NOVASTREAK Microbial Contamination Monitoring Device

Transcription

1 28 PIERRE KÖENIG ST., TALPIOT INDUSTRIAL AREA POB JERUSALEM ISRAEL TEL FAX NOVASTREAK Microbial 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 skilled personnel then plate it. Since food is an excellent 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 Monitoring Device (MCMD) is your superior device, designed to be one step ahead of all culturing devices and to further reduce hands-on involvement. The unique dip-tip technique differentiates NOVASTREAK MCMD from all 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 10 7 organisms per milliliter. NOVASTREAK MCMD is intended for use in the food industry. SUMMARY AND EXPLANATION NOVASTREAK MCMD comprises a plastic paddle with two types of agar attached back-to-back, housed in a closed transparent plastic tube. A ring with elongated prongs is attached 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 reinsertion 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 10 7 organisms per milliliter. NOVASTREAK MCMD can be used to monitor microbial wherever the potential may exceed 10 3 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 different agar modifications attached back-to-back on a plastic sampling paddle, which is permanently fastened to the cap for comfort of handling during use. NOVASTREAK MCMD are selfcontained 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 specific media and color indicators. ADVANTAGES OF METHOD The total extra-cost for TCM include: (1) expensive skilled 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. 1

2 NOVASTREAK MCMD is a leading product in comparison to the TCM with the following advantages: (1) easy protocol simple, rapid and handy, (2) no skilled personnel are required due to the simple to use straightforward device, (3) sampling at the site of collection 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 READING Liquid samples Viscous and friable samples Solid samples Surfaces Milk (raw and pasteurized) Industrial water (waste, recycled, cooling or process water), dairy products (starter cultures, sour cream, yogurt and other fermented products) Syrups, pasts and dehydrated products (vegetables, fruit, egg powder, milk powder, powdered soups, instant desserts, cocoa, etc.) Raw material, frozen and chilled products ( meat, fish and sea food products) Utensils, work surfaces Dip sampling procedure General streaking procedure Dilute 1:1 or 1:10 in sterile water with following General streaking procedure Homogenize and suspended 1:1 or 1:10 in sterile water with following General streaking procedure Touch surface with paddle for several seconds Chart. 2 Chart. 1 Chart. 1 and multiply the result by 2 or 10 Chart. 1 and multiply the result by 2 or 10 Chart. 3 GENERAL PROCEDURE A. STREAKING SAMPLING 1. Unscrew the NOVASTREAK MCMD cap. Pull the paddle out. Do not touch any part but the cap. 2. Hold the paddle vertically and dip the white prongs into the sample up to about half of their length (see below). 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 (35 o C±2 o C) for hours in a vertical position. 7. Interpret the results by simple visual comparison of bacterial on the agar surface with Colony Density Chart. 1 provided. actual colony counting is necessary. B. DIP SAMPLING (DIPSLIDE TECHNIQUE) 1. Unscrew the NOVASTREAK MCMD cap. Pull 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 pull 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 fully 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 (35 o C±2 o C) for hours. Before incubation, loosen cap one-half turn. 7. Interpret the results by simple visual comparison of bacterial on the agar surface with Colony Density Chart. 2 provided. actual colony counting is necessary. C. SURFACE CONTACT SAMPLING 1. Unscrew the NOVASTREAK MCMD cap. Pull 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 pull 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. 2

