Isolation Of Mycoplasmas Research Articles

Association of Raillietia caprae with the presence of Mycoplasmas in the external ear canal of goats

We did a descriptive study to determine whether the presence in the external ear canal of the Raillietia caprae mites and Mycoplasmas were associated. For that we sampled 360 goats slaughtered at abattoirs in the summer to identify those infested with the mite. We found only 20 infested, so used all of those plus another 47 uninfested goats selected systematically from the population negative for the isolation of Mycoplasmas. These goats came from the regions of Queretaro, Guanajuato, Sinaloa and Estado de México. Sterile swabs were taken from each ear canal of the carcass after removal of the pinna for microscopic observation of the mites and for the isolation of Mycoplasmas in both study groups. The swab samples were inoculated in Friis media for the isolation of Mycoplasmas; then, the isolates were biochemically characterized and identified serologically. We recovered isolates from the earwax of only nine of the 47 control goats, but from the earwax of 11 of the 20 infested goats; another four infested goats had Mycoplasma isolated from the mites but not from the earwax. Mycoplasma cottewii and Mycoplasma yeatsii were the only Mycoplasmas isolated from the uninfested goats, and also were the predominant (29 of 34) isolates from the infested goats and/or from the mites.

Mycoplasmas Isolated from Stone Curlews (Burhinus oedicnemus) Used in Falconry in the United Arab Emirates

The aim of this study was to evaluate the risk of transmission of Mycoplasma spp. from quarry to hunting falcons in the Middle East. Groups of 17 houbara bustards (Chlamydotis undulata) and 29 stone curlews (Burhinus oedicnemus) kept at three different private collections in Dubai were evaluated for the presence of Mycoplasma. Additionally, 10 falcons used for hunting were investigated for comparison. The falcons showed no clinical signs and were examined within the scope of a routine health check. From all birds, conjunctival and choanal swabs were taken and analyzed via polymerase chain reaction and culture. Although mycoplasmas were not recovered from choanal and conjunctival swabs taken from the houbara bustards, Mycoplasma gypis and M. falconis were isolated from the majority (28/29; 97%) of the stone curlews from choanal and conjunctival swabs. Most of the birds had no associated pathologic findings. Mycoplasma falconis was also detected in samples collected from 2 of the 10 falcons, and M. buteonis was isolated from the majority of falcons (6/10 falcons) from choanal (n = 5) and conjunctival (n = 1) swabs. Mycoplasma gypis could also be isolated from tissue samples (liver, oviduct, syrinx) of one dead stone curlew. This study presents the first isolation of mycoplasmas from stone curlews.

Raman spectroscopic typing reveals the presence of carotenoids in Mycoplasma pneumoniae

Raman spectroscopy has previously been demonstrated to be a highly useful methodology for the identification and/or typing of micro-organisms. In this study, we set out to evaluate whether this technology could also be applied as a tool to discriminate between isolates of Mycoplasma pneumoniae, which is generally considered to be a genetically highly uniform species. In this evaluation, a total of 104 strains of M. pneumoniae were analysed, including two reference strains (strains M129 and FH), and 102 clinical isolates, which were isolated between 1973 and 2005 and originated from various countries. By Raman spectral analysis (Raman typing) of this strain collection, we were able to reproducibly distinguish six different clusters of strains. An unequivocal correlation between Raman typing and P1 genotyping, which is based on sequence differences in the P1 (or MPN141) gene of M. pneumoniae, was not observed. In the two major Raman clusters that we identified (clusters 3 and 6, which together harboured 81 % of the strains), the different P1 subtypes were similarly distributed, and approximately 76 % isolates were of subtype 1, approximately 20 % of subtype 2 and approximately 5 % of variant 2a. Nevertheless, a relatively high prevalence of P1 subtype 2 strains was found in clusters 2 and 5 (100 %), as well as in cluster 1 (75 %) and cluster 4 (71 %); these clusters, however, harboured a small number of strains. Only two of the strains (2 %) could not be typed correctly. Interestingly, analysis of the Raman spectra revealed the presence of carotenoids in M. pneumoniae. This finding is in line with the identification of M. pneumoniae genes that have similarity with genes involved in a biochemical pathway leading to carotenoid synthesis, i.e. the 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway. Therefore, we hypothesize that M. pneumoniae hosts an MEP-like pathway for carotenoid synthesis. We conclude that Raman spectroscopy is a convenient tool for discriminating between M. pneumoniae strains, and that it presents a promising supplement to the current methods for typing of this bacterium.

