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Sample records for mycoplasma mycoplasma gallisepticum

  1. Mycoplasma gallisepticum invades chicken erythrocytes during infection.

    PubMed

    Vogl, Gunther; Plaickner, Astrid; Szathmary, Susan; Stipkovits, László; Rosengarten, Renate; Szostak, Michael P

    2008-01-01

    Recently, it was demonstrated using in vitro assays that the avian pathogen Mycoplasma gallisepticum is able to invade nonphagocytic cells. It was also shown that this mycoplasma can survive and multiply intracellularly for at least 48 h and that this cell invasion capacity contributes to the systemic spread of M. gallisepticum from the respiratory tract to the inner organs. Using the gentamicin invasion assay and a differential immunofluorescence technique combined with confocal laser scanning microscopy, we were able to demonstrate in in vitro experiments that M. gallisepticum is also capable of invading sheep and chicken erythrocytes. The frequencies of invasion of three well-defined M. gallisepticum strains were examined over a period of 24 h, and a significant increase in invasiveness occurred after 8 h of infection. In addition, blood samples derived from chickens experimentally infected via the aerosol route with the virulent strain M. gallisepticum R(low) were analyzed. Surprisingly, M. gallisepticum R(low) was detected in the bloodstream of infected chickens by nested PCR, as well as by differential immunofluorescence and interference contrast microscopy that showed that mycoplasmas were not only on the surface but also inside chicken erythrocytes. This finding provides novel insight into the pathomechanism of M. gallisepticum and may have implications for the development of preventive strategies.

  2. Mycoplasma gallisepticum: Control by live attenuated vaccines

    USDA-ARS?s Scientific Manuscript database

    Commercially available attenuated strains of Mycoplasma gallisepticum (MG) are commonly used within the layer industry to control MG-induced mycoplasmosis. Among these are two live MG vaccines derived from the moderately pathogenic MG “chick F” strain. In the present study, the commercially availa...

  3. An epornitic of Mycoplasma gallisepticum in turkeys.

    PubMed

    Mason, S J; Maiers, J D

    1984-01-01

    A major epornitic of Mycoplasma gallisepticum occurred in the Monroe, North Carolina, area between January and June of 1983. The outbreak involved 304,000 turkeys of various ages, which were slaughtered in the eradication program at a cost of more than $550,000 to growers and poultry companies. An infected peafowl was the likely source of infection on the first farm. Traffic between farms by growers and company personnel was theorized to be the means of further spread.

  4. Use of polymerase chain reactions to detect Mycoplasma gallisepticum, Mycoplasma imitans, Mycoplasma iowae, Mycoplasma meleagridis and Mycoplasma synoviae in birds of prey.

    PubMed

    Lierz, M; Hagen, N; Lueschow, D; Hafez, H M

    2008-10-01

    Certain Mycoplasma spp. are pathogens of poultry, but little is known of the role of mycoplasmas in disease of birds of prey. Species-specific polymerase chain reactions (PCRs) for the detection of the poultry pathogens Mycoplasma gallisepticum, Mycoplasma imitans, Mycoplasma iowae, Mycoplasma meleagridis and Mycoplasma synoviae were therefore evaluated for use in birds of prey. The specificities of the PCR methods were established using avian and other mycoplasmas and also selected walled bacteria. The sensitivities of the different PCR assays varied between 100 fg and 10 pg DNA. Fifty-three tracheal swabs from healthy captive and free-ranging birds of prey were then investigated using these PCRs, and in no case was an amplicon obtained for M. gallisepticum/M. imitans, M. iowae or M. synoviae. Species-specific primers for M. meleagridis amplified a product from eight birds of prey but restriction enzyme analysis as well as sequencing of PCR products demonstrated these results to be false positives. Alignment studies of the sequenced products with the 16S rRNA gene sequence of various Mycoplasma species in GenBank demonstrated an identity of 91% to M. meleagridis but of 98% to Mycoplasma buteonis or Mycoplasma gallopavonis. Isolation and attempted identification of these mycoplasmas suggested it may be a previously unrecognized species.

  5. Attachment of killed Mycoplasma gallisepticum cells and membranes to erythrocytes

    SciTech Connect

    Banai, M.; Kahane, I.; Feldner, J.; Razin, S.

    1981-11-01

    To correlate viability with attachment capacity, Mycoplasma gallisepticum cells harvested at different growth phases and treated by various agents were tested for their capacity to attach to human erythrocytes. The results show that viability per se is not essential for M. gallisepticum attachment to erythrocytes, as cells killed by ultraviolet irradiation and membranes isolated by lysing M. gallisepticum cells by various means retained attachment capacity. However, treatment of the mycoplasmas by protein-denaturing agents, such as heart, glutaraldehyde, or prolonged exposure to low pH, drastically affected or even abolished attachment, supporting the protein nature of the mycoplasma membrane components responsible for specific binding to the sialoglycoprotein receptors on the erythrocytes.

  6. COLONIAL GROWTH OF MYCOPLASMA GALLISEPTICUM OBSERVED WITH THE ELECTRON MICROSCOPE

    PubMed Central

    Shifrine, Moshe; Pangborn, Jack; Adler, Henry E.

    1962-01-01

    Shifrine, Moshe (University of California, Davis), Jack Pangborn, and Henry E. Adler. Colonial growth of Mycoplasma gallisepticum observed with the electron microscope. J. Bacteriol. 83:187–192. 1962.—Mycoplasma gallisepticum strain S6 was grown on collodion film on solid medium. Samples were removed every few hours, fixed, washed, shadowed, and observed with the electron microscope. Three distinct forms of growth were observed: elementary cells (hexagonally shaped), platycytes, and exoblasts. A tentative mode of growth was postulated. The significance of the angular morphology to the relation between mycoplasmas and L-forms of bacteria is discussed. Images PMID:13911868

  7. The polymerase chain reaction for Mycoplasma gallisepticum detection.

    PubMed

    Kempf, I; Blanchard, A; Gesbert, F; Guittet, M; Bennejean, G

    1993-12-01

    On the basis of the aligned 16S rRNA sequences of Mollicutes, a pair of primers was chosen for the detection of Mycoplasma gallisepticum. When used in the polymerase chain reaction (PCR), the primers detected a specific amplification of all Mg strains tested, yielding an expected 330 bp product. Amplification was not detected when other Mollicutes or E. coli were used as PCR templates. SPF chickens were experimentally inoculated with two strains of M. gallisepticum or Mycoplasma iowae. Tracheal swabs were collected 8, 15, 20 and 28 days after inoculation, and cultured for mycoplasma or tested by PCR. PCR products were detected by hybridization with a digoxigenin-labeled probe and by chemiluminescence. The results showed that culture was positive for 49/73 swabs while PCR detected 70/72 positive samples. Thus, PCR can provide the basis of a sensitive, specific, rapid and non-radio-active method for detecting M. gallisepticum.

  8. Mechanisms of volume regulation in Mycoplasma gallisepticum

    SciTech Connect

    Linker, C.S.

    1987-01-01

    Mycoplasma gallisepticum, a cell wall-less bacterium, must confront the problem of colloid osmotic swelling. Cell volume was determined by optical density and intracellular water measurements. Transmembrane pH and electrical gradients were determined by the distribution of the weak acid benzoate and lipophilic cation tetraphenylphosphonium respectively. Cells incubated in sodium chloride without glucose exhibited a progressive fall in ATP over several hours. When ATP fell below 40 uM the cells swelled, leaked protein and became permeable to inulin. Subsequent addition of glucose induced shrinkage and restored the original permeability properties. Energized cells exhibited an electrochemical gradient of protons of up to 130 mV, inside negative and alkaline. The proton-ATPase inhibitor dicyclohexylcarbodiimide (DCCD), which collapsed the chemical and electrical components of the proton gradient, induced rapid swelling despite high ATP levels thus implicating the proton gradient in volume regulation. Either the pH gradient or the membrane potential could maintain volume. Energy-dependent sodium efflux in exchange for protons was demonstrated in sodium-loaded cells using radioactive sodium and 9-aminoacridine fluorescence to follow sodium and proton translocation respectively.

  9. Stability of rehydrated Mycoplasma gallisepticum vaccine homogeneity over time

    USDA-ARS?s Scientific Manuscript database

    Proper vaccine application is required to maximize the results of the vaccination, with maintenance of a homogenous solution is critical to obtain uniform results. This study was designed to analyze the need for continued mixing of a Mycoplasma gallisepticum vaccine solution in order to maintain a ...

  10. Proteomic analysis of Mycoplasma gallisepticum vaccine strain F

    USDA-ARS?s Scientific Manuscript database

    The persistence and displacement abilities of the Mycoplasma gallisepticum vaccine strain F (F-strain) are well documented. Understanding the mechanism(s) of colonization and persistence of F-strain will aid in the current intervention strategies to diagnose and control MG infections in poultry. In ...

  11. First identification of proteins involved in motility of Mycoplasma gallisepticum.

    PubMed

    Indikova, Ivana; Vronka, Martin; Szostak, Michael P

    2014-10-17

    Mycoplasma gallisepticum, the most pathogenic mycoplasma in poultry, is able to glide over solid surfaces. Although this gliding motility was first observed in 1968, no specific protein has yet been shown to be involved in gliding. We examined M. gallisepticum strains and clonal variants for motility and found that the cytadherence proteins GapA and CrmA were required for gliding. Loss of GapA or CrmA resulted in the loss of motility and hemadsorption and led to drastic changes in the characteristic flask-shape of the cells. To identify further genes involved in motility, a transposon mutant library of M. gallisepticum was generated and screened for motility-deficient mutants, using a screening assay based on colony morphology. Motility-deficient mutants had transposon insertions in gapA and the neighbouring downstream gene crmA. In addition, insertions were seen in gene mgc2, immediately upstream of gapA, in two motility-deficient mutants. In contrast to the GapA/CrmA mutants, the mgc2 motility mutants still possessed the ability to hemadsorb. Complementation of these mutants with a mgc2-hexahistidine fusion gene restored the motile phenotype. This is the first report assigning specific M. gallisepticum proteins to involvement in gliding motility.

  12. The development and application of a Mycoplasma gallisepticum sequence database.

    PubMed

    Armour, Natalie K; Laibinis, Victoria A; Collett, Stephen R; Ferguson-Noel, Naola

    2013-01-01

    Molecular analysis was conducted on 36 Mycoplasma gallisepticum DNA extracts from tracheal swab samples of commercial poultry in seven South African provinces between 2009 and 2012. Twelve unique M. gallisepticum genotypes were identified by polymerase chain reaction and sequence analysis of the 16S-23S rRNA intergenic spacer region (IGSR), M. gallisepticum cytadhesin 2 (mgc2), MGA_0319 and gapA genetic regions. The DNA sequences of these genotypes were distinct from those of M. gallisepticum isolates in a database composed of sequences from other countries, vaccine and reference strains. The most prevalent genotype (SA-WT#7) was detected in samples from commercial broilers, broiler breeders and layers in five provinces. South African M. gallisepticum sequences were more similar to those of the live vaccines commercially available in South Africa, but were distinct from that of F strain vaccine, which is not registered for use in South Africa. The IGSR, mgc2 or MGA_0319 sequences of three South African genotypes were identical to those of the ts-11 vaccine strain, necessitating a combination of mgc2 and IGSR targeted sequencing to differentiate South African wild-type genotypes from ts-11 vaccine. To identify and differentiate all 12 wild-types, mgc2, IGSR and MGA_0319 sequencing was required. Sequencing of gapA was least effective at strain differentiation. This research serves as a model for the development of an M. gallisepticum sequence database, and illustrates its application to characterize M. gallisepticum genotypes, select diagnostic tests and better understand the epidemiology of M. gallisepticum.

  13. Biological characterization of Russian Mycoplasma gallisepticum field isolates.

    PubMed

    Sprygin, Alexander V; Elatkin, Nikolay P; Kolotilov, Alexander N; Volkov, Mikhail S; Sorokina, Marina I; Borisova, Asya V; Andreychuk, Dmitriy B; Mudrak, Nataliya S; Irza, Victor N; Borisov, Alexander V; Drygin, Vladimir V

    2011-04-01

    An earlier study on commercial chickens and turkeys with a history of respiratory disease established Mycoplasma gallisepticum infection rates on 164 poultry farms of the Russian Federation. Forty-seven (29%) of these poultry farms were M. gallisepticum-positive by polymerase chain reaction but isolation of the mycoplasma was successful only on 10 farms. Five field isolates from different farms were selected for pathogenicity studies in specific pathogen-free chicks. Clinical signs, seroconversion, culture rates, air sac and tracheal lesions and mean tracheal mucosal thickness were all assessed in comparison with the reference strain, S6. Of the five isolates, MG140905 and MG070607 appeared to be slightly more pathogenic than the other three, as indicated by clinical signs, culture-positive rates and lesions, but only isolate MG140905 differed statistically (P < 0.05) from them, thus proving to be the most pathogenic. However, none of the Russian field isolates was as pathogenic as the S6 strain by the parameters measured. Stress or other factors such as concurrent bacterial or viral infections may have served as exacerbating factors for the disease seen in the naturally affected flocks. Sequence analysis of the gapA and mgc2 genes showed that MG140905 clustered with M. gallisepticum R(low) and was more distant from the majority of the Russian isolates.

  14. Mycoplasma gallisepticum infection in chukar partridges, pheasants, and peafowl.

    PubMed

    Cookson, K C; Shivaprasad, H L

    1994-01-01

    Mycoplasma gallisepticum infection was diagnosed in a group of chukar partridges, pheasants, and peafowl based on serology and isolation techniques. The farm also had quail, chickens, and ducks. Clinical signs in growing birds consisted of foamy eyes, swollen infraorbital sinuses, respiratory distress, and death. Breeding birds experienced a severe drop in egg production. Histologically, the growing birds exhibited lymphoplasmacytic inflammation of the conjunctiva, sinus, and trachea. The most likely source of infection was either chickens, which had been introduced before the onset of clinical signs, or the chukar partridge breeders, which had been obtained at various hunting field trials.

  15. Spray application of live attenuated F Strain-derived Mycoplasma gallisepticum vaccines

    USDA-ARS?s Scientific Manuscript database

    Live attenuated vaccines (LAVs) are commonly utilized to protect commercial table egg producers from economic losses associated with challenges by the respiratory pathogen Mycoplasma gallisepticum (MG). Currently there are four MG LAVs commercially available within the United States. Consistent am...

  16. Immunogenicity of a temperature sensitive mutant Mycoplasma gallisepticum vaccine.

    PubMed

    Whithear, K G; Soeripto; Harrigan, K E; Ghiocas, E

    1990-05-01

    The immunogenicity of the ts-11 vaccine strain of Mycoplasma gallisepticum was assessed following eye drop or coarse aerosol administration in chickens of various ages. Protection was evalualted following intra-abdominal (IA) or fine droplet aerosol administration of virulent M. gallisepticum, usually the Ap3AS strain and was measured mainly by the scoring of gross air sac lesions or by egg production. Vaccination of chickens with ts-11 did not elicit a substantial serum antibody response as measured by rapid serum agglutination test, or ELISA. Protection was never demonstrated when no M. gallisepticum serum antibody response was detected in a vaccinated group of chickens. Failure to protect occurred usually, although not invariably, following aerosol administration of the vaccine. Vaccination by eye drop usually, although not invariably provided protection against challenge. In one experiment, chickens vaccinated by eye drop at 8-weeks were as susceptible as non vaccinated controls when challenged by IA inoculation at 13-weeks-of-age. Yet other birds from the same vaccinated group were resistant when challenged in an identical way at 23-weeks. No measurable increase in M. gallisepticum specific serum antibody concentrations occurred in the intervening period. Equally surprising was the response of another group of birds in the same experiment that had been vaccinated with a higher dose of ts-11. An antibody response was detected in this group, but they were susceptible to challenge at 23-weeks. Interestingly, a drop in egg production commenced 4 weeks after challenge, 2 weeks later than that observed in a non vaccinated group challenged at the same time.(ABSTRACT TRUNCATED AT 250 WORDS)

  17. The PK/PD Interactions of Doxycycline against Mycoplasma gallisepticum

    PubMed Central

    Zhang, Nan; Gu, Xiaoyan; Ye, Xiaomei; Wu, Xun; Zhang, Bingxu; Zhang, Longfei; Shen, Xiangguang; Jiang, Hongxia; Ding, Huanzhong

    2016-01-01

    Mycoplasma gallisepticum is one of the most important pathogens that cause chronic respiratory disease in chicken. This study investigated the antibacterial activity of doxycycline against M. gallisepticum strain S6. In static time–killing studies with constant antibiotic concentrations [0–64 minimum inhibitory concentration (MIC)], M. gallisepticum colonies were quantified and kill rates were calculated to estimate the drug effect. The half-life of doxycycline in chicken was 6.51 ± 0.63 h. An in vitro dynamic model (the drug concentrations are fluctuant) was also established and two half-lives of 6.51 and 12 h were simulated. The samples were collected for drug concentration determination and viable counting of M. gallisepticum. In static time–killing studies, doxycycline produced a maximum antimycoplasmal effect of 5.62log10 (CFU/mL) reduction and the maximum kill rate was 0.11 h−1. In the in vitro dynamic model, doxycycline had a mycoplasmacidal activity in the two regimens, and the maximum antimycoplasmal effects were 4.1 and 4.75log10 (CFU/mL) reduction, respectively. Furthermore, the cumulative percentage of time over a 48-h period that the drug concentration exceeds the MIC (%T > MIC) was the pharmacokinetic–pharmacodynamic index that best correlated with antimicrobial efficacy (R2 = 0.986, compared with 0.897 for the peak level divided by the MIC and 0.953 for the area under the concentration–time curve over 48 h divided by the MIC). The estimated %T > MIC values for 0log10 (CFU/mL) reduction, 2log10 (CFU/mL) reduction and 3log10 (CFU/mL) reduction were 32.48, 45.68, and 54.36%, respectively, during 48 h treatment period of doxycycline. In conclusion, doxycycline shows excellent effectiveness and time-dependent characteristics against M. gallisepticum strain S6 in vitro. Additionally, these results will guide optimal dosing strategies of doxycycline in M. gallisepticum infection. PMID:27199972

  18. Mycoplasma gallisepticum: Influence of cell invasiveness on the outcome of experimental infection in chickens.

    PubMed

    Much, Peter; Winner, Florian; Stipkovits, László; Rosengarten, Renate; Citti, Christine

    2002-11-15

    Recently we have shown that a low (R(low)) and a high laboratory passage (R(high)) of the poultry pathogen Mycoplasma gallisepticum prototype strain R differ markedly in their capability to invade non-phagocytic eukaryotic cells. In the present study the infection traits of these two mycoplasma passages were compared in an in vivo setting. After aerosol inoculation of chickens, M. gallisepticum was re-isolated from the inner organs of birds infected with R(low), whereas no mycoplasma was recovered from the inner organs of birds infected with R(high). These results indicate that the two mycoplasma populations derived from strain R differ in their capacity to cross the mucosal barrier and suggest that cell invasion may play a major role in the observed systemic spreading of M. gallisepticum in its chicken host.

  19. In vitro susceptibilities to fluoroquinolones in current and archived Mycoplasma gallisepticum and Mycoplasma synoviae isolates from meat-type turkeys.

    PubMed

    Gerchman, Irina; Lysnyansky, Inna; Perk, Shimon; Levisohn, Sharon

    2008-10-15

    Monitoring of susceptibility to antibiotics in field isolates of pathogenic avian mycoplasmas is important for appropriate choice of treatment. Our study compared in vitro susceptibility to enrofloxacin and difloxacin in recent (2005-2006) isolates of Mycoplasma gallisepticum and Mycoplasma synoviae from meat-type turkey flocks with archived (1997-2003) isolates and reference strains. Comparison of minimal inhibitory concentration (MIC) values determined by microtest, agar dilution and commercial Etest showed good agreement, but underscored the need for standardized methods for testing. Notably, while the commercial Etest was convenient and accurate for determining MICs for enrofloxacin in the range 0.002-0.094microg/ml, the endpoint of inhibition for M. gallisepticum and M. synoviae strains with MIC values > or =1.0microg/ml could not be determined. A decrease in susceptibility to both fluoroquinolones was detected in archived strains but to a greater degree in recent isolates, most of which had MICs above the NCCLS susceptibility breakpoint for these antibiotics (< or =0.5microg/ml). In contrast, except for one flock, M. synoviae isolates were susceptible, although intrinsically less susceptible than M. gallisepticum. Overall for the 88 strains tested (45 M. gallisepticum, 43 M. synoviae), the MIC50 for both enrofloxacin and difloxacin was 0.5microg/ml. The isolation of fluoroquinolone-resistant M. gallisepticum isolates from breeder and broiler flocks as well as from meat-type turkeys suggests that these strains have become established in Israel, necessitating a reevaluation of antibiotic therapy. Periodic survey of MICs in field isolates of avian mycoplasmas to monitor for the possible appearance of resistant strains is recommended.

  20. Effects of prelay ts11-strain Mycoplasma gallisepticum inoculation and time specific F-strain Mycoplasma gallisepticum inoculation overlays on internal egg and eggshell characteristics of commercial laying hens

    USDA-ARS?s Scientific Manuscript database

    Mycoplasma infections are pandemic in multiage layer chicken flocks with M. gallisepticum being the species of greatest concern to commercial egg producers. Live M. gallisepticum vaccines are presently being used to help control M. gallisepticum outbreaks. However, vaccination of layers with F-str...

  1. Change in antimicrobial susceptibility of Mycoplasma gallisepticum field isolates.

    PubMed

    Gharaibeh, Saad; Al-Rashdan, Mohammad

    2011-06-02

    This study compares the antimicrobial susceptibility over time between two groups of Mycoplasma gallisepticum (MG) isolates from the same geographical area. Minimum inhibitory concentration of 13 antimicrobials was determined against two groups of MG isolates from chickens. Group 1 strains (n=22) were isolated in 2004-2005 while group 2 strains (n=7) were isolated in 2007-2008. Minimum inhibitory concentration 50 for group 1 versus group 2 was 4/4, 0.5/0.5, ≤ 0.031/≥ 64, ≤ 0.031/2, ≤ 0.031/0.125, 1/0.5, 1/1, ≤ 0.031/≤ 0.031, ≤ 0.031/2, ≤ 0.031/2, 1/4, ≤ 0.031/0.062, and 0.062/2 μg/ml against gentamicin, spectinomycin, erythromycin, tilmicosin, tylosin, florfenicol, thiamphenicol, tiamulin, ciprofloxacin, enrofloxacin, chlortetracycline, doxycycline, and oxytetracycline, respectively. There was a statistically significant increase in resistance of group 2 to erythromycin, tilmicosin, tylosin, ciprofloxacin, enrofloxacin, chlortetracycline, doxycycline, and oxytetracycline. This dramatic increase in resistance against 8 antimicrobials belonging to three different families of antimicrobials in a relatively short period of time appears to be rare and of concern. The cause of this increased resistance observed in group 2 of MG isolates was not determined and should be further investigated. Monitoring of MG field strain susceptibility is highly recommended to implement successful treatment and prophylaxis programs in endemic areas. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. Experimental Mycoplasma gallisepticum infections in captive-reared wild turkeys

    USGS Publications Warehouse

    Rocke, Tonie E.; Yuill, Thomas M.; Amundson, Terry E.

    1988-01-01

    The effects of Mycoplasma gallisepticum (MG) infections on egg production, fertility, and hatchability were studied in captive-reared wild turkeys (Meleagris gallopavo). Three groups of adult birds, each consisting of four hens and two toms, were exposed to MG by the respiratory route at the beginning of their breeding season. Fourteen control birds received sterile growth medium. Although no mortality of infected or control birds occurred, egg production during the first breeding season after infection was reduced. The mean number of eggs/hen/day produced by infected groups the first breeding season postexposure (PE) was significantly lower than the control value. The mean number of eggs produced daily by the same hens 1 yr later was unaffected by MG infection. The pecentage of fertile eggs produced by infected groups was slightly reduced in both the first and second breeding seasons PE. Hatchability of fertile eggs from infected hens was significantly lower than eggs from control hens. Productivity may be impaired if MG infections occur in free-ranging wild turkey populations.

  3. Effect of infection route and concurrent infectious bronchitis virus vaccination on Mycoplasma gallisepticum disease pathology in an experimental model

    USDA-ARS?s Scientific Manuscript database

    The study of Mycoplasma gallisepticum infections is needed, not only to understand the disease process, but also to understand the mechanisms by which M. gallisepticum vaccines protect the host. Many model systems have been used to study the M. gallisepticum disease process. This work compared two...

  4. Pharmacokinetic/Pharmacodynamic Profiles of Tiamulin in an Experimental Intratracheal Infection Model of Mycoplasma gallisepticum

    PubMed Central

    Xiao, Xia; Sun, Jian; Yang, Tao; Fang, Xi; Cheng, Jie; Xiong, Yan Q.; Liu, Ya-Hong

    2016-01-01

    Mycoplasma gallisepticum is the most important pathogen in poultry among four pathogenic Mycoplasma species. Tiamulin is a pleuromutilin antibiotic that shows a great activity against M. gallisepticum and has been approved for use in veterinary medicine particularly for poultry. However, the pharmacokinetic/pharmacodynamics (PK/PD) profiles of tiamulin against M. gallisepticum are not well understood. Therefore, in the current studies, we investigated the in vivo PK/PD profiles of tiamulin using a well-established experimental intratracheal infection model of M. gallisepticum. The efficacy of tiamulin against M. gallisepticum was studied in 8-day-old chickens after intramuscular (i.m.) administration at 10 doses between 0–80 mg/kg. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to evaluate the PK parameters of tiamulin following i.m. administration at doses of 5, 40, and 80 mg/kg in Mycoplasma gallisepticum-infected neutropenic chickens. Real-time PCR (RT-PCR) was used for quantitative detection of M. gallisepticum. The MIC of tiamulin against M. gallisepticum strain S6 was 0.03 μg/mL. The PK/PD index, AUC24h/MIC, correlated well with the in vivo antibacterial efficacy. The in vivo data suggest that animal dosage regimens should supply AUC24h/MIC of tiamulin of 382.68 h for 2 log10 ccu equivalents M. gallisepticum reduction. To attain that goal, the administered dose is expected to be 45 mg/kg b.w. for treatment of M. gallisepticum infection with an MIC90 of 0.03 μg/mL. PMID:27656647

  5. Effects of different vaccine combinations against Mycoplasma gallisepticum on blood characteristics in commercial layer chickens

    USDA-ARS?s Scientific Manuscript database

    Mycoplasma gallisepticum (MG) is a major and economically significant pathogen of avian species. When administered before lay, F-strain MG (FMG) can reduce egg production during lay, but the ts-11 strain of MG (ts11MG) does not exert this effect. Two trials were conducted to determine the effects ...

  6. A comparative study of live attenuated F strain-derived Mycoplasma gallisepticum vaccines

    USDA-ARS?s Scientific Manuscript database

    Commercially available attenuated strains of Mycoplasma gallisepticum (MG) are commonly used within the layer industry to control MG-induced mycoplasmosis. Among these are two live MG vaccines derived from the moderately pathogenic MG “chick F” strain. In the present study, the commercially availa...

  7. Transformation of Mycoplasma gallisepticum with Tn916, Tn4001, and integrative plasmid vectors.

    PubMed Central

    Cao, J.; Kapke, P. A.; Minion, F. C.

    1994-01-01

    Mycoplasma gallisepticum causes respiratory disease in avian species, but little is known about its mechanism(s) of pathogenesis. These studies were undertaken in order to develop genetic systems for analysis of potential virulence factors. M. gallisepticum was transformed with plasmids containing one of the gram-positive transposons Tn916 or Tn4001, which inserted randomly into the mycoplasmal chromosome. Plasmids containing cloned chromosomal DNA were also constructed and tested for integration into regions of DNA homology derived either from chromosomal fragments or from the gentamicin resistance marker from Tn4001. These studies demonstrate that M. gallisepticum is amenable to transformation with both transposons and integrative vectors. Images PMID:8021232

  8. A surface epitope undergoing high-frequency phase variation is shared by Mycoplasma gallisepticum and Mycoplasma bovis.

    PubMed Central

    Yogev, D; Menaker, D; Strutzberg, K; Levisohn, S; Kirchhoff, H; Hinz, K H; Rosengarten, R

    1994-01-01

    We have recently reported that three distinct size- and phase-variable surface lipoproteins (Vsps) of the bovine pathogen Mycoplasma bovis possess a common epitope recognized by monoclonal antibody 1E5. In the present study, we show that this epitope is also present on a size-variant protein (PvpA) of the avian pathogen Mycoplasma gallisepticum. Application of monoclonal antibody 1E5 in Western immunoblot analysis of Triton X-114 phase-fractionated proteins and in colony immunoblots, as well as in trypsin and carboxypeptidase digestion experiments, has demonstrated that (i) PvpA is an integral membrane protein with a free C terminus, (ii) the shared epitope is surface exposed, and (iii) PvpA is subjected to high-frequency phase variation in expression. By using serum antibodies from M. gallisepticum-infected chickens, we were able to demonstrate the immunogenic nature of PvpA and identify three additional highly immunogenic Triton X-114 phase proteins (p67, p72, and p75) also undergoing high-frequency phase variation spontaneously and independently. Metabolic labeling experiments with [14C]palmitate and [14C]oleate revealed that PvpA, in contrast to p67, p72, and p75, is not lipid modified. Southern blot hybridization with restriction fragments carrying the pvpA gene of M. gallisepticum or the vspA gene of M. bovis against digested genomic DNA of the two Mycoplasma species indicated the absence of genetic relatedness between the pvpA and vspA genes. The apparent complexity of the antigenic variation phenomenon in M. gallisepticum is discussed. Images PMID:7523302

  9. A chronicle of serologic response in commercial layer chickens to vaccination with commercial F strain Mycoplasma gallisepticum vaccine

    USDA-ARS?s Scientific Manuscript database

    Initial efforts by the poultry industry at controlling and containing Mycoplasma gallisepticum (MG) included testing and slaughter of reactor flocks. Ultimately, using the aforementioned measures coupled with heat treatment of hatching eggs together with biosecurity and biosurveillance procedures, ...

  10. Effect of selected water temperatures used in Mycoplasma gallisepticum vaccine reconstitution on titer at selected time intervals

    USDA-ARS?s Scientific Manuscript database

    Numerous methods are currently used throughout the poultry industry for the administration of vaccines. Each utilizes water for vaccine reconstitution and/or administration, including two of the three commercially available live Mycoplasma gallisepticum (MG) vaccines. Selected water temperatures w...

  11. Serological Screening Suggests Extensive Presence of Mycoplasma gallisepticum and Mycoplasma synoviae in Backyard Chickens in Southern Mozambique

    PubMed Central

    Taunde, Paula; Zandamela, Ana Felicidade; Junior, Alberto Pondja; Chilundo, Abel; Costa, Rosa

    2017-01-01

    A total of 459 serum samples from unvaccinated backyard chickens originating from 4 villages in Mandlakazi district, Southern Mozambique, were tested for the presence of Mycoplasma gallisepticum and Mycoplasma synoviae antibodies through commercial enzyme-linked immunoabsorbent assay [ELISA] kits. Anti-MG and anti-MS antibodies were detected in all villages surveyed and the overall seroprevalence was 48.8% [95% CI 39.1–57.8] and 84.5% [95% CI 76.8–90.4], respectively. The risk of being seropositive for both diseases was higher [P < 0.05] in Chidenguele village than other villages. It is concluded that MG and MS serum antibodies are present in backyard chickens. PMID:28243629

  12. Detection of infectious bronchitis virus 793B, avian metapneumovirus, Mycoplasma gallisepticum and Mycoplasma synoviae in poultry in Ethiopia.

    PubMed

    Hutton, S; Bettridge, J; Christley, R; Habte, T; Ganapathy, K

    2017-02-01

    A survey was conducted into respiratory infectious diseases of poultry on a chicken breeder farm run by the Ethiopian Institute of Agricultural Research (EIAR), located in Debre Zeit, Ethiopia. Oropharyngeal swabs were collected from 117 randomly selected birds, and blood was taken from a subset of 73 of these birds. A combination of serological and molecular methods was used for detection of pathogens. For the first time in Ethiopia, we report the detection of variant infectious bronchitis virus (793B genotype), avian metapneumovirus subtype B and Mycoplasma synoviae in poultry. Mycoplasma gallisepticum was also found to be present; however, infectious laryngotracheitis virus was not detected by PCR. Newcastle disease virus (NDV) was not detected by PCR, but variable levels of anti-NDV HI antibody titres shows possible exposure to virulent strains or poor vaccine take, or both. For the burgeoning-intensive industry in Ethiopia, this study highlights several circulating infectious respiratory pathogens that can impact on poultry welfare and productivity.

  13. Effects of Time-Specific F-strain Mycoplasma gallisepticum Inoculation Overlays on Prelay ts-11-strain M. gallisepticum Vaccination on Blood Characteristics of Commercial Laying Hens

    USDA-ARS?s Scientific Manuscript database

    Two trials were conducted to determine the effects of a prelay ts-11-strain Mycoplasma gallisepticum (ts-11MG) vaccination alone or in combination with subsequent time specific F-strain M. gallisepticum (FMG) inoculations on the blood characteristics of commercial laying hens. The following 4 treat...

  14. Effects of Prelay 6/85-Strain Mycoplasma gallisepticum Inoculation Alone or in Conjunction with the Inoculation of F-Strain Mycoplasma gallisepticum During Lay on the Blood Characteristics of Commercial Egg-Laying Hens

    USDA-ARS?s Scientific Manuscript database

    The effects of 6/85 Mycoplasma gallisepticum (6/85MG) inoculation alone or in conjunction with F-strain M. Gallisepticum (FMG) overlays and their timing on the blood characteristics of commercial egg-laying hens were investigated. Control birds received sham inoculations at 10 wk of age. Birds in ...

  15. Experimental infection of chickens with an atypical Mycoplasma gallisepticum strain: comparison of diagnostic methods.

    PubMed

    Kempf, I; Gesbert, F; Guittet, M

    1997-01-01

    Fifteen chickens were inoculated with the atypical Mycoplasma gallisepticum (MG) K703 strain. On different dates post inoculation, tracheal swab samples were collected for mycoplasma culture and blood samples were analysed by slide agglutination test (SA) with commercial or homologous antigen and enzyme-linked immunosorbent assay (ELISA) with three different kits. Results showed that MG isolation rate was low on several sampling dates. The SA with commercial antigen did not yield positive results, although birds were positive when tested with homologous antigen. With commercial ELISA kits, the numbers of positive samples remained low. These results illustrate the difficulty of diagnosis of infections with such MG variant strains.

  16. Characterisation of Mycoplasma gallisepticum strains involved in respiratory disease in pheasants and peafowl.

    PubMed

    Bencina, D; Mrzel, I; RoJs, O Zorman; Bidovec, A; Dovc, A

    2003-02-22

    Two cases of Mycoplasma gallisepticum infection in different avian species in backyard gamebird operations in Slovenia were investigated. In the first case, M gallisepticum was associated with severe respiratory disease with almost 20 per cent mortality in pheasants, whereas the infection was less pathogenic for chickens and turkeys reared at the same site. The M gallisepticum isolates from pheasants had a unique pMGA gene sequence containing a repeat of 12 nucleotides, and they contained only small amounts of the cytadhesins MGC1 and MGC3 and no PvpA protein. However, they expressed some typical M gallisepticum proteins and several proteins which were immunogenic for pheasants, chickens and turkeys. A strain of M gallisepticum isolated from the sinus of a pheasant was highly pathogenic for chicken embryos. In the second case, the M gallisepticum strain that was associated with respiratory disease and mortality in peafowl also affected chickens. M gallisepticum strain ULB 992 was isolated from the infraorbital sinus of a dead peafowl. The ULB 992 strain synthesised a small amount of MGC3, a truncated form of MGC1 and lacked PvpA. However, it expressed several proteins which were immunogenic for the birds infected with M gallisepticum at both gamebird operations.

  17. Development of a Site-Directed Integration Plasmid for Heterologous Gene Expression in Mycoplasma gallisepticum

    PubMed Central

    Indikova, Ivana; Szostak, Michael P.

    2013-01-01

    Deciphering the molecular basis of the interactions between the parasite Mycoplasma gallisepticum and its avian hosts suffers from the lack of genetic tools available for the pathogen. In the absence of well established methods for targeted disruption of relevant M. gallisepticum genes, we started to develop suicide vectors and equipped them with a short fragment of M. gallisepticum origin or replication (oriCMG). We failed to create a disruption vector, although by adding a further short fragment of the M. gallisepticum tufB upstream region we created a “Trojan horse” plasmid. This is fully integrated into the genomic DNA of M. gallisepticum, always at the same site, oriCMG, and is able to carry and express any gene of interest in the genetic background of M. gallisepticum. Successful expression of a heterologous gene was shown with the lacZ gene of E. coli. When used for gene complementation or expression of hybrid genes in M. gallisepticum, a site-specific combined integration/expression vector constitutes an improvement on randomly integrating transposons, which might have unexpected effects on the expression of chromosomal genes. PMID:24278444

  18. Identification of Strain-Specific Sequences That Distinguish a Mycoplasma gallisepticum Vaccine Strain from Field Isolates

    PubMed Central

    Ricketts, Camir; Pickler, Larissa; Maurer, John; Ayyampalayam, Saravanaraj; García, Maricarmen

    2016-01-01

    ABSTRACT Despite attempts to control avian mycoplasmosis through management, vaccination, and surveillance, Mycoplasma gallisepticum continues to cause significant morbidity, mortality, and economic losses in poultry production. Live attenuated vaccines are commonly used in the poultry industry to control avian mycoplasmosis; unfortunately, some vaccines may revert to virulence and vaccine strains are generally difficult to distinguish from natural field isolates. In order to identify genome differences among vaccine revertants, vaccine strains, and field isolates, whole-genome sequencing of the M. gallisepticum vaccine strain ts-11 and several “ts-11-like” strains isolated from commercial flocks was performed using Illumina and 454 pyrosequencing and the sequenced genomes compared to the M. gallisepticum Rlow reference genome. The collective contigs for each strain were annotated using the fully annotated Mycoplasma reference genome. The analysis revealed genetic differences among vlhA alleles, as well as among genes annotated as coding for a cell wall surface anchor protein (mg0377) and a hypothetical protein gene, mg0359, unique to M. gallisepticum ts-11 vaccine strain. PCR protocols were designed to target 5 sequences unique to the M. gallisepticum ts-11 strain: vlhA3.04a, vlhA3.04b, vlhA3.05, mg0377, and mg0359. All ts-11 isolates were positive for the five gene alleles tested by PCR; however, 5 to 36% of field isolates were also positive for at least one of the alleles tested. A combination of PCR tests for vlhA3.04a, vlhA3.05, and mg0359 was able to distinguish the M. gallisepticum ts-11 vaccine strain from field isolates. This method will further supplement current approaches to quickly distinguish M. gallisepticum vaccine strains from field isolates. PMID:27847370

  19. The Mycoplasma gallisepticum Virulence Factor Lipoprotein MslA Is a Novel Polynucleotide Binding Protein

    PubMed Central

    Masukagami, Yumiko; Tivendale, Kelly A.; Mardani, Karim; Ben-Barak, Idan; Markham, Philip F.

    2013-01-01

    Although lipoproteins of mycoplasmas are thought to play a crucial role in interactions with their hosts, very few have had their biochemical function defined. The gene encoding the lipoprotein MslA in Mycoplasma gallisepticum has recently been shown to be required for virulence, but the biochemical function of this gene is not known. Although this gene has no significant sequence similarity to any gene of known function, it is located within an operon in M. gallisepticum that contains a homolog of a gene previously shown to be a nonspecific exonuclease. We mutagenized both genes to facilitate expression in Escherichia coli and then examined the functions of the recombinant proteins. The capacity of MslA to bind polynucleotides was examined, and we found that the protein bound single- and double-stranded DNA, as well as single-stranded RNA, with a predicted binding site of greater than 1 nucleotide but less than or equal to 5 nucleotides in length. Recombinant MslA cleaved into two fragments in vitro, both of which were able to bind oligonucleotides. These findings suggest that the role of MslA may be to act in concert with the lipoprotein nuclease to generate nucleotides for transport into the mycoplasma cell, as the remaining genes in the operon are predicted to encode an ABC transporter. PMID:23798535

  20. Mycoplasma gallisepticum inactivated by targeting the hydrophobic domain of the membrane preserves surface lipoproteins and induces a strong immune response.

    PubMed

    Atalla, Hazem; Lysnyansky, Inna; Raviv, Yossef; Rottem, Shlomo

    2015-01-01

    An innovative approach for inactivation of Mycoplasma gallisepticum using the hydrophobic photoinduced alkylating probe 1, 5-iodonaphthylazide (INA) is described. Treatment of washed M. gallisepticum mid-exponential culture (0.2 mg cell protein /mL) with INA followed by irradiation with far-ultraviolet light (310-380 nm) completely abolished viability. Transmission electron microscopy showed that the majority of the inactivated M. gallisepticum were comparable in size to intact cells, but that part of the INA-treated M. gallisepticum preparation also contained low density cells and membrane vesicles. Confocal microscopy revealed that untreated M. gallisepticum cells were internalized by chicken red blood cells (c-RBCs), whereas the INA-inactivated cells remained attached to the outer surface of the c-RBCs. INA treatment of M. gallisepticum resulted in a complete inactivation of F0F1 -ATPase and of the L-arginine uptake system, but the cytoplasmatic soluble NADH2 dehydrogenase was only partially affected. Western blot analysis of the lipoprotein fraction showed that the INA-treated M. gallisepticum retained their lipoproteins. Following subcutaneous injection of M. gallisepticum INA-bacterin, 100% and 68.8% of chickens were positive by the rapid serum agglutination test and enzyme-linked immunosorbent assay respectively, 2 weeks post-injection. These data suggest that the photoinducible alkylating agent INA inactivates M. gallisepticum but preserves its surface lipoproteins and thus has the potential to be used as a general approach for the inactivation of mycoplasmas for vaccine development.

  1. GapA and CrmA Coexpression Is Essential for Mycoplasma gallisepticum Cytadherence and Virulence

    PubMed Central

    Papazisi, L.; Frasca Jr., S.; Gladd, M.; Liao, X.; Yogev, D.; Geary, S. J.

    2002-01-01

    It was previously demonstrated that avirulent Mycoplasma gallisepticum strain Rhigh (passage 164) is lacking three proteins that are expressed in its virulent progenitor, strain Rlow (passage 15). These proteins were identified as the cytadhesin molecule GapA, the putative cytadhesin-related molecule CrmA, and a component of a high-affinity transporter system, HatA. Complementation of Rhigh with wild-type gapA restored expression in the transformant (GT5) but did not restore the cytadherence phenotype and maintained avirulence in chickens. These results suggested that CrmA might play an essential role in the M. gallisepticum cytadherence process. CrmA is encoded by the second gene in the gapA operon and shares significant sequence homology to the ORF6 gene of Mycoplasma pneumoniae, which has been shown to play an accessory role in the cytadherence process. Complementation of Rhigh with wild-type crmA resulted in the transformant (SDCA) that lacked the cytadherence and virulence phenotype comparable to that found in Rhigh and GT5. In contrast, complementation of Rhigh with the entire wild-type gapA operon resulted in the transformant (GCA1) that restored cytadherence to the level found in wild-type Rlow. In vivo pathogenesis trials revealed that GCA1 had regained virulence, causing airsacculitis in chickens. These results demonstrate that both GapA and CrmA are required for M. gallisepticum cytadherence and pathogenesis. PMID:12438360

  2. Proteomic analysis of tylosin-resistant Mycoplasma gallisepticum reveals enzymatic activities associated with resistance.

    PubMed

    Xia, Xi; Wu, Congming; Cui, Yaowen; Kang, Mengjiao; Li, Xiaowei; Ding, Shuangyang; Shen, Jianzhong

    2015-11-20

    Mycoplasma gallisepticum is a significant pathogenic bacterium that infects poultry, causing chronic respiratory disease and sinusitis in chickens and turkeys, respectively. M. gallisepticum infection poses a substantial economic threat to the poultry industry, and this threat is made worse by the emergence of antibiotic-resistant strains. The mechanisms of resistance are often difficult to determine; for example, little is known about antibiotic resistance of M. gallisepticum at the proteome level. In this study, we performed comparative proteomic analyses of an antibiotic (tylosin)-resistant M. gallisepticum mutant and a susceptible parent strain using a combination of two-dimensional differential gel electrophoresis and nano-liquid chromatography-quadrupole-time of flight mass spectrometry. Thirteen proteins were identified as differentially expressed in the resistant strain compared to the susceptible strain. Most of these proteins were related to catalytic activity, including catalysis that promotes the formylation of initiator tRNA and energy production. Elongation factors Tu and G were over-expressed in the resistant strains, and this could promote the binding of tRNA to ribosomes and catalyze ribosomal translocation, the coordinated movement of tRNA, and conformational changes in the ribosome. Taken together, our results indicate that M. gallisepticum develops resistance to tylosin by regulating associated enzymatic activities.

