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In This Issue 1 TechniTopics

· Foodborne Diseases An Update, Part II · The Gram Stain Revisited

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BD Lab·O

Microbiology News & Ideas

Spring 2005 · Volume 16 · No. 1

BACTECTM System News · BACTECTM 9000 Culture Club Product Highlights · BD EpiCenterTM Version 5 Software · BD CultureSwabTM MaxV - Maximizing Recovery of Microorganisms · BBLTM CHROMagarTM MRSA - The New Standard in Rapid MRSA Screening · BBLTM Media Enhancements · BD DifcoTM Antisera - Salmonella and Shigella Grouping Sets

Foodborne Diseases

AN UPDATE, PART II

Kenneth D. Wilde, Ph.D.

12 FYI

· Nasopharyngeal Specimen Collection Wall Chart · Get Clued to the Flu · Launching - Our Newly Reconfigured Web Site · 2005 Customer Training Calendar · The Industrial and Clinical Media Advisory Team

15 Micro Happenings

· FIND and BD Combine Efforts for Rapid TB Diagnosis · 105th ASM General Meeting

Part I of this two-part article, published in the previous issue of LabOTM (Vol. 15, No. 3), introduced the CDC's Foodborne Diseases Active Surveillance Network (FoodNet) and its intention to reduce the incidence of the four most frequently reported foodborne pathogens to the projected levels stated by the 2010 national health objectives. The four pathogens and their projected 2010 levels include Campylobacter jejuni (12.3 per 100,000), various serovars of Salmonella enteritidis (6.8 per 100,000), Escherichia coli O157:H7 (1.0 per 100,000) and Listeria monocytogenes (0.25 per 100,000).1 In Part I, we focused on Campylobacter jejuni, the leading cause of bacterial diarrheal illness in the U.S. In Part II, we will focus on

Listeria monocytogenes on Oxford Medium

Listeria monocytogenes, arguably the most serious and devastating of the foodborne pathogens.

Listeria monocytogenes

Listeria monocytogenes' ubiquity and unique ability to persevere in unfavorable conditions and refrigerated temperatures makes it a problematic organism for food microbiologists.2 The bacterium was initially named Bacterium monocytogenes because it caused a mononuclear leucocytosis in rabbits. The change in its genus nomenclature appeared a few years later. It was not until 1985 that L. monocytogenes became recognized as a formidable human pathogen when it was identified as the etiologic agent of a severe foodborne outbreak involving Mexican-style soft cheese (142 cases with 48 deaths).3

Continued on page 2

Computer-assisted laboratory methods are used to record growth of L. monocytogenes bacteria on ready-to-eat meat products.

Electron micrograph of Listeria, courtesy of CDC/Dr. Balasubr Swaminathan; Peggy Hayes.

Photo by Paul Pierlott, courtesy of the Agricultural Research Service.

TechniTopics

Foodborne Diseases

Continued from page 1

Listeriosis is the name of the general group of disorders caused by L. monocytogenes. Listeria organisms are widely disseminated in the rural environment and consequently contaminate both the raw materials used in the preparation of industrially processed foods and the surfaces in food production plants. L. monocytogenes is a psychrotroph and is, therefore, a concern in the extended shelf-life of refrigerated foods. The ability of the organism to grow over a wide temperature range and in the presence or absence of oxygen enables it to multiply in many environments.4 This makes Listeria a serious threat to food safety and ranks it among the microorganisms that most concern the food industry. L. monocytogenes is a pleomorphic, nonsporeforming, nonencapsulated, gram-positive facultative anaerobic rod that ranges in size from 0.2 to 0.4 µ in diameter to 0.5 to 2.0 µ in length.2,5 Its motility occurs between 20 to 25°C (but not at 37°C) and exhibits a characteristic umbrella-like subsurface growth in semi-solid media (i.e., Motility Medium). Cellular arrangement may be either singly, in pairs or short chains with optimal proliferation occurring from pH 7 to slightly alkaline and within a temperature range of 25 to 36°C. Moreover,

this pathogen possesses the unique capabilities of growing up to a pH of 9.6, as well as in the presence of 10% sodium chloride. Other diagnostic characteristics denote that L. monocytogenes is catalase positive and oxidase

L. monocytogenes has been associated with foods such as raw milk, insufficiently pasteurized fluid milk, cheeses (particularly soft-ripened varieties), ice cream, raw vegetables, raw and cooked poultry, all types of raw meats and raw and smoked fish.

enrichment broth, which contains the sample, is incubated for 24 hours at 30°C. An aliquot is then transferred to a secondary enrichment broth (Fraser Broth), which, after 24 hours, is inoculated onto LPM Agar. LPM Agar is an improved selective medium containing the ingredients lithium chloride, phenylethanol and moxalactam, a broad-spectrum antibiotic which suppresses the growth of both grampositive and gram-negative bacteria.3 Rapid detection of L. monocytogenes as well as Listeria species in foods is now frequently achieved using PCR methodology (e.g., BAXTM System, Qualicon, Inc.). Since this method detects only the presence of L. monocytogenes DNA, isolation of the organism is needed for confirmation. Incidence data for 1987, prospectively collected by CDC, suggested that there were at least 1,600 cases of listeriosis with 415 deaths per year in the U.S. The vast majority of cases are considered sporadic, making epidemiologic links to particular food items very difficult. The main target population for Listeria infections include: pregnant women and the fetus; immunocompromised persons such as cancer patients (leukemia patients in particular); the elderly; and normal, healthy people who have been exposed to heavily contaminated food or beverage items.6 L. monocytogenes has been associated with foods such as raw milk, insuffiContinued on page 16

negative. After 24 hours on primary blood agar, colonies of this organism appear small (about 1 mm in diameter) smooth, round and translucent. The colonies are surrounded by a narrow band of beta-hemolysis which becomes more evident when the colony is removed. This hemolytic activity can be verified by the CAMP test in conjunction with Staphylococcus aureus.5 Recovery of the organism from foods is perhaps more challenging and should follow the USDA protocol recommended for isolating the organism from meat and poultry. A primary

Kenneth D. Wilde, Ph.D.

