Mycoplasma Serotypes Research Articles

Mycoplasmas of goats and sheep.

The goat is an important commodity in many areas of the world, where it is kept as a source of meat, milk, and fiber. Often described as the “poor man’s cow,” the goat can survive in areas where a cow cannot and, therefore, replaces the cow in importance for a large segment of the world’s population. Improved goat husbandry will help maximize human food supplies from marginal agricultural lands under restrictive climatologic circumstances. Among the important goat diseases, mycoplasmal infections result in significant losses on the African continent and in countries such as Greece, France, Inassigned an arbitrary serotype number, except for ureaplasmas, which will be referred to as Ureaplasma spp. under a single serotype. To retain continuity with previous reports, serotype numbers for unclassified mycoplasmas are retained as originally listed in other reports. Three strains, Mycoplasma spp. G, U, and V, were first isolated from the external ear canals of clinically normal goats in Australia but also occur in the ear canals of goats in the United States (strains GM274B or GM790A, GM623, and GM257A, respectively) and are probably new mycoplasma species. New evdia, Israel, Italy, Portugal, Spain, and the United States. idence suggests that one of these serotypes (identical Morbidity and mortality can reach 100%. Several imto Mycoplasma sp. G or the US serologic equivalents portant outbreaks have been described, including 1 [strains GM274B or GM790A]) are pathogenic for the that required the destruction of an entire herd of 700 goat. goats in the United States. The list of mycoplasma serotypes given herein repClinical mycoplasmosis often lacks pathognomonic resents only those that differ from the other serotypes characteristics, and symptoms can be shared by or can named in this report, with the exception of strain mimic other clinically significant infections. As a conAl 343, which could not be obtained for comparison. sequence, the diagnosis of an acute caprine mycoplasThus, some publications list isolates as “unclassified” ma1 infection can be easily misinterpreted, even by the or “unspeciated” mycoplasma of goat or sheep oriveterinarian. For example, in acute or peracute infecgin 14,67,91 These mycoplasmas, and undoubtedly many tions with Mycoplasma capricolum or caprine strains others, are not included here because they were not of M. mycoides ssp. mycoides, the animal can die suddenly without premonitory signs. In such cases, the histologic sented for picion of gorized as changes are slight, and cases that are preroutine diagnostic analysis without any susmycoplasmal infection can easily be catebeing of undetermined etiology. serologically compared rotypes listed herein. with all the caprine/ovine se-

Electrophoretic analysis of proteins from Mycoplasma capricolum and related serotypes using extracts from intact cells and from minicells containing cloned mycoplasma DNA.

The acidic proteins of six different mycoplasma serotypes causing bovine or caprine pleuropneumonia were compared by two-dimensional gel electrophoresis of extracts of 35S-labelled cells. The organisms investigated were Mycoplasma mycoides subsp. mycoides (PG1), M. mycoides subsp. mycoides (Y-goat), M. mycoides subsp. capri (PG3), M. capricolum (California kid), the unclassified bovine serogroup 7 of Leach (PG50) and the F38-like group (F38). The results suggested a close relationship between M. capricolum and F38 and a similarly close relationship between the different M. mycoides subspecies, whereas the two M. mycoides subspecies appeared to be quite distant from M. capricolum and F38. The representative strain of the bovine serogroup 7 of Leach was equally distant from F38, M. capricolum and the three strains of M. mycoides. Strikingly, all six mycoplasma strains apparently shared six proteins in the two-dimensional gels. In Escherichia coli minicells, DNA from strain PG50 cloned in the vector pBR325 gave rise to incorporation of radioactive label into proteins which were identified as mycoplasma proteins by two-dimensional electrophoresis and immunoprecipitation.

Additional Information on the Classification of Avian Mycoplasma Serotypes

During the past year, we published an article in Avian Diseases discussing the cross-reactions among avian serotypes D (Mycoplasma gallinaceum) and F (Mycoplasma gallopavonis) using direct immunofluorescence (IF) (1). In that report, we described the unusual IF reactions between three test cultures of serotype D and a fluorescein-conjugated antiserum obtained from a rabbit immunized with serotype F (serotype F conjugate). These three D strains, designated Str. 8, TC-7, O(R), and DD, were IF-positive only with our serotype F conjugate, which was made from strain SA as shown in Table 1.

