Subject: Microbiology Topic: Bacteria Associated with Skin

Infection 2

Lecturer: Dr. Eleanor Padla Date of Lecture: Transcriptionist: Broken, Blue Blink Editors: Bluebolt, Aquamarine Pages: 9

Bacteria Associated with Skin Infection Erysipelothrix rhusiopathiae General Properties: straight/ slightly curved Gram (+) rods, in singles, short chains, V/L configuration ubiquitous in nature Determinants of Pathogenicity: 1. Hyaluronidase hydrolyzes hyaluronic acids in connective tissues

2. Neuraminidase Cleaves N-acetylneuraminic acid

Transmission: follows subcutaneous inoculation of the organism swine, fish and/or their products most often associated with human disease occupational exposure Clinical Manifestations of Erysipelothrix rhusiopathiae 1. Erysipeloid a) non-suppurative, purplish erythematous lesions at site of inoculation b) lesions burn and itch with with

2. diffuse cutaneous eruption systemic symptoms 3. bacteremia endocarditis associated

SY 2011-2012

Prevention: protection from direct contact with infected animal secretion live attenuated vaccine (for veterinary use)

Laboratory Diagnosis 1. Gram Stain Gram (+) ----------------------------------------------------------------------------------------------------------------------------Question and Answer Portion: 1. Q: How does Erysipelothrix rhusophiatiae appear after Gram staining? A: Gram (+) bacilli 2. Q: Is Erysipelothrix rhusiopathiae a spore-forming organism? A: NO

3. Q: What is the major portal of entry of Erysipelothrix rhusiopathiae ? 2. Culture a. specimens biopsies from lesion b. media i. ii. c. growth CO2 HI broth, BAP -hemolytic improved by 5-10 5. Q: What is the enzyme produced by virulent strains of E. rhusiopathiae that cleaves sialic acid on cell surfaces? A: Neuraminidase aspirates/ margin of A: Skin 4. Q: Does Erysipelothrix rhusiopathiae cause erysipelas? A: NO. Erysipelas is a superficial bacterial skin infection that characteristically extends into the cutaneous lymphatics. It is caused by Strep pyogenes. Erysipelothrix rhusiopathiae cause erypiseloid

d. produces H2S

3. Mouse Protection Test a. Treatment: Penicillin to confirm isolates

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BACILLUS General Properties: large, Gram (+) spore-forming rods, in chains spores oval, centrally located usually motile form rhizoid colonies

straight rods with truncated ends, jointed bamboo-rod appearance (fig below) spores formed in dead/dying animals oval, central, do not distend the cell capsulated non-motile culture, by

*Bacillus is a gram-positive bacteria, I dont know why it appeared dark pink during Dras lecture. Maybe it has something to do with the contrast or picture properties.

Determinants of Pathogenicity: I. capsular polypeptide glutamic acid of D-

plasmid-encoded (pXO2) anti-phagocytic detected by Mac Fadyean reaction (polychrome methylene blue) II. Anthrax toxin o Clinically Important Species 1. Bacillus anthracis anthrax major virulence factor o heat-labile

o plasmid-encoded (pXO1) o anthrax consists of: toxin is

2. Bacilus cereus food poisoning ocular infections

protective antigen (PA) edema factor (EA) Lethal factor (LA) synergistically

systemic infections o 1. B. anthracis General Properties:

produce edema

cell

death

and

Pls refer to the last page for clearer and bigger image. Pls know this by heart <3

Characteristic black eschar surrounded by a ring of vesiculation. There is much edema of the face and neck. *** An eschar is a slough or piece of dead tissue that is cast off from the surface of the skin.

Anthrax: disease of herbivores human cases: agricultural industrial

Clinical manifestations cutaneous pustule) pulmonary disease) gastrointestinal anthrax) (malignant

(Woolsorters

(bowel

Eight days later, the eschar has enlarged to cover the previously vesicular area, and the surrounding edema has diminished.