3 5. Transport NOVASTREAK MCMD vial to laboratory for incubation and examination. 6. Place inoculated NOVASTREAK MCMD vial upright in incubator (35 o C±2 o C) for hours. Before incubation, loosen cap one-half turn. 7. Interpret the results by simple visual comparison of bacterial on the agar surface with Colony Density Chart. 3 provided. actual colony counting is necessary. MATERIALS PROVIDED CAT. BD-504 BD-506 BD-507 BD-508 BD-511 BD-521 Side 1: Plate Count Agar (PCA) Side 2: Oxytetracycline-Glucose-Yeast Extract Agar (OGYE) Side 1: Violet Red Bile Agar (VRBA) Side 2: Ox tetracycline-glucose-yeast Extract Agar (OGYE) Side 1: Violet Red Bile Agar (VRBA) Side 2: Plate Count Agar (PCA) Side 1: Baird Parker Agar Side 2: Violet Red Bile Agar (VRBA) Side 1: Oxytetracycline-Glucose-Yeast- Extract Agar (OGYE) Side 2: Rose Bengal Agar w.chloramphenicol Side 1: XLD Agar Side 2: Salmonella ID Agar COLOR OF Agar Color: Purple Agar Color: Purple, Opaque Agar Color: Purple Agar Color: Pink Agar Color: Orange EXPECTED TYPES OF MICROORGANISMS Total Bacterial Count Total Coliforms Count Total Coliforms Count Total Bacterial Count Staphylococci Total Coliforms Count Salmonella, Shigella Salmonella APPLICATION FIELDS MATERIAL TO BE TESTED Total Coliforms Count TYPE OF MICROORGANISMS Total Bacterial Count Yeast & Molds Water (waste, recycled, cooling or process water) Raw Milk Dairy Products (pasteurized milk, starter cultures, sour cream, yogurts and other fermented dairy products) Meat, fish, sea food (raw material, frozen and chilled 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 material. STORAGE 1. Store NOVASTREAK MCMD at 2-8 o C 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 3

4 KIT CONTENTS 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 enumeration of bacterial and fungal microorganisms of milk and other dairy products, and may be used to determine the 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 foodstuffs. The medium uses oxytetracycline as the selective agent at a neutral ph, that gives increased counts of yeasts and molds compared with media, which rely on a low ph to suppress bacterial. Violet Red Bile Agar (VRBA) is a selective medium for the detection of coliform organisms in water, milk, and other materials of sanitary importance. The medium is selective due to the presence of inhibitors, bile salts and crystal violet. Differentiation of enteric microorganisms is achieved by the combination of lactose and the neutral red indicator. Colorless or pink-to-red colonies are produced depending on the ability to ferment lactose. Baird Parker Agar is used for the selective isolation and enumeration of coagulase-positive staphylococci from food, skin, soil, air and other materials. It may also be used to identify staphylococci on the basis of their ability to clear egg yolk (lecithinase production). Sodium pyruvate is incorporated to impart S. aureus their black colony appearance. Glycine and lithium chloride have inhibitory action on organisms other than S. aureus. Xylose Lysine Decarboxylase (XLD) Agar is recommended for the isolation and differentiation of enteric pathogens, especially Shigella. Differentiation of Shigella and Salmonella from non-pathogenic bacteria is accomplished by three reactions: xylose fermentation, lysine decarboxylation, and hydrogen sulfide production. Rose Bengal Chloramphenicol Agar is recommended for the selective isolation and enumeration of yeasts and moulds from environmental materials and foodstuffs. Mycological peptone or papaic digest of soyabean meal provides essential nutrients. Rose Bengal dye suppresses the development of bacteria and reduces the spreading of moulds, control the size and height of mould colonies such as Rhisopus species. Chloramphenicol has inhibitory action on gram-negative bacteria. Salmonella ID Agar. Developed for the improved isolation of Salmonella spp. from food, Salmonella ID Agar utilizes a dual chromogenic technique to give a simple but effective color change to differentiate Salmonella from all other Enterobacteriaceaeae. EXPECTED RESULTS A variety of bacterial and fungal species appearing as contaminants in dairy and other food products will show good, including lactobacilli and staphylococci. OGYE agar, after 5 days at o C produces good to excellent of Aspergillus niger, Saccharomyces cerevisiae, with total inhibition of E. coli. Lactose fermenters are rose-red in color and generally surrounded by a halo of precipitated bile. E. coli colonies are entire edged, 1mm or more in diameter. E. aerogenes are larger, often mucoid and pinkish. Lactose non-fermenters produce colorless colonies. Enterococci occasionally grow to produce rose colonies pinpoint in size Typical colonies of S. aureus are black, shiny, convex and surrounded by clear halos of approximately 2 to 5 mm. Coagulase-negative staphylococci do not grow well; if at all, the typical clear halos are absent. Other organisms are inhibited or may grow sparsely, producing white to brown colonies with no clearing of the egg yolk. E. coli and non-pathogenic coliforms may be partly inhibited or show large, flat, yellow colonies. Shigella produces red colonies, as do hydrogen sulfide-negative salmonellae. Hydrogen sulfide producing salmonella grow as red colonies with black centers. Rose Bengal Chloramphenicol Agar after 5 days at o C produces good to excellent of Saccharomyces cerevisiae and Candida albicans with total inhibition of E. coli. Based on DCA Hynes, Salmonellae turn green as they can only metabolize the X-alfa-gal chromogen, whereas all other Enterobacteriaceaeae turn black due to their ability to metabolize both chromogens. 4