  3. Proteomic analysis of tylosin-resistant Mycoplasma gallisepticum reveals enzymatic activities associated with resistance

    PubMed Central

    Xia, Xi; Wu, Congming; Cui, Yaowen; Kang, Mengjiao; Li, Xiaowei; Ding, Shuangyang; Shen, Jianzhong

    2015-01-01

    Mycoplasma gallisepticum is a significant pathogenic bacterium that infects poultry, causing chronic respiratory disease and sinusitis in chickens and turkeys, respectively. M. gallisepticum infection poses a substantial economic threat to the poultry industry, and this threat is made worse by the emergence of antibiotic-resistant strains. The mechanisms of resistance are often difficult to determine; for example, little is known about antibiotic resistance of M. gallisepticum at the proteome level. In this study, we performed comparative proteomic analyses of an antibiotic (tylosin)-resistant M. gallisepticum mutant and a susceptible parent strain using a combination of two-dimensional differential gel electrophoresis and nano-liquid chromatography-quadrupole-time of flight mass spectrometry. Thirteen proteins were identified as differentially expressed in the resistant strain compared to the susceptible strain. Most of these proteins were related to catalytic activity, including catalysis that promotes the formylation of initiator tRNA and energy production. Elongation factors Tu and G were over-expressed in the resistant strains, and this could promote the binding of tRNA to ribosomes and catalyze ribosomal translocation, the coordinated movement of tRNA, and conformational changes in the ribosome. Taken together, our results indicate that M. gallisepticum develops resistance to tylosin by regulating associated enzymatic activities. PMID:26584633

  4. Mycoplasma gallisepticum infection in drug-treated chickens: comparison of diagnosis methods including polymerase chain reaction.

    PubMed

    Kempf, I; Gesbert, F; Guittet, M; Bennejean, G

    1994-11-01

    Ten chickens were inoculated with Mycoplasma gallisepticum (MG) and treated with enrofloxacine. On eight different dates post-inoculation (PI), tracheal swab samples were collected for mycoplasma culture or detection by polymerase chain reaction (PCR), and blood samples were analysed by slide-agglutination test (SA) and enzyme-linked immunosorbent assay (ELISA). Results showed that culture and PCR detected MG from 14/80 or 20/80 samples, respectively. The last culture-positive sample was collected on day 26 PI, whereas PCR still gave positive results on day 54 PI. This difference may be attributed to the high sensitivity of PCR and to its ability to detect non-viable or non-culturable pathogens. Sera were SA positive as early as 5 days PI and some of them remained positive up to day 47 PI. ELISA detected 53 suspicious or positive sera.

  5. Testing the efficacy of fermented wheat germ extract against Mycoplasma gallisepticum infection of chickens.

    PubMed

    Stipkovits, L; Lapis, K; Hidvégi, M; Kósa, E; Glávits, R; Resetár, A

    2004-11-01

    The effect of fermented wheat germ extract (FWGE, Immunovet-HBM) was studied in chickens challenged with Mycoplasma gallisepticum. Ninety M. gallisepticum- and M. synoviae-free 3-wk-old chickens were exposed to aerosol infection of M. gallisepticum. One group (30 birds) was treated with FWGE, a second group with tiamulin, and a third group was untreated. The fourth group was exposed to PBS aerosol as a negative control. On d 9, all chickens were slaughtered and examined for the presence of gross and histological lesions, the presence of the challenge strain in the organs and specific antibodies in the serum. Body weight gains and feed conversion rates were recorded. In the groups treated with FWGE and with tiamulin, the chickens remained clinically healthy: their BW gains were 441.7 g and 446.8 g, respectively. Feed conversion ratios were 1.72 and 1.71 for FWGE- and tiamulin-treated birds, respectively. Control birds had BW gain of 480.8 g, and feed conversion ratio of 1.78. The numbers of birds with gross lesions (15 and 11, respectively) and lesion scores (25 and 25, respectively) of the FWGE- and tiamulin-treated groups were significantly lower than in the infected untreated group (25 birds, lesion score of 190). No mycoplasma was reisolated from brain, liver, spleen, heart, or kidneys of the FWGE-treated birds, and the number of mycoplasma isolations from the respiratory tract samples was less frequent (10) than from the infected untreated group (64). In addition, 35 samples from other internal organs were also positive. Twenty percent of the birds treated with FWGE showed serological response with a 5.0% reaction score, whereas in the infected untreated group, 83.3% of birds were reactors, with a 62.5% reaction score.

  6. Interaction of Mycoplasma gallisepticum with Chicken Tracheal Epithelial Cells Contributes to Macrophage Chemotaxis and Activation

    PubMed Central

    Majumder, Sanjukta

    2015-01-01

    Mycoplasma gallisepticum colonizes the chicken respiratory mucosa and mediates a severe inflammatory response hallmarked by subepithelial leukocyte infiltration. We recently reported that the interaction of M. gallisepticum with chicken tracheal epithelial cells (TECs) mediated the upregulation of chemokine and inflammatory cytokine genes in these cells (S. Majumder, F. Zappulla, and L. K. Silbart, PLoS One 9:e112796, http://dx.doi.org/10.1371/journal.pone.0112796). The current study extends these observations and sheds light on how this initial interaction may give rise to subsequent inflammatory events. Conditioned medium from TECs exposed to the virulent Rlow strain induced macrophage chemotaxis to a much higher degree than the nonvirulent Rhigh strain. Coculture of chicken macrophages (HD-11) with TECs exposed to live mycoplasma revealed the upregulation of several proinflammatory genes associated with macrophage activation, including interleukin-1β (IL-1β), IL-6, IL-8, CCL20, macrophage inflammatory protein 1β (MIP-1β), CXCL-13, and RANTES. The upregulation of these genes was similar to that observed upon direct contact of HD-11 cells with live M. gallisepticum. Coculture of macrophages with Rlow-exposed TECs also resulted in prolonged expression of chemokine genes, such as those encoding CXCL-13, MIP-1β, RANTES, and IL-8. Taken together, these studies support the notion that the initial interaction of M. gallisepticum with host respiratory epithelial cells contributes to macrophage chemotaxis and activation by virtue of robust upregulation of inflammatory cytokine and chemokine genes, thereby setting the stage for chronic tissue inflammation. PMID:26527215

  7. Effects of sialidase knockout and complementation on virulence of Mycoplasma gallisepticum.

    PubMed

    May, Meghan; Szczepanek, Steven M; Frasca, Salvatore; Gates, Amy E; Demcovitz, Dina L; Moneypenny, Craig G; Brown, Daniel R; Geary, Steven J

    2012-05-25

    Reannotation of the pathogenic Mycoplasma gallisepticum strain R(low) genome identified the hypothetical gene MGA_0329 as a homolog of the sialidase gene MS53_0199 of Mycoplasma synoviae strain MS53. Potent sialidase activity was subsequently quantitated in several M. gallisepticum strains. Because sialidase activity levels correlate significantly with differing M. synoviae strain virulence, we hypothesized this enzyme may also influence the virulence of M. gallisepticum. MGA_0329 was disrupted in strain R(low) to create mutants 6, 358 and P1C5, which resulted in the loss of sialidase activity in all three mutants. Chickens infected with the knockout mutants had significantly less severe (P<0.05) tracheal lesions and tracheal mucosal thickening than chickens infected with equal doses of strain R(low). Significantly fewer (P<0.05) CCU especially of strains 6 and P1C5 were recovered at necropsy. Mini-Tn4001tet plasmid pTF20 carrying a wild-type copy of MGA_0329 with its native promoter was used to complement the genetic lesion in strain P1C5. Three clones derived from P1C5, each having one copy of MGA_0329 stably transposed into a different site in its genome, expressed sialidase restored to wild-type activity levels (1.58×10(-8)U/CFU). Complementation of P1C5 with MGA_0329 did not restore it to wild-type levels of virulence, indicating that the contribution of sialidase to M. gallisepticum virulence is not straightforward.

  8. The Effect of an Alternate Start Codon on Heterologous Expression of a PhoA Fusion Protein in Mycoplasma gallisepticum.

    PubMed

    Panicker, Indu S; Browning, Glenn F; Markham, Philip F

    2015-01-01

    While the genomes of many Mycoplasma species have been sequenced, there are no collated data on translational start codon usage, and the effects of alternate start codons on gene expression have not been studied. Analysis of the annotated genomes found that ATG was the most prevalent translational start codon among Mycoplasma spp. However in Mycoplasma gallisepticum a GTG start codon is commonly used in the vlhA multigene family, which encodes a highly abundant, phase variable lipoprotein adhesin. Therefore, the effect of this alternate start codon on expression of a reporter PhoA lipoprotein was examined in M. gallisepticum. Mutation of the start codon from ATG to GTG resulted in a 2.5 fold reduction in the level of transcription of the phoA reporter, but the level of PhoA activity in the transformants containing phoA with a GTG start codon was only 63% of that of the transformants with a phoA with an ATG start codon, suggesting that GTG was a more efficient translational initiation codon. The effect of swapping the translational start codon in phoA reporter gene expression was less in M. gallisepticum than has been seen previously in Escherichia coli or Bacillus subtilis, suggesting the process of translational initiation in mycoplasmas may have some significant differences from those used in other bacteria. This is the first study of translational start codon usage in mycoplasmas and the impact of the use of an alternate start codon on expression in these bacteria.

  9. Effects of different vaccine combinations against Mycoplasma gallisepticum on the internal egg and eggshell characteristics of commercial layer chickens

    USDA-ARS?s Scientific Manuscript database

    Live F-strain Mycoplasma gallisepticum (FMG) vaccines are presently being used to help control field strain MG outbreaks. However, they may exert some adverse effects on egg production. Live strains of MG of lesser virulence as well as killed vaccines have little or no effect on egg production, bu...

  10. Effects of live and killed vaccines against Mycoplasma gallisepticum on the performance characteristics of commercial layer chickens

    USDA-ARS?s Scientific Manuscript database

    Mycoplasma gallisepticum (MG) is a major and economically significant pathogen of avian species. Different strains of MG have been used as vaccines in multiple-age commercial layer farms in an effort to protect the birds against more virulent field strains. The lower level of protection afforded b...

  11. A novel transposon construct expressing PhoA with potential for studying protein expression and translocation in Mycoplasma gallisepticum.

    PubMed

    Panicker, Indu S; Kanci, Anna; Chiu, Chien-Ju; Veith, Paul D; Glew, Michelle D; Browning, Glenn F; Markham, Philip F

    2012-07-08

    Mycoplasma gallisepticum is a major poultry pathogen and causes severe economic loss to the poultry industry. In mycoplasmas lipoproteins are abundant on the membrane surface and play a critical role in interactions with the host, but tools for exploring their molecular biology are limited. In this study we examined whether the alkaline phosphatase gene (phoA ) from Escherichia coli could be used as a reporter in mycoplasmas. The promoter region from the gene for elongation factor Tu (ltuf) and the signal and acylation sequences from the vlhA 1.1 gene, both from Mycoplasma gallisepticum , together with the coding region of phoA , were assembled in the transposon-containing plasmid pISM2062.2 (pTAP) to enable expression of alkaline phosphatase (AP) as a recombinant lipoprotein. The transposon was used to transform M. gallisepticum strain S6. As a control, a plasmid containing a similar construct, but lacking the signal and acylation sequences, was also produced (pTP) and also introduced into M. gallisepticum . Using a colorimetric substrate for detection of alkaline phosphatase activity, it was possible to detect transformed M. gallisepticum . The level of transcription of phoA in organisms transformed with pTP was lower than in those transformed with pTAP, and alkaline phosphatase was not detected by immunoblotting or enzymatic assays in pTP transformants, eventhough alkaline phosphatase expression could be readily detected by both assays in pTAP transformants. Alkaline phosphatase was shown to be located in the hydrophobic fraction of transformed mycoplasmas following Triton X-114 partitioning and in the membrane fraction after differential fractionation. Trypsin proteolysis confirmed its surface exposure. The inclusion of the VlhA lipoprotein signal sequence in pTAP enabled translocation of PhoA and acylation of the amino terminal cysteine moiety, as confirmed by the effect of treatment with globomycin and radiolabelling studies with [14C]palmitate. PhoA could be

  12. A novel transposon construct expressing PhoA with potential for studying protein expression and translocation in Mycoplasma gallisepticum

    PubMed Central

    2012-01-01

    Background Mycoplasma gallisepticum is a major poultry pathogen and causes severe economic loss to the poultry industry. In mycoplasmas lipoproteins are abundant on the membrane surface and play a critical role in interactions with the host, but tools for exploring their molecular biology are limited. Results In this study we examined whether the alkaline phosphatase gene (phoA ) from Escherichia coli could be used as a reporter in mycoplasmas. The promoter region from the gene for elongation factor Tu (ltuf) and the signal and acylation sequences from the vlhA 1.1 gene, both from Mycoplasma gallisepticum , together with the coding region of phoA , were assembled in the transposon-containing plasmid pISM2062.2 (pTAP) to enable expression of alkaline phosphatase (AP) as a recombinant lipoprotein. The transposon was used to transform M. gallisepticum strain S6. As a control, a plasmid containing a similar construct, but lacking the signal and acylation sequences, was also produced (pTP) and also introduced into M. gallisepticum . Using a colorimetric substrate for detection of alkaline phosphatase activity, it was possible to detect transformed M. gallisepticum . The level of transcription of phoA in organisms transformed with pTP was lower than in those transformed with pTAP, and alkaline phosphatase was not detected by immunoblotting or enzymatic assays in pTP transformants, eventhough alkaline phosphatase expression could be readily detected by both assays in pTAP transformants. Alkaline phosphatase was shown to be located in the hydrophobic fraction of transformed mycoplasmas following Triton X-114 partitioning and in the membrane fraction after differential fractionation. Trypsin proteolysis confirmed its surface exposure. The inclusion of the VlhA lipoprotein signal sequence in pTAP enabled translocation of PhoA and acylation of the amino terminal cysteine moiety, as confirmed by the effect of treatment with globomycin and radiolabelling studies with [14

  13. EFFECTS OF BROILER REARING ENVIRONMENT ON TRANSMISSION OF F-STRAIN MYCOPLASMA GALLISEPTICUM FROM COMMERCIAL LAYER HENS TO BROILER CHICKENS: ROLE OF ACID-BASE BALANCE

    USDA-ARS?s Scientific Manuscript database

    Two trials were conducted concurrently to determine and compare, blood pH, blood gases, hematocrit, and hemoglobin in mycoplasma-free, F-strain Mycoplasma gallisepticum (FMG) inoculation layers, and FMG contact-infected broilers. FMG-inoculated layers had the highest partial pressure of O2 and the l...

  14. Ex vivo pharmacokinetic and pharmacodynamic analysis of valnemulin against Mycoplasma gallisepticum S6 in Mycoplasma gallisepticum and Escherichia coli co-infected chickens.

    PubMed

    Xiao, Xia; Sun, Jian; Chen, Yi; Zou, Mengting; Zhao, Dong-Hao; Liu, Ya-Hong

    2015-04-01

    Pharmacokinetic and pharmacodynamic (PK/PD) indices against Mycoplasma gallisepticum (MG) S6 were investigated in an ex vivo PK/PD model following oral administration of valnemulin to chickens co-infected with M. gallisepticum and Escherichia coli. The minimum inhibitory concentrations (MICs) for valnemulin against MG S6 in artificial medium and chicken serum were determined. In vitro time-killing curves were established according to a series of multiples of the MIC value in an artificial medium, and ex vivo time-killing curves were established in serum samples obtained from infected chickens at different time points after oral administration with an initial titer of 1 × 10(6) color change units (CCU)/mL MG S6. The sigmoid Emax model was used to provide 24 h area under concentration-time curve/minimum inhibitory concentration ratios (AUC0-24h/MIC) for mycoplasmastasis, mycoplasmacidal activity and mycoplasmal elimination, respectively. The inoculum size and micro or macro methods exhibited little effect on MIC determination of MG, whereas matrix had a large effect. The rapid killing activity observed in in vitro time-killing curves seems to indicate that valnemulin was mycoplasmacidal and concentration dependent against MG. The AUC0-24h/MIC ratio for mycoplasmacidal activity and mycoplasmal elimination was 1321 h and 1960 h, respectively. A dosage regimen of 12.4 mg/kg/day and 18.3 mg/kg/day valnemulin was calculated for mycoplasmacidal activity and mycoplasmal elimination against MG S6, respectively.

  15. Evaluation of two commercial enzyme-linked immunosorbent assay kits for the detection of Mycoplasma gallisepticum antibodies.

    PubMed

    Kempf, I; Gesbert, F; Guittet, M; Bennejean, G; Stipkovits, L

    1994-06-01

    Sensitivity and specificity of two commercial Mycoplasma gallisepticum (MG) enzyme-linked immunosorbent assay (ELISA) kits, rapid slide agglutination (SA) and haemagglutination inhibition (HI) tests were compared using sera from specific pathogen free chickens, turkeys or ducks which had been inoculated with various avian mycoplasmas, bacteria or with a reovirus. Results show that sensitivity of SA was superior to ELISA and HI tests in the ability to detect antibodies formed in early response to MG infection. However, both ELISA kits and HI tests had a higher degree of specificity.

  16. Serologic response of Rio Grande wild turkeys to experimental infections of Mycoplasma gallisepticum

    USGS Publications Warehouse

    Rocke, Tonie E.; Yuill, Thomas M.

    1988-01-01

    The serologic response of Rio Grande wild turkeys (Meleagris gallopavo intermedia) to Mycoplasma gallisepticum (MG) was determined. Free-ranging turkeys were caught in southern Texas, shipped to the University of Wisconsin, Madison, and housed in isolation facilities. Fourteen birds were exposed to MG, by intratracheal and intranasal inoculation. Eight birds received sterile broth only. Two wk prior to the end of the experiment, MG exposed turkeys were stressed by challenge with a serologically unrelated mycoplasma. Serum from all exposed birds reacted positively for MG antibody by the rapid plate agglutination (RPA) procedure within 2 mo postexposure (PE) and all but one remained positive for 14 mo PE. Less than one half of the exposed birds developed positive MG antibody titers detectable by the hemagglutination inhibition (HI) test within 2 mo PE, and by 10 mo PE, none had positive titers. Antibody was detected by the HI test in two of 11 infected turkeys, 14 mo PE, and titers increased significantly within 2 wk. MG was isolated from tracheal swabs from two infected birds 2 mo PE, but attempts thereafter failed. However, at the termination of the experiment 15 mo later, MG was isolated from lung tissue of three of 11 exposed turkeys and from a blood clot found in the lower trachea of one bird.

  17. The effect of the air sampling method on the recovery of Mycoplasma gallisepticum from experimentally produced aerosols.

    PubMed

    Landman, W J M; Feberwee, A; van Eck, J H H

    2013-06-01

    A reliable air sampling method is a prerequisite to calculate the inhaled aerosol dose by animals exposed to the aerosol as precise as possible.[Comp]: Set abstract according to the journal style.[/Comp] To examine if aerosol collection in a fluid medium (buffered peptone water (BPW) in the impinger) improves detection of viable mycoplasmas. Also the effect of adding Mycoplasma Experience (ME) broth and/or BPW to the aerosol fluid on aerosol titres was assessed. Aerosols containing a Mycoplasma gallisepticum field or vaccine strain were simultaneously sampled with gelatin filters and by impinger immediately after ending aerosolization and 25 min later. Sampling of M. gallisepticum aerosols using the impinger did not yield higher aerosol titres compared to sampling with gelatin filters. Initial loss during generation of the field strain aerosol and the half-life time of viable mycoplasmas in the aerosol were 1.1-2.4 log10 and <4-15 min, respectively. The vaccine strain was more vulnerable compared to its field counterpart. In spite of higher aerosolized doses of the vaccine strain (10(8.0) to 10(8.1) versus 10(7.5) cfu per m(3) of air of the field strain), mycoplasmas were not recovered from the aerosols neither by gelatin filter nor by impinger. Therefore, half-life times could not be calculated. Addition of BPW to the aerosol fluid did not clearly improve the recovery of the field strain from the aerosol, while addition of ME broth and BPW did. Gelatin filters likely due to their relative high moisture content (10-14% wt/wt) are at least as useful as the impinger for the recovery of M. gallisepticum from aerosols, provided exsiccation of the filters is prevented.

  18. The Effect of an Alternate Start Codon on Heterologous Expression of a PhoA Fusion Protein in Mycoplasma gallisepticum

    PubMed Central

    Panicker, Indu S.; Browning, Glenn F.; Markham, Philip F.

    2015-01-01

    While the genomes of many Mycoplasma species have been sequenced, there are no collated data on translational start codon usage, and the effects of alternate start codons on gene expression have not been studied. Analysis of the annotated genomes found that ATG was the most prevalent translational start codon among Mycoplasma spp. However in Mycoplasma gallisepticum a GTG start codon is commonly used in the vlhA multigene family, which encodes a highly abundant, phase variable lipoprotein adhesin. Therefore, the effect of this alternate start codon on expression of a reporter PhoA lipoprotein was examined in M. gallisepticum. Mutation of the start codon from ATG to GTG resulted in a 2.5 fold reduction in the level of transcription of the phoA reporter, but the level of PhoA activity in the transformants containing phoA with a GTG start codon was only 63% of that of the transformants with a phoA with an ATG start codon, suggesting that GTG was a more efficient translational initiation codon. The effect of swapping the translational start codon in phoA reporter gene expression was less in M. gallisepticum than has been seen previously in Escherichia coli or Bacillus subtilis, suggesting the process of translational initiation in mycoplasmas may have some significant differences from those used in other bacteria. This is the first study of translational start codon usage in mycoplasmas and the impact of the use of an alternate start codon on expression in these bacteria. PMID:26010086

  19. Evaluation of the egg transmission and pathogenicity of Mycoplasma gallisepticum isolates genotyped as ts-11.

    PubMed

    Armour, Natalie K; Ferguson-Noel, Naola

    2015-01-01

    Live Mycoplasma gallisepticum vaccines are used for the control of respiratory disease, egg production losses and egg transmission associated with M. gallisepticum infection in long-lived poultry. The first field case of apparent increased virulence and vertical transmission of ts-11, a live M. gallisepticum vaccine, has been reported. In that study a M. gallisepticum isolate from the broiler progeny of ts-11-vaccinated breeders was genotyped as ts-11 by sequence analysis of four different genetic targets and Random Amplified Polymorphic DNA and found to be significantly more virulent than ts-11 vaccine. The objective of the current study was to evaluate the rate of egg transmission and pathogenicity of ts-11 vaccine and isolates recovered from ts-11-vaccinated breeders (K6222B) and their broiler progeny (K6216D) which had been genotyped as ts-11. Groups of 28-week-old specific pathogen-free chickens at 87% average weekly egg production were inoculated with sterile broth media (negative controls), ts-11 vaccine, K6222B, K6216D or R strain (positive controls) by eye-drop and aerosol. K6216D transmitted via the egg at an average rate of 4.0% in the third and fourth weeks post-infection, while egg transmission of K6222B and ts-11 vaccine was not detected. M. gallisepticum was isolated from the air sacs, ovaries and oviducts of hens infected with K6216D and K6222B, but not from those infected with ts-11 vaccine. K6216D and K6222B both induced respiratory signs and significantly more tracheal colonization and more severe tracheal and air sac lesions than ts-11 vaccine (P ≤ 0.05). There were no substantial differences in the egg production of ts-11, K6216D and K6222B infected groups. These results provide the first conclusive evidence of transovarian transmission of an isolate genotyped as ts-11 and indicate that isolates genotyed as ts-11 vary in their virulence and ability to transmit via the egg.

  20. Effects of different vaccine combinations against Mycoplasma gallisepticum on blood characteristics in commercial layer chickens.

    PubMed

    Peebles, E David; Jacob, Roymon; Branton, Scott L; Evans, Jeffrey D; Leigh, Spencer A; Gerard, Patrick D

    2015-09-01

    Mycoplasma gallisepticum (MG) is a major and economically significant pathogen of avian species. When administered before lay, F-strain MG (FMG) can reduce egg production during lay, but the ts-11 strain of MG (ts11MG) does not exert this effect. Two trials were conducted to determine the effects of pre-lay vaccinations of ts11MG, MG-Bacterin (MGBac), or their combination, in conjunction with an FMG challenge overlay after peak production on the blood characteristics of commercial layers. In each trial, 160 mycoplasma-free Hy-Line W-36 layers were housed in negative-pressure biological isolation units (4 units per treatment, 10 birds per unit) from 9 through 52 wk of age (woa). The following vaccination treatments were administered at 10 woa: 1) Control (no vaccinations); 2) MGBac; 3) ts11MG; and 4) ts11MG and MGBac combination (ts11MG+MGBac). At 45 woa, half of the birds were challenged with a laboratory stock of high-passage FMG. Parameters measured in both trials were whole-blood hematocrit and serum concentrations of cholesterol (SCHOL), triglycerides, calcium, and total protein (STP). An age×treatment interaction (P=0.04) was observed for STP between 23 and 43 woa. The STP concentration in the ts11MG and ts11MG+MGBac groups was higher at 33 woa, but was lower at 43 woa, in comparison to the Control group. Also, at 38 woa, the STP of the ts11MG+MGBac group was higher than that of the MGBac group. Although use of the ts11MG vaccine alone or in combination with MGBac may influence circulating STP concentrations when administered before lay, it remains effective in protecting layers against the adverse effect of a post-peak challenge of FMG on egg production, as was observed in a previous companion study.

  1. [Antimycoplasmal activities of ofloxacin and commonly used antimicrobial agents on Mycoplasma gallisepticum].

    PubMed

    Takahashi, I; Yoshida, T

    1989-05-01

    In vitro activities of ofloxacin (OFLX), a new quinolone derivative, against 29 strains of Mycoplasma gallisepticum was compared with those of 4 commonly used antimicrobial agents, doxycycline (DOXY), tylosin (TS), spectinomycin (SPCM) and thiamphenicol (TP). Antimycoplasmal activities of the drugs were evaluated on the MIC (final MIC) and MPC (minimum mycoplasmacidal concentration) values which were determined by a broth dilution procedure. The following results were obtained. 1. The MIC90s of OFLX and DOXY were both 0.20 micrograms/ml. The MICs of TS were distributed through a wide range (less than or equal to 0.006 - 0.78 micrograms/ml), and its MIC90 was 0.78 micrograms/ml. Of 29 M. gallisepticum strains, 27.6% were recognized as TS-resistant. The MIC90 values of SPCM and TP were 1.56 micrograms/ml and 3.13 micrograms/ml, respectively. The MIC90 of OFLX was equal to that of DOXY and 4- to 16-fold smaller than the values of the other 3 antibiotics. 2. The MPC of OFLX was the lowest among the antibiotics tested, its MPC90 value was 0.39 micrograms/ml and was followed by DOXY (1.56 micrograms/ml). The MPCs of TS were distributed in a wide range (0.012 - 3.13 micrograms/ml), and its MPC90 was 3.13 micrograms/ml. The MPC90 values of SPCM and TP were both 6.25 micrograms/ml. Therefore, the mycoplasmacidal activity of OFLX evaluated with MPC90 values was 4- to 16-fold greater than those of the other 4 antibiotics.

  2. Mycoplasma gallisepticum and Escherichia coli mixed infection model in broiler chickens for studying valnemulin pharmacokinetics.

    PubMed

    Xiao, X; Zhao, D H; Yang, X; Shi, W; Deng, H; Ma, J; Zhang, S; Liu, Y H

    2014-02-01

    A Mycoplasma gallisepticum-Escherichia coli mixed infection model was developed in broiler chickens, which was applied to pharmacokinetics of valnemulin in the present experiment. The velogenic M. gallisepticum standard strain S6 was rejuvenated to establish the animal model, and the wild E. coli strain O78 was injected as supplementary inoculum to induce chronic respiratory disease in chickens. The disease model was evaluated based on its clinical signs, histopathological examination, bacteriological assay, and serum plate agglutination test. The pharmacokinetics of valnemulin in infected chickens was determined by intramuscular (i.m.) injection and oral administration (per os, p.o.) of a single dose of 10 mg/kg body weight (BW). Plasma samples were analyzed by liquid chromatography-tandem mass spectrometry. The plasma concentration-time curve of valnemulin was analyzed using the noncompartmental method. After the i.m. administration, the mean values of Cmax , Tmax , AUClast , MRT, CLβ /F, Vz /F, and t1⁄2β , were 27.94 μg/mL, 1.57 h, 171.63 μg·h/mL, 4.51 h, 0.06 L/h/kg, 0.56 L/kg, and 6.50 h, respectively. By contrast, the corresponding values after p.o. administration were 5.93 μg/mL, 7.14 h, 47.60 μg·h/mL, 9.80 h, 0.22 L/h/kg, 3.35 L/kg, and 10.60 h. The disposition of valnemulin was retarded in infected chickens after both modes of extravascular administration as compared to the healthy controls. More attention should be given to monitoring the therapeutic efficacy and adverse effects of mixed infection because of higher required plasma drug concentration and enlarged AUC with valnemulin treatment.

  3. The efficacy of Mycoplasma gallisepticum K-strain live vaccine in broiler and layer chickens.

    PubMed

    Ferguson-Noel, N M; Williams, S M

    2015-01-01

    The efficacy of a live Mycoplasma gallisepticum (MG) vaccine candidate (K-strain) was compared to commercially available vaccines in broiler-type chickens (Trial 1) and layer-type chickens (Trial 2). In Trial 1, three-week-old broiler-type chickens were vaccinated via aerosol with K-strain or an F-strain vaccine. The vaccinated chickens and 10 non-vaccinated controls were subsequently challenged with virulent R-strain via aerosol at six weeks post vaccination; both K-strain and F-strain vaccination resulted in significant protection from air sac and tracheal lesions, as well as R-strain colonization (P ≤ 0.05). In Trial 2, commercial layer-type chickens were vaccinated with ts-11 (via eye drop) or K-strain (via aerosol) at 12 weeks of age. At 25 weeks of age these birds were challenged with R-strain via aerosol. The ts-11 and K-strain vaccinated groups both had significantly lower air sac lesion scores and a lower prevalence of ovarian regression after challenge as compared to non-vaccinated chickens (P ≤ 0.05). K-strain vaccination also prevented significant tracheal lesions and R-strain colonization (P ≤ 0.05). K-strain shows great potential as a highly efficacious live MG vaccine in broiler and layer-type chickens for protection of the respiratory and reproductive systems as well as prevention of infection with field strains.

  4. Enhanced conformational flexibility of the histone-like (HU) protein from Mycoplasma gallisepticum.

    PubMed

    Altukhov, Dmitry A; Talyzina, Anna A; Agapova, Yulia K; Vlaskina, Anna V; Korzhenevskiy, Dmitry A; Bocharov, Eduard V; Rakitina, Tatiana V; Timofeev, Vladimir I; Popov, Vladimir O

    2016-12-29

    The histone-like (HU) protein is one of the major nucleoid-associated proteins involved in DNA supercoiling and compaction into bacterial nucleoid as well as in all DNA-dependent transactions. This small positively charged dimeric protein binds DNA in a non-sequence specific manner promoting DNA super-structures. The majority of HU proteins are highly conserved among bacteria; however, HU protein from Mycoplasma gallisepticum (HUMgal) has multiple amino acid substitutions in the most conserved regions, which are believed to contribute to its specificity to DNA targets unusual for canonical HU proteins. In this work, we studied the structural dynamic properties of the HUMgal dimer by NMR spectroscopy and MD simulations. The obtained all-atom model displays compliance with the NMR data and confirms the heterogeneous backbone flexibility of HUMgal. We found that HUMgal, being folded into a dimeric conformation typical for HU proteins, has a labile α-helical body with protruded β-stranded arms forming DNA-binding domain that are highly flexible in the absence of DNA. The amino acid substitutions in conserved regions of the protein are likely to affect the conformational lability of the HUMgal dimer that can be responsible for complex functional behavior of HUMgal in vivo, e.g. facilitating its spatial adaptation to non-canonical DNA-targets.

  5. Role of the GapA and CrmA cytadhesins of Mycoplasma gallisepticum in promoting virulence and host colonization.

    PubMed

    Indiková, Ivana; Much, Peter; Stipkovits, László; Siebert-Gulle, Karin; Szostak, Michael P; Rosengarten, Renate; Citti, Christine

    2013-05-01

    Mycoplasma gallisepticum is an important avian pathogen that commonly induces chronic respiratory disease in chicken. To better understand the mycoplasma factors involved in host colonization, chickens were infected via aerosol with two hemadsorption-negative (HA(-)) mutants, mHAD3 and RCL2, that were derived from a low passage of the pathogenic strain R (Rlow) and are both deficient in the two major cytadhesins GapA and CrmA. After 9 days of infection, chickens were monitored for air sac lesions and for the presence of mycoplasmas in various organs. The data showed that mHAD3, in which the crmA gene has been disrupted, did not promote efficient colonization or significant air sac lesions. In contrast, the spontaneous HA(-) RCL2 mutant, which contains a point mutation in the gapA structural gene, successfully colonized the respiratory tract and displayed an attenuated virulence compared to that of Rlow. It has previously been shown in vitro that the point mutation of RCL2 spontaneously reverts with a high frequency, resulting in on-and-off switching of the HA phenotype. Detailed analyses further revealed that such an event is not responsible for the observed in vivo outcome, since 98.4% of the mycoplasma populations recovered from RCL2-infected chickens still display the mutation and the associated phenotype. Unlike Rlow, however, RCL2 was unable to colonize inner organs. These findings demonstrate the major role played by the GapA and CrmA proteins in M. gallisepticum host colonization and virulence.

  6. Mutations in 23S rRNA gene associated with decreased susceptibility to tiamulin and valnemulin in Mycoplasma gallisepticum.

    PubMed

    Li, Bei-Bei; Shen, Jian-Zhong; Cao, Xing-Yuan; Wang, Yang; Dai, Lei; Huang, Si-Yang; Wu, Cong-Ming

    2010-07-01

    Mycoplasma gallisepticum is a major etiological agent of chronic respiratory disease (CRD) in chickens and sinusitis in turkeys. The pleuromutilin antibiotics tiamulin and valnemulin are currently used in the treatment of M. gallisepticum infection. We studied the in vitro development of pleuromutilin resistance in M. gallisepticum and investigated the molecular mechanisms involved in this process. Pleuromutilin-resistant mutants were selected by serial passages of M. gallisepticum strains PG31 and S6 in broth medium containing subinhibitory concentrations of tiamulin or valnemulin. A portion of the gene encoding 23S rRNA gene (domain V) and the gene encoding ribosome protein L3 were amplified and sequenced. No mutation could be detected in ribosome protein L3. Mutations were found at nucleotide positions 2058, 2059, 2061, 2447 and 2503 of 23S rRNA gene (Escherichia coli numbering). Although a single mutation could cause elevation of tiamulin and valnemulin MICs, combinations of two or three mutations were necessary to produce high-level resistance. All the mutants were cross-resistant to lincomycin, chloramphenicol and florfenicol. Mutants with the A2058G or the A2059G mutation exhibited cross-resistance to macrolide antibiotics erythromycin, tilmicosin and tylosin.

  7. Impact of fowlpox-vectored Mycoplasma gallisepticum vaccine Vectormune FP MG on layer hen egg production and egg quality parameters.

    PubMed

    Leigh, S A; Branton, S L; Evans, J D; Collier, S D

    2013-12-01

    This study was conducted to determine the impact of vaccination with Vectormune FP MG on egg production and egg quality characteristics of Single Comb White Leghorn hens. Due to questions of the efficacy of this vaccine in preventing Mycoplasma gallisepticum-mediated pathology, the ability of this vaccine to protect against postproduction-peak egg losses associated with F-strain M. gallisepticum (FMG) vaccination was also investigated. Vaccination with Vectormune FP MG did not result in any significant change in egg production or egg quality parameters compared with control (unvaccinated) hens. Subsequent revaccination with FMG at 45 wk of age (woa) yielded no impact on egg production or egg quality parameters of Vectormune FP MG vaccinated hens, unlike prior results for postproduction-peak vaccination of M. gallisepticum-clean hens with FMG, which exhibited a drop in egg production of approximately 6%. No difference in egg size distribution was observed for any of the treatment groups before or after FMG revaccination. These results suggest that hens can be safely vaccinated with Vectormune FP MG as pullets and can be revaccinated with a live M. gallisepticum vaccine such as FMG at a later date with no deleterious effects on egg production or egg or eggshell quality parameters.

  8. Effects of different vaccine combinations against Mycoplasma gallisepticum on the digestive and reproductive organ characteristics of commercial egg-laying hens

    USDA-ARS?s Scientific Manuscript database

    Mycoplasma gallisepticum (MG) is a major and economically significant pathogen of avian species. When administered before lay, F-strain MG (FMG) can reduce egg production during lay, but the ts-11 strain of MG (ts11MG) does not exert this effect. Two trials were conducted to determine the effects ...

  9. Serologic response of roosters to gradient dosage levels of a commercially available live F strain-derived Mycoplasma gallisepticum vaccine over time

    USDA-ARS?s Scientific Manuscript database

    Spray application is a commonly used time- and labor-efficient means to deliver live Mycoplasma gallisepticum (MG) vaccine to laying hens in commercial production facilities. The dosage of vaccine received by spray vaccinated birds can vary due to variation in the spray plume and vaccine suspension...

  10. Mycoplasma gallisepticum in the commercial egg-laying hen: an historical perspective considering effects of pathogen strain, age of bird at inoculation, and diet on performance and physiology

    USDA-ARS?s Scientific Manuscript database

    Mycoplasma gallisepticum (MG), a pathogenic organism, primarily causes respiratory distress, but can also spread systemically to subsequently reduce egg production and egg quality in laying hens. However, the effects of MG on the performance and physiology of the commercial laying hen have been sho...

  11. Mycoplasma gallisepticum transmission: Comparison of commercial F-strain vaccine versus layer complex-derived field strains in a tunnel ventilated house

    USDA-ARS?s Scientific Manuscript database

    Two simultaneous trials were conducted using a commercially available, live, F strain Mycoplasma gallisepticum (FMG) vaccine [Trial 1] or two inocula of layer complex-derived MG strains (LCD-MG) [Trial 2]. In each of the two trials, four commercial turkeys were housed in each of two adjoining pens ...

  12. Effects of single and combined Mycoplasma gallisepticums vaccinations on blood electrolytes and acid-base balance in commercial egg-laying hens

    USDA-ARS?s Scientific Manuscript database

    In a previous study, it was shown to occur in response to an F-strain Mycoplasma gallisepticum (FMG) inoculation layers from our laboratory a significant increase in arterial partial pressure of oxygen (pO2), which is generally associated with an oxygen-dependent improvement in tissue oxygenation to...

  13. Transfer of maternal immunoglobulins and antibodies to Mycoplasma gallisepticum and Mycoplasma synoviae to the allantoic and amniotic fluid of chicken embryos.

    PubMed

    Bencina, Dusan; Narat, Mojca; Bidovec, Andrej; Zorman-Rojs, Olga

    2005-12-01

    Maternal antibodies can protect avian embryos against vertically transmitted pathogens during embryogenesis and also young birds after hatching. In contrast to the well-known transfer of maternal immunoglobulin (Ig) G (also termed IgY) from the yolk to embryonic blood, information about the transfer of IgA, IgG and IgM from the egg albumen to the extra-embryonic fluids is very limited. In our study, IgA, IgG and IgM to Mycoplasma gallisepticum and Mycoplasma synoviae were detected in oviduct washings of naturally infected hens and in the corresponding egg albumen samples. In their progeny embryos, IgA, IgG and IgM antibodies to these Mycoplasma species were detected in the allantoic fluid (ALF) and amniotic fluid (AMF) on day 14 of embryonic development (ED). Examination of embryos from chickens immunized with antigens of M. synoviae revealed that the appearance of IgA and IgG and of antibodies to M. synoviae in ALF could vary even among embryos of the same dam. However, IgA, IgG and IgM were detected as early as day 7 of ED in ALF and AMF in certain embryos from hens infected with M. synoviae. In five groups of embryos examined on day 7, IgG to M.synoviae was found in 51% of ALF and 33% of AMF samples. M. synoviae was isolated from 2.3% of ALF samples. IgA to M. synoviae appeared in ALF and AMF on day 12 of ED, and could be found in the majority of AMF samples examined from day 14 onwards. IgM to M. synoviae appeared in AMF on day 13 and in ALF on day 14, but was detected in those fluids less frequently than IgA or IgG.

  14. A Multiplex real-time PCR for detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from Brazilian commercial poultry flocks

    PubMed Central

    Fraga, Aline Padilha; de Vargas, Tatiana; Ikuta, Nilo; Fonseca, André Salvador Kazantzi; Celmer, Álvaro José; Marques, Edmundo Kanan; Lunge, Vagner Ricardo

    2013-01-01

    Mycoplasma gallisepticum (MS) and Mycoplasma synoviae (MS) are important avian pathogens and cause economic losses to the poultry industry. Molecular biology techniques are currently used for a rapid detection of these pathogens and the adoption of control measures of the diseases. The aim of this study was to develop and validate a technique for simultaneous detection of MG and MS by multiplex real time polymerase chain reaction (PCR). The complete assay (Multiplex MGMS) was designed with primers and probes specific for each pathogen and developed to be carried out in a single tube reaction. Vaccines, MG and MS isolates and DNA from other Mycoplasma species were used for the development and validation of the method. Further, 78 pooled clinical samples from different poultry flocks in Brazil were obtained and used to determine the sensitivity and specificity of the technique in comparison to 2 real time PCR assays specific for MG (MG PCR) and MS (MS PCR). The results demonstrated an agreement of 100% (23 positive and 44 negative samples) between Multiplex MGMS and MG PCR in the analysis of 67 samples from MG positive and negative poultry flocks, and an agreement of 96.9% between Multiplex MGMS and MS PCR in the analysis of 64 samples from MS positive and negative poultry flocks. Considering the single amplification tests as the gold standard, the Multiplex MGMS showed 100% of specificity and sensitivity in the MG analysis and 94.7% sensitivity and 100% specificity in the MS analysis. This new assay could be used for rapid analysis of MG and MS in the poultry industry laboratories. PMID:24294247

  15. Infection with Mycoplasma gallisepticum buffers the effects of acute stress on innate immunity in house finches.

    PubMed

    Fratto, Melanie; Ezenwa, Vanessa O; Davis, Andrew K

    2014-01-01

    When wild animals become infected, they still must cope with the rigors of daily life, and, thus, they still can be exposed to acute stressors. The suite of physiological responses to acute stress includes modifying the innate immune system, but infections can also cause similar changes. We examined the effects of an acute stressor (capture stress) on leukocyte abundance and bacteria-killing ability (BKA) in wild birds (house finches Haemorhous mexicanus) with and without a naturally occurring infection (Mycoplasma gallisepticum) to determine whether infection alters the typical immune response to stress. Birds were captured and bled within 3 min (baseline sample) and then held in paper bags for 2 h and bled again (stress sample). From blood smears made at both time points, we obtained estimates of total white blood cell (WBC) counts and relative numbers of each cell. We also measured BKA of plasma at both time points. In uninfected birds (n = 26), total WBC count decreased by 30% over time, while in infected birds (n = 9), it decreased by 6%. Relative numbers of heterophils did not change over time in uninfected birds but increased in infected birds. Combined with a reduction in lymphocyte numbers, this led to a threefold increase in heterophil-lymphocyte values in infected birds after the stressor, compared to a twofold increase in uninfected birds. There was a nonsignificant tendency for BKA to decline with stress in uninfected birds but not in diseased birds. Collectively, these results suggest that infections can buffer the negative effects of acute stress on innate immunity.

  16. Molecular characterization and determination of antimicrobial resistance of Mycoplasma gallisepticum isolated from chickens.

    PubMed

    Pakpinyo, Somsak; Sasipreeyajan, Jiroj

    2007-11-15

    In this study, three consecutive approaches of molecular characterization, determination of minimum inhibitory concentration (MIC) and antimicrobial tested on Mycoplasma gallisepticum (MG) isolated from chicken farms were investigated. These approaches were conducted between 2004 and 2005 to 134 MG samples collected from five different regions of the intensive farming area of Thailand. Twenty MG isolates and four reference strains including S6, F, ts-11, and 6/85 were classified according to Random Amplification of Polymorphic DNA (RAPD) patterns prior to the antimicrobial tests. These isolates exhibited 5 different genotypes (A-E). Consequently, MG isolates representing each genotype were tested on 11 registered antibiotics. The levels of MIC were determined. Three antibiotics, doxycycline (0.20 microg/ml), tiamulin (0.10 microg/ml), and tylosin (0.33 microg/ml), gave the least MICs among all effective drugs. Break point comparisons of each antimicrobial suggested that the MG isolates were most sensitive to lincomycin, oxytetracycline, tiamulin, and tylosin. Some MG isolates had an intermediate effect on josamycin and were resistant to enrofloxacin and erythromycin. Our results also indicated that MG isolated and collected from the region and nearby districts had similar RAPD patterns showing properties of antimicrobial resistance. The RAPD patterns may imply the frequent use of antibiotics and a resistant strain of MG. This is the first report of genetic characterization using RAPD reflected by the levels of MIC against MG. The information is useful to plan for prophylactic and therapeutic impacts on the poultry industry especially in the area of intensive use of antibiotics.

