Note on Streptococcus suis

S. Saengamnatdej, Ph.D.

October 2008
Features

• Alpha-hemolytic ; in viridans group (not pneumococci)

• Facultative anaerobe
• Gram positive
• Coccoid or ovoid (Spindle-shaped cocci in one article)
• Singly, pairs (frequently) or short chain (occasionally)
• 20 Mbp genome
• Thirty-five capsular serotypes (more in Epidemiology heading)
• Lancefield's group D
• Rebecca C. Lancefield (published in 1933)
• Classified hemolytic streptococci into five groups by precipitin test
• Prepare type-specific rabbit sera.
• Extract (water-clear supernatant fluid) from the neutralized HCl-treated
culture.
• Layer antiserum on the extract, stand tubes for 10-30 mins (RT/37C) and
observe ring formation, then shake and incubate at 37C for 2 hrs.
Finally, leave them sit on ice o/n before reading.
• Nowadays, it is performed by using Streptex (Remel, USA)

• The virulence differs among serotypes and between different strains of the same
serotype.

Swine diseases

• Disease in swine and others (boars, horses, dogs, cats, birds)

• Pigs can be carriers, in upper respiratory tract of pig (tonsils, nasal cavities), genital
and alimentary tract
• Serotype 2 is the most pathogenic to pigs (among the capsular serotype), and was
isolated in up to 50.6% of all S. suis isolates from healthy swine tonsil.

Occupational disease

• 1500 times higher risk in persons working in pork industry (Netherlands)

 • carriage rate 5.3% in high-risk group (Germany)
• 9% dairy farmers, 10% meat inspectors, 21% pig farmers (NZ)

Human diseases

• Meningitis (72.5%), others [septicaemia and septic shock (24.2%), arthritis,

endocarditis, spondylodiscitis (of spine disc), endophthamitis (of the intraocular
cavities), and uveitis (of the middle layer of the eye)]
• Hearing loss and vestibular dysfunction are marked sequelae
• High mortality rate (average of 17.8%), and higher if septicaemia.
• Recent outbreak (Sichuan, 2005) with high mortality rate, cause of dead (the cause??
toxic shock syndrome or new virulent factor)
• Symptom of fatal case similar to TSS (toxic-shock syndrome---patho=superantigen)
but No S. pyogenes toxin gene found.
• relapse---prolong treatment required

Physicians around the world should be aware of the possibility of S. suis-associated STSS
when they see patients with unexplained sepsis who had been in contact with pigs.

Epidemiology

• Zoonosis
• Unlikely human-to-human transmission without close contact with materials (blood)
• No obvious seasonal, but two Chinese outbreaks occurred in summer.
• first human (Denmark) case 1968
• Three outbreaks in China; in 1998, 1999, and 2005.
• spread worldwide, but three countries are comprised of almost 90% (China 69%,
Thailand 11.5% [with highest mortality rate26%] and Netherlands 8.3%)
• 2005 outbreak in china: 204 infected and 38 deaths
• Total 409 cases with 73 deaths (2007 review)
• Capsular serotypes: 35 types (1-34 and ½), but types 32 and 34 proven to be S.
orisratti.
• In most countries, capsular type 2 is currently the cause of most S. suis infections,
whereas in Denmark and Finland, capsular type 7 appears to be most prevalent.

Stability and Control

 • 60 °C for 10 mins.
• 50 °C for 2 hours.
• 10 °C for 6 weeks (carcasses).
• 0 °C for 1 month (in dust), for >3 months in faeces.
• 25 °C for 24 hours (in dust), for 8 days in faeces.
• Killed with 5% bleach (1:799 dilution)

Case report

• Taiwan Case 1 (38): fever, chill, headache, hearing impairment, acute OM, CSF
culture yielded S. suis. a bank clerk, no pig contact. (tx: ceftriaxone, then PenG)
• Taiwan case 2 (52): fever, nausea, headache, vomiting, neck stiffness, photophobia,
(tx: Amp + Ceftriaxone), blood culture: S. suis, sterile CSF, Skin itching (pen allergic →
ceftriaxone alone x14d), a pultry seller with a pig farm near her home.
• Taiwan case 3 (61), diarrhea, abd pain, consciousness change

Lab identification

• Culture
• rapid growth
• small colony (0.5-1.0 mm)
• gray-whitish colonies, or grayish or transparent and slightly mucoid.
• alpha-hemolysis (trypticase soy agar + 5% SRBC) 5%CO2, 35°C
• type 2 produces beta-hemolysis on horse blood agar plates.
• catalase-negative
• gram-positive cocci
• No growth in 6.5% NaCl media

• Biochemical test (Phenotypic reaction profiles)

• In an article, different test kits gave different results;
• S. suis by API Rapid ID20 Strep system (bioMerieux, Fr.) [biotype
profile, 0641473; identity, 99.8%],
• S. anginosus by Vitek GPI Card (bioMerieux Vitex, USA) [biotype profile,
5166333400; identity, 95%], and
• S. vestibularis by Phoenix System PID (Beckon Dickinson, USA)
[biotype profile, 420080163621; confidence value, 97%]
 • Species confirmed by
• 16S rRNA sequence analysis (1475 bp) and similarity search with BLAST : (in
one article found similarity with S. suis; AF009476; 99%)
• PCR
• rapid
• detect specific serotypes or strains of S. suis
• in animal carriers, infected or healthy pigs, and clinical samples.
• S. suis-specific 16S ribosomal RNA region
• a species-specific probe (serotypes 1-31) targeting 16S rRNA gene can be
used to identify S. suis strains.
• Multiplex PCR

• Capsular reaction (1990) = Quellung Reaction (used in S. pneumococci)

• Test of choice (type-specific serum + capsular polysaccharide material)
• 5-6 hr culture (to avoid aging)
• Easy, rapid, specific- technique
• A loopful of a 5%-serum culture broth is spread over an area of 0.5 cm in
diameter on a glass slide.
• One loopful of the antiserum is then mixed thoroughly with the culture
on the slide.
• Place a coverslip
• examined under a phase contrast microscope with 1000x magnification.
• Capsule visible (twice as big as those of control strains, which are mixed
with non-immune rabbit serum.)

• Suis Type Antisera (Statens Serum Institute, Copenhagen, Denmark) can be

used in the Quellung reaction.

• Other techniques
• immunocapture
• fluorescent antibody techniques
• whole-cell antigen-based indirect ELISA, and
• purified capsular polysaccharide antigen-based indirect ELISA.

Phylogenetic analysis and study of genetic diversity

• 16S rRNA gene

• chaperonin 60 gene
• RFLP
• genome fingerprint
• Pulsed-field gel electrophoresis (PFGE) with SmaI and ApaI
• Multilocus sequence typing (MLST)