Liam - GPC Strep, GPC Staph, Clostridium,Bacilli,Non-ES forming GPR

Chapter 19: Pathogenic Gram Positive Bacteria

General Information on Genus Streptococcus
  • Characteristics:

    • Gram-positive cocci found in pairs or chains.

    • Catalase negative (do not produce the enzyme catalase).

    • Categorized by hemolysis type and Lancefield classification.

    • Some strains require CO2 for enhanced growth.

    • Typically require blood for cultivation.

Hemolysis Types in Streptococci
  • Beta (β) Hemolysis:

    • Complete lysis of red blood cells (RBCs).

    • Exhibits a clear zone around colonies (e.g., Group A Streptococcus).

  • Alpha (α) Hemolysis:

    • Partial lysis of RBCs.

    • Exhibits a green zone around colonies (e.g., Streptococcus pneumoniae).

  • Gamma (γ) Hemolysis:

    • No lysis of RBCs.

    • No hemolytic activity (e.g., Enterococcus sp.).

Lancefield Classification
  • Developed by Rebecca Lancefield based on bacterial cell wall antigens.

  • Major Groups:

    • Group A: Streptococcus pyogenes (S. pyogenes)

    • Group B: Streptococcus agalactiae (S. agalactiae)

    • Group D: S. bovis

    • Groups C, F, G: Other less common species.

Diseases Associated with Streptococcus pyogenes
  • Pharyngitis (Strep Throat):

    • Can progress to Scarlet Fever characterized by red cheeks and rash due to erythrogenic toxin.

  • Impetigo/Pyoderma:

    • Highly infectious skin condition.

  • Necrotizing Fasciitis:

    • Known as "flesh-eating strep"; extremely severe with up to 50% fatality rate.

  • Consequences of Untreated Infections:

    • Autoimmune response leading to Rheumatic Heart Fever (RHF) or Glomerulonephritis.

Diagnostic Tests for Streptococcus
  • Rapid Strep Test: Up to 30% false negatives.

  • Bacitracin Test: Reference for identifying Group A Strep.

  • Latex Agglutination Test: For laboratory identification of Streptococcus species.

Treatment of Beta-Hemolytic Group A Infections
  • Remain sensitive to Penicillin.

  • Alternatives for penicillin-allergic patients include cephalosporin or erythromycin.

  • Bacitracin is effective topically.

  • Necrotizing fasciitis requires surgery.

Streptococcus agalactiae (Group B)
  • Primarily affects newborns.

  • Colonizes in GI, urinary, and genital tracts.

  • Disease risk is higher if maternal antibodies are absent:

    • Early-onset: Infects newborns during birth.

    • Late-onset: Infection from caretakers after the first week of life.

  • Can cause severe conditions like sepsis, meningitis, and pneumonia.

Prevention of Group B Streptococcus Disease
  • Expectant mothers are cultured for BSGB before birth.

  • If positive, IV antibiotics (such as Penicillin G) are administered during labor.

  • This practice has reduced early-onset infections by 70%.

  • A BSGB vaccine is in development.

Identifying Group B Streptococcus
  • Characterized by beta hemolysis.

  • CAMP Test: Enhances the beta hemolysis of Staphylococcus aureus.

  • Antigen detection tests from culture samples.

Non-Beta-Hemolytic Streptococci
  • Streptococcus pneumoniae:

    • Diplococci appearance (lancet-shaped).

    • Major cause of pneumonia and other infections.

  • Viridans Streptococci (e.g., S. mutans):

    • Associated with dental caries and endocarditis.

  • Streptococcus bovis:

    • Found in the GI tract and associated with colon cancer.

  • Enterococcus:

    • Thrives in the intestinal tract, resistant to various conditions, indicator of fecal contamination.

Treatment of Enterococcal Diseases
  • Commonly found in UTIs, lungs, and causing septicemia.

  • Accounts for 10% of nosocomial infections.

  • Increased resistance, such as Vancomycin-resistant Enterococcus (VRE).

Overview of Acid-Fast Bacteria

  • Definition: Acid-fast bacteria are those that retain the primary stain even when treated with a strong acid alcohol, indicating the presence of mycolic acid in their cell walls.

  • Key Staining Characteristics:

    • Special Stain: Requires an acid-fast stain to rinse primary stain off, which is different from the Gram stain process.

    • Result: If a bacterium is acid-fast, it will not wash out the primary stain.

