APPLIED MICROBIOLOGY

Septicaemia and Bacteraemia

• Septicaemia literally means 'sepsis of blood'. It is a condition in which there is presence of actively
multiplying bacteria in the blood stream, and formation of toxic products in the blood.
• Bacteraemia is a condition in which there is transient presence of bacteria in blood stream without
multiplication. Clinical signs of septicaemia are absent.
• Toxaemia refers to formation of toxic products in blood. When bacterial endotoxins circulate in the
blood, the condition is called endotoxaemia. Endotoxaemia can occur in absence of bacteria in the blood
stream, e.g. from Gram-negative bacteria present in the gut, endotoxin passes across the gut wall.

Factors predisposing to septicaemia are, as follows:

I. Impaired Host Defenses, for e.g.
• Deficiency of cell-mediated/ humoral immunity
• Decreased complement level etc.
• Diminished inflammatory response
• Diabetes mellitus
• Malignancy
• Prematurity and low birth weight
• Intravenous drug abuse
• Old age
• Immunosuppressive therapy
• Splenectomized patients
• Uraemia
• Hepatic failure
• Aplastic anaemia
• Myeloma

II. Pre-existing Infections

III. Instrumentation and Surgery Examples
IV. Other Factors Such as-difficult delivery, exposure to infected incubators, exposure to infected
ventilators, etc.

Following are the microorganisms causing septicaemia and bacteraemia:

Bacteria:
· Escherichia coli (22 %)
· Streptococcus pneumoniae (17%)
· Staphylococcus aureus (14%)
· Klebsiella sp. (7%)
· Proteus sp. (5%)
· Haemophilus influenzae (4%)
· Salmonella sp. (4%)
· Streptococcus pyogenes (4%)
· Pseudomonas aeruginosa (3 % )
· Group B Streptococci (2%)
· Str. viridons (2%)
· Staph. epidermidis
· Enterococci
· Serratia marcescens
· Enterobacter aerogenes
· Citrobacter sp.
· Acinetobacter sp.
· Bacteroides sp.
· Fusobacterium sp.
 · Anaerobic cocci-peptococci and peptostreptococci
· Neisseria meningitidis
· Legionella sp.
· Brucella sp.
· Bacillus sp.
· Rickettsia sp.
· Coxiella sp.
· Leptospira sp.
· Borrelia sp.
· Yersinia sp.
· Francisella sp.
· Streptococcus Group C and G

Clinical Features:

· Fever or hypothermia
· Rigors
· Tachycardia
· Tachypnoea
· Hypoxia
· Dyspnoea
· Cyanosis
· Hypotension
· Mental confusion
. Agitation in elderly patients
· Behavioural

Complications:

· Septic, endotoxic or bacteraemic shock

· Disseminated intravascular coagulation (DIC)
· Acute renal failure
. Shock may lead to multiple organ failure (e.g. heart, lungs, liver and kidneys)

Laboratory diagnosis of septicaemia/bacteraemia:

Blood Culture
• Collect 10 ml of blood with the help of sterile syringe and needle
• Dilute the sample (1: 1 0) to inactivate antimicrobial components present. 5 ml of blood is
collected in 50 ml of glucose broth and 5ml in 50 ml of bile broth.
• Brain-heart infusion broth, trypticase soya broth and even nutrient broth can be used
• For anaerobes-thioglycollate broth or Robertson's cooked meat medium can also be used
• Sodium polyanethol sulphonate (SPS) is added to broth media, which acts as an anticoagulant,
antiphagocytic, anticomplementary. It interferes with the activity of certain antimicrobial agents
such as aminoglycosides, enhances growth rate and increases the rate of isolation

After collection, blood culture bottles are incubated aerobically and blind subcultures are made after 24
hours, 48 hours, 72 hours, on 6th day and 10th day. The subcultures are made on:
• Blood agar
• MacConkey's agar
• Chocolate agar
• Anaerobic supplemented blood agar for anaerobes
• Sabouraud's dextrose agar for fungi
• Other selective media as indicated
After incubation, colonies obtained on solid media are studied further for identification by using
biochemical reactions and antibiotic susceptibility by Kirby-Bauer's disc diffusion method

Castaneda's Method
 This method is used for rapid detection of causative agents, to minimize the materials and
manipulations required for processing of specimen, to reduce the problem of contamination and to
reduce the risk of infection to laboratory workers
Castaneda's medium is a diphasic medium containing both liquid and solid media in the same bottle; for
example, it contains brain-heart infusion broth and agar in the same bottle. The blood specimen is
collected into liquid medium and bottle is incubated in upright position. For inoculation of agar, bottle is
tilted at intervals so that the broth flows over the agar slant. Again it is incubated in upright position.
The colonies formed on agar slant are studied and identified.

Automated Blood Culture System:

Various newer unconventional methods for detecting bacteria in blood are now available which detect
bacterial growth by various techniques such as the detection of carbon dioxide by radiometric or optical
methods. One such instrument commonly used is Badec System

Non-culture Methods
• Various nonculture methods are used to detect circulating antigens and other microbial products, as:
Latex agglutination test-used for detecting antigen of Group B Streptococci, H. influenzae type b, Str.
pneumoniae, N meningitidis, staphylococci and some yeasts
• Counter current immunoelectrophoresis (CIEP)-used in detection of pneumococci, Klebsiella,
H.influenzae and other microbial agents
• Limulus amoebocyte lysate assay test-used to detect circulating lipopolysaccharide (endotoxin) of Gram-
negative bacteria in blood
• Gas liquid chromatography-detects metabolic end products of microbial activity in serum

Septicaemia/bacteraemia can be treated in following ways:

• Immediate parenteral antibiotic therapy (IV route is the best) in adequate dosage necessary as soon as
specimen for culture is collected
• Surgical intervention-urgent surgical drainage of abscesses, wounds, etc.
• Intravenous fluids to prevent shock
• Blind drug therapy is given before the availability of antibiotic susceptibility testing report, based on the
- Clinical symptoms and signs
-Likely focus and cause of infection
-Knowledge of the underlying conditions
• The correct antibiotic therapy is started after the availability of antibiotic susceptibility testing report
• The antibacterial agents used for blind drug therapy include:
o Penicillin
o Cloxacillin for Gram-positive bacteria
o Fusidic acid
o Vancomycin
o Ampicillin
o Gentamicin
o Carbenicillin
o Tobramycin for Gram-negative bacteria
o Azlocillin
o Cefotaxime .
o Metronidazole - for anaerobes
Combination of drugs can be given in combined infections.