Pathogen Safety Data Sheets:

Infectious Substances –
Corynebacterium diphtheriae

PATHOGEN SAFETY DATA SHEET -

INFECTIOUS SUBSTANCES

SECTION I - INFECTIOUS AGENT

NAME: Corynebacterium diphtheriae
SYNONYM OR CROSS REFERENCE: Diphtheria Footnote1.
CHARACTERISTICS: Corynebacterium diphtheriae is part of
the Corynebacteriaceae family and genus Corynebacterium. Bacteria are small,
pleomorphic, aerobic, non-spore forming bacilli. They are Gram positive and
slightly club shaped Footnote1, Footnote2. C. diphtheriae cells can be arranged as single
cells, in pairs, in V forms, in palisades, or in clusters Footnote2. They are non-motile
and catalase positive Footnote2. Non-hemolytic colonies appear within 18-24 hours
when cultured on media containing blood at 37°C Footnote1, Footnote2. Biotypes (or
biovars) of C. diphtheriae are gravis, mitis, belfanti and intermedius Footnote1.

SECTION II - HAZARD IDENTIFICATION

PATHOGENICITY/TOXICITY: C. diphtheriae is a causative agent of diphtheria
(respiratory tract/pharyngreal/throat diphtheria or cutaneous diphtheria) Footnote2.
Respiratory tract diphtheria: An upper respiratory tract illness which manifests as
pharyngitis or tonsillitis with sore throat, dysphagia, lymphadenitis, low grade
fever, malaise, and headache Footnote1, Footnote2. In severe cases, severe adenitis and
soft tissue edema may result in bull neck appearance Footnote3. A patch of
membrane develops on the tonsils which may extend to tonsillar pillars, uvula,
soft palate, oropharynx, and nasopharynx Footnote1, Footnote3. Edema and
pseudomembrane coating of the trachea and bronchi can also occur Footnote3. In
uncomplicated cases, the disease usually resolves and the membrane is
coughed up after 5-10 days Footnote1. Serious effects of diphtheria such as
myocarditis, neuritis, and kidney damage are caused by systemic absorption of
the diphtheria exotoxin from the site of infection Footnote1, Footnote2, Footnote4. Myocarditis is
manifested by cardiac enlargement and weakness, arrhythmia and congestive
heart failure Footnote1. About 75% of patients with severe respiratory diphtheria may
also develop neuropathy Footnote3. Infection of the nervous system causes paralysis
of the soft palate, or oculomotor muscles, which usually resolves with the
formation of antitoxin antibody Footnote1. Serious and lethal effects of diphtheria may
also ensue from mechanical obstruction of the airway due to pseudomembrane
extension into the trachea and bronchi, resulting in cyanosis and suffocation of
the patient Footnote1, Footnote3.
Cutaneous diphtheria is most common in tropical, hot, and arid regions Footnote4. It is
characterized by formation of lesions on the skin, which may range from a simple
pustule to a chronic non-recovering ulcer Footnote1, Footnote2, Footnote3. C. diphtheriae mainly
colonizes preexisting lesions on the skin such as surgical wounds, pyoderma,
eczema, impetigo, dermatitis, or insect bites Footnote3. Corynebacterium
ulcerans and C. pseudotuberculosis can also harbour diphtheria tox genes and
express diphtheria toxin. Infections by these species most often occur in animals
but can also cause diphtheria-like disease in humans, with symptoms similar to
those described here for C. diphtheriae. Of increasing concern are infections by
toxigenic strains of C. ulcerans found in companion pets such as dogs or cats,
which subsequently can cause zoonotic infections in humans Footnote1.
EPIDEMIOLOGY: The WHO reported 7088 cases of diphtheria world wide in
2008 and 5000 estimated deaths in 2004 Footnote5. Due to routine immunization
programs, diphtheria is rare in developed countries, although, cases that are
observed tend to be more serious, and may be fatal Footnote4. In these countries,
diphtheria usually occurs as small outbreaks in the unimmunized population such
as among migrant workers or transients Footnote1. In Canada, diphtheria has been
limited to mainly small outbreaks in First Nations (i.e., indigenous North American
Indian) populations, with the most recent outbreaks occurring over 30 years
ago Footnote6. Despite the widespread use of immunization, diphtheria remains
endemic in developing countries such as India, Bangladesh, Vietnam, countries
in Africa and areas of South America (i.e. Brazil) Footnote7. In Russia, diphtheria
remerged in 1990 and spread to all the other countries that previously formed the
Soviet Union, as well as the Baltic States. The annual number of diphtheria cases
was below 200 before 1990, and increased to 47,000 with 1700 deaths between
1990 and 1995, and approximately157,000 cases with 5000 deaths by the
beginning of 1999 Footnote1, Footnote7.
HOST RANGE:Humans. Although rare, some biotypes of C. diphtheriae have
also been isolated from animals such as cow, cats, and horses Footnote8. C.
ulceransand C. pseudotuberculosis are derived from animals but also cause
disease in humans Footnote1.
INFECTIOUS DOSE: Unknown; however, the main virulence factor of
Corynebacterium diphtheriae, C. ulcerans and C. pseudotuberculosis is
diphtheria toxin, which has a lethal dose of < 0.1 µg/kg of body weight in humans
Footnote 4, Footnote 9.
MODE OF TRANSMISSION: Transmission of C. diphtheriae can occur through
droplet nuclei, fomites, and direct contact with cutaneous infections Footnote1. C.
diphtheriae has also been transmitted from hospital wards and clothing Footnote10.
Diphtheria has also been transmitted through the consumption of milk Footnote11.
Transmission of C. ulcerans and C. pseudotuberculosisis are thought to be
similar to that of C. diphtheriae Footnote1.
INCUBATION PERIOD: 2-4 days Footnote1
COMMUNICABILITY: Variable period. In the absence of antibiotic therapy,
disease is communicable for 2-4 weeks Footnote10. Appropriate antibiotic therapy
usually terminates bacterial shedding within 48 hours Footnote10.