Dr. Wilde recently retired from the Maryland Department of Health and Mental Hygiene, where he served as the Chief, Division of Public Health Environmental Microbiology. Following receipt of a B.A. in Natural Science from Kutztown University, he earned M.S. and Ph.D. degrees in Microbiology from the University of Maryland.

His duties at the department of health involved the areas of food/shellfish safety, dairy and water bacteriology, including serving as State Water Quality Laboratory Certification Officer, and overseeing the operations of the food/water bioterrorism laboratory. He was entrusted to analyze powder samples for anthrax spores during the Fall and Winter 20012002 and participated in the presentation of numerous state laboratory-sponsored bioterrorism workshops. Dr. Wilde is a certified Specialist Microbiologist in Public Health and Medical Laboratory Microbiology with the National Registry of Microbiologists of the American Academy of Microbiology.

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BD Lab·O

volume 16 · number 1

TechniTopics

The Gram Stain Revisited

The Gram stain is one of the oldest and most important diagnostic procedures performed in the clinical microbiology laboratory. Performing this stain correctly on clinical specimens helps microbiologists and physicians with rapid, valuable information as to whether infection is present, and if so, to categorize the infection. The Gram stain is clearly a valuable diagnostic assay, but caution needs to be exercised both in its application and in the interpretation of results to prevent the dissemination of misleading information. Quite simply, a Gram stain may reveal the presence or absence of microorganisms, and either result may or may not be significant or indicative of infection. False-negative Gram stains can occur in infectious diseases that are associated false-positive Gram stain is the presence of contaminating microorganisms in the specimen, such as a sputum specimen. Less commonly recognized causes of a false-positive Gram stain are the presence of artifacts, which may be in one or more of the different reagents necessary to conduct the stain. Artifacts may also be present in the specimen, on the slide, in the immersion oil, in the system used to transport the specimen (agar-based transport swabs), or in the microbiological culture medium used to grow the microorganisms.

of methods are used to reduce and or eliminate the bioburden of microorganisms in our microbiological culture media.3 These methods include terminal moist-heat sterilization (autoclaving), irradiation and filtration (performed in conjunction with aseptic filling). The primary objective of autoclaving is to provide the appropriate amount of heat that will result in a low probability of non-sterility yet not adversely affect the appearance or performance of the product. This can be a challenge with media formulations that have high levels of simple sugars (e.g., glucose) and amino acids from proteins (e.g., lysine) that react under heat, resulting in Maillard browning4 and an undesirable appearance. Although autoclaving reduces the viable bioburden, the process does not remove the microorganisms destroyed in the process or their remains, particularly nonviable whole cells, cell wall fragments, toxins and other proteins. These remains become the recognizable artifacts visible following a simple staining procedure such as the Gram stain. These nonviable structures in the medium may also contribute to a falsepositive Gram stain result.

The Gram stain is clearly a valuable diagnostic assay, but caution needs to be exercised both in its application and in the interpretation of results to prevent the dissemination of misleading information.

with low ("paucibacillary") microbial load. In these situations, a "negative" Gram stain result may not be equivalent to "no infection." One frequent example is that of prosthetic joint infections. These infections are typically associated with very low microbial loads. Less than 10% of specimens from patients who are actually infected have a positive Gram stain.1 In other words, 90% or more of patients with a definitive prosthetic joint infection will have a negative Gram stain result. So, using a negative Gram stain result to "rule-out" infection is not possible. In addition, the Gram stain is not 100% specific for microorganisms or infection and, therefore, cannot be used to conclusively "rule-in" infection as well. A commonly recognized cause of a If used inappropriately, liquid culture media (e.g. Tryptic Soy Broth, Brain Heart Infusion Broth, Fluid Thioglycollate Medium) may produce false-positive Gram stain results due to the presence of nonviable microorganisms that arise from the initial bioburden population of microorganisms in the product. A number of sources may contribute to the level of artifacts or nonviable bioburden level including: biological components of the dehydrated culture medium; water; packaging (e.g., tubes, bottles, caps); processing environment; processing equipment; and people who come in contact with the product. Sterilization is a process designed to eliminate viable microorganisms from a material or medium.2 At BD, a variety

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BD Lab·O

volume 16 · number 1

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BD BACTEC TM System News

The BACTECTM 9000 Culture Club Has New Members and a New List!

The BACTECTM 9000 Culture Club was created to inform BACTEC 9000 System users of unusual organisms recovered from BACTEC 9000 series instruments and BACTEC media. As newly isolated organisms are reported to us, the reports are published in LabOTM. At this time, we are pleased to add 13 new "members of the club." In addition, we have prepared an updated list of all the organisms recovered on the BACTEC 9000 series instruments. Since the last list was published in June 2002, 36 organisms have been added for a grand total of 272 organisms! Why don't you "join" the club? If you have an unusual organism isolated from any of the BACTEC 9000 series instruments (9240, 9120, 9050 and 9000MB), see if it is listed on the BACTEC 9000 Culture Club form on page 5. If it's not listed, complete the form on page 6 and send it in to receive your complimentary BD Portfolio! There is no limit to the number of forms that can be submitted. For more information on the BACTECTM 9000 Culture Club or to obtain additional forms, call BD Technical Services at 800.638.8663. * BD Diagnostics does not claim recovery of the isolates listed in the table with the associated media. See package inserts for the expected organism recovery.

Laboratory Site Beloit Memorial Hospital Beloit, WI

Underlying Disease/ Diagnosis Bartter's Syndrome/ Gitelman's Syndrome Endocarditis

BACTEC Instrument 9240

BACTEC Media* Lytic 10/ Anaerobic/F

Time to Detection 93 hours

Organism Detected Clostridium difficile

Community Medical Center Scranton, PA

9240

Standard Aerobic/F Standard Aerobic/F Peds Plus Aerobic Plus Aerobic/F Plus Aerobic/F

5 days

Rothia dentocariosa Mycobacterium neoaurum Brucella suis

FUO

9240

4 days

North Florida Regional Medical Center Gainesville, FL

Infected hematoma Dehydration/ bronchitis Pneumonia

9240

70 hours

9240

45 hours

Methylobacterium sp. Actinomyces viscosus

Peninsula Regional Medical Center Salisbury, MD Riverview Hospital Noblesville, IN

9240

93 hours

Abdominal surgery and ileostomy Unknown

9120

Plus Aerobic/F

24 hours

Rhizobium (Agrobacterium) radiobacter Brevibacterium casei Dermabacter hominis Corynebacterium striatum Cellulomonas sp. Weissella confusa Arthrobacter sp.

