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Bacterial Information and Courses from MediaLab, Inc.

These are the MediaLab courses that cover Bacterial and links to relevant pages within the course.

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Laboratories Individuals

Cerebrospinal Fluid
The cells present in this field may indicate what condition?View Page
Increased numbers of the cells shown at the right are indicative of what condition?View Page
Which of the following statements is true about the type of cell shown at the right?View Page
Table of Conditions

The following table lists the various cell types and macroscopic descriptions of CSF, and the patient conditions that could cause those properties to be present in the patient's CSF: Predominant Cell Appearance Conditions lymphs variable; clear - turbid viral meningitis tubercular meningitis multiple sclerosis drug abuse lymphoma leukemia Guillain-Barré syndrome chronic alcoholism neutrophils variable; clear - turbid bacterial meningitis mycotic meningitis early tuberculosis hemorrhage cerebral abscess tumors monocytes variable chronic bacterial meningitis partial treatment of meningitis tumors macrophages clear - turbid or clear - xanthochromic bloody tuberculosis fungal meningitis following hemorrhage blood contamination eosinophils variable parasitic meningitis fungal meningitis allergic reaction medications shunts dyes tumor cells variable metastatic carcinoma blast cells variable leukemia lymphoma normal to increased lymphs clear - xanthochromic benign tumor spinal cord brain ependymal or orchoid cells (often clumped) variable; may be xanthochromic bloody trauma spinal tap Adapted from Saunders Manual of Clinical Laboratory Science. Craig A. Lehrmann, Ed. WB Saunders, 1998.

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Mature Peripheral Blood Cells

In normal spinal fluid from an adult, 60% of cells are lymphocytes and up to 30% are monocytes. Neutrophils abundance up to 2% is also considered within normal limits when a cytospin smear is used for the differential. In children, normal CSF cells are 70% monocytes, up to 20% lymphocytes and up to 4% neutrophils. When any of these normal cell abundances are increased, the term pleocytosis is used. Neutrophil pleocytosis is an increase in neutrophils and usually indicates the presence of a bacterial infection.

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Match the condition on the left with associated cells on the right.View Page
Monocytes (continued)

Monocytes are also present in this field. Increased numbers of monocytes may be seen in chronic bacterial meningitis, multiple sclerosis, fungal meningitis, CNS malignant disorders or CNS hemorrhage.

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Lymphocytes

Many lymphocytes are present in this field. Two larger, atypical lymphocytes with intact cytoplasm and slightly indented nuclei are seen near the center of this slide. Two other large cells with irregular, trailing cytoplasm are macrophages (histiocytes). Increased lymphocytes may be seen in viral meningoencephalitis, partially treated bacterial meningitis, multiple sclerosis, Guillian-Barre's syndrome, or polyneuritis.

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Neutrophils

Many neutrophils are present in this slide from a spinal fluid sample from a patient with bacterial meningitis. Several macrophages (histiocytes) which have engulfed some of the bacteria can also be seen among the neutrophils.

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Chemical Screening of Urine by Reagent Strip
False-positive tests for protein may be caused by: (Choose ALL of the correct answers)View Page
What affect may bacterial contamination have on urine pH?View Page
A voided urine specimen is delivered from the women's clinic to the laboratory six hours after collection. The following results are reported: Color: yellowProtein: negativeBilirubin: negative Clarity: cloudyGlucose: negativeUrobilinogen: 0.2 mg/dL Sp. Gravity: 1.020Ketone: negativeNitrite: positive pH: 9.0Blood: negativeLeukocyte esterase: negativeWhat might these results indicate?View Page
Acid and alkaline urine pH

Reasons for acidic urine pH include: a high-meat diet, respiratory/metabolic acidosis, and hypochloridemia. A urine with a high concentration of glucose may also have a lower pH. An alkaline pH may be the result of a vegetarian diet, respiratory/metabolic alkalosis, or a bacterial infection caused by urease-producing bacteria. Urine that contains bacteria can become more alkaline if the specimen remains at room temperature for an extended period of time.

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False Positive Protein Results

A urine specimen that has remained at room temperature for an extended period of time may produce a false-positive protein result on a reagent strip. A false positive may also occur in the presence of bacterial contamination, alkaline medication, or quaternary ammonium compounds such as disinfectants or drugs, and with skin cleansers containing chlorhexidine.

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False Positive Results

A false positive nitrite test result may occur when a urine specimen has remained at room temperature for an extended period of time, allowing bacterial contaminants to multiply and produce measurable levels of nitrites. Interference from some medications that cause the urine to become red or orange may lead to an incorrect reading of positive for nitrite.

