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Phylogenetic analysis of the mycoplasma isolated from the Javanese macaque (M. fascicularis)
Abstract
The nucleotide sequence of the fragment of 16S-aRNA of mycoplasma was determined. The fragment was identified using the PCR method in the urogenital scrape of the Javanese macaque (M. fascicularis). The sequenced fragment of mycoplasma of M. fascicularis was compared to well-known sequences of mycoplasma of mammals. The results of our comparative and phylogenetic analyses of the sequenced DNA fragment revealed that the mycoplasma belonged to the M. primatum species and fell within the same cluster as M. hominis. The mycoplasma M. primatum was for the first time observed in the monkeys M. fascicularis. The pathogenicity of the mycoplasma species with respect to monkeys is being studied.
Mycoplasmas are primitive bacteria classified in the genus Mycoplasma (family Mycoplasmataceae), which can be cultured from healthy individuals encompassing a wide variety of vertebrate species. Most of the strains isolated from nonhuman primates, both Old World and New World, are ones that are considered human in origin. Disease attributed to Mycoplasma infection has been associated with polyarthritis in great apes and Old World monkeys. Antibiotic therapy has been used to treat these cases of infection.
Some strains of mycoplasmas and ureaplasmas (family Mycoplasmataceae) are associated with nongonococcal urethritis (NGU) or other genitourinary infections. We have developed a rapid and reliable
method of identifying the presence and prevalence of mycoplasmas and ureaplasmas in men with NGU. This method is based on
the amplification of a part of the 16S rRNA gene by PCR and phylogenetic analysis. A portion of the 16S rRNA gene from 15
prototype strains was amplified with a set of common primers, and their nucleotides were sequenced. The nucleotide sequence
of the V4 and V5 regions was analyzed by the neighbor-joining method. The 15 prototype strains were grouped into three distinct
clusters, allowing us to clearly segregate the strains into distinct lineages. To determine the prevalence of these pathogens
among patients with NGU, this protocol was tested with 148 urine samples. Amplifications were observed for 42 samples, and
their nucleotide sequences were analyzed along with those of the 15 prototype strains. The phylogenetic tree thus constructed
indicated that 15 of the 42 formed a cluster with Mycoplasma genitalium. Among the remaining specimens, 2 formed a cluster with Mycoplasma hominis, 19 with Ureaplasma urealyticum, and 5 with Ureaplasma parvum; the remaining sample contained both M. genitalium and U. urealyticum. This phylogeny-based identification of mycoplasmas and ureaplasmas provides not only a powerful tool for rapid diagnosis
but also the basis for etiological studies of these pathogens.
The mycoplasmal flora of the urinary tract of 19 men and 21 women was investigated at autopsy. Mycoplasmas were isolated from the urethra of 9 bodies. One of the isolates was identified biochemically and serologically as M. primatum. This is the second isolation from man of this simian mycoplasma species. Three of the remaining isolates from the urethra were identified as M. hominis, one as M. fermentans and four as U. urealyticum. U. urealyticum was isolated twice from the bladder urine. In one case, a strain of U. urealyticum was isolated from the cortex of a kidney with histologically demonstrable non-specific inflammation.
Nine adult female chimpanzees, five adult female orangutans, and three young female gorillas were cultured for mycoplasmas. All animals were found to harbor at least one species of mycoplasma and some as many as four. On the basis of physiological and serological characteristics mycoplasmas closely related or identical with M. hominis, M. salivarium, and M. orale types 1 and 2 were identified. The M. hominis strains which were restricted to the chimpanzee comprised two serologic subgroups but were physiologically homogeneous. The M. salivarium strains were physiologically and morphologically heterogeneous, with both lipolytic and non-lipolytic strains being isolated from the salivas of all three primate species. All strains of M. salivarium were closely related serologically. In addition to the above species, three unidentifiable types were encountered. Type 1 strains were isolated from the vaginas of two chimpanzees. The type 2 strain, which shared up to three antigenic components with various human mycoplasmas, was isolated from the oropharynx of a chimpanzee. The type 3 strains constituted the only mycoplasma species isolated from the orangutan vagina.
FOUR laboratories have collaborated to isolate and identify mycoplasmas from the pharyngeal and lower urogenital tissues of the African green monkey. Seventy mycoplasmas were isolated from fifty-two of 142 specimens obtained from seventy-one monkeys. Most of the simian mycoplasmas isolated were similar or identical to Mycoplasma species previously isolated only from man.
Mycoplasmas isolated from simian (Cercopithecus aethiops) tissues were shown to have biological, biochemical, and serological properties and electrophoretic cell protein patterns similar to strain Navel isolated from man 15 years ago. The simian and human Navel strains comprised a single serogroup, distinct from the established Mycoplasma and Acholeplasma species of the class Mollicutes. It is proposed that strains with the properties described be named Mycoplasma primatum.