A Classification of Laboratory Strains of Avian Mycoplasma Serotypes by Direct Immunofluorescence

Direct immunofluorescence (IF) was used to assess cross-reactivity among 41 laboratory strains representing 11 serotypes of avian mycoplasmas. All the strains could be separated into distinct serotypes except the C-O-D-P group, which showed cross-relatedness unlike that demonstrated by other serological methods. With two exceptions, serotype C strains consistently cross-reacted with only the serotype P conjugate; the serotype O strain cross-reacted with the P and C conjugates. The two serotype C exceptions were found to be misclassified, as was the serotype P strain. Direct IF was found to be a quick, reliable method for measuring serological relatedness.

Neotetrazolium Staining of Three Avian Mycoplasma Serotypes for the Microagglutination Test

Mycoplasma meleagridis, M. gallisepticum, and M. synoviae antigens were stained with neotetrazolium chloride for use in the microagglutination test. Only the stained antigens of M. gallisepticum were acceptable in color intensity and test readability.

Differentiation of porcine mycoplasma strains by means of a simple paper chromatography test

Filter paper chromatography was used for the differentiation of the porcine mycoplasma. The test was sensitive and specific for the identification of the porcine mycoplasma serotypes and the results were in agreement with the metabolic inhibition test. It is a useful test for the identification of Mycoplasma hyorhinis.

A Division by Micro-Complement Fixation Tests of Previously Reported Avian Mycoplasma Serotypes into Identification Groups

A microtechnique complement-fixation (CF) test was used to establish identification groups among the previously described avian mycoplasma serotypes. The three named pathogenic species, Mycoplasma gallisepticum (serotype A), Mycoplasma meleagridis (serotype H), and Mycoplasma synoviae (serotype S), were each separable into a well-defined antigenic group, as were serotypes F and L. Predominantly one-way cross-relatedness was demonstrable between the other two named species included in this study, Mycoplasma gallinarum (serotype B) and Mycoplasma iners (serotype G). Serotype E was indistinguishable from serotype G by CF test. Crossrelatedness was also demonstrated among serotypes C, D, 0, and P and among serotypes I, J, K, N, Q, and R.

Interaction of porcine mycoplasmas with fresh animal serum.

When fresh animal serum was dropped onto seeded mycoplasma agar plates, inhibition of growth frequently occurred. This effect was dependent on the mycoplasma serotype and on the animal species from which the fresh serum came. This activity of fresh animal serum was heat-labile and would not diffuse through the agar medium. Growth of all the porcine mycoplasma serotypes was inhibited by fresh sheep serum. M. hyorhinis, M. hyopneumoniae, B 3 and the P 45 strains were insensitive to fresh horse serum. The addition of fresh horse serum to specific M. hyorhinis rabbit antiserum-impregnated disks appeared to have a synergistic effect and the combination of M. hyorhinis antiserum-impregnated disk and fresh horse serum always inhibited the growth of M. hyorhinis strains.

Classification of Avian Mycoplasmas by Gel-Diffusion and Growth-Inhibition Tests

SUMMARY The antigenic relations of 19 serotypes of avian mycoplasma were studied by immunodiffusion and growth inhibition. M. gallisepticum and M. synoviae had no cross reactions with other serotypes. According to the degree of cross reaction by both tests mentioned and the sharing of identical antigenic components by the gel-diffusion test, the 19 serotypes were classified into 9 groups. A modified disc growth inhibition with standardized inocula was adopted. This uses small quantities of antisera and is easy to interpret.

Polyacrylamide gel identification of bacterial L-forms and Mycoplasma species of human origin.

Polyacrylamide gel electrophoretic patterns of acidified phenol extracts prepared from whole cells can be used for the identification of bacterial L-forms and Mycoplasma species of human origin. Ten human Mycoplasma serotypes and eight L-forms belonging to five different genera were studied. The gel patterns were sufficiently distinct and reproducible that it was possible not only to identify L-forms at the genus level (group with streptococci) and different Mycoplasma serotypes but also to differentiate between the two of them. The parentage of L-forms of Streptobacillus moniliformis L1, Listeria monocytogenes, Streptococcus MG, and Staphylococcus aureus Smith strain was established by relating their gel patterns directly to parent bacteria. It was found that an L-form designated S. moniliformis An (ATCC 14220) was actually an L-form of Proteus. In addition, it was shown electrophoretically that no relationship existed between the Streptococcus MG L-form and M. pneumoniae. The applicability of this method as a diagnostic and taxonomic tool for the differentiation of L-forms and mycoplasmas is discussed.