  17. Evaluation of inactivated Mycoplasma gallisepticum oil-emulsion bacterins for protection against airsacculitis in broilers.

    PubMed

    Yoder, H W; Hopkins, S R; Mitchell, B W

    1984-01-01

    Broiler chicks were vaccinated subcutaneously in the neck at various ages with a single 0.5-ml dose of beta-propiolactone-inactivated Mycoplasma gallisepticum (MG) oil-emulsion bacterin. Four weeks later, vaccinated and control chicks were placed in cold environmental cabinets, infected with infectious bronchitis virus intratracheally, and 2 days later challenged by aerosol exposure to live MG broth culture. All chicks were killed 21 days later and scored postmortem for the rate and severity of airsacculitis produced in each group. Broiler chicks vaccinated at 1 day of age had only slight protection against the development of airsacculitis. Results were variable when chicks were vaccinated at 7 days of age, with little evidence of resistance to airsacculitis. However, when broiler chicks were vaccinated with MG bacterins at 11 to 15 days of age, they acquired significant protection against airsacculitis compared with controls. Viable MG organisms were readily isolated from most of the sampled tracheas and air-sac lesions cultured 21 days post-challenge, indicating a lack of protection against infection of the respiratory tract. MG-vaccinated chicks generally produced antibodies readily detectable by the rapid serum-plate test, tube-agglutination, and hemagglutination-inhibition (HI) tests. Some of the vaccinated chicks, but none of the unvaccinated control chicks, developed positive reactions to agar-gel-precipitin tests following challenge. Low HI titers at challenge were not necessarily indicative of lack of protection against the development of airsacculitis, since good protection was often observed in chickens with low to moderate HI titers.

  18. Plasma and tissue pharmacokinetics of marbofloxacin in experimentally infected chickens with Mycoplasma gallisepticum and Escherichia coli.

    PubMed

    Ding, H; Wang, L; Shen, X; Gu, X; Zeng, D; Zeng, Z

    2013-10-01

    The plasma and tissue pharmacokinetics of marbofloxacin in chickens experimentally infected with Mycoplasma gallisepticum and Escherichia coli were studied. Marbofloxacin was given to 66 infected chickens by oral administration at a dosage of 5 mg/kg b.w., once a day for three days. Plasma, brain, kidney, liver, lung, muscle and trachea were collected and marbofloxacin concentrations were analyzed by a high performance liquid chromatography method. In the infected chickens, maximal marbofloxacin concentrations in plasma, brain, kidney, liver, lung, muscle and trachea were 1.84, 1.33, 7.35, 5.61, 3.12, 2.98, and 4.51 g/mL (g); the elimination half-lives of marbofloxacin were 6.8, 2.74, 9.31, 8.45, 9.55, 11.53 and 5.46 h for plasma, brain, kidney, liver, lung, muscle and trachea, respectively. AUC were calculated to be 9.68, 8.04, 45.1, 27.03, 20.56, 19.47, and 32.68 μg/mL (g) for plasma, brain, kidney, liver, lung, muscle and trachea, respectively. Marbofloxacin concentration in tissues except for brain exceeded marbofloxacin concentration in plasma, with AUC(tissue) /AUC(plasma) ranging from 2.01 to 4.66 and Peak(tissue) /Peak(plasma) ranging from 1.62 to 3.99. The results showed that a marbofloxacin dosage of 5 mg/kg administered orally at 24 h intervals may provide successful treatment of chicken with MG and E. coli infection. © 2013 John Wiley & Sons Ltd.

  19. Effect of differing +2 amino acids on export of a heterologous PhoA lipoprotein in Mycoplasma gallisepticum.

    PubMed

    Panicker, Indu S; Kanci, Anna; Markham, Philip F; Browning, Glenn F

    2016-08-01

    The significance of the amino acid adjacent to the amino terminal cysteine of lipoproteins, the +2 amino acid, has been well documented in E. coli and there have also been limited studies on Gram-positive bacteria. In this study we investigated whether there was any preference for specific residues and any targeting role attributable to different residues following the cysteine at the amino terminus in lipoproteins of Mycoplasma gallisepticum. There were found to be distinct preferences in this position that vary considerably from the preferences seen in Gram-positive and Gram-negative bacteria. The effect of different amino acids at the +2 position was studied using the pTAP vector, which has been shown to express PhoA as a lipoprotein. Replacement of the threonine at the +2 position in the PhoA lipoprotein with hydrophobic amino acids resulted in higher levels of expression of alkaline phosphatase, while replacement with hydrophilic amino acids resulted in lower levels of expression of alkaline phosphatase. Changes in the +2 amino acid did not appear to alter export of the PhoA lipoprotein to the membrane fraction, but a difference was seen in susceptibility to proteolysis in PhoA lipoproteins with differing +2 amino acids. This is the first study to examine the role of the +2 amino acid in mycoplasma lipoproteins and establish a difference between M. gallisepticum and Gram-positive and Gram-negative bacteria and will assist in optimization of the design of recombinant lipoprotein genes in mycoplasmas for maximal levels of expression and stability on the cell surface.

  20. Mycoplasma viruses.

    PubMed

    Maniloff, J

    1988-01-01

    Unlike bacterial viruses that infect cells bounded by a cell wall, mycoplasma viruses have evolved to enter and propagate in mycoplasma cells bounded only by a single lipid-protein cell membrane. In addition, mycoplasmas have the smallest amount of genetic information of any known cells, so their complexity is constrained by a limited genetic coding capacity. As a consequence of these host cell differences, mycoplasma viruses have been found to have a variety of structures and replication strategies which are different from those of the bacterial viruses. This article is a critical review of mycoplasma viruses infecting the genera Acholeplasma, Spiroplasma, and Mycoplasma; included are data on classification, morphology and structure, biological and physical properties, chemical composition, and productive and lysogenic replication cycles.

  1. Phase Transition of the Bacterium upon Invasion of a Host Cell as a Mechanism of Adaptation: a Mycoplasma gallisepticum Model

    PubMed Central

    Matyushkina, Daria; Pobeguts, Olga; Butenko, Ivan; Vanyushkina, Anna; Anikanov, Nicolay; Bukato, Olga; Evsyutina, Daria; Bogomazova, Alexandra; Lagarkova, Maria; Semashko, Tatiana; Garanina, Irina; Babenko, Vladislav; Vakhitova, Maria; Ladygina, Valentina; Fisunov, Gleb; Govorun, Vadim

    2016-01-01

    What strategies do bacteria employ for adaptation to their hosts and are these strategies different for varied hosts? To date, many studies on the interaction of the bacterium and its host have been published. However, global changes in the bacterial cell in the process of invasion and persistence, remain poorly understood. In this study, we demonstrated phase transition of the avian pathogen Mycoplasma gallisepticum upon invasion of the various types of eukaryotic cells (human, chicken, and mouse) which was stable during several passages after isolation of intracellular clones and recultivation in a culture medium. It was shown that this phase transition is manifested in changes at the proteomic, genomic and metabolomic levels. Eukaryotic cells induced similar proteome reorganization of M. gallisepticum during infection, despite different origins of the host cell lines. Proteomic changes affected a broad range of processes including metabolism, translation and oxidative stress response. We determined that the activation of glycerol utilization, overproduction of hydrogen peroxide and the upregulation of the SpxA regulatory protein occurred during intracellular infection. We propose SpxA as an important regulator for the adaptation of M. gallisepticum to an intracellular environment. PMID:27775027

  2. Semi-automated Curation of Metabolic Models via Flux Balance Analysis: A Case Study with Mycoplasma gallisepticum

    PubMed Central

    Szczepanek, Steven M.; Johnson, Erik L.; Tulman, Edan R.; Ching, Wei-Mei; Geary, Steven J.; Srivastava, Ranjan

    2013-01-01

    Primarily used for metabolic engineering and synthetic biology, genome-scale metabolic modeling shows tremendous potential as a tool for fundamental research and curation of metabolism. Through a novel integration of flux balance analysis and genetic algorithms, a strategy to curate metabolic networks and facilitate identification of metabolic pathways that may not be directly inferable solely from genome annotation was developed. Specifically, metabolites involved in unknown reactions can be determined, and potentially erroneous pathways can be identified. The procedure developed allows for new fundamental insight into metabolism, as well as acting as a semi-automated curation methodology for genome-scale metabolic modeling. To validate the methodology, a genome-scale metabolic model for the bacterium Mycoplasma gallisepticum was created. Several reactions not predicted by the genome annotation were postulated and validated via the literature. The model predicted an average growth rate of 0.358±0.12, closely matching the experimentally determined growth rate of M. gallisepticum of 0.244±0.03. This work presents a powerful algorithm for facilitating the identification and curation of previously known and new metabolic pathways, as well as presenting the first genome-scale reconstruction of M. gallisepticum. PMID:24039564

  3. 9 CFR 113.408 - Avian mycoplasma antigen.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... ion concentration shall be determined with a pH meter which has been standardized with a pH buffer... with five Mycoplasma gallisepticum antiserums (chicken origin); Mycoplasma Meleagridis Antigen shall be examined for cross-agglutination with five Mycoplasma gallisepticum antiserums (turkey origin) and...

  4. 9 CFR 113.408 - Avian mycoplasma antigen.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... ion concentration shall be determined with a pH meter which has been standardized with a pH buffer... with five Mycoplasma gallisepticum antiserums (chicken origin); Mycoplasma Meleagridis Antigen shall be examined for cross-agglutination with five Mycoplasma gallisepticum antiserums (turkey origin) and...

  5. Effects of live and killed vaccines against Mycoplasma gallisepticum on the performance characteristics of commercial layer chickens.

    PubMed

    Jacob, R; Branton, S L; Evans, J D; Leigh, S A; Peebles, E D

    2014-06-01

    Different vaccine strains of Mycoplasma gallisepticum have been used on multiple-age commercial layer farms in an effort to protect birds against virulent field-strain infections. Use of the F-strain of M. gallisepticum (FMG), as an overlay vaccine during lay, may be necessary because of the lower level of protection afforded by M. gallisepticum vaccines of low virulence given before lay. Two replicate trials were conducted to investigate effects of live and killed M. gallisepticum vaccines administered individually and in combination before lay, in conjunction with an FMG vaccine overlay after peak egg production (EP), on the performance characteristics of commercial layers. The following treatments were utilized at 10 wk of age (woa): 1) control (no vaccinations); 2) ts11 strain M. gallisepticum (ts11MG) vaccine; 3) M. gallisepticum-Bacterin vaccine (MG-Bacterin); and 4) ts11MG and MG-Bacterin vaccines combination. At 45 woa, half of the birds were overlaid with an FMG vaccine. Hen mortality, BW, egg weight, percentage hen-day EP, egg blood spots, and egg meat spots were determined at various time periods between 18 and 52 woa. The data from each trial were pooled. Treatment did not affect performance in interval I (23 to 45 woa). However, during interval II (46 to 52 woa), the EP of control and MG-Bacterin-vaccinated birds that later received an FMG vaccine overlay was lower than that in the other treatment groups. Furthermore, treatment application reduced bird BW during interval II. Despite the effects on BW and EP, no differences were observed for egg blood or meat spots among the various treatments. It is suggested that the vaccination of commercial layers before lay with ts11MG, but not MG-Bacterin, may reduce the negative impacts of an FMG overlay vaccination given during lay. These results establish that the vaccination of pullets with ts11MG in combination with the vaccination of hens with an FMG overlay, for continual protection against field-strain M

  6. gga-miR-101-3p Plays a Key Role in Mycoplasma gallisepticum (HS Strain) Infection of Chicken

    PubMed Central

    Chen, Jiao; Wang, Zaiwei; Bi, Dingren; Hou, Yue; Zhao, Yabo; Sun, Jianjun; Peng, Xiuli

    2015-01-01

    Mycoplasma gallisepticum (MG), one of the most pathogenic Mycoplasma, has caused tremendous economic loss in the poultry industry. Recently, increasing evidence has suggested that micro ribonucleic acids (miRNAs) are involved in microbial pathogenesis. However, little is known about potential roles of miRNAs in MG infection of chicken. In the present study, using miRNA Solexa sequencing we have found that gga-miR-101-3p was up-regulated in the lungs of MG-infected chicken embryos. Moreover, gga-miR-101-3p regulated expression of the host enhancer of zeste homolog 2 (EZH2) through binding to the 3’ un-translated region (3’-UTR) of EZH2 gene. Over-expression of gga-miR-101-3p significantly inhibited EZH2 expression and hence inhibited proliferation of chicken embryonic fibroblast (DF-1 cells) by blocking the G1-to-S phase transition. Similar results were obtained in MG-infected chicken embryos and DF-1 cells, where gga-miR-101-3p was significantly up-regulated, while EZH2 was significantly down-regulated. This study reveals that gga-miR-101-3p plays an important role in MG infection through regulation of EZH2 expression and provides a new insight into the mechanisms of MG pathogenesis. PMID:26633386

  7. Effects of time-specific F-strain Mycoplasma gallisepticum inoculation overlays on prelay ts-11-strain M. gallisepticum vaccination on blood characteristics of commercial laying hens.

    PubMed

    Peebles, E D; Vance, A M; Branton, S L; Collier, S D; Gerard, P D

    2009-05-01

    Two trials were conducted to determine the effects of a prelay ts-11-strain Mycoplasma gallisepticum (ts-11MG) vaccination alone or in combination with subsequent time-specific F-strain M. gallisepticum (FMG) inoculations on the blood characteristics of commercial laying hens. The following 4 treatments were utilized: 1) sham vaccination at 10 wk of age, 2) vaccination of ts-11MG at 10 wk, 3) ts-11MG at 10 wk overlaid by FMG inoculation at 22 wk, and 4) ts-11MG at 10 wk overlaid by FMG at 45 wk. Parameters measured in both trials were whole blood hematocrit, plasma protein, serum cholesterol, serum triglycerides, and serum calcium. No significant age x treatment interactions and no significant age or treatment main effects were observed for any of the blood parameters investigated, except for serum calcium. At wk 22, serum calcium concentrations were increased by vaccination with ts-11MG at 10 wk, and levels were further increased when the ts-11MG vaccination at 10 wk was overlaid by an FMG inoculation at 22 wk. These results suggest that ts-11MG vaccination at 10 wk of age alone or combined with F-strain inoculum overlays at either 22 or 45 wk may be used without any consequential effects on hematocrit or the lipid and protein levels in the blood of commercial layers. Because elevations in serum calcium were not associated with changes in hen performance, as reported in a previous companion article, it is further suggested that prelay ts-11MG vaccination before FMG inoculation overlays during lay may provide adequate protection against field strain M. gallisepticum infections while being innocuous to layer performance.

  8. pMGA Phenotypic Variation in Mycoplasma gallisepticum Occurs In Vivo and Is Mediated by Trinucleotide Repeat Length Variation

    PubMed Central

    Glew, M. D.; Browning, Glenn F.; Markham, Philip F.; Walker, Ian D.

    2000-01-01

    Chickens were infected with a pathogenic strain of Mycoplasma gallisepticum, and the expression of pMGA, the major surface protein, was inferred by examination of colonies from ex vivo cells. Within 2 days postinfection, 40% of cells had ceased the expression of the original pMGA surface protein (pMGA1.1), and by day 6, the majority of recovered cells were in this category. The switch in pMGA phenotype which had occurred in vivo was reversible, since most colonies produced from ex vivo progenitors exhibited frequent pMGA1.1+ sectors. After prolonged in vivo habitation, increasing proportions of recovered cells gave rise to variant pMGA colonies which had switched from the expression of pMGA1.1 to another gene, pMGA1.2, concomitant with the acquisition of a (GAA)12 motif 5′ to its promoter. Collectively, the results suggest that changes in M. gallisepticum pMGA gene expression in vivo are normal, common, and possibly obligate events for successful colonization of the host. Surprisingly, the initial cessation of pMGA1.1 expression occurred in the absence of detectable pMGA antibodies and seemed to precede the adaptive immune response. PMID:10992515

  9. Determination of the Mutant Selection Window and Evaluation of the Killing of Mycoplasma gallisepticum by Danofloxacin, Doxycycline, Tilmicosin, Tylvalosin and Valnemulin.

    PubMed

    Zhang, Nan; Ye, Xiaomei; Wu, Yuzhi; Huang, Zilong; Gu, Xiaoyan; Cai, Qinren; Shen, Xiangguang; Jiang, Hongxia; Ding, Huanzhong

    2017-01-01

    Mycoplasma gallisepticum is a common etiological cause of a chronic respiratory disease in chickens; its increasing antimicrobial resistance compromises the use of tetracyclines, macrolides and quinolones in the farm environment. Mutant selection window (MSW) determination was used to investigate the propensity for future resistance induction by danofloxacin, doxycycline, tilmicosin, tylvalosin and valnemulin. Killing of M. gallisepticum strain S6 by these antimicrobials was also studied by incubating M. gallisepticum into medium containing the compounds at the minimal concentration that inhibits colony formation by 99% (MIC99) and the mutant prevention concentration (MPC). Based on the morphology and colony numbers of M. gallisepticum on agar plates, the four kinds of sera in the order of the applicability for culturing M. gallisepticum were swine serum > horse serum > bovine serum > mixed serum. The MPC/MIC99 values for each agent were as follows: danofloxacin > tilmicosin > tylvalosin > doxycycline > valnemulin. MPC generated more rapid and greater magnitude killing than MIC99 against M. gallisepticum. Under exposure of 105-109 CFU/mL at MPC drug levels, valnemulin had the slowest rate of reduction in viable organisms and danofloxacin had the highest rate of reduction.

  10. Determination of the Mutant Selection Window and Evaluation of the Killing of Mycoplasma gallisepticum by Danofloxacin, Doxycycline, Tilmicosin, Tylvalosin and Valnemulin

    PubMed Central

    Zhang, Nan; Ye, Xiaomei; Wu, Yuzhi; Huang, Zilong; Gu, Xiaoyan; Cai, Qinren; Shen, Xiangguang; Jiang, Hongxia; Ding, Huanzhong

    2017-01-01

    Mycoplasma gallisepticum is a common etiological cause of a chronic respiratory disease in chickens; its increasing antimicrobial resistance compromises the use of tetracyclines, macrolides and quinolones in the farm environment. Mutant selection window (MSW) determination was used to investigate the propensity for future resistance induction by danofloxacin, doxycycline, tilmicosin, tylvalosin and valnemulin. Killing of M. gallisepticum strain S6 by these antimicrobials was also studied by incubating M. gallisepticum into medium containing the compounds at the minimal concentration that inhibits colony formation by 99% (MIC99) and the mutant prevention concentration (MPC). Based on the morphology and colony numbers of M. gallisepticum on agar plates, the four kinds of sera in the order of the applicability for culturing M. gallisepticum were swine serum > horse serum > bovine serum > mixed serum. The MPC/MIC99 values for each agent were as follows: danofloxacin > tilmicosin > tylvalosin > doxycycline > valnemulin. MPC generated more rapid and greater magnitude killing than MIC99 against M. gallisepticum. Under exposure of 105–109 CFU/mL at MPC drug levels, valnemulin had the slowest rate of reduction in viable organisms and danofloxacin had the highest rate of reduction. PMID:28052123

  11. Mycoplasmas: Brain invaders?

    PubMed

    Rosales, Rubén S; Puleio, Roberto; Loria, Guido R; Catania, Salvatore; Nicholas, Robin A J

    2017-08-01

    Mycoplasmas of humans and animals are usually associated with respiratory, autoimmune, genital and joint diseases. Human mycoplasmas have also been known to affect the brain. Severe central nervous system (CNS) diseases, such as encephalitis, have been linked to Mycoplasma pneumoniae and ureaplasma infections. Less well known is the sheep and goat pathogen, Mycoplasma agalactiae, which has been found in large quantities in the brain where it may be responsible for non-purulent encephalitis as well as ataxia in young animals. Experimental intra-mammary infections of sheep with this mycoplasma have resulted in histopathological changes in the CNS. The cattle pathogen, M. bovis, has been reported occasionally in the brains of calves and adult cattle showing a range of histopathological lesions including abscesses and fibrinous meningitis. Two avian pathogens, M. gallisepticum and M. synoviae have been isolated from the brains of poultry showing meningeal vasculitis and encephalitis. There have been no reported detections of two other avian pathogens, M. meleagridis or M. iowae in the CNS. Over the last few decades, mycoplasmas have been isolated from the brains of sea mammals dying in large numbers in the North Sea although it was concluded that their role may be secondary to underlying viral disease. Finally, evidence has been advanced that certain Spiroplasma species may have a role in the development of the transmissible spongiform encephalopathies (TSE). Invasion of the brain by mycoplasmas may be as a result of direct entry following damage to the inner ear as seen with M. bovis or across the blood brain barrier by mechanisms as yet uncertain. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Mycoplasma gallisepticum (HS strain) surface lipoprotein pMGA interacts with host apolipoprotein A-I during infection in chicken.

    PubMed

    Hu, Fuli; Zhao, Chengcheng; Bi, Dingren; Tian, Wei; Chen, Jiao; Sun, Jianjun; Peng, Xiuli

    2016-02-01

    The adhesin protein from Mycoplasma gallisepticum (HS strain), namely pMGA1.2, is required for M. gallisepticum (MG) infection in chicken. However, the host factor(s) that interact with pMGA1.2 is not known. In this study, we prepared the membrane fraction of trachea epithelial cells from chicken embryos. Using an improved virus overlay protein blot assay (VOPBA) and glutathione S-transferase (GST) pull-down assay, we found that pMGA1.2 specifically bound to a ∼30 kDa host protein. This host protein was further identified by mass spectrometry as chicken apolipoprotein A-I (ApoA-I). We expressed and purified the recombinant ApoA-I protein in Escherichia coli and confirmed that it bound to the purified pMGA1.2 protein in vitro. Transiently expressed pMGA1.2 and ApoA-I were colocalized in HeLa cells. Finally, we designed small interfering RNA (siRNA) molecules to knock down the expression of either ApoA-I or pMGA1.2, which inhibited the MG-induced cell cycle disruption in cells of chicken embryo fibroblast cell line (DF-1). Similarly, knockdown of ApoA-I inhibited the cilia loss and damage in chicken trachea cells in MG infection. In summary, ApoA-I may be an essential host factor in MG infection through interacting with pMGA1.2.

  13. Influence of enrofloxacin traces in drinking water to doxycycline tissue pharmacokinetics in healthy and infected by Mycoplasma gallisepticum broiler chickens.

    PubMed

    Gbylik-Sikorska, Malgorzata; Posyniak, Andrzej; Sniegocki, Tomasz; Sell, Bartosz; Gajda, Anna; Sawicka, Anna; Olszewska-Tomczyk, Monika; Bladek, Tomasz; Tomczyk, Grzegorz; Zmudzki, Jan

    2016-04-01

    Most of antibiotics, administrated in the treatment of poultry diseases are dissolved in drinking water, and it can lead to water supply systems contamination, especially when the regular cleaning is not using. This situation can lead to unconscious administration of low doses of antibiotics to untreated animals. The aim of this study was to clarify the impact of the exposure of enrofloxacin traces (500 μg l(-1)) to doxycycline pharmacokinetics in healthy and experimentally Mycoplasma gallisepticum infected broiler chickens., Two experimental groups, received of enrofloxacin in water and all groups, received 20 mg kg(-1) bw of doxycycline. The compounds concentrations in muscles and livers were determined by LC-MS/MS. The maximum drug tissue concentration (Cmax) of doxycycline was highest in liver obtained from infected chickens which, received enrofloxacin traces (ENR + DC/MG). It was about 40% higher than in healthy chickens from group I which received only doxycycline. It was found that the concentration-time curve AUC(0-t) values in group ENR + DC/MG were almost 75% higher than in the group (DC) and 35% higher than in group (ENR + DC) which also received enrofloxacin traces. The constant exposure of broiler chickens on enrofloxacin traces as well as infection, may significantly influenced on doxycycline tissue pharmacokinetic profile. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Identification of genes involved in Mycoplasma gallisepticum biofilm formation using mini-Tn4001-SGM transposon mutagenesis.

    PubMed

    Wang, Yang; Yi, Li; Zhang, Fanqing; Qiu, Xusheng; Tan, Lei; Yu, Shengqing; Cheng, Xiangchao; Ding, Chan

    2017-01-01

    Mycoplasma gallisepticum (MG) is an important pathogen that can cause chronic respiratory disease in chickens and infectious sinusitis in turkeys. MG has the ability to form biofilms. The molecular mechanisms underlying MG biofilm formation are complex and poorly understood. To better understand the mechanisms involved in biofilm formation, mini-Tn4001-SGM, a novel transposon vector containing the gentamicin gene was constructed and electroporated into MG strain Rlow. Of the 738 mutants obtained, 12 had significantly reduced capacity to form biofilms in a polystyrene microtiter-plate biofilm assay. Ten different genes were identified as disrupted in these mutants using genomic walking from the transposon insertion sites and Southern bolt hybridization with a transposon-based probe. Four genes were associated with cellular processes, especially synthesis of extracellular polysaccharide and several lipoproteins encoded. Other genes were associated with translation, metabolism and gene regulation, and one had unknown function. Seven genes identified in this study have been previously associated with biofilm formation in MG or other bacterial species. The other three have not been previously reported to play a role in biofilm formation in MG. In conclusion, a new transposon vector was shown to be a powerful tool for future studies of MG pathogenesis. This study adds to our understanding of the molecular mechanisms involved in MG biofilm formation and may shed light on the persistence of MG infections.

  15. Effects of an S6 strain of Mycoplasma gallisepticum challenge at onset of lay on digestive and reproductive tract characteristics in commercial layers.

    PubMed

    Parker, T A; Branton, S L; Jones, M S; Peebles, E D; Gerard, P D; Willeford, K O; Pharr, G T; Maslin, W R

    2003-01-01

    Mycoplasma gallisepticum (MG), a reproductive/respiratory pathogen in poultry, has been implicated in suboptimum egg production and decreased hatchability. Commercial layer hens raised in a controlled environment were inoculated with the S6 strain of MG at 20 wk of age. The S6 inoculation had no effect on bird weight, egg production, digestive tract weight and length, or histopathologic lesion scores, although significant differences were noted in the lengths and weights of various portions of the reproductive tract. This study shows that S6MG inoculation does not detrimentally affect layer hen performance when in the absence of environmental stressors customary to a caged layer facility.

  16. Influence of supplemental dietary poultry fat, phytase, and 25-hydroxycholecalciferol on the blood characteristics of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum

    USDA-ARS?s Scientific Manuscript database

    The effects of 2 supplemental levels of dietary poultry fat (PF) and the combination of PF, phytase (PHY) and 25-hydroxycholecalciferol [25(OH)D] on the blood characteristics of commercial layers inoculated with F-strain Mycoplasma gallisepticum (FMG) were investigated in 2 trials. Sham and FMG ino...

  17. Influence of supplemental dietary poultry fat, phytase, and 25-hydroxycholecalciferol on the egg characteristics of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum

    USDA-ARS?s Scientific Manuscript database

    The effects of 2 supplemental levels of dietary poultry fat (PF) and the combination of PF, phytase (PHY) and 25-hydroxycholecalciferol (D3) on the egg characteristics of commercial layers inoculated with F-strain Mycoplasma gallisepticum (FMG) were investigated in 2 trials. Sham and FMG inoculatio...

  18. Influence of Supplemental Dietary Poultry Fat, Phytase, and 25-Hydroxycholecalciferol on the Performance of Commercial Layers Inoculated Before or at the Onset of Lay with F-Strain Mycoplasma gallisepticum

    USDA-ARS?s Scientific Manuscript database

    The effects of 2 levels of supplemental dietary poultry fat (PF) and the combination of PF, phytase (PHY) and 25-hydroxycholecalciferol (D3) on the performance of commercial layers inoculated with F-strain Mycoplasma gallisepticum (FMG) were investigated in 2 trials. Sham and FMG inoculations were ...

  19. Dietary poultry fat, phytase, and 25-hydroxycholecalciferol influence the digestive and reproductive organ characteristic of commercial...at the onset of lay with F-strain Mycoplasma gallisepticum 1 , 2

    USDA-ARS?s Scientific Manuscript database

    ABSTRACT Effects of 2 supplemental concentrations of dietary poultry fat (PF) and the combination of PF, phytase (PHY) and 25-hydroxycholecalciferol [25(OH)D] on the gross digestive and reproductive organ characteristics of commercial layers inoculated with F-strain Mycoplasma gallisepticum (FMG) w...

  20. Effects of Supplemental Dietary Phytase and 25-Hydroxycholecalciferol on the Performance Characteristics of Commercial Layers Inoculated before or at the Onset of Lay with the F-Strain of Mycoplasma gallisepticum

    USDA-ARS?s Scientific Manuscript database

    The effects of dietary supplementation with phytase and 25-hydroxycholecalciferol on the performance characteristics of commercial layers that were inoculated prelay (12 wk of age) or at the onset of lay (22 wk of age) with F-strain Mycoplasma gallisepticum were assessed. Experimental layer diets, w...

  1. Characterization of in vivo-acquired resistance to macrolides of Mycoplasma gallisepticum strains isolated from poultry

    PubMed Central

    2011-01-01

    The macrolide class of antibiotics, including tylosin and tilmicosin, is widely used in the veterinary field for prophylaxis and treatment of mycoplasmosis. In vitro susceptibility testing of 50 strains of M. gallisepticum isolated in Israel during the period 1997-2010 revealed that acquired resistance to tylosin as well as to tilmicosin was present in 50% of them. Moreover, 72% (13/18) of the strains isolated from clinical samples since 2006 showed acquired resistance to enrofloxacin, tylosin and tilmicosin. Molecular typing of the field isolates, performed by gene-target sequencing (GTS), detected 13 molecular types (I-XIII). Type II was the predominant type prior to 2006 whereas type X, first detected in 2008, is currently prevalent. All ten type X strains were resistant to both fluoroquinolones and macrolides, suggesting selective pressure leading to clonal dissemination of resistance. However, this was not a unique event since resistant strains with other GTS molecular types were also found. Concurrently, the molecular basis for macrolide resistance in M. gallisepticum was identified. Our results revealed a clear-cut correlation between single point mutations A2058G or A2059G in domain V of the gene encoding 23S rRNA (rrnA, MGA_01) and acquired macrolide resistance in M. gallisepticum. Indeed, all isolates with MIC ≥ 0.63 μg/mL to tylosin and with MIC ≥ 1.25 μg/mL to tilmicosin possess one of these mutations, suggesting an essential role in decreased susceptibility of M. gallisepticum to 16-membered macrolides. PMID:21810258

  2. Characterization of in vivo-acquired resistance to macrolides of Mycoplasma gallisepticum strains isolated from poultry.

    PubMed

    Gerchman, Irena; Levisohn, Sharon; Mikula, Inna; Manso-Silván, Lucía; Lysnyansky, Inna

    2011-08-02

    The macrolide class of antibiotics, including tylosin and tilmicosin, is widely used in the veterinary field for prophylaxis and treatment of mycoplasmosis. In vitro susceptibility testing of 50 strains of M. gallisepticum isolated in Israel during the period 1997-2010 revealed that acquired resistance to tylosin as well as to tilmicosin was present in 50% of them. Moreover, 72% (13/18) of the strains isolated from clinical samples since 2006 showed acquired resistance to enrofloxacin, tylosin and tilmicosin. Molecular typing of the field isolates, performed by gene-target sequencing (GTS), detected 13 molecular types (I-XIII). Type II was the predominant type prior to 2006 whereas type X, first detected in 2008, is currently prevalent. All ten type X strains were resistant to both fluoroquinolones and macrolides, suggesting selective pressure leading to clonal dissemination of resistance. However, this was not a unique event since resistant strains with other GTS molecular types were also found. Concurrently, the molecular basis for macrolide resistance in M. gallisepticum was identified. Our results revealed a clear-cut correlation between single point mutations A2058G or A2059G in domain V of the gene encoding 23S rRNA (rrnA, MGA_01) and acquired macrolide resistance in M. gallisepticum. Indeed, all isolates with MIC ≥ 0.63 μg/mL to tylosin and with MIC ≥ 1.25 μg/mL to tilmicosin possess one of these mutations, suggesting an essential role in decreased susceptibility of M. gallisepticum to 16-membered macrolides.

  3. Mechanism of action of the copper(I) complex of 2,9-dimethyl-1,10-phenanthroline on Mycoplasma gallisepticum

    SciTech Connect

    Smit, H.; van der Goot, H.; Nauta, W.T.; Timmerman, H.; de Bolster, M.W.; Stouthamer, A.H.; Vis, R.D.

    1982-06-01

    Evidence was found that the inhibitory action of Cu(DMP)/sub 2/NO/sub 3/, the copper(I) complex of 2,9-dimethyl-1,10-phenanthroline (DMP), on Mycoplasma gallisepticum is a consequence of the ultimate toxicity of copper, and not that of the ligand, DMP. From uptake studies with radiolabeled /sup 67/Cu and (/sup 14/C)DMP, we concluded that significantly more copper than DMP is bound to the mycoplasmal cell. It appeared that dissociation of Cu(DMP)2+ occurred shortly after interaction with the cell membrane. Copper was transported across the cytoplasmic membrane. A strong dependence of copper uptake on the incubation medium was observed in the absence of DMP. The main function of the ligand DMP appeared to be as a vehicle for the transport of copper from nontoxic copper-medium complexes to membrane-buried cellular ligands.

  4. The macrophage inflammatory protein-1beta in the supernatants of Mycoplasma gallisepticum-infected chicken leukocytes attracts the migration of chicken heterophils and lymphocytes.

    PubMed

    Lam, K M

    2002-01-01

    Chicken monocytes, macrophages, heterophils and thrombocytes were infected with Mycoplasma gallisepticum, and their supernatants were collected and tested for the presence of chemotactic activities. The supernatants from MG-infected monocytes and macrophages were able to attract the migration of both heterophils and lymphocytes. The chemotactic activity in these supernatants could be abolished by antibodies prepared against the 10 amino acid peptides of the macrophage inflammatory protein (MIP)-1beta, indicating that the released chemoattractant was a MIP-1beta-like compound. The supernatant from MG-infected heterophils was also able to attract the migration of chicken lymphocytes, but its activity could not be neutralized by the antibody to MIP-1beta, indicating that the chemoattractant is not related to MIP-1beta. The supernatants from both control and MG-infected thrombocytes were able to attract the migration of lymphocytes. These results indicate that there is more than one chemotactic factor that is released by these cells; one of the chemoattractants has been identified as a MIP-1beta. These results also show that MIP-1beta may play a role in the recruitment and accumulation of heterophils and lymphocytes to the sites of mycoplasma infection.

  5. [Pathogenic factors of mycoplasma].

    PubMed

    Shimizu, Takashi

    2015-01-01

    Mycoplasmas are smallest organisms capable of self-replication and cause various diseases in human. Especially, Mycoplasma pneumoniae is known as an etiological agent of pneumonia. From 2010 to 2012, epidemics of M. pneumoniae infections were reported worldwide (e.g., in France, Israel, and Japan). In the diseases caused by mycoplasmas, strong inflammatory responses induced by mycoplasmas have been thought to be important. However, mycoplasmas lack of cell wall and do not possess inflammation-inducing endotoxin such as lipopolysaccharide (LPS). We purified inflammation-inducing factors from pathogenic mycoplasmas and identified that they were lipoproteins. Lipoproteins derived from mycoplasmas induced inflammatory responses through Toll-like receptor (TLR) 2. In addition, we demonstrated that cytadherent property of M. pneumoniae played an important role in induction of inflammatory responses. Cytadherent property of M. pneumoniae induced inflammatory responses through TLR2 independent pathway. TLR4, inflammasomes, and autophagy were involved in this TLR2 independent induction of inflammatory responses.

  6. Feline hemotropic mycoplasmas.

    PubMed

    Sykes, Jane E

    2010-11-01

    Three species of hemotropic mycoplasmas are known to infect cats worldwide, Mycoplasma haemofelis, "Candidatus Mycoplasma turicensis" and "Candidatus Mycoplasma haemominutum." These organisms were previously known as Haemobartonella felis, but are now known to be mycoplasmas. Assays based on polymerase chain reaction technology are the most sensitive and specific diagnostic tests available for these organisms. M haemofelis is the most pathogenic species, and causes hemolytic anemia in immunocompetent cats. Other differential diagnoses for hemolytic anemia should be considered in cats testing positive for "Candidatus Mycoplasma turicensis" and "Candidatus Mycoplasma haemominutum," because the presence of these organisms is not always associated with anemia. Blood from infected cats should be handled with care because of the potential zoonotic nature of hemoplasma infections. The treatment of choice for cats with clinical disease is doxycycline.

  7. RESPIRATORY PATHWAYS IN THE MYCOPLASMA I.

    PubMed Central

    Smith, S. L.; Van Demark, P. J.; Fabricant, J.

    1963-01-01

    Smith, S. L. (Cornell University, Ithaca, N.Y.), P. J. Van Demark, and J. Fabricant. Respiratory pathways in the Mycoplasma. I. Lactate oxidation by Mycoplasma gallisepticum. J. Bacteriol. 86:893–897. 1963.—Resting cells of Mycoplasma gallisepticum 293 required the addition of nicotinamide adenine dinucleotide, thiamine pyrophosphate, and flavine mononucleotide for the maximal rate of sodium lactate oxidation. Inhibitor studies, as well as spectrophotometric and chemical assays, indicate that the pathway of electron transport to oxygen during lactate oxidation does not involve heme catalysts, and is mediated by flavin-linked enzyme systems. The presence of reduced nicotinamide adenine dinucleotide-specific lactic dehydrogenase, menadione reductase, ferricyanide reductase, and reduced nicotinamide adenine dinucleotide oxidase activities was detected in cell-free extracts. No cytochrome c reductase or reduced nicotinamide adenine dinucleotide peroxidase activity was detected in these extracts. PMID:14080798

  8. Effects of a Prelay 6/85-strain Mycoplasma gallisepticum Inoculation Alone or in Conjunction with Subsepuent F-Strain Mycoplasma gallisepticum Inoculation During Lay on the Internal Egg Characteristics of .....

    USDA-ARS?s Scientific Manuscript database

    The effects of a pre-lay 6/85-strain M. gallisepticum (6/85MG) inoculation alone or in conjunction with F-strain M. gallisepticum (FMG) inoculation overlays during lay on the internal egg characteristics of commercial egg laying hens were investigated. In the first 2 treatment groups, birds were sh...

  9. Seasonality and wildlife disease: how seasonal birth, aggregation and variation in immunity affect the dynamics of Mycoplasma gallisepticum in house finches.

    PubMed Central

    Hosseini, Parviez R.; Dhondt, André A.; Dobson, Andy

    2004-01-01

    We examine the role of host seasonal breeding, host seasonal social aggregation and partial immunity in affecting wildlife disease dynamics, focusing on the dynamics of house finch conjunctivitis (Mycoplasma gallisepticum (MG) in Carpodacus mexicanus). This case study of an unmanaged emerging infectious disease provides useful insight into the important role of seasonal factors in driving ongoing disease dynamics. Seasonal breeding can force recurrent epidemics through the input of fresh susceptibles, which will clearly affect a wide variety of wildlife disease dynamics. Seasonal patterns of social aggregation and foraging behaviour could change transmission dynamics. We use latitudinal variation in the timing of breeding, and social systems to model seasonal dynamics of house finch conjunctivitis across eastern North America. We quantify the patterns of seasonal breeding, and social aggregation across a latitudinal gradient in the eastern range of the house finch, supplemented with known field and laboratory information on immunity to MG in finches. We then examine the interactions of these factors in a theoretical model of disease dynamics. We find that both forms of seasonality could explain the dynamics of the house finch-MG system, and that these factors could have important effects on the dynamics of wildlife diseases generally. In particular, while either alone is sufficient to create recurrent cycles of prevalence in a population with an endemic disease, both are required to produce the specific semi-annual pattern of disease prevalence seen in the house finch conjunctivitis system. PMID:15615682

  10. Mycoplasma gallisepticum Lipid Associated Membrane Proteins Up-regulate Inflammatory Genes in Chicken Tracheal Epithelial Cells via TLR-2 Ligation through an NF-κB Dependent Pathway

    PubMed Central

    Majumder, Sanjukta; Zappulla, Frank; Silbart, Lawrence K.

    2014-01-01

    Mycoplasma gallisepticum-mediated respiratory inflammation in chickens is associated with accumulation of leukocytes in the tracheal submucosa. However the molecular mechanisms underpinning these changes have not been well described. We hypothesized that the initial inflammatory events are initiated upon ligation of mycoplasma lipid associated membrane proteins (LAMP) to TLRs expressed on chicken tracheal epithelial cells (TEC). To test this hypothesis, live bacteria or LAMPs isolated from a virulent (Rlow) or a non-virulent (Rhigh) strain were incubated with primary TECs or chicken tracheae ex vivo. Microarray analysis identified up-regulation of several inflammatory and chemokine genes in TECs as early as 1.5 hours post-exposure. Kinetic analysis using RT-qPCR identified the peak of expression for most genes to be at either 1.5 or 6 hours. Ex-vivo exposure also showed up-regulation of inflammatory genes in epithelial cells by 1.5 hours. Among the commonly up-regulated genes were IL-1β, IL-6, IL-8, IL-12p40, CCL-20, and NOS-2, all of which are important immune-modulators and/or chemo-attractants of leukocytes. While these inflammatory genes were up-regulated in all four treatment groups, Rlow exposed epithelial cells both in vitro and ex vivo showed the most dramatic up-regulation, inducing over 100 unique genes by 5-fold or more in TECs. Upon addition of a TLR-2 inhibitor, LAMP-mediated gene expression of IL-1β and CCL-20 was reduced by almost 5-fold while expression of IL-12p40, IL-6, IL-8 and NOS-2 mRNA was reduced by about 2–3 fold. Conversely, an NF-κB inhibitor abrogated the response entirely for all six genes. miRNA-146a, a negative regulator of TLR-2 signaling, was up-regulated in TECs in response to either Rlow or Rhigh exposure. Taken together we conclude that LAMPs isolated from both Rhigh and Rlow induced rapid, TLR-2 dependent but transient up-regulation of inflammatory genes in primary TECs through an NF-κB dependent pathway. PMID:25401327

  11. Chicken gga-miR-19a Targets ZMYND11 and Plays an Important Role in Host Defense against Mycoplasma gallisepticum (HS Strain) Infection

    PubMed Central

    Hu, Qingchang; Zhao, Yabo; Wang, Zaiwei; Hou, Yue; Bi, Dingren; Sun, Jianjun; Peng, Xiuli

    2016-01-01

    Mycoplasma gallisepticum (MG), one of the most pathogenic Mycoplasmas, can cause chronic respiratory disease (CRD) in chickens. It has been suggested that micro-ribonucleic acids (miRNAs) are involved in microbial pathogenesis. However, little is known about the roles of miRNAs in MG infection. Previously, we found by deep sequencing that gga-miR-19a was significantly up-regulated in the lungs of MG-infected chicken embryos. In this work, we confirmed that gga-miR-19a was up-regulated in both MG-infected chicken embryonic lungs and MG-infected DF-1 (chicken embryo fibroblast) cells. At 72 h post-transfection, we found that the over-expression of gga-miR-19a significantly enhanced the proliferation of MG-infected DF-1 cells by promoting the transition from the G1 phase to the S and G2 phases, while a gga-miR-19a inhibitor repressed the proliferation of MG-infected DF-1 cells by arresting the cell cycle in the G1 phase. Moreover, we found that gga-miR-19a regulated the expression of the host zinc-finger protein, MYND-type containing 11 (ZMYND11), through binding to its 3′ untranslated region (3′-UTR). DAVID analysis revealed that ZMYND11 could negatively regulate the NF-kappaB (NF-κB) signaling pathway in chickens (Gallus gallus). Upon MG infection, gga-miR-19a, NF-κB, MyD88, and TNF-α were all up-regulated, whereas ZMYND11 was down-regulated. The over-expression of gga-miR-19a in the DF-1 cells did not affect the above gene expression patterns, and gga-miR-19a inhibitor repressed the expression of NF-κB, MyD88, and TNF-α, but enhanced the expression of ZMYND11. In conclusion, gga-miR-19a might suppress the expression of ZMYND11 in MG-infected chicken embryonic lungs and DF-1 cells, activate the NF-κB signaling pathway, and promote pro-inflammatory cytokines expression, the cell cycle progression and cell proliferation to defend against MG infection. PMID:27683641

  12. Effects of different vaccine combinations against Mycoplasma gallisepticum on the digestive and reproductive organ characteristics of commercial egg-laying hens.