Valley Baptist Medical Center Harlingen, TX

9240

Peds Plus Aerobic Standard Anaerobic/F Peds Plus Aerobic Standard Aerobic/F Peds Plus Aerobic Peds Plus Aerobic

48 hours

Congestive Heart Failure Febrile illness Medulloblastoma Sepsis Bronchiolitis

9240 9240 9240 9240 9240

4 days 48 hours 24 hours 24 hours 48 hours

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BD Lab·O

volume 16 · number 1

BD BACTEC TM System News

BACTECTM 9000 Culture Club

Aerobic Gram-Negative Organisms

Acinetobacter baumannii Acinetobacter lwoffi Acinetobacter sp. Aeromonas hydrophila Aeromonas sp. Alcaligenes faecalis Alcaligenes xylosoxidans subsp. xylosoxidans Bordetella pertussis Brevundimonas diminuta Brevundimonas vesicularis Brucella suis Brucella sp. Burkholderia cepacia Burkholderia pickettii Burkholderia pseudomallei Campylobacter fetus Campylobacter jejuni subsp. jejuni Campylobacter sp. Capnocytophaga canimorsus Capnocytophaga cynodegmi Cardiobacterium hominis Chromobacterium violaceum Chryseobacterium indologenes Chryseobacterium meningosepticum Chryseomonas luteola Citrobacter amalonaticus Citrobacter diversus Citrobacter freundii Citrobacter sp. Comamonas acidovorans Eikenella corrodens Enterobacter aerogenes Enterobacter agglomerans Enterobacter cancerogenus Enterobacter cloacae Enterobacter sakazakii Escherichia coli Escherichia vulneris Flavobacterium sp. Haemophilus aphrophilus Haemophilus influenzae Haemophilus parainfluenzae Haemophilus paraphrophilus Haemophilus sp. Helicobacter rappini Kingella kingae Kingella sp. Klebsiella oxytoca Klebsiella ozaenae Klebsiella pneumoniae Kluyvera ascorbata Kluyvera sp. Methylobacterium mesophilicum Methylobacterium sp. Moraxella catarrhalis Moraxella nonliquefaciens Moraxella sp. Morganella morganii Neisseria gonorrhoeae Neisseria meningitidis Neisseria mucosa Neisseria sicca Neisseria subflava Neisseria sp. Ochrobactrum anthropi Pasteurella multocida Plesiomonas shigelloides Proteus mirabilis Proteus penneri Proteus vulgaris Providencia alcalifaciens Providencia rettgeri Providencia stuartii Pseudomonas aeruginosa Pseudomonas fluorescens Pseudomonas fluorescens group Psychrobacter immobilis Rhizobium (Agrobacterium) radiobacter Riemerella anatipestifer Roseomonas sp. Salmonella arizonae Salmonella paratyphi A Salmonella typhi Salmonella serotype Montevideo Salmonella sp., Grp B Salmonella sp., Grp D Salmonella sp., Grp G Salmonella sp. Serratia liquefaciens Serratia marcescens Serratia plymuthica Shewanella putrefaciens Shigella sonnei Shigella sp. Sphingomonas paucimobilis Stenotrophomonas maltophilia Streptobacillus moniliformis Vibrio cholerae, non-01 Vibrio fluvialis Vibrio mimicus Vibrio vulnificus Yersinia enterocolitica Bacillus licheniformis Bacillus subtilis Bacillus sp. Brevibacterium casei Brevibacterium paucivorans Cellulomonas sp. Corynebacterium Grp B-1 Corynebacterium Grp D-2 Corynebacterium aquaticum Corynebacterium jeikeium Corynebacterium striatum Corynebacterium xerosis Corynebacterium sp. Dermabacter hominis Enterococcus avium Enterococcus casseliflavus Enterococcus durans Enterococcus faecalis Enterococcus faecium Enterococcus gallinarum Enterococcus sp. Erysipelothrix rhusiopathiae Facklamia hominis Gemella morbillorum Lactococcus sp. Leuconostoc sp. Listeria monocytogenes Micrococcus sp. Oerskovia sp. Pediococcus sp. Rhodococcus equi Rhodococcus sp. Roseomonas sp. Rothia dentocariosa Rothia mucilaginosa Staphylococcus aureus Staphylococcus auricularis Staphylococcus capitis Staphylococcus cohnii Staphylococcus epidermidis Staphylococcus haemolyticus Staphylococcus hominis Staphylococcus lugdunensis Staphylococcus saccharolyticus Staphylococcus saprophyticus Staphylococcus schleiferi Staphylococcus sciuri Staphylococcus simulans Staphylococcus warneri Staphylococcus xylosus Stomatococcus mucilaginosus Stomatococcus sp. Streptococcus acidominimus Streptococcus anginosus Streptococcus bovis Streptococcus constellatus Streptococcus intermedius Streptococcus milleri Streptococcus mitis Streptococcus mutans Streptococcus pneumoniae Streptococcus salivarius Streptococcus sanguis Streptococcus uberis Streptococcus vestibularis Streptococcus Grp A Streptococcus Grp B Streptococcus Grp C Streptococcus Grp D Streptococcus Grp F Streptococcus Grp G Streptococcus sp., gamma hemolytic Streptococcus sp., nutritionally variant Streptococcus sp., viridans Weissella confusa Peptostreptococcus prevotii Peptostreptococcus sp. Porphyromonas endodontalis Porphyromonas gingivalis Prevotella bivia Prevotella buccae Prevotella oralis Prevotella oris Propionibacterium acnes Propionibacterium sp. Streptobacillus moniliformis Veillonella parvula Veillonella sp.