Antimicrobial Susceptibilities ofUreaplasma urealyticumandMycoplasma hominisin Pregnant Women

Background Ureaplasma urealyticum and Mycoplasma hominis are associated with an increased risk of pregnancy complications, such as preterm birth and premature membrane rupture. The purpose of this study was to determine the isolation rates and antimicrobial susceptibilities of genital mycoplasma in a sample of pregnant women from Jinju, Korea. Methods Vaginal swabs were obtained from 258 pregnant women between 2004 and 2008 and tested for the presence of U. urealyticum and M. hominis at Gyeongsang National University Hospital. The identification and antimicrobial susceptibilities of U. urealyticum and M. hominis were determined with a commercially available kit, the Mycoplasma IST2 Kit (bioMé-rieux, Marcy-l'Etoile, France), and evaluated according to standards set by the Clinical and Laboratory Standards Institute (CLSI). Results U. urealyticum only was detected in 105 specimens (38.6%), while M. hominis only was detected only in 2 specimens (1.8%). Seven specimens (6.7%) were positive both for U. urealyticum and M. hominis. Susceptibilities of U. urealyticum to azithromycin, erythromycin, clarithromycin, and doxycycline were 75.2%, 82.9%, 88.6%, and 88.6%, respectively, while almost all of the isolates were susceptible to josamycin (99.0%) and pristinamycin (100%). The susceptibility of U. urealyticum to ofloxacin and ciprofloxacin was 56.2% and 15.2%, respectively. Conclusion The rate of isolation of genital mycoplasma in pregnant women was 44.2% in Jinju; most of the mycoplasma were U. urealyticum. U. urealyticum and M. hominis were highly resistant to quinolones, but susceptible to josamycin. Therefore, empirical treatment without prior identification and determination of the antimicrobial susceptibility of genital mycoplasma will fail in many cases.

The Role of Mycoplasmas in a Conservation Project of the Lesser Kestrel (Falco naumanni)

The lesser kestrel (Falco naumanni) is one of the most endangered bird species in Europe, and a captive breeding and reintroduction project was established. A breeding project is vulnerable to pathogens, e.g., mycoplasmas, reducing the reproductive success and carrying the risk to release pathogens with the birds to the wild. Therefore, 18 infertile eggs and 43 dead in shell embryos of the breeding project, as well as 27 nestlings and 34 adult birds of the captive and three different free-ranging populations were investigated for the occurrence of mycoplasmas by culture and a Mycoplasma genus-specific polymerase chain reaction. All eggs, embryos, and hand-reared nestlings from the captive group were negative. In contrast, all parent-reared nestlings and 88% of the adults were positive. Mycoplasma falconis and unidentifiable mycoplasmas were detected in all groups. Mycoplasma buteonis was found in the captive and only in two of the three free-ranging populations. Sequencing the 16S rRNA gene of six randomly selected unidentified isolates showed that five isolates were similar and most likely had been found previously in a falcon from Germany. The remaining isolate demonstrated a very high homology to unidentified Mycoplasma isolates obtained previously from semen samples of raptors. The results suggest that these isolates might represent two new species. Mycoplasmas seem not to play a major role as pathogens in the breeding project, and there is no evidence that releasing birds poses a risk to the free-ranging population with regard to mycoplasmas. The study seems to be the first to describe the occurrence and role of mycoplasmas in the lesser kestrel.

Occurrence of mycoplasmas in semen samples of birds of prey

Mycoplasmas are well-known pathogens in a variety of animals. In poultry it is known that some species can be transmitted by semen and infect the uterus of females. As the prevalence of mycoplasmas in birds of prey is very high and artificial insemination is a commonly used technique for reproduction, the possibility of transmission Mycoplasma spp. by contaminated semen in birds of prey was investigated. Isolation of mycoplasmas was possible in five out of 32 (15.6%) semen samples of different bird of prey species. Two additional semen samples were positive for mycoplasma DNA using a Mycoplasma-genus-specific polymerase chain reaction. The isolation of mycoplasmas from a testicular sample indicates the testis as the possible source of contamination. Sequencing of large parts (>90%) of the 16S rRNA gene of the isolated mycoplasmas suggests that all isolates belong to the same species. Alignment of the sequenced products with the 16S rRNA gene of Mycoplasma species in GenBank demonstrated a similarity of 97% to Mycoplasma verecundum, but serological testing by immunobinding assay failed to identify it as such. It is recommended that the semen of donor birds of prey is examined for mycoplasmas before its use in artificial insemination.

