Microbiology
- Bacteria
- prokaryotes (lack a nucleus and chromosomes)
- classified morphologically as rods, cocci, or spirals
- also classified according to the Gram-staining characteristics of their cell walls
- external to the cell membrane, bacteria have a cell wall that is much thicker in gram-positive than
in gram-negative bacteria
- many antibiotics work by inhibiting cell wall synthesis
- gram-negative bacteria, in addition to their cell wall, have an outer membrane containing
lipopolysaccharide (endotoxin)
- in some bacteria, surrounding the cell wall is a thick gel-like structure called a capsule
- capsules appear to protect the bacteria from phagocytosis, thereby acting as a virulence factor
- Fungi
- eukaryotic cells (contain a nucleus with a nuclear membrane)
- contain a cell wall that is external to the plasma membrane
- also have a capsule that inhibits phagocytosis
- some fungi are primary pathogens (Histoplasma, Blastomyces, Coccidiodes)
- other fungi cause opportunistic infections (Candida, Aspergillus, Cryptococcus)
- Candida albicans is the most frequent clinical isolate
- Protozoa
- eukaryotes
- human pathogens include Pneumocystis carinii and Trichomonas vaginalis
- Viruses
- most primitive form of infectious pathogen
- obligate intracellular parasites
- must invade cells and use their energy-generating machinery in order to replicate
- Hepatitis B, C and HIV infections may be transmitted from patients to health care workers
Determinants of Infection
- Microbial Pathogenicity
- ability of a microbe to cause infection is a balance between host defenses and microbial pathogenicity
- some bacteria possess specific characteristics that make them virulent
- Virulence Factors
- Exotoxins
- some bacteria can secrete highly potent toxins
- Clostridium tetani produces a potent neurotoxin
- Clostridium botulinum produces a neuromuscular toxin
- S. aureus is responsible for toxic shock syndrome
- Group A Streptococci secrete potent hemolysins
- Endotoxins
- lipopolysaccharide-protein complexes that are normal constituents of the cell wall
of gram-negative bacteria
- Lipid A appears to be the toxic portion of LPS
- activates both the intrinsic and extrinsic coagulation cascades
- activates the alternative pathway of the complement cascade
- activates macrophages and stimulates the production of numerous cytokines
(IL-1, IL-6, TNF)
- Additional Virulence Factors
- many bacteria and fungi have thick capsules, making them resistant to phagocytosis
- S. epidermidis secretes a ‘slime’ that facilitates its binding to foreign
surfaces and makes it less vulnerable to phagocytic cells
- some bacteria produce various enzymes that thwart the bactericidal mechanisms of
phagocytic cells
- collagenase produced by streptococci and staphylococci facilitates
dissection of infection along fascial planes
- Antibiotic Resistance
- each group of antibiotics has a different mechanism of action that exploits a
metabolic or structural vulnerability of the sensitive organism
- many antibiotics interfere with cell wall synthesis; others inhibit protein
synthesis by targeting bacterial ribosomes
- antibiotic resistance may occur through several different mechanisms: plasmids
(most common) and spontaneous mutations (rare)
- Host Defenses
- Local Host Defenses
- Skin
- principle defenses of the skin are dryness and desquamation
- bacteria need moisture to grow and normal skin has a low moisture content
- keratin constantly sloughs, and the flora is sloughed as well
- sebaceous glands also secrete a lipid inhibitory to most pathogens
- colonization is sparse on normal skin except in intertriginous areas where moisture
accumulates
- Respiratory Tract
- most inhaled particles, including microbes, never reach the alveoli
- large particles are filtered out in the nares
- mucous layers in the trachea and bronchi contain a number of defenses (IgA,
lysozyme, complement)
- cilia move the mucous layer towards the mouth, where it is expelled by coughing
- small particles that make it to the lower airway are phagocytized by pulmonary
alveolar macrophages
- Alimentary Tract
- saliva possesses lysozyme and IgA
- ingested organisms are killed in the highly acidic stomach
- peristalsis in the small intestine makes it difficult for bacteria to adhere,