SECTION III - DISSEMINATION

RESERVOIR: Humans are thought to be a significant reservoir for C.
diphtheriae Footnote10. C. diphtheriae may be present in nasopharynx and skin lesions
for weeks to months or even for life time in asymptomatic individuals, which can
act as a reservoir for the disease Footnote1, Footnote2. The reservoirs for C.
ulcerans and C. pseudotuberculosis are thought to be primarily those of
animals Footnote1.
ZOONOSIS: None
VECTORS: None

SECTION IV - STABILITY AND VIABILITY

DRUG SUSCEPTIBILITY/ RESISTANCE: Susceptible to penicillin, erythromycin,
tetracycline and certain cephalosporins Footnote1, Footnote12. Some strains resistant to
erythromycin have been reported Footnote12.
SUSCEPTIBILITY/RESISTANCE TO DISINFECTANTS: C. diphtheriae and
related species can also be inhibited by chlorhexidine (MIC of 5mg/l) Footnote13. Most
vegetative bacteria are also susceptible to 1% sodium hypochlorite, 70% ethanol,
glutaraldehyde, formaldehyde, iodines, hydrogen peroxide, peracetic acid, and
quaternary ammonium compounds Footnote14.
PHYSICAL INACTIVATION: Information specific to C. diphtheriae and related
species is not available, but most vegetative bacteria can be inactivated by moist
heat (121°C for 15 min- 30 min) and dry heat (160-170°C for 1-2 hours) Footnote15.
SURVIVAL OUTSIDE HOST: C. diphtheriae can survive on dry inanimate
surfaces from 7 days to 6 months Footnote16. Survival of C. ulcerans and C.
pseudotuberculosis is unknown.