5 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 following characteristics: discoloration, dehydration, wrinkling or shrinkage of an agar surface; microbial prior to inoculation; or an atypical cultural response in Quality Control procedures. INCUBATION CONDITIONS OBJECT TEMPERATURE ( o C) PRELIMINARY RESULTS (Hrs) FINAL RESULTS (Hrs) PCA Total Bacterial Count OGYE VRBA Total Coliforms Count Baird Parker Agar Staphylococci XLD Agar Salmonella, Shigella Rose Bengal Chloramphenicol Agar Salmonella ID Agar Salmonella INTERPRETATION OF RESULTS 1. BACTERIAL COUNT If more than 200 colonies grow on the NOVASTREAK MCMD, the 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. 5

6 COLONY DENSITY CHART NOVASTREAK Microbial Monitoring Device Colony Density Chart. 1: SAMPLING BY STREAKING Number of colonies up to over 50 CFU/ml Colony Density Chart. 2: SAMPLING BY DIPPING Number of colonies up to over 50 CFU/ml Colony Density Chart. 3: TOTAL GROWTH BY SURFACE CONTACT SAMPLING Number of colonies up to over 50 CFU/ml Colony Density Chart. 4:YEAST GROWTH BY SURFACE CONTACT SAMPLING. Colony Density Chart. 5: MOLD GROWTH BY SURFACE CONTACT SAMPLING. Light (up to 1 colony) Moderate (up to 10 colonies) Heavy (confluent ) Light (up to 1 colony) Moderate (up to 10 colonies) Heavy (confluent ) 6

7 2. COLONIES MORPHOLOGY Preliminary identification of the microorganisms made on the base of type and color of the colonies. ORGANISMS VRBA OGYE PCA Baird - Parker XLD Rose Bengal Chloramphenicol Agar Salmonella ID E.coli Red White Yellow or none Black Coliforms Red or Yellow White pink or none Black Red S.typhimurium Colorless White w/black Green center S.flexneri Colorless White Red Colorless S.aureus Black White w/halo S.epidermidis Black White small C.albicans White White Pink 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 call 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 call for a mold count in tomato juice, a total plate count and E. coli count for fish protein, as well as a total plate count, coliform and Salmonella counts for gelatin. Advisory microbiological criteria have also been established by ICMSF and the Codex Commission [5-6] for, among others, the following food products: Roast beef as well as pate should be tested for Salmonella. On raw chicken an aerobic plate count should be performed. Cooked poultry (frozen) should be tested for S. aureus and Salmonella. Fresh and frozen fish (to be cooked) should be tested for total aerobes, E. coli, Vibrio parahaemolyticus, S. aureus and Salmonella. Dried milk should be tested for total aerobes, coliforms and Salmonella (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 Salmonella (for normal and high risk populations). Chocolate/confectionery, as well as coconut should be tested for Salmonella. References: 1. Public Health Service-Food and Drug Administration Grade A Pasteurized Milk Ordinance Recommendations. PHS/FDA Publication U.S. Government Printing Office, Washington, D.C. 2. Department of Agriculture General specifications for approved dairy plants and standards for grades of dairy products. Fed. Regist. 40(198): Food and Drug Administration Raw breaded shrimp-microbiological criteria for evaluating compliance with current good manufacturing practice regulations, chapter 8. In: Compliance Policy Guides Food and Drug Administration, Washington, D.C. 4. National Research Council 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 Microorganisms in Foods: Sampling for microbiological Analysis: Principles and Applications. 2nd ed. University of Toronto Press. Toronto. 6. Codex Alimentarius Commission, 20th session Report of the 20th Session of the Codex Commission on Food Hygiene. Alinorm 93/13A. Food and Agriculture Administration, Rome. 7