Polyacrylamide Gel Identification of Bacterial L-Forms and Mycoplasma Species of Human Origin

Polyacrylamide gel electrophoretic patterns of acidified phenol extracts prepared from whole cells can be used for the identification of bacterial L-forms and Mycoplasma species of human origin. Ten human Mycoplasma serotypes and eight L-forms belonging to five different genera were studied. The gel patterns were sufficiently distinct and reproducible that it was possible not only to identify L-forms at the genus level (group with streptococci) and different Mycoplasma serotypes but also to differentiate between the two of them. The parentage of L-forms of Streptobacillus moniliformis L1, Listeria monocytogenes, Streptococcus MG, and Staphylococcus aureus Smith strain was established by relating their gel patterns directly to parent bacteria. It was found that an L-form designated S. moniliformis An (ATCC 14220) was actually an L-form of Proteus. In addition, it was shown electrophoretically that no relationship existed between the Streptococcus MG L-form and M. pneumoniae. The applicability of this method as a diagnostic and taxonomic tool for the differentiation of L-forms and mycoplasmas is discussed.

A Suggested Reclassification of Avian Mycoplasma serotypes

SUMMARY Avian Mycoplasma serotypes were examined serologically by the metabolic inhibition test. These cultures, previously described as 18 serotypes, fit 10 serologic groups. M. synoviae was not included in the study and M. meleagridis was included only in biochemic tests. Serologic groupings confirmed concurrent studies in which a variety of biochemic tests were used. Group A (M. gallisepticum) was not inhibited by heterologous antiserum. Group B (M. gallinarum) and M. anatis were antigenically related. Groups C and O were found to be identical as were groups D and P. Group E (M. iners), group F and group L were not inhibited by heterologous antiserum. Groups I, J, K, N, Q, and R were found to share common antigens and biochemic tests gave no basis for differentiation. Discrepancies with previous findings are attributed to: inadequate range of biochemic and growth characteristic tests; serologic and biochemic test data of limited value because of mistaken assumptions of pure cultures at the outset; and inadequate sensitivity of some serologic tests used earlier.

In vitro activity of lincomycin and spectinomycin against serotypes of avian mycoplasma.

Twenty strains of avian mycoplasma, representing 12 serotypes, were tested in vitro for their susceptibility to the action of lincomycin and spectinomycin alone and in combination. They varied in their sensitivity pattern. The ranges of minimal inhibitory concentration were 1 to 20 mug/ml for lincomycin or spectinomycin alone and 0.5/1 to 3/6 mug/ml for the lincomycin and spectinomycin combination. The ranges of minimal lethal concentration were greater with either single antibiotic than with the antibiotic combination. The amount of each antibiotic required to achieve mycoplasmacidal action of the relatively resistant strains was less with the antibiotic combination than with the single antibiotics.

Proposal for classifying strain PG-24 and related canine mycoplasmas as Mycoplasma edwardii sp. n.

Mycoplasmas recovered recently from dogs were found unrelated to three classified canine Mycoplasma serotypes but were similar in biological and serological properties to a Mycoplasma strain (C21, PG-24) isolated 18 years earlier. It is proposed that strains with the characteristics described be designated Mycoplasma edwardii sp. n.

Previously Uncharacterized Mycoplasma Isolates from an Investigation of Canine Pneumonia

Mycoplasmas recovered from the respiratory tract and genitourinary system of dogs, with and without respiratory infection, have been characterized by biological and immunological methods. Some of the isolates were indentified as being similar to the three species of canine mycoplasmas described earlier under the designation Mycoplasma spumans, M. canis, and M. maculosum. Other mycoplasmas placed in three groups (A, C, and D) were found to be clearly distinct from the three classified species. Group A strains fermented glucose but not mannose and were serologically distinct from other canine mycoplasmas recovered in this study. These strains were subsequently found to be biologically and serologically related to a previously reported, but unclassified, canine mycoplasma. Group D strains differed in some biological properties but were serologically related. These were found to be nonfermenting mycoplasmas representing isolations from the throat and bladder of dogs. They were serologically distinct from other canine mycoplasmas and were apparently unrelated to other known mycoplasma serotypes. Group C mycoplasmas were recovered only from the lungs of dogs. Within the group, they differ in some immunological properties but appear to be serologically distinct from other canine strains. They can also be separated from other dog strains in their ability to ferment glucose and mannose. Group B strains were found to have biological properties similar to M. canis strains but seemed to be only partially related to this serotype when examined in several serological techniques. It is suggested that these strains might represent antigenic variants of M. canis.