SECTION V – FIRST AID / MEDICAL

SURVEILLANCE: Monitor for symptoms. Diagnosis of diphtheria is done mainly
through monitoring of clinical symptoms (lesions on the skin or formation of
pseudomembrane) Footnote1, Footnote2. Confirmation of C. diphtheriae infection can be
done by culture of the bacteria on selective media such as 5% sheep blood agar,
tinsdale medium, and colistin-nalidixic acid blood agar, followed by Gram
staining, and differential biochemical tests Footnote1, Footnote2. Other methods for
identification of these species include: gas-liquid chromatography of cellular fatty
acids, and increasingly, the use of 16SrRNA and rpoB gene sequencing Footnote2.
Note: All diagnostic methods are not necessarily available in all countries.
FIRST AID/TREATMENT: Administration of diphtheria antitoxin (DAT) is the most
successful treatment for diphtheria Footnote4. In order to prevent further toxin
production, antibiotic therapy with penicillins, cephalosporins, erythromycin, and
tetracycline may be used in conjunction with antitoxin to eliminate the bacteria
from the site of infection Footnote1, Footnote4, Footnote10. Penicillin can be given intramuscularly
or orally Footnote10.
IMMUNIZATION: Vaccination for the control of diphtheria infection is given as a
combined DTaP vaccine of Diphtheria, pertussis, and tetanus toxoid Footnote17, Footnote18.
It is administered along with the vaccine for poliomyelitis and Haemophilus
influenzae type b (DTaP-IPV-Hib) at 2, 4, 6 and 18 months of age Footnote18, Footnote19. A
booster dose of DTaP–IPV vaccine is also given between 4 and 6 years of
age Footnote18. A single booster dose of dTpa (Boostrix TM) can be also be used to
prevent diphtheria, tetanus, and pertussis in individuals more than 4 years of age
in Europe and Canada, in adolescents aged 10–18 years in the US, and in
individuals over the age of 10 in Australia Footnote17. This vaccine contains the same
toxoid as DTap vaccine, and is used for primary immunization, but in reduced
quantities to prevent increased immunological reactions with consecutive
doses Footnote17. Individuals should be boosted every 10 years after conclusion of
childhood vaccination regime Footnote20.
PROPHYLAXIS: Antibiotic prophylaxis with intramuscular benzathine penicillin G
is recommended for close household contacts and medical staff exposed to oral
secretions of the infected individual. Oral erythromycin can also be used Footnote10.

SECTION VI - LABORATORY HAZARDS

LABORATORY-ACQUIRED INFECTIONS: Thirty-three cases of laboratory
acquired diphtheria were reported as of 1976 Footnote21.
SOURCE/SPECIMENS: Exudates or secretions from nose, throat, nasopharynx,
larynx, wounds, blood, skin Footnote2, Footnote22.
PRIMARY HAZARDS: Inhalation, accidental parenteral inoculation, and
ingestion Footnote22.
SPECIAL HAZARDS: None.

SECTION VII – EXPOSURE CONTROLS / PERSONAL

PROTECTION
RISK GROUP CLASSIFICATION: Risk Group 2 Footnote23.
CONTAINMENT REQUIREMENTS: Containment Level 2 facilities, equipment,
and operational practices for work involving infected or potentially infected
materials, animals, or cultures Footnote24.
PROTECTIVE CLOTHING: Lab coat. Gloves when direct skin contact with
infected materials or animals is unavoidable. Eye protection must be used where
there is a known or potential risk of exposure to slashes Footnote24.
OTHER PRECAUTIONS: All procedures that may produce aerosols, or involve
high concentrations or large volumes should be conducted in a biological safety
cabinet (BSC). The use of needles, syringes, and other sharp objects should be
strictly limited. Additional precautions should be considered with work involving
animals or large scale activities Footnote24.
SECTION VIII – HANDLING AND STORAGE
SPILLS: Allow aerosols to settle and, wearing protective clothing, gently cover
spill with paper towels and apply an appropriate disinfectant, starting at the
perimeter and working towards the centre. Allow sufficient contact time before
clean up Footnote24.
DISPOSAL: Decontaminate all wastes that contain or have come in contact with
the infectious organism by autoclave, chemical disinfection, gamma irradiation, or
incineration before disposing Footnote24.
STORAGE: The infectious agent should be stored in leak-proof containers that
are appropriately labeled Footnote24.