    PubMed

    Peebles, E D; Jacob, R; Branton, S L; Evans, J D; Leigh, S A; Gerard, P D

    2015-12-01

    Mycoplasma gallisepticum (MG) is a major and economically significant pathogen of avian species. When administered before lay, F-strain MG (FMG) can reduce egg production during lay, but the ts-11 strain of MG (ts11MG) does not exert this effect. Two trials were conducted to determine the effects of pre-lay vaccinations of ts11MG, MG-Bacterin (MGBac), or their combination, in conjunction with an FMG vaccination overlay after peak production on the digestive and reproductive organ characteristics of Hy-Line W-36 layers housed in biological isolation units (4 units per treatment, 10 birds per unit). The following vaccination treatments were administered at 10 wk of age (woa): 1) Control (no vaccinations); 2) MGBac; 3) ts11MG; and 4) ts11MG and MGBac combination (ts11MG+MGBac). At 45 woa, half of the birds were vaccinated with a laboratory stock of high passage FMG. In both trials, parameters determined in 4 birds per unit at 55 woa included: BW; fatty liver hemorrhagic syndrome incidence; mean number of mature ovarian follicles; ovarian, oviduct, and small intestine weights; and the weights and lengths of the various portions of the oviduct and small intestine. Treatment effects were observed for the weights of the entire small intestine and the duodenum, jejunum, and ileum, as percentages of BW; and for vagina weight as a percentage of total oviduct weight. In general, the weights of the small intestine and its 3 components were increased in response to the FMG vaccine that was administered at 45 woa. An FMG vaccination at 45 woa may increase relative intestine weight in layers; however, use of a prelay MGBac vaccine alone or in combination with ts11MG, with or without an FMG overlay, does not affect the gross characteristics of their digestive and reproductive organs, and may be used without having an adverse effect on their performance, as was observed in a previous companion study.

  13. Evaluation of the Capacity of PCR and High-Resolution Melt Curve Analysis for Identification of Mixed Infection with Mycoplasma gallisepticum Strains.

    PubMed

    Ghorashi, Seyed A; Kanci, Anna; Noormohammadi, Amir H

    2015-01-01

    Pathogenicity and presentation of Mycoplasma gallisepticum (MG) infection may differ from one strain to another and this may have implications on control measures. Infection of individual birds with more than one MG strain has been reported. A PCR followed by high resolution melt (HRM) curve analysis has been developed in our laboratory and routinely used for detection and differentiation of MG strains. However the potential of this test for identification of MG strains in a mixed specimen has not been evaluated. In the present study, the capability of PCR-HRM curve analysis technique, targeting vlhA and pvpA genes was assessed for identification of individual MG strains in a mixed population. Different DNA ratios of two MG strains from 1 to 10(-4) ng were tested with some generated conventional and normalized curves distinct from those of individual strains alone. Using genotype confidence percentages (GCP) generated from HRM curve analysis, it was found that vlhA PCR-HRM was more consistent than pvpA PCR-HRM for the detection of MG ts-11 vaccine strain mixed with any of the MG strains 6/85, F, S6 or a field isolate. The potential of vlhA PCR-HRM to detect mixed MG strains in a specimen was found to be primarily dependent on quantity and proportion of the target DNAs in the mixture. This is the first study examining the capacity of PCR-HRM technique for identification of individual MG strains in a mixed strain population.

  14. Effects of F-strain Mycoplasma gallisepticum inoculation at twelve weeks of age on digestive and reproductive organ characteristics of commercial egg laying hens.

    PubMed

    Burnham, M R; Peebles, E D; Branton, S L; Jones, M S; Gerard, P D; Maslin, W R

    2002-12-01

    Experimental inoculation with the F-strain of Mycoplasma gallisepticum (FMG) between 8 and 18 wk of age is known to affect reproductive performance in commercial layers. Therefore, two trials were conducted to determine if changes in digestive and reproductive organ characteristics also occur in commercial laying hens infected with FMG at 12 wk of age. In Trial 1, liver weight, liver lipid and moisture contents, ovary weight, ovarian follicular hierarchy, and the weights, lengths, and histologies of the infundibulum, magnum, isthmus, uterus, and vagina were determined. In Trial 2, fatty liver hemorrhagic syndrome (FLHS) incidence and the weights, lengths, and histologies of the duodenum, jejunum, and ileum were determined in addition to the parameters examined in Trial 1. In both trials, the average number of mature (diameter > or = 12 mm) ovarian follicles was lower in FMG-inoculated hens in comparison to controls. Also, magnum/oviduct (cm/cm) length was reduced in treated birds. In Trial 2, isthmus/BW and isthmus/oviduct (g/ g) weight were decreased at 46 wk of age, and vagina/ BW and vagina/oviduct (g/g) weight were decreased at both 20 and 36 wk of age due to FMG treatment. In Trial 2, FMG treatment resulted in a 50% increase in the number of FLHS birds. Furthermore, treatment caused a decrease at 20 wk of age and an increase at 44 wk of age in liver moisture content. However, the intestinal characteristics examined were not affected by FMG inoculation. Altered liver, ovarian, and reproductive organ characteristics were associated with FMG infection in commercial layers. More specifically, FMG inoculation at 12 wk resulted in a higher incidence of FLHS, ovarian follicular regression, and decreased isthmal and vaginal proportions of the reproductive tract. These data clearly demonstrate that alterations in performance and egg characteristics of layers inoculated with FMG at 12 wk of age are related to mutual functional disturbances in the liver, ovary, and

  15. Development and immunogenicity of recombinant GapA(+) Mycoplasma gallisepticum vaccine strain ts-11 expressing infectious bronchitis virus-S1 glycoprotein and chicken interleukin-6.

    PubMed

    Shil, Pollob K; Kanci, Anna; Browning, Glenn F; Markham, Philip F

    2011-04-12

    Mycoplasma gallisepticum (MG) is a major pathogen of poultry that causes chronic respiratory disease in chickens and infectious sinusitis in turkeys. A live attenuated vaccine, ts-11, has been used for the control of MG in several countries. The efficacy of this vaccine is highly dose dependent and the flock antibody response is weak. To improve the functionality of the vaccine and investigate its potential as a delivery vector for foreign antigens and immunomodulatory proteins, we developed a derivative of ts-11 expressing infectious bronchitis virus-S1 glycoprotein (IBV-S1) and releasing chicken interleukin-6 into the extracellular milieu (MG ts-11 C3 (+CS)) using a transposon-based delivery vector. Following administration of MG ts-11 C3 (+CS) to chickens by eye-drop, an antibody response to MG and IBV-S1, as determined by the rapid serum agglutination test (RSA) and Western blotting, respectively, could be detected. Birds inoculated with the recombinant vaccine had significantly enhanced weight gain and were partially protected against damage by pathogenic IBV. These results indicate that the ChIL-6 released by MG ts-11 C3 (+CS) may have had a non-specific effect on growth rate. They also suggest that ts-11 is a promising vaccine vector, capable of delivering heterologous protective antigens, and may also provide non-specific benefits when engineered to express immunomodulatory proteins. With some improvements in the expression system, it could be used to induce a targeted immune response against specific mucosal pathogens, and co-expression of several antigens would allow development of a novel multivalent vaccine.

  16. Effects of 6/85-strain Mycoplasma gallisepticum vaccination alone at ten weeks of age or in conjunction with F-strain Mycoplasma gallisepticum inoculation overlays at twenty-two or forty-five weeks of age on the reproductive and digestive organs of commercial egg-laying hens.

    PubMed

    Viscione, K A; Branton, S L; Vance, A M; Gerard, P D; Womack, S K; Peebles, E D

    2009-03-01

    Two trials were conducted to determine the effects of a prelay 6/85-strain Mycoplasma gallisepticum (6/85MG) vaccination alone or in conjunction with time-specific F-strain M. gallisepticum (FMG) inoculation overlays on the gross reproductive and digestive organ characteristics of commercial egg-laying hens. In each trial, the following 4 treatments were applied: 1) sham vaccination at 10 wk of age; 2) vaccination of 6/85MG at 10 wk; 3) 6/85MG at 10 wk overlaid by FMG inoculation at 22 wk; and 4) 6/85MG at 10 wk overlaid by FMG at 45 wk. Two birds per isolation pen (experimental replicate unit) were necropsied at the end of both trials to observe the effects of treatment on liver weight, liver lipid and moisture concentrations, incidence of fatty liver hemorrhagic syndrome, ovary weight, mature ovarian follicle numbers, and the total and segmental weights, lengths, and histologies of the oviduct and small intestine. The applied treatments affected only liver moisture. Liver moisture content was greater in birds vaccinated with 6/85MG at 10 wk alone or in conjunction with FMG at 45 wk in comparison with sham vaccinated controls and birds that received a 6/85MG vaccination at 10 wk overlaid by an FMG inoculation at 22 wk. Prelay 6/85MG vaccinations may be a suitable substitute for prelay FMG inoculations, and FMG overlays during lay on prelay 6/85MG vaccinations may also provide continual protection against field-strain MG infections without eliciting any subsequent suppressive effects on performance, as noted in an earlier study.

  17. Competitor internal standards for quantitative detection of mycoplasma DNA.

    PubMed

    Sidhu, M K; Rashidbaigi, A; Testa, D; Liao, M J

    1995-05-01

    Homologous internal controls were used as competitor DNA in the polymerase chain reaction for the quantitative detection of mycoplasma DNA. PCR primer sets were designed on the basis of the most conserved nucleotide sequences of the 16S rRNA gene of mycoplasma species. Amplification of this gene was examined in five different mycoplasma species: Mycoplasma orale, M. hyorhinus, M. synoviae, M. gallisepticum and M. pneumoniae. To evaluate the primers, a number of different cell lines were assayed for the detection of mycoplasma infections. All positive cell lines showed a distinct product on agarose gels while uninfected cells showed no DNA amplification. Neither bacterial nor eukaryotic DNA produced any cross-reaction with the primers used, thus confirming their specificity. Internal control DNA to be used for quantitation was constructed by modifying the sizes of the wild-type amplified products and cloning them in plasmid vectors. These controls used the same primer binding sites as the wild-type and the amplified products were differentiated by a size difference. The detection limits for all the mycoplasma species by competitive quantitative PCR were estimated to range from 4 to 60 genome copies per assay as determined by ethidium bromide-stained agarose gels. These internal standards also serve as positive controls in PCR-based detection of mycoplasma DNA, and therefore accidental contamination of test samples with wild-type positive controls can be eliminated. The quantitative PCR method developed will be useful in monitoring the progression and significance of mycoplasma in the disease process.

  18. 9 CFR 113.408 - Avian mycoplasma antigen.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... ion concentration, purity, sensitivity, and specificity in accordance with the conditions prescribed... shall be tested for viable bacteria and fungi as prescribed in § 113.26. (f) Sensitivity requirements...) The sensitivity of Mycoplasma Gallisepticum Antigen shall be tested using a set of chicken and a set...

  19. Mycoplasma bovis research update

    USDA-ARS?s Scientific Manuscript database

    Mycoplasma bovis in bison is a newly emerging and potentially devastating threat to the bison industry. This bacterium is increasingly being identified, both in the United States and Canada, as the cause of severe respiratory disease outbreaks with devastating consequences for the health of the ani...

  20. Swine and Poultry Pathogens: the Complete Genome Sequences of Two Strains of Mycoplasma hyopneumoniae and a Strain of Mycoplasma synoviae†

    PubMed Central

    Vasconcelos, Ana Tereza R.; Ferreira, Henrique B.; Bizarro, Cristiano V.; Bonatto, Sandro L.; Carvalho, Marcos O.; Pinto, Paulo M.; Almeida, Darcy F.; Almeida, Luiz G. P.; Almeida, Rosana; Alves-Filho, Leonardo; Assunção, Enedina N.; Azevedo, Vasco A. C.; Bogo, Maurício R.; Brigido, Marcelo M.; Brocchi, Marcelo; Burity, Helio A.; Camargo, Anamaria A.; Camargo, Sandro S.; Carepo, Marta S.; Carraro, Dirce M.; de Mattos Cascardo, Júlio C.; Castro, Luiza A.; Cavalcanti, Gisele; Chemale, Gustavo; Collevatti, Rosane G.; Cunha, Cristina W.; Dallagiovanna, Bruno; Dambrós, Bibiana P.; Dellagostin, Odir A.; Falcão, Clarissa; Fantinatti-Garboggini, Fabiana; Felipe, Maria S. S.; Fiorentin, Laurimar; Franco, Gloria R.; Freitas, Nara S. A.; Frías, Diego; Grangeiro, Thalles B.; Grisard, Edmundo C.; Guimarães, Claudia T.; Hungria, Mariangela; Jardim, Sílvia N.; Krieger, Marco A.; Laurino, Jomar P.; Lima, Lucymara F. A.; Lopes, Maryellen I.; Loreto, Élgion L. S.; Madeira, Humberto M. F.; Manfio, Gilson P.; Maranhão, Andrea Q.; Martinkovics, Christyanne T.; Medeiros, Sílvia R. B.; Moreira, Miguel A. M.; Neiva, Márcia; Ramalho-Neto, Cicero E.; Nicolás, Marisa F.; Oliveira, Sergio C.; Paixão, Roger F. C.; Pedrosa, Fábio O.; Pena, Sérgio D. J.; Pereira, Maristela; Pereira-Ferrari, Lilian; Piffer, Itamar; Pinto, Luciano S.; Potrich, Deise P.; Salim, Anna C. M.; Santos, Fabrício R.; Schmitt, Renata; Schneider, Maria P. C.; Schrank, Augusto; Schrank, Irene S.; Schuck, Adriana F.; Seuanez, Hector N.; Silva, Denise W.; Silva, Rosane; Silva, Sérgio C.; Soares, Célia M. A.; Souza, Kelly R. L.; Souza, Rangel C.; Staats, Charley C.; Steffens, Maria B. R.; Teixeira, Santuza M. R.; Urmenyi, Turan P.; Vainstein, Marilene H.; Zuccherato, Luciana W.; Simpson, Andrew J. G.; Zaha, Arnaldo

    2005-01-01

    This work reports the results of analyses of three complete mycoplasma genomes, a pathogenic (7448) and a nonpathogenic (J) strain of the swine pathogen Mycoplasma hyopneumoniae and a strain of the avian pathogen Mycoplasma synoviae; the genome sizes of the three strains were 920,079 bp, 897,405 bp, and 799,476 bp, respectively. These genomes were compared with other sequenced mycoplasma genomes reported in the literature to examine several aspects of mycoplasma evolution. Strain-specific regions, including integrative and conjugal elements, and genome rearrangements and alterations in adhesin sequences were observed in the M. hyopneumoniae strains, and all of these were potentially related to pathogenicity. Genomic comparisons revealed that reduction in genome size implied loss of redundant metabolic pathways, with maintenance of alternative routes in different species. Horizontal gene transfer was consistently observed between M. synoviae and Mycoplasma gallisepticum. Our analyses indicated a likely transfer event of hemagglutinin-coding DNA sequences from M. gallisepticum to M. synoviae. PMID:16077101

  1. Mycoplasma infections of plants.

    PubMed

    Bove, J M

    1981-07-01

    Plants can be infected by two types of wall-less procaryotes, spiroplasmas and mycoplasma-like organisms (MLO), both located intracellularly in the phloem tissues of affected plants. Spiroplasmas have been cultured, characterized and shown to be true members of the class Mollicutes. MLO have not yet been cultured or characterized; they are thought to be mycoplasma-like on the basis of their ultrastructure as seen in situ, their sensitivity to tetracycline and resistance to penicillin. Mycoplasmas can also be found on the surface of plants. These extracellularly located organisms are members of the following genera: Spiroplasma. Mycoplasma and Acholeplasma. The presence of such surface mycoplasmas must not be overlooked when attempts to culture MLO from affected plants are undertaken. Sensitive serological techniques such as the enzyme-linked immunosorbent assay (ELISA) can successfully be used to compare the MLO located in the phloem of affected plants with those eventually cultured from the same plants. In California and Morocco periwinkles naturally infected with both Spiroplasma citri and MLO have been reported. With such doubly infected plants, the symptom expression has been that characteristic of the MLO disease (phyllody or stolbur), not that given by S. citri. Only S. citri can be cultured from such plants, but this does not indicate that S. citri is the causal agent of the disease expressed by the plant. In California many nonrutaceous plants have been found to be infected with S. citri. Stubborn affected citrus trees represent an important reservoir of S. citri, and Circulifer tenellus is an active leafhopper vector of S. citri. Hence, it is not surprising that in California MLO-infected fruit trees could also become infected with S. citri but it would not mean that S. citri is the causal agent of the disease. Criteria are discussed that are helpful in distinguishing between MLO infections and S. citri infections.

  2. Intestinal mycoplasma in Crohn's disease.

    PubMed

    Roediger, W E W

    2004-01-01

    Intestinal diversion with reconnection in active Crohn's disease (CD) indicates that luminal contents or bacteria contribute to the formation of CD lesions. Fluorescent staining for mycoplasma in freshly resected Crohn's tissue and electron microscopy reveal intracellular organisms akin to mycoplasma. Historically, tissue culture of CD has shown mycoplasma described as contaminants. Mycoplasma are surface epithelial parasites requiring exogenous cholesterol for membrane stability and cell entry. PCR of intestinal tissue has shown Mycoplasma pneumoniae to be detectable more significantly in CD. Oral M. iowae in experimental poultry localizes to the distal small bowel and colon. Hypothetically, lipopeptides of mycoplasmal membranes are proposed to cause chronicity and stronger immune responses than by other bacteria. 'Intestinal' mycoplasmas, from a number of observations, deserve consideration as organisms mediating inflammation of acute and chronic CD.

  3. Evaluation of Mycoplasma Inactivation during Production of Biologics: Egg-Based Viral Vaccines as a Model▿

    PubMed Central

    David, Selwyn A. Wilson; Volokhov, Dmitriy V.; Ye, Zhiping; Chizhikov, Vladimir

    2010-01-01

    Although mycoplasmas are generally considered to be harmless commensals, some mycoplasma species are able to cause infections in pediatric, geriatric, or immunocompromised patients. Thus, accidental contamination of biologics with mycoplasmas represents a potential risk for the health of individuals who receive cell-derived biological and pharmaceutical products. To assess the efficiency of inactivation of mycoplasmas by the agents used in the manufacture of egg-derived influenza vaccines, we carried out a series of experiments aimed at monitoring the viability of mycoplasmas spiked into both chicken allantoic fluid and protein-rich microbiological media and then treated with beta-propiolactone, formalin, cetyltrimethylammonium bromide, Triton X-100, and sodium deoxycholate, which are agents that are commonly used for virus inactivation and disruption of viral particles during influenza vaccine production. Twenty-two mycoplasma species (with one to four strains of each species) were exposed to these inactivating agents at different concentrations. The most efficient inactivation of the mycoplasmas evaluated was observed with either 0.5% Triton X-100 or 0.5% sodium deoxycholate. Cetyltrimethylammonium bromide at concentrations of ≥0.08% was also able to rapidly inactivate (in less than 30 min) all mycoplasmas tested. In contrast, negligible reductions in mycoplasma titers were observed with 0.0125 to 0.025% formaldehyde. However, increasing the concentration of formaldehyde to 0.1 to 0.2% improved the mycoplasmacidal effect. Incubation of mycoplasmas with 0.1% beta-propiolactone for 1 to 24 h had a marked mycoplasmacidal effect. A comparison of the mycoplasma inactivation profiles showed that strains of selected species (Mycoplasma synoviae, Mycoplasma gallisepticum, Mycoplasma orale, Mycoplasma pneumoniae, and Acholeplasma laidlawii) represent a set of strains that can be utilized to validate the effectiveness of mycoplasma clearance obtained by inactivation and

  4. Animal model of Mycoplasma fermentans respiratory infection

    PubMed Central

    2013-01-01

    Background Mycoplasma fermentans has been associated with respiratory, genitourinary tract infections and rheumatoid diseases but its role as pathogen is controversial. The purpose of this study was to probe that Mycoplasma fermentans is able to produce respiratory tract infection and migrate to several organs on an experimental infection model in hamsters. One hundred and twenty six hamsters were divided in six groups (A-F) of 21 hamsters each. Animals of groups A, B, C were intratracheally injected with one of the mycoplasma strains: Mycoplasma fermentans P 140 (wild strain), Mycoplasma fermentans PG 18 (type strain) or Mycoplasma pneumoniae Eaton strain. Groups D, E, F were the negative, media, and sham controls. Fragments of trachea, lungs, kidney, heart, brain and spleen were cultured and used for the histopathological study. U frequency test was used to compare recovery of mycoplasmas from organs. Results Mycoplasmas were detected by culture and PCR. The three mycoplasma strains induced an interstitial pneumonia; they also migrated to several organs and persisted there for at least 50 days. Mycoplasma fermentans P 140 induced a more severe damage in lungs than Mycoplasma fermentans PG 18. Mycoplasma pneumoniae produced severe damage in lungs and renal damage. Conclusions Mycoplasma fermentans induced a respiratory tract infection and persisted in different organs for several weeks in hamsters. This finding may help to explain the ability of Mycoplasma fermentans to induce pneumonia and chronic infectious diseases in humans. PMID:23298636

  5. [The comparison of tests for qualitative evaluation of Ureaplasma parvum, Mycoplasma hominis: "Mycoplasma duo", "Ureaplasma microtest", "Mycoplasma microtest" and "AmpliSens-Florocenosis-mycoplasma-FL"].

    PubMed

    Rumiantseva, T A; Varlamova, A V; Gushchin, A E; Bezrukov, V M

    2014-08-01

    The genital mycoplasma is an opportunistic bacteria and its detection is to be implemented in qualitative format. The study was organized to compare reagents kits "Mycoplasma Duo", "Ureaplasma Microtest", "Mycoplasma microtest" and "AmpliSens-Florocenosis-Mycoplasma-FL". The study resulted in high indicators of diagnostic sensitivity and diagnostic specificity for all kits. At that, the lowest indicators were registered under application of "Mycoplasma Duo" kit. The study reveled correlation of qualitative values detected by using cultural analysis and polymerase chain reaction. The reproducibility of qualitative values of cultural method occurred significantly lower in comparison with "AmpliSens-Florocenosis-Mycoplasma-FL " kit.

  6. Effects of supplemental dietary phytase and 25-hydroxycholecalciferol on the digestive and reproductive organ characteristics of commercial layers inoculated before or at the onset of lay with the F-strain of Mycoplasma gallisepticum.

    PubMed

    Peebles, E D; Branton, S L; Burnham, M R; Whitmarsh, S K; Gerard, P D

    2007-08-01

    In 3 trials, the effects of dietary supplementation with phytase (PHY) and 25-hydroxycholecalciferol (25-D3) on the digestive and reproductive organ characteristics of commercial layers that were inoculated prelay (12 wk of age) or at the onset of lay (22 wk of age) with F-strain Mycoplasma gallisepticum (FMG) were assessed at 58 wk of age. Experimental layer diets that included a basal control diet or a control diet supplemented with 0.025% PHY and 25-D3 were fed from 20 through 58 wk of age. As a percentage of total oviduct weight, magnum weight was lower in birds that were inoculated (sham or FMG) at lay onset compared with those that were inoculated prelay, and in FMG-inoculated birds, relative duodenum length was greater in those inoculated at 12 compared with 22 wk. Also, as percentages of organ weight or length, infundibulum length and isthmus weight were increased, whereas duodenum length was decreased by dietary supplementation with PHY and 25-D3. The overall timing (12 vs. 22 wk) of inoculation can affect the reproductive organ characteristics of layers, whereas, more specifically, the timing of an FMG inoculation may affect their digestive organ structure. Furthermore, independent of inoculation timing and type, the reproductive organ and digestive systems of laying hens may be influenced by dietary supplementation with PHY and 25-D3.

  7. Mycoplasma polysaccharide protects against complement

    PubMed Central

    Bolland, Jeffrey R.; Simmons, Warren L.; Daubenspeck, James M.

    2012-01-01

    Although they lack a cell wall, mycoplasmas do possess a glycocalyx. The interactions between the glycocalyx, mycoplasmal surface proteins and host complement were explored using the murine pathogen Mycoplasma pulmonis as a model. It was previously shown that the length of the tandem repeat region of the surface lipoprotein Vsa is associated with susceptibility to complement-mediated killing. Cells producing a long Vsa containing about 40 repeats are resistant to complement, whereas strains that produce a short Vsa of five or fewer repeats are susceptible. We show here that the length of the Vsa protein modulates the affinity of the M. pulmonis EPS-I polysaccharide for the mycoplasma cell surface, with more EPS-I being associated with mycoplasmas producing a short Vsa protein. An examination of mutants that lack EPS-I revealed that planktonic mycoplasmas were highly susceptible to complement killing even when the Vsa protein was long, demonstrating that both EPS-I and Vsa length contribute to resistance. In contrast, the mycoplasmas were resistant to complement even in the absence of EPS-I when the cells were encased in a biofilm. PMID:22504437

  8. Mycoplasma sturni from blue jays and northern mockingbirds with conjunctivitis in Florida.

    PubMed

    Ley, D H; Geary, S J; Berkhoff, J E; McLaren, J M; Levisohn, S

    1998-04-01

    Northern mockingbirds (Mimus polyglottos) and blue jays (Cyanocitta cristata) in a Florida (USA) wildlife care facility developed clinical signs and gross lesions suggestive of the ongoing outbreak of Mycoplasma gallisepticum (MG) conjunctivitis in house finches (Carpodacus mexicanus) and American goldfinches (Carduelis tristis). Mycoplasmal organisms were cultured from conjunctival/corneal swabs of birds with sinusitis, conjunctivitis, and/or epiphora. All of the isolates tested were identified as Mycoplasma sturni by indirect immunofluorescence. Mycoplasma sturni as well as MG should be considered in the differential diagnosis of songbirds with conjunctivitis.

  9. Effects of 6/85-strain Mycoplasma gallisepticum Vaccination Alone at Ten Weeks of Age or in Conjunction with F-strain M. gallisepticum Inoculation Overlays at 22 or 45 Weeks of Age on the Reproductive and Digestive....Hens.

    USDA-ARS?s Scientific Manuscript database

    Two trials were conducted to determine the effects of a prelay 6/85-strain M. gallisepticum (6/85MG) vaccination alone or in conjunction with time specific F-strain M. gallisepticum (FMG) inoculation overlays on the gross reproductive and digestive organ characteristics of commercial egg-laying hens...

  10. Effects of Time Specific F-strain Mycoplasma gallisepticum Inoculation Overlays on Prelay ts-11-strain M. gallisepticum Vaccination on Digestive and Reproductive Organ Characteristics of Commercial Egg-Laying Hens

    USDA-ARS?s Scientific Manuscript database

    Two trials were conducted to determine the effects of a prelay ts11-strain M. gallisepticum (ts11MG) vaccination alone or in conjunction with F-strain M. gallisepticum (FMG) inoculation overlays at 2 different age periods during lay on the digestive and reproductive organ characteristics of commerci...

  11. Mycoplasma tullyi sp. nov., isolated from penguins of the genus Spheniscus.

    PubMed

    Yavari, Christine A; Ramírez, Ana S; Nicholas, Robin A J; Radford, Alan D; Darby, Alistair C; Bradbury, Janet M

    2017-09-12

    A mycoplasma isolated from the liver of a dead Humboldt penguin (Spheniscus humboldti) and designated strain 56A97T, was investigated to determine its taxonomic status. Complete 16S rRNA gene sequence analysis indicated that the organism was most closely related to Mycoplasma gallisepticum and Mycoplasma imitans(99.7 and 99.9 % similarity, respectively). The average DNA-DNA hybridization values between strain 56A97T and M. gallisepticum and M. imitans were 39.5 and 30 %, respectively and the Genome to Genome Distance Calculator gave results of 29.10 and 23.50 %, respectively. The 16S-23S rRNA intergenic spacer was 72-73 % similar to M. gallisepticum strains and 52.2 % to M. imitans. A partial sequence of rpoB was 91.1-92 % similar to M. gallisepticum strains and 84.7 % to M. imitans. Colonies possessed a typical fried-egg appearance and electron micrographs revealed the lack of a cell wall and a nearly spherical morphology, with an electron-dense tip-like structure on some flask-shaped cells. The isolate required sterol for growth, fermented glucose, adsorbed and haemolysed erythrocytes, but did not hydrolyse arginine or urea. The strain was compared serologically against 110 previously described Mycoplasma reference strains, showing that, except for M. gallisepticum, strain 56A97T is not related to any of the previously described species, although weak cross-reactions were evident. Genomic information, serological reactions and phenotypic properties demonstrate that this organism represents a novel species of the genus Mycoplasma, for which the name Mycoplasma tullyi sp. nov. is proposed; the type strain is 56A97T (ATCC BAA-1432T, DSM 21909T, NCTC 11747T).

  12. Identification of Lipoprotein MslA as a Neoteric Virulence Factor of Mycoplasma gallisepticum▿

    PubMed Central

    Szczepanek, S. M.; Frasca, S.; Schumacher, V. L.; Liao, X.; Padula, M.; Djordjevic, S. P.; Geary, S. J.

    2010-01-01

    Many lipoproteins are expressed on the surfaces of mycoplasmas, and some have been implicated as playing roles in pathogenesis. Family 2 lipoproteins of Mycoplasma pneumoniae have a conserved “mycoplasma lipoprotein X” central domain and a “mycoplasma lipoprotein 10” C-terminal domain and are differentially expressed in response to environmental conditions. Homologues of family 2 lipoproteins are Mycoplasma specific and include the lipoprotein of Mycoplasma gallisepticum, encoded by the MGA0674 gene. Comparative transcriptomic analysis of the M. gallisepticum live attenuated vaccine strain F and the virulent strain Rlow, reported in this study, indicated that MGA0674 is one of several differentially expressed genes. The MGA0674-encoded lipoprotein is a proteolytically processed, immunogenic, TX-114 detergent-phase protein which appears to have antigenic divergence between field strains Rlow and S6. We examined the virulence of an Rlow ΔMGA0674 mutant (P1H9) in vivo and observed reduced recovery and attenuated virulence in the tracheas of experimentally infected chickens. The virulence of two additional Rlow ΔMGA0674 mutants, 2162 and 2204, was assessed in a second in vivo virulence experiment. These mutants exhibited partial to complete attenuation in vivo, but recovery was observed more frequently. Since only Mycoplasma species harbor homologues of MGA0674, the gene product has been renamed “Mycoplasma-specific lipoprotein A” (MslA). Collectively, these data indicate that MslA is an immunogenic lipoprotein exhibiting reduced expression in an attenuated strain and plays a role in M. gallisepticum virulence. PMID:20515935

  13. Mycoplasmas in diseases of humans.

    PubMed Central

    Embree, J E; Embil, J A

    1980-01-01

    The roles of Mycoplasma pneumoniae, M. hominis and Ureaplasma urealyticum in diseases of humans are currently under investigation. M. pneumoniae, which causes primary atypical pneumonia, is a well established pathogen of the respiratory tract. Complications of infection by this organism are also being recognized; they include disorders of the hematopoietic, cardiovascular, central nervous, musculoskeletal, cutaneous and gastrointestinal systems. The roles of the genital mycoplasmas M. hominis and U. urealyticum are controversial but may include infections of the genitourinary tract and in pregnancy as well as diseases of the newborn, such as neonatal pneumonia and meningitis. In this review atypical pneumonia due to M. pneumoniae is described and the role of mycoplasmas in other diseases is discussed. Images FIG. 1A FIG. 1B FIG. 2 PMID:6790148

  14. Comparison of multiple genes and 16S-23S rRNA intergenic space region for their capacity in high resolution melt curve analysis to differentiate Mycoplasma gallisepticum vaccine strain ts-11 from field strains.

    PubMed

    Ghorashi, Seyed A; Bradbury, Janet M; Ferguson-Noel, Naola M; Noormohammadi, Amir H

    2013-12-27

    Mycoplasma gallisepticum (MG) is an important avian pathogen causing significant economic losses in the global poultry industry. In an attempt to compare and evaluate existing genotyping methods for differentiation of MG strains/isolates, high resolution melt (HRM) curve analysis was applied to 5 different PCR methods targeting vlhA, pvpA, gapA, mgc2 genes and 16S-23S rRNA intergenic space region (IGSR). To assess the discriminatory power of PCR-HRM of examined genes and IGSR, MG strains ts-11, F, 6/85 and S6, and, initially, 8 field isolates were tested. All MG strains/isolates were differentiated using PCR-HRM curve analysis and genotype confidence percentage (GCP) values of vlhA and pvpA genes, while only 0, 3 and 4 out of 12 MG strains/isolates were differentiated using gapA, mgc2 genes and IGSR, respectively. The HRM curve analysis of vlhA and pvpA genes was found to be highly correlated with the genetic diversity of the targeted genes confirmed by sequence analysis of amplicons generated from MG strains. The potential of the vlhA and pvpA genes was also demonstrated for genotyping of 12 additional MG strains from Europe and the USA. Results from this study provide a direct comparison between genes previously used in sequencing-based genotyping methods for MG strain identification and highlight the usefulness of vlhA and pvpA HRM curve analyses as rapid and reliable tools specially for diagnosis and differentiation of MG strains used here.

  15. Pathobiology of Mycoplasma suis.

    PubMed

    Hoelzle, Ludwig E; Zeder, Michael; Felder, Kathrin M; Hoelzle, Katharina

    2014-10-01

    Mycoplasma suis is an uncultivable bacterium lacking a cell wall that attaches to and may invade the red blood cells of pigs. M. suis infections occur worldwide and cause the pig industry serious economic losses due to the disease known as infectious anaemia of pigs or, historically, porcine eperythrozoonosis. Infectious anaemia of pigs is characterised predominantly by acute haemolytic or chronic anaemia, along with non-specific manifestations, such as growth retardation in feeder pigs and poor reproductive performance in sows. The fastidious nature of M. suis, as well as the lack of an in vitro cultivation system, has hampered the understanding of the biology and pathogenicity of this organism. Pathogenetic mechanisms of M. suis include direct destruction of red blood cells by adhesion, invasion, nutrient scavenging, immune-mediated lysis and eryptosis, as well as endothelial targeting. Recently published genome sequences, in combination with proteome analyses, have generated new insights into the pathogenicity of M. suis. The present review combines these data with the knowledge provided by experimental M. suis infections.

  16. Motility of Mycoplasma pneumoniae.

    PubMed Central

    Radestock, U; Bredt, W

    1977-01-01

    Cell of Mycoplasma pneumoniae FH gliding on a glass surface in liquid medium were examined by microscopic observation and quantitatively by microcinematography (30 frames per min). Comparisons were made only within the individual experiments. The cells moved in an irregular pattern with numerous narrow bends and circles. They never changed their leading end. The average speed (without pauses) was relatively constant between o.2 and 0.5 mum/s. The maximum speed was about 1.5 to 2.0 mum/s. The movements were interrupted by resting periods of different lengths and frequency. Temperature, viscosity, pH, and the presence of yeast extract in the medium influenced the motility significantly; changes in glucose, calcium ions, and serum content were less effective. The movements were affected by iodoacetate, p-mercuribenzoate, and mitomycin C at inhibitory or subinhibitory concentrations. Sodium fluoride, sodium cyanide, dinitrophenol, chloramphenicol, puromycin, cholchicin, and cytochalasin B at minimal inhibitory concentrations did not affect motility. The movements were effectively inhibited by anti-M. pneumoniae antiserum. Studies with absorbed antiserum suggested that the surface components involved in motility are heat labile. The gliding of M. pneumoniae cells required an intact energy metabolism and the proteins involved seemed to have a low turnover. Images PMID:14925

  17. Mycoplasma infections in small ruminants.

    PubMed

    Ruffin, D C

    2001-07-01

    Mycoplasmas have complex mechanisms of antigenic variation that allow them to evade the immune system. These organisms cause a variety of clinical syndromes that can have a significant economic effect on small ruminant production. The syndromes range from acute septicemia and death to chronic infection resulting in decreased production. Recent research findings have shed light on the means by which these organisms evade the host immune response and cause or contribute to the development of disease in the host. This article provides a review of the pathogenesis, clinical signs, and treatment options for common disease syndromes involving Mycoplasma spp. in small ruminants.

  18. Impact of fowlpox-vectored Mycoplasma gallisepticum vaccine Vectormune® FP MG on layer hen egg production and egg quality parameters

    USDA-ARS?s Scientific Manuscript database

    This study was conducted to determine the impact of vaccination with Vectormune®FP MG on egg production and egg quality characteristics of white leghorn hens. Due to questions of the efficacy of this vaccine in preventing M. gallisepticum mediated pathology, the ability of this vaccine to protect a...

  19. Non-occurrence of Mycoplasma genitalium in clinical specimens.

    PubMed

    Samra, Z; Borin, M; Bukowsky, Y; Lipshitz, Y; Sompolinsky, D

    1988-02-01

    Five hundred and thirteen clinical specimens, mainly from patients with urogenital inflammations, were examined for Ureaplasma urealyticum and mycoplasmas, including cultures for Mycoplasma genitalium. The study yielded 95 isolates of Ureaplasma urealyticum, 37 isolates of Mycoplasma hominis and two isolates of Mycoplasma fermentans, but no growth of Mycoplasma genitalium was obtained. It was concluded that Mycoplasma genitalium is a relatively rare inhabitant of the human urogenital tract in Israel.

  20. Draft Genome Sequence of "Candidatus Mycoplasma haemobos," a Hemotropic Mycoplasma Identified in Cattle in Mexico.

    PubMed

    Martínez-Ocampo, Fernando; Rodríguez-Camarillo, Sergio D; Amaro-Estrada, Itzel; Quiroz-Castañeda, Rosa Estela

    2016-07-07

    We present here the draft genome sequence of the first "Candidatus Mycoplasma haemobos" strain found in cattle in Mexico. This hemotropic mycoplasma causes acute and chronic disease in animals. This genome is a starting point for studying the role of this mycoplasma in coinfections and synergistic mechanisms associated with the disease.

  1. Survey of campylobacter, salmonella and mycoplasmas in house crows (Corvus splendens) in Malaysia.

    PubMed

    Ganapathy, K; Saleha, A A; Jaganathan, M; Tan, C G; Chong, C T; Tang, S C; Ideris, A; Dare, C M; Bradbury, J M

    2007-05-05

    House crows (Corvus splendens) in Selangor, Malaysia were examined for the presence of Campylobacter species, Salmonella species, Mycoplasma gallisepticum and Mycoplasma synoviae by serology, culture and pcr. For the detection of Campylobacter and Salmonella species swabs were taken either from the intestine or cloaca. For the detection of mycoplasmas, swabs were taken either from the choanal cleft or trachea for culture and pcr and serum samples were tested by the rapid serum agglutination (rsa) and monoclonal antibody-blocking elisa (mbelisa) for antibodies to M gallisepticum and M synoviae. For campylobacter, 25.3 per cent of the crows were positive by culture, and the species identified were Campylobacter jejuni and Campylobacter coli. No Salmonella species were isolated. Four of 24 swabs were positive for M gallisepticum dna but none gave positive results for M synoviae dna. No M gallisepticum or M synoviae antibodies were detected by rsa but 60 per cent of the sera gave positive reactions for M gallisepticum and 13 per cent gave positive reactions for M synoviae by mbelisa.

  2. Reduction of hydrogen peroxide accumulation and toxicity by a catalase from Mycoplasma iowae.

    PubMed

    Pritchard, Rachel E; Prassinos, Alexandre J; Osborne, John D; Raviv, Ziv; Balish, Mitchell F

    2014-01-01

    Mycoplasma iowae is a well-established avian pathogen that can infect and damage many sites throughout the body. One potential mediator of cellular damage by mycoplasmas is the production of H2O2 via a glycerol catabolic pathway whose genes are widespread amongst many mycoplasma species. Previous sequencing of M. iowae serovar I strain 695 revealed the presence of not only genes for H2O2 production through glycerol catabolism but also the first documented mycoplasma gene for catalase, which degrades H2O2. To test the activity of M. iowae catalase in degrading H2O2, we studied catalase activity and H2O2 accumulation by both M. iowae serovar K strain DK-CPA, whose genome we sequenced, and strains of the H2O2-producing species Mycoplasma gallisepticum engineered to produce M. iowae catalase by transformation with the M. iowae putative catalase gene, katE. H2O2-mediated virulence by M. iowae serovar K and catalase-producing M. gallisepticum transformants were also analyzed using a Caenorhabditis elegans toxicity assay, which has never previously been used in conjunction with mycoplasmas. We found that M. iowae katE encodes an active catalase that, when expressed in M. gallisepticum, reduces both the amount of H2O2 produced and the amount of damage to C. elegans in the presence of glycerol. Therefore, the correlation between the presence of glycerol catabolism genes and the use of H2O2 as a virulence factor by mycoplasmas might not be absolute.

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

    PubMed

    Bébéar, C M; de Barbeyrac, B; Pereyre, S; Renaudin, H; Clerc, M; Bébéar, C

    2008-08-01

    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 mycoplasmas. Moxifloxacin killed the 30 mycoplasma isolates tested at a concentration

  4. The characterization of Mycoplasma synoviae EF-Tu protein and proteins involved in hemadherence and their N-terminal amino acid sequences.

    PubMed

    Bencina, D; Narat, M; Dovc, P; Drobnic-Valic, M; Habe, F; Kleven, S H

    1999-04-01

    An abundant cytoplasmic 43-kDa protein from Mycoplasma synoviae, a major pathogen from poultry, was identified as elongation factor Tu. The N-terminal amino acid sequence (AKLDFDRSKEHVNVGTIGHV) has 90% identity with the sequence of the Mycoplasma hominis elongation factor Tu protein. Monoclonal antibodies reacting with the M. synoviae elongation factor Tu protein also reacted with 43-kDa proteins from the avian Mycoplasma species Mycoplasma gallinarum, Mycoplasma gallinaceum, Mycoplasma pullorum, Mycoplasma cloacale, Mycoplasma iners and Mycoplasma meleagridis, but not with the proteins from Mycoplasma gallisepticum, Mycoplasma imitans or Mycoplasma iowae. In addition, two groups of phase variable integral membrane proteins, pMSA and pMSB, associated with hemadherence and pathogenicity of M. synoviae strains AAY-4 and ULB925 were identified. The cleavage of a larger hemagglutinating protein encoded by a gene homologous to the vlhA gene of M. synoviae generates pMSB1 and pMSA1 proteins defined by mAb 125 and by hemagglutination inhibiting mAb 3E10, respectively. The N-terminal amino acid sequences of pMSA proteins (SENKLI ... and SENETQ ...) probably indicate the cleavage site of the M. synoviae strain ULB 925 hemagglutinin.

  5. Prevention and Detection of Mycoplasma Contamination in Cell Culture

    PubMed Central

    Nikfarjam, Laleh; Farzaneh, Parvaneh

    2012-01-01

    One of the main problems in cell culture is mycoplasma infection. It can extensively affect cell physiology and metabolism. As the applications of cell culture increase in research, industrial production and cell therapy, more concerns about mycoplasma contamination and detection will arise. This review will provide valuable information about: 1. the ways in which cells are contaminated and the frequency and source of mycoplasma species in cell culture; 2. the ways to prevent mycoplasma contamination in cell culture; 3. the importance of mycoplasma tests in cell culture; 4. different methods to identify mycoplasma contamination; 5. the consequences of mycoplasma contamination in cell culture and 6. available methods to eliminate mycoplasma contamination. Awareness about the sources of mycoplasma and pursuing aseptic techniques in cell culture along with reliable detection methods of mycoplasma contamination can provide an appropriate situation to prevent mycoplasma contamination in cell culture. PMID:23508237

  6. Mycoplasma infection of ducks and geese.

    PubMed

    Stipkovits, L; Szathmary, S

    2012-11-01

    Production of ducks and geese in certain parts of the world is very important. Mycoplasma diseases cause significant losses to the duck and goose industry. This review summarizes the epidemiological, clinical, and pathomorphological characteristics of mycoplasma diseases of ducks and geese and the involvement of the various mycoplasma species in their pathogenesis. The role of mycoplasma infections in the development of clinical signs, pathological lesions, and mortality of challenged birds is demonstrated in challenge experiments. Transmission of mycoplasma in the ovary and eggs resulting in the reduction of egg production and an increase of embryo mortality has been shown in challenge experiments as well as in field studies. The susceptibility of many mycoplasma isolates of the most important mycoplasma species of duck and goose origin were tested and showed relatively high average minimum inhibitory concentrations of lincomycin, tilosin, oxytetracycline, chlortetracycline, and enrofloxacin but not for tiamulin. The successful treatment of mycoplasma infections with antibiotics in ducks and geese should be selected based on the minimum inhibitory concentration values against the mycoplasmas isolated from the flock.