Activity of moxifloxacin against the urogenital mycoplasmas Ureaplasma spp., Mycoplasma hominis and Mycoplasma genitalium and Chlamydia trachomatis

The activity of moxifloxacin was compared with that of other antimicrobial agents against 54 strains of Ureaplasma spp., 54 strains of Mycoplasma hominis, 14 strains of Mycoplasma genitalium, and 44 strains of Chlamydia trachomatis. Moxifloxacin inhibited 90% of all isolates at a concentration </=1 mg/L, being the most active compound against C. trachomatis and sharing the highest activity with garenoxacin and gemifloxacin against mycoplasmas. Moxifloxacin killed the 30 mycoplasma isolates tested at a concentration </=1 mg/L, except those resistant to fluoroquinolone. Thus, moxifloxacin has attracted interest as a potential therapy for mycoplasmal or chlamydial urogenital infections.

Comparative In Vitro Activities of the Investigational Fluoroquinolone DC-159a and Other Antimicrobial Agents against Human Mycoplasmas and Ureaplasmas

The in vitro susceptibilities of 151 unique clinical isolates of Mycoplasma pneumoniae, Mycoplasma hominis, Mycoplasma fermentans, Mycoplasma genitalium, and Ureaplasma species to DC-159a, an investigational fluoroquinolone, in comparison with those to other agents were determined. Macrolides were the most active agents against M. pneumoniae and M. genitalium, whereas clindamycin was most active against M. hominis. DC-159a MICs were <or=0.5 microg/ml for all Mycoplasma species and <or=4 microg/ml for ureaplasmas. DC-159a was the most active fluoroquinolone tested against M. pneumoniae and M. fermentans, and it was second to moxifloxacin against the other species. It was bactericidal against 10 M. pneumoniae isolates and demonstrated killing of >or=99.9% of the inoculum at 24 h for 2 isolates. The excellent in vitro activity of DC-159a demonstrates its potential for use in the treatment of infections due to mycoplasmas and ureaplasmas.

Species-Specific Polymerase Chain Reactions for the Detection of Mycoplasma buteonis, Mycoplasma flconis, Mycoplasma gypis, and Mycoplasma corogypsi in Captive Birds of Prey

Mycoplasmas are pathogens of different avian species, but the role of Mycoplasma in raptors is not yet completely determined. As Mycoplasma isolation and identification present several difficulties, species-specific polymerase chain reactions (PCRs) for the detection of mycoplasmas found in birds of prey (Mycoplasma buteonis, Mycoplasma corogypsi, Mycoplasma falconis, and Mycoplasma gypis) were established. The specificity of the PCR methods were investigated using known avian Mycoplasma reference strains and isolates as well as related bacteria and was found to be specific. Amplificons obtained with these PCRs from field samples showed no false-positive results in restriction enzyme analysis and sequencing. The sensitivities of the different PCR assays varied between 50 fg and 1 pg DNA. Twenty-five tracheal swabs from healthy captive birds of prey were investigated by culture and immunobinding assay as comparison to the PCRs. Mycoplasmal DNA was detected in 88% of the samples, with negative results only from vultures. Mycoplasma falconis and M. buteonis were regularly found in falcons, and M. gypis was found in a common buzzard. Mycoplasma corogypsi was not demonstrated. Several isolates could not be differentiated using an immunobinding assay as well as the described PCR methods.

Real-time polymerase chain reaction assays for the detection of members of the Mycoplasma mycoides cluster

