Trimethylsilyl-sugar profiles of Streptococcus milleri and Streptococcus mitis

Seventy strains of 'viridans-group' streptococci were analysed gas chromatographically after preparation of trimethylsilyl ethers of their cellular sugars. The resulting profiles were evaluated as a possible aid to taxonomy. Glycerol, glucose, galactose, N-acetyl-glucosamine and N-acetylmuramic acid were found in all strains, in varying amounts. Rhamnose was the major neutral sugar in most strains, other than representatives of Streptococcus mitis , which invariably had ribose and usually anhydroribitol but no rhamnose. One strain of Strep, mitis possessed arabitol. Some strains of Strep. mitis and ' Strep. milleri ' were alone in containing N-acetylgalactosamine. A combination of N-acetylgalactosamine and rhamnose in the absence of ribose was diagnostic for strains of ' Strep. milleri '.     

Trimethylsilyl-sugar profiles of Streptococcus milleri and Streptococcus mitis

Seventy strains of 'viridans-group' streptococci were analysed gas chromatographically after preparation of trimethylsilyl ethers of their cellular sugars. The resulting profiles were evaluated as a possible aid to taxonomy. Glycerol, glucose, galactose, N-acetyl-glucosamine and N-acetylmuramic acid were found in all strains, in varying amounts. Rhamnose was the major neutral sugar in most strains, other than representatives of Streptococcus mitis, which invariably had ribose and usually anhydroribitol but no rhamnose. One strain of Strep. mitis possessed arabitol. Some strains of Strep. mitis and 'Strep. milleri' were alone in containing N-acetylgalactosamine. A combination of N-acetylgalactosamine and rhamnose in the absence of ribose was diagnostic for strains of 'Strep. milleri'.     

Inhibitory activity of Streptococcus mitis against oral bacteria

The antagonistic properties of three strains of Streptococcus mitis were investigated. They were found to inhibit a wide range of oral bacteria; Gram-positive and Gram-negative, facultative and anaerobic species being susceptible. The S. mitis strains were shown to be producing hydrogen peroxide, this being partially responsible for the aerobic inhibitory activity. A second inhibitory factor(s) was also produced, aerobically and anaerobically, although this could not be isolated. A limited characterization of this factor was undertaken using plate cultures.     

Role of Glycogen in Survival of Streptococcus mitis

Evidence has been obtained which indicates that the possession of glycogen by Streptococcus mitis favors its survival during starvation.     

A Virulent Babesia bovis Strain Failed to Infect White-Tailed Deer (Odocoileus virginianus)

Wildlife are an important component in the vector-host-pathogen triangle of livestock diseases, as they maintain biological vectors that transmit pathogens and can serve as reservoirs for such infectious pathogens. Babesia bovis is a tick-borne pathogen, vectored by cattle fever ticks, Rhipicephalus spp., that can cause up to 90% mortality in naive adult cattle. While cattle are the primary host for cattle fever ticks, wild and exotic ungulates, including white-tailed deer (WTD), are known to be viable alternative hosts. The presence of cattle fever tick populations resistant to acaricides raises concerns regarding the possibility of these alternative hosts introducing tick-borne babesial parasites into areas free of infection. Understanding the B. bovis reservoir competence of these alternative hosts is critical to mitigating the risk of introduction. In this study, we tested the hypothesis that WTD are susceptible to infection with a B. bovis strain lethal to cattle. Two groups of deer were inoculated intravenously with either B. bovis blood stabilate or a larval extract supernatant containing sporozoites from infected R. microplus larvae. The collective data demonstrated that WTD are neither a transient host nor reservoir of B. bovis. This conclusion is supported by the failure of B. bovis to establish an infection in deer regardless of inoculum. Although specific antibody was detected for a short period in the WTD, the PCR results were consistently negative at multiple time points throughout the experiment and blood from WTD that had been exposed to parasite, transferred into naïve recipient susceptible calves, failed to establish infection. In contrast, naïve steers inoculated intravenously with either B. bovis blood stabilate or the larval extract supernatant containing sporozoites rapidly succumbed to disease. These findings provide evidence that WTD are not an epidemiological component in the maintenance of B. bovis infectivity to livestock.     

