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Identification of streptococci to species level by sequencing the gene encoding the manganese-dependent superoxide dismutase.

We have used a PCR assay based on the use of degenerate primers in order to characterize an internal fragment (sodA(int)) representing approximately 85% of the genes encoding the manganese-dependent superoxide dismutase in various streptococcal type strains (S. acidominimus, S. agalactiae, S. alactolyticus, S. anginosus, S. bovis, S. constellatus, S. canis, S. cricetus, S. downei, S. dysgalactiae, S. equi subsp. equi, S. equi subsp. zooepidemicus, S. equinus, S. gordonii, S. iniae, S. intermedius, S. mitis, S. mutans, S. oralis, S. parasanguis, S. pneumoniae, S. porcinus, S. pyogenes, S. salivarius, S. sanguis, S. sobrinus, S. suis, S. thermophilus, and S. vestibularis). Phylogenetic analysis of these sodA(int) fragments yields an evolutionary tree having a topology similar to that of the tree constructed with the 16S rRNA sequences. We have shown that clinical isolates could be identified by determining the positions of their sodA(int) fragments on the phylogenetic tree of the sodA(int) fragments of the type species. We propose this method for the characterization of strains that cannot be assigned to a species on the basis of their conventional phenotypic reactions.

Base Sequence↗

The role of international transport of equine semen on disease transmission.

Despite the numerous benefits of having the capability to transport semen internationally, there are serious potential ramifications if that semen is contaminated with a communicable disease. Bacteria: Many commensal bacteria colonize the exterior of the stallion penis and are not regarded as pathogenic. They may be cultured from an ejaculate. Alterations of the normal bacterial flora on the exterior genitalia may cause the growth of opportunistic bacteria such as Klebsiella pneumonia, Pseudomonas aeruginosa, Streptococcus zooepidemicus, which, if inseminated, may cause infertility in susceptible mares. Contagious equine metritis (CEM), a highly transmissible, true venereal disease of horses, is caused by the gram-negative coccobacillis, Taylorella equigenitalis. Even with the use of rigorous testing protocols, the current techniques used may not ensure accuracy of results. Viruses: Equine coital exanthema (equine herpes virus type 3; EHV-3) is a highly contagious virus that causes painful lesions on the stallion's penis and mare's vulva. Although it is primarily transmitted through coitus, infected fomites have also been implicated in its spread. Therefore, it is possible that the virus can potentially be transmitted to the ejaculate through penile contact with an artificial vagina or sleeve. Equine arteritis virus appears to be becoming more prevalent in recent years. The most common method of transmission is through respiratory disease, but the organism can also be shed in the semen of asymptomatic stallions. Equine infectious anemia virus has also been found to be present in the semen of an infected stallion, although no evidence exists at this time that there is venereal transmission of this disease. Protozoa: Dourine, caused by Trympanosoma equiperidum, is a venereal disease found only in Africa, South and Central America and the Middle East. Serological testing using complement fixation is recommended for diagnosis. Piroplasmosis, a disease caused by Babesia equi or by a less severe strain, Babesia caballi, has received a great deal of attention in recent years due to the increased transfer of horses between countries. It is considered to be enzootic in many areas of the southern US, and is found throughout the world. The protozoal agent is most often spread by ticks, but mechanical transmission has also been documented; therefore, there is concern for venereal transmission if blood from an infected horse contaminates the semen.

Animals↗

Specific binding sites for monomeric and aggregated beta 2-microglobulin on surface of groups A, B, C, and G streptococci.

Human beta 2-microglobulin (beta 2-m) was isolated from urine samples of patients with tubular dysfunctions and aggregated with glutaraldehyde. Four aggregates with molecular weights of 800,000, 480,000, 260,000, and 60,000 were separated by filtration on Sephacryl S-300. The aggregates and monomeric beta 2-m (11,800 MW) were subsequently labeled with 125I and tested for binding to streptococci. Group A streptococci bound only aggregated beta 2-m with a mean binding of 44.5%. Most of the group G streptococci, on the other hand, bound only monomeric beta 2-m with a mean binding of 58%. Among group B streptococci the serotypes with protein antigens interacted mainly with monomeric beta 2-m and those without protein antigens preferentially with aggregated beta 2-m. Nontypable group B streptococcal serotypes did not bind monomeric or aggregated beta 2-m. Of the streptococci belonging to group C, S. equisimilis reacted with monomeric beta 2-m and S. dysgalactiae with aggregated beta 2-m. S. equi did not interact with monomeric beta 2-m or aggregated beta 2-m. Bindings of monomeric beta 2-m and aggregated beta 2-m were saturable and could be inhibited by the respective unlabeled forms of beta 2-m. Fibrinogen, fibronectin, alpha 2-macroglobulin, haptoglobin, or immunoglobulin G did not inhibit the binding of either form of beta 2-m. The binding sites for monomeric beta 2-m were more susceptible to trypsin than those for aggregated beta 2-m. Treatment of streptococci with pronase destroyed their binding activities for monomeric and aggregated beta 2-m. Both monomeric beta 2-m and aggregated beta 2-m binding sites were sensitive to heat. The Scatchard plots of monomeric beta 2-m and aggregated beta 2-m were linear with Kd of 1.29 X 10(-9) M and 1.9 X 10(-9) M respectively. The number of binding sites per bacterium were estimated to be 81,000 for monomeric beta 2-m and 1,210 for aggregated beta 2-m.

Binding Sites↗