  • Common Acid-Fast Bacteria: Includes species from Mycobacterium, Nocardia, and Actinomycetes.

Structure of Acid-Fast Bacterial Cell Walls

  • Cell Wall Composition:

    • Thick layer of Mycolic Acid above peptidoglycan.

    • Presence of Arabinogalactan linking mycolic acids and peptidoglycan.

  • Permeability:

    • Small hydrophilic molecules require porins to pass through the outer membrane (10 nm) adjacent to a cell membrane (7 nm).

Mycobacterium Species Overview (True Acid-Fast Bact)

  • Characteristics:

    • Waxy, water-resistant cell walls due to lipid mycolic acids-

    • Mycolic acivd keeps bact from drying out, increases resistance to AB’s and Chem DI’s but also slows down growth, nutrients dont enter as easily.

    • Grows intracellularly

    • Prevents dehydration.

    • Increases resistance to antibiotics and disinfectants.

    • Slows nutrient uptake, thus inhibiting rapid growth.

    • Can evade destruction after phagocytosis due to unique cell wall properties.

  • Cultivation:

    • Growth rate is slow; it can take weeks to form visible colonies in laboratory settings.

    • Not susceptible to Gram staining due to cell wall structure.

Major Species Causing Human Diseases

  1. Mycobacterium tuberculosis

    • Responsible for tuberculosis (TB).

  2. Mycobacterium leprae

    • Causes leprosy.

  3. Mycobacterium avium-intracellulare

    • Affects immunocompromised individuals, particularly those with AIDS.

Mycobacterium tuberculosis Details

  • Historical Context:

    • A longstanding human pathogen evident in ancient remains (e.g., Egyptian and Native American mummies).

  • Disease Mechanism:

    • When the immune system is weakened, bacteria can escape containment, leading to active disease characterized by granuloma formation in the lungs.

    • Common in conditions of poverty, stress, malnutrition, and existing illnesses.

  • Public Health Concern:

    • Previously known as the "White Plague," tuberculosis can remain latent in 90-95% of infected individuals without causing illness for years.

  • Symptoms and Treatment:

    • Historical treatments included rest in sanitariums and exposure to fresh air to support the immune system against TB.

Conclusion

  • Understanding the characteristics and challenges posed by acid-fast bacteria such as Mycobacterium species is crucial for recognizing their roles in human diseases, particularly TB and leprosy.

  • Diagnosis of TB

    • PPD, Mantoux or Tuberculin Skin Test

      • A purified protein derivative of TB is injected under the skin.

      • If the person is infected, sensitized T cells cause a delayed hypersensitivity reaction.

      • Resulting area becomes red, hard, and swollen.

    • Limitations of PPD Test

      • Does not indicate whether TB is active or latent.

      • Requires an X-ray of the lungs to look for walled-off tubercles or active cavities.

    • Determination of Disease State

      • If tubercles are seen, next step is to determine if the disease is active or latent.

      • Check for Acid-Fast Bacilli (AFB) in sputum: presence of AFB indicates active TB.

    Laboratory Diagnosis

    • Culture Growth

      • Example shown: Mycobacterium tuberculosis on Lowenstein Jensen Agar.

      • Colonies resemble cauliflower.

    • AFB Tests

      • Special acid-fast staining techniques used to visualize Mycobacterium species in clinical samples (lungs, lymph nodes, etc.).

    Antibiotic Resistance and Treatment

    • Multidrug-Resistant TB (MDR TB)

      • Defined as TB resistant to the two most effective first-line drugs: isoniazid and rifampin.

    • Extensively Drug-Resistant TB (XDR TB)

      • MDR TB that is also resistant to effective second-line drugs commonly used for treatment.

    • Totally Drug Resistant TB

      • An extreme form of resistance with significant treatment challenges.

    • Directly Observed Therapy (DOT)

      • Involves a health worker visiting daily to ensure medication adherence, a strategy used to combat drug resistance.

    Vaccination Against TB

    • BCG Vaccine

      • Bacillus of Calmette & Guerin, a vaccine derived from M. bovis, a cattle pathogen.

      • Promotes immunity to TB and has been in use since the 1920s.

      • Commonly administered outside the U.S., but rarely used in the U.S. due to complications in testing (positive PPD from vaccination interferes with screening).

    Global TB Statistics

    • Incidence of TB

      • Annually, around 10 million people develop TB, with 1.5 million fatalities, making it the top infectious killer worldwide.