although they do proliferate
- drugs, operations, or disease states that diminish gastric acidity allow bacteria to
proliferate in the stomach
- prolonged ileus will also allow bacteria to overgrow in the small intestine
- Large Intestine
- low-oxygen, low motility environment of the colon allows exponential growth
of bacteria
- anaerobes outnumber aerobes by 100:1
- only a few of these species are ever isolated in intra-abdominal infections
- normal colonic flora suppresses growth of pathogenic species
- prolonged use of broad-spectrum antibiotics will greatly reduce the normal
colonic flora
- Urinary Tract
- protected by periodic flushing of urine
- urine is somewhat bacteriostatic by virtue of its high urea content, high osmolarity,
acidic pH
- long urethra in males limits microbial access
- short female urethra facilitates microbial access
- urinary catheters introduce bacteria into the bladder
- Systemic Host Defences
- Neutrophils (PMNs)
- once microbes have breached the skin or mucosal barrier, the most important host
defense against infection is the phagocytic system (innate response)
- PMNs respond long before the production of specific antibody and cell-mediated
responses
- phagocytes must recognize that an organism is ‘bad’
- this is mediated by opsonins
- opsonins bind to the microbial surface and then to specific receptors on the
phagocytic cell membrane
- Macrophages
- also act as phagocytic killers
- found throughout the body’s tissues
- secrete a vast array of cytokines
- function as antigen-presenting cells for lymphocytes
- Complement System
- activated by direct contact with microbes and via IgM > IgG binding to microbes
- also activated when the coagulation cascade is activated
- membrane attack complex destroys pathogens
- some complement protein fragments are biologically active (C3a, C5a)
- Local Environmental Factors
- Bacterial Load
- a critical threshold of bacteria is necessary to cause clinical infection
- in general, 100,000 bacteria per gram of tissue must be present to cause a clinical
infection
- more virulent bacteria may require much fewer numbers to cause severe or
life-threatening infections (clostridial gangrene or tetanus)
- the presence of adjuvant factors may also lower the number of bacteria required to
cause infection
- Adjuvant Factors
- amplify bacterial virulence or repress host responsiveness
- hemoglobin (clot or hematoma) may be the most potent adjuvant factor
- foreign bodies (suture, prosthetic materials)
- necrotic tissue
Common Surgical Pathogens
- Staphylococci
- S. aureus
- possesses virulence factors which permit it to invade normal tissues
- skin organism found in the anterior nares and areas of moist skin in up to 40% of healthy
people
- suture material decreases by 500 times the number of bacteria required to produce an infection
- most common isolate from wound infections after clean procedures
- most hospital strains are methicillin-resistant (MRSA)
- MRSA
- vancomycin is the standard first-line therapy
- linezolid (Zyvox) is an oral agent useful for outpatient treatment
- S. epidermidis
- normal resident of the skin
- causes infection in the presence of foreign bodies such as intravenous catheters, heart
valves, prosthetic joints
- some strains produce a glycocalyx (‘slime’), which allow bacteria to resist phagocytosis and
adhere to prosthetic materials
- Streptococci
- classified according to cell surface antigens and their ability to cause hemolysis on blood agar
- Group A Streptococci
- comprised of a single species: Strep pyogenes
- β-hemolytic
- responsible for necrotizing soft tissue infections and erysipelas (streptococcal cellulitis
and lymphangitis)
- may secrete a number of exotoxins
- hyaluronidase and streptokinase promote the spread of infection along tissue planes
- Group B Streptococci
- harbored in the female genital tract
- responsible for neonatal meningitis and bacteremia
- Strep. pneumoniae
- common resident of the nasopharynx
- possesses a capsule with antiphagocytic properties
- responsible for overwhelming post splenectomy sepsis
- antipneumococcal vaccine is 65% to 70% effective in preventing infection
- Enterococcus
- part of the normal flora of the GI tract and vagina