Septicemia due to Streptococcus mitis in neutropenic patients with acute leukemia

Eight neutropenic patients with acute lymphocytic or nonlymphocytic leukemia had septicemia due to different strains of Streptococcus mitis (St. mitis), a microorganism not commonly recognized as a special pathogen in leukemic patients. Four of the patients had been treated with high-dose cytosine arabinoside as part of the cytostatic regimen, six had a central venous line and four patients had oral lesions prior to the infection. Selective gut decontamination consisted of co-trimoxazole/colistin in five patients and quinolones in three patients. The first three patients died, either due to interstitial pneumonia with the adult respiratory distress syndrome (ARDS), or due to infection-triggered disseminated intravascular coagulation despite prompt empiric antibiotic therapy including vancomycin. The other patients improved after empiric supplementation of penicillin G (30 Mega/day) to the antibiotic regimen. Beginning ARDS in two of these patients dramatically responded to high-dose steroids. We conclude that St. mitis is a major pathogen in neutropenic leukemic patients. Infection appears to occur independently of acute leukemic cell type, regimen of selective gut decontamination, venous access, visible oral lesions or treatment with high-dose cytosine arabinoside. The clinical course of our patients raises questions about the value of commonly recommended empiric antibiotic regimens, which were clearly ineffective to control infections with St. mitis in this patient group. Our data indicate that immediate antibiotic therapy with penicillin G is indicated and may be life-saving for suspected St. mitis infections in neutropenic leukemic patients.     

Metal ion inactivation and chelator stimulation of Streptococcus mitis arginine aminopeptidase

Activation of Streptococcus mitis (ATTC 9811) arginine aminopeptidase resulted in removal of the metal(s) from the enzyme molecule, and the action of the heavy metal ion in the inactivation process was shown to involve formation of a chelate complex between the enzyme molecule and metal or oxidation of functional group(s) on the enzyme surface. The enzyme also underwent activation by bovine serum albumin, amino acids, phosphate, and citric acid, which are probable physiological chelators.     

Commensal Streptococcus mitis is a unique vector for oral mucosal vaccination

The development of vaccine approaches that induce mucosal and systemic immune responses is critical for the effective prevention of several infections. Here, we report on the use of the abundant human oral commensal bacterium Streptococcus mitis as a delivery vehicle for mucosal immunization. Using homologous recombination we generated a stable rS. mitis expressing a Mycobacterium tuberculosis protein (Ag85b). Oral administration of rS. mitis in gnotobiotic piglets resulted in efficient oral colonization and production of oral and systemic anti-Ag85b specific IgA and IgG antibodies. These results support that the commensal S. mitis is potentially a useful vector for mucosal vaccination.     

Isoelectric focusing studies on the beta-fructofuranosidases and alpha-glucosidases of Streptococcus mitis.

Extracts of Streptococcus mitis ATCC 903 were analysed for beta-fructofuranosidase and alpha-glucosidase activities by isoelectric focusing in thin-layer polyacrylamide gels combined with zymogram procedures. Three bands of activity were visualized in the gels after incubation with sucrose (pI 4.05, 4.25 and 4.85) and three other bands after incubation with p-nitrophenyl alpha-D-glucopyranoside (pI 3.90, 4.45 and 4.65). The enzymes responsible for the reaction with sucrose were identified as beta-fructofuranosidases (EC 3.2.1.26) for the following reasons: identical enzyme bands were visualized in the gels after incubation with raffinose; no enzyme bands appeared in the gel after incubation with the alpha-glucosides maltose, turanose, trehalose and melezitose; and the soluble fraction hydrolysed sucrose to equimolar amounts of glucose and fructose.     

