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Construction and characterization of a Streptococcus suis serotype 2 recombinant expressing enhanced green fluorescent protein.

Chen T, Huang Q, Li Z, Zhang W, Lu C, Yao H - PLoS ONE (2012)

Bottom Line: We did not find significant differences in lethality (50% lethal dose), morbidity and mortality between the two strains.Our results also showed that the Egfp-HA9801 cells grown at 37°C for 36 h displayed greater green fluorescence signals than the cells grown at 28°C for 36 h and 37°C for 24 h.Together, these results indicate that the egfp and spc(r) insertions into the Egfp-HA9801 recombinant did not significantly change the virulence when compared with HA980, and this EGFP labeled strain can be used for future S. suis 2 pathogenesis research.

View Article: PubMed Central - PubMed

Affiliation: Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, China.

ABSTRACT
Streptococcus suis serotype 2 (S. suis 2) is an important pathogen, responsible for diverse diseases in swine and humans. To obtain a S. suis 2 strain that can be tracked in vitro and in vivo, we constructed the Egfp-HA9801 recombinant S. suis 2 strain with egfp and spc(r) genes inserted via homologous recombination. To assess the effects of the egfp and spc(r) genes in HA9801, the biochemical characteristics, growth features and virulence in Balb/C mice were compared between the recombinant and the parent HA9801 strain. We detected the EGFP expression from Egfp-HA9801 by epifluorescence microscopy. The results showed that the biochemical characterization and growth features of the Egfp-HA9801 recombinant were highly similar to that of the parent HA9801. We did not find significant differences in lethality (50% lethal dose), morbidity and mortality between the two strains. Furthermore, the bacterial counts in each various tissues of Egfp-HA9801-infected mice displayed similar dynamic compared with the HA9801-infected mice. Our results also showed that the Egfp-HA9801 cells grown at 37°C for 36 h displayed greater green fluorescence signals than the cells grown at 28°C for 36 h and 37°C for 24 h. The fluorescence in the tissue cryosections of Egfp-HA9801-injected mice was also stronger than that of the HA9801 group. Together, these results indicate that the egfp and spc(r) insertions into the Egfp-HA9801 recombinant did not significantly change the virulence when compared with HA980, and this EGFP labeled strain can be used for future S. suis 2 pathogenesis research.

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Survival curves for Balb/C mice infected with Egfp-HA9801 and HA9801strains.Six-week-old Balb/C mice were inoculated i.p. with 6.0×108 CFU bacteria; mice survival was monitored over a 7-day period. Data are expressed as mean percentage of live animals in each group (n = 10).
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pone-0039697-g003: Survival curves for Balb/C mice infected with Egfp-HA9801 and HA9801strains.Six-week-old Balb/C mice were inoculated i.p. with 6.0×108 CFU bacteria; mice survival was monitored over a 7-day period. Data are expressed as mean percentage of live animals in each group (n = 10).

Mentions: During the course of infection, all the mice infected with HA9801 at approximately 6.0×108 CFU showed clear clinical signs, such as depression, swollen eyes, weakness and prostration post-inoculation. Five mice died in this group during the 24 h post-inoculation. From day 4 p.i., surviving mice in the HA9801 group developed clinical signs associated with S. suis 2 meningitis such as hyperexcitation, opisthotonus, bending of the head and walking in circles. Meanwhile, mice infected with Egfp-HA9801 had an 80% mortality rate, and all of them showed clear clinical signs (Table 5 and Fig. 3). All the results indicated that no significant differences were found between the Egfp-HA9801 group and the HA9801 group.


Construction and characterization of a Streptococcus suis serotype 2 recombinant expressing enhanced green fluorescent protein.

Chen T, Huang Q, Li Z, Zhang W, Lu C, Yao H - PLoS ONE (2012)

Survival curves for Balb/C mice infected with Egfp-HA9801 and HA9801strains.Six-week-old Balb/C mice were inoculated i.p. with 6.0×108 CFU bacteria; mice survival was monitored over a 7-day period. Data are expressed as mean percentage of live animals in each group (n = 10).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3401235&req=5

pone-0039697-g003: Survival curves for Balb/C mice infected with Egfp-HA9801 and HA9801strains.Six-week-old Balb/C mice were inoculated i.p. with 6.0×108 CFU bacteria; mice survival was monitored over a 7-day period. Data are expressed as mean percentage of live animals in each group (n = 10).
Mentions: During the course of infection, all the mice infected with HA9801 at approximately 6.0×108 CFU showed clear clinical signs, such as depression, swollen eyes, weakness and prostration post-inoculation. Five mice died in this group during the 24 h post-inoculation. From day 4 p.i., surviving mice in the HA9801 group developed clinical signs associated with S. suis 2 meningitis such as hyperexcitation, opisthotonus, bending of the head and walking in circles. Meanwhile, mice infected with Egfp-HA9801 had an 80% mortality rate, and all of them showed clear clinical signs (Table 5 and Fig. 3). All the results indicated that no significant differences were found between the Egfp-HA9801 group and the HA9801 group.

Bottom Line: We did not find significant differences in lethality (50% lethal dose), morbidity and mortality between the two strains.Our results also showed that the Egfp-HA9801 cells grown at 37°C for 36 h displayed greater green fluorescence signals than the cells grown at 28°C for 36 h and 37°C for 24 h.Together, these results indicate that the egfp and spc(r) insertions into the Egfp-HA9801 recombinant did not significantly change the virulence when compared with HA980, and this EGFP labeled strain can be used for future S. suis 2 pathogenesis research.

View Article: PubMed Central - PubMed

Affiliation: Key Lab of Animal Bacteriology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing, China.

ABSTRACT
Streptococcus suis serotype 2 (S. suis 2) is an important pathogen, responsible for diverse diseases in swine and humans. To obtain a S. suis 2 strain that can be tracked in vitro and in vivo, we constructed the Egfp-HA9801 recombinant S. suis 2 strain with egfp and spc(r) genes inserted via homologous recombination. To assess the effects of the egfp and spc(r) genes in HA9801, the biochemical characteristics, growth features and virulence in Balb/C mice were compared between the recombinant and the parent HA9801 strain. We detected the EGFP expression from Egfp-HA9801 by epifluorescence microscopy. The results showed that the biochemical characterization and growth features of the Egfp-HA9801 recombinant were highly similar to that of the parent HA9801. We did not find significant differences in lethality (50% lethal dose), morbidity and mortality between the two strains. Furthermore, the bacterial counts in each various tissues of Egfp-HA9801-infected mice displayed similar dynamic compared with the HA9801-infected mice. Our results also showed that the Egfp-HA9801 cells grown at 37°C for 36 h displayed greater green fluorescence signals than the cells grown at 28°C for 36 h and 37°C for 24 h. The fluorescence in the tissue cryosections of Egfp-HA9801-injected mice was also stronger than that of the HA9801 group. Together, these results indicate that the egfp and spc(r) insertions into the Egfp-HA9801 recombinant did not significantly change the virulence when compared with HA980, and this EGFP labeled strain can be used for future S. suis 2 pathogenesis research.

Show MeSH
Related in: MedlinePlus