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)