    • Vulnerability

      • Especially harmful to individuals with HIV.

      • Contributes significantly to antimicrobial resistance.

    • Geographical Distribution

      • Predominantly affects low- and middle-income countries, but TB exists globally.

      • Major burden concentrated in eight countries: Bangladesh, China, India, Indonesia, Nigeria, Pakistan, Philippines, and South Africa.

      • Gram Positive Rods

      1. Bacillus anthracis
      • Causes Anthrax:

        • Common disease in wild & domestic animals (outside the US).

        • Endospores can survive for years in the soil, waiting for grazing animals to ingest them.

        • Highest risk for humans are those in close proximity to infected animals.

        • Most common cause of anthrax in the US arises from imported hides or souvenirs made with animal skin.

      • Gram Stain Identification:

        • Presence of spores located in the center of the rods.

      2. Human Disease: Anthrax
      • Pulmonary Anthrax:

        • Most severe type of anthrax, occurs when endospores are inhaled.

        • Also known as "Woolsorters Disease" due to aerosolized spores from infected animal hides.

        • Initial symptoms similar to flu, progressive impairment of breathing.

        • Untreated case fatality rate: 90%.

      • Cutaneous Anthrax:

        • Occurs when broken skin contacts endospores, resulting in lesions.

        • Lesions can deepen, potentially leading to sepsis.

        • Most common form (95% of all cases).

        • Untreated case fatality rate: 20%.

      • Intestinal Anthrax:

        • Occurs if contaminated meat is ingested (rarest form).

        • Fatality rate ranges from 20-60% if untreated.

        • Mostly seen in predators consuming prey skin.

      3. Treatment/Transmission of Anthrax
      • Treatment Options:

        • Doxycycline, Ciprofloxacin, and Penicillin are effective.

        • Tetracycline and Erythromycin also have effectiveness.

      • Transmission:

        • Anthrax is not transmissible from person to person.

        • Could be used as a biological warfare agent; endospores need to be aerosolized for maximum effect.

      4. Bacillus cereus
      • Food Intoxication:

        • Produces two enterotoxins leading to two types of foodborne illness:

          • Heat-stable toxin: Rapid onset (1-6 hours), causing vomiting.

          • Heat-labile toxin: Onset (8-12 hours), causing diarrhea.

        • Associated with food like cooked rice or fried rice, which are held at room temperature too long before serving.

      • Mechanism:

        • Endospores survive boiling and germinate in favorable conditions, leading to multiplication at room temperature and toxin production.

        • Toxin remains viable, even if food is reheated.

      Notes on Bacillus cereus
      • No antibiotics needed for treatment of Bacillus cereus infections.

      • Spores do not cause the swelling of the cell.

  • Clostridium botulinum

    • Causes: Botulism, a severe form of food poisoning; can also contaminate wounds.

    • Exotoxin: Produces a potent neurotoxin, regarded as the most powerful natural toxin known; as little as 1µg can be fatal for a normal adult.

    • Symptoms:

      • Initial: Blurred vision and double vision.

      • Progression: Muscle weakness, difficulty swallowing, leading to potential respiratory or cardiac failure (flaccid paralysis).

    Botulism Types

    Adult Botulism
    • Pathway: Occurs primarily due to ingestion of canned food containing spores of C. botulinum that survive the sterilization process.

    • Growth: Spores germinate in the anaerobic environment of can, producing a dangerous toxin.

    Infant Botulism
    • Risk: Infants are more susceptible due to their immature intestinal flora compared to adults, which protects against the proliferation of C. botulinum.

    • "Floppy Baby Syndrome": Occurs when infants ingest honey or spores leading to botulism.

    • Precaution: Honey should never be given to babies since it may contain spores.

    Botox

    • Definition: A diluted form of botulinum toxin.

    • Medical Uses:

      • Effective for treating migraines, muscle spasms, excessive sweating, misaligned eyes, and some motion disorders like tip-toe walking.

      • Also popular in cosmetic treatments to reduce wrinkles.

    Clostridium perfringens

    • Role: Primary cause of gangrene, leading to tissue death from lack of blood supply.

    • Process: Gangrene initiates in anaerobic conditions of wounds where tissue necrosis assists bacterial growth, leading to rapid infection and gas production.

    Complications
    • Can cause severe uterine infections post-abortion due to its presence in the genital tract, which can lead to life-threatening sepsis.

