Bordetella Pertussis

Basic Characteristics

• Gram-negative (1)

• Rod-shaped, coccoid, or ovoid bacterium (2)

• Causes whooping cough

  • Highly contagious infection

• Found in mouth, nose, and respiratory tract (3)

History

Bordet and Gengou first discovered it in Paris in 1900. They identified “a small ovoid Gram-negative bacterium” in a specimen collected from a 5-month-old child. They actually isolated the bacterium 6 years later from Bordet’s own son. Since then, it has been responsible for multiple outbreaks with the largest peak in recent years in 2012. Furthermore, just this year there has been an outbreak in Nunavut community (4).

Domain: Bacteria

Phylum: Proteobacteria

Class: Betaproteobacteria

Order: Burkholderiales

Family: Alcaligenaceae

Genus: Bordetella

Species: B. pertussis

(https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi)

Pathogenicity

Bordetella pertussis is a pathogen that causes whooping cough. It does not have any benefits to the host (11).

It is important because it causes a highly contagious infection of the respiratory tract. The microbe is spread through respiratory droplets and the disease it causes results in coughing spasms that can contribute to spread. It can also cause severe harm to infants and can lead to many respiratory complications. Bordetella pertussis also has many self-defense mechanisms that help it persist longer in the respiratory track such as adenylate cyclase toxin which combats the body’s immune response, as well as pertussis toxin (3).

Antibiotics/Prevention

There is no proven effective treatment for reducing the pertussis symptoms, however antibiotics are used to prevent further transmission (3). Some antibiotics that are used are: Amoxicillin, Azithromycin, Clarithromycin, and Erythromycin. Additionally, babies get a vaccine when they are young to prevent infection.

(https://app.gideononline.com/explore/microbes/bacteria/800)

Safety

The BSL level for my microbe is 2. Basic laboratory measures such as wearing appropriate clothing and disinfecting work areas after use should be in place. Additionally, special containment practices and devices are generally required to decrease the risk to laboratory personnel.

Agar Type

You need a Bordet-Gengou Agar in order to grow Bordetella pertussis. This type of agar is optimized to isolate Bordetella. It is an enriched casein peptone medium and has a potato infusion as well as glycerol to supply nutrients. Blood is also added to supply additional nutrients and allow for the detection of hemolytic reactions.

Aerobic or Anaerobic conditions?

Bordetella pertussis needs to be grown in aerobic conditions.

Procedure for Growing

1. Open vial according to enclosed instructions

2. Use one tube of #35 broth (5-6 mL)

3. Use pipette to withdraw 0.5-1 mL and rehydrate entire pellet

4. Aseptically transfer this back into the broth tube.

5. Mix well

6. Incubate the tubes and plate at 37 degrees Celsius for 24-48 hours

(https://www.atcc.org/products/all/8467.aspx#documentation)

Using primers in literature to run in In silico PCR amplification and BLAST to find 16S ribosomal RNA

Primers:

• 16S-F2: 5’-AACTGCATTTTTAACTACCGG-3’ (12)

  • Melting temperature: 48.5 Celsius

  • GC content: 38%

• 16S-R1: 5'-ATCCTGTTTGCTCCCCACA-3' (12)

  • Melting temperature: 51.1 Celsius

  • GC content: 53%

(http://biotools.nubic.northwestern.edu/OligoCalc.html)

In silico PCR Amplification result:

>NC_002929, from 2149665 to 2149824 (160 bp); Bordetella pertussis

AACTGCATTTTTAACTACCGGGCTAGAGTGTGTCAGAGGGAGGTGGAATTCCGCGTGTAGCAGTGAAATGCGTAGATATGCGGAGGAACACCGATGGCGAAGGCAGCCTCCTGGGATAACACTGACGCTCATGCACGAAAGTGTGGGGAGCAAACAGGAT

(http://insilico.ehu.eus/find_gene.php?genome=002929&start=2149665&length=160)

BLAST Result:

>NR_025951.1 Bordetella pertussis strain 18323 16S ribosomal RNA, partial sequence

AACTGAAGAGTTTGATCCTGGCTCAGATTGAACGCTGGCGGGATGCTTTACACATGCAAGTCGGACGGCAGCACGGGCTTCGGCCTNGTGGCGAGTGGCGAACGGGTGAGTAATGTATCGGAACGTGCCCAGTAGCGGGGGATAACTACGCGAAAGCGTAGCTAATACCGCATACGCCCTACGGGGGAAAGCGGGGGACCTTCGGGCCTCGCACTATTGGAGCGGCCGATATCGGATTAGCTNGTTGGTGGGGTAACGGCCTACCAAGGCGACGATCCGTAGCTGGTTTGAGAGGACGACCAGCCACACTGGGACTGAGACACGGCCCAGNCTCCTACGGGAGGCAGCAGTGGGGAATTTTGGACAATGGGGGCAACCCTGATCCAGCCATCCCGCGTGTGCGATGAAGGCCTTCGGGTTGTAAAGCACTTTTGGCAGGAAAGAAACGGCACGGGCTAATATCCTGTGCAACTGACGGTACCTGCAGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTTCGGAAAGAAAGATGTGAAATCCCAGGGCTTAACCTTGGAACTGCATTTTTAACTACCGGGCTAGAGTGTGTCAGAGGGAGGTGGAATTCCGCGTGTAGCAGTGAAATGCGTAGATATGCGGAGGAACACCGATGGCGAAGGCAGCCTCCTGGGATAACACTGACGCTCATGCACGAAAGTGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCCTAAACGATGTCAACTAGCTGTTGGGGCCTTCGGGCCTTGGTAGCGCAGCTAACGCGTGAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCAACGCGAAAAACCTTACCTACCCTTGACATGTCTGGAATCCCGAAGAGATTTGGGAGTGCTCGCAAGAGAACCGGAACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCATTAGTTGCTACGAAAGGGCACTCTAATGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTGAAGTCCTCATGGCCCTTATGGGTAGGGCTTCACACGTCATACAATGGTCGGGACAGAGGGTTGNCAACCCGCGAGGGGGAGCCAATCCCAGAAACCCGGTCGTNGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGTCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCNCGTCACACCATGGGAGTGGGTTTTACCAGAAGTAGTTAGCCTAACCGCAAGGGGGGCGATTACCACGGTAGGA

(https://blast.ncbi.nlm.nih.gov/Blast.cgi#alnHdr_219846360)