- often found in polymicrobial infections of the peritoneum and pelvis
- may also cause UTIs and endocarditis
- now resistant to many antibiotics, including vancomycin (VRE)
- VRE may be treated with linezolid, daptomycin, or tigecycline
- Neisseria
- gram-negative cocci
- N. gonorrhoeae is harbored in the urethra, cervix, pharynx, conjunctiva
- transmission is usually by sexual contact
- contiguous spread from the endocervix can result in PID
- spread of gonococci into the upper abdomen can result in perihepatitis (Fitzhugh-Curtis syndrome)
- Gram-Negative Bacilli
- Enterobacteriaceae
- facultative aerobes
- commensal in the GI tract and vagina
- contain endotoxin in their cell walls
- E. coli is the most common GNR in the GI tract and is the most common organism isolated from
biliary, urinary, and intra-abdominal infections
- Klebsiella is an encapsulated organism commonly associated with hospital-acquired pneumonias
- multidrug resistant organisms are becoming more common
- Pseudomonads
- obligate aerobes
- able to survive in water almost without other nutrients
- normal host defenses are highly effective against colonization
- colonization occurs in debilitated patients whose own flora has been depleted by antibiotics
- pseudomonads flourish in ICUs when they contaminate wet equipment (respirators) and are
spread by personnel from patient to patient
- Pseudomonas aeruginosa is the leading cause of death from nosocomial infections in the ICU
- Anaerobic Bacteria
- require a low-oxygen environment for growth
- found predominantly in the mouth, GI tract, and vagina
- in most infections, aerobic bacteria are also present
- anaerobes outnumber aerobes by 10 to 1 in the mouth and by 100 to 1 in the colon
- predisposing conditions for anaerobic infection include poor vascular supply, tissue necrosis,
foreign bodies
- Bacteroides
- low virulence organisms
- have an endotoxin, but it is of poor biologic activity
- their slow growth in culture (up to one week) may delay their diagnosis and make empiric
treatment necessary
- Clostridia
- gram-positive spore-forming rods
- most virulent anaerobic species
- all are soil organisms and are found in the gut as normal flora
- C. tetani
- tetanus occurs in nonimmunized or partially immunized individuals who have
contaminated wounds
- produces an exotoxin that causes muscle spasms
- treatment involves penicillin to eradicate the source of the toxin and tetanus
immune globulin to neutralize circulating and unbound toxin
- C. perfringens
- causes necrotizing soft tissue infections
- secretes many exotoxins
- C. difficile
- produces an exotoxin that causes pseudomembranous colitis
- occurs in patients who have received broad-spectrum antibiotics
- oral vancomycin or metronidazole are effective therapies
- Fungi
- fungal surgical infections are caused primarily by the opportunistic fungi
- Aspergillus
- colonizes the upper airway in healthy individuals
- in the compromised host, may infect the ear, sinuses, orbit, lung, and brain
- Mucormycosis
- caused by fungi of the mucor and rhizopus genera
- occurs in the context of uncontrolled diabetes, immunosuppression, thermal burns
- begins in the nose and may spread contiguously to the orbits, palate, eye, and brain
- treatment consists of aggressive surgical debridement and amphotericin B
- Candida
- normal inhabitant of the mouth, upper airways, GI tract
- overgrowth occurs when competing bacteria are eliminated by antibiotic therapy
- risk factors for invasion include malignancy, malnutrition requiring hyperalimentation,
diabetes, systemic steroids, immunosuppression
- Viruses
- Hepatitis
- Hepatitis B
- DNA virus
- infection occurs through needle sticks, exposure to blood, sexual contact
- needle sticks cause a 30% conversion rate in unvaccinated individuals
- ~5% of those infected become chronic carriers
- with blood bank screening, hepatitis B now rarely accounts for post transfusion
hepatitis
- hepatitis B vaccine is available and is 95% protective
- 250,000 cases/year in the 1980s; 3350 cases in 2010
- hepatitis B immune globulin is available for post exposure prophylaxis in individuals
not vaccinated against hepatitis B
- Hepatitis C
- RNA virus
- 2% conversion rate from a needle stick
- no vaccine available
- most patients are asymptomatic