  7. House Finch (Haemorhous mexicanus) Conjunctivitis, and Mycoplasma spp. Isolated from North American Wild Birds, 1994–2015

    PubMed Central

    Ley, David H.; Hawley, Dana M.; Geary, Steven J.; Dhondt, André A.

    2016-01-01

    Sampling wild birds for mycoplasma culture has been key to the study of ‘House Finch (Haemorhous mexicanus) conjunctivitis’, yielding isolates of Mycoplasma gallisepticum spanning the temporal and geographic ranges of disease from emergence to endemicity. Faced with the challenges and costs of sample collection over time and from remote locations for submission to our laboratory for mycoplasma culture, protocols evolved to achieve a practical optimum. Herein we report making M. gallisepticum isolates from House Finches almost every year since the disease emerged in 1994, and we now have 227 isolates from 17 US states. Our wild bird host range for M. gallisepticum isolates includes Blue Jay (Cyanocitta cristata), American Goldfinch (Spinus tristis), Lesser Goldfinch (Spinus psaltria), Purple Finch (Haemorhous purpureus), Evening Grosbeak (Coccothraustes vespertinus), and herein first reports for Western Scrub-jay (Aphelocoma californica), and American Crow (Corvus brachyrhynchos). By collecting and identifying isolates from birds with clinical signs similar to ‘house finch conjunctivitis’ we also expanded the known host range of Mycoplasma sturni and obtained isolates from additional wild bird species. Accumulating evidence shows that a diverse range of wild bird species may carry or have been exposed to M. gallisepticum in the US, as in Europe and Asia. Therefore, the emergence of a pathogenic M. gallisepticum strain in House Finches may actually be the exception that has allowed us to identify the broader epidemiologic picture. PMID:27285414

  8. House Finch (Haemorhous mexicanus) Conjunctivitis, and Mycoplasma spp. Isolated from North American Wild Birds, 1994-2015.

    PubMed

    Ley, David H; Hawley, Dana M; Geary, Steven J; Dhondt, André A

    2016-07-01

    Sampling wild birds for mycoplasma culture has been key to the study of House Finch (Haemorhous mexicanus) conjunctivitis, yielding isolates of Mycoplasma gallisepticum spanning the temporal and geographic ranges of disease from emergence to endemicity. Faced with the challenges and costs of sample collection over time and from remote locations for submission to our laboratory for mycoplasma culture, protocols evolved to achieve a practical optimum. Herein we report making M. gallisepticum isolates from House Finches almost every year since the disease emerged in 1994, and we now have 227 isolates from 17 states. Our wild bird host range for M. gallisepticum isolates includes Blue Jay ( Cyanocitta cristata ), American Goldfinch (Spinus tristis), Lesser Goldfinch (Spinus psaltria), Purple Finch (Haemorhous purpureus), Evening Grosbeak ( Coccothraustes vespertinus ), and herein first reports for Western Scrub-jay ( Aphelocoma californica ), and American Crow ( Corvus brachyrhynchos ). By collecting and identifying isolates from birds with clinical signs similar to those of House Finch conjunctivitis, we also expanded the known host range of Mycoplasma sturni and obtained isolates from additional wild bird species. Accumulating evidence shows that a diverse range of wild bird species may carry or have been exposed to M. gallisepticum in the US, as in Europe and Asia. Therefore, the emergence of a pathogenic M. gallisepticum strain in House Finches may actually be the exception that has allowed us to identify the broader epidemiologic picture.

  9. Mycoplasma genitalium mg200 and mg386 genes are involved in gliding motility but not in cytadherence.

    PubMed

    Pich, Oscar Q; Burgos, Raul; Ferrer-Navarro, Mario; Querol, Enrique; Piñol, Jaume

    2006-06-01

    Isolation and characterization of transposon-generated Mycoplasma genitalium gliding-deficient mutants has implicated mg200 and mg386 genes in gliding motility. The proposed role of these genes was confirmed by restoration of the gliding phenotype in deficient mutants through gene complementation with their respective mg386 or mg200 wild-type copies. mg200 and mg386 are the first reported gliding-associated mycoplasma genes not directly involved in cytadherence. Orthologues of MG200 and MG386 proteins are also found in the slow gliding mycoplasmas, Mycoplasma pneumoniae and Mycoplasma gallisepticum, suggesting the existence of a unique set of proteins involved in slow gliding motility. MG200 and MG386 proteins share common features, such as the presence of enriched in aromatic and glycine residues boxes and an acidic and proline-rich domain, suggesting that these motifs could play a significant role in gliding motility.

  10. Molecular Biology and Pathogenicity of Mycoplasmas

    PubMed Central

    Razin, Shmuel; Yogev, David; Naot, Yehudith

    1998-01-01

    The recent sequencing of the entire genomes of Mycoplasma genitalium and M. pneumoniae has attracted considerable attention to the molecular biology of mycoplasmas, the smallest self-replicating organisms. It appears that we are now much closer to the goal of defining, in molecular terms, the entire machinery of a self-replicating cell. Comparative genomics based on comparison of the genomic makeup of mycoplasmal genomes with those of other bacteria, has opened new ways of looking at the evolutionary history of the mycoplasmas. There is now solid genetic support for the hypothesis that mycoplasmas have evolved as a branch of gram-positive bacteria by a process of reductive evolution. During this process, the mycoplasmas lost considerable portions of their ancestors’ chromosomes but retained the genes essential for life. Thus, the mycoplasmal genomes carry a high percentage of conserved genes, greatly facilitating gene annotation. The significant genome compaction that occurred in mycoplasmas was made possible by adopting a parasitic mode of life. The supply of nutrients from their hosts apparently enabled mycoplasmas to lose, during evolution, the genes for many assimilative processes. During their evolution and adaptation to a parasitic mode of life, the mycoplasmas have developed various genetic systems providing a highly plastic set of variable surface proteins to evade the host immune system. The uniqueness of the mycoplasmal systems is manifested by the presence of highly mutable modules combined with an ability to expand the antigenic repertoire by generating structural alternatives, all compressed into limited genomic sequences. In the absence of a cell wall and a periplasmic space, the majority of surface variable antigens in mycoplasmas are lipoproteins. Apart from providing specific antimycoplasmal defense, the host immune system is also involved in the development of pathogenic lesions and exacerbation of mycoplasma induced diseases. Mycoplasmas are

  11. Mycoplasmas, plants, insect vectors: a matrimonial triangle.

    PubMed

    Garnier, M; Foissac, X; Gaurivaud, P; Laigret, F; Renaudin, J; Saillard, C; Bové, J M

    2001-10-01

    Plant pathogenic mycoplasmas were discovered by electron microscopy, in 1967, long after the discovery and culture in 1898 of the first pathogenic mycoplasma of animal origin, Mycoplasma mycoides. Mycoplasmas are Eubacteria of the class Mollicutes, a group of organisms phylogenetically related to Gram-positive bacteria. Their more characteristic features reside in the small size of their genomes, the low guanine (G) plus cytosine (C) content of their genomic DNA and the lack of a cell wall. Plant pathogenic mycoplasmas are responsible for several hundred diseases and belong to two groups: the phytoplasmas and the spiroplasmas. The phytoplasmas (previously called MLOs, for mycoplasma like organisms) were discovered first; they are pleiomorphic, and have so far resisted in vitro cultivation. Phytoplasmas represent the largest group of plant pathogenic Mollicutes. Only three plant pathogenic spiroplasmas are known today. Spiroplasma citri, the agent of citrus stubborn was discovered and cultured in 1970 and shown to be helical and motile. S. kunkelii is the causal agent of corn stunt. S. phoeniceum, responsible for periwinkle yellows, was discovered in Syria. There are many other spiroplasmas associated with insects and ticks. Plant pathogenic mycoplasmas are restricted to the phloem sieve tubes in which circulates the photosynthetically-enriched sap, the food for many phloem-feeding insects (aphids, leafhoppers, psyllids, etc.). Interestingly, phytopathogenic mycoplasmas are very specifically transmitted by leafhoppers or psyllid species. In this paper, the most recent knowledge on phytopathogenic mycoplasmas in relation with their insect and plant habitats is presented as well as the experiments carried out to control plant mycoplasma diseases, by expression of mycoplasma-directed-antibodies in plants (plantibodies).

  12. Characteristics of Mycoplasma hominis adhesion.

    PubMed Central

    Olson, L D; Gilbert, A A

    1993-01-01

    Mycoplasma hominis, a human pathogen, has previously been observed to bind to sulfatide separated on thin-layer chromatograms. It has not been demonstrated, however, that the binding is not simply a nonspecific ionic interaction. The ability of a low-passage patient isolate of M. hominis to adhere to glycoconjugates other than sulfatide and the characteristics of its binding to sulfatide were studied. Mycoplasmas were found to bind strongly and specifically in a temperature- and dose-dependent manner to only sulfatide of all of the glycolipids and glycoproteins tested. The avidity and specificity of binding, as well as the ability to inhibit the interaction specifically, suggest that the receptors to which M. hominis binds, particularly in the human urogenital tract, from which it is frequently isolated, are primarily, if not solely, sulfated glycolipids. Images PMID:8491739

  13. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 7 2014-04-01 2014-04-01 false Test for Mycoplasma. 610.30 Section 610.30 Food... GENERAL BIOLOGICAL PRODUCTS STANDARDS Mycoplasma § 610.30 Test for Mycoplasma. Except as provided... tested for the presence of Mycoplasma, as follows: Samples of the virus for this test shall be...

  14. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Test for Mycoplasma. 610.30 Section 610.30 Food... GENERAL BIOLOGICAL PRODUCTS STANDARDS Mycoplasma § 610.30 Test for Mycoplasma. Except as provided... tested for the presence of Mycoplasma, as follows: Samples of the virus for this test shall be...

  15. Mycoplasma genitalium in Toronto, Ont

    PubMed Central

    Gesink, Dionne; Racey, C. Sarai; Seah, Christine; Zittermann, Sandra; Mitterni, Leo; Juzkiw, Jerry; Jamieson, Heather; Greer, Jane; Singh, Sudesh; Jensen, Jørgen Skov; Allen, Vanessa

    2016-01-01

    Objective To estimate the prevalence of Mycoplasma genitalium in Toronto, Ont; detect mutations associated with macrolide and fluoroquinolone resistance; and describe treatment outcomes. Design Prospective, cross-sectional study. Setting A sexual health clinic in Toronto. Participants A consecutive sample of men and women attending the sexual health clinic between September 1, 2013, and December 20, 2013. Interventions Participants underwent testing for M genitalium, along with standard sexually transmitted infection screening. All samples that had positive results for M genitalium were tested for mutations associated with resistance to macrolides and fluoroquinolones. Mycoplasma genitalium treatment was based on resistance profile and verified with a test of cure. Main outcome measures Positive results for M genitalium and antibiotic resistance. Results A total of 1193 men and women participated in the study. Overall, 4.5% of the 884 men and 3.2% of the 309 women had positive test results for M genitalium. Asymptomatic infection was common (52.0%). Macrolide resistance–mediating mutations were found in 58.0% of the M genitalium infections. No treatment failure was observed for azithromycin-treated cases. Treatment failure was suspected for 16.7% of cases treated with moxifloxacin. Conclusion Mycoplasma genitalium is present in Canada, with a prevalence comparable to chlamydia and gonorrhea, and has high macrolide and fluoroquinolone resistance. PMID:27331225

  16. Submasseteric abscess caused by Mycoplasma salivarium infection.

    PubMed

    Grisold, Andrea J; Hoenigl, Martin; Leitner, Eva; Jakse, Klaus; Feierl, Gebhard; Raggam, Reinhard B; Marth, Egon

    2008-11-01

    Mycoplasma salivarium preferentially resides in the human oral cavity. Unlike other Mycoplasma species, M. salivarium has not been regarded as a pathogen, although one case of M. salivarium-caused arthritis in a patient with hypogammaglobulinemia has been reported. We describe the first case of submasseteric abscess caused by M. salivarium.

  17. Bilateral optic papillitis following mycoplasma pneumoniae pneumonia.

    PubMed

    Milla, E; Zografos, L; Piguet, B

    1998-01-01

    Mycoplasma pneumoniae is an atypical bacterium that can cause a great variety of respiratory infections and be responsible for ocular involvement such as conjunctivitis, anterior uveitis and very rarely optic neuropathy. We report herein an additional case of bilateral optic disc swelling with profound visual loss following Mycoplasma pneumoniae pneumonia and review the world literature on the ocular manifestations associated with this pathogen.

  18. One test microbial diagnostic microarray for identification of Mycoplasma mycoides subsp. mycoides and other Mycoplasma species.

    PubMed

    Tonelli, A; Sacchini, F; Krasteva, I; Zilli, K; Scacchia, M; Beaurepaire, C; Nantel, A; Pini, A

    2012-11-01

    The present study describes the use of microarray technology for rapid identification and differentiation of Mycoplasma mycoides subsp. mycoides from other mycoplasmas that may be pathogenic to ruminants, including those of the Mycoplasma mycoides cluster, genetically and antigenically strictly correlated with Mycoplasma mycoides subsp. mycoides. A microarray containing genetic sequences of 55 different bacterial species from Acholeplasma, Mycoplasma, Spiroplasma and Ureaplasma genera was constructed. Sequences to genes of interest were collected in FASTA format from NCBI. The collected sequences were processed with OligoPicker software. Oligonucleotides were then checked for their selectivity with BLAST searches in GenBank. The microarray was tested with ATCC/NCTC strains of Mycoplasma spp. of veterinary importance in ruminants including Mycoplasma belonging to the mycoides cluster as well as Mycoplasma mycoides subsp. mycoides and Mycoplasma mycoides subsp. capri field strains. The results showed that but one ATCC/NCTC reference strains hybridized with their species-specific sequences showed a profile/signature different and distinct from each other. The heat-map of the hybridization results for the nine genes interrogated for Mycoplasma mycoides subsp. mycoides demonstrated that the reference strain Mycoplasma mycoides subsp mycoides PG1 was positive for all of the gene sequences spotted on the microarray. CBPP field, vaccine and reference strains were all typed to be M. mycoides subsp. mycoides, and seven of the nine strains gave positive hybridization results for all of the nine genes. Two Italian strains were negative for some of the genes. Comparison with non-Mycoplasma mycoides subsp. mycoides reference strains showed some positive signals or considerable homology to Mycoplasma mycoides subsp. mycoides genes. As expected, some correlations were observed between the strictly genetically and antigenically correlated Mycoplasma mycoides subsp. mycoides and

  19. Eradication of Mycoplasma contaminations from cell cultures.

    PubMed

    Uphoff, Cord C; Drexler, Hans G

    2014-04-14

    Mycoplasma contaminations have a multitude of effects on cultured cell lines that may influence the results of experiments or pollute bioactive substances isolated from the eukaryotic cells. The elimination of mycoplasma contaminations from cell cultures with antibiotics has been proven to be a practical alternative to discarding and re-establishing important or irreplaceable cell lines. Different fluoroquinolones, tetracyclins, pleuromutilins, and macrolides shown to have strong anti-mycoplasma properties are employed for the decontamination. These antibiotics are applied as single treatments, as combination treatment of two antibiotics in parallel or successively, or in combination with a surface-active peptide to enhance the action of the antibiotic. The protocols in this unit allow eradication of mycoplasmas, prevention of the development of resistant mycoplasma strains, and potential cure of heavily contaminated and damaged cells. Consistent and permanent alterations to eukaryotic cells attributable to the treatment have not been demonstrated.

  20. Mycoplasma penetrans and Other Mycoplasmas in Urine of Human Immunodeficiency Virus-Positive Children

    PubMed Central

    Hussain, Althaf I.; Robson, William Lane M.; Kelley, Robin; Reid, Tanya; Gangemi, J. David

    1999-01-01

    Urine samples from children with human immunodeficiency virus (HIV) infection and healthy controls were examined for mycoplasmas by culture. Standard biochemical assays, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and PCR (16S and 16S-23S spacer rRNA region) were used for identification of isolates. Mycoplasmas were identified from 13 (87%) of 15 HIV-positive patients and 3 (20%) of 15 HIV-negative control patients. The frequency and type of mycoplasma varied with the severity of HIV infection. Mycoplasma penetrans, Mycoplasma pirum, Mycoplasma fermentans, and Mycoplasma genitalium were isolated from patients with severe immunodeficiency. Mycoplasma hominis and Ureaplasma urealyticum were isolated more frequently from children in the early stages of HIV infection and from HIV-negative patients. Mycoplasma penetrans was isolated from one (50%) of two patients in Centers for Disease Control and Prevention (CDC) group B and from five (55.5%) of nine pediatric patients with AIDS (CDC group C). This is the first report that indicates that “AIDS-associated” mycoplasmas are more common in HIV-infected children than in HIV-negative controls. PMID:10203515

  1. Co-infection with Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' in three cats from Brazil.

    PubMed

    de Morais, Helio A; Guimarães, Ana Marcia S; Vidotto, Odilon; Baumann, Aline; Biondo, Alexander W; Messick, Joanne B

    2007-12-01

    The two most common haemotropic Mycoplasma of cats, Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' have been identified using molecular techniques in all continents, except Antarctica. We report the first molecular characterization in South America of a dual infection with M haemofelis and 'Candidatus Mycoplasma haemominutum' in three domestic cats. The 16S ribosomal RNA gene was amplified in three anaemic cats in which haemoplasma organisms were seen attached to the erythrocytes in the peripheral blood smear. Bands of the expected size for M haemofelis and 'Candidatus Mycoplasma haemominutum' were observed in all three cats. The 393 bp segment of one of the amplicons had a similarity value of 100% to M haemofelis, whereas the other amplicon, a 192 bp segment, was 100% similar to 'Candidatus Mycoplasma haemominutum'. After diagnosis, two cats received blood transfusion and they were all treated with doxycycline. All three cats recovered uneventfully.

  2. Detection of Mycoplasma canadense and Mycoplasma californicum in dairy cattle from Argentina.

    PubMed

    Tamiozzo, Pablo J; Estanguet, Abel A; Maito, Julia; Tirante, Liliana; Pol, Martin; Giraudo, José A

    2014-01-01

    Different species of Mycoplasma can affect bovine cattle, causing several diseases. PCR sequencing and further analysis of the 16S-23S rRNA ITS region have shown a significant interspecies variability among Mollicutes. Sixteen suspected isolates of Mycoplasma spp. obtained from milk samples from dairy herds were amplified (16S-23S rRNA ITS region). Fourteen out of those 16 suspected Mycoplasma spp. isolates were PCR-positive. To confirm the identity of Mycoplasma bovis, these 14 isolates were tested by another species-specific PCR. Seven of the isolates rendered a positive result. The products of 16S-23S rRNA ITS PCR from one isolate that was identified as M. bovis and from two other isolates, identified as non- M. bovis were randomly selected, sequenced and analyzed. The three sequences (A, B and C) showed 100% similarity with M. bovis, Mycoplasma canadense and Mycoplasma californicum respectively.

  3. Sedimentation counting and morphology of Mycoplasma.

    PubMed

    Clark, H W

    1965-11-01

    Clark, Harold W. (The George Washington University School of Medicine, Washington, D.C.). Sedimentation counting and morphology of Mycoplasma. J. Bacteriol. 90:1373-1386. 1965.-The sedimentation technique for counting viral particles was applied to the quantitation and morphological identification of Mycoplasma in broth cultures. Mycoplasma, apparently in their native form, firmly adhered to the surface, when sedimented on glass cover slips or onto electron microscope grids. The sedimented cover slip preparations stained with crystal violet could be readily counted in the light microscope. The cultures sedimented onto electron microscope grids were readily counted at low magnification and provided excellent preparations for morphological examination at higher magnifications. It was found that air-dried Mycoplasma particles were enlarged considerably because of excessive flattening. Fixation of sedimented Mycoplasma particles in diluted OsO(4) prior to air drying yielded a more realistic morphology, with various sizes and shapes in the stages of the growth cycle exhibited. A new technique of differentially staining Mycoplasma colonies on agar plates was developed to facilitate the quantitation of viable colony-forming units for comparison with total counts. The use of plastic or Parafilm gaskets for dry mounting was developed to facilitate the handling and examination of the stained cover slip preparations. The results of this investigation indicated that the growth cycle of some Mycoplasma species includes a stage of hexadic fission with the cleavage of minimal reproductive units (less than 100 mmu) containing a limited deoxyribonucleic acid genetic coding molecule (approximately 4 x 10(6)).

  4. Gliding Direction of Mycoplasma mobile

    PubMed Central

    Morio, Hanako; Kasai, Taishi

    2015-01-01

    ABSTRACT Mycoplasma mobile glides in the direction of its cell pole by a unique mechanism in which hundreds of legs, each protruding from its own gliding unit, catch, pull, and release sialylated oligosaccharides fixed on a solid surface. In this study, we found that 77% of cells glided to the left with a change in direction of 8.4° ± 17.6° μm−1 displacement. The cell body did not roll around the cell axis, and elongated, thinner cells also glided while tracing a curved trajectory to the left. Under viscous conditions, the range of deviation of the gliding direction decreased. In the presence of 250 μM free sialyllactose, in which the binding of the legs (i.e., the catching of sialylated oligosaccharides) was reduced, 70% and 30% of cells glided to the left and the right, respectively, with changes in direction of ∼30° μm−1. The gliding ghosts, in which a cell was permeabilized by Triton X-100 and reactivated by ATP, glided more straightly. These results can be explained by the following assumptions based on the suggested gliding machinery and mechanism: (i) the units of gliding machinery may be aligned helically around the cell, (ii) the legs extend via the process of thermal fluctuation and catch the sialylated oligosaccharides, and (iii) the legs generate a propulsion force that is tilted from the cell axis to the left in 70% and to the right in 30% of cells. IMPORTANCE Mycoplasmas are bacteria that are generally parasitic to animals and plants. Some Mycoplasma species form a protrusion at a pole, bind to solid surfaces, and glide. Although these species appear to consistently glide in the direction of the protrusion, their exact gliding direction has not been examined. This study analyzed the gliding direction in detail under various conditions and, based on the results, suggested features of the machinery and the mechanism of gliding. PMID:26503848

  5. 9 CFR 147.31 - Laboratory procedures recommended for the real-time polymerase chain reaction test for Mycoplasma...

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... test for Mycoplasma gallisepticum (MGLP ReTi). (a) DNA extraction. Use Qiagen Qiamp Mini Kit for DNA extraction or equivalent validated technique/procedure. This kit utilizes the following methods: 100 µl of swab suspension incubates with 10 µl of proteinase K and 400 µl of lysis buffer at 56 °C for 10...

  6. 9 CFR 147.31 - Laboratory procedures recommended for the real-time polymerase chain reaction test for Mycoplasma...

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... test for Mycoplasma gallisepticum (MGLP ReTi). (a) DNA extraction. Use Qiagen Qiamp Mini Kit for DNA extraction or equivalent validated technique/procedure. This kit utilizes the following methods: 100 µl of swab suspension incubates with 10 µl of proteinase K and 400 µl of lysis buffer at 56 °C for 10...

  7. 9 CFR 147.31 - Laboratory procedures recommended for the real-time polymerase chain reaction test for Mycoplasma...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... the real-time polymerase chain reaction test for Mycoplasma gallisepticum (MGLP ReTi). 147.31 Section... Examination Procedures § 147.31 Laboratory procedures recommended for the real-time polymerase chain reaction... value) was determined to be the PCR cycle number at which the fluorescence of the reaction exceeded 30...

  8. 9 CFR 147.30 - Laboratory procedure recommended for the polymerase chain reaction (PCR) test for Mycoplasma...

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... the polymerase chain reaction (PCR) test for Mycoplasma gallisepticum and M. synoviae. 147.30 Section... Examination Procedures § 147.30 Laboratory procedure recommended for the polymerase chain reaction (PCR) test... should consist of the following sequences: ER12JA07.005 (c) Polymerase chain reaction. (1) Treat...

  9. 9 CFR 147.30 - Laboratory procedure recommended for the polymerase chain reaction (PCR) test for Mycoplasma...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... the polymerase chain reaction (PCR) test for Mycoplasma gallisepticum and M. synoviae. 147.30 Section... Examination Procedures § 147.30 Laboratory procedure recommended for the polymerase chain reaction (PCR) test... should consist of the following sequences: ER12JA07.005 (c) Polymerase chain reaction. (1) Treat...

  10. 9 CFR 147.30 - Laboratory procedure recommended for the polymerase chain reaction (PCR) test for Mycoplasma...

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... the polymerase chain reaction (PCR) test for Mycoplasma gallisepticum and M. synoviae. 147.30 Section... Examination Procedures § 147.30 Laboratory procedure recommended for the polymerase chain reaction (PCR) test... should consist of the following sequences: ER12JA07.005 (c) Polymerase chain reaction. (1) Treat...

  11. 9 CFR 147.30 - Laboratory procedure recommended for the polymerase chain reaction (PCR) test for Mycoplasma...

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... the polymerase chain reaction (PCR) test for Mycoplasma gallisepticum and M. synoviae. 147.30 Section... Examination Procedures § 147.30 Laboratory procedure recommended for the polymerase chain reaction (PCR) test... should consist of the following sequences: ER12JA07.005 (c) Polymerase chain reaction. (1) Treat...

  12. 9 CFR 147.30 - Laboratory procedure recommended for the polymerase chain reaction (PCR) test for Mycoplasma...

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... the polymerase chain reaction (PCR) test for Mycoplasma gallisepticum and M. synoviae. 147.30 Section... Examination Procedures § 147.30 Laboratory procedure recommended for the polymerase chain reaction (PCR) test... should consist of the following sequences: ER12JA07.005 (c) Polymerase chain reaction. (1) Treat...

  13. 9 CFR 147.31 - Laboratory procedures recommended for the real-time polymerase chain reaction test for Mycoplasma...

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... the real-time polymerase chain reaction test for Mycoplasma gallisepticum (MGLP ReTi). 147.31 Section... Examination Procedures § 147.31 Laboratory procedures recommended for the real-time polymerase chain reaction... lp gene. (c) MGLP ReTi. Primers and probe should be utilized in a 25 µl reaction containing 12.5...

  14. 9 CFR 147.31 - Laboratory procedures recommended for the real-time polymerase chain reaction test for Mycoplasma...

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... the real-time polymerase chain reaction test for Mycoplasma gallisepticum (MGLP ReTi). 147.31 Section... Examination Procedures § 147.31 Laboratory procedures recommended for the real-time polymerase chain reaction... lp gene. (c) MGLP ReTi. Primers and probe should be utilized in a 25 µl reaction containing 12.5...

  15. Optimized PCR-based detection of mycoplasma.

    PubMed

    Dobrovolny, Paige L; Bess, Dan

    2011-06-20

    The maintenance of contamination-free cell lines is essential to cell-based research. Among the biggest contaminant concerns are mycoplasma contamination. Although mycoplasma do not usually kill contaminated cells, they are difficult to detect and can cause a variety of effects on cultured cells, including altered metabolism, slowed proliferation and chromosomal aberrations. In short, mycoplasma contamination compromises the value of those cell lines in providing accurate data for life science research. The sources of mycoplasma contamination in the laboratory are very challenging to completely control. As certain mycoplasma species are found on human skin, they can be introduced through poor aseptic technique. Additionally, they can come from contaminated supplements such as fetal bovine serum, and most importantly from other contaminated cell cultures. Once mycoplasma contaminates a culture, it can quickly spread to contaminate other areas of the lab. Strict adherence to good laboratory practices such as good aseptic technique are key, and routine testing for mycoplasma is highly recommended for successful control of mycoplasma contamination. PCR-based detection of mycoplasma has become a very popular method for routine cell line maintenance. PCR-based detection methods are highly sensitive and can provide rapid results, which allows researchers to respond quickly to isolate and eliminate contamination once it is detected in comparison to the time required using microbiological techniques. The LookOut Mycoplasma PCR Detection Kit is highly sensitive, with a detection limit of only 2 genomes per μl. Taking advantage of the highly specific JumpStart Taq DNA Polymerase and a proprietary primer design, false positives are greatly reduced. The convenient 8-tube format, strips pre-coated with dNTPs, and associated primers helps increase the throughput to meet the needs of customers with larger collections of cell lines. Given the extreme sensitivity of the kit, great

  16. A College Epidemic of Mycoplasma Pneumoniae.

    ERIC Educational Resources Information Center

    Ralston, David; Cochran, Burt

    1979-01-01

    The article reports on an outbreak of mycoplasma pneumoniae at the California Polytechnic State University including a historical background of the disease, clinical features, laboratory findings for treated patients, treatment, and clinical clues for diagnosis. (JMF)

  17. Mycoplasma penetrans bacteremia and primary antiphospholipid syndrome.

    PubMed Central

    Yáñez, A.; Cedillo, L.; Neyrolles, O.; Alonso, E.; Prévost, M. C.; Rojas, J.; Watson, H. L.; Blanchard, A.; Cassell, G. H.

    1999-01-01

    Mycoplasma penetrans, a rare bacterium so far only found in HIV-infected persons, was isolated in the blood and throat of a non-HIV-infected patient with primary antiphospholipid syndrome (whose etiology and pathogenesis are unknown). PMID:10081687

  18. Survey of plasmids in various mycoplasmas.

    PubMed Central

    Harasawa, R.; Barile, M. F.

    1983-01-01

    Thirty-three strains representing 15 distinct Mycoplasma, Acholeplasma, and Spiroplasma species were examined for the presence of plasmid DNA by agarose gel electrophoresis. The electrophoretic patterns of the DNAs of three strains, Mycoplasma sp. strain 747, Spiroplasma mirum strain SMCA, and M. hominis strain 1257, suggested the presence of a plasmid with molecular weights of approximately 70, 10, and 9 megadaltons, respectively. The functions of these plasmids are currently unknown. Images FIG. 1 PMID:6679154

  19. Detection and prevention of mycoplasma hominis infection

    DOEpatents

    DelVecchio, Vito G.; Gallia, Gary L.; McCleskey, Ferne K.

    1997-01-21

    The present invention is directed to a rapid and sensitive method for detecting Mycoplasma hominis using M. hominis-specific probes, oligonucleotides or antibodies. In particular a target sequence can be amplified by in vitro nucleic acid amplification techniques, detected by nucleic acid hybridization using the subject probes and oligonucleotides or detected by immunoassay using M. hominis-specific antibodies. M. hominis-specific nucleic acids which do not recognize or hybridize to genomic nucleic acid of other Mycoplasma species are also provided.

  20. Mycoplasmas and Ureaplasmas as Neonatal Pathogens

    PubMed Central

    Waites, Ken B.; Katz, Brenda; Schelonka, Robert L.

    2005-01-01

    The genital mycoplasmas represent a complex and unique group of microorganisms that have been associated with a wide array of infectious diseases in adults and infants. The lack of conclusive knowledge regarding the pathogenic potential of Mycoplasma and Ureaplasma spp. in many conditions is due to a general unfamiliarity of physicians and microbiology laboratories with their fastidious growth requirements, leading to difficulty in their detection; their high prevalence in healthy persons; the poor design of research studies attempting to base association with disease on the mere presence of the organisms in the lower urogenital tract; the failure to consider multifactorial aspects of diseases; and considering these genital mycoplasmas only as a last resort. The situation is now changing because of a greater appreciation of the genital mycoplasmas as perinatal pathogens and improvements in laboratory detection, particularly with regard to the development of powerful molecular nucleic acid amplification tests. This review summarizes the epidemiology of genital mycoplasmas as causes of neonatal infections and premature birth; evidence linking ureaplasmas with bronchopulmonary dysplasia; recent changes in the taxonomy of the genus Ureaplasma; the neonatal host response to mycoplasma and ureaplasma infections; advances in laboratory detection, including molecular methods; and therapeutic considerations for treatment of systemic diseases. PMID:16223956

  1. Molecular detection of Mycoplasma wenyonii and 'Candidatus Mycoplasma haemobos' in cattle in Hokkaido, Japan.

    PubMed

    Tagawa, Michihito; Matsumoto, Kotaro; Inokuma, Hisashi

    2008-11-25

    Blood samples from 78 cattle were tested for hemoplasma infection using molecular methods. PCR and sequence analysis revealed that 17 cattle were infected with Mycoplasma wenyonii, while 13 were infected with 'Candidatus Mycoplasma haemobos'. Four animals were infected with both species. This is the first study to report hemoplasma species infection among cattle in Japan.

  2. Genome Sequence of Mycoplasma hyorhinis Isolated from Cell Cultures

    PubMed Central

    Cibulski, Samuel Paulo; Siqueira, Franciele Maboni; Teixeira, Thais Fumaco; Mayer, Fabiana Quoos; Almeida, Luiz Gonzaga

    2016-01-01

    Mycoplasmas are major contaminants of mammalian cell cultures. Here, the complete genome sequence of Mycoplasma hyorhinis recovered from Madin-Darby bovine kidney (MDBK) cells is reported. PMID:27738034

  3. [Mycoplasma Pneumoniae-Induced Meningoencephalitis].

    PubMed

    Özel, C; Dafotakis, M; Nikoubashman, O; Litmathe, J; Matz, O; Schöne, U

    2015-07-01

    In clinical practice, secondary infections of the central nervous system (CNS) represent rare yet severe complications of their respective primary infections. In this case report, we describe a 22-year-old patient with a medical history of Asthma bronchiale, who developed significant neurological deficits after a respiratory infection. The neurological symptoms progressed despite antibiotic therapy with vancomycin, ampicillin and ceftriaxone. The patient's cerebrospinal fluid and a cranial magnetic resonance imaging (MRI) furnished evidence of acute meningoencephalitis. Microbiological assessment confirmed an acute mycoplasma pneumonia infection. Changing the patient's antibiotic regimen to minocycline and prednisolone led to significant clinical improvement. Pathomechanisms and therapeutic options to treat meningoencephalitis will be discussed in the following. © Georg Thieme Verlag KG Stuttgart · New York.

  4. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Test for Mycoplasma. 610.30 Section 610.30 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS GENERAL BIOLOGICAL PRODUCTS STANDARDS Mycoplasma § 610.30 Test for Mycoplasma. Except as provided...

  5. Draft Genome Sequence of “Candidatus Mycoplasma haemobos,” a Hemotropic Mycoplasma Identified in Cattle in Mexico

    PubMed Central

    Martínez-Ocampo, Fernando; Rodríguez-Camarillo, Sergio D.; Amaro-Estrada, Itzel

    2016-01-01

    We present here the draft genome sequence of the first “Candidatus Mycoplasma haemobos” strain found in cattle in Mexico. This hemotropic mycoplasma causes acute and chronic disease in animals. This genome is a starting point for studying the role of this mycoplasma in coinfections and synergistic mechanisms associated with the disease. PMID:27389272

  6. Genes Found Essential in Other Mycoplasmas Are Dispensable in Mycoplasma bovis

    PubMed Central

    Sharma, Shukriti; Markham, Philip F.; Browning, Glenn F.

    2014-01-01

    Mycoplasmas are regarded to be useful models for studying the minimum genetic complement required for independent survival of an organism. Mycoplasma bovis is a globally distributed pathogen causing pneumonia, mastitis, arthritis, otitis media and reproductive tract disease, and genome sequences of three strains, the type strain PG45 and two strains isolated in China, have been published. In this study, several Tn4001 based transposon constructs were generated and used to create a M. bovis PG45 insertional mutant library. Direct genome sequencing of 319 independent insertions detected disruptions in 129 genes in M. bovis, 48 of which had homologues in Mycoplasma mycoides subspecies mycoides SC and 99 of which had homologues in Mycoplasma agalactiae. Sixteen genes found to be essential in previous studies on other mycoplasma species were found to be dispensable. Five of these genes have previously been predicted to be part of the core set of 153 essential genes in mycoplasmas. Thus this study has extended the list of non-essential genes of mycoplasmas from that previously generated by studies in other species. PMID:24897538

  7. Eosinophilic Fasciitis Associated with Mycoplasma arginini Infection

    PubMed Central

    Silló, Pálma; Pintér, Dóra; Ostorházi, Eszter; Mazán, Mercedes; Wikonkál, Norbert; Pónyai, Katinka; Volokhov, Dmitriy V.; Chizhikov, Vladimir E.; Szathmary, Susan; Stipkovits, Laszlo

    2012-01-01

    Eosinophilic fasciitis (EF) with generalized sclerodermiform skin lesions developed over a 19-month period in a previously healthy 23-year-old man. Although we confirmed EF by skin histology and laboratory tests, the recurrent fevers and the clinical observation of sclerotic prepuce with urethritis indicated further bacteriological analysis by conventional microbiological and DNA-based tests. Urethra cultures were positive for an arginine-hydrolyzing mycoplasma and Ureaplasma urealyticum. The patient also had serum IgM antibodies to Mycoplasma pneumoniae using enzyme-linked immunosorbent assay (ELISA)-based qualitative detection. Mycoplasma arginini was isolated from two independent venous blood serum samples and was identified by conventional microbiological tests and sequencing of the 16S rRNA and rpoB genes (GenBank sequence accession numbers HM179555 and HM179556, respectively). M. arginini genomic DNA also was detected by species-specific PCR in the skin lesion biopsy sample. Treatment with corticosteroids and long-term courses of selected antibiotics led to remission of skin symptoms and normalization of laboratory values. This report provides the first evidence of EF associated with mycoplasma infection and the second report of human infection with M. arginini and therefore suggests that this mycoplasma infection might have contributed to the pathogenesis of the disease. PMID:22189109

  8. Mycoplasma mastitis: causes, transmission, and control.

    PubMed

    Fox, Lawrence K

    2012-07-01

    Mycoplasma mastitis is an emerging mastitis pathogen. Herd prevalence has increased over the past decade, and this increase parallels the increase in average dairy herd size. It has been documented that the importation of cattle into a herd can result in new cases of Mycoplasma disease in general and Mycoplasma mastitis specifically. Thus, expanding herds are likely to have a greater incidence of this disease. Transmission of the agent can result from either contact with diseased animals or with colonized or asymptomatically infected cattle. Initial transmission might occur via nose-to-nose contact and result in an outbreak of Mycoplasma mastitis, or it might occur during the milking time. This would suggest that new, incoming animals should be quarantined before being comingled with original herd animals. Quarantining does not seem to be a biosecurity strategy often practiced in control of Mycoplasma mastitis and may not be warranted in herds with excellent milking time hygiene practices. The ability to monitor for the incipient stages of an outbreak, often done through bulk tank milk culturing, is recommended.

  9. Identification of cross-reactive antigens between Mycoplasma pulmonis and Mycoplasma arthritidis.

    PubMed Central

    Minion, F C; Brown, M B; Cassell, G H

    1984-01-01

    Serological cross-reactivity between Mycoplasma pulmonis and Mycoplasma arthritidis was investigated by enzyme-linked immunosorbent assay, immunoanalysis of electrophoretic blots, and protein A immunoprecipitation reactions. The results demonstrate that one-way cross-reactivity was present in both hyperimmunized and naturally infected rats and that the predominant cross-reactive antigens were M. pulmonis surface proteins. Distinct immunoblot patterns were demonstrated for M. pulmonis and M. arthritidis, allowing differentiation of the two species. The response to M. arthritidis antigens during natural infections differed greatly from that during hyperimmunization. Evidence suggested that nonprotein antigens were major determinants eliciting the antibody response to this mycoplasma. Images PMID:6690399

  10. Occurrence of mycoplasmas in semen samples of birds of prey.

    PubMed

    Lierz, M; Hafez, H M

    2008-10-01

    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.

  11. The occurrence of mycoplasmas in selected wild North American waterfowl

    USGS Publications Warehouse

    Goldberg, D.R.; Samuel, M.D.; Thomas, C.B.; Sharp, P.; Krapu, G.L.; Robb, J.R.

    1995-01-01

    We determined the prevalence of mycoplasma infection in breeding mallard (Anas platyrhynchos) and canvasback (Aythya valisineria) hens and their broods from the central United States (1988 to 1990); and wintering American black duck (Anas rubripes) and mallard hens from the eastern United States (1990 to 1993). Mycoplasmas were isolated by culturing tracheal swabs from 656 live birds and tissue samples from 112 dead waterfowl. Nine (18%) of 51 mycoplasma isolates were identified as Mycoplasma anatis; M. anatis was recovered from four mallards, a black duck, and a gadwall (Anas strepera) duckling. Nineteen (37%) of 51 mycoplasma isolates were identified as Mycoplasma cloacale; these isolates were obtained from mallard, canvasback, and black duck adults, and from a mallard duckling. Additional unspeciated mycoplasmas were isolated from mallards, black ducks, and one canvasback.

  12. Dialysis Culture of T-Strain Mycoplasmas

    PubMed Central

    Masover, Gerald K.; Hayflick, Leonard

    1974-01-01

    Using dialyzing cultures of T-strain mycoplasmas, it was possible to make some observations relevant to the growth and metabolism of these organisms which would not be possible in nondialyzing cultures due to growth inhibition of the organisms by elevated pH and increased ammonium ion concentration in media containing urea. The rate of ammonia accumulation was found to be related to the initial urea concentration in the medium and could not be accounted for by any change in the multiplication rate of the organisms. More ammonia was generated than could be accounted for by the added urea alone, suggesting that an ammonia-producing activity other than urease may be present in T-strain mycoplasmas. Titers above 107 color change units per ml were achieved in dialysis cultures of a T-strain mycoplasma in the presence of urea, and such titers were maintained for approximately 60 h during dialysis culture in the absence of added urea. PMID:4595203

  13. EXPERIMENTAL INFECTION WITH MYCOPLASMA PNEUMONIAE (EATON'S AGENT)

    PubMed Central

    Dajani, Adnan S.; Clyde, Wallace A.; Denny, Floyd W.

    1965-01-01

    The pathogenesis of Mycoplasma pneumoniae infection was studied in the Syrian hamster with qualitative and quantitative culture methods and special histopathologic techniques. The animals were readily infected with the mycoplasma, which multiplied throughout the respiratory tract. Sensitivity of this experimental host to infection was indicated by the 50 per cent infective dose, which was 10 colony-forming units of the organism. Inoculation consistently resulted in the production of peribronchial pneumonitis which was induced by the mycoplasma. The organisms were visualized in a superficial location in the mucosa of involved bronchi, by means of indirect fluorescent antibody staining and by a modification of the Brown and Brenn technique. The data indicate applicability of the hamster to the study of problems concerned with M. pneumoniae disease which are impractical or impossible to resolve in the human host. PMID:14319403

  14. Insights into the pathogenesis of Mycoplasma pneumoniae

    PubMed Central

    He, Jun; Liu, Mihua; Ye, Zhufeng; Tan, Tianping; Liu, Xinghui; You, Xiaoxing; Zeng, Yanhua; Wu, Yimou

    2016-01-01

    Mycoplasma are the smallest prokaryotic microbes present in nature. These wall-less, malleable organisms can pass through cell filters, and grow and propagate under cell-free conditions in vitro. Of the pathogenic Mycoplasma Mycoplasma pneumoniae has been examined the most. In addition to primary atypical pneumonia and community-acquired pneumonia with predominantly respiratory symptoms, M. pneumoniae can also induce autoimmune hemolytic anemia and other diseases in the blood, cardiovascular system, gastrointestinal tract and skin, and can induce pericarditis, myocarditis, nephritis and meningitis. The pathogenesis of M. pneumoniae infection is complex and remains to be fully elucidated. The present review aimed to summarize several direct damage mechanisms, including adhesion damage, destruction of membrane fusion, nutrition depletion, invasive damage, toxic damage, inflammatory damage and immune damage. Further investigations are required for determining the detailed pathogenesis of M. pneumoniae. PMID:27667580

  15. Sialidase Activity in Mycoplasma synoviae

    PubMed Central

    May, Meghan; Kleven, Stanley H.; Brown, Daniel R.

    2008-01-01

    SUMMARY Eleven strains of the avian pathogen Mycoplasma synoviae were evaluated for the presence of sialidase activity by using the fluorogenic substrate 2′-(4-methylumbelliferyl)-α-D-N-acetylneuraminic acid and the sialidase inhibitor 2-deoxy-2,3-didehydro-N-acetylneuraminic acid. The kinetics of in vitro growth in modified Frey’s medium were also assessed for each strain. Five strains had been isolated from clinically symptomatic chickens, and strains WVU1853T and K3344 have been demonstrated to be capable of reproducing disease in specific pathogen-free chickens. All strains exhibited sialidase activity, although the amount varied 65 fold (P < 0.0001) from 1.3 x 10−7 to 2.0 x 10−9 activity units/colony-forming unit among strains. Strains originally isolated from clinically symptomatic birds had more (P < 0.05) sialidase activity than strains from asymptomatic birds. Strain WVU1853T exhibited the most sialidase activity (P < 0.0001) and grew to the highest culture density (P < 0.0001) among strains, but across strains the rank correlation of growth rate with sialidase activity was not significant. Negligible activity was detected in conditioned culture supernatant fluid. This is the first report of sialidase activity in pathogenic strains of M. synoviae, which suggests a potential enzymatic basis for virulence of the organism. PMID:18251389

  16. Mycoplasma feriruminatoris sp. nov., a fast growing Mycoplasma species isolated from wild Caprinae.

    PubMed

    Jores, Joerg; Fischer, Anne; Sirand-Pugnet, Pascal; Thomann, Andreas; Liebler-Tenorio, Elisabeth M; Schnee, Christiane; Santana-Cruz, Ivette; Heller, Martin; Frey, Joachim

    2013-12-01

    Five Mycoplasma strains from wild Caprinae were analyzed: four from Alpine ibex (Capra ibex) which died at the Berlin Zoo between 1993 and 1994, one from a Rocky Mountain goat collected in the USA prior to 1987. These five strains represented a population different from the populations belonging to the 'Mycoplasma mycoides cluster' as tested using multi locus sequence typing, Matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis and DNA-DNA hybridization. Analysis of the 16S rRNA gene (rrs), genomic sequence based in silico as well as laboratory DNA-DNA hybridization, and the analysis of phenotypic traits in particular their exceptionally rapid growth all confirmed that they do not belong to any Mycoplasma species described to date. We therefore suggest these strains represent a novel species, for which we propose the name Mycoplasma feriruminatoris sp. nov. The type strain is G5847(T) (=DSM 26019(T)=NCTC 13622(T)) [corrected].