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Clinical Significance

Using the esterase test in conjunction with pH, protein and nitrite provides a combination of tests which can screen for the presence of bacterial infection.

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CLIA Blood Banking Review
Which of the following is most commonly associated with febrile non-hemolytic transfusion reactions:View Page

CLIA Chemistry / Urinalysis Review
The elements indicated by the arrows are more likely to be seen in patients with which condition:View Page
Elevation in CSF total protein may be seen in all of the following conditions except:View Page
Which of the following statement about synovial fluid is true:View Page
Bacterial contamination of a urine specimen from a normal healthy individual could originate from all of the following except:View Page
Detection of a fruity odor in a fresh urine sample may be indicative of:View Page

CLIA General Laboratory Review
C-reactive protein:View Page
A patient with atypical (reactive) lymphocytes in his peripheral blood smear should be tested for:View Page

CLIA Hematology / Hemostasis Review
The predominant cells seen on the CSF smear in this illustration are indicative of:View Page
The predominant cells seen in this CSF are suggestive of:View Page
The large number of these cells seen in the CSF cytocentrifuged smear in this illustration is suggestive of:View Page

CLIA Microbiology / Serology Review
The most frequent cause of bacterial meningitis in older adults is:View Page
With regard to blood cultures, which blood to broth ratio is most conducive to growth:View Page
The turbidity of the bacterial inoculum of a standard disc diffusion (Kirby-Bauer) susceptibility test is:View Page

Confirmatory and Secondary Urinalysis Screening Tests
A urine specimen to be tested for bilirubin arrives in the laboratory after sitting on the counter at the nurses station for 2 hours. Which of the following statements describes the Ictotest® reaction that could potentially occur in this situation?View Page

Current Topics in Clinical Microbiology
The name of the rapid test as illustrated in this photograph, often used to differentiate S. pneumoniae from viridans streptococci, in which a drop of 10% deoxycholate was placed on an area of growth, is:View Page
Review 2

Tuomanen EI.: Pathogenesis of pneumococcal inflammation: otitis media Vaccine. 19 Suppl 1:S38-40, 2000Pneumococci cause damage to the ear in otitis media with an association with bacterial meningitis. The pathogenesis of injury involves host response to cell wall constituents and the pore-forming toxin, pneumolysin.Release of cell wall constituents, particularly during antibiotic-induced bacterial lysis, leads to an influx of leukocytes and subsequent tissue injury. The signal transduction cascade for this response is becoming defined and includes CD14, Toll-like receptor 2, NFkB, and cytokine production.The second source of injury is the cytotoxicity of the pore forming toxin, pneumolysin.Decreasing the sequelae of otitis can be achieved by an increased understanding of the site-specific mechanisms of pneumococcal-induced inflammation.

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Middle ear damage in cases of S. pneumoniae infections are caused primarily by:View Page
The colonies shown in the blood agar (upper) and MacConkey agar (lower) biplate are a 24 hour growth from an aerobic blood culture bottle that became positive at 12 hours after inoculation. The appearance of the colonies on MacConkey agar rules out the following two bacterial species:View Page
The bacterial species shown growing on 5% sheep blood agar was recovered from the spun sediment of a midstream urine specimen after 24 hours incubation at 35C. Each of the following tests would be useful in supporting the presumptive identification of Enterococcus species except:View Page
The gram stain shown in the photograph was prepared from a positive anaerobic blood culture bottle after 36 hours incubation. Based on the morphology of the bacterial cells (some with spores--blue arrows), the most likely identification is:View Page
A gram stain of the serous exudate is shown in the photograph. The appropriate report would read:View Page
The bacterial cells shown in the photograph were observed in a smear prepared from the colony shown before. Which of the following tests will help to affirm the identification of Staphylococcus aureus?View Page
Review 1

Francois P. Vaudaux P. Foster TJ. Lew DP.: Host-bacteria interactions in foreign body infections. Infection Control & Hospital Epidemiology. 17:514-20, 1996Persistent staphylococcal infections are a major medical problem, especially when they occur on implanted materials or intravascular catheters.This review describes some of the recently discovered molecular mechanisms of Staphylococcus aureus attachment to host proteins coating biomedical implants.These interactions involve specific surface proteins, called bacterial adhesins, that recognize specific domains of host proteins deposited on indwelling devices, such as fibronectin, fibrinogen, or fibrin.Elucidation of molecular mechanisms of S. aureus adhesion to the different host proteins may lead to the development of specific inhibitors blocking attachment of S. aureus, which may decrease the risk of bacterial colonization of indwelling devices.

