOVASTREAK MCMD provide a simple, faster, safer and convenient alternative to the TCM for detection of microbial present. NOVASTREAK MCMD unit consists of two dif erent agar modifications at ached back-to-back on a plastic sampling paddle, which is permanently fastened to the cap for comfort of handling during use. NOVASTREAK MCMD are self- contained units, which can be taken to the sampling site instead of transporting samples, back to the laboratory. NOVASTREAK MCMD is available with a variety of growth specific media and color indicators.
ADVANTAGES OF METHOD
The total extra-cost for TCM include: (1) expensive skil ed personnel for inoculation, (2) transportation to the laboratory, (3) special conditions (refrigeration) for transported specimens (4) time consuming preparing the solid agar in the lab by a technicians. NOVASTREAK MCMD is a leading product in comparison to the TCM with the fol owing advantages: (1) easy protocol – simple, rapid and handy, (2) no skil ed personnel are required due to the simple to use straightforward device, (3) sampling at the site of col ection – no special Lab facilities are required for preparation, (4) saves you a lot of space in the incubator due to its compact design and comfortable incubation stand.

SAMPLING
SAMPLE TYPE
MATERIAL TO BE TESTED
PROCEDURE
Milk (raw and pasteurized) Dip sampling procedure Compare with Colony Density Liquid samples
Industrial water (waste, recycled, cooling or process water), dairy products (starter cultures, sour cream, General streaking procedure Compare with Colony Density yogurt and other fermented products) Syrups, pasts and dehydrated Viscous and
products (vegetables, fruit, egg Dilute 1:1 or 1:10 in sterile Compare with Colony Density friable samples
powder, milk powder, powdered water with fol owing General Chart No. 1 and multiply the soups, instant desserts, cocoa, etc.) streaking procedure result by 2 or 10 Raw material, frozen and Homogenize and suspended Compare with Colony Density Solid samples
chil ed products ( meat, fish and sea 1:1 or 1:10 in sterile water with Chart No. 1 and multiply the fol owing General streaking result by 2 or 10 Surfaces
Utensils, work surfaces Touch surface with paddle for Compare with Colony Density
GENERAL PROCEDURE
A. STREAKING SAMPLING
1. Unscrew the NOVASTREAK MCMD cap. Pul the paddle out. Do not touch any part but the cap.
2. Hold the paddle vertical y and dip the white prongs into the sample up to about half of their length
3. Return the paddle to its container in a quick, continuous and vertical motion and tighten cap.
4. Transport the tube to laboratory for incubation and examination
5. Before incubation, loosen cap one-half turn.
6. Incubate the entire container at (35oC±2oC) for 18-24 hours in a vertical position.
7. Interpret the results by simple visual comparison of bacterial growth on the agar surface with
Colony Density Chart No. 1 provided. No actual colony counting is necessary.

B. DIP SAMPLING (DIPSLIDE TECHNIQUE)
1. Unscrew the NOVASTREAK MCMD cap. Pul the paddle out. Do not touch any part but the cap.
2. Immediately return the paddle to the tube (in order to move the prongs out of the way) and then pul
the paddle out again. 3. Dip the culture paddle into a diluted/undiluted sample or pour the sample over agar surfaces, if the
volume of sample is not adequate to ful y immerse the agar surfaces. 4. Replace inoculated culture paddle in its protective NOVASTREAK MCMD vial and close cap.
5. Transport NOVASTREAK MCMD vial to laboratory for incubation and examination.
6. Place inoculated NOVASTREAK MCMD vial upright in incubator (35oC±2oC) for 18-24 hours.
Before incubation, loosen cap one-half turn. 7. Interpret the results by simple visual comparison of bacterial growth on the agar surface with
Colony Density Chart No. 2 provided. No actual colony counting is necessary.