- progression to chronic persistent and chronic active hepatitis occurs in 75% of
patients and may progress to cirrhosis
- first or second most common indication for liver transplantation
- rarely transmitted through blood transfusions now because of sensitive blood bank
testing
- interferon alpha and ribavirin are used to successfully treat chronic hepatitis C
- Cytomegalovirus
- member of the herpes virus family
- often complicates organ transplantation
- commonly transmitted during blood transfusions
- transfusion of CMV-seropositive blood to an immunocompetent patient will result in only a
mild infection, if any
- diagnosis is made by demonstrating intranuclear inclusions (owl’s eyes) in tissue sections
- immunocompromised patients may be treated with ganciclovir
- Human Immunodeficiency Virus (HIV)
- Microbiology
- RNA retrovirus with an affinity for the CD4 receptor on T lymphocytes
- after binding to the CD4 receptor, the virus is internalized and uncoated
- reverse transcriptase synthesizes DNA complementary to the viral RNA
- this DNA is then incorporated into the host cell’s genome, leading to a lifelong
infection
- Pathophysiology
- infection with HIV leads to T helper cell deficiency (CD4), resulting in profound
depression in cell-mediated immunity
- once the CD4 lymphocyte count drops below 200/μL, the chance of developing an
opportunistic infection is high
- Serologic Events
- 6 to 12 weeks after infection, antibody to HIV develops (may take up to 6 months)
- serologic testing examines antibodies to HIV
- since it may take up to 6 months to develop antibodies to HIV, during this time a
patient may have circulating virus and yet test negative for HIV
- Surgery in HIV-infected Patients
- patients do not require any special preoperative preparation
- prophylactic antibiotics are given for the same indications as for non-HIV patients
- these patients do not have difficulty with wound healing and do not have a higher
rate of infectious complications
- HIV Post Exposure Management
- risk of developing seropositivity after a needle stick is about 0.3%
- blood from the source patient should be drawn and tested for HBV, HCV, and HIV
- following the initial test, seronegative workers should be retested at 6 weeks,
12 weeks, and 6 months
- if the source patient is HIV positive, post exposure prophylaxis is associated with
a lower risk of HIV transmission
- if the HIV status of the source patient is unknown, the use of post exposure
prophylaxis should be decided on a case-by-case basis (high risk vs low risk)
Specific Infections
- Wound Infections (SSIs)
- Wound Classification
- Clean
- elective, primarily closed, undrained
- nontraumatic, uninfected
- respiratory, alimentary, GU, oropharyngeal tracts not entered
- infection rate of 1.3% - 2.9%
- hernia, breast biopsy
- Clean Contaminated
- alimentary, respiratory, GU tracts entered under controlled conditions
- mechanical drainage
- minor break in technique
- elective GI tract surgery (not colon), elective cholecystectomy
- infection rate of 2.4% - 7.7%
- Clean Contaminated – Colorectal
- infection rate of 4% - 14% in adequately prepped patients
- Contaminated
- open, fresh traumatic wound
- gross spillage from the GI tract
- biliary surgery in presence of infected bile
- incision through inflamed, but nonpurulent tissue
- penetrating abdominal trauma
- infection rate of 6.4% - 15.2%
- Dirty wounds
- perforated viscus
- pus encountered
- perforated diverticulitis, necrotizing STI
- infection rate of 7.1% - 40%
- Prophylaxis
- SSIs are associated with increased morbidity/mortality, patient dissatisfaction
- also associated with substantial costs to the health care system
- many payers are refusing to pay hospitals for SSIs
- all hospitals that participate in Medicare must report SSIs within 30 days of the procedure
or 90 days if prosthetic material is implanted
- good glucose control may minimize SSIs
- intraoperative hypoxia and hypothermia should be prevented
- Operating Room Environment
- air filtration systems can reduce the number of dust particles to which microbes can
adsorb
- operating rooms should have a positive pressure relative to air in the corridor so
that unfiltered air does not enter the OR
- laminar flow rooms are used when prosthetic joints are implanted
