Anaerobic Bacteria Collection

SPECIMEN COLLECTION GUIDELINES

Specimen Types

Site	Acceptable Specimens	Unacceptable Specimens
Head and Neck	Abscess (aspirate) Tissue (biopsy/surgically obtained) Fine needle aspirate	Throat/Nose/ Nasopharyngeal swabs Gingival/Mouth swabs/tongue Swabs from superficial areas
Eye	Aqueous/vitreous fluid Cornea/Corneal Rim/Donor Conjunctiva Slcera Eye tissue	Swabs from skin (e.g. eyelid) Eye drainage
Pulmonary	Lung aspirate/fine needle aspirate Tissue (biopsy/surgically obtained) Percutaneous lung puncture material Transtracheal aspirate (percutaneous) Bronchoscopic specimen (protected brush/double lumen) Deep bronchial secretion Thoracotomy specimen	Sputum (expectorated/ induced) Endotracheal/ Tracheostomy aspirate Bronchial wash BAL Bronchoscopic specimens not specially collected Nasotracheal tube suctioning
Abdomen	Peritoneal fluid Abscess aspirate (surgical) Tissue (biopsy/surgically obtained) Appendiceal tissue or abscess Gallbladder Liver Bile Pancreas Biloma Ascites	Aerobic swabs Drainage Gastric/Bowel contents Ileostomy/ Colostomy effluents Gastric washing (unless newborn) Perineal/Rectal swabs Feces Perirectal and rectal abscess NOT obtained by surgical procedure Decubitus ulcers NOT obtained by surgical procedure
Genital and Reproductive	Culdoscopy specimen Endometrial aspirate Abscess aspirate (needle/syringe) Biopsy surgically obtained IUD for Actinomyces species Bartholin's’gland Fallopian tube fluid/tissue Placenta tissue (via Cesarean)	Vaginal/Cervical swabs/secretions Prostatic secretion Lochia secretions Seminal fluid
Joint	Aspirate obtained by needle/syringe Biopsy surgically obtained	Superficial material on swabs
Sterile	Bone/Bone marrow (biopsy/aspirate) Brain abscess Pleural fluid Pericardial fluid CSF, shunt Tissue (biopsy/surgically obtained) Aspirate by needle/syringe Deep aspirate (decontaminated skin)	Superficial material (e.g. skin) Aerobic swabs CSF, lumbar puncture
Urinary Tract	Suprapubic aspirate Cystoscopy Ureter aspirate Kidney aspirate	Void urine Catheterized urine Nephrostomy Bricker Ileal bladder
Miscellaneous	Other fluid (large volume) Rectal abscess Perirectal abscess Pilonidal cyst Miscellaneous skin soft tissue	Autopsy specimens Decubitus ulcer material Drain/Drain site Exudate

Containers

Ideally, pus, fluid and/or tissue (not swabs) should be collected to optimize the recovery of the anaerobic bacteria present. An anaerobic transport tube ^(a) or jar ^(b) should be used – pus or fluid can be directly injected into this container using a needle and syringe. Never send a needle and syringe to the laboratory – this is unsafe and unacceptable practice. Needles should be removed or capped with a point-lok device. Tissues can be placed into the anaerobic tube or jar (preferred) by momentarily unscrewing the cap, holding the tube or jar upright at all times and inserting the sample into the agar (approximately 5 mm from the bottom of the jar). The cap should be immediately replaced and screwed down tight for transport of the tube. 
 
Anaerobic transport tubes (^a = Anaerobic Transport Medium Surgery Pack, Cat # AS-914) and jars (^b = BBL Port-A-Cul Sterile Pack Transport Jar, Cat # 221602) are stocked in the hospital on all nursing stations, the Emergency Department and in the Operating Rooms. In the community, physicians can order anaerobic collection kits using the standard forms for other collection materials and supplies. Use Lawson numbers stated above. Always ensure that the correct collection kits are available before performing a procedure that may require specimens to be sent for anaerobic culture.

Turn Around Time

Anaerobes grow slowly, so that the isolation and identification of an organism from an anaerobic culture may not be finalized for up to 7 days. If susceptibility testing is requested, results may take 7-14 days since this testing is performed by an outside reference laboratory.

Order and Specimen Handling

Label the tube and order the test. If using WellSpan's Epic, follow protocol to order the test and generate a computerized specimen label. If not using Epic, label the tube and complete a Microbiology request form. The requisition for anaerobic culture should be fully completed and placed in the outer pouch of a Biohazard transport bag. Place the anaerobic tube or jar in the inner compartment of the bag. Make sure the specimen is immediately transported to the lab. 
  