Epidemiology

Above picture Reaction

shows

MacFadyean

Above picture is stained using India ink. Notice the white capsule of the B. anthracis. 3. Culture and Identification Pls refer to the last page too Know it by <3 as well. a. specimens - material from pustule b. media Laboratory Diagnosis 1. Gram stain gram (+) i. ii. rhizoid 4. Guinea Pig Lethality Test 5. Serologic Tests BAP non-hemolytic,

2. Capsule stain

4. Q: What is the infective particle of B. anthracis ?

A: spores

5. Q: Is anthrax directly transmissible from one infected animal or person to another?

Treatment: penicillin

Prevention: control of disease in animals careful handling animals/ products vaccination killed spore HUMAN vaccine for of infected

A: NO. Transmission of anthrax is transferred indirectly. Transmission in humans is acquired by the entry of spores through injured skin (cutaneous anthrax) or rarely by mucuous membranes (gastrointestinal anthrax), or by inhalation of spores into the lungs (inhalation anthrax). ----------------------------------------------------------------------------------------------

living spore vaccine (Stearne strain) for ANIMALS ----------------------------------------------------------------------------------------------------------------------------Question and Answer Portion again haha 1. Q: Is Bacillus anthracis motile nor non-motile? A: Non-motile 2. Q: What is the major component of B. anthracis capsule? Anaerobes unable to utilize O2 as final electron acceptor lack cytochrome lack catalase lack superoxide dismutase generate energy solely by fermentation growth requirements: A: D-glutamic acid low oxygen tension ( 10% PO2) 3. Q: Anthrax toxin is composed of how many protein components? A: 3; namely protective antigen (PA); edema factor (EA); Lethal factor (LA) PA + EA or PA + LA= Toxicity PA + EA + LA= Full Toxicity (causing edema and cell death) reduced ORP (Eh) 5% CO2

enriched medium Features Associated With Anaerobic Infections

often

caused

by

endogenous

opportunistic pathogens usually not transmissible occur in settings of compromised host defense usually polymicrobic abscess necrosis formation and tissue

widely distributed in nature spores produced anaerobically, usually distended cells most species motile medically important species: C. perfringens - wound & soft tissue infections, food poisoning, enteritis necroticans, septicemia C. tetani - tetanus C. botulinum botulism C. difficile Antibiotic Associated Diarrhea, Pseudomembranous Collitis Clostridium perfringens (formerly known as C. welchii) General Properties: short, plump Gram (+) rods with squarish ends aerotolerant non-motile, capsulate spores oval, central, do not distend cell; rarely observed part of Normal Flora of GIT, female genital tracts 5 types of C. perfringens (A- E) = type A, C most common seen in human diseases

infections developing slowly, many become chronic putrid odor of infected material/ culture gas in tissues/ loculations (-) aerobic cultures

Laboratory Diagnosis: 1. Gram stain 2. Culture & Identification a. Putrified Specimens i. ii. iii. aspirates iv. urine (suprapubic, catheterized) abscess contents, deep wound aspirates blood, fluids CSF, body

transtracheal

b. Media- EYA, chopped meat medium, LKVB 3. culture systems anaerobic chamber Prevention: avoid conditions that reduce ORP in tissues prevent introduction of anaerobes (from NF) into wounds, etc protect against toxin Gaspak;

Determinants of Pathogenicity at least 12 soluble substances produced;

4 are major lethal toxins a toxin - produced by all types of C. perfringens b toxin - produced by types B, C e toxin - produced by types B, D

CLOSTRIDIA

i (iota) toxin - produced by 1

type E toxin lecithinase C (phospholipase C) causes disruption/ leakage of cell membranes

Laboratory Self explanatory naman to Wound and Soft Tissue Infections: simple wound contamination crepitant cellulitis clostridial myonecrosis/ gas gangrene Clostridial gangrene): myonecrosis (gas 1.

Diagnosis:

Gram Stain predominance of Gram (+) rods, sparse or no wbc spores rare in materials, cultures clinical

highly lethal, necrotizing soft tissue infection of skeletal muscle myonecrosis, muscle swelling, severe pain, gas production, sepsis may also be caused by other clostridial species disease process includes tissue inoculation, low oxygen environment

Hemolysis on BAP Naggler Reaction - a test for the identification of alpha toxin of Clostridium perfringens; the addition of antitoxin to cultures on egg yolk agar prevents visible opacity, due to lecithinase action which is

normally observed around colonies.