- Patient Preparation
- preop shower with an antiseptic soap can reduce the resident skin bacteria
- however, the evidence is unclear whether this reduces wound infections
- elective operations should be delayed until remote site infections have been cleared
up
- shaving should be done in the OR with clippers, not the night before
- skin should be prepped with povidone-iodine or chlorhexidine
- bowel prep prior to elective colon surgery
- Surgical Technique
- meticulous hemostasis
- remove all devitalized tissues
- Prophylactic Antibiotics
- indicated in most clean-contaminated cases
- also indicated in clean operations in which an infection could lead to a disastrous
outcome, such as prosthetic joint, vascular graft, or cardiac valve procedures
- choose an antibiotic effective against the pathogens most likely to be encountered
and has low toxicity
- should be given 30 to 60 minutes prior to the incision
- give a second dose if the procedure lasts longer than 4 hours
- should be discontinued within 24 hours of surgery
- cefazolin is active against most gram-positive organisms (not MRSA) and some
gram-negative organisms
- second-generation cephalosporins have broader coverage against gram-negative
organisms and also have GI anaerobic coverage, making them useful in colon surgery,
appendectomy
- for patients with a true penicillin allergy, vancomycin or clindamycin can be
substituted
- Management
- primarily consists of opening and packing the wound
- antibiotics are reserved for when significant cellulitis or fever is present
- wound VACs can be very helpful in managing large or complex wounds
- Central Line Infections
- Prevention
- hand washing
- full barrier precautions (cap, mask, sterile gloves, sterile gowns, and sterile drapes)
lower line infection rates dramatically
- skin preparation with chlorhexidine has lower infection rates when compared with
povidone-iodine solutions
- subclavian vein insertion site has the lowest infection rate; the femoral vein has the
highest
- antibiotic-impregnated catheters have lower infection rates in catheters left in place for
more than 5 days
- multilumen catheters have higher infection rates
- dedicated catheter care teams results in fewer catheter-related infections
- central lines should be removed when they are no longer clinically necessary
- Replacement of Central Lines
- routine replacement of lines that have no signs of infection is not recommended,
except in high-risk populations like burn or transplant patients
- replacing a line over a guidewire should rarely be done because of higher infection
rates
- Diagnosis
- a high index of suspicion is necessary
- purulence and inflammation around the insertion site is a specific but uncommon finding
- fever is a common finding, but it is not very sensitive for line infections
- definitive diagnosis requires peripheral blood cultures to grow the same organism as the
catheter tip
- if clinical suspicion is high, then the line should be removed and the tip sent for culture
- Microbiology of Central Line Infections
- S. aureus and S. epidermidis originate from the skin and cause most catheter-related
infections
- most candida infections result from hematogenous dissemination
- Treatment
- Antibiotic Selection
- vancomycin is the preferred empiric drug because most hospital staph species are
methicillin resistant
- daptomycin can be substituted if vancomycin is contraindicated
- pseudomonas coverage can be added in high risk patients
- fluconazole is the treatment choice for candida infections
- Catheter Removal
- the great majority of vascular catheter infections will require removal of the
catheter
- catheter infections with S. epidermidis can occasionally be treated with antibiotics
instilled into the catheter and long-term systemic antibiotics
References
- Schwartz, 10th ed. Pgs 135 -157
- O’Leary, 4th ed. Pgs 218 - 257
- Simmons and Steed, pgs 56 – 83
- Sabiston, 20th ed., Pgs 240 - 250
- UpToDate, Antimicrobial Prophylaxis for Prevention of Surgical Site Infection in Adults.
Anderson, Deverick. March, 2018. Pgs 1 – 48.
- UpToDate, Overview of Control Measures for Prevention of Surgical Site Infection in Adults.
Anderson, Deverick. October 2018, pgs 1 - 34
- Cameron, 11th ed., pgs 1259 - 1262