Evaluation of co-aggregation among Streptococcus mitis, Fusobacterium nucleatum and Porphyromonas gingivalis

AIMS: To develop a semi-quantitative method for evaluating co-aggregation reactions among three bacterial species, and to examine the influence of Fusobacterium nucleatum on the adherence of Porphyromonas gingivalis. METHODS AND RESULTS: The method involves coating hydroxyapatite (HAP) discs with streptococcal cells and treatment with radio-labelled bacterial cell suspensions. The sensitivity of the method was estimated by comparison with a turbidometric co-aggregation assay. Results from the two methods were in close agreement. Streptococcus mitis-coated HAP discs were immersed in a 3H-labelled Fus. nucleatum cell suspension and then a 14C-labelled P. gingivalis cell suspension. The discs were then pyrolysed to recover and quantify the released 3H and 14C radioactivity. The number of Fus. nucleatum cells on the discs increased with immersion time and this, in turn, resulted in elevated adherence of P. gingivalis. CONCLUSION: The data indicate that the method closely reflects co-aggregation characters, and that Fus. nucleatum has a positive effect on the adherence of P. gingivalis. SIGNIFICANCE AND IMPACT OF THE STUDY: The present method, which is designed to mimic the oral environment, should prove useful in the semi-quantitative evaluation of co-aggregation reactions.     

Identification of a secreted cholesterol-dependent cytolysin (mitilysin) from Streptococcus mitis

We have detected a cholesterol-dependent cytolysin, which we have named mitilysin, in a small number of Streptococcus mitis isolates. We have sequenced the mitilysin gene from seven isolates of S. mitis. Comparisons with the pneumococcal pneumolysin gene show 15 amino acid substitutions. S. mitis appear to release mitilysin extracellularly. Certain alleles of mitilysin are not recognized by a monoclonal antibody raised to the related toxin pneumolysin. Based on enzyme-linked immunosorbent assay and neutralization assay results, one isolate of S. mitis may produce a further hemolytic toxin in addition to mitilysin. As genetic exchange is known to occur between S. mitis and Streptococcus pneumoniae, this finding may have implications for the development of vaccines or therapies for pneumococcal disease that are based on pneumolysin.     

两株具有广谱抗菌性的缓征链球菌的安全性评价

目的 通过动物实验对分离于健康儿童口咽部的2株缓征链球菌菌株42、419进行安全性评价,为呼吸道益生菌的制备提供安全依据。 方法 对SPF级昆明种小鼠灌胃给予含有菌株42和419的菌液,观察实验动物是否出现死亡、急性毒性体征、体质量变化、采食量变化、血清淀粉样蛋白A变化和细菌移位生长情况,并对结果进行统计学分析。 结果 给药后,未发现实验小鼠死亡及急性毒性体征,未观察到细菌移位生长情况,给予菌株42后雌性和雄性剂量组与对照组间差异没有统计学意义(F=0.697 0,P=0.567 0;F=1.289 0,P=0.312 0)。给予菌株42的1 d后,雌性小鼠低、中剂量组较其他组别体质量变化量小(F=4.402 0,P=0.019 0),雄性小鼠高、中、低剂量组体质量变化量小于对照组(t=-0.800 0,P=0.035 0;t=1.000 0,P=0.011 0;t=-0.800 0,P=0.035 0);给予菌株419的1 d后,雌性、雄性小鼠体质量变化量与对照组差异无统计学意义(F=2.024 0,P=0.151 0;F=2.507 0,P=0.096 0)。给予菌株42的7 d内,雌性小鼠中剂量组和低剂量组采食量大于对照组(t=0.171 4,P结论 菌株42、419在为期7 d的急性毒性动物实验中,未发现毒性表现,可用于呼吸道益生菌菌株的选择。     

Multiple Diverse Circoviruses Infect Farm Animals and Are Commonly Found in Human and Chimpanzee Feces