Yeast/Fungi Organisms

Alternaria sp. Aspergillus niger Candida albicans Candida guilliermondii Candida kefyr Candida krusei Candida lusitaniae Candida parapsilosis Candida stellatoidea Candida tropicalis Coccidioides immitis Cryptococcus neoformans Fusarium sp. Histoplasma capsulatum Malassezia furfur Prototheca sp. Rhodotorula rubra Rhodotorula sp. Torulopsis glabrata Trichophyton rubrum

Anaerobic Organisms

Anaerobiospirillum succiniciproducens Bacteroides caccae Bacteroides fragilis Bacteroides fragilis group Bacteroides melaninogenicus Bacteroides ovatus Bacteroides splanchnicus Bacteroides thetaiotaomicron Bacteroides ureolyticus Bacteroides vulgatus Bacteroides sp. Bifidobacterium sp. Clostridium butyricum Clostridium clostridioforme Clostridium difficile Clostridium innocuum Clostridium limosum Clostridium paraperfringens Clostridium perfringens Clostridium ramosum Clostridium septicum Clostridium sporogenes Clostridium tertium Clostridium sp. Eubacterium lentum Eubacterium limosum Eubacterium sp. Fusobacterium mortiferum Fusobacterium necrophorum Fusobacterium nucleatum Fusobacterium varium Fusobacterium sp. Lactobacillus casei Lactobacillus sp. Leptotrichia buccalis Peptococcus sp. Peptostreptococcus anaerobius Peptostreptococcus asaccharolyticus

Actinomycetes

Actinomyces odontolyticus Actinomyces viscosus Actinomyces sp. Nocardia asteroides Nocardia farcinica Streptomyces sp.

Mycobacteria

Mycobacterium avium-intracellulare Mycobacterium chelonae Mycobacterium flavescens Mycobacterium fortuitum Mycobacterium malmoense Mycobacterium mucogenicum Mycobacterium neoaurum Mycobacterium simiae Mycobacterium sp. Mycobacterium terrae Mycobacterium tuberculosis Mycobacterium xenopi

Aerobic Gram-Positive Organisms

Aerococcus sp.

Arthrobacter sp. Aureobacterium sp. Bacillus anthracis Bacillus cereus

Others

Leptotrichia sp.

BD Lab·O

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BD BACTEC TM System News

Recovery of Unusual Isolates with BD BACTECTM/F Systems

Laboratory Name : __________________________________ Institution Address: _________________________________ City/State/Zip: ______________________________________ Submitted By: ______________________________________ Telephone Number: _________________________________ e-mail*: ___________________________________________ Organism Identification: _________________________________________________________________ BACTEC Instrument: Isolated from this medium: 9240__________ 9120__________ 9050__________ 9000MB__________ PLUS Aerobic/F: __________________ Peds PlusTMAerobic:________________ Standard Aerobic/F: ______________ Myco/F-Sputa: ___________________ PLUS Anaerobic/F:___________________ LYTIC/10 Anaerobic/F: _______________ Standard Anaerobic/F: _______________ Myco F/Lytic: _______________________

Number of Specimens From The Patient: ____________________________ Number of BACTEC Cultures That Were Positive With The Isolate: Time to Detection (hours or days): Was Patient On Antimicrobial Therapy When Specimens Were Taken? ____________________________ ____________________________ ____________________________

PLEASE RETURN FORM TO: BD Diagnostics Marketing Manager, BACTEC, M.C. 642 7 Loveton Circle Sparks, Maryland 21152

Identification Method Used: _______________________________________________________________ Susceptibility Testing Method Used: _________________________________________________________ Underlying Disease or Diagnosis (if any): _____________________________________________________ _______________________________________________________________________________________ Chemistry/Hematology Lab Results: ________________________________________________________ _______________________________________________________________________________________ Antimicrobial Therapy (if any): _____________________________________________________________ _______________________________________________________________________________________

* The information you provide will not be shared with a third party vendor. By providing this information, I authorize Becton, Dickinson and Company to send me information via e-mail. 6 BD Lab·O

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Product Highlights

BD EpiCenterTM Version 5 Software:

Latest Software Benefits Microbiology, Infection Control and Pharmacy Departments

To meet the demand for automated systems that detect, monitor, audit and communicate information on patient infections, emerging resistance and nosocomial events, BD announces the availability of EpiCenterTM Version 5 Software. The EpiCenter Microbiology Data Management System centralizes the operation and data management for the BD BACTECTM, BD BACTECTM MGITTM 960 and BD PhoenixTM microbiology systems. For the microbiology lab, the EpiCenter System provides efficient-consistency in the production and communication of microbiology results. The EpiCenter System assists the laboratory in meeting the need for rapid microbiology on all shifts by improving the efficiency in the review of results. This enables managers to achieve staffing and quality assurance efficiencies. EpiCenter integrated features like "Pro-Active Alerts" and "Expertised Results" improve result consistency from every technologist. In addition, the EpiCenter System integrates continuous education features into the software enabling new or existing users to continually get more from the system. These capabilities ensure that the laboratory efficiently delivers the most accurate and timely results for their patients. Outside the lab, the EpiCenter System conveys real-time, patient-focused epidemiology. EpiCenter Version 5 Software has an optional "Multi-User" capability. Ancillary departments, such as Infection Control and Pharmacy, can provide direct patient benefits while increasing their efficiency by using this secure, direct access to the Microbiology Department's data. As an example, Infection Control Officers can effectively monitor and investigate emerging resistance and nosocomial events. Using another EpiCenter Version 5 optional module, BD EpiCARETM (EpiCenter Clinical Application Rules Editor), these departments can also implement quality assurance procedures. Infection Control Officers can construct rules that will automate policies that will assist them in meeting new Joint Commission on Accreditation of Healthcare Organizations (JCAHO) surveillance requirements. To learn more about the capabilities of the new EpiCenter Version 5 Software, contact your local BD sales representative.

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Product Highlights

Now Available ­

BD CultureSwabTM MaxV Maximizing Recovery of Microorganisms

BD CultureSwabTM MaxV, the newest addition to the BD CultureSwab specimen collection and transport product line, is specifically designed to improve recovery of microorganisms, which may improve patient diagnosis. What differentiates BD CultureSwab MaxV from other collection swabs?

Schematic courtesy of Copan Diagnostics Inc.

Reason 1: BD CultureSwab MaxV features a unique swab design consisting of hypoallergenic, non-animal proteins embedded into the rayon swab fibers to maximize organism viability. Protein provides additional nutrients for maintaining microorganisms, especially fastidious species, without creating organism overgrowth.