  17. Candidatus Mycoplasma haematoparvum and Mycoplasma haemocanis infections in dogs from the United States.

    PubMed

    Compton, S M; Maggi, R G; Breitschwerdt, E B

    2012-12-01

    Mycoplasma haemocanis (Mhc) and Candidatus Mycoplasma haematoparvum (CMhp) have been described in dogs. Historically, microscopic visualization of hemotropic Mycoplasma spp. has occurred most often in immunocompromised or splenectomized dogs. The aim of this study was to determine the Mhc and CMhp prevalences among dogs from the United States. Novel 16S rRNA and RNAseP gene PCR assays were used to amplify hemotropic Mycoplasma species DNA for GenBank sequence alignment. Among the study population, hemoplasma prevalence was 1.3% (7 out of 506), with Mhc and CMhp prevalences of 0.6% and 0.8%, respectively. Two of six CMhp-infected dogs were co-infected with a Bartonella sp., and a third dog was seroreactive to Bartonella henselae antigens. The prevalence of Mhc and CMhp in this study was low; potential blood donors should be screened; and dogs and people can be co-infected with hemoplasma and Bartonella spp.

  18. Demonstration of neuraminidase activity in Mycoplasma neurolyticum and of neuraminidase proteins in three canine Mycoplasma species.

    PubMed

    Berčič, Rebeka Lucijana; Cizelj, Ivanka; Benčina, Mateja; Narat, Mojca; Bradbury, Janet M; Dovč, Peter; Benčina, Dušan

    2012-03-23

    Neuraminidases are virulence factors in many pathogenic microorganisms. They are present also in some Mycoplasma species that cause disease in birds, dogs and alligators. Thirty-seven Mycoplasma species have been examined previously for neuraminidase (sialidase) activity, whereas many of the species causing disease in man, ruminants, pigs, rodents and other animals have not. In this study neuraminidase enzymatic activity (NEAC) was examined in 45 previously untested Mycoplasma species, including those causing diseases in man, farm animals and laboratory animals. The only species in which NEAC was found was Mycoplasma neurolyticum, specifically, its type strain (Type A(T)) which is capable of inducing neurologic signs in inoculated young mice and rats. The NEAC of washed cells was relatively weak, but it differed even more than 10-fold among cells of cultures derived from individual colonies of M. neurolyticum. A weak NEAC was also detected in the supernatant of the M. neurolyticum broth culture. Canine Mycoplasma spp. with high sialidase activity reported previously, Mycoplasma canis, Mycoplasma cynos and Mycoplasma molare had 100-fold more NEAC than M. neurolyticum, but apparent differences in NEAC levels existed among strains of M. canis and of M. cynos. Zymograms using neuraminidase-specific chromogenic substrate were used to show proteins having NEAC. In M. canis (a field isolate Larissa and the type strain PG14(T)), M. cynos (isolate 896) and M. molare (type strain H542(T)) proteins with NEAC had molecular masses of ∼130kDa, 105kDa and 110kDa, respectively. Identification of these neuraminidases could provide the basis for their molecular characterization.

  19. Conditions for growing Mycoplasma canadense and Mycoplasma verecundum in a serum-free medium.

    PubMed

    Muñoz, G; Sotomayor, P

    1990-07-01

    Mycoplasma canadense and Mycoplasma verecundum were cultured in a serum-free medium containing bovine serum albumin, cholesterol, oleic acid, and palmitic acid in order to avoid the addition of horse serum. Growth was detected by measurement of A640 and by colony formation. The level of growth attained in this medium was less than that obtained in the horse serum-supplemented media, but colonies retained their distinctive morphology.

  20. Conditions for growing Mycoplasma canadense and Mycoplasma verecundum in a serum-free medium.

    PubMed Central

    Muñoz, G; Sotomayor, P

    1990-01-01

    Mycoplasma canadense and Mycoplasma verecundum were cultured in a serum-free medium containing bovine serum albumin, cholesterol, oleic acid, and palmitic acid in order to avoid the addition of horse serum. Growth was detected by measurement of A640 and by colony formation. The level of growth attained in this medium was less than that obtained in the horse serum-supplemented media, but colonies retained their distinctive morphology. Images PMID:2202260

  1. Identification of two pathogenic avian mycoplasmas as strains of Mycoplasma pullorum.

    PubMed

    Moalic, P Y; Kempf, I; Gesbert, F; Laigret, F

    1997-01-01

    Two mycoplasma strains were isolated from dead turkey embryos. Growth properties, biochemical and serological characteristics, and protein profiles indicated that these strains were closely related to Mycoplasma pullorum CKKT (T = type strain). This was confirmed by 16S rRNA sequence analysis, and the phylogenetic position of M. pullorum was established. Pathogenicity studies showed that the two strains, as well as M. pullorum CKKT, induced a statistically significant level of mortality after inoculation into chicken embryos.

  2. A phylogenetic analysis of the mycoplasmas: basis for their classification.

    PubMed Central

    Weisburg, W G; Tully, J G; Rose, D L; Petzel, J P; Oyaizu, H; Yang, D; Mandelco, L; Sechrest, J; Lawrence, T G; Van Etten, J

    1989-01-01

    Small-subunit rRNA sequences were determined for almost 50 species of mycoplasmas and their walled relatives, providing the basis for a phylogenetic systematic analysis of these organisms. Five groups of mycoplasmas per se were recognized (provisional names are given): the hominis group (which included species such as Mycoplasma hominis, Mycoplasma lipophilum, Mycoplasma pulmonis, and Mycoplasma neurolyticum), the pneumoniae group (which included species such as Mycoplasma pneumoniae and Mycoplasma muris), the spiroplasma group (which included species such as Mycoplasma mycoides, Spiroplasma citri, and Spiroplasma apis), the anaeroplasma group (which encompassed the anaeroplasmas and acholeplasmas), and a group known to contain only the isolated species Asteroleplasma anaerobium. In addition to these five mycoplasma groups, a sixth group of variously named gram-positive, walled organisms (which included lactobacilli, clostridia, and other organisms) was also included in the overall phylogenetic unit. In each of these six primary groups, subgroups were readily recognized and defined. Although the phylogenetic units identified by rRNA comparisons are difficult to recognize on the basis of mutually exclusive phenotypic characters alone, phenotypic justification can be given a posteriori for a number of them. PMID:2592342

  3. Identification of mycoplasma membrane proteins by systematic Tn phoA mutagenesis of a recombinant library.

    PubMed

    Cleavinger, C M; Kim, M F; Im, J H; Wise, K S

    1995-10-01

    Wall-less prokaryotes in the genus Mycoplasma include over 90 species of infectious agents whose pathogenicity for humans and other animals is currently being assessed. Molecular characterization of surface proteins is critical in this regard but is hampered by the lack of genetic systems in these organisms. We used TnphoA transposition to systematically mutagenize, in Escherichia coli, a genomic plasmid library constructed from Mycoplasma fermentans, a potential human pathogen. The strategy circumvented problems of expressing mycoplasma genes containing UGA (Trp) codons and relied on the construction of the vector pG7ZCW, designed to reduce TnphoA transposition into vector sequences. Functional phoA gene fusions directly identified genes encoding 19 putative membrane-associated proteins of M. fermentans. Sequences of fusion constructs defined three types of export sequence: (1) non-cleavable, membrane-spanning sequences, (2) signal peptides with signal peptidase (SPase) I-like cleavage sites, and (3) signal peptides with SPase II-like lipoprotein-cleavage sites which, like most other mycoplasmal lipoprotein signals analysed to date, differed from those in several Gram-negative and Gram-positive eubacteria in their lack of a Leu residue at the -3 position. Antibodies to synthetic peptides that were deduced from two fusions to predicted lipoproteins, identified corresponding amphiphilic membrane proteins of 57 kDa and 78 kDa expressed in the mycoplasma. The P57 sequence contained a proline-rich N-terminal region analogous to an adhesin of Mycoplasma gallisepticum. The P78 protein was identical to a serologically defined phase-variant surface lipoprotein. TnphoA mutagenesis provides an efficient means of systematically characterizing functionally diverse lipoproteins and other exported proteins in mycoplasmas.

  4. Chronic "Candidatus Mycoplasma turicensis" infection.

    PubMed

    Novacco, Marilisa; Boretti, Felicitas S; Wolf-Jäckel, Godelind A; Riond, Barbara; Meli, Marina L; Willi, Barbara; Lutz, Hans; Hofmann-Lehmann, Regina

    2011-04-20

    "Candidatus Mycoplasma turicensis" infects felids. The pathogenesis of "Candidatus M. turicensis" chronic infection is poorly understood. The goals of the present study were to (1) induce reactivation of the infection in chronic carrier cats by attempted immunosuppression, (2) identify potential tissue sequestration using real-time TaqMan® PCR and (3) monitor the humoral immune response by DnaK enzyme-linked immunosorbent assay (ELISA). Ten specified pathogen-free cats that had ostensibly recovered from experimental "Candidatus M. turicensis" infection were used: five cats (group 1) received high dose methylprednisolone (attempted immunosuppression), while five cats served as untreated controls (group 2). Besides weekly blood samples, tissue samples were collected from bone marrow, kidney, liver and salivary glands at selected time points. The cats in group 1 had significantly lower lymphocyte counts and higher blood glucose levels after methylprednisolone administration than the controls. After methylprednisolone administration one blood and three tissue samples from cats in group 1 tested PCR-positive; before the administration, only one sample was positive. All other samples tested PCR-negative. All cats stayed seropositive; the antibody levels of the cats in group 1 showed a significant transient decrease after methylprednisolone administration. This is the first study to report the presence of "Candidatus M. turicensis" in tissues of chronically infected cats and the persistence of anti-feline hemoplasma antibodies in the absence of detectable bacteremia. Methylprednisolone administration did not lead to a significant reactivation of the infection. Our results enhance the knowledge of "Candidatus M. turicensis" infection pathogenesis and are clinically relevant to the prognosis of hemoplasma-infected cats.

  5. Cellular Microbiology of Mycoplasma canis

    PubMed Central

    Michaels, Dina L.; Leibowitz, Jeffrey A.; Azaiza, Mohammed T.; Shil, Pollob K.; Shama, Suzanne M.; Kutish, Gerald F.; Distelhorst, Steven L.; Balish, Mitchell F.; May, Meghan A.

    2016-01-01

    Mycoplasma canis can infect many mammalian hosts but is best known as a commensal or opportunistic pathogen of dogs. The unexpected presence of M. canis in brains of dogs with idiopathic meningoencephalitis prompted new in vitro studies to help fill the void of basic knowledge about the organism's candidate virulence factors, the host responses that it elicits, and its potential roles in pathogenesis. Secretion of reactive oxygen species and sialidase varied quantitatively (P < 0.01) among strains of M. canis isolated from canine brain tissue or mucosal surfaces. All strains colonized the surface of canine MDCK epithelial and DH82 histiocyte cells and murine C8-D1A astrocytes. Transit through MDCK and DH82 cells was demonstrated by gentamicin protection assays and three-dimensional immunofluorescence imaging. Strains further varied (P < 0.01) in the extents to which they influenced the secretion of tumor necrosis factor alpha (TNF-α) and the neuroendocrine regulatory peptide endothelin-1 by DH82 cells. Inoculation with M. canis also decreased major histocompatibility complex class II (MHC-II) antigen expression by DH82 cells (P < 0.01), while secretion of gamma interferon (IFN-γ), interleukin-6 (IL-6), interleukin-10 (IL-10), and complement factor H was unaffected. The basis for differences in the responses elicited by these strains was not obvious in their genome sequences. No acute cytopathic effects on any homogeneous cell line, or consistent patterns of M. canis polyvalent antigen distribution in canine meningoencephalitis case brain tissues, were apparent. Thus, while it is not likely a primary neuropathogen, M. canis has the capacity to influence meningoencephalitis through complex interactions within the multicellular and neurochemical in vivo milieu. PMID:27045036

  6. A Compendium for Mycoplasma pneumoniae

    PubMed Central

    Parrott, Gretchen L.; Kinjo, Takeshi; Fujita, Jiro

    2016-01-01

    Historically, atypical pneumonia was a term used to describe an unusual presentation of pneumonia. Currently, it is used to describe the multitude of symptoms juxtaposing the classic symptoms found in cases of pneumococcal pneumonia. Specifically, atypical pneumonia is a syndrome resulting from a relatively common group of pathogens including Chlamydophila sp., and Mycoplasma pneumoniae. The incidence of M. pneumoniae pneumonia in adults is less than the burden experienced by children. Transmission rates among families indicate children may act as a reservoir and maintain contagiousness over a long period of time ranging from months to years. In adults, M. pneumoniae typically produces a mild, “walking” pneumonia and is considered to be one of the causes of persistent cough in patients. M. pneumoniae has also been shown to trigger the exacerbation of other lung diseases. It has been repeatedly detected in patients with bronchitis, asthma, chronic obstructive pulmonary disorder, and cystic fibrosis. Recent advances in technology allow for the rapid diagnosis of M. pneumoniae through the use of polymerase chain reaction or rapid antigen tests. With this, more effort has been afforded to identify the causative etiologic agent in all cases of pneumonia. However, previous practices, including the overprescribing of macrolide treatment in China and Japan, have created increased incidence of macrolide-resistant M. pneumoniae. Reports from these countries indicate that >85% of M. pneumoniae pneumonia pediatric cases are macrolide-resistant. Despite its extensively studied past, the smallest bacterial species still inspires some of the largest questions. The developments in microbiology, diagnostic features and techniques, epidemiology, treatment and vaccines, and upper respiratory conditions associated with M. pneumoniae in adult populations are included within this review. PMID:27148202

  7. A Compendium for Mycoplasma pneumoniae.

    PubMed

    Parrott, Gretchen L; Kinjo, Takeshi; Fujita, Jiro

    2016-01-01

    Historically, atypical pneumonia was a term used to describe an unusual presentation of pneumonia. Currently, it is used to describe the multitude of symptoms juxtaposing the classic symptoms found in cases of pneumococcal pneumonia. Specifically, atypical pneumonia is a syndrome resulting from a relatively common group of pathogens including Chlamydophila sp., and Mycoplasma pneumoniae. The incidence of M. pneumoniae pneumonia in adults is less than the burden experienced by children. Transmission rates among families indicate children may act as a reservoir and maintain contagiousness over a long period of time ranging from months to years. In adults, M. pneumoniae typically produces a mild, "walking" pneumonia and is considered to be one of the causes of persistent cough in patients. M. pneumoniae has also been shown to trigger the exacerbation of other lung diseases. It has been repeatedly detected in patients with bronchitis, asthma, chronic obstructive pulmonary disorder, and cystic fibrosis. Recent advances in technology allow for the rapid diagnosis of M. pneumoniae through the use of polymerase chain reaction or rapid antigen tests. With this, more effort has been afforded to identify the causative etiologic agent in all cases of pneumonia. However, previous practices, including the overprescribing of macrolide treatment in China and Japan, have created increased incidence of macrolide-resistant M. pneumoniae. Reports from these countries indicate that >85% of M. pneumoniae pneumonia pediatric cases are macrolide-resistant. Despite its extensively studied past, the smallest bacterial species still inspires some of the largest questions. The developments in microbiology, diagnostic features and techniques, epidemiology, treatment and vaccines, and upper respiratory conditions associated with M. pneumoniae in adult populations are included within this review.

  8. Mycoplasma genitalium: An emergent sexually transmitted disease?

    PubMed

    Manhart, Lisa E

    2013-12-01

    This article summarizes the epidemiologic evidence linking Mycoplasma genitalium to sexually transmitted disease syndromes, including male urethritis, and female cervicitis, pelvic inflammatory disease, infertility, and adverse birth outcomes. It discusses the relationship of this bacterium to human immunodeficiency virus infection and reviews the available literature on the efficacy of standard antimicrobial therapies against M genitalium.

  9. Mycoplasma Pneumoniae Infections of Adults and Children

    PubMed Central

    Cherry, James D.; Welliver, Robert C.

    1976-01-01

    Although the hallmark of Mycoplasma pneumoniae infection is pneumonia, the organism is also responsible for a protean array of other symptoms. With an increased awareness of the board clinical spectrum of M. pneumoniae disease and the ready availability of the cold agglutinin and M. pneumoniae complement-fixation tests, interested clinicians will note additional clinical-mycoplasmal associations in their patients. PMID:782043

  10. Mycoplasma wenyonii infection in dairy cows.

    PubMed

    2016-12-17

    Mycoplasma wenyonii infection in two dairy herdsHypomagnesaemia in suckler cowsLeptospiral milk drop in dairy cowsNon-resolving orf in six-month-old lambsHepatosis dietetica in a five-month-old giltThese are among matters discussed in the disease surveillance report for September 2016 from SAC Consulting: Veterinary Services (SAC C VS). British Veterinary Association.

  11. Isolation and characterization of unusual Mycoplasma spp. from captive Eurasian Griffon (Gyps fulvus) in Sicily.

    PubMed

    Loria, G R; Ferrantelli, E; Giardina, G; Li Vecchi, L; Sparacino, L; Oliveri, F; McAuliffe, L; Nicholas, R A J

    2008-01-01

    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.

  12. Development of fluorescence expression tools to study host-mycoplasma interactions and validation in two distant mycoplasma clades.

    PubMed

    Bonnefois, Tiffany; Vernerey, Marie-Stéphanie; Rodrigues, Valérie; Totté, Philippe; Puech, Carinne; Ripoll, Chantal; Thiaucourt, François; Manso-Silván, Lucía

    2016-10-20

    Fluorescence expression tools for stable and innocuous whole mycoplasma cell labelling have been developed. A Tn4001-derivative mini-transposon affording unmarked, stable mutagenesis in mycoplasmas was modified to allow the constitutive, high-level expression of mCherry, mKO2 and mNeonGreen. These tools were used to introduce the respective fluorescent proteins as chromosomal tags in the phylogenetically distant species Mycoplasma mycoides subsp. mycoides and Mycoplasma bovis. The production, selection and characterisation of fluorescent clones were straightforward and resulted in the unprecedented observation of red and green fluorescent mycoplasma colonies in the two species, with no apparent cytotoxicity. Equivalent fluorescence expression levels were quantified by flow cytometry in both species, suggesting that these tools can be broadly applied in mycoplasmas. A macrophage infection assay was performed to assess the usefulness of mNeonGreen-expressing strains for monitoring mycoplasma infections, and notably cell invasion. The presence of fluorescent mycoplasmas inside live phagocytic cells was detected and quantified by flow cytometry and corroborated by confocal microscopy, which allowed the identification of individual mycoplasmas in the cytoplasm of infected cells. The fluorescence expression tools developed in this study are suitable for host-pathogen interaction studies and offer innumerable perspectives for the functional analysis of mycoplasmas both in vitro and in vivo.

  13. Postoperative Mycoplasma hominis infections after neurosurgical intervention.

    PubMed

    Whitson, Wesley J; Ball, Perry A; Lollis, S Scott; Balkman, Jason D; Bauer, David F

    2014-08-01

    Mycoplasma hominis is a rare cause of infection after neurosurgical procedures. The Mycoplasma genus contains the smallest bacteria discovered to date. Mycoplasma are atypical bacteria that lack a cell wall, a feature that complicates both diagnosis and treatment. The Gram stain and some types of culture media fail to identify these organisms, and typical broad-spectrum antibiotic regimens are ineffective because they act on cell wall metabolism. Mycoplasma hominis commonly colonizes the genitourinary tract in a nonvirulent manner, but it has caused postoperative, postpartum, and posttraumatic infections in various organ systems. The authors present the case of a 17-year-old male with a postoperative intramedullary spinal cord abscess due to M. hominis and report the results of a literature review of M. hominis infections after neurosurgical procedures. Attention is given to time to diagnosis, risk factors for infection, ineffective antibiotic regimens, and final effective antibiotic regimens to provide pertinent information for the practicing neurosurgeon to diagnose and treat this rare occurrence. A PubMed search was performed to identify reports of M. hominis infections after neurosurgical procedures. Eleven cases of postneurosurgical M. hominis infection were found. No other cases of intramedullary spinal cord abscess were found. Initial antibiotic coverage was inadequate in all cases, and diagnosis was delayed in all cases. Multiple surgical interventions were often needed. Once appropriate antibiotics were started, patients typically experienced rapid resolution of their neurological symptoms. In 27% of cases, a suspicious genitourinary source other than urinary catheterization was identified. Postoperative M. hominis infections are rarely seen after neurosurgical procedures. They are typically responsive to appropriate antibiotic therapy. Mycoplasma infection may cause prolonged hospitalization and multiple returns to the operating room due to delay in

  14. Transcriptome changes in Mycoplasma hyopneumoniae during infection.

    PubMed

    Madsen, Melissa L; Puttamreddy, Supraja; Thacker, Eileen L; Carruthers, Michael D; Minion, F Chris

    2008-02-01

    Mycoplasma hyopneumoniae causes swine pneumonia and contributes significantly to the porcine respiratory disease complex. The mechanisms of pathogenesis are difficult to address, since there is a lack of genetic tools, but microarrays are available and can be used to study transcriptional changes that occur during disease as a way to identify important virulence-related genes. Mycoplasmas were collected from bronchial alveolar lavage samples and compared to broth-grown cells using microarrays. Bronchial alveolar lavage was performed on pigs 28 days postinfection, and mycoplasmas were isolated by differential centrifugation. Mycoplasma RNA-enriched preparations were then obtained from total RNA by subtracting eucaryotic ribosomal and messenger RNAs. Labeled cDNAs were generated with mycoplasma open reading frame-specific primers. Nine biological replicates were analyzed. During lung infection, our analysis indicated that 79 M. hyopneumoniae genes were differentially expressed (P < 0.01), at a false-discovery rate of <2.7%. Of the down-regulated genes, 28 of 46 (61%) lacked an assigned function, in comparison to 21 of 33 (63%) of up-regulated genes. Four down-regulated genes and two up-regulated genes encoded putative lipoproteins. secA (mhp295) (P = 0.003) and two glycerol transport permease genes (potA [mhp380; P = 0.006] and ugpA [mhp381; P = 0.003]) were up-regulated in vivo. Elongation factor EF-G (fusA [mhp083]) (P = 0.002), RNA polymerase beta chain (rpoC [mhp635]) (P = 0.003), adenylate kinase (adk [mhp208]) (P = 0.001), prolyl aminoacyl tRNA synthetase (proS [mhp397]) (P = 0.009), and cysteinyl-tRNA synthetase (cysS [mhp661]) (P < 0.001) were down-regulated in vivo.

  15. Antimicrobial susceptibilities of Mycoplasma isolated from bovine mastitis in Japan.

    PubMed

    Kawai, Kazuhiro; Higuchi, Hidetoshi; Iwano, Hidetomo; Iwakuma, Akihiro; Onda, Ken; Sato, Reiichiro; Hayashi, Tomohito; Nagahata, Hajime; Oshida, Toshio

    2014-01-01

    Mycoplasma spp. are highly contagious pathogens and intramammary Mycoplasma infection is a serious issue for the dairy industry. As there is no effective vaccine for Mycoplasma infection, control depends on good husbandry and chemo-antibiotic therapy. In this study, antimicrobial susceptibility of Mycoplasma strains recently isolated from cases of bovine mastitis in Japan was evaluated by minimum inhibitory concentration (MIC). All Mycoplasma bovis strains were sensitive to pirlimycin, danofloxacin and enrofloxacin, but not kanamycin, oxytetracycline, tilmicosin or tylosin. M. californicum and M. bovigenitalium strains were sensitive to pirlimycin, danofloxacin, enrofloxacin, oxytetracycline, tilmicosin and tylosin, but not to kanamycin. This is the first report to describe the MIC of major antimicrobial agents for Mycoplasma species isolated from bovine mastitis in Japan.

  16. Conjunctivitis, rhinitis, and sinusitis in cliff swallows (Petrochelidon pyrrhonota) found in association with Mycoplasma sturni infection and cryptosporidiosis.

    PubMed

    Ley, David H; Moresco, Anneke; Frasca, Salvatore

    2012-01-01

    Fledgling cliff swallows were cared for at a rehabilitation facility when clinical signs of ocular disease, characterized by conjunctivitis, epiphora, and hyperaemia of palpebrae and nictitans, were recognized. Treatment consisted of topical and oral antibiotic therapy and one topical steroid administration. However, one cliff swallow died and three were killed due to poor therapeutic response. Conjunctival swabs were obtained ante-mortem from the three cliff swallows and were submitted for mycoplasma culture and molecular diagnostics. Heads of the three birds were fixed in 10% neutral buffered formalin and submitted for histopathologic examination of oculonasal tissues. Mycoplasma cultures and molecular evaluation of isolates identified Mycoplasma sturni, but not Mycoplasma gallisepticum, from each specimen. Histopathologic examination revealed lymphoplasmacytic conjunctivitis, rhinitis and infraorbital sinusitis with follicular lymphoid hyperplasia, epithelial hyperplasia, and protozoal stages compatible with Cryptosporidium spp. arranged in and along the apical surfaces of epithelial cells. Identification of concurrent M. sturni and Cryptosporidium spp. infections in these cliff swallows demonstrates an alternative infectious condition that can produce gross and microscopic lesions comparable with those commonly observed in M. gallisepticum infections of house finches and other passerine species. Conjunctivitis associated with M. sturni and Cryptosporidium spp. in cliff swallows may represent an emerging disease risk to a naïve, high-density and colonial species such as colony-nesting cliff swallows.

  17. [Sensorineural hearing impairment in combination with mycoplasma infection].

    PubMed

    Gurov, A V; Levina, Yu V; Rudenko, V V

    2015-01-01

    The objective of the present study was to elucidate the incidence of mycoplasma infection concomitant with sensorineural hearing impairment and its clinical manifestations with special reference to the methods for its diagnostics and treatment. The main method for the detection of mycoplasma infection is PCR in real time and the auxiliary one is the immunoenzymatic assay. The study revealed mycoplasma infection in 15 (13.9%) of the examined patients. The results of our investigations give evidence of the necessity to further study the clinical symptoms of mycoplasma infection associated with sensorineural hearing impairment and to search for the methods of the management of this condition.

  18. Secretomes of Mycoplasma hyopneumoniae and Mycoplasma flocculare reveal differences associated to pathogenesis.

    PubMed

    Paes, Jéssica A; Lorenzatto, Karina R; de Moraes, Sofia N; Moura, Hercules; Barr, John R; Ferreira, Henrique B

    2017-02-10

    Mycoplasma hyopneumoniae and Mycoplasma flocculare cohabit the porcine respiratory tract. However, M. hyopneumoniae causes the porcine enzootic pneumonia, while M. flocculare is a commensal bacterium. Comparative analyses demonstrated high similarity between these species, which includes the sharing of all predicted virulence factors. Nevertheless, studies related to soluble secretomes of mycoplasmas were little known, although they are important for bacterial-host interactions. The aim of this study was to perform a comparative analysis between the soluble secreted proteins repertoires of the pathogenic Mycoplasma hyopneumoniae and its closely related commensal Mycoplasma flocculare. For that, bacteria were cultured in medium with reduced serum concentration and secreted proteins were identified by a LC-MS/MS proteomics approach. Altogether, 62 and 26 proteins were identified as secreted by M. hyopneumoniae and M. flocculare, respectively, being just seven proteins shared between these bacteria. In M. hyopneumoniae secretome, 15 proteins described as virulence factors were found; while four putative virulence factors were identified in M. flocculare secretome. For the first time, clear differences related to virulence were found between these species, helping to elucidate the pathogenic nature of M. hyopneumoniae to swine hosts.

  19. Synergism between upregulation of Rab7 and inhibition of autophagic degradation caused by mycoplasma facilitates intracellular mycoplasma infection.

    PubMed

    Hu, Xiaopeng; Yu, Jie; Zhou, Xiang; Li, Zhaoming; Xia, Yun; Luo, Zhiyong; Wu, Yaqun

    2014-03-01

    Following fusion of a mycoplasma with a host cell membrane, the inserted components of mycoplasma may then be transported through the endocytic pathway. However, the effects of mycoplasmas on the host cell endomembrane system are largely unknown. In this study, mycoplasma‑induced changes in the dynamics of endocytic and autophagic systems were investigated. Endocytosis and autophagy are two major processes involved in the survival of intracellular prokaryotic pathogens. It was found that, immediately following infection, mycoplasmas induce endocytosis in the host cell; however, in the long term the mycoplasmas suppress turnover of the components of the endocytic pathway. Immunofluorescence microscopy revealed that Rab7 and LC3‑II are recruited to the intracellular mycoplasma‑containing compartments. Western blot analysis and quantitative reverse transcription-polymerase chain reaction (qPCR) showed that mycoplasmas increase expression of Rab7 by upregulating transcription, but increase levels of LC3‑II and p62 by post‑translational regulation. Furthermore, it was demonstrated that mycoplasma infection causes inhibition of autophagic degradation of LC3‑II and p62. In addition, it was found that upregulation of Rab7 and inhibition of autophagic degradation synergistically contributes to intracellular mycoplasma accumulation. In conclusion, these findings suggest that mycoplasmas may manipulate host cell endosomal and autophagic systems in order to facilitate intracellular infection.

  20. [Refinement and study of the performance of a "Mycoplasma UG" kit for the research on urogenital Mycoplasmas].

    PubMed

    Melzi, A; Atieh, S; Benyahia, H

    1998-01-01

    The pathogenic role of genital Mycoplasmas is no more disputably. The diagnosis is based on the culture which may present some technical difficulties. The present study consist in a perfecting of a miniaturised system called "Mycoplasma UG" for the research, identification and titration of genital Mycoplasmas. The performance of the kit is studied with an important number of different types samples by comparison with the commercial kit "Mycoplasma Duo" and the classic reference method. The results obtained are satisfactory and constitute an important element in favour of a more advanced validation before the kit's trading.

  1. [Localization of the division protein FtsZ in mycoplasma cells Mycoplasma hominis].

    PubMed

    Vishniakov, I E; Borkhsenius, S N; Basovskiĭ, Iu I; Levitskiĭ, S A; Lazarev, V N; Snigirevskaia, E S; Komissarchik, Ia Iu

    2009-01-01

    Localization of the protein FtsZ in Mycoplasma hominis cells was determined. Ultra thin sections were treated by rabbit polyclonal antibodies against FtsZ M. hominis: a conjugate of protein A with colloidal gold particles was used instead of secondary antibodies. Considerable polymorphism of cells was seen on electron microscopy pictures of M. hominis cells, which is typical for mycoplasmas. Among a wide variety of cell shapes we distinguished dumbbell-shaped dividing cells, and the cells connected with each other with the aid of thin membrane tubules (former constrictions). Dominants distribution of the label in the constriction area of dividing M. hominis cells and in the area of the thin membrane tubules was observed. We revealed the cross septum in the mycoplasma cells for the first time, as well as the gold labeling of this structure. Furthermore, in some rounded and oval cells colloidal gold particles labeled the whole plasma membrane in ring-shaped manner. Probably, the label in these cases marks a submembrane contractile ring (Z-ring). The facts mentioned above confirm that FtsZ of M. hominis plays an active role in the mycoplasma cytokinesis. In a series of cases spiral-like distribution of gold particles was observed. Probably, FtsZ protofilaments in M. hominis cells can form spiral structures similar to Z-spirals of Bacillus subtilis and Escherichia coli. Its presence in mycoplasma cells may be considered as an important argument in favour of model of Z-ring assembling through reorganization of Z-spirals. FtsZ also may participate in maintenance of mycoplasma cell shape (membrane localization).

  2. Development of Mycoplasma hyopneumoniae Recombinant Vaccines.

    PubMed

    Marchioro, Silvana Beutinger; Simionatto, Simone; Dellagostin, Odir

    2016-01-01

    Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia (EP), a disease that affects swine production worldwide. Vaccination is the most cost-effective strategy for the control and prevention of the disease. Research using genome-based approach has the potential to elucidate the biology and pathogenesis of M. hyopneumoniae and contribute to the development of more effective vaccines. Here, we describe the protocol for developing M. hyopneumoniae recombinant vaccines using reverse vaccinology approaches.

  3. Mycoplasma bovis arthritis and pneumonia in calves.

    PubMed

    2017-03-18

    Mycoplasma bovis arthritis and pneumonia in six-month-old calvesSudden deaths in housed suckler cows due to hypomagnesaemiaBovine respiratory syncytial virus infection in two-year-old heifersBovine abortion associated with Parachlamydia speciesFibrinous pericarditis due to Aeromonas hydrophila in weaner pigsThese are among matters discussed in the disease surveillance report for December 2016 from SAC Consulting: Veterinary Services (SAC C VS). British Veterinary Association.

  4. Simultaneous Detection of Mycoplasma pneumoniae, Mycoplasma hominis and Mycoplasma arthritidis in Synovial Fluid of Patients with Rheumatoid Arthritis by Multiplex PCR.

    PubMed

    Ataee, Ramezan Ali; Golmohammadi, Reza; Alishiri, Gholam Hossein; Mirnejad, Reza; Najafi, Ali; Esmaeili, Davood; Jonaidi-Jafari, Nematollah

    2015-06-01

    It has been recognized that infectious agents, such as different bacteria and viruses, may play a role in the developing of rheumatoid arthritis (RA). Recently, the mycoplasma species has been implicated in the pathogenesis of RA. The aim of this study was to design a multiplex PCR for rapid and simultaneous detection of Mycoplasma pneumoniae, Mycoplasma hominis, and Mycoplasma arthritidis in the synovial fluid of patients with rheumatoid arthritis (RA). A total of 131 synovial fluid (SF) samples from patients with RA were assayed. Mycoplasma pneumoniae (ATCC: 29342), M. hominis (native strain), and the synthetic complete genome of M. arthritidis mitogen (MAM) superantigen were used as controls. All SF samples were subjected to DNA extraction separately and multiplex PCR was performed. The PCR products were confirmed by sequencing. The designed multiplex PCR was able to detect M. pneumoniae, M. hominis, and M. arthritidis in the SF of patients with RA with a frequency of 30 (22.9%), 23 (17.5%) and 13 (9.9%), respectively. In this study, the overall detection of the Mycoplasma species in RA patients was 53.4%; thus, we recommend the application of multiplex PCR assays when searching for a specific anti mycoplasma treatment for RA patients.

  5. Mycoplasma genitalium: from Chrysalis to Multicolored Butterfly

    PubMed Central

    Taylor-Robinson, David; Jensen, Jørgen Skov

    2011-01-01

    Summary: The history, replication, genetics, characteristics (both biological and physical), and factors involved in the pathogenesis of Mycoplasma genitalium are presented. The latter factors include adhesion, the influence of hormones, motility, possible toxin production, and immunological responses. The preferred site of colonization, together with current detection procedures, mainly by PCR technology, is discussed. The relationships between M. genitalium and various diseases are highlighted. These diseases include acute and chronic nongonococcal urethritis, balanoposthitis, chronic prostatitis, and acute epididymitis in men and urethritis, bacterial vaginosis, vaginitis, cervicitis, pelvic inflammatory disease, and reproductive disease in women. A causative relationship, or otherwise strong association, between several of these diseases and M. genitalium is apparent, and the extent of this, on a subjective basis, is presented; also provided is a comparison between M. genitalium and two other genital tract-orientated mollicutes, namely, Mycoplasma hominis, the first mycoplasma of human origin to be discovered, and Ureaplasma species. Also discussed is the relationship between M. genitalium and infertility and also arthritis in both men and women, as is infection in homosexual and immunodeficient patients. Decreased immunity, as in HIV infections, may enhance mycoplasmal detection and increase disease severity. Finally, aspects of the antimicrobial susceptibility and resistance of M. genitalium, together with the treatment and possible prevention of mycoplasmal disease, are discussed. PMID:21734246

  6. Characterization of Canine Mycoplasmas by Polyacrylamide Gel Electrophoresis and Immunodiffusion

    PubMed Central

    Armstrong, D.; Yu, B.

    1970-01-01

    Canine mycoplasmas which had been characterized by biological and serological methods were further studied by using polyacrylamide gel electrophoresis (PGE) and double diffusion in agar gel. The three dog mycoplasmas previously characterized, Mycoplasma canis, M. maculosum, and M. spumans showed distinctive patterns by PGE. Five additional representative isolates from dogs had been characterized serologically and biologically into three new groups, A, C, and D. An additional mycoplasma (group B) was indistinguishable from M. canis by growth inhibition and PGE but was more broadly reactive with field isolates serologically. The group A organisms were distinctive in pattern and similar to those studied by Razin and Rottem, tentatively designated M. edwardii. The group C organisms were represented by two isolates which were similar by fluorescent-antibody studies but different by growth inhibition tests. These two isolates were also different from each other by PGE. The group D serotypes were also distinctive by PGE from all other dog mycoplasmas tested. It was found, during these studies, that two different mycoplasmas showed different PGE patterns at different intervals during incubation. Immunodiffusion studies showed a relationship among all the canine mycoplasmas, and bands of nonidentity between the two group C mycoplasmas were demonstrated. Images PMID:4990761

  7. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... infection to mild or severe upper respiratory disease, ear infection, and bronchial pneumonia. (b.... The identification aids in the diagnosis of disease caused by bacteria belonging to the genus Mycoplasma and provides epidemiological information on diseases caused by these microorganisms. Mycoplasma...

  8. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... infection to mild or severe upper respiratory disease, ear infection, and bronchial pneumonia. (b.... The identification aids in the diagnosis of disease caused by bacteria belonging to the genus Mycoplasma and provides epidemiological information on diseases caused by these microorganisms. Mycoplasma...

  9. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... infection to mild or severe upper respiratory disease, ear infection, and bronchial pneumonia. (b.... The identification aids in the diagnosis of disease caused by bacteria belonging to the genus Mycoplasma and provides epidemiological information on diseases caused by these microorganisms. Mycoplasma...

  10. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... infection to mild or severe upper respiratory disease, ear infection, and bronchial pneumonia. (b.... The identification aids in the diagnosis of disease caused by bacteria belonging to the genus Mycoplasma and provides epidemiological information on diseases caused by these microorganisms. Mycoplasma...

  11. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... infection to mild or severe upper respiratory disease, ear infection, and bronchial pneumonia. (b.... The identification aids in the diagnosis of disease caused by bacteria belonging to the genus Mycoplasma and provides epidemiological information on diseases caused by these microorganisms. Mycoplasma...

  12. Mycoplasma hominis, a Rare but True Cause of Infective Endocarditis.

    PubMed

    Gagneux-Brunon, Amandine; Grattard, Florence; Morel, Jerome; Suy, Florence; Fuzellier, Jean-François; Verhoeven, Paul; Cazorla, Celine; Guglielminotti, Claire; Fresard, Anne; Lucht, Frederic; Botelho-Nevers, Elisabeth

    2015-09-01

    Mycoplasma spp. are rarely recognized agents of infective endocarditis. We report a case of Mycoplasma hominis prosthetic valve endocarditis diagnosed by 16S ribosomal DNA (rDNA) PCR and culture of valves in a 74-year-old man. We reviewed the literature and found only 8 other cases reported.

  13. "Candidatus Mycoplasma haemomacaque" and Bartonella quintana bacteremia in cynomolgus monkeys.

    PubMed

    Maggi, Ricardo G; Mascarelli, Patricia E; Balakrishnan, Nandhakumar; Rohde, Cynthia M; Kelly, Catherine M; Ramaiah, Lila; Leach, Michael W; Breitschwerdt, Edward B

    2013-05-01

    Here, we report latent infections with Bartonella quintana and a hemotropic Mycoplasma sp. in a research colony of cynomolgus monkeys (Macaca fascicularis). Sequence alignments, evolutionary analysis, and signature nucleotide sequence motifs of the hemotropic Mycoplasma 16S rRNA and RNase P genes indicate the presence of a novel organism.

  14. Mycoplasma hominis, a Rare but True Cause of Infective Endocarditis

    PubMed Central

    Grattard, Florence; Morel, Jerome; Suy, Florence; Fuzellier, Jean-François; Verhoeven, Paul; Cazorla, Celine; Guglielminotti, Claire; Fresard, Anne; Lucht, Frederic; Botelho-Nevers, Elisabeth

    2015-01-01

    Mycoplasma spp. are rarely recognized agents of infective endocarditis. We report a case of Mycoplasma hominis prosthetic valve endocarditis diagnosed by 16S ribosomal DNA (rDNA) PCR and culture of valves in a 74-year-old man. We reviewed the literature and found only 8 other cases reported. PMID:26135868

  15. In vitro susceptibilities of Mycoplasma genitalium to antibiotics.

    PubMed Central

    Renaudin, H; Tully, J G; Bebear, C

    1992-01-01

    The susceptibilities of seven clinical isolates of Mycoplasma genitalium and three strains of Mycoplasma pneumoniae to a variety of antibiotics were examined by an agar dilution method. Macrolides, pristinamycin, and tetracyclines were very active against both species. Sparfloxacin was the most active quinolone tested. None of the 21 antibiotics tested had differential activity toward the two organisms. PMID:1503451

  16. [Role of mycoplasmas in urogenital tract infections and their complications].

    PubMed

    Kłosowska, Wioletta Marta; Zdrodowska-Stefanow, Bozena; Wilkowska-Trojniel, Marta

    2005-02-01

    In the literature from recent years it is stressed the significant epidemiological and clinical role of the sexually transmitted infections caused by so called "new generation" pathogens including mycoplasmas. The paper gives a review of the current literature concerning the epidemiology, clinical presentation, diagnosis and therapy of the infections of urogenital tract caused by mycoplasmas.

  17. 9 CFR 113.28 - Detection of mycoplasma contamination.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Detection of mycoplasma contamination. 113.28 Section 113.28 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE... mycoplasma medium. In the case of a cell line or a sample of primary cells, the inoculum shall consist of the...

  18. 9 CFR 113.28 - Detection of mycoplasma contamination.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Detection of mycoplasma contamination. 113.28 Section 113.28 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE... mycoplasma medium. In the case of a cell line or a sample of primary cells, the inoculum shall consist of the...

  19. Alice in Wonderland syndrome associated with mycoplasma infection.

    PubMed

    Omata, Taku; Fujii, Katsunori; Kuroki, Haruo; Shimojo, Naoki

    2016-10-01

    Alice in Wonderland syndrome (AIWS) is a rare condition in which patients report distorted size perception of objects and their own bodies. Although specific causes and pathology have not been elucidated, an association between AIWS and infection has been suggested. To our knowledge, mycoplasma-induced AIWS has not been examined. A girl aged 7 years 11 months presented with fever (temperature, 40°C) and cough. Although the fever disappeared after approximately 10 days, she complained that her mother's face suddenly appeared smaller to her. Subsequently, she complained that objects intermittently appeared smaller than normal. Particle agglutination test indicated elevated serum antibodies against Mycoplasma pneumoniae. The patient was therefore diagnosed the patient with AIWS secondary to mycoplasma infection. Although mycoplasma infection is known to cause various central nervous system symptoms, this is the first report involving AIWS, suggesting that mycoplasma could affect visual function in children.

  20. Putative essential and core-essential genes in Mycoplasma genomes.

    PubMed

    Lin, Yan; Zhang, Randy Ren

    2011-01-01

    Mycoplasma, which was used to create the first "synthetic life", has been an important species in the emerging field, synthetic biology. However, essential genes, an important concept of synthetic biology, for both M. mycoides and M. capricolum, as well as 14 other Mycoplasma with available genomes, are still unknown. We have developed a gene essentiality prediction algorithm that incorporates information of biased gene strand distribution, homologous search and codon adaptation index. The algorithm, which achieved an accuracy of 80.8% and 78.9% in self-consistence and cross-validation tests, respectively, predicted 5880 essential genes in the 16 Mycoplasma genomes. The intersection set of essential genes in available Mycoplasma genomes consists of 153 core essential genes. The predicted essential genes (available from pDEG, tubic.tju.edu.cn/pdeg) and the proposed algorithm can be helpful for studying minimal Mycoplasma genomes as well as essential genes in other genomes.