Circoviruses are known to infect birds and pigs and can cause a wide range of severe symptoms with significant economic impact. Using viral metagenomics, we identified circovirus-like DNA sequences and characterized 15 circular viral DNA genomes in stool samples from humans in Pakistan, Nigeria, Tunisia, and the United States and from wild chimpanzees. Distinct genomic features and phylogenetic analysis indicate that some viral genomes were part of a previously unrecognized genus in the Circoviridae family we tentatively named “Cyclovirus” whose genetic diversity is comparable to that of all the known species in the Circovirus genus. Circoviridae detection in the stools of U.S. adults was limited to porcine circoviruses which were also found in most U.S. pork products. To determine whether the divergent cycloviruses found in non-U.S. human stools were of dietary origin, we genetically compared them to the cycloviruses in muscle tissue samples of commonly eaten farm animals in Pakistan and Nigeria. Limited genetic overlap between cycloviruses in human stool samples and local cow, goat, sheep, camel, and chicken meat samples indicated that the majority of the 25 Cyclovirus species identified might be human viruses. We show that the genetic diversity of small circular DNA viral genomes in various mammals, including humans, is significantly larger than previously recognized, and frequent exposure through meat consumption and contact with animal or human feces provides ample opportunities for cyclovirus transmission. Determining the role of cycloviruses, found in 7 to 17% of non-U.S. human stools and 3 to 55% of non-U.S. meat samples tested, in both human and animal diseases is now facilitated by knowledge of their genomes.Animal viruses with small, circular, single-stranded DNA (ssDNA) genomes comprise the Circoviridae family and the Anellovirus genus, while viruses in the Geminiviridae and Nanoviridae families infect plants (3, 25, 34, 37, 40). The genomes of these small viruses without a lipid envelope replicate through a rolling-circle mechanism, possibly sharing a common origin with bacterial plasmids (6), and show high recombination and nucleotide substitution rates (7, 19).The Circoviridae family consists of the Circovirus genus whose member species are currently known to infect only birds and pigs, and the Gyrovirus genus, including a single species, Chicken anemia virus (CAV). Circoviruses infect several avian groups, including parrots, pigeons, gulls, anserids (ducks, geese, and swans), and numerous passerines (ravens, canaries, finches, and starlings) (12, 15, 16, 22, 26, 31, 35, 38, 39). Avian circoviruses have been associated with a variety of symptoms, including developmental abnormalities, lymphoid depletion, and immunosuppression (22, 26, 28, 35, 39). Mammalian circoviruses include only two closely related species, Porcine circovirus 1 and 2 (PCV1 and PCV2, respectively), infecting pigs (21). PCV2 has been associated with porcine circovirus-associated diseases, which can manifest as a systemic disease, respiratory disease complex, enteric disease, porcine dermatitis and nephropathy syndrome or as reproductive problems, causing great losses in the pork industry (1, 29, 32). Circovirus infections are thought to occur mainly through fecal-oral transmission (37).We describe here highly diverse, circovirus-like, circular DNA viral genomes discovered in human and chimpanzee stool samples, and we propose their inclusion in a new genus of the Circoviridae family that we tentatively name “Cyclovirus” pending review by the International Committee on Taxonomy of Viruses (ICTV). Cycloviruses were also found to be prevalent in the muscle tissue of farm animals, such as chickens, cows, sheep, goats, and camels. The cyclovirus species found in human stool samples and in animal meat samples showed limited genetic overlap, suggesting that most of the cycloviruses found in human stool samples are not from consumed animal meat. Rather, these cycloviruses in human stools might cause human enteric infections. The presence of cycloviruses in human stool samples and in farm animal tissue also suggests the potential for frequent cross-species exposure and zoonotic transmissions.     

Virulent aggregates of Streptococcus pyogenes are generated by homophilic protein-protein interactions

Many strains of the important human pathogen Streptococcus pyogenes form aggregates when grown in vitro in liquid medium. The present studies demonstrate that this property is crucial for the adherence, the resistance to phagocytosis and the virulence of S. pyogenes. A conserved sequence of 19 amino acid residues (designated AHP) was identified in surface proteins of common S. pyogenes serotypes. This sequence was found to promote bacterial aggregation through homophilic protein-protein interactions between AHP-containing surface proteins of neighbouring bacteria. A synthetic AHP peptide inhibited S. pyogenes aggregation, reduced the survival of S. pyogenes in human blood and attenuated its virulence in mice. In contrast, mutant bacteria devoid of surface proteins containing AHP-related sequences did not aggregate or adhere to epithelial cells. These bacteria are also rapidly killed in human blood and show reduced virulence in mice, underlining the pathogenic significance of the AHP sequence and S. pyogenes aggregation.