AIM: To develop real-time PCR assays for the detection and differentiation of members of the Mycoplasma mycoides cluster. METHODS: Five real-time PCR assays were designed to allow differentiation of members of the M. mycoides cluster: an assay for detection of the M. mycoides subspecies, viz M. mycoides subsp mycoides large colony (MmmLC), M. mycoides subsp capri (Mmc), and M. mycoides subsp mycoides small colony (MmmSC); one for the detection of the M. capricolum subspecies, viz M. capricolum subsp capricolum (Mcc), M. capricolum subsp capripneumoniae (Mccp), and Mycoplasma sp bovine group 7 (BG7); and three for the specifi c detection of MmmSC, Mccp, and BG7. A panel of 74 Mycoplasma isolates from various geographical origins and a panel of 21 other bacterial isolates were used to evaluate the sensitivity and specificity of the assays. RESULTS: The assays displayed 100% analytical sensitivity in detecting all target Mycoplasma isolates. The analytical detection limit for the assays to detect the M. mycoides subspecies, M. capricolum subspecies, and MmmSC was determined to be 100 fg of genomic DNA, while the Mccp and BG7 assays had a detection limit of 100 fg and 10 fg of genomic DNA, respectively. The M. mycoides subspecies assay had a detection limit of 103 (SD 102) cfu/ml milk, 104 (SD 104) cfu per swab, and 103 (SD 103) cfu/g lung in inoculated samples. The assays displayed 100% specificity when applied to non-target bacterial isolates and to 110 culture-negative milk samples. CONCLUSIONS: The assays were highly sensitive and specific, and provide accurate detection and differentiation of the members of the M. mycoides cluster.

Isolation and Characterization of Unusual Mycoplasma spp. from Captive Eurasian Griffon (Gyps fulvus) in Sicily

Mycoplasmas have been isolated from birds of prey during clinical examinations, but their significance to the health of raptors is unclear. We report the isolation and characterization of four mycoplasmas found in the upper respiratory tract of four sick Eurasian Griffon (Gyps fulvus) that were housed in a Sicilian rehabilitation center at Ficuzza, near Palermo in Sicily, before reintroduction into the wild. These included Mycoplasma gallinarum, an unidentified mycoplasma highly similar to Mycoplasma glycophilum, and two unidentified mycoplasmas with similarities to Mycoplasma falconis and Mycoplasma gateae.

In vitro antimycoplasmal activity of six Jordanian medicinal plants against three Mycoplasma species

The in vitro effect of six Jordanian traditional medicine plant methanolic extracts were tested against 32 isolates of Mycoplasma species; Mycoplasma mycoides subsp. mycoides LC (6), Mycoplasma capricolum subsp. capricolum (8) and M. putrefaciens (18), all isolated from either nasal swabs or milk, from sheep and goats in different regions in Jordan. All Mycoplasma species showed susceptibility to Artemisia herba-alba and Artemisia arborescens with MIC ranges from 3.125-12.5 mg/ml. Allium sativum and Punica grantum showed limited activity against some Mycoplasma isolates. Olea europea and Citrullus colocynthis showed no in vitro activity against any of the Mycoplasma species tested. Artemisia herba-alba and Artemisia arborescens may therefore be useful for the treatment of mycoplasma infections.

Identification and differentiation of canine Mycoplasma isolates by 16S–23S rDNA PCR-RFLP

Conventional serological methods for the identification of canine mycoplasma isolates depend on the availability of a panel of species-specific diagnostic antisera and are not always reliable in terms of specificity. To enable simultaneous identification of field isolates, PCR-RFLP analysis of the 16S–23S rRNA intergenic spacer region was used to characterize the type strains of the 12 currently described canine mycoplasmas of the Genus Mycoplasma which represent the “classic” non-hemotropic species. The use of 16S–23S rDNA PCR in the first step of this analysis revealed specific size differences of amplicons which allowed to classify these 12 canine Mycoplasma species into three groups. Depending on the length of the amplicon, subsequent RFLP analysis of PCR products using two restriction endonucleases in a single digest (ApoI/DdeI or TaqI/VspI) generated unique banding patterns. For further evaluation of the 16S–23S rDNA PCR-RFLP assay system as identification and differentiation tool, a total of 262 field isolates collected from the canine genital tract were tested. PCR-RFLP results for 251 field isolates correlated with traditional serological test results. The remaining 11 isolates had an RFLP pattern distinct from the type strains included in this study and were identified by 16S rDNA sequencing as closely related to M. sp. HRC689. The PCR-RFLP assay established in this study enabled a rapid, accurate and easily performed identification and differentiation of all 12 currently described non-hemotropic canine Mycoplasma species.

