Streptococcus pneumoniae

Characteristics
Morphology	S. pneumoniae is of the Streptococcaceae family. The gram-positive, oval/lancet-shaped cocci are often arranged in pairs, known as a diplococcus, or can be present in short chains. There are around 90 serotypes and its surface capsule, which is the distinguishing trait of the pneumococcus and is the major virulent factor, holds a complex mosaic of monosaccharides, oligosaccharides, and other polymer components that are of high-molecular weight
Growth Conditions	Cultures can be placed in various growth media and incubated at 37°C. When grown on blood agar, it forms round facultative anaerobic colonies surrounded by α-hemolysis.

Characteristics

Morphology

S. pneumoniae is of the Streptococcaceae family. The gram-positive, oval/lancet-shaped cocci are often arranged in pairs, known as a diplococcus, or can be present in short chains. There are around 90 serotypes and its surface capsule, which is the distinguishing trait of the pneumococcus and is the major virulent factor, holds a complex mosaic of monosaccharides, oligosaccharides, and other polymer components that are of high-molecular weight

Growth
Conditions

Cultures can be placed in various growth media and incubated at 37°C. When grown on blood agar, it forms round facultative anaerobic colonies surrounded by α-hemolysis.

Health Hazards
Host Range	Humans, mice, rats, guinea pigs, chimpanzees, rhesus monkeys, and other mammals associated with humans
Modes of Transmission	Infectious cells can be disseminated via microaerosol droplets created by coughing or sneezing, or person-person oral contact. Transmission is common, but infection is infrequent as healthy individuals carry S. pneumoniae in the nasopharyngeal region without any presence of infection. Pneumococcus bacteria can cause infections in many parts of the body including: lungs, ears, sinuses, brain, and spinal tissue
Signs and Symptoms	Symptoms of pneumococcal infection depend on the part of the body affected. Symptoms can include fever, cough, shortness of breath, chest pain, stiff neck, confusion, increased sensitivity to light, joint pain, chills, ear pain, sleeplessness, and irritability. In severe cases, pneumococcal disease can cause hearing loss, brain damage, sepsis, and death.
Infectious Dose	Unknown
Incubation Period	About 1 – 3 days upon breaching primary immune barriers

Medical Precautions/Treatment
Prophylaxis	Antibiotic prophylaxis is available
Vaccines	Vaccines are available, Pneumococcal conjugate vaccine (PCV13) and pneumococcal polysaccharide vaccine (PPV23)
Treatment	Administer appropriate drug treatment. Inflammation caused by pneumococcal lysis makes the treatment of pneumococcal diseases less effective with antibiotics alone, and even a highly effective bactericidal such as β-lactam may actually enhance the harmful effects of the disease in some cases
Surveillance	Monitor for symptoms. Gram-negative stains or bacteriology studies of direct smears can be used to detect symptomatic or asymptomatic infections. ELISA, PCR fingerprinting analysis, and radiography techniques are also useful for diagnosis.
GWU Requirements	Report all incidents to the Office of Risk Management as well as the Office of Research Safety (ORS) IMMEDIATELY FOLLOWING THE OCCURRENCE.

Containment
BSL-2/ABSL-2	Risk Group 2 classification is applied VC. BSL-2 practices, containment equipment, and facilities are recommended for activities with cultures or potentially infectious clinical materials. ABSL-2 practices, containment equipment, and facilities are recommended for activities with naturally or experimentally infected animals.