In Vitro Activity of Lincomycin and Spectinomycin Against Serotypes of Avian Mycoplasma

Twenty strains of avian mycoplasma, representing 12 serotypes, were tested in vitro for their susceptibility to the action of lincomycin and spectinomycin alone and in combination. They varied in their sensitivity pattern. The ranges of minimal inhibitory concentration were 1 to 20 mug/ml for lincomycin or spectinomycin alone and 0.5/1 to 3/6 mug/ml for the lincomycin and spectinomycin combination. The ranges of minimal lethal concentration were greater with either single antibiotic than with the antibiotic combination. The amount of each antibiotic required to achieve mycoplasmacidal action of the relatively resistant strains was less with the antibiotic combination than with the single antibiotics.

Utilization of neuraminic acid receptors by mycoplasmas.

Erythrocytes and H-HeLa cells were treated with neuraminidase and then compared with untreated cells for their ability to adsorb to mycoplasma colonies or be agglutinated by suspensions of the mycoplasmas. Of the 17 mycoplasma serotypes examined, only 4 were found to use neuraminic acid receptors; these were Mycoplasma pneumoniae, M. gallisepticum, M. synoviae, and mycoplasma WR1. Not all strains of a serotype behaved alike. Thus, removal of receptors on erythrocytes for one strain of M. gallisepticum required at least 100 times the concentration of neuraminidase needed to remove them for another strain. The mechanism of attachment of erythrocytes to mycoplasma colonies does not appear to be the same as that for attachment to mycoplasmas in suspension.

Immunofluorescence identification of Mycoplasma on agar by use of incident illumination.

The fluorescent-antibody method has been employed for the rapid identification of Mycoplasma colonies growing on agar plates. The method was found to be effective for detection of mixtures of Mycoplasma serotypes growing on primary isolation plates. The technique also helped to define the presence of mycoplasmas which did not produce typical colonies. It was also possible to identify Mycoplasma colonies overgrown by bacterial or fungal contaminants. Conjugates directed against 10 distinct Mycoplasma serotypes have been successfully employed in this system. One of the serotypes is a human oral isolate which has not been previously characterized.

Serum Fraction Requirements of Mycoplasma gallisepticum

Yoder and Hofstad (2) found that certain Mycoplasma serotypes survived eight passages in serum-free broth whereas an unclassified isolate persisted in serum-free media. Mycoplasma gallisepticum, however, was found to require serum for growth. Their report recalls an experience with an unsatisfactory serum fraction lot in this laboratory that justifies report since it represents a potential hazard in research and diagnostic procedure. One particular lot of PPLO serum fraction (Difco) obtained from an intermediary dealer failed to support growth of stock cultures of M. gallisepticum. Meanwhile, 3 unidentified Mycoplasma isolates grew readily in broth medium containing the offending lot. These three isolates did not produce sinusitis, other signs of disease, or M. gallisepticum agglutinins in turkeys or chickens when inoculated by appropriate respiratory routes. Further, unlike M. gallisepticum, they did not agglutinate chicken red blood cells.

Airsacculitis in Turkeys II. Use of Tylosin in the Control of the Egg Transmission of Mycoplasma spp. Other Than Mycoplasma gallisepticum in Turkeys

Airsacculitis in turkeys is one of the important causes of condemnations at the time the birds are inspected for wholesomeness. Field investigations and laboratory studies indicated that Mycoplasma gallisepticum and other Mycoplasma serotypes were associated with a high percentage of the turkeys showing airsacculitis at processing. It is well known that M. gallisepticum produces a severe airsacculitis in turkeys. Recent studies by Dierks (1) indicate that other Mycoplasma are also capable of causing airsacculitis.