    • If it originates from normal intestinal flora, perforation of the bowel may result in gangrene in the peritoneum.

    • Treatments: Penicillin or hyperbaric oxygen therapy; advanced cases may necessitate limb amputation due to necrosis.

    C. perfringens Food Poisoning
    • Prevalence: One of the most common causes of foodborne diarrhea in the U.S.

    • Mechanism: Bacteria thrive in the intestines and release enterotoxins, resulting in pain and diarrhea 8 to 12 hours post-consumption.

    • Association: Often linked to improperly refrigerated meat.

    Clostridioides difficile (formerly C. difficile)

    • Occurrence: Chief cause of hospital-acquired diarrhea and pseudomembranous colitis.

    • Characteristics:

      • Notable foul odor reminiscent of horse stables.

      • Usually part of the gut flora but can overgrow when antibiotics eliminate other microbes, particularly after using antibiotics like ampicillin or clindamycin.

    • Fatality Rate: Between 27-44% if untreated, affecting roughly 3.5 million North Americans annually.

    • Treatment: Standard options include metronidazole (Flagyl) or vancomycin (Vancocin, costs $1128).

    • Visual: Pseudomembranous colitis shows significant intestinal abnormalities compared to the normal colon lining.

    • Gram Positive Rods Overview

      • Definition: Bacteria that retain the crystal violet stain used in the Gram staining procedure.

      Endospore Forming Gram Positive Rods (GPR)
      • Characteristics:

        • Can survive extreme conditions such as:

        • Lack of food

        • Temperature extremes

        • Lack of moisture

        • Metabolically inactive when in spore form, making them resistant to destruction.

      • Examples:

        • Clostridium spp. (anaerobic)

        • Bacillus spp. (aerobic)

      Non-Endospore Forming Gram Positive Rods
      • Characteristics:

        • Lack the ability to form endospores.

      • Examples:

        • Corynebacterium

        • Listeria

        • Gardnerella (variable Gram staining)

        • Lactobacillus

        • Propionibacterium acnes

        • Actinomycetes (weakly acid-fast)

        • Mycobacterium (acid-fast)

      Clostridium Species
      • General Characteristics:

        • Anaerobic GPR that produce endospores and are associated with destructive toxins.

      Specific Clostridium Species:

      1. Clostridium tetani

        • Morphology: Round, terminal endospores resembling tennis rackets or lollipops.

        • Disease: Tetanus (caused by neurotoxin tetanospasmin)

        • Mechanism: Blocks muscle relaxation, leading to rigid contraction ("lockjaw") and muscle spasms.

      2. Clostridium botulinum

        • Morphology: Oval, sub-terminal endospores.

      3. Clostridium perfringens

        • Morphology: Oval, sub-terminal endospores, usually causing double zone hemolysis.

      4. Clostridioides difficile

        • Morphology: Oval, sub-terminal endospores, has a characteristic odor similar to horse manure.

      Tetanus (Clostridium tetani)
      • Pathology:

        • First Symptom: Stiffness of the jaw (lockjaw), progressing to severe muscle spasms.

        • Affected individuals may experience:

        • Rigid muscles in back and legs

        • Potentially fatal muscle spasms leading to respiratory failure.

      • Mechanism of Tetanus Toxin:

        • Toxin binds to inhibitory interneurons, reducing the release of neurotransmitters (glycine and GABA).

        • Inhibition leads to constant muscle contraction.

      Transmission and Infection Risks
      • Common Sources of Infection:

        • Soil, particularly around animal feces.

        • Entry routes:

        • Deep tissue injections (e.g., through puncture wounds)

        • IV drug use with non-sterile equipment.

        • At-Risk Populations:

        • 70% of cases in the U.S. are in individuals over age 50 whose vaccinations are not up to date.

      Tetanus Prevention and Treatment
      • Vaccination:

        • DTP Vaccine: Inactivated toxin that stimulates antibody production for immunity against tetanus.

        • Booster Shots: Required every 10 years (Tdap) to maintain immunity; many in the U.S. are unvaccinated.

      • Post-exposure Treatment:

        • Unvaccinated individuals may receive tetanus immune globulin (TIG) for immediate protection.

        • Antibiotics (e.g., metronidazole) can eliminate the bacteria.

      • Note: Awareness of symptoms is crucial for diagnosis, as cultures are only positive in 30% of cases.