  1. Putative essential and core-essential genes in Mycoplasma genomes

    PubMed Central

    Lin, Yan; Zhang, Randy Ren

    2011-01-01

    Mycoplasma, which was used to create the first “synthetic life”, has been an important species in the emerging field, synthetic biology. However, essential genes, an important concept of synthetic biology, for both M. mycoides and M. capricolum, as well as 14 other Mycoplasma with available genomes, are still unknown. We have developed a gene essentiality prediction algorithm that incorporates information of biased gene strand distribution, homologous search and codon adaptation index. The algorithm, which achieved an accuracy of 80.8% and 78.9% in self-consistence and cross-validation tests, respectively, predicted 5880 essential genes in the 16 Mycoplasma genomes. The intersection set of essential genes in available Mycoplasma genomes consists of 153 core essential genes. The predicted essential genes (available from pDEG, tubic.tju.edu.cn/pdeg) and the proposed algorithm can be helpful for studying minimal Mycoplasma genomes as well as essential genes in other genomes. PMID:22355572

  2. Significance of Genital Mycoplasmas in Pelvic Inflammatory Disease: Innocent Bystander!

    PubMed Central

    Harmanli, Ozgur H.; Nyirjesy, Paul; Reece, E. Albert

    1996-01-01

    Objective: Our objective was to determine the role of Mycoplasma hominis and Ureaplasma urealyticum in pelvic inflammatory disease (PID). Methods: The clinical and microbiologic variables in 114 patients with a clinical diagnosis of PID were compared prospectively according to the isolation of M. hominis and U. urealyticum from their endometrial cavities. Results: The groups were epidemiologically well matched. Clinical parameters such as temperature, leukocyte count, erythrocyte count, and C-reactive protein on admission and length of hospital stay were similar in the patients, regardless of their mycoplasma status. A significant percentage of the patients either continued or started to harbor genital mycoplasmas after the resolution of PID without any significant clinical sequelae. Conclusions: The presence of genital mycoplasmas does not change the clinical presentation and course of PID. Both M. hominis and U. urealyticum can persist or colonize the endometrium after complete recovery from PID. Therefore, the genital mycoplasmas do not seem to have a dominant pathogenic role in PID. PMID:18476105

  3. Use of Real-Time PCR To Detect and Quantify Mycoplasma haemofelis and “Candidatus Mycoplasma haemominutum” DNA

    PubMed Central

    Tasker, Séverine; Helps, Chris R.; Day, Michael J.; Gruffydd-Jones, Tim J.; Harbour, Dave A.

    2003-01-01

    A real-time PCR assay using Taqman probes was developed to detect and quantify Mycoplasma haemofelis and “Candidatus Mycoplasma haemominutum” in feline blood samples. The assay was rapid and sensitive and was successfully used to monitor the in vivo kinetics of cats experimentally infected with each species. PMID:12517888

  4. Hemotropic mycoplasmas in little brown bats (Myotis lucifugus).

    PubMed

    Mascarelli, Patricia E; Keel, Michael K; Yabsley, Michael; Last, Lisa A; Breitschwerdt, Edward B; Maggi, Ricardo G

    2014-03-24

    Hemotropic mycoplasmas are epicellular erythrocytic bacteria that can cause infectious anemia in some mammalian species. Worldwide, hemotropic mycoplasmas are emerging or re-emerging zoonotic pathogens potentially causing serious and significant health problems in wildlife. The objective of this study was to determine the molecular prevalence of hemotropic Mycoplasma species in little brown bats (Myotis lucifugus) with and without Pseudogymnoascus (Geomyces) destrucans, the causative agent of white nose syndrome (WNS) that causes significant mortality events in bats. In order to establish the prevalence of hemotropic Mycoplasma species in a population of 68 little brown bats (Myotis lucifugus) with (n = 53) and without (n = 15) white-nose syndrome (WNS), PCR was performed targeting the 16S rRNA gene. The overall prevalence of hemotropic Mycoplasmas in bats was 47%, with similar (p = 0.5725) prevalence between bats with WNS (49%) and without WNS (40%). 16S rDNA sequence analysis (~1,200 bp) supports the presence of a novel hemotropic Mycoplasma species with 91.75% sequence homology with Mycoplasma haemomuris. No differences were found in gene sequences generated from WNS and non-WNS animals. Gene sequences generated from WNS and non-WNS animals suggest that little brown bats could serve as a natural reservoir for this potentially novel Mycoplasma species. Currently, there is minimal information about the prevalence, host-specificity, or the route of transmission of hemotropic Mycoplasma spp. among bats. Finally, the potential role of hemotropic Mycoplasma spp. as co-factors in the development of disease manifestations in bats, including WNS in Myotis lucifugus, remains to be elucidated.

  5. Mollicutes in vaginal microbiology: Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum and Mycoplasma genitalium.

    PubMed

    Taylor-Robinson, David

    2017-03-02

    Mycoplasma hominis was isolated in 1937 from the human genital tract, followed 17 years later by Ureaplasma urealyticum and 27 years after that by Mycoplasma genitalium. The first two proved relatively easy to culture but the latter required a polymerase chain reaction assay for further studies. In sexually mature women, M. hominis may be found in the vagina/cervix of about 20-50%, ureaplasmas in 40-80% and M. genitalium in 0-5%. Some heterogeneity has been found among strains of all these species, sufficient to divide ureaplasmas into two species, namely U. urealyticum and Ureaplasma parvum. Studies in female mice show that sex hormones have a profound influence on colonization, multiplication and persistence of mycoplasmas/ureaplasmas in the genital tract and provoke the question, unanswered, of whether there is such an effect in the human tract. In women, there is no evidence that any of the mycoplasmal species stimulate an inflammatory vaginitis. M. hominis organisms increase hugely in number in the case of bacterial vaginosis (BV), and to a lesser extent so do ureaplasmas. Despite this, they have not been incriminated as a sole cause of BV. Evidence for the involvement of M. genitalium remains controversial. The strong association of BV with preterm birth raises the possibility that the genital mycoplasmas might play a part, but assurance that any do will be difficult to obtain. Detailed examination of the vaginal microbiome has not yet provided an answer. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  6. Breeder turkey hens seropositive and culture-negative for Mycoplasma synoviae.

    PubMed

    Avakian, A P; Ley, D H; Berkhoff, J E; Ficken, M D

    1992-01-01

    Four flocks of clinically normal turkey breeder hens were shown to have suspect and positive Mycoplasma synoviae (MS) hemagglutination-inhibition (HI), enzyme-linked immunosorbent assay, and, in some cases, serum plate agglutination serology in the absence of MS isolation. In all cases, HI serology for Mycoplasma gallisepticum (MG) and M. meleagridis was negative. Acholeplasma laidlawii was isolated from some hens in each of these MS-seropositive culture-negative flocks. Immunoblotting was used to help determine if this positive MS serology was a result of cross-reactive antibodies to A. laidlawii or to some other Mycoplasma species. When sera from two of the flocks were reacted with MS antigen in immunoblotting, a strong and characteristic MS immunoblot profile was seen. Immunoblotting gave no evidence of a strong antibody response to A. laidlawii, M. iowae, or MG. This suggests the presence (or earlier presence) of MS in these flocks that is difficult to isolate by routine methods. Furthermore, this work shows that immunoblotting can be an important tool in the diagnosis of poultry diseases.

  7. New models of chronic synovitis in rabbits induced by mycoplasmas: microbiological, histopathological, and immunological observations on rabbits injected with Mycoplasma arthritidis and Mycoplasma pulmonis.

    PubMed Central

    Cole, B C; Griffiths, M M; Eichwald, E J; Ward, J R

    1977-01-01

    A dose-dependent chronic synovitis was induced in rabbit knees after the intra-articular injection of both Mycoplasma arthritidis and Mycoplasma pulmonis. The inflammation progressed from an initial acute phase at 1 week characterized by edema, infiltration of the synovium with monocytes and heterophils, and desquamation of lining cells, to a more chronic phase at 1 and 3 months, in which villus hyperplasia, lymph "nodules," mononuclear cell infiltration, fibroplasia, and collagen deposition were prominent. With one exception, mycoplasmas could no longer be cultivated from the joints 1 month postinoculation. Both mycoplasma species evoked a humoral antibody response that was more marked in synovial fluids than in peripheral blood. A cell-mediated immune reaction, as evidence by enhanced uptake by [3H]thymidine by sensitized blood, spleen, or node lymphocytes in the presence of homologous antigen, was detected only in rabbits injected with M. pulmonis. Lymphocytes taken from arthritic rabbits were no more cytotoxic toward synovial cells derived from normal or arthritic rabbits than were normal lymphocytes. The models of synovitis described in this study offer a convenient probe for determining the mechanisms of mycoplasma-induced inflammation, since they require only a single injection of the initiating agent and, in addition, utilize an animal host large enough for detailed investigation into the nature of mycoplasma/synovium interactions. Images PMID:873616

  8. Quantitative assessment of Mycoplasma hemadsorption activity by flow cytometry.

    PubMed

    García-Morales, Luis; González-González, Luis; Costa, Manuela; Querol, Enrique; Piñol, Jaume

    2014-01-01

    A number of adherent mycoplasmas have developed highly complex polar structures that are involved in diverse aspects of the biology of these microorganisms and play a key role as virulence factors by promoting adhesion to host cells in the first stages of infection. Attachment activity of mycoplasma cells has been traditionally investigated by determining their hemadsorption ability to red blood cells and it is a distinctive trait widely examined when characterizing the different mycoplasma species. Despite the fact that protocols to qualitatively determine the hemadsorption or hemagglutination of mycoplasmas are straightforward, current methods when investigating hemadsorption at the quantitative level are expensive and poorly reproducible. By using flow cytometry, we have developed a procedure to quantify rapidly and accurately the hemadsorption activity of mycoplasmas in the presence of SYBR Green I, a vital fluorochrome that stains nucleic acids, allowing to resolve erythrocyte and mycoplasma cells by their different size and fluorescence. This method is very reproducible and permits the kinetic analysis of the obtained data and a precise hemadsorption quantification based on standard binding parameters such as the dissociation constant K d. The procedure we developed could be easily implemented in a standardized assay to test the hemadsorption activity of the growing number of clinical isolates and mutant strains of different mycoplasma species, providing valuable data about the virulence of these microorganisms.

  9. [Female urogenital mycoplasma infection and drug sensitivity status in Changsha].

    PubMed

    Zuo, Cheng-xin; Huang, Jin-hua; Chen, Jing; Lu, Jian-yun; Xiang, Ya-ping

    2006-06-01

    To survey mycoplasma infection in female urogenital tract and analyze the drug sensitivity of mycoplasma in Changsha. Ureaplasma urealyticum (Uu) and Mycoplasma hominis (Mh) were detected in 6566 cases of female urogenital tract infection by means of mycoplasma culture and drug sensitivity reagent kit. Sensitivity tests for 10 antibiotics were also performed. A total of 2938 cases were mycoplasma-positive (positivity rate of 44.75%), including 2469 Uu-positive cases (37.6%), 52 Mh-positive cases (0.08%) and 417 cases positive for both Uu and Mh (6.35%). Josamycin, doxycycline, clarithromycin and azithromycin were more effective against Uu infection. Josamycin, doxycycline and thiamphenicol were more effective against Mh infection. Mixed infection with Uu and Mh was more resistant to most antibiotics but Josamycin and doxycycline. The female urogenital mycoplasma infection results mainly from Uu. Compared with simple Uu or Mh infection, mixed infection with Uu and Mh has significantly greater resistance to a wider variety of drugs. Josamycin and doxycycline are the primary choice for female urogenital mycoplasma infection in Changsha.

  10. Neutrophil histamine contributes to inflammation in mycoplasma pneumonia

    PubMed Central

    Xu, Xiang; Zhang, Dongji; Zhang, Hong; Wolters, Paul J.; Killeen, Nigel P.; Sullivan, Brandon M.; Locksley, Richard M.; Lowell, Clifford A.; Caughey, George H.

    2006-01-01

    Mycoplasmas cause chronic inflammation and are implicated in asthma. Mast cells defend against mycoplasma infection and worsen allergic inflammation, which is mediated partly by histamine. To address the hypothesis that mycoplasma provokes histamine release, we exposed mice to Mycoplasma pulmonis, comparing responses in wild-type and mast cell–deficient KitW-sh/KitW-sh (W-sh) mice. Low histamine levels in uninfected W-sh mice confirmed the conventional wisdom that mast cells are principal sources of airway and serum histamine. Although mycoplasma did not release histamine acutely in wild-type airways, levels rose up to 50-fold above baseline 1 week after infection in mice heavily burdened with neutrophils. Surprisingly, histamine levels also rose profoundly in infected W-sh lungs, increasing in parallel with neutrophils and declining with neutrophil depletion. Furthermore, neutrophils from infected airway were highly enriched in histamine compared with naive neutrophils. In vitro, mycoplasma directly stimulated histamine production by naive neutrophils and strongly upregulated mRNA encoding histidine decarboxylase, the rate-limiting enzyme in histamine synthesis. In vivo, treatment with antihistamines pyrilamine or cimetidine decreased lung weight and severity of pneumonia and tracheobronchitis in infected W-sh mice. These findings suggest that neutrophils, provoked by mycoplasma, greatly expand their capacity to synthesize histamine, thereby contributing to lung and airway inflammation. PMID:17158962

  11. RESPIRATORY PATHWAYS IN THE MYCOPLASMA II.

    PubMed Central

    VanDemark, P. J.; Smith, P. F.

    1964-01-01

    VanDemark, P. J. (University of South Dakota, Vermillion), and P. F. Smith. Respiratory pathways in the Mycoplasma. II. Pathway of electron transport during oxidation of reduced nicotinamide adenine dinucleotide by Mycoplasma hominis. J. Bacteriol. 88:122–129. 1964.—Unlike the flavin-terminated respiratory pathway of the fermentative Mycoplasma, the respiratory chain of the nonfermentative M. hominis strain 07 appears to be more complex, involving quinones and cytochromes in addition to flavins. In addition to reduction by reduced nicotine adenine dinucleotide (NADH) and reduced nicotine adenine dinucleotide phosphate, nonpyridine nucleotide-linked reduction of the respiratory chain of this organism occurred with succinate, lactate, and short-chained acyl coenzyme A derivatives as electron donors. Enzymes catalyzing the oxidation of NADH included an NADH oxidase, a diaphorase, a quinone reductase, and a cytochrome c reductase. The oxidation of NADH was sensitive to a variety of inhibitors, including 10−4m Atabrine, 10−3m sodium amytal, 10−5mp-chloromercuribenzoate, 10−4m antimycin A, and 10−4m potassium cyanide. The oxidase was resolved by the addition of 5% trichloroacetic acid and reactivated by the addition of flavin adenine dinucleotide but not flavin mononucleotide. The M. hominis sonic extract contained an NADH-coenzyme Q reductase. The oxidation of NADH was stimulated by the addition of either menadione or vitamin K2 (C35). The oxidase was inactivated by extraction with ether or irradiation at 360 mμ. The ether-inactivated enzyme was partially reactivated by the addition of “lipid” extract of the enzyme and coenzyme Q6. Difference spectra of the cell extracts revealed the presence of “b” and “a” type cytochromes. These cell extracts were found to contain a cyanide-and azide-sensitive cytochrome oxidase and catalase. PMID:14197876

  12. Cloning of the complete Mycoplasma pneumoniae genome.

    PubMed Central

    Wenzel, R; Herrmann, R

    1989-01-01

    The complete genome of Mycoplasma pneumoniae was cloned in an ordered library consisting of 34 overlapping or adjacent cosmids, one plasmid and two lambda phages. The genome size was determined by adding up the sizes of either the individual unique EcoRI restriction fragments of the gene bank or of the XhoI fragments of genomic M. pneumoniae DNA. The values from these calculations, 835 and 849 kbp, are in good agreement. An XhoI restriction map was constructed by identifying adjacent DNA fragments by probing with selected cosmid clones. Images PMID:2506532

  13. Laser radiation effects on Mycoplasma agalactiae

    NASA Astrophysics Data System (ADS)

    Dinu, Cerasela Z.; Grigoriu, Constantin; Dinescu, Maria; Pascale, Florentina; Popovici, Adrian; Gheorghescu, Lavinia; Cismileanu, Ana; Avram, Eugenia

    2002-08-01

    The biological effects of the laser radiation emitted by the Nd:YAG laser (second harmonic, wavelength 532 nm /fluence 32 mJ/cm2/pulse duration 6 ns) on the Mycoplasma agalactiae bacterium were studied. The radiation was found to intensify the multiplication of the bacteria irradiated in TRIS buffer (0.125 M), without however affecting the proteinic composition of the cell membrane. When the bacteria were irradiated in their growth medium (PPLO broth) being later cultivated on a solid medium (PPLO agar), the exclusive presence of the atypical colonies (granular and T-like ones) was noticed.

  14. The minimal gene complement of mycoplasma genitalium

    SciTech Connect

    Fraser, C.M.; Gocayne, J.D.; White, O.

    1995-10-20

    The complete nucleotide sequence (580,070 base pairs) of the Mycoplasma genitalium genome, the smallest known genome of any free-living organism, has been determined by whole-genome random sequencing and assembly. A total of only 470 predicted coding regions were identified that include genes required for DNA replication, transcription and translation, DNA repair, cellular transport, and energy metabolism. Comparison of this genome to that of Haemophilus influenzae suggests that differences in genome content are reflected as profound differences in physiology and metabolic capacity between these two organisms. 43 refs., 1 fig., 2 tabs.

  15. Ocular manifestations of Mycoplasma pneumoniae infection.

    PubMed

    Salzman, M B; Sood, S K; Slavin, M L; Rubin, L G

    1992-05-01

    Ocular manifestations of Mycoplasma pneumoniae infection, other than conjunctivitis, are uncommon. Optic disk swelling, optic nerve atrophy, retinal exudates and hemorrhages, and cranial nerve palsies have been infrequently reported. We describe a 15-year-old patient who developed bilateral optic disk edema and iritis during an acute infection with M. pneumoniae and review the world literature on findings associated with ocular manifestations of infection with this pathogen. Although our patient experienced complete resolution of iritis and optic disk edema after 6 weeks, several patients described in the literature have experienced permanent sequelae as a result of optic neuropathy.

  16. Neurological complications of Mycoplasma pneumoniae infection.

    PubMed

    Hely, M A; Williamson, P M; Terenty, T R

    1984-01-01

    This study documents five patients with neurological disease associated with evidence of recent Mycoplasma pneumoniae infection. Four patients had encephalitis associated with coma. Two of these had hemiparesis (one with dysphasia), one had seizures, and one had cerebellar and brainstem involvement. Two also had evidence of a radiculopathy and peripheral neuropathy. One patient had aseptic meningitis with later transverse myelitis. Three patients had multiple sites of neurological involvement. Respiratory infections preceded the neurological syndromes in four cases. Antibiotic therapy did not appear to alter the course of the disease. All patients had a favourable outcome.

  17. [Clinical score to rule out pneumonia due to Mycoplasma pneumoniae].

    PubMed

    Rodríguez de Ita, J; Torres-Quintanilla, A; Paláu-Dávila, L; Silva-Gburek, J C; Ortiz de Elguea-Lizarraga, J; Chávez Caraza, K L; Santos-Guzman, J

    2014-10-01

    The gold standard for the diagnosis of pneumonia secondary to Mycoplasma pneumoniae is the serial measurement of IgM, since an isolated test for IgM has a poor sensitivity of 31.8%. A pneumonia due to Mycoplasma pneumoniae could be of clinically different origins, thus it is possible to perform a clinical score for its early diagnosis. To develop a clinical score in order to rule out a pneumoniae secondary to Mycoplasma pneumoniae. A total of 302 patients from 0 to 18 years-old, with a diagnosis of pneumonia were evaluated and divided into two groups: Mycoplasma positive and Mycoplasma negative. Using different variables in the medical records a clinical score was calculated. Of the 302 cases studied, 34 were classified as Mycoplasma positive and 268 as Mycoplasma negative. The variables relevant to the calculation of the score were age, days with fever, and days with cough, thus providing the CAF (Cough, Age, Fever) score. Ranges were assigned for each variable and points were given for each range. A value greater than or equal to 5 meant a positive score. The CAF score was applied to the 302 cases, resulting in 164 cases of Mycoplasma positive and 138 cases of Mycoplasma negative. The CAF score had a sensitivity of 85% and specificity of 49%. The CAF score had better sensitivity than other clinical diagnostic tools. With a negative predictive value of 96% it is possible to rule out a pneumonia secondary to M. pneumoniae. The study requires a prospective study to verify the usefulness of our score. Copyright © 2013 Asociación Española de Pediatría. Published by Elsevier Espana. All rights reserved.

  18. Sequence homologies between Mycoplasma and Chlamydia spp. lead to false-positive results in chlamydial cell cultures tested for mycoplasma contamination with a commercial PCR assay.

    PubMed

    Maass, Viola; Kern, Jan Marco; Poeckl, Matthias; Maass, Matthias

    2011-10-01

    Mycoplasma contamination is a frequent problem in chlamydial cell culture. After obtaining contradictory contamination results, we compared three commercial PCR kits for mycoplasma detection. One kit signaled contamination in mycoplasma-free Chlamydia pneumoniae cultures. Sequencing of cloned PCR products revealed primer homology with the chlamydial genome as the basis of this false-positive result.

  19. Effect of a Mycoplasma hominis-like Mycoplasma on the infection of HEp-2 cells by the TW-183 strain of Chlamydia pneumoniae.

    PubMed

    Castilla, E A; Wadowsky, R M

    2000-02-01

    We isolated a Mycoplasma hominis-like mycoplasma from a stock culture of Chlamydia pneumoniae TW-183 obtained from the American Type Culture Collection and eradicated the contaminant by treating the stock suspension with a nonionic detergent, Igepal CA-630. The M. hominis-like mycoplasma neither inhibits nor enhances the infectivity of C. pneumoniae for HEp-2 cells.

  20. [Decontamination of continual cell lines spontaneously infected with mycoplasmas].

    PubMed

    Machatková, M; Jurmanová, K; Snejdar, V

    1986-07-01

    The continual cell lines of bovine kidneys MDBK and AUBEK, and porcine kidneys RPD and IBRS, spontaneously infected with Mycoplasma arginini and Acholeplasma laidlawii, were decontaminated by the method of selective elimination. Two elimination procedures were modified to be used for the decontamination: one based on the reduction of infection by the light treatment of the cultures, the other based on the selection of mycoplasma-free cell population through cell clonation. On the basis of a long-continued control of the cell clones a methodical procedure of the preparation of mycoplasma-free cell lines was worked out.

  1. The role of Mycoplasma and Ureaplasma in adverse pregnancy outcomes.

    PubMed

    Murtha, Amy P; Edwards, James M

    2014-12-01

    Genital mycoplasmas are frequently found in the vaginal flora across socioeconomic and ethnic groups and have been demonstrated to be involved in adverse perinatal outcomes. Both Mycoplasma and Ureaplasma spp cause inflammation potentially leading to spontaneous preterm birth and PPROM as well as postdelivery infectious complications and neonatal infections. Herein we have provided an overview of the existing literature and supportive evidence for genital mycoplasma's role in perinatal complications. Future research will need to focus on clearly delineating the species, allowing for discrimination of their effects.

  2. Catalase Enhances Growth and Biofilm Production of Mycoplasma pneumoniae

    PubMed Central

    Simmons, Warren L.; Dybvig, Kevin

    2015-01-01

    Mycoplasma pneumoniae causes chronic respiratory disease in humans. Factors thought to be important for colonization include the ability of the mycoplasma to form a biofilm on epithelial surfaces and the production of hydrogen peroxide to damage host tissue. Almost all of the mycoplasmas, including M. pneumoniae, lack superoxide dismutase and catalase and a balance should exist between peroxide production and growth. We show here that the addition of catalase to cultures enhanced the formation of biofilms and altered the structure. The incorporation of catalase in agar increased the number of colony-forming units detected and hence could improve the clinical diagnosis of mycoplasmal diseases. PMID:25894997

  3. Mycoplasma hominis necrotizing pleuropneumonia in a previously healthy adolescent

    PubMed Central

    2010-01-01

    Background Mycoplasma hominis is a fastidious micro-organism causing systemic infections in the neonate and genital infections in the adult. It can also be the cause of serious extra-genital infections, mainly in immunosuppressed or predisposed subjects. Case Presentation We describe a case of severe pneumonia and pericarditis due to Mycoplasma hominis in a previously healthy adolescent who did not respond to initial therapy. Conclusions Mycoplasma hominis could be an underestimated cause of severe pneumonia in immunocompetent patients and should be particularly suspected in those not responding to standard therapy. PMID:21106079

  4. Repetitive Elements in Mycoplasma hyopneumoniae Transcriptional Regulation.

    PubMed

    Cattani, Amanda Malvessi; Siqueira, Franciele Maboni; Guedes, Rafael Lucas Muniz; Schrank, Irene Silveira

    2016-01-01

    Transcriptional regulation, a multiple-step process, is still poorly understood in the important pig pathogen Mycoplasma hyopneumoniae. Basic motifs like promoters and terminators have already been described, but no other cis-regulatory elements have been found. DNA repeat sequences have been shown to be an interesting potential source of cis-regulatory elements. In this work, a genome-wide search for tandem and palindromic repetitive elements was performed in the intergenic regions of all coding sequences from M. hyopneumoniae strain 7448. Computational analysis demonstrated the presence of 144 tandem repeats and 1,171 palindromic elements. The DNA repeat sequences were distributed within the 5' upstream regions of 86% of transcriptional units of M. hyopneumoniae strain 7448. Comparative analysis between distinct repetitive sequences found in related mycoplasma genomes demonstrated different percentages of conservation among pathogenic and nonpathogenic strains. qPCR assays revealed differential expression among genes showing variable numbers of repetitive elements. In addition, repeats found in 206 genes already described to be differentially regulated under different culture conditions of M. hyopneumoniae strain 232 showed almost 80% conservation in relation to M. hyopneumoniae strain 7448 repeats. Altogether, these findings suggest a potential regulatory role of tandem and palindromic DNA repeats in the M. hyopneumoniae transcriptional profile.

  5. Mycoplasma genitalium: An emerging sexually transmitted pathogen

    PubMed Central

    Sethi, Sunil; Singh, Gagandeep; Samanta, Palash; Sharma, Meera

    2012-01-01

    Mycoplasma genitalium is a member of genital mycoplasmas, which is emerging as an important causative agent of sexually transmitted infections both in males and females. The advent of polymerase chain reaction and other molecular methods have made studies on M. genitalium more feasible, which is otherwise a difficult organism to isolate. Besides Chlamydia trachomatis, M. genitalium is now an important and established cause of non gonococcal urethritis (NGU) in men, more so in persistent and recurrent NGU. Multiple studies have also shown a positive association of M. genitalium with mucopurulent cervicitis and vaginal discharge in females as well. The evidences for M. genitalium pelvic inflammatory diseases and infertility are quite convincing and indicate that this organism has potential to cause ascending infection. Lack of clear association with M. genitalium has been reported for bacterial vaginosis and adverse pregnancy outcomes. Diagnosis of M. genitalium infections is performed exclusively using nucleic acid amplification tests (NAATs), owing to poor or slow growth of bacterium in culture. Although there are no guidelines available regarding treatment, macrolide group of antimicrobials appear to be more effective than tetracyclines. The present review provides an overview of the epidemiology, pathogenesis, clinical presentation and management of sexually transmitted infections due to M. genitalium. PMID:23391789

  6. Repetitive Elements in Mycoplasma hyopneumoniae Transcriptional Regulation

    PubMed Central

    Cattani, Amanda Malvessi; Siqueira, Franciele Maboni; Guedes, Rafael Lucas Muniz; Schrank, Irene Silveira

    2016-01-01

    Transcriptional regulation, a multiple-step process, is still poorly understood in the important pig pathogen Mycoplasma hyopneumoniae. Basic motifs like promoters and terminators have already been described, but no other cis-regulatory elements have been found. DNA repeat sequences have been shown to be an interesting potential source of cis-regulatory elements. In this work, a genome-wide search for tandem and palindromic repetitive elements was performed in the intergenic regions of all coding sequences from M. hyopneumoniae strain 7448. Computational analysis demonstrated the presence of 144 tandem repeats and 1,171 palindromic elements. The DNA repeat sequences were distributed within the 5’ upstream regions of 86% of transcriptional units of M. hyopneumoniae strain 7448. Comparative analysis between distinct repetitive sequences found in related mycoplasma genomes demonstrated different percentages of conservation among pathogenic and nonpathogenic strains. qPCR assays revealed differential expression among genes showing variable numbers of repetitive elements. In addition, repeats found in 206 genes already described to be differentially regulated under different culture conditions of M. hyopneumoniae strain 232 showed almost 80% conservation in relation to M. hyopneumoniae strain 7448 repeats. Altogether, these findings suggest a potential regulatory role of tandem and palindromic DNA repeats in the M. hyopneumoniae transcriptional profile. PMID:28005945

  7. Frequency of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma species in cervical samples.

    PubMed

    Rodrigues, M M; Fernandes, P Á; Haddad, J P; Paiva, M C; Souza, M Do Carmo M; Andrade, T C A; Fernandes, A P

    2011-01-01

    We investigated the relative frequencies of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma sp. in cervical samples. PCR analyses were performed in ectocervical and endocervical samples from 224 patients attending public health services in Belo Horizonte and Contagem, Minas Gerais Brazil. A high prevalence of colonisation of the cervix (6.3% for C. trachomatis, 4.0% for N. gonorrhoeae, 0.9% for M. genitalium, 21.9% for M. hominis, 38.4% for Ureaplasma sp.) was demonstrated not only for pathogens classically associated to cervicitis (C. trachomatis and N. gonorrhoeae), but also for M. hominis and Ureaplasma sp. These findings may be useful to guide more adequate diagnosis to interrupt transmission and to avoid negative impacts on the female reproductive tract.

  8. Characteristics of a new sterol-nonrequiring Mycoplasma.

    PubMed

    Tully, J G; Razin, S

    1969-06-01

    Two Mycoplasma strains recovered from tissue culture environments were found to grow in complex media devoid of serum or serum fractions containing cholesterol and in a cholesterol-free synthetic medium. Neither strain was capable of synthesizing pigmented carotenoids, although these compounds are present in, and characteristic of, other sterol-nonrequiring mycoplasmas. Serological tests and an analysis of their cell protein patterns obtained by gel electrophoresis indicated that the isolates were similar to each other but distinct from other sterol-nonrequiring serotypes, Mycoplasma laidlawii and M. granularum, as well as from sterol-requiring species. The existence of Mycoplasma other than M. laidlawii and M. granularum without sterol requirements suggested the need for some taxonomic changes in this group of organisms.

  9. Genome Sequence of Mycoplasma hyorhinis Strain DBS 1050.

    PubMed

    Dabrazhynetskaya, Alena; Soika, Valerii; Volokhov, Dmitriy; Simonyan, Vahan; Chizhikov, Vladimir

    2014-03-06

    Mycoplasma hyorhinis is known as one of the most prevalent contaminants of mammalian cell and tissue cultures worldwide. Here, we present the complete genome sequence of the fastidious M. hyorhinis strain DBS 1050.

  10. Mycoplasma hominis Induces Mediastinitis after a Tonsillar Abscess

    PubMed Central

    Grancini, Anna; Colosimo, Manuela; Restelli, Antonella; Colombo, Rosaria; Maraschini, Anna; Pozzi, Cristina; Breda, Giuseppe; Protti, Alessandro; Arghittu, Milena; Maiavacca, Rita

    2016-01-01

    Mycoplasma hominis is commonly involved in genitourinary tract infections. We report a 59-year-old man who developed a M. hominis-associated mediastinitis following acute tonsillar infection. PMID:27957362

  11. Mycoplasma pneumoniae induces cytotoxic activity in guinea pig bronchoalveolar cells

    SciTech Connect

    Kist, M.; Koester, H.; Bredt, W.

    1985-06-01

    Precultured guinea pig alveolar macrophages (AM) and freshly harvested alveolar cells (FHAC) activated by interaction with Mycoplasma pneumoniae were cytotoxic for xenogeneic /sup 75/selenomethionine-labeled tumor target cells. Phagocytosis of whole opsonized or nonopsonized M. pneumoniae cells was more effective in eliciting cytotoxicity than uptake of sonicated microorganisms. The addition of living mycoplasma cells to the assay system enhanced the cytotoxic effect considerably. Target cells were significantly more susceptible to the cytotoxic action of phagocytes if they were coated with mycoplasma antigen or cocultured together with M. pneumoniae. The activation of the phagocytes could be inhibited by 2-deoxy-D-glucose but not by antimicrobial substances suppressing mycoplasma protein synthesis. It was accompanied by /sup 51/Cr release without detectable signs of cell damage. The supernatants of activated cells were cytotoxic for approximately 24 h. Inhibition, release, and cytotoxic activity indicate the necessity of an intact metabolism of the effector cells and suggest a secretion of cytotoxic substances.

  12. Urinary tract infections due to Mycoplasma canis in dogs.

    PubMed

    Ulgen, M; Cetin, C; Sentürk, S; Ozel, A E; Ozdemir, U

    2006-09-01

    Urine samples were obtained from 100 dogs with symptoms of lower urinary tract disease by cystocentesis and were examined for mycoplasmas. Urinalysis, haematological and biochemical analyses were also performed. Bacteria were isolated from urine in 41 of 100 dogs; Mycoplasma canis was isolated from four of 100 (4%) urine samples and three were pure culture. Selective mycoplasma media were used for isolation. In growth inhibition test, propagation of the four M. canis isolates was inhibited by their specific hyperimmune sera and there was no cross reactivity between isolates and hyperimmune sera of other mycoplasmas. Dogs in which M. canis was isolated were azotemic. All dogs were treated with enrofloxacin, furosemide, and supportive therapy (fluid therapy, ascorbic acid). In all animals, clinical improvements were observed after treatment.

  13. Susceptibilities of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum strains to antimicrobial agents in vitro.

    PubMed

    ter Laak, E A; Noordergraaf, J H; Verschure, M H

    1993-02-01

    The purpose of this study was to determine the susceptibility of various strains of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum, which are prevalent causes of pneumonia in calves, to 16 antimicrobial agents in vitro. The MICs of the antimicrobial agents were determined by a serial broth dilution method for 16 field strains and the type strain of M. bovis, for 19 field strains and the type strain of M. dispar, and for 17 field strains of U. diversum. Final MICs for M. bovis and M. dispar were read after 7 days and final MICs for U. diversum after 1 to 2 days. All strains tested were susceptible to tylosin, kitasamycin, and tiamulin but were resistant to nifuroquine and streptomycin. Most strains of U. diversum were intermediately susceptible to oxytetracycline but fully susceptible to chlortetracycline; most strains of M. bovis and M. dispar, however, were resistant to both agents. Strains of M. dispar and U. diversum were susceptible to doxycycline and minocycline, but strains of M. bovis were only intermediately susceptible. Susceptibility or resistance to chloramphenicol, spiramycin, spectinomycin, lincomycin, or enrofloxacin depended on the species but was not equal for the three species. The type strains of M. bovis and M. dispar were more susceptible to various antimicrobial agents, including tetracyclines, than the field strains. This finding might indicate that M. bovis and M. dispar strains are becoming resistant to these agents. Antimicrobial agents that are effective in vitro against all three mycoplasma species can be considered for treating mycoplasma infections in pneumonic calves. Therefore, tylosin, kitasamycin, and tiamulin may be preferred over oxytetracycline and chlortetracycline.

  14. Susceptibilities of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum strains to antimicrobial agents in vitro.

    PubMed Central

    ter Laak, E A; Noordergraaf, J H; Verschure, M H

    1993-01-01

    The purpose of this study was to determine the susceptibility of various strains of Mycoplasma bovis, Mycoplasma dispar, and Ureaplasma diversum, which are prevalent causes of pneumonia in calves, to 16 antimicrobial agents in vitro. The MICs of the antimicrobial agents were determined by a serial broth dilution method for 16 field strains and the type strain of M. bovis, for 19 field strains and the type strain of M. dispar, and for 17 field strains of U. diversum. Final MICs for M. bovis and M. dispar were read after 7 days and final MICs for U. diversum after 1 to 2 days. All strains tested were susceptible to tylosin, kitasamycin, and tiamulin but were resistant to nifuroquine and streptomycin. Most strains of U. diversum were intermediately susceptible to oxytetracycline but fully susceptible to chlortetracycline; most strains of M. bovis and M. dispar, however, were resistant to both agents. Strains of M. dispar and U. diversum were susceptible to doxycycline and minocycline, but strains of M. bovis were only intermediately susceptible. Susceptibility or resistance to chloramphenicol, spiramycin, spectinomycin, lincomycin, or enrofloxacin depended on the species but was not equal for the three species. The type strains of M. bovis and M. dispar were more susceptible to various antimicrobial agents, including tetracyclines, than the field strains. This finding might indicate that M. bovis and M. dispar strains are becoming resistant to these agents. Antimicrobial agents that are effective in vitro against all three mycoplasma species can be considered for treating mycoplasma infections in pneumonic calves. Therefore, tylosin, kitasamycin, and tiamulin may be preferred over oxytetracycline and chlortetracycline. PMID:8452363

  15. Functional characterization of the RuvB homologs from Mycoplasma pneumoniae and Mycoplasma genitalium.

    PubMed

    Estevão, Silvia; Sluijter, Marcel; Hartwig, Nico G; van Rossum, Annemarie M C; Vink, Cornelis

    2011-12-01

    Homologous recombination between repeated DNA elements in the genomes of Mycoplasma species has been hypothesized to be a crucial causal factor in sequence variation of antigenic proteins at the bacterial surface. To investigate this notion, studies were initiated to identify and characterize the proteins that form part of the homologous DNA recombination machinery in Mycoplasma pneumoniae as well as Mycoplasma genitalium. Among the most likely participants of this machinery are homologs of the Holliday junction migration motor protein RuvB. In both M. pneumoniae and M. genitalium, genes have been identified that have the capacity to encode RuvB homologs (MPN536 and MG359, respectively). Here, the characteristics of the MPN536- and MG359-encoded proteins (the RuvB proteins from M. pneumoniae strain FH [RuvB(FH)] and M. genitalium [RuvB(Mge)], respectively) are described. Both RuvB(FH) and RuvB(Mge) were found to have ATPase activity and to bind DNA. In addition, both proteins displayed divalent cation- and ATP-dependent DNA helicase activity on partially double-stranded DNA substrates. The helicase activity of RuvB(Mge), however, was significantly lower than that of RuvB(FH). Interestingly, we found RuvB(FH) to be expressed exclusively by subtype 2 strains of M. pneumoniae. In strains belonging to the other major subtype (subtype 1), a version of the protein is expressed (the RuvB protein from M. pneumoniae strain M129 [RuvB(M129)]) that differs from RuvB(FH) in a single amino acid residue (at position 140). In contrast to RuvB(FH), RuvB(M129) displayed only marginal levels of DNA-unwinding activity. These results demonstrate that M. pneumoniae strains (as well as closely related Mycoplasma spp.) can differ significantly in the function of components of their DNA recombination and repair machinery.

  16. Selective inhibition of DNA amplification in nonadhering Mycoplasma pneumoniae cultures

    SciTech Connect

    Zigangirova, N.A.; Solov`eva, S.V.; Rakovskaya, I.V.

    1995-08-01

    Inhibition of amplification of various genome regions of Mycoplasma pneumoniae was observed in the polymerase chain reaction, and was dependent on cultivation conditions. A protein stably associated with DNA is responsible for the inhibitory effect. It is assumed that when the protein selectively associates with separate DNA regions, it can inhibit genes encoding pathogenicity factors, thus promoting mycoplasma transformation into persistent variants. 16 refs., 2 figs.

  17. Mycoplasma insons sp. nov., a twisted mycoplasma from green iguanas (Iguana iguana).

    PubMed

    May, Meghan; Ortiz, G Javier; Wendland, Lori D; Rotstein, David S; Relich, Ryan F; Balish, Mitchell F; Brown, Daniel R

    2007-09-01

    Mycoplasma insons sp. nov., first cultured from the choanae and tracheae of healthy green iguanas (Iguana iguana) from El Salvador, was readily distinguished from all previously described mollicutes and assigned to the Mycoplasma fastidiosum phylogenetic cluster by 16S rRNA gene sequence comparisons. Growth inhibition assays distinguished the isolates serologically from the other two members of that cluster. Many M. insons cells exhibit a remarkable twisted rod morphology despite lacking a cell wall. The organism is nonmotile, produces acid from glucose, but does not hydrolyze arginine, esculin, or urea. Mycoplasma insons 16S rRNA gene was also detected by PCR in packed blood cells from culture-negative iguanas. The type strain I17P1(T) has been deposited with the Mollicutes Collection at Purdue University and with the American Type Culture Collection (ATCC BAA-1435) in the USA. A limited number of cultures generated by the authors have also been deposited with the Culture Collection, University of Göteborg, in Sweden (CCUG 53461).

  18. Complexity of the Mycoplasma fermentans M64 Genome and Metabolic Essentiality and Diversity among Mycoplasmas

    PubMed Central

    Chan, Huang-I; Liu, Yen-Ming; Wu, Keh-Ming; Shu, Hung-Yu; Tsai, Shih-Feng; Hsiao, Kwang-Jen; Hu, Wensi S.; Ng, Wailap Victor

    2012-01-01

    Recently, the genomes of two Mycoplasma fermentans strains, namely M64 and JER, have been completely sequenced. Gross comparison indicated that the genome of M64 is significantly bigger than the other strain and the difference is mainly contributed by the repetitive sequences including seven families of simple and complex transposable elements ranging from 973 to 23,778 bps. Analysis of these repeats resulted in the identification of a new distinct family of Integrative Conjugal Elements of M. fermentans, designated as ICEF-III. Using the concept of “reaction connectivity”, the metabolic capabilities in M. fermentans manifested by the complete and partial connected biomodules were revealed. A comparison of the reported M. pulmonis, M. arthritidis, M. genitalium, B. subtilis, and E. coli essential genes and the genes predicted from the M64 genome indicated that more than 73% of the Mycoplasmas essential genes are preserved in M. fermentans. Further examination of the highly and partly connected reactions by a novel combinatorial phylogenetic tree, metabolic network, and essential gene analysis indicated that some of the pathways (e.g. purine and pyrimidine metabolisms) with partial connected reactions may be important for the conversions of intermediate metabolites. Taken together, in light of systems and network analyses, the diversity among the Mycoplasma species was manifested on the variations of their limited metabolic abilities during evolution. PMID:22509252

  19. Detection of mycoplasma contamination in cell cultures by a mycoplasma group-specific PCR.

    PubMed Central

    van Kuppeveld, F J; Johansson, K E; Galama, J M; Kissing, J; Bölske, G; van der Logt, J T; Melchers, W J

    1994-01-01

    The suitability of a 16S rRNA-based mycoplasma group-specific PCR for the detection of mycoplasma contamination in cell cultures was investigated. A total of 104 cell cultures were tested by using microbiological culture, DNA fluorochrome staining, DNA-rRNA hybridization, and PCR techniques. A comparison of the results obtained with these techniques revealed agreement for 95 cell cultures. Discrepant results, which were interpreted as false negative or false positive on the basis of a comparison with the results obtained with other methods, were observed with nine cell cultures. The microbiological culture technique produced false-negative results for four cell cultures. The hybridization technique produced false-negative results for two cell cultures, and for one of these cell cultures the DNA staining technique also produced a false-negative result. The PCR may have produced false-positive results for one cell culture. Ambiguous results were obtained with the remaining two cell cultures. Furthermore, the presence of contaminating bacteria interfered with the interpretation of the DNA staining results for 16 cell cultures. For the same reason the hybridization signals of nine cell cultures could not be interpreted. Our results demonstrate the drawbacks of each of the detection methods and the suitability of the PCR for the detection of mycoplasmas in cell cultures. PMID:7509584

  20. Prevalence of Mycoplasma genitalium and Mycoplasma hominis in urogenital tract of Brazilian women.

    PubMed

    Campos, Guilherme Barreto; Lobão, Tássia Neves; Selis, Nathan Neves; Amorim, Aline Teixeira; Martins, Hellen Braga; Barbosa, Maysa Santos; Oliveira, Thiago Henrique Caldeira; dos Santos, Djanilson Barbosa; Figueiredo, Tiana Baqueiro; Miranda Marques, Lucas; Timenetsky, Jorge

    2015-02-14

    The role of Mycoplasma hominis and M. genitalium in urogenital tract infections remains unknown. Furthermore these mollicutes present a complex relationship with the host immune response. The role of inflammatory cytokines in infections also makes them good candidates to investigate bacterial vaginosis and mycoplasma genital infections. Therefore, the aim of this study was to detect the above-mentioned mollicutes by quantitative Polymerase Chain Reaction (qPCR) methodologies in vaginal swabs and dosage of cytokines. Vaginal swabs and peripheral blood were collected from 302 women, including healthy individuals. The molecular findings were correlated with some individual behavioral variables, clinical and demographic characteristics, presence of other important microorganisms in vaginal swabs, and levels of interleukin (IL)-1β and IL-6. M. hominis and M. genitalium were detected in 31.8% and 28.1% of samples, respectively. The qPCR results were associated with clinical signs and symptoms of the infections studied. The frequency of Trichomonas vaginalis, Gardnerella vaginalis, Neisseria gonorrhoeae and Chlamydia trachomatis was 3.0%, 21.5%, 42.4%, and 1.7% respectively. Increased levels of IL-1β were associated with the presence of M. hominis and signs and/or symptoms of the genital infection of women studied. IL-1β production was associated with the detection of M. hominis by qPCR. The sexual behavior of women studied was associated with the detection of mycoplasma and other agents of genital infections.