A sufficient volume or amount of patient specimen must be submitted in an oxygen-free transport system to the microbiology laboratory to permit recovery of anaerobic pathogens. For tissues, the ideal amount is approximately 1 gram and for fluids, the ideal volume is 1 mL.  Swab specimens are unacceptable for recovery of anaerobes unless they are embedded in a special anaerobic transport medium. For optimal recovery of potentially pathogenic bacteria, the laboratory recommends the following collection methods:

PROCEDURES FOR COLLECTING/TRANSPORTING ANAEROBIC SPECIMENS

MOST ACCEPTABLE METHODS

Sterile Anaerobic Transport Medium Method (Anaerobe Systems #AS914). Use this method for tissue or bone specimens.

• Medium
  • Must be stored at room temperature.
  • Contains both thioglycollate and cysteine in an agar base to reduce the oxygen tension of the specimen.
• Equipment needed
  • Anaerobic Transport Medium tube
  • Gloves (sterile or non-sterile depending upon procedure)
  • Completed lab requisition (if not using Orders Mnagement)
• Procedure
  • Check patient identiband.
  • Explain procedure to patient; screen as necessary.
  • Wash hands.
  • Apply gloves.
  • If applicable, remove dressing and place in plastic bag (see procedure for dressings).
  • Obtain the specimen (cut tissue or bone) that is small enough to fit within the Anaerobic Transport tube. Note: A small area representing the focal point of infection is sufficient for culture.
  • Holding the transport tube upright, open the capped tube carefully and quickly.
  • Quickly place the tissue or bone on top of the agar.
  • Quickly recap the tube.
  • Remove gloves, wash hands.
  • Redress wound if applicable.
  • Label the tube and order the test:
  • Transport the specimen to the laboratory.
    • If using the pneumatic tube system, follow the packaging instructions for transporting items via the system.
    • If transporting via courier:
      • Place the Anaerobic Transport Medium tube in a specimen transport bag.
      • Place the transport bag in a transport cooler.

Note: An alternative transport system for tissue or bone specimens is the larger BBL™Port-A-Cul™ Sterile Pack Transport Jar:

• Store Port-A-Cul at room temperature prior to use.
• Do not open the envelope until ready to use.
• Do not use if medium in unopened jar is oxidized (pink to lavender color) to a depth greater than 3 mm.
• Open package by peeling apart.
• Obtain specimen.
• Using aseptic technique, remove jar cap and immediately insert specimen approximately 5 mm from the bottom of the jar.
• Replace screw cap and tighten.
• Transport to the laboratory.

Syringe/Needle Method Use for abscesses, sinus tracts, draining wounds.

• Equipment needed:  You may use EITHER one.
  • Sterile 3 mL or 10 mL syringe.
  • 22 gauge needle attached.
  • Stire cap to close the syringe tip
• Procedure
  • Confirm patient's identity with at least two identifiers.
  • Explain procedure to patient; screen as necessary.
  • Wash hands.
  • Apply gloves.
  • If applicable, remove dressing and place in plastic bag (see procedure for dressings).
  • Decontaminate skin, but do not introduce disinfectant into the collected sample.
  • Aspirate the specimen fluid into the syringe avoiding the introduction of room air.
  • Expel any air, cap and remove needle, and close the syringe with a sterile plastic cap.
  • Remove gloves, wash hands.
  • Redress wound if applicable.
  • Label the syringe and order the test.
  • Transport the specimen to the laboratory
    • If using the pneumatic tube system, follow the packaging instructions for transporting items via the system.
    • If transporting via courier:
      • Place the labeled, capped syringe in a specimen transport bag.
      • Place the transport bag in a transport cooler.
      • Give the completed Microbiology request form (if using one) to the courier.

LEAST ACCEPTABLE METHOD

Swab Method (Note: Swabs expose anaerobic bacteria to oxygen and must ONLY be used for recovery of anaerobes if the two methods described above cannot be used.)

• Equipment
• Aerobic culture swab and Anaerobic Transport Medium Tube

OR 

• E-swab

• Procedure
  • Confirm patient's identity with at least two identifiers.
  • Explain procedure to patient; screen as necessary.
  • Wash hands.
  • Apply gloves.
  • If applicable, remove dressing and place in plastic bag (see procedure for dressings).
  • Open the aerobic or E-swab culture swab and carefully remove the swab (discard the aerobic culture tube).
  • Swab the area.
  • Holding the anaerobic transport medium tube upright, carefully remove the cap.
  • Quickly embed the swab just below the surface of the agar in the tube. Do not push the swab to the bottom of the tube. For E-swab, quickly place swab in tube and replace cap.  
  • Quickly break the shaft of the swab off at the lip level of the transport tube using a sharp, snapping motion.
  • Quickly replace the cap.
  • Remove gloves, wash hands.
  • Reapply dressing, if applicable.
  • Label the Anaerobic transport tube and order the test.
• Transport the specimen to the laboratory
• • If using the pneumatic tube system, follow the packaging instructions for transporting items via the system.
  • If transporting via courier:
    • Place the labeled tube in a specimen transport bag.
    • Place the transport bag in a transport cooler.
    • Give the completed Microbiology request form (if using one) to the courier.

Note:  Additional swabs under aerobic conditions must be sent for stains and recovery of aerobic bacteria.