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Decreasing the risk of staphylococcal colonization of indwelling catheters in the future may involve:View Page
A Brown and Brenn gram stain was performed on one of the tissue biopsy specimens. Organisms were seen as shown in the photograph. Based on the history and the appearance of the bacteria, the most likely identification is:View Page
Listeria motility

Listeria monocytogenes is optimally motile at 25C; and is non-motile at 35-37C.Motility may be directly assessed by observing bacterial cells with a tumbling motion in a direct mount preparation.In soft motility agar, the identification can be made by observing for a thin, umbrella-like lateral extension of growth from the stab line (see photograph).

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The test(s) which may be performed to establish a presumptive differential identification between group B streptococci and L. monocytogenes is/are:View Page
Review 2

Low JC. Donachie W.: A review of Listeria monocytogenes and listeriosis. Veterinary Journal. 153:9-29, 1997Following the initial isolation and description in 1926, Listeria monocytogenes has been shown to be of world-wide prevalence and is associated with serious disease in a wide variety of animals, including man.Our knowledge of this bacterial pathogen and the various forms of listeriosis that it causes has until recently been extremely limited, but recent advances in taxonomy, isolation methods, bacterial typing, molecular biology and cell biology have extended our knowledge. It is an exquisitely adaptable environmental bacterium capable of existing both as an animal pathogen and plant saprophyte with a powerful array of regulated virulence factors.Most cases of listeriosis arise from the ingestion of contaminated food and in the UK the disease is particularly common in ruminants fed on silage.Although a number of forms of listeriosis are easily recognized, such as encephalitis, abortion and septicaemia, the epidemiological aspects and pathogenesis of infection in ruminants remain poorly understood. The invasion of peripheral nerve cells and rapid entry into the brain is postulated as a unique characteristic of its virulence, but relevant and practical disease models are still required to investigate this phenomenon.

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Review 2

Low JC. Donachie W.: A review of Listeria monocytogenes and listeriosis. Veterinary Journal. 153:9-29, 1997Following the initial isolation and description in 1926, Listeria monocytogenes has been shown to be of world-wide prevalence and is associated with serious disease in a wide variety of animals, including man.Our knowledge of this bacterial pathogen and the various forms of listeriosis that it causes has until recently been extremely limited, but recent advances in taxonomy, isolation methods, bacterial typing, molecular biology and cell biology have extended our knowledge. It is an exquisitely adaptable environmental bacterium capable of existing both as an animal pathogen and plant saprophyte with a powerful array of regulated virulence factors.Most cases of listeriosis arise from the ingestion of contaminated food and in the UK the disease is particularly common in ruminants fed on silage.Although a number of forms of listeriosis are easily recognized, such as encephalitis, abortion and septicaemia, the epidemiological aspects and pathogenesis of infection in ruminants remain poorly understood. The invasion of peripheral nerve cells and rapid entry into the brain is postulated as a unique characteristic of its virulence, but relevant and practical disease models are still required to investigate this phenomenon.

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Review 3

Rouquette C. Berche P. The pathogenesis of infection by Listeria monocytogenes Microbiologia. 12:245-58, 1996 Listeria monocytogenes is a Gram-positive bacterium responsible for severe infections in human and a large variety of animal species. It is a facultative intracellular pathogen which invades macrophages and most tissue cells of infected hosts where it can proliferate. The molecular basis of this intracellular parasitism has been to a large extent elucidated. The virulence factors, including internalin, listeriolysin O, phospholipases and a bacterial surface protein, ActA, are encoded by chromosomal genes organized in operons. Following internalisation into host cells, the bacteria escape from the phagosomal compartment and enter the cytoplasm. They then spread from cell to cell by a process involving actin polymerisation. In infected hosts, the bacteria cross the intestinal wall at Peyer's patches to invade the mesenteric lymph nodes and the blood. The main target organ is the liver, where the bacteria multiply inside hepatocytes. Early recruitment of polymorphonuclear cells lead to hepatocyte lysis, and thereby bacterial release This causes prolonged septicaemia, particularly in immunocompromised hosts, thus exposing the placenta and brain to infection. The prognosis of listeriosis depends on the severity of meningoencephalitis, due to the elective location of foci of infection in the brain stem (rhombencephalitis). Despite bactericidal antibiotic therapy, the overall mortality is still high (25 to 30%).