C. SURFACE CONTACT SAMPLING
1. Unscrew the NOVASTREAK MCMD cap. Pul the paddle out. Do not touch any part but the cap.
2. Immediately return the paddle to the tube (in order to move the prongs out of the way) and then pul
the paddle out again. 3. Gently touch the agar faces onto the surface to be tested. The agar should remain in contact for
about 20 seconds. 4. Replace inoculated culture paddle in its protective NOVASTREAK MCMD vial and close cap.
5. Transport NOVASTREAK MCMD vial to laboratory for incubation and examination.
6. Place inoculated NOVASTREAK MCMD vial upright in incubator (35oC±2oC) for 18-24 hours.
Before incubation, loosen cap one-half turn. 7. Interpret the results by simple visual comparison of bacterial growth on the agar surface with
Colony Density Chart No. 3 provided. No actual colony counting is necessary.
MATERIALS PROVIDED
REAGENTS
EXPECTED TYPES OF
REAGENTS
Side 1: Plate Count Agar (PCA)
Agar Color: Yel ow Total Bacterial Count Side 2: Oxytetracycline-Glucose-Yeast Extract
Agar Color: Yel ow Side 1: Violet Red Bile Agar (VRBA)
BD-506 Side 2: Ox tetracycline-Glucose-Yeast Extract Agar Color: Purple
Total Coliforms Count Agar Color: Yel ow BD-507 Side 1: Violet Red Bile Agar (VRBA)
Agar Color: Purple Total Coliforms Count Side 2: Plate Count Agar (PCA)
Agar Color: Yel ow Total Bacterial Count Agar Color: Yel ow, BD-508 Side 1: Baird Parker Agar
Side 2: Violet Red Bile Agar (VRBA)
Agar Color: Purple Total Coliforms Count Side 1: Oxytetracycline-Glucose-Yeast- Agar Color: Yel ow
BD-511 Extract Agar (OGYE)
Side 2: Rose Bengal Agar w.Chloramphenicol Agar Color: Pink
BD-521 Side 1: XLD Agar
Agar Color: Orange Salmonel a, Shigel a Side 2: Salmonel a ID Agar
Agar Color: Yel ow Salmonel a
APPLICATION FIELDS
TYPE OF MICROORGANISMS
MATERIAL TO BE TESTED
Total Coliforms Total Bacterial
Water (waste, recycled, cooling or process
Dairy Products (pasteurized milk, starter
cultures, sour cream, yogurts and other fermented dairy products) Meat, fish, sea food (raw material, frozen and
chil ed products) Surfaces
Syrups, pasts and dehydrated products
(vegetables, fruit, egg powder, milk powder, powdered soups, instant desserts, cocoa, etc.) MATERIAL REQUIRED BUT NOT PROVIDED
Incubator (37 ± 2°C) Incubation Stand WARNING AND PRECAUTIONS
1. For In Vitro Diagnostic Use.
2. Use aseptic technique and established laboratory procedure in handling and disposing of infectious
1. Store NOVASTREAK MCMD at 2-8oC up to 6 months, refer to product label.
2. Protect contents from direct light to ensure product stability through the expiration date, shown on
the tube cap or packaging label
KIT CONTENTS
REAGENTS
EXPECTED RESULTS
Plate Count Agar (PCA) is also known as Standard Methods
Agar. This medium contains casein hydrolyzate, dextrose and yeast extract. It is recommended for the isolation and A variety of bacterial and fungal species appearing as enumeration of bacterial and fungal microorganisms of milk contaminants in dairy and other food products wil show and other dairy products, and may be used to determine the good growth, including lactobacil i and staphylococci. sanitary quality of foods, water and other materials. Oxytetracycline-Glucose-Yeast-Extract Agar (OGYE) is
recommended for the selection and enumeration of yeasts and molds from foodstuf s. The medium uses oxytetracycline as OGYE agar, after 5 days at 22 + 3 o C produces good to the selective agent at a neutral pH, that gives increased counts excel ent growth of Aspergil us niger, Saccharomyces of yeasts and molds compared with media, which rely on a low cerevisiae, with total inhibition of E. coli. pH to suppress bacterial growth. Violet Red Bile Agar (VRBA) is a selective medium for the