Spill Procedures
Small Spills (<1 liter)	If the spill occurred inside a biological safety cabinet, close the sash and allow the cabinet to operate for 15 minutes before continuing with the spill cleanup. S. pneumoniae is not commonly transmitted via aerosols, but caution should be taken during a spill outside of a BSC. Leave the room immediately and allow the aerosols to dissipate for 15 minutes. Notify others working in the lab. Don appropriate PPE. Cover area of the spill with paper towels or any absorbent material and apply an EPA registered disinfectant effective against the bacteria (bleach, ethanol, glutaraldehyde), working from the perimeter towards the center. Allow 30 minutes of contact time before disposal and cleanup of spill materials.
Large Spills	Alert lab personnel in the laboratory to the spill and keep people out of the area to prevent spread of the contamination. Check if you have been contaminated or if any of your PPE has been breached. If so follow exposure procedures. Remove any contaminated clothing and place it the biohazard waste. Wash your hands and post a sign on the door. Notify your supervisor of the incident and call ORS (4-8258) for assistance. If the situation involves an imminently life-threatening injury or has catastrophic potential, call 911.

Spill Procedures

Small Spills (<1 liter)

If the spill occurred inside a biological safety cabinet, close the sash and allow the cabinet to operate for 15 minutes before continuing with the spill cleanup.

S. pneumoniae is not commonly transmitted via aerosols, but caution should be taken during a spill outside of a BSC. Leave the room immediately and allow the aerosols to dissipate for 15 minutes. Notify others working in the lab. Don appropriate PPE. Cover area of the spill with paper towels or any absorbent material and apply an EPA registered disinfectant effective against the bacteria (bleach, ethanol, glutaraldehyde), working from the perimeter towards the center. Allow 30 minutes of contact time before disposal and cleanup of spill materials.

Large Spills

Alert lab personnel in the laboratory to the spill and keep people out of the area to prevent spread of the contamination. Check if you have been contaminated or if any of your PPE has been breached. If so follow exposure procedures. Remove any contaminated clothing and place it the biohazard waste. Wash your hands and post a sign on the door. Notify your supervisor of the incident and call ORS (4-8258) for assistance. If the situation involves an imminently life-threatening injury or has catastrophic potential, call 911.

Exposure Procedures
Mucus Membrane	Flush eyes, mouth or nose for 15 minutes at eyewash station.
Other Exposure	For an area not protected by skin, wash with soap and water for 15 minutes (open wounds, sores, etc.)
Reporting	Report ALL injuries to the PI immediately and reported to the Office of Risk Management at [email protected] IMMEDIATELY FOLLOWING THE OCCURRENCE. Exposures that involve a bloodborne-pathogen or recombinant DNA also need to be reported to the Office of Research Safety at [email protected]. If the injury requires immediate medical attention, call GWPD at 202-994-6111 or call 911.
Medical Monitoring	Seek immediate medical evaluation, treatment, and post exposure follow-up at the Employee Health Office at GWU Hospital (900 23rd St., NW, Suite G-1090, Phone: 202-715-4275). Students should go to the Students Health Office at Marvin Center. After hours treatment can be received at the GWU hospital emergency room.

Stability
Disinfection	Susceptible to 10% bleach, 0.5% glutaraldehyde, 70% ethanol
Inactivation	Cells can be inactivated by heat suspension in a water bath at 56˚C for 30 minutes
Survival Outside Host	Can survive in dental plaque and sputum for up to 7 days, in dust for up to 20 days, on glass for 1 – 11 days, up to 180 days in frozen fish, and on mouse carcasses for up to 180 to 270 days.

Personal Protective Equipment (PPE)
Minimum PPE Requirements	At minimum, personnel are required to don gloves, closed toed shoes, lab coat, and appropriate face and eye protection prior to working with S. pneumoniae. Additional PPE may be required depending on lab specific SOPs.
Additional Precautions	Additional protection may be worn over laboratory clothing when infectious materials are directly handled, such as solid-front gowns with tight fitting wrists, gloves, and respiratory protection. Wash hands with soap and water after removing gloves.

References
CDC (https://www.cdc.gov/pneumococcal/index.html) BMBL 6th Edition (https://www.cdc.gov/labs/BMBL.html) Canadian PHAC (https://www.canada.ca/en/public-health/services/laboratory-biosafety-bi…)