SECTION IX - REGULATORY AND OTHER

INFORMATION
REGULATORY INFORMATION: The import, transport, and use of pathogens in
Canada is regulated under many regulatory bodies, including the Public Health
Agency of Canada, Health Canada, Canadian Food Inspection Agency,
Environment Canada, and Transport Canada. Users are responsible for ensuring
they are compliant with all relevant acts, regulations, guidelines, and standards.
UPDATED: December 2011
PREPARED BY: Pathogen Regulation Directorate, Public Health Agency of
Canada
Although the information, opinions and recommendations contained in this
Pathogen Safety Data sheet are compiled from sources believed to be reliable,
we accept no responsibility for the accuracy, sufficiency, or reliability or for any
loss or injury resulting from the use of the information. Newly discovered hazards
are frequent and this information may not be completely up to date.
Copyright ©
Public Health Agency of Canada, 2011
Canada

REFERENCES:
Footnote 1
Ryan, K. J. (2004). Corynebacterium, Listeria, and Bacillus. In K.
J. Ryan, & C. G. Ray (Eds.), Sherris Medical Microbiology: An
Introduction to Infectious Diseases (4th ed., pp. 297-308). USA:
McGraw Hill.

Return to footnote1Referrer
Footnote 2
Funke, G. and K. Bernard. (2007). Coryneform Gram-Positive
Rods. In P. R. Murray (Ed.), Manual of Clinical Microbiology (9th
 ed., pp. 485-514). Washington D.C.: ASM Press.

Return to footnote2Referrer
Footnote 3
Hadfield, T. L., McEvoy, P., Polotsky, Y., Tzinserling, V. A., &
Yakovlev, A. A. (2000). The pathology of diphtheria. Journal of
Infectious Diseases, 181(Suppl 1), S116-20.

Return to footnote3Referrer
Footnote 4
Wagner, K. S., Stickings, P., White, J. M., Neal, S., Crowcroft, N.
S., Sesardic, D., & Efstratiou, A. (2009). A review of the
international issues surrounding the availability and demand for
diphtheria antitoxin for therapeutic use. Vaccine, 28(1), 14-20.

Return to footnote4Referrer
Footnote 5
World Health Organization. (2010). Immunization, Surveillance,
Assessment and Monitoring. Retrieved APRIL 7, 2010, from
http://www.who.int/immunization_monitoring/diseases/diphteria/en
/

Return to footnote5Referrer
Footnote 6
Romney, M. G., Roscoe, D. L., Bernard, K., Lai, S., Efstratiou, A.,
& Clarke, A. M. (2006). Emergence of an invasive clone of
nontoxigenic Corynebacterium diphtheriae in the urban poor
population of Vancouver, Canada. Journal of Clinical Microbiology,
44(5), 1625-1629. doi:10.1128/JCM.44.5.1625-1629.2006

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Footnote 7
Mattos-Guaraldi, A. L., Moreira, L. O., Damasco, P. V., & Hirata
Junior, R. (2003). Diphtheria remains a threat to health in the
developing world--an overview. Memorias do Instituto Oswaldo
Cruz, 98(8), 987-993.

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Footnote 8
Hall, A. J., Cassiday, P. K., Bernard, K. A., Bolt, F., Steigerwalt, A.
G., Bixler, D., Pawloski, L. C., Whitney, A. M., Iwaki, M., Baldwin,
A., Dowson, C. G., Komiya, T., Takahashi, M., Hinrikson, H. P., &
Tondella, M. L. (2010). Novel Corynebacterium diphtheriae in
domestic cats. Emerging Infectious Diseases, 16(4), 688-691.

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Footnote 9
 Spinler, J. K., & Holmes, R. K. (2004). Corynebacterium
diphtheriae—Molecular Detection of Diphtheria
Toxin. Encyclopedia of Medical Genomics and Proteomics, , 297.
Retrieved from http://www.informaworld.com/10.1081/E-EDGP-
120024073

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Footnote 10
Begg, N., & Balraj, V. (1995). Diphtheria: are we ready for
it? Archives of Disease in Childhood, 73(6), 568-572.