Reason 2: Nitrogen flushed gas-impermeable packaging seals-in the nitrogen gas, which protects the media from exposure to oxidation and moisture, assuring optimal performance.

Reason 3: Patented "Venturi-constriction" designed to maintain media moisture and minimize oxidation by forcing breaks or bubbles out of the agar, enhancing microbial survival.

Cat. No. 220231 220232 220233 220234 220235 220236

Description BD CultureSwabTM MaxV Liquid Amies, Single Swab BD CultureSwabTM MaxV Liquid Amies, Double Swab BD CultureSwabTM MaxV Liquid Stuart, Single Swab BD CultureSwab MaxV Liquid Stuart, Double Swab

TM

Qty/Pkg 50 swabs per pack 50 swabs per pack 50 swabs per pack 50 swabs per pack 50 swabs per pack 50 swabs per pack

BD CultureSwabTM MaxV(+) Amies Gel w/o Charcoal, Single Swab BD CultureSwabTM MaxV(+) Amies Gel w/o Charcoal, Double Swab

BD CultureSwab MaxV, for aerobic transport, and BD CultureSwab MaxV(+), for aerobic and anaerobic transport, can be used for collection of throat, vaginal, skin and wound specimens. For more information on BD CultureSwabTM MaxV or other BD Collection and Transport Products, mark the appropriate box(es) on the reader response card or contact your local BD sales representative.

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Product Highlights

BBLTM CHROMagarTM MRSA ­ The New Standard in Rapid MRSA Screening

We are pleased to announce that the U.S. Food and Drug Administration has cleared BBLTM CHROMagarTM MRSA medium, the new standard in rapid MRSA screening. This new prepared plated medium simplifies the process, decreases the time to result and offers high sensitivity and specificity for methicillin-resistant Staphylococcus aureus (MRSA) identification.

The prevalence of nosocomial infections caused by MRSA has been increasing for several years in many countries around the world.1 The U.S. Centers for Disease Control and Prevention estimates that between 60,000 and 80,000 Americans die each year from nosocomial infections and the cause in the majority of cases is S. aureus. BBL CHROMagar MRSA allows microbiology laboratories to identify patients colonized with MRSA more quickly and easily than the time-consuming and labor-intensive processes currently available. BBL CHROMagar MRSA allows for the direct detection and identification of most MRSA within 24 hours. The cost benefits associated with reducing nosocomial infections can be significant. "This technology will be extremely useful to those who wish to identify patients colonized with MRSA," said Dr. Bill Jarvis of Emory University School of Medicine and President, Jason and Jarvis Associates. "Since MRSA colonization leads to infection in a specific patient and is a risk factor for transmission of MRSA to other patients, rapid identification of those with MRSA colonization will reduce the risk of infection in colonized patients, by enabling their clinicians to intervene, and reduce the risk of patient-to-patient transmission by permitting isolation of those who are MRSA-colonized. Furthermore, this technology will markedly increase the rate at which community-associated or hospitalacquired MRSA patients are identified. Thus, more rapid isolation and treatment can occur. The multiple benefits of this rapid identification method will improve patient treatment, reduce the risk of transmission in healthcare settings, and reduce the burden of MRSA in U.S. hospitals." BBL CHROMagar MRSA utilizes a new chromogenic technology, which permits the detection of MRSA using chromogenic substrates and antibiotics. MRSA strains will grow in the presence of cefoxitin2 and produce mauve-colored colonies resulting from hydrolysis of the chromogenic substrate. Additional selective agents are incorporated for the suppression of gram-negative organisms, yeast and some gram-positive cocci. Bacteria other than MRSA Cat. No. may utilize other chromogenic 215084 substrates in the medium resulting in blue to blue/green colored 254102 colonies or if no chromogenic 215081 substrates are utilized, the colonies appear as white 254093 or colorless. 214982 In clinical evaluations, BBL 214983 CHROMagar MRSA displayed 3 7% greater recovery of MRSA than traditional screening algorithms and has the capability to identify MRSA earlier than most traditional algorithms. This unique technology requires less technologist time and improves the workflow in the laboratory. BBL CHROMagar MRSA allows for the direct detection and identification of most MRSA within 24 hours,

214984

or at 48 hours with a confirmatory coagulase test. For more information on BBL CHROMagar MRSA, mark the appropriate box on the reader response card.

Description BBLTM CHROMagarTM MRSA ALSO AVAILABLE BBLTM CHROMagarTM Orientation BBL CHROMagar Orientation

TM TM

Qty/Pkg 20 20 100 20 20 20 20

BBLTM CHROMagarTM Candida BBLTM CHROMagarTM Staph aureus BBL CHROMagar O157

TM TM

BBLTM CHROMagarTM Salmonella

Reacher, M.H. et al. 2000. Bacteremia and antibiotic resistance of its pathogens reported in England and Wales between 1990 and 1998:trend analysis. Br. Med. J. 320:213-6. 2 National Committee for Clinical Laboratory Standards. 2004. Performance standards for antimicrobial susceptibility testing; Fourteenth Informational Supplement, M100-S14. National Committee for Clinical Laboratory Standards, Wayne, Pa. 3 BBL CHROMagar MRSA package insert.

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Product Highlights

BBLTM Media Enhancements

As a microbiology company and a leader in media development and manufacturing, we feel it is important to routinely reassess our products and improve them. From new formulations such as the exclusive BBLTM CHROMagarTM family of products to improvements on older formulations, BD Diagnostics remains committed to providing the highest quality products for infectious disease diagnosis. Recently, we have enhanced two of our traditional media formulations, BBL Salmonella Shigella Agar and BBL Chocolate II Agar.

In both cases, we have made subtle proprietary processing and formulation adjustments to improve the product our customers require. For Salmonella Shigella Agar, the improvements made reduce the amount of naturally occurring precipitates that may become evident in this routinely used microbiological medium. Precipitation inside Salmonella Shigella Agar can sometimes resemble fungal elements embedded in the agar. These artifacts can lead some laboratories to discard Salmonella Shigella Agar needlessly, thinking the medium is contaminated. The enhancement we have made will reduce waste and discards in the laboratory. For Chocolate II Agar, we have applied our 60+ years of culture media development experience to produce a product, which we believe to be significantly different in performance than any other on the market. Growth of fastidious organisms, such as Haemophilus spp. and Neisseria spp., is rich and robust with textbook morphologies on BBL Chocolate II Agar. Reproducibility of organism morphology and consistent results are key features in this enhancement of BBL Chocolate II Agar. Contact your local BD media representative if you would like to learn more about BBL Salmonella Shigella Agar or Chocolate II Agar. We are confident that you will immediately recognize the quality and excellence that is paramount to the BBL family of media products.