detection of coliform organisms in water, milk, and other Lactose fermenters are rose-red in color and general y materials of sanitary importance. The medium is selective due surrounded by a halo of precipitated bile. E. coli colonies to the presence of inhibitors, bile salts and crystal violet. are entire edged, 1mm or more in diameter. E. Dif erentiation of enteric microorganisms is achieved by the aerogenes are larger, often mucoid and pinkish. Lactose combination of lactose and the neutral red indicator. Colorless non-fermenters produce colorless colonies. Enterococci or pink-to-red colonies are produced depending on the ability occasional y grow to produce rose colonies pinpoint in to ferment lactose. Baird Parker Agar is used for the selective isolation and
enumeration of coagulase-positive staphylococci from food, Typical colonies of S. aureus are black, shiny, convex skin, soil, air and other materials. It may also be used to and surrounded by clear halos of approximately 2 to 5 identify staphylococci on the basis of their ability to clear egg mm. Coagulase-negative staphylococci do not grow yolk (lecithinase production). Sodium pyruvate is incorporated wel ; if at al , the typical clear halos are absent. Other to impart S. aureus their black colony appearance. Glycine organisms are inhibited or may grow sparsely, producing and lithium chloride have inhibitory action on organisms other white to brown colonies with no clearing of the egg yolk. than S. aureus. Xylose Lysine Decarboxylase (XLD) Agar is recommended
for the isolation and dif erentiation of enteric pathogens, E. coli and non-pathogenic coliforms may be partly especial y Shigel a. Dif erentiation of Shigel a and Salmonel a inhibited or show large, flat, yel ow colonies. Shigel a from non-pathogenic bacteria is accomplished by three produces red colonies, as do hydrogen sulfide-negative reactions: xylose fermentation, lysine decarboxylation, and salmonel ae. Hydrogen sulfide producing salmonel a hydrogen sulfide production. grow as red colonies with black centers. Rose Bengal Chloramphenicol Agar is recommended for the
selective isolation and enumeration of yeasts and moulds from environmental materials and foodstuf s. Mycological peptone or papaic digest of soyabean meal provides essential growth Rose Bengal Chloramphenicol Agar after 5 days at 20-
nutrients. Rose Bengal dye suppresses the development of 25 o C produces good to excel ent growth of bacteria and reduces the spreading of moulds, control the size Saccharomyces cerevisiae and Candida albicans with and height of mould colonies such as Rhisopus species. total inhibition of E. coli. Chloramphenicol has inhibitory action on gram-negative Salmonel a ID Agar. Developed for the improved isolation of
Salmonel a spp. from food, Salmonel a ID Agar utilizes a dual Based on DCA Hynes, Salmonel ae turn green as they chromogenic technique to give a simple but ef ective color can only metabolize the X-alfa-gal chromogen, whereas change to dif erentiate Salmonel a from al other al other Enterobacteriaceaeae turn black due to their ability to metabolize both chromogens. DISPOSAL
The used NOVASTREAK MCMD is disposed by standard methods of biohazard disposal.
EXPIRATION DATE
1. The expiration date applies to the product in its intact container when stored as directed.
2. Do not use NOVASTREAK MCMD exhibiting any of the fol owing characteristics: discoloration, dehydration,
wrinkling or shrinkage of an agar surface; microbial growth prior to inoculation; or an atypical cultural response in Quality Control procedures.