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Footnote 11
Barrett, N. J. (1986). Communicable disease associated with milk
and dairy products in England and Wales: 1983-1984. The Journal
of Infection, 12(3), 265-272.

Return to footnote11Referrer
Footnote 12
Kneen, R., Pham, N. G., Solomon, T., Tran, T. M., Nguyen, T. T.,
Tran, B. L., Wain, J., Day, N. P., Tran, T. H., Parry, C. M., &
White, N. J. (1998). Penicillin vs. erythromycin in the treatment of
diphtheria. Clinical Infectious Diseases, 27(4), 845-850.

Return to footnote12Referrer
Footnote 13
Larsson, P., Brinkhoff, B., & Larsson, L. (1987). Corynebacterium
diphtheriae in the environment of carriers and patients. Journal of
Hospital Infection, 10(3), 282-286.

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Footnote 14
Rutala, W. A. (1996). APIC guideline for selection and use of
disinfectants. American Journal of Infection Control, 24(4), 313-
342.

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Footnote 15
Pflug, I. J., Holcomb, R. G., & Gomez, M. M. (2001). Principles of
the thermal destruction of microorganisms. In S. S. Block
(Ed.), Disinfection, Sterilization, and Preservation (5th ed., pp. 79-
129). Philadelphia, PA: Lipincott Williams and Wilkins.

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Footnote 16
Kramer, A., Schwebke, I., & Kampf, G. (2006). How long do
nosocomial pathogens persist on inanimate surfaces? A
 systematic review. BMC Infectious Diseases, 6

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Footnote 17
Frampton, J. E., & Keating, G. M. (2006). Reduced-antigen,
combined diphtheria, tetanus, and acellular pertussis vaccine
(Boostrix): a review of its use as a single-dose booster
immunization. Biodrugs, 20(6), 371-389.

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Footnote 18
Galanis, E., King, A. S., Varughese, P., & Halperin, S. A. (2006).
Changing epidemiology and emerging risk groups for
pertussis. Canadian Medical Association Journal, 174(4), 451-
452.

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Footnote 19
Greenberg, D. P., Doemland, M., Bettinger, J. A., Scheifele, D. W.,
Halperin, S. A., Waters, V., & Kandola, K. (2009). Epidemiology of
pertussis and haemophilus influenzae type b disease in Canada
with exclusive use of a diphtheria-tetanus-acellular pertussis-
inactivated poliovirus-haemophilus influenzae type b pediatric
combination vaccine and an adolescent-adult tetanus-diphtheria-
acellular pertussis vaccine: Implications for disease prevention in
the United States. Pediatric Infectious Disease Journal, 28(6),
521-528.

Return to footnote19Referrer
Footnote 20
Public Health Agency of Canada. (2006). Canadian Immunization
Guide.Seventh Edition - 2006

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Footnote 21
Pike, R. M. (1976). Laboratory associated infections: summary
and analysis of 3921 cases. Health Laboratory Science, 13(2),
105-114.

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Footnote 22
Agent Summary Statements:Bacterial Agents. (1999). In J. Y.
Richmond, & R. W. Mckinney (Eds.), Biosafety in Microbiological
and Biomedical Laboratories (BMBL) (4th ed., pp. 88-117).
Washington, D.C.: Centres for Disease Control and Prevention.

Return to footnote22Referrer
Footnote 23
 Human Pathogens and Toxins Act. S.C. 2009, c. 24. Government
of Canada, Second Session, Fortieth Parliament, 57-58 Elizabeth
II, 2009, (2009).

Return to footnote23Referrer
Footnote 24
Public Health Agency of Canada. (2004). In Best M., Graham M.
L., Leitner R., Ouellette M. and Ugwu K. (Eds.), Laboratory
Biosafety Guidelines (3rd ed.). Canada: Public Health Agency of
Canada.