Cat. No. 221181 221279 221169 221267 Description BBLTM Salmonella and Shigella Agar BBLTM Salmonella and Shigella Agar BBLTM Chocolate II Agar BBLTM Chocolate II Agar Qty/Pkg 20 100 20 100

Salmonella and Shigella Agar

Chocolate II Agar

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BD Lab·O

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Product Highlights

BD DifcoTM Antisera New ­ Salmonella & Shigella Grouping Sets

To streamline your ordering process, we are pleased to announce the immediate availability of our: · BD DifcoTM Salmonella O Grouping Antisera Set · BD DifcoTM Shigella Grouping Antisera Set Our gold standard BD Difco Antisera for the serological identification and typing of bacteria are now available in an even more convenient format than ever before. One catalog number brings you the most commonly used antisera in a single, convenient and economical package. All items may also be purchased separately. In addition, effective January 1, 2005, we reduced the list price of all BD Difco Antisera by 10%! The discounted prices are available automatically through your regular distribution channel. For more information on BD Difco Antisera for Salmonella, Shigella, Neisseria, E. coli, Vibrio, Listeria, Bordetella and Haemophilus, contact your local BD sales representative or mark the appropriate box on the reader response card.

Description Salmonella O Grouping Antisera Set contains one of each of the following: Salmonella O Antiserum Group A Factors 1, 2, 12 Salmonella O Antiserum Group B Factors 1, 4, 12, 27 Salmonella O Antiserum Group B Factors 1, 4, 5, 12 Salmonella O Antiserum Group C1 Factors 6, 7 Salmonella O Antiserum Group C2 Factors 6, 8 Salmonella O Antiserum Group D1 Factors 1, 9, 12 Salmonella O Antiserum Group E Factors 1, 3, 10, 15, 19, 34 Salmonella O Antiserum Group F Factor 11 Salmonella O Antiserum Group G Factors 13, 22, 23 (36), (37) Salmonella O Antiserum Group H Factors 1, 6, 14, 24, 25 Salmonella O Antiserum Group I Factor 16 Salmonella Vi Antiserum Salmonella O Antiserum Poly A-I & Vi Factors 1-16, 19, 22-25, 34, Vi Description Shigella Grouping Antisera Set contains one of each of the following: Shigella Antiserum Poly Group A Shigella Antiserum Poly Group A1 Shigella Antiserum Poly Group B Shigella Antiserum Poly Group C Shigella Antiserum Poly Group C1 Shigella Antiserum Poly Group C2 Shigella Antiserum Poly Group D Size Cat. No. 241107 229471 229731 229481 229491 229501 229511 228191 222601 230291 222621 222631 228271 222641 Cat. No. 241108 228341 227761 228351 228361 227771 227781 228371 3 mL 3 mL 3 mL 3 mL 3 mL 3 mL 3 mL 3 mL 3 mL 3 mL 3 mL 3mL 3 mL Size

3 mL 3 mL 3 mL 3 mL 3 mL 3 mL 3 mL

Gram Stain Revisited

Continued from page 3

media are filterable, especially those containing thickening agents like agar or gelatin (e.g., Fluid Thioglycollate Medium).

Similarly, UV and gamma irradiation will reduce the levels of viable microorganisms in a finished product, but these processes do not physically remove the nonviable microorganisms, residuals or other artifacts that are a result of the sterilization process. Sterile filtration is a useful method for sterilizing liquids and gases. Filtration physically removes viable and nonviable microorganisms from the final product rather than destroying them during the manufacturing process. After filtration, aseptic handling controls must be in place to prevent contamination from the environment, process equipment and personnel. However, not all culture

not be used for performing Gram stains directly on specimens (e.g., tissue biopsy). In addition, caution must be used when Gram staining suspicious microbial growth in liquid culture media. In conclusion, Gram stain results are affected by many variables and care must be taken in its application and interpretation.

1 Atkins, B.L., et al. 1998. Prospective evaluation of criteria for microbiological diagnosis of prosthetic-joint infection at revision arthroplasty. J. Clin. Microbiol. 36:2932. 2 Akers, J. and J. Agalloco. 1997. Sterility and sterility assurance. PDA J. Pharm. Sci. & Tech. 51:72. 3 Difco & BBL Manual. 2003. Becton, Dickinson and Company, Sparks, MD. 4 Maillard Reaction ("Browning"Reaction) L. C. Maillard, Compt. Rend. 154, 66 (1912); Ann. Chim. 9, 5, 258 (1916).

Gram stain results are affected by many variables and care must be taken in its application and interpretation.

Given these facts, caution should be exercised when performing Gram stains on liquid culture media. These media are designed, developed and manufactured for growing microorganisms. Therefore, liquid culture media should

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Newly Updated ­

BD Nasopharyngeal Specimen Collection Wall Chart

The BD Nasopharyngeal Specimen Collection Wall Chart has been updated and is now available. Utilize this reference by posting throughout your facility ­ wherever nasopharyngeal samples are collected. This chart is one of many tools that BD provides to customers free-of-charge. The BD Nasopharyngeal Specimen Collection Wall Chart provides guidance on methods of collection, materials and specimen transport to the laboratory. Four methods of collection are featured with illustrations and step-by-step instructions: · Vacuum-assisted nasopharyngeal aspirate collection, · Nasopharyngeal swab collection, · Nasopharyngeal wash ­ bulb collection, and · Nasopharyngeal wash ­ syringe collection. Also featured are BD CultureSwabTM collection devices and BD DirectigenTM rapid test kits that are appropriate for use with the nasopharyngeal collection methods. To obtain a copy of the BD Nasopharyngeal Specimen Collection Wall Chart for your facility, mark the appropriate box on the reader response card. If you have questions about BD respiratory collection and testing products, contact your local BD sales representative or call Technical Services at 800.638.8663.