INCUBATION CONDITIONS
REAGENTS
TEMPERATURE
PRELIMINARY
RESULTS (Hrs)
Total Bacterial Count Total Coliforms Baird Parker Agar
Salmonel a, Shigel a Rose Bengal
Salmonel a ID

INTERPRETATION OF RESULTS
1. BACTERIAL COUNT
If more than 200 colonies grow on the NOVASTREAK MCMD, the growth may become semi-confluent and the presence of more than 100,000 bacteria per ml is indicated. If fewer than 20 colonies are counted, less than 10,000 bacteria per ml is indicated.

COLONY DENSITY CHART
NOVASTREAK Microbial Contamination Monitoring Device
Colony Density Chart No. 1: SAMPLING BY STREAKING
Number of colonies
11-24 25-50
Colony Density Chart No. 2: SAMPLING BY DIPPING
Number of colonies
Colony Density Chart No. 3: TOTAL GROWTH BY SURFACE CONTACT SAMPLING
Number of colonies up to 3
Colony Density Chart No. 4:YEAST GROWTH BY
Colony Density Chart No. 5: MOLD GROWTH BY
SURFACE CONTACT SAMPLING.
SURFACE CONTACT SAMPLING.
Moderate
Moderate
(up to 1 colony) (up to 10 colonies) (confluent growth)
(up to 1 colony)
(up to 10 colonies)
(confluent growth)
2. COLONIES MORPHOLOGY
Preliminary identification of the microorganisms made on the base of type and color of the colonies. S.typhimurium Colorless No growth Colorless No growth
3. MICROBIOLOGICAL CRITERIA
Mandatory microbiological criteria have been established by such bodies as the U.S. Public Health Service, the USDA, the ICMSF, etc. [1-4]. Test parameters for raw milk cal for an aerobic plate count using a plate count agar (PCA)/standard methods agar (STM). For Grade A Pasteurized (uncultured and cultured milk) aerobic bacteria and coliforms must be tested. Frozen desserts must be similarly tested for total aerobes and coliforms. Breaded shrimps must be tested for total aerobes, E. coli, S. aureus. In other countries caseins and caseinates must be tested for total aerobic microorganisms, thermophilic organisms and coliforms. Codex criteria for natural mineral waters include an aerobic count of mesophiles, coliforms, E. coli, enterococci, sulphite reducing anaerobes, P. aeruginosa, and other pathogenic organisms. In addition, Canadian criteria cal for a mold count in tomato juice, a total plate count and E. coli count for fish protein, as wel as a total plate count, coliform and Salmonel a counts for gelatin. Advisory microbiological criteria have also been established by ICMSF and the Codex Commission [5- 6] for, among others, the fol owing food products: Roast beef as wel as pate should be tested for Salmonel a. On raw chicken an aerobic plate count should be performed. Cooked poultry (frozen) should be tested for S. aureus and Salmonel a. Fresh and frozen fish (to be cooked) should be tested for total aerobes, E. coli, Vibrio parahaemolyticus, S. aureus and Salmonel a. Dried milk should be tested for total aerobes, coliforms and Salmonel a (for normal and high risk populations). Fresh cheese and soft cheese should be tested for S. aureus and coliforms. Egg products (pasteurized liquid, frozen and dried) should be tested for total aerobes, coliforms, and Salmonel a (for normal and high risk populations). Chocolate/confectionery, as wel as coconut should be tested for Salmonel a. References:
1. Public Health Service-Food and Drug Administration. 1978. Grade A Pasteurized Milk Ordinance. 1978 Recommendations. PHS/FDA Publication No. 229. U.S. Government Printing Of ice, Washington, D.C. 2. Department of Agriculture. 1975. General specifications for approved dairy plants and standards for grades of dairy products. Fed. Regist. 40(198):47910-47940. 3. Food and Drug Administration. 1989. Raw breaded shrimp-microbiological criteria for evaluating compliance with current good manufacturing practice regulations, chapter 8. In: Compliance Policy Guides 7108.25. Food and Drug Administration, Washington, 4. National Research Council. 1985. An Evaluation of the Role of Microbiological Criteria for Foods and Food Ingredients. National Academic Press, Washington, D.C. 5. International Commission on Microbiological Specifications for Food. 1986. Microorganisms in Foods: Sampling for microbiological Analysis: Principles and Applications. 2nd ed. University of Toronto Press. Toronto. 6. Codex Alimentarius Commission, 20th session. 1993. Report of the 20th Session of the Codex Commission on Food Hygiene. Alinorm 93/13A. Food and Agriculture Administration, Rome.

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