Prevalence of mycoplasmas in external ear canal of goats: Influence of the sanitary status of the herd

A study was carried out in 15 French goat herds to assess the presence of mycoplasmas in the external ear canal of goats and to estimate the association between the ear isolates (highly pathogenic or non-pathogenic) and the status of the herds with respect to the mycoplasma infections. In each herd, 20 clinically normal goats were investigated once and mycoplasma isolates were identified by MF-dot tests. Influence of herd status on the proportion of animals carrying highly pathogenic species of mycoplasmas was evaluated by calculating the Odds Ratios. The present study gave first of all an estimate of the relative importance of mycoplasma carriage in healthy French dairy goat herds. The species isolated were MmmLC, Mcc, Ma, Mp (considered as highly pathogenic) and My and Mc (non-pathogenic species). Carriage often involve several strains and/or species in the same ear canal. The highly pathogenic species were significantly more frequent in the herds with prior history of mycoplasma infection. This asymptomatic carrier state makes the sanitary status of a herd ambiguous and represents a risk of subsequent propagation of the etiological agents, during close physical contact or by intervention of vectors (mites).

Effect of danofloxacin (Advocin A180) on goats affected with contagious caprine pleuropneumonia

The efficacy of danofloxacin (Advocin A180) was evaluated for the treatment of contagious caprine pleuropneumonia (CCPP) caused by Mycoplasma capricolum subsp. capripneumoniae. Ten healthy Angora goats, confirmed free of CCPP, were exposed to clinically affected animals from a natural outbreak in Thrace, Turkey. After 14 days exposure, 8 goats showed pyrexia ( > or = 41 degrees C). Shortly after, the Angora goats were divided randomly into two groups. Five of these were injected with danofloxacin (6 mg/kg subcutaneously), which was repeated after 48 h; the five remaining animals received saline. Goats were monitored clinically and blood samples were collected for serology. Animals with severe disease were withdrawn from the trial. Goats completing the study were euthanized at day 42. Lung tissue and bronchial fluid were collected for mycoplasma isolation. All danofloxacin-treated goats showed resolution of clinical disease by the end of the trial. Two saline-treated goats failed to complete the study owing to CCPP. Danofloxacin-treated goats showed fewer lung lesions and had significantly lower combined clinical scores than saline controls (p < 0.001). Danofloxacin was found to be highly effective in the treatment of CCPP in goats.

An analysis of Detections and Drug susceptibility of Chlamydia Trachomatis and Mycoplasma in Infertile Women

目的 了解不孕症患者沙眼衣原体(CT)、解脲支原体(UU)和人型支原体(MH)感染状况及支原体药物敏感性.方法对3856例不孕症患者(不孕组)进行CT、UU和MH检测,CT采用爱尔兰ELISA检测抗原,支原体培养和药敏试验采用法国梅里埃公司提供的支原体培养、鉴定、药物一体化试剂盒.结果 (1)不孕组衣原体和支原体总感染率为66.83%,显著高于已孕组的15.17%(P 0.001);不孕组CT和UU感染率分别为12.19%和44.84%,显著高于已孕组的3.37%和8.99%(P 0.001).(2)UU、MH、UU+MH对前三位敏感的抗生素分别是交沙霉素、强力霉素、阿齐霉素,耐药率最高的前三位抗生素分别是氧氟沙星、环丙沙星、四环素.结论对不孕症患者进行衣原体和支原体检测及药敏试验,可以针对性地进行抗感染治疗,是解除衣原体和支原体感染所致不孕症患者的痛苦,恢复其正常生育功能的有效措施。

Prevalence of contagious caprine pleuropneumonia in the Borana pastoral areas of Ethiopia

The study was conducted in selected districts of Borana pastoral areas of Ethiopia, namely Yabello, Dire, Moyale and Liban, during June–September 2004 to determine the status of contagious caprine pleuropneumonia (CCPP). The study includes a retrospective study, fieldwork, serology, an abattoir investigation and isolation of the causative Mycoplasma. Outbreaks of CCPP were reported from almost all regions of the country, especially from lowland areas, which are known goat-rearing regions, with the highest in 2002. The predominant clinical findings observed in affected flocks were coughing, nasal discharge, weakness, reluctance to move, abduction of elbow and laboured breathing. A total of 35 sheep and goats, of 183 examined, were sero-positive for MCCP F 38 antibodies using CFT. From the total of 217 goats examined in the abattoir for the presence of CCPP lesions, 21 (9.7%) showed pathological lesions, inflamed lungs with marbled appearance, fibrous pleuropneumonia, yellowish-coloured pleural fluid and swollen bronchial and mediastinal lymph nodes. The causative agent ( Mycoplasma capricolum sp. capripneumoniae) was isolated and identified from tissue samples and thoracic fluid treated bacteriologically. In conclusion, the study indicates that CCPP is becoming a very important goat disease in Borana pastoral areas.