Get Clued to the Flu

In response to the vaccine shortage, BD developed a public awareness campaign for use during this year's flu season: Get Clued to the Flu. The program emphasized the continuum of care and consisted of a four-part message: Get Informed ­ Get Dosed ­ Get Diagnosed ­ and Get Well. Each message outlined options for helping to reduce the impact of influenza. Fortunately, the 2004-2005 flu season got off to a slow start and did not peak until week 7 (Feb. 13-19, 2005). As a result, vaccine supplies were replenished and the CDC issued the Revised Interim Guidance for Late-Season Influenza Vaccination program in an aggressive effort to continue to vaccinate people in priority groups.

For more information on the 2004-2005 flu season, visit www.cdc.gov/flu/weekly/. For more information on the BD Flu Awareness Program, visit www.bd.com/flu/.

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Launching ­

Our Newly Reconfigured Web Site

We are very pleased to announce we are launching a newly reconfigured web site: www.bd.com/ds. Our goal in reconfiguring this web site was to create a user-friendly site where customers can access all relevant product information.

The first button on the navigation tool bar is Product Center. Pressing this button will guide you to our menu of products by category:

Collection and Transport Dehydrated Culture Media Prepared Media Environmental Systems Direct Testing/Serology Stains and Reagents Identification/Susceptibility Blood Culture Mycobacteria Testing Molecular Diagnostics Equipment and Supplies

Upon clicking on a product category, you will have access to the full range of products for that category. Then it's just a matter of drilling down to your specific product of interest. Once you have selected a product, a Product Summary page will appear and a Related Documents toolbar listing all of the technical information for that product including the Product Insert, Material Safety Data Sheet (MSDS), Certificate of Analysis (COA) and much more. In other words, all of the technical information available will be at your fingertips to download, print or e-mail as you wish. Two other important buttons on the navigation tool bar are Technical Center and Learning Center. At the Technical Center, you will find Documents, Services, Contact Information and Literature Requests. At the Learning Center, you will find Newsletters, Events, Biodefense, Training Schedule, ASM Presentations and Webinars. We hope you find our new web site useful. Happy surfing!

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BD Diagnostics Service Organization News

2005 Customer Training Calendar

The Technology Training Center (TTC) is pleased to announce the publication of its new 2005 Customer Training Calendar. The TTC is responsible for training customers on the use of BD instrumentation and software. We do this through regularly scheduled Training Classes at the Center in Sparks, MD. When you purchase a new BD PhoenixTM, BD ProbeTecTM, BD ViperTM or BD EpiCenterTM system, you will be scheduled to attend the corresponding training class at our facility outside of Baltimore. Your training includes

The Industrial and Clinical Media Advisory Team

The Industrial and Clinical Media Advisory Team (ICMAT), an entity of the BD Diagnostics Service Organization, includes members of the Technical and Informatics Services (TAIS) group and the Technology Training Center (TTC). The team is focused on supporting the DifcoTM and BBLTM brands of dehydrated and prepared media manufactured by BD Diagnostics. This team, consisting of highly motivated and skilled microbiologists who are experienced in the fields of both clinical and industrial microbiology procedures, is designed to provide our customers consultation and solutions to their media needs. The team is responsible for: · Providing clinical and industrial microbiology technical assistance for our media customers via our toll-free number 800.638.8663. · Championing technical and service issues that impact our worldwide media customers. · Monitoring customer product incident reports for media product lines and working closely with our Quality Department to communicate product trends. · Providing technical support tools through our Worldwide Service Organization to international customers. The Industrial and Clinical Media Advisory Team is committed to continuing the tradition of excellence of BD Difco and BBL media.

instruction on instrument and system operations, basic troubleshooting and recommendations for optimizing workflow in your laboratory. Sufficient time is allowed for you to develop comfort in the operation of instrumentation and using system procedures. In addition to offering application training, BD Service Engineering also offers training for Biomedical Engineers on such platforms as: BACTECTM 9000 Series and BACTECTM MGITTM 960 instruments. The Biomedical Engineering class schedule is also listed in our new calendar. All application training classes offered by the TTC are eligible for P.A.C.E.TM (ASCLS), and the California and Florida State Departments of Health Continuing Education Credits. To obtain your copy of the 2005 Customer Training Calendar, mark the appropriate box on the reader response card. We hope to see many of you in Baltimore soon!

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Micro Happenings

FIND and BD Combine International Efforts to Improve Rapid Tuberculosis Diagnosis for HIV-positive Patients in Developing Countries

FIND (Foundation for Innovative New Diagnostics) and BD have formed an international collaboration aimed at improving diagnosis of pulmonary tuberculosis (TB) in HIVinfected patients in developing countries. TB is the leading cause of death in AIDS patients in high-burdened countries, mainly in sub-Saharan Africa. TB is particularly difficult to diagnose in AIDS patients because they often have few or no TB bacteria in their sputum; thus, the standard diagnostic procedure using microscopy is insensitive. Classical culture methods for TB are more sensitive, but notoriously slow, typically requiring 21 to 42 days. BD has developed an improved culture method, the BD MGITTM (Mycobacteria Growth Indicator Tube) system, which provides results within 10 to 14 days. The BD MGIT technology can be used both for detection of the bacteria causing TB as well as for the determination of resistance to the TB drugs routinely used for treatment. The BD MGIT system is now widely used in industrialized countries but not in the developing world. Ed Ludwig, President and CEO, BD, said, "BD is committed to providing technologies which can help alleviate the impact of TB and HIV globally. BD is pleased to enter into this agreement with FIND, which is aimed at making these technologies more accessible to the public health sector of high-burdened countries." "Access to quality diagnostic equipment in countries with limited health resources will improve the management of tuberculosis in HIV-positive patients," said Dr. Mario Raviglione, Director of the World Health Organization (WHO) Stop TB Department. "This new agreement provides a blueprint for modern TB technology to be made more widely available globally, which will help reduce TB deaths and decrease transmission rates in high risk areas." The agreement between FIND and BD will operate in two stages. cooperation with WHO, the relevant working groups of the Stop TB Partnership, and the Consortium to Respond Effectively to the AIDS/TB Epidemic (CREATE) based at the Johns Hopkins Center for TB Research. The goal of these demonstration projects is to promote the use of this technology by national TB control and AIDS programs. According to Dr. G. Roscigno, CEO of FIND, "The collaboration with BD in the MGIT project is a significant and innovative step forward for FIND. FIND's demonstration projects using the BD MGIT technology will pave the way for the introduction of critical technologies in the public health sector of developing countries while contributing to a reduction in mortality particularly in HIV/ TB patients."