A: Clostridium welchii 2. Q: Is Clostridium perfringens motile nor non-motile?

A: Non-motile Reverse CAMP The test organism, Clostridium perfringens, is streaked down the center of the plate. Streptococcus agalactiae (group B) is streaked at right angles to the Clostridium perfringens. A positive reverse CAMP result for Clostridium perfringens is shown by the arrow-shaped zone of enhanced hemolysis pointing towards Clostridium perfringens. 3. Q: What is the principal toxin of C. perfringens ? A: alpha-toxin/ lecithinase C (phospholipase) 4. Q: What is the most frequent source of C. perfringens ? A: GIT 1. Q: What is the hemolytic pattern of C. perfringens colonies when grown anaerobically on BAP? A: beta-hemolysis CLINICAL CORRELATIONS CASE # 1 A 69-year-old male, employed as a butcher, was seen in the dermatology clinic for a localized lesion on the index finger that was associated with pain and swelling. The lesion was well defined and slightly elevated, with a peripheral zone of discoloration, but was not edematous. A tissue sample was obtained and submitted to the laboratory for Gram stain and culture. Gram stain revealed moderate PMN but no organisms. E. rhusiopathiae was isolated from the thioglycollate broth on culture day 7. Treatment with a topical bacitracin ointment resulted in resolution of symptoms. 1. Erysipelothrix rhusiopathiae is: A. Gram (+) aerobic cocci B. Gram (+) anaerobic bacilli C. Gram (+) sporulating bacilli D. Gram (+) non-sporulating bacilli 2. Diagnosis of E. rhusiopathiae infection is confirmed following isolation of the organism from? A. feces B. tissue biopsy C. Both A and B

Stormy fermentation in milk: In anaerobically-grown Litmus Milk cultures, enzymes of C. perfringens ferment lactose to acid and gas which influence caesin precipitation forming a curd-like mass and its distortion due to the gas. This is "stormy fermentation. Treatment: cleansing/ surgical management of necrotic tissue antibiotic therapy hyperbaric oxygen antitoxin ( for histotoxic clostridia) -----------------------------------------------------------------------------------------------------------------------------

Question and Answer again and again 1. Q: What is the former name of C. perfringens ?

D. Neither A nor B CASE # 2 A 56-yo male postal worker from Washington DC, presented to the ER. He had been well until 3 days PTA, when he developed low-grade fever, chills, cough, dypsnea on exertion, and generalized malaise. Sputum was initially clear until the night of admission, when it became blood-tinged. Blood cultures showed prominent Gram-positive rods consistent with B. anthracis. 1. Which is/are true of B. anthracis? A. used as biologoical warfare B. motile, non-capsulated organism C. forms spore under anaerobic conditions D. all of the above

CASE # 3 A 30-yo man presented at the DLSUMC with severe pain in the right calf. Three days earlier, he required suturing of a deep laceration to his calf sustained in a car accident. Over the past 48 hrs, his right calf was edematous with deep erythema surrounding the suture line. Hemorrhagic bullae and brownish exudates from the wound were noted. He also experienced intermittent low grade fever. Ho has not taken any medications. Wound exudate Gram stain showed few WBCs and abundant Gram (+) rods. 1. The most likely causative agent is A. S. pyogenes B. C. perfringes C. B. anthracis D. C. tetani 2. Which is the mainstay of therapy in this case? A. Surgical debridement B. Hyperbaric oxygen therapy C. Tetanus toxoid immunization D. Immunization with polyvalent antitoxin Answers: 1. D; B 2. A; B; A 3. B; A End of transcription
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2. Which is/are impt factor(s) if B. anthracis? A. Spore B. 3 component toxins C. Both a and b D. Neither a nor

virulence

3. Human cases of anthrax reflects the organisms:

A. Mode of entry B. Tissue tropism C. Host specificity D. Invasive properties

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