Gauze (woven or pressed)
3. Applicator sticks (wood)
4. Sterile cryovials or screw-cap vials (to hold 1 ml)
5. Box for vial storage in freezer
6. Filter paper, Whatman no. 1 and Whatman no. 42
7. Sterile filtration system
8. Membrane filters (pore size, 0.22 µm) (to be used with sterile filter)
9. Nuclepore membrane filter, 25-mm, 5-µm, and 3-µm porosity
10. Swinney filter adapter (attaches to syringe, holds filter)
11. Filter paper pad, 25 mm (used to support the membrane filter in the Swinney adapter)
12. Bacteriological loop
13. Sterile syringe needles, 20 and 27 gauge
14. Vaspar
15. Parafilm (American Can Co.) or equivalent
16. Slide boxes for positive-slide storage
17. Forceps and scissors
18. Stage micrometer with scale of 0.1- and 0.01-mm divisions
19. Disk micrometer divided into 50 units
20. Biohazard container with disinfectant for proper disposal of slides, tubes, and pipettes
21. Biohazard container for proper disposal of patient specimens
22. Microscope lens paper
ATCC Quality Control Organisms
1. An 18- to 24-h-old culture of Escherichia coli or Enterobacter aerogenes
2. ATCC 30010 (Acanthamoeba castellanii)
3. ATCC 30133 (Naegleria gruberi)
4. ATCC 30925 (Entamoeba histolytica HU-1:CDC)
5. ATCC 30015 (Entamoeba histolytica HK-9)
6. ATCC 30001 (Trichomonas vaginalis)
7. ATCC 30883 (Leishmania mexicana)
8. ATCC 30160 (Trypanosoma cruzi)
Note None, some, or all of these QC organisms will be required, depending on the culture procedures performed in the laboratory.
Safety: Personnel and Physical Facilities
1. Be careful! All material to be received by or discarded from the laboratory must be considered potentially pathogenic.
2. Smoking, eating, or drinking in the laboratory is not permitted.
3. Do not work with uncovered open cuts or broken skin. Cover them with a Band-Aid, finger cot, or other suitable means, such as rubber or plastic gloves.
4. Mouth pipetting of specimens is not permitted.
5. Do not create aerosols. Remember, infectious diseases, such as infectious hepatitis, may be transmitted by aerosols produced by centrifuges, stirrers, pipettors, etc. Exercise extreme care when using such equipment. Cool inoculating loops or needles before touching colonies on plates or inserting into liquid material.
6. Develop the habit of keeping your hands away from your mouth, nose, and eyes. Wash hands well with soap before leaving the laboratory.
7. Do not lay personal articles, such as eyeglasses, on the bench in your work area.
8. Laboratory coats or gowns are not to be worn outside the laboratory, particularly not to the employee lounge or cafeteria or out of the building.
9. Wipe off benches in your working area with disinfectant before and after each day’s work. Keep your area clean at all times.
10. In case of injury or unusual incident, however slight, the supervisor in charge must be immediately notified, and a report to occupational health facility is required. Also fill out an accident report form. Major accidents must be documented and reported in detail to the supervisor and chief of microbiology. The report should indicate:
A. Cause of the accident
B. Type of contamination or hazard
C. List of personnel possibly exposed and amount of exposure to possible pathogenic material
D. Decontamination procedures taken if pathogenic material was involved
E. Actions taken to prevent recurrence
F. Actions taken to safeguard or monitor employees
11. Infections may be spread by a number of routes in the laboratory. The actual occurrence of an infection depends on the virulence of the infecting agent, susceptibility of the host, route of entry, inoculating dose, etc. (Table 11.1).
A. Airborne. Droplets and aerosols may be formed by simply removing caps or cotton plugs or swabs from tubes. Heating liquids on needles too rapidly may also create an aerosol. Breakages in centrifuges are serious accidents (Table 11.2).
B. Ingestion. Ingestion can occur through mouth pipetting, failure to wash hands after handling specimens or cultures, or smoking, eating, and drinking in the laboratory.
C. Direct inoculation. Scratches, needlesticks, cuts from broken glass, or animal bites may permit direct inoculation.
D. Skin contact. Some very virulent organisms, and others not so virulent, can enter through small cuts or scratches or through the conjunctiva of the eye.
E. Vectors. Flies, mosquitoes, ticks, fleas, and other ectoparasites can be potential sources of infection in the laboratory, especially if animal work is performed.
12. Personnel who display risk-prone behavior or are pregnant, immunocompromised, or immunosuppressed should be restricted from performing work with highly infectious microorganisms and, in some situations, be restricted to a low-risk laboratory (22).