Implementation

The second stage is focused on sustainable implementation of this advanced diagnostic technology in the public health sector. Under this agreement, BD has committed to provide certain equipment, reagents, training and support to the public health sector in the high-burdened countries on terms that will enable them to be purchased and implemented on a sustainable basis. Helene Gayle, President of the International AIDS Society (IAS) and Director, HIV, TB and Reproductive Health Program at the Bill and Melinda Gates Foundation said, "We are particularly pleased that this innovative collaboration between BD and FIND is addressing one of the most urgent needs of the TB and HIV community in the developing world."

About FIND

FIND is the only non-profit organization dedicated solely to the development of diagnostic tests for infectious diseases in developing countries. FIND was established with funding from the Bill and Melinda Gates Foundation. Driven by the huge burden of disease, the existence of global control strategies and the capacity to treat detected cases, FIND has selected tuberculosis as an initial disease focus.

Demonstration Projects

First, FIND will perform and support projects using certain BD equipment, reagents, training and support to be purchased from BD in an effort to demonstrate the effectiveness of more rapid and accurate TB diagnosis in settings where TB and HIV are common and in areas with high prevalence of multiple-drug-resistant TB. FIND will conduct these projects in close

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Micro Happenings

105th ASM General Meeting in Atlanta, Georgia

Join us in Atlanta, June 5-9, for the 105th ASM General Meeting. BD Diagnostics Booth #1031 will provide a full line of diagnostic solutions for infectious disease management, ranging from effective sample collection to molecular-based pathogen detection and automated systems to determine appropriate antimicrobial therapy.

In addition, we will be featuring our new product innovations including: · BBLTM CHROMagarTM MRSA, for detection of nasal colonization by methicillin-resistant Staphylococcus aureus (MRSA) · BD CultureSwabTM MaxV specimen collection and transport swabs, for improved recovery of fastidious organisms such as Neisseria gonorrhoeae and Haemophilus influenzae · BD DirectigenTM EZ Strep A test and BD ChekTM Strep A test, utilizing lateral flow immunoassay technology to provide high accuracy of detection of betahemolytic group A Streptococcus

We will also feature the BD PhoenixTM Automated Identification/Susceptibility System, the BD ProbeTecTM ET DNA Amplification System and the BD ViperTM System, as well as other products. BD Diagnostics has the solutions for improving your patients' outcomes.

Foodborne Diseases

Continued from page 2

ciently pasteurized fluid milk, cheeses (particularly soft-ripened varieties), ice cream, raw vegetables, raw and cooked poultry, all types of raw meats and raw and smoked fish. The USDA

LabOTM is published three times per year by BD Diagnostics, 7 Loveton Circle, Sparks, MD 21152, 410-316-4701. Editor: Mary Jo Zimbro, B.S., MT(ASCP). Send address changes and mailing list additions to the attention of Marketing Communications, Mail Code 634. For technical information, call Technical Services, toll free, at 800-638-8663. Visit our web site at http://www.bd.com/ds/.

rated non-heated hot dogs as having the highest risk of Listeria contamination of the 20 categories of foods tested. When hot dogs are reheated, the risk for Listeria infection drops to one of the lowest among the foods tested. However, up to 14% of consumers eat hot dogs without first reheating them.4 L. monocytogenes is responsible for initiating a number of disease scenarios including septicemia, meningitis, encephalitis and intrauterine or cervical infections in pregnant women. Although all of the manifestations are serious, it is the last two that are of major concern since they may result in spontaneous abortion (during second/third trimesters) or stillbirth of the fetus. Also, various reports have noted that gastrointestinal symptoms may be the preceding signs to the more serious forms of listeriosis or may be the only symptoms being expressed. The twist here is that gastrointestinal symptoms are not commonly associated with ingestion of Listeria contaminated foods.7 It is suspected that the incubation period for the more serious manifestations may range from 48-72 hours to a few weeks, whereas, the gastrointestinal incubation period is believed to be 12 hours. The bottom

line is that the pathogenesis of L. monocytogenes is its ability to proliferate unchecked inside phagocytic host cells.7 A varying transit carrier state of 2 to 20% exists in both humans and animals.5 The antimicrobial susceptibility patterns for treating listeriosis have remained relatively stable for many years. Although various antibiotics have been used for treatment, penicillin or ampicillin, with or without an aminoglycoside, is recommended for treatment. The antibiotic administered should reach a high concentration in phagocytic cells, be able to cross the blood-brain barrier and have rapid bactericidal activity.5

Centers for Disease Control and Prevention. 2001. Morbid. Mortal. Weekly Rep. 50(13): 241. 2 Food Quality Magazine. 1998. June/July: 33-36. 3 Stern and Line. 2001. The microbiological safety and quality of food, V.II. 1043, 1178, Aspen Pub., Gaithersburg, MD. 4 "Food Safety Net". April 11, 2001 (newsletter), www.foodsafetynetwork.ca/ 5 Bill and Doyle. 1991. In Balows, Hausler, Herrmann, Isenberg and Shadomy (ed.), Manual of clinical microbiology, 5th ed., ASM, Washington, D.C. 6 "Bad Bug Book", re. May 4, 2001, www.cfsan.FDA.gov. 7 Benfield-Capers et al. 2003. APHL Food Safety Laboratory Capacity Assessment Project, 1-37 (Special Report).

1

CHROMagar is a trademark of Dr. A. Rambach. Difco is a trademark of Difco Laboratories, subsidiary of Becton, Dickinson and Company. P.A.C.E. is a trademark of The American Society for Clinical Laboratory Science. Unless otherwise noted, BD, BD Logo and all other trademarks are the property of Becton, Dickinson and Company. BD Diagnostics is an ISO 9000 registered manufacturer. ©2005 Becton, Dickinson and Company 0-2759 Printed in USA

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