Small ruminant mycoplasmoses with particular reference to the Middle East.

Abstract Mycoplasmas are responsible for many human and animal diseases and have a tremendous economic and health impact worldwide. They have a pronounced affinity for mucosal surfaces and consequently cause infections of the urogenital and respiratory tracts, but also of the mammary gland and serous membranes, particularly in animals. Mycoplasma infections of small ruminants are known to exist worldwide and cause significant economic impacts because of substantial reductions in milk yield, meat, skins and hair in affected animals; however, little is known of their occurrence in the Middle East. Many mycoplasmas can infect lungs of small ruminants and induce respiratory disease and these Mycoplasma capricolum subsp. capripneumoniae , M. mycoides subsp. capri [formerly M. mycoides subsp. mycoides LC (International Committee on Systematics of Prokaryotes Subcommittee on the Taxonomy of Mollicutes have proposed the merging of these two subspecies into the single subspecies: M. mycoides subsp. capri ; a decision is pending.)] and Mycoplasma capricolum subsp. capricolum (Mcc). M. agalactiae and M. putrefaciens can be a major cause of mastitis, arthritis and ocular disease. This review highlights the importance of pathogenic mycoplasmas in small ruminants, including M. putrefasciens which has never been described previously in the Middle East. The isolation of mycoplasmas is considered as one of the more difficult tasks for the diagnostic laboratories due to the inability of mycoplasmas to grow easily in laboratory media. The diagnosis of mycoplasma diseases is complex because clinical signs are not characteristic and antibody response can be transient or delayed in the early phase and non-existent in the final stages. Mycoplasma infections are notoriously difficult to prevent and control, rarely responding well to chemotherapy. Many countries control mycoplasma infections by slaughtering the diseased animals, but this way of control is impractical in the Middle East and especially in Jordan where there are limited funds for compensation. Vaccines may be a more efficient way of preventing the spread of disease but are not widely available and those inactivated mycoplasma vaccines that are available may provide only transient protection.

LESIONS ASSOCIATED WITH A NOVEL MYCOPLASMA SP. IN CALIFORNIA SEA LIONS (ZALOPHUS CALIFORNIANUS) UNDERGOING REHABILITATION

From July 1999 to November 2001, Mycoplasma sp. was cultured from lesions in 16 California sea lions (Zalophus californianus) undergoing rehabilitation. The Mycoplasma sp. was the likely cause of death of four animals in which it was associated with either pneumonia or polyarthritis. The most common lesion associated with this bacterium was subdermal abscessation, found in 12 animals. Other lesions included intramuscular abscesses, septic arthritis, and lymphadenopathy. Infection was associated with a leukocytosis and left shift in 12 animals. Animals with abscesses improved clinically after surgical lancing, irrigation, and systemic antibiotic therapy. The mycoplasma isolates had a consistent 16S rRNA sequence dissimilar from other Mycoplasma spp. and represent a novel species, Mycoplasma zalophi proposed sp. nov.

Pulsed-field gel electrophoresis patterns of Mycoplasma isolates from various body sites in dairy cattle with Mycoplasma mastitis

To determine whether Mycoplasma strains typically associated with mastitis in dairy cattle can be isolated from body sites other than the mammary gland. Prospective clinical trial. 7 Holstein cows in various stages of lactation with intramammary Mycoplasma infection. Milk samples, antemortem swab specimens from various body sites, and postmortem swab and tissue specimens were submitted for Mycoplasma culture. Pulsed-field gel electrophoresis (PFGE) was performed on chromosomal digests of all Mycoplasma isolates. Isolates with the same number and size of chromosomal digest bands were considered to be of the same type. For each cow, all isolates obtained from milk, mammary gland parenchyma, and supramammary lymph nodes had the same PFGE pattern. All cows had at least 1 isolate from nonmammary system tissues that had the same PFGE pattern as isolates from the mammary system. Overall, 44 of the 70 (63%) Mycoplasma isolates obtained from body sites other than mammary system sites had the same PFGE pattern as did mammary system isolates. Results confirmed our hypothesis that Mycoplasma strains isolated from the milk of dairy cattle with Mycoplasma mastitis frequently have PFGE patterns identical to those for strains isolated from other body sites, suggesting that there is at least a potential for internal transmission of Mycoplasma organisms.