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A bacterial isolate that produces pitting of the agar and has a bleach-like odor is probably E. corrodens. Closely related species that must be ruled out include:View Page

Erythrocyte Inclusions - Wright Stained Smears
Coarse basophilic stippling is usually seen in patients who have:View Page

HIPAA Privacy and Security Regulations
Case Study: De-identified Health Information. You work in a laboratory microbiology department which provides a local nursing home with information about the effectiveness of various antibiotics it uses to treat infections. You print the requested information, including complete patient identification, bacterial organisms identified, and their sensitivity to various antibiotics. What information should you provide to the nursing home?View Page

Phlebotomy
Introduction

Blood is normally sterile. Any bacterial growth in the bloodstream is abnormal, and is an important cause of fever.Blood culture means the incubation of blood in appropriate media to allow growth and identification of bacteria or other organisms that may be present in a patient’s bloodstream. Blood cultures are performed on febrile patients to identify and treat bloodborne organisms with the most appropriate antibiotic.

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Reading Gram Stained Direct Smears
Identification of bacteria

Identification of bacteria in direct smears may be of lifesaving importance. For example, a rapid diagnosis of bacterial meningitis, made after examining a gram stained smear of the patient's cerebrospinal fluid, allows the physician to begin treatment immediately.The appearance of bacteria on gram stained smears is suggestive of a certain species, but identification may not be made on the basis of the stain alone. An exception to this rule is the presence of gram negative intracellular diplococci from a male urogenital specimen, which is presumptive identification of Niesseria gonorrhoeae.In addition, culture results can be correlated with the direct smear report.

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Recording Bacterial Characteristics

The Gram stain reaction, shape, and arrangement of bacteria, and the presence or absence of intracellular organisms must be noted on the worksheet.Examples:Gram positive cocci in chains are present.Gram negative diplococci, intracellular, are present within white blood cells.Quantitate by approximating the average number of each cell type seen in 10 oil immersion fields, and record as:Many = More than 15/fieldModerate = 4-15/fieldFew = 1-3/fieldOccasional = 2-10/10 fieldsRare = 0-2/10 fields

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Significance of Specific Findings:

Epithelial cells in large numbers within sputum smears means that the specimen is predominantly oral saliva, rather than true sputum from the lung. Epithelial cells in urine smears indicate that the sample has been contaminated by organisms found on the vulva or distal urethra. Bacteria found near or on epithelial cells are usually normal contaminating bacterial flora.White blood cells indicate inflammation and possible infection. The direct smear examination should focus within and around these cells.Red blood cells in a direct smear are not usually significant.Yeast may be present as normal flora in upper respiratory tract or genital tract. They may be significant if they predominate, or if budding yeast forms are seen.Hyphae are more likely to indicate the presence of fungal infection, but this determination requires correlation with clinical findings.Bacteria found in spinal fluid, blood, tissue and specimens from other sterile sites are always significant.Body fluids which are normally sterile must be examined carefully. If only one organism per oil immersion field is identified, then there are about 105 organisms per mL present in the sample! Bacteria observed in specimens from the throat, genital tract and other areas containing normal flora suggest infection only if their composition and type varies significantly from the norm.

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Reporting Direct Smear Results

Direct smear results are generally reported in the same way that they are read, except that bacterial cell arrangement (ex: clusters, chains, pairs) may be misleading and is generally not reported except in the case of intracellular diplocci in genital smears.

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Reading Gram Stained Smears From Cultures
The arrangement of bacterial cells is noted but not usually reported.View Page
Bacterial arrangements that can be seen on a smear include:View Page
The process of microorganism identification includes: (Choose ALL of the correct answers)View Page
Culture, Isolation, and Identification of Microorganisms

The process of culture, isolation and identification of microorganisms is basic to medical microbiology. When a culture shows signs of growth, the process of identification includes examining the following characteristics:appearance of the colonies in the culture mediumstaining reactionappearance of stained organismssizeshapearrangement of bacterial cellsThis type of preliminary identification may help the physician to initiate the appropriate antibiotic treatment.

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Red Cell Disorders: Peripheral Blood Clues to Nonneoplastic Conditions
Conditions suggested by the macrocytes and the neutrophil in the photograph to the right include:View Page

Variations in White Cell Morphology - Granulocytes
Cytoplasmic Vacuolation

Vacuoles are areas of the cytoplasm which do not stain with Wright's stain and appear as holes in the cytoplasm. Their composition may vary; some will contain remnants of bacterial digestion, autodigestion in an aging cell, while still others may contain fat. It is not possible to differentiate the various types of vacuoles on Wright's stained smears under light microscopy. They may be seen occasionally in an aging granulocyte, but are seen more frequently and are significant in cases of bacterial infection and septicemia.

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A vacuole may contain:View Page
Toxic granulation is seen most frequently in:View Page

White Cell and Platelet Disorders: Peripheral Blood Clues to Nonneoplastic Conditions
The peripheral blood smear tagged in the photograph was held for review because of too many platelets, about double the normal average of 8 - 15/oil immersion field or one per 10 - 20 RBC's. Conditions in which platelets are increased as noted in the photograph include:View Page


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