  1. Comparison of the illumigene Mycoplasma DNA Amplification Assay and Culture for Detection of Mycoplasma pneumoniae

    PubMed Central

    Ratliff, Amy E.; Duffy, Lynn B.

    2014-01-01

    A loop-mediated isothermal amplification (LAMP) system, the illumigene Mycoplasma DNA amplification assay (Meridian Bioscience, Inc., Cincinnati, OH) was evaluated to determine its analytical sensitivity, specificity, and clinical application in comparison to historic culture in a collection of archived respiratory specimens. The illumigene limit of detection was ≤88 CFU/reaction for 10 Mycoplasma pneumoniae reference strains. This assay correctly identified 36 M. pneumoniae reference strains and clinical isolates from various geographic origins, including both of the main subtypes. No cross-reactions were detected with other mycoplasmas, ureaplasmas, other bacterial species, viruses, yeasts, or human DNA. Among 214 respiratory specimens previously cultured for M. pneumoniae, when real-time PCR with bidirectional sequencing of the PCR products was used to resolve discrepancies, the sensitivity was 22 of 22 (100%) and the specificity was 190 of 192 (99%). This commercial LAMP assay is a useful rapid method for detecting M. pneumoniae in clinical specimens. Additional prospective clinical trials with direct comparison to culture and PCR are warranted. PMID:24430454

  2. Genomic repeats, genome plasticity and the dynamics of Mycoplasma evolution

    PubMed Central

    Rocha, Eduardo P. C.; Blanchard, Alain

    2002-01-01

    Mycoplasmas evolved by a drastic reduction in genome size, but their genomes contain numerous repeated sequences with important roles in their evolution. We have established a bioinformatic strategy to detect the major recombination hot-spots in the genomes of Mycoplasma pneumoniae, Mycoplasma genitalium, Ureaplasma urealyticum and Mycoplasma pulmonis. This allowed the identification of large numbers of potentially variable regions, as well as a comparison of the relative recombination potentials of different genomic regions. Different trends are perceptible among mycoplasmas, probably due to different functional and structural constraints. The largest potential for illegitimate recombination in M.pulmonis is found at the vsa locus and its comparison in two different strains reveals numerous changes since divergence. On the other hand, the main M.pneumoniae and M.genitalium adhesins rely on large distant repeats and, hence, homologous recombination for variation. However, the relation between the existence of repeats and antigenic variation is not necessarily straightforward, since repeats of P1 adhesin were found to be anti-correlated with epitopes recognized by patient antibodies. These different strategies have important consequences for the structures of genomes, since large distant repeats correlate well with the major chromosomal rearrangements. Probably to avoid such events, mycoplasmas strongly avoid inverse repeats, in comparison to co-oriented repeats. PMID:11972343

  3. A Mycoplasma species of Emydidae turtles in the northeastern USA.

    PubMed

    Ossiboff, Robert J; Raphael, Bonnie L; Ammazzalorso, Alyssa D; Seimon, Tracie A; Niederriter, Holly; Zarate, Brian; Newton, Alisa L; McAloose, Denise

    2015-04-01

    Mycoplasma infections can cause significant morbidity and mortality in captive and wild chelonians. As part of a health assessment of endangered bog turtles (Glyptemys muhlenbergii) in the northeastern US, choanal and cloacal swabs from these and other sympatric species, including spotted turtles (Clemmys guttata), eastern box turtles (Terrapene carolina carolina), wood turtles (Glyptemys insculpta), and common snapping turtles (Chelydra serpentina) from 10 sampling sites in the states (US) of Delaware, New Jersey, and Pennsylvania, were tested by PCR for Mycoplasma. Of 108 turtles tested, 63 (58.3%) were PCR positive for Mycoplasma including 58 of 83 bog turtles (70%), three of three (100%) eastern box turtles, and two of 11 (18%) spotted turtles; all snapping turtles (n = 7) and wood turtles (n = 4) were negative. Sequence analysis of portions of the 16S-23S intergenic spacer region and the 16S ribosomal RNA gene revealed a single, unclassified species of Mycoplasma that has been previously reported in eastern box turtles, ornate box turtles (Terrapene ornata ornata), western pond turtles (Emys marmorata), and red-eared sliders (Trachemys scripta elegans). We document a high incidence of Mycoplasma, in the absence of clinical disease, in wild emydid turtles. These findings, along with wide distribution of the identified Mycoplasma sp. across a broad geographic region, suggest this bacterium is likely a commensal inhabitant of bog turtles, and possibly other species of emydid turtles, in the northeastern US.

  4. Role of Vpma phase variation in Mycoplasma agalactiae pathogenesis

    PubMed Central

    Chopra-Dewasthaly, Rohini; Baumgartner, Martina; Gamper, Erika; Innerebner, Carmen; Zimmermann, Martina; Schilcher, Franz; Tichy, Alexander; Winter, Petra; Rosengarten, Renate; Spergser, Joachim

    2015-01-01

    Compared with other bacterial pathogens, the molecular mechanisms of mycoplasma pathogenicity are largely unknown. Several studies in the past have shown that pathogenic mycoplasmas are equipped with sophisticated genetic systems that allow them to undergo high-frequency surface antigenic variations. Although never clearly proven, these variable mycoplasma surface components are often implicated in host immune evasion and adaptation. Vpma surface lipoproteins of the ruminant pathogen Mycoplasma agalactiae are encoded on a genomic pathogenicity island–like locus and are considered as one of the well-characterized model systems of mycoplasma surface antigenic variation. The present study assesses the role of these phase-variable Vpmas in the molecular pathogenesis of M. agalactiae by testing the wild-type strain PG2 in comparison with the xer1-disrupted Vpma ‘phase-locked’ mutants in sheep infection models. The data clearly illustrate that although Xer1 recombinase is not a virulence factor of M. agalactiae and Vpma phase variation is not necessary for establishing an infection, it might critically influence the survival and persistence of the pathogen under natural field conditions, mainly due to a better capacity for dissemination and evoking systemic responses. This is the first study where mycoplasma ‘phase-locked’ mutants are tested in vivo to elucidate the role of phase variation during infection. PMID:22809092

  5. Mycoplasma Removal from Cell Culture Using Antimicrobial Photodynamic Therapy

    PubMed Central

    Hasebe, Akira; Ishikawa, Isao; Shamsul, Haque M.; Ohtani, Makoto; Segawa, Taku; Saeki, Ayumi; Tanizume, Naoho; Oouchi, Manabu; Okagami, Yoshihide; Okano, Teruo

    2013-01-01

    Abstract Objective: The objective of this research was to determine the effectiveness of antimicrobial photodynamic therapy (aPDT) in the removal of mycoplasmas from contaminated cells. Background data: Mycoplasmas often contaminate cell cultures. The cell-contaminating mycoplasmas are removed by antibiotics, but the use of antibiotics usually induces antibiotic-resistant bacteria. aPDT is expected to be a possible alternative to antibiotic treatments for suppressing infections. Materials and Methods: Mycoplasma salivarium (Ms)-infected human embryonic kidney (HEK) 293 cells were irradiated using a red light-emitting diode (LED) in the presence of methylene blue (MB) as a photosensitizer. The Ms viable count was determined using culture on agar plates or using a mycoplasma detection kit. Results: aPDT performed using red LED irradiation was effective in decreasing live Ms in the presence of MB without damaging the HEK293 cells. aPDT removed live Ms from the infected cells after washing the cells with sterilized phosphate-buffered saline (PBS) to decrease the initial number of live Ms before aPDT. Conclusions: This study suggests that aPDT could remove mycoplasmas from contaminated cells. PMID:23402393

  6. Role of binding in Mycoplasma mobile and Mycoplasma pneumoniae gliding analyzed through inhibition by synthesized sialylated compounds.

    PubMed

    Kasai, Taishi; Nakane, Daisuke; Ishida, Hideharu; Ando, Hiromune; Kiso, Makoto; Miyata, Makoto

    2013-02-01

    Mycoplasmas, which have been shown to be the causative pathogens in recent human pneumonia epidemics, bind to solid surfaces and glide in the direction of the membrane protrusion at a pole. During gliding, the legs of the mycoplasma catch, pull, and release sialylated oligosaccharides fixed on a solid surface. Sialylated oligosaccharides are major structures on animal cell surfaces and are sometimes targeted by pathogens, such as influenza virus. In the present study, we analyzed the inhibitory effects of 16 chemically synthesized sialylated compounds on the gliding and binding of Mycoplasma mobile and Mycoplasma pneumoniae and concluded the following. (i) The recognition of sialylated oligosaccharide by mycoplasma legs proceeds in a "lock-and-key" fashion, with the binding affinity dependent on structural differences among the sialylated compounds examined. (ii) The binding of the leg and the sialylated oligosaccharide is cooperative, with Hill constants ranging from 2 to 3. (iii) Mycoplasma legs may generate a drag force after a stroke, because the gliding speed decreased and pivoting motion occurred more frequently when the number of working legs was reduced by the addition of free sialylated compounds.

  7. Immunological characterization of Mycoplasma hyopneumoniae recombinant proteins.

    PubMed

    Simionatto, Simone; Marchioro, Silvana B; Galli, Vanessa; Brum, Clarice B; Klein, Catia S; Rebelatto, Raquel; Silva, Everton F; Borsuk, Sibele; Conceição, Fabricio R; Dellagostin, Odir A

    2012-03-01

    Mycoplasma hyopneumoniae, the primary pathogen of enzootic pneumonia, is highly prevalent worldwide and causes major economic losses to the pig industry. Commercial vaccines are widely used in the control of this disease, however, they provide only partial protection. The aim of this study was to evaluate 34 recombinant proteins of M. hyopneumoniae expressed in Escherichia coli. Antigenic and immunogenic properties of these proteins were analyzed. For this, the proteins were tested against hyperimmune and convalescent pig sera through ELISA and Western blot. Immunogenicity of the recombinant proteins was evaluated in BALB/c mice following intramuscular inoculation. Most antigens were able to induce a strong immune response and sera from inoculated mice were able to recognize native proteins by cell ELISA and Western blot. Several recombinant proteins were specifically recognized by convalescent pig sera, indicating they are expressed during infection. These data may help to develop more efficacious vaccines against M. hyopneumoniae.

  8. Mycoplasma hyopneumoniae: from disease to vaccine development.

    PubMed

    Simionatto, Simone; Marchioro, Silvana Beutinger; Maes, Dominiek; Dellagostin, Odir Antônio

    2013-08-30

    Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia (EP), a disease that affects swine production worldwide. Vaccination is the most cost-effective strategy for the control and prevention of the disease. Despite efforts to control M. hyopneumoniae infection, significant economic losses in pig production continue to occur. The results of genome-based research have the potential to help understand the biology and pathogenesis of M. hyopneumoniae, and contribute to the development of more effective vaccines and diagnostic tests. In this review, the characteristics of M. hyopneumoniae related to pathogenesis and control measures will be discussed. Special emphasis will be placed on vaccination strategies that have been proposed with the use of reverse vaccinology approaches.

  9. Repetitive DNA sequences in Mycoplasma pneumoniae.

    PubMed Central

    Wenzel, R; Herrmann, R

    1988-01-01

    Two types of different repetitive DNA sequences called RepMP1 and RepMP2 were identified in the genome of Mycoplasma pneumoniae. The number of these repeated elements, their nucleotide sequence and their localization on a physical map of the M. pneumoniae genome were determined. The results show that RepMP1 appears at least 10 times and RepMP2 at least 8 times in the genome. The repeated elements are dispersed on the chromosome and, in three cases, linked to each other by a homologous DNA sequence of 400 bp. The elements themselves are 300 bp (for RepMP1) and 150 bp (for RepMP2) long showing a high degree of homology. One copy of RepMP2 is a translated part of the gene for the major cytadhesin protein P1 which is responsible for the adsorption of M. pneumoniae to its host cell. Images PMID:3138660

  10. Mycoplasma and Chlamydia pneumonia in pediatrics.

    PubMed

    Nelson, Christopher T

    2002-03-01

    Mycoplasma pneumoniae and Chlamydia pneumoniae are common respiratory pathogens in children 5 years of age and older. Although distinctly different in structure, these organisms share similar epidemiologic and clinical characteristics in human infection and disease. Pneumonia caused by these organisms usually occurs after infection of the upper respiratory tract, but may occur in the absence of antecedent upper respiratory infection. Diagnosis of infection with C. pneumoniae and M. pneumoniae is most often based on clinical findings alone, though definitive diagnosis of infection with either organism may be confirmed through serologic methods, culture, and nucleic acid-detection methods such as polymerase chain reaction. Macrolide antibiotics are highly effective in the treatment of infected children, leading to rapid clinical resolution and excellent long-term out-come in the majority of patients.

  11. Cytoskeletal elements in the bacterium Mycoplasma pneumoniae

    NASA Astrophysics Data System (ADS)

    Hegermann, Jan; Herrmann, Richard; Mayer, Frank

    2002-09-01

    Mycoplasma pneumoniae is a pathogenic eubacterium lacking a cell wall. Three decades ago, a "rod", an intracellular cytoskeletal structure, was discovered that was assumed to define and stabilize the elongated cell shape. Later, by treatment with detergent, a "Triton shell" (i.e. a fraction of detergent-insoluble cell material) could be obtained, believed to contain additional cytoskeletal elements. Now, by application of a modified Triton X-100 treatment, we are able to demonstrate that M. pneumoniae possesses a cytoskeleton consisting of a blade-like rod and a peripheral lining located close to the inner face of the cytoplasmic membrane, exhibiting features of a highly regular network. Attached "stalks" may support the cytoplasmic membrane. The rod was connected to the cell periphery by "spokes" and showed a defined ultrastructure. Its proximal end was found to be attached to a wheel-like complex. Fibrils extended from the proximal end of the rod into the cytoplasm.

  12. Comparison of strains of Mycoplasma iowae.

    PubMed

    Rhoades, K R

    1984-01-01

    Comparison of biochemical test results and protein electrophoretic patterns of 21 strains of Mycoplasma iowae indicated that all were similar. Comparison of agglutination test results indicated marked within-species antigenic variation. None of 21 antigens prepared from different strains were effective in demonstrating turkey antibody against five reference strains. Examination of sera from turkeys exposed by intra-air-sac inoculation to two pathogenic strains also indicated antigenic variation. Neither the M. iowae type-strain, Iowa 695, nor the other reference strains were effective in demonstrating antibody against both strains used to expose the turkeys. These findings suggest that antigenic variation may be a major problem in effective serodiagnosis of M. iowae infections.

  13. Biosynthesis of cholesteryl glucoside by Mycoplasma gallinarum.

    PubMed

    Smith, P F

    1971-12-01

    The biosynthesis of cholesteryl glucoside by Mycoplasma gallinarum strain J proceeds by the transfer of glucose from uridine-5'-diphosphoglucose to membrane-bound sterol. Galactose also can be coupled to cholesterol via uridine-5'-diphosphogalactose. The reaction is specific for the uridine-5'-diphospho sugars. Enzymatic activity is associated with the membrane. Treatment of the membrane to remove endogenous sterol inactivates the enzyme. Only sterol which has been bound to the membrane participates in the reaction. The optimum pH is about 8.0, and Mg(2+) is required. The reaction is unaffected by nucleotide triphosphate, uridine-5'-monophosphate, and uridine-5'-diphosphate. Reduction of pH to the optimum for beta-glucosidase in the membrane results in loss of synthesized glucoside. The enzyme is saturated at 0.5 mm uridine-5'-diphosphoglucose. The apparent K(m) of 2.05 x 10(-7) indicates a high affinity of the enzyme for the nucleotide sugar.

  14. [100 years of Mycoplasma--pathogenicity for domestic and farm animals].

    PubMed

    Kirchhoff, H; Runge, M

    1998-10-01

    The paper presents a short description of the history of mycoplasmology, the systematic and characteristics of mycoplasmas and the diseases caused by mycoplasmas in cattle, sheep and goats, swine, poultry, and rats and mice. The pathogenicity of mycoplasmas for cats, dogs and horses is discussed.

  15. Hemotropic mycoplasma in a free-ranging black howler monkey (Alouatta caraya) in Brazil.

    PubMed

    Santos, Leonilda C; Cubilla, Michelle P; de Moraes, Wanderlei; Cubas, Zalmir S; Oliveira, Marcos J; Estrada, Marko; Leutenegger, Christian M; Sykes, Jane E; Lindsay, Leann L; Marcondes, Mary; Barros Filho, Ivan R; Biondo, Alexander W

    2013-07-01

    Hemotropic mycoplasmas are bacteria that infect erythrocytes and cause subclinical infections to life-threatening disease. We describe hemotropic mycoplasma infection in a free-ranging black howler monkey (Alouatta caraya). This is the first molecular detection of a hemotropic mycoplasma in a nonhuman primate from Brazil.

  16. Specific Evolution of F1-Like ATPases in Mycoplasmas

    PubMed Central

    Dautant, Alain; Bouyssou, Guillaume; Labroussaa, Fabien; Sköllermo, Anna; Persson, Anja; Blanchard, Alain; Sirand-Pugnet, Pascal

    2012-01-01

    F1F0 ATPases have been identified in most bacteria, including mycoplasmas which have very small genomes associated with a host-dependent lifestyle. In addition to the typical operon of eight genes encoding genuine F1F0 ATPase (Type 1), we identified related clusters of seven genes in many mycoplasma species. Four of the encoded proteins have predicted structures similar to the α, β, γ and ε subunits of F1 ATPases and could form an F1-like ATPase. The other three proteins display no similarity to any other known proteins. Two of these proteins are probably located in the membrane, as they have three and twelve predicted transmembrane helices. Phylogenomic studies identified two types of F1-like ATPase clusters, Type 2 and Type 3, characterized by a rapid evolution of sequences with the conservation of structural features. Clusters encoding Type 2 and Type 3 ATPases were assumed to originate from the Hominis group of mycoplasmas. We suggest that Type 3 ATPase clusters may spread to other phylogenetic groups by horizontal gene transfer between mycoplasmas in the same host, based on phylogeny and genomic context. Functional analyses in the ruminant pathogen Mycoplasma mycoides subsp. mycoides showed that the Type 3 cluster genes were organized into an operon. Proteomic analyses demonstrated that the seven encoded proteins were produced during growth in axenic media. Mutagenesis and complementation studies demonstrated an association of the Type 3 cluster with a major ATPase activity of membrane fractions. Thus, despite their tendency toward genome reduction, mycoplasmas have evolved and exchanged specific F1-like ATPases with no known equivalent in other bacteria. We propose a model, in which the F1-like structure is associated with a hypothetical X0 sector located in the membrane of mycoplasma cells. PMID:22685606

  17. The RuvA homologues from Mycoplasma genitalium and Mycoplasma pneumoniae exhibit unique functional characteristics.

    PubMed

    Sluijter, Marcel; Estevão, Silvia; Hoogenboezem, Theo; Hartwig, Nico G; van Rossum, Annemarie M C; Vink, Cornelis

    2012-01-01

    The DNA recombination and repair machineries of Mycoplasma genitalium and Mycoplasma pneumoniae differ considerably from those of gram-positive and gram-negative bacteria. Most notably, M. pneumoniae is unable to express a functional RecU Holliday junction (HJ) resolvase. In addition, the RuvB homologues from both M. pneumoniae and M. genitalium only exhibit DNA helicase activity but not HJ branch migration activity in vitro. To identify a putative role of the RuvA homologues of these mycoplasmas in DNA recombination, both proteins (RuvA(Mpn) and RuvA(Mge), respectively) were studied for their ability to bind DNA and to interact with RuvB and RecU. In spite of a high level of sequence conservation between RuvA(Mpn) and RuvA(Mge) (68.8% identity), substantial differences were found between these proteins in their activities. First, RuvA(Mge) was found to preferentially bind to HJs, whereas RuvA(Mpn) displayed similar affinities for both HJs and single-stranded DNA. Second, while RuvA(Mpn) is able to form two distinct complexes with HJs, RuvA(Mge) only produced a single HJ complex. Third, RuvA(Mge) stimulated the DNA helicase and ATPase activities of RuvB(Mge), whereas RuvA(Mpn) did not augment RuvB activity. Finally, while both RuvA(Mge) and RecU(Mge) efficiently bind to HJs, they did not compete with each other for HJ binding, but formed stable complexes with HJs over a wide protein concentration range. This interaction, however, resulted in inhibition of the HJ resolution activity of RecU(Mge).

  18. The RuvA Homologues from Mycoplasma genitalium and Mycoplasma pneumoniae Exhibit Unique Functional Characteristics

    PubMed Central

    Sluijter, Marcel; Estevão, Silvia; Hoogenboezem, Theo; Hartwig, Nico G.; van Rossum, Annemarie M. C.; Vink, Cornelis

    2012-01-01

    The DNA recombination and repair machineries of Mycoplasma genitalium and Mycoplasma pneumoniae differ considerably from those of gram-positive and gram-negative bacteria. Most notably, M. pneumoniae is unable to express a functional RecU Holliday junction (HJ) resolvase. In addition, the RuvB homologues from both M. pneumoniae and M. genitalium only exhibit DNA helicase activity but not HJ branch migration activity in vitro. To identify a putative role of the RuvA homologues of these mycoplasmas in DNA recombination, both proteins (RuvAMpn and RuvAMge, respectively) were studied for their ability to bind DNA and to interact with RuvB and RecU. In spite of a high level of sequence conservation between RuvAMpn and RuvAMge (68.8% identity), substantial differences were found between these proteins in their activities. First, RuvAMge was found to preferentially bind to HJs, whereas RuvAMpn displayed similar affinities for both HJs and single-stranded DNA. Second, while RuvAMpn is able to form two distinct complexes with HJs, RuvAMge only produced a single HJ complex. Third, RuvAMge stimulated the DNA helicase and ATPase activities of RuvBMge, whereas RuvAMpn did not augment RuvB activity. Finally, while both RuvAMge and RecUMge efficiently bind to HJs, they did not compete with each other for HJ binding, but formed stable complexes with HJs over a wide protein concentration range. This interaction, however, resulted in inhibition of the HJ resolution activity of RecUMge. PMID:22666500

  19. Survival and replication of Mycoplasma species in recycled bedding sand and association with mastitis on dairy farms in Utah.

    PubMed

    Justice-Allen, A; Trujillo, J; Corbett, R; Harding, R; Goodell, G; Wilson, D

    2010-01-01

    Mycoplasma spp., usually Mycoplasma bovis, are important bovine pathogens that can cause mastitis, metritis, pneumonia, and arthritis. The currently documented routes of transmission of Mycoplasma spp. are through contaminated milking equipment and by direct animal contact. The existence of environmental sources for Mycoplasma spp. and their role in transmission and clinical disease is poorly characterized. Mycoplasma spp. (confirmed as M. bovis in 2 of 4 samples tested using PCR) was found in recycled bedding sand originating from a dairy experiencing an outbreak of clinical mycoplasma mastitis. Mycoplasma spp. were subsequently found in bedding sand from 2 other dairies whose bulk-tank milk was mycoplasma-positive. The association between the occurrence of Mycoplasma spp. in recycled bedding sand and mycoplasma mastitis in cows was further investigated using a pile of recycled sand from dairy 1. Study objectives included the determination of factors associated with the concentration of Mycoplasma spp. in recycled bedding sand and the duration of survival of mycoplasmas in the sand. We also evaluated the efficacy of 2 disinfectants at 2 different concentrations each for the elimination of Mycoplasma spp. from contaminated sand. Mycoplasma spp. survived in the sand pile for 8 mo. The concentration of Mycoplasma spp. within the sand pile was directly related to temperature and precipitation. It was also positively associated with the growth of gram-negative microorganisms, suggesting the possibility of the formation of a biofilm. Ideal temperatures for replication of Mycoplasma spp. occurred between 15 and 20 degrees C. Moisture in the sand and movement of the sand pile also appeared to play a role in replication of mycoplasmas. We found that 0.5% sodium hypochlorite or 2% chlorhexidine were efficacious in eliminating Mycoplasma spp. from contaminated bedding sand. Recycled bedding sand could be an environmental source of Mycoplasma spp., including M. bovis

  20. The in vitro effect of six antimicrobials against Mycoplasma putrefaciens, Mycoplasma mycoides subsp. mycoides LC and Mycoplasma capricolum subsp. capricolum isolated from sheep and goats in Jordan.

    PubMed

    Al-Momani, W; Nicholas, R A J; Janakat, S; Abu-Basha, E; Ayling, R D

    2006-01-01

    Respiratory disease in sheep and goats is a major problem in Jordan and is often associated with Mycoplasma species. Without effective vaccines, control is mainly by chemotherapy, but the uncontrolled use of antimicrobials has led to concerns about the potential development of antimicrobial resistance. The in vitro effect of chloramphenicol, florfenicol, enrofloxacin, tylosin, erythromycin and oxytetracycline was determined against 32 isolates of Mycoplasma species-M. mycoides subsp. mycoides LC (6), M. capricolum subsp. capricolum (8) and M. putrefaciens (18), all isolated from either nasal swabs or milk, from sheep and goats in different regions of Jordan. The antimicrobial susceptibility showed some Mycoplasma species-specific differences, with M. capricolum subsp. capricolum being more susceptible to tylosin and erythromycin. Chloramphenicol and florfenicol were the least effective for all three Mycoplasma species. No trends or significant differences in antimicrobial susceptibilities were observed between sheep and goat isolates, between milk or nasal swab isolates, or between isolates from different regions of Jordan. Some isolates of M. capricolum subsp. capricolum and M. putrefaciens showed higher MIC levels with oxytetracycline, as did two isolates of M. mycoides subsp. mycoides LC with tylosin, possibly indicating signs of development of antimicrobial resistance.

  1. Mycoplasma iowae: relationships among oxygen, virulence, and protection from oxidative stress.

    PubMed

    Pritchard, Rachel E; Balish, Mitchell F

    2015-03-21

    The poultry-associated bacterium Mycoplasma iowae colonizes multiple sites in embryos, with disease or death resulting. Although M. iowae accumulates in the intestinal tract, it does not cause disease at that site, but rather only in tissues that are exposed to atmospheric O2. The activity of M. iowae catalase, encoded by katE, is capable of rapid removal of damaging H2O2 from solution, and katE confers a substantial reduction in the amount of H2O2 produced by Mycoplasma gallisepticum katE transformants in the presence of glycerol. As catalase-producing bacteria are often beneficial to hosts with inflammatory bowel disease, we explored whether M. iowae was exclusively protective against H2O2-producing bacteria in a Caenorhabditis elegans model, whether its protectiveness changed in response to O2 levels, and whether expression of genes involved in H2O2 metabolism and virulence changed in response to O2 levels. We observed that M. iowae was in fact protective against H2O2-producing Streptococcus pneumoniae, but not HCN-producing Pseudomonas aeruginosa, and that M. iowae cells grown in 1% O2 promoted survival of C. elegans to a greater extent than M. iowae cells grown in atmospheric O2. Transcript levels of an M. iowae gene encoding a homolog of Mycoplasma pneumoniae CARDS toxin were 5-fold lower in cells grown in low O2. These data suggest that reduced O2, representing the intestinal environment, triggers M. iowae to reduce its virulence capabilities, effecting a change from a pathogenic mode to a potentially beneficial one.

  2. Metabolomics reveals mycoplasma contamination interferes with the metabolism of PANC-1 cells.

    PubMed

    Yu, Tao; Wang, Yongtao; Zhang, Huizhen; Johnson, Caroline H; Jiang, Yiming; Li, Xiangjun; Wu, Zeming; Liu, Tian; Krausz, Kristopher W; Yu, Aiming; Gonzalez, Frank J; Huang, Min; Bi, Huichang

    2016-06-01

    Mycoplasma contamination is a common problem in cell culture and can alter cellular functions. Since cell metabolism is either directly or indirectly involved in every aspect of cell function, it is important to detect changes to the cellular metabolome after mycoplasma infection. In this study, liquid chromatography mass spectrometry (LC/MS)-based metabolomics was used to investigate the effect of mycoplasma contamination on the cellular metabolism of human pancreatic carcinoma cells (PANC-1). Multivariate analysis demonstrated that mycoplasma contamination induced significant metabolic changes in PANC-1 cells. Twenty-three metabolites were identified and found to be involved in arginine and purine metabolism and energy supply. This study demonstrates that mycoplasma contamination significantly alters cellular metabolite levels, confirming the compelling need for routine checking of cell cultures for mycoplasma contamination, particularly when used for metabolomics studies. Graphical abstract Metabolomics reveals mycoplasma contamination changes the metabolome of PANC-1 cells.

  3. EPS-I Polysaccharide Protects Mycoplasma pulmonis from Phagocytosis

    PubMed Central

    Shaw, Brandon M.; Daubenspeck, James M.; Simmons, Warren L.; Dybvig, Kevin

    2012-01-01

    Few mycoplasmal polysaccharides have been described and little is known about their role in pathogenesis. The infection of mice with Mycoplasma pulmonis has been utilized in many in vivo and in vitro studies to gain a better understanding of host-pathogen interactions during chronic respiratory infection. Although alveolar macrophages have a primary role in host defense, M. pulmonis is killed inefficiently in vitro. One antiphagocytic factor produced by the mycoplasma is the family of phase- and size-variable Vsa lipoproteins. However, bacteria generally employ multiple strategies for combating host defenses, with capsular polysaccharide often having a key role. We show here that mutants lacking the EPS-I polysaccharide of M. pulmonis exhibit increased susceptibility to binding and subsequent killing by alveolar macrophages. These results give further insight into how mycoplasmas are able to avoid the host immune system and sustain a chronic infection. PMID:23190331

  4. Short Communication - Study on quality and efficacy of commercial tylosin and doxycycline products against local isolates of mycoplasma in broilers.

    PubMed

    Riazuddin, -; Khan, Sarzamin; Imtiaz, Naila; Aslam, Saima; Ahmad, Shakoor; Khan, Hamayun; Rabbani, Masood; Muhammad, Javed; Tanveer, Zafar Iqbal; Ali, Sakhawat

    2017-03-01

    The present study was conducted to investigate the quality and efficacy of commercially available preparations of tylosin and doxycycline available in the local market at Peshawar for poultry. In vitro and in vivo, tests were conducted to check the quality of these antimicrobial drugs. In vitro quality control test was performed by High performance liquid chromatographic (HPLC) and micro dilution method. In vivo, efficacy of the test drugs was checked in broilers infected with Mycoplasma gallisepticum. Results of HPLC indicated that test drug-2 contains doxycycline hydrochloride within specified limits but contain high quantity of active ingredient (Tylosin tartrate 120%). Recovery percentage of test drugs (3, 4, 5) were below the pharmacopoeial limit, which contained low quantity of tylosin tartrate (85%, 87.5%, 85%) respectively however, percent recovery of doxycycline were in the appropriate limits. All the tested drugs were effective against Mycoplasma gallisepticum and showed minimum inhibitory concentration (MIC) at 1.9μg/ml. The in vivo result indicated that all tested drugs decreased morbidity and mortality in infected chicks. The birds treated with test drugs (3 and 5) showed mortality of 9.5%, which was slightly higher than the other test groups. The current study suggested that there are incidences of substandard drugs in Pakistan and the drug regularity authorities should take strict actions against the manufacturing companies.

  5. Occurrence of Urease in T Strains of Mycoplasma

    PubMed Central

    Shepard, Maurice C.; Lunceford, Carl D.

    1967-01-01

    A previously unknown metabolite necessary for growth of T strains of Mycoplasma in artificial culture media has been identified as urea. The source of this metabolite was the mammalian plasma or serum enrichment of the culture medium. Normal horse serum was the most satisfactory native protein enrichment for cultivation of T strains of mycoplasma, and it is believed that its superior performance in agar and fluid culture media is associated with its relatively high urea content (approximately 40 mg/100 ml). T-strain urease activity was maximal at pH 6.0 ± 0.5. This is also the optimal pH for growth of T strains. Substrate concentrations greater than 1.0% urea were inhibitory to growth and urease activity of T-strain organisms, and optimal urea concentrations in fluid media appeared to lie within the range of 0.008 to 0.01 m. This range of urea concentration permitted maximal growth of T-strain organisms without rapid loss of viability due to excessive ammonia accumulation and rise in pH to lethal levels. T strains of Mycoplasma were cultivated in a serum-free fluid medium containing urea as the only added metabolite and nitrogen source. T strains are the only known human mycoplasmas which exhibit urease activity, and this biochemical marker can be employed as an aid in the detection and identification of T strains of Mycoplasma (urease color test) and in distinguishing T strains from other members of the human Mycoplasma group. PMID:6025439

  6. Gliding Motility of Mycoplasma mobile on Uniform Oligosaccharides

    PubMed Central

    Kasai, Taishi; Hamaguchi, Tasuku

    2015-01-01

    ABSTRACT The binding and gliding of Mycoplasma mobile on a plastic plate covered by 53 uniform oligosaccharides were analyzed. Mycoplasmas bound to and glided on only 21 of the fixed sialylated oligosaccharides (SOs), showing that sialic acid is essential as the binding target. The affinities were mostly consistent with our previous results on the inhibitory effects of free SOs and suggested that M. mobile recognizes SOs from the nonreducing end with four continuous sites as follows. (i and ii) A sialic acid at the nonreducing end is tightly recognized by tandemly connected two sites. (iii) The third site is recognized by a loose groove that may be affected by branches. (iv) The fourth site is recognized by a large groove that may be enhanced by branches, especially those with a negative charge. The cells glided on uniform SOs in manners apparently similar to those of the gliding on mixed SOs. The gliding speed was related inversely to the mycoplasma's affinity for SO, suggesting that the detaching step may be one of the speed determinants. The cells glided faster and with smaller fluctuations on the uniform SOs than on the mixtures, suggesting that the drag caused by the variation in SOs influences gliding behaviors. IMPORTANCE Mycoplasma is a group of bacteria generally parasitic to animals and plants. Some Mycoplasma species form a protrusion at a pole, bind to solid surfaces, and glide in the direction of the protrusion. These procedures are essential for parasitism. Usually, mycoplasmas glide on mixed sialylated oligosaccharides (SOs) derived from glycoprotein and glycolipid. Since gliding motility on uniform oligosaccharides has never been observed, this study gives critical information about recognition and interaction between receptors and SOs. PMID:26148712

  7. Mycoplasma genitalium: An Emerging Sexually Transmitted Infection.

    PubMed

    Munoz, Jessian L; Goje, Oluwatosin Jaiyeoba

    2016-01-01

    Mycoplasma genitalium has been recognized as a cause of male urethritis, and there is now evidence suggesting that it causes cervicitis and pelvic inflammatory disease in women. M. genitalium is a slow growing organism, and, with the advent of nucleic acid amplification test (NAAT), more studies are being performed, and knowledge about the pathogenicity of this organism elucidated. With NAAT detection, treatment modalities have been studied, and the next challenge is to determine the most effective antimicrobial therapy. Doxycycline, the first-line antibiotic for urethritis, is largely ineffective in the treatment of M. genitalium and furthermore, resistance to macrolide has also emerged. The most effective drug is Moxifloxacin although there are emerging reports of resistance to it in various parts of the world. This paper not only highlights the current research and knowledge, but also reviews the diversity of health implications on the health of men and women infected with M. genitalium. Alternate antibiotics and the impact of M. genitalium on infertility are areas that require more studies as we continue to research into this microorganism.

  8. Mycoplasma genitalium, an emerging sexually transmitted pathogen.

    PubMed

    Cazanave, C; Manhart, L E; Bébéar, C

    2012-09-01

    Mycoplasma genitalium is a sexually transmitted organism associated with non-gonococcal urethritis in men and several inflammatory reproductive tract syndromes in women such as cervicitis, pelvic inflammatory disease, and infertility. There was evidence for an association of M. genitalium with endometritis and pelvic inflammatory disease (PID), but additional studies are necessary to confirm this. The evidence as to whether M. genitalium can cause adverse pregnancy outcomes such as preterm labor is conflicting. But the authors of some studies on M. genitalium as a cause of infertility have reported this association. This species is very difficult to culture; thus, nucleic acid amplification testing is the only method available for M. genitalium detection. The lack of a cell wall makes M. genitalium intrinsically resistant to antibiotics acting at this level, such as beta-lactams. The treatment of M. genitalium infections is not standardized. Macrolides are recommended, especially single-dose azithromycin; tetracyclines are responsible for a great number of therapeutic failures even no resistance mechanism has yet been demonstrated. Acquired resistance to macrolides and fluoroquinolones leading to therapeutic failure has also been reported. All this raises the issue of the most appropriate therapeutic management and requires drafting diagnostic and therapeutic guidelines for the treatment of M. genitalium infections.

  9. Neonate with Mycoplasma hominis meningoencephalitis given moxifloxacin.

    PubMed

    Wildenbeest, Joanne G; Said, Ines; Jaeger, Bregje; van Hest, Reinier M; van de Beek, Diederik; Pajkrt, Dasja

    2016-11-01

    Mycoplasma hominis is a commensal organism in the genitourinary tract that can cause life-threatening CNS infections in neonates after intrauterine infection or through vertical transmission during birth. We present a case of an 11-day-old neonate presenting with fever and supporting laboratory evidence of a CNS infection. No systemic maternal infection or maternal genitourinary tract infection occurred at the time of delivery. Empirical treatment was initiated, consisting of amoxicillin, cefotaxime, and aciclovir. After clinical deterioration, 16S ribosomal DNA PCR in cerebrospinal fluid detected M hominis, antibiotic treatment was switched to moxifloxacin, and pharmacokinetic data were obtained. This Grand Round illustrates the challenges that exist in the diagnosis and treatment of M hominis meningoencephalitis: bacterial cultures are often negative and recommended empirical antimicrobials do not provide adequate antimicrobial coverage. Optimal antimicrobial treatment regimens for M hominis meningoencephalitis are unknown. Although we describe successful treatment of a neonate with a complicated M hominis meningoencephalitis with moxifloxacin, caution with fluoroquinolone monotherapy (including moxifloxacin) has to be taken into account because resistance to fluoroquinolones has previously been described. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Tissue sequestration of 'Candidatus Mycoplasma turicensis'.

    PubMed

    Novacco, Marilisa; Riond, Barbara; Meli, Marina L; Grest, Paula; Hofmann-Lehmann, Regina

    2013-12-27

    'Candidatus Mycoplasma turicensis' ('Candidatus M. turicensis') is a hemoplasma species that infects felids. It differs from other feline hemoplasma species due to its particular infection kinetics and phylogenetic similarity to rodent hemoplasma species. The lower and shorter bacteremia produced by 'Candidatus M. turicensis' suggests a possible tissue sequestration of the organism. The aim of this study was to explore this possibility. Five specified-pathogen free cats were subcutaneously inoculated with 'Candidatus M. turicensis' and sacrificed 86 days after inoculation. Thirty-one selected organs were collected upon necropsy, and samples were analyzed by real-time Taqman(®) PCR. The humoral immune response was monitored by DnaK ELISA. All five cats had detectable 'Candidatus M. turicensis' loads in the majority (52-100%) of the tested tissues. High 'Candidatus M. turicensis' tissue loads (average 3.46×10(4) copies/10 mg) were detected in the samples. The presence of the organisms in the tissues could not be explained by the blood burdens because the blood of four out of five cats tested PCR-negative at the time of necropsy. This is the first study to describe the distribution of 'Candidatus M. turicensis' in various organs; it also demonstrates that, in contrast to other feline hemoplasma species, significant sequestration of 'Candidatus M. turicensis' occurs in many tissues. These results represent an important step toward the understanding of the pathogenesis of 'Candidatus M. turicensis'.

  11. Selective medium for culture of Mycoplasma hyopneumoniae.

    PubMed

    Cook, Beth S; Beddow, Jessica G; Manso-Silván, Lucía; Maglennon, Gareth A; Rycroft, Andrew N

    2016-11-15

    The fastidious porcine respiratory pathogen Mycoplasma hyopneumoniae has proven difficult to culture since it was first isolated in 1965. A reliable solid medium has been particularly challenging. Moreover, clinical and pathological samples often contain the fast-growing M. hyorhinis which contaminates and overgrows M. hyopneumoniae in primary culture. The aim of this study was to optimise the culture medium for recovery of M. hyopneumoniae and to devise a medium for selection of M. hyopneumoniae from clinical samples also containing M. hyorhinis. The solid medium devised by Niels Friis was improved by use of Purified agar and incorporation of DEAE-dextran. Addition of glucose or neutralization of acidity in liquid medium with NaOH did not improve the final yield of viable organisms or alter the timing of peak viability. Analysis of the relative susceptibility of M. hyopneumoniae and M. hyorhinis strains to four antimicrobials showed that M. hyopneumoniae is less susceptible than M. hyorhinis to kanamycin. This was consistent in all UK and Danish strains tested. A concentration of 2μg/ml of kanamycin selectively inhibited the growth of all M. hyorhinis tested, while M. hyopneumoniae was able to grow. This forms the basis of an effective selective culture medium for M. hyopneumoniae.

  12. Mycoplasma alkalescens demonstrated in bronchoalveolar lavage of cattle in Denmark

    PubMed Central

    Kokotovic, Branko; Friis, Niels F; Ahrens, Peter

    2007-01-01

    Mycoplasma alkalescens is an arginine-metabolizing mycoplasma, which has been found in association with mastitis and arthritis in cattle. Routine bacteriological examination of 17 bronchoalveolar lavage samples from calves with pneumonia in a single herd in Denmark, identified M. alkalescens in eight samples. The organism was found as a sole bacterilogical findings in five of the samples as well as in combination with Mannheimia haemolytica, Haemophilus somni and Salmonella Dublin. This is the first report of isolation of M. alkalescens in Denmark. PMID:17204146

  13. Successive synovial Mycoplasma hominis isolates exhibit apparent antigenic variation.

    PubMed Central

    Olson, L D; Renshaw, C A; Shane, S W; Barile, M F

    1991-01-01

    The expression of surface proteins by 14 successive Mycoplasma hominis isolates obtained from the synovial fluid of a chronically infected septic arthritis patient was examined. Marked differences in the expression of surface proteins, as determined by monoclonal antibodies raised against the first isolate, were observed. However, identical restriction patterns and virtually identical hybridization patterns with probes containing the conserved genes of the Mycoplasma capricolum rRNA operon and the Escherichia coli elongation factor Tu suggest that the protein differences might reflect antigenic variation by M. hominis during infection. Images PMID:1879948

  14. Proposal for 'Candidatus Mycoplasma haemomuris subsp. musculi' in mice, and 'Candidatus Mycoplasma haemomuris subsp. ratti' in rats.

    PubMed

    Harasawa, Ryô; Fujita, Hiromi; Kadosaka, Teruki; Ando, Shuji; Rikihisa, Yasuko

    2015-02-01

    Mycoplasma haemomuris is causative of infectious anaemia or splenomegaly in rodents. We examined the nucleotide sequences of the non-ribosomal genes, rnpB and dnaK, in strains of the species M. haemomuris detected in small field mice and black rats. rnpB nucleotide sequences in strains of the species M. haemomuris isolated from small field mice and black rats had only 89 % sequence similarity, suggesting their separation into two distinct subgroups. dnaK had a nucleotide sequence similarity of 84 % between the subgroups. These results support the classification of M. haemomuris into two genetically distinct subgroups. Here we propose the establishment of these subgroups as 'Candidatus Mycoplasma haemomuris subsp. musculi', detected in small field mice (Apodemus argenteus), and 'Candidatus Mycoplasma haemomuris subsp. ratti', detected in black rats (Rattus rattus).

  15. Experimental pathology of T-2 toxicosis and mycoplasma infection on performance and hepatic functions of broiler chickens.

    PubMed

    Manafi, M; Pirany, N; Noor Ali, M; Hedayati, M; Khalaji, S; Yari, M

    2015-07-01

    This experiment was conducted using 192 day-old Ross 308 chicks, divided into 4 groups of 4 replicate consisting 48 birds. Group I was fed a control diet, Group II was fed control diet supplemented with 0.5 ppm T-2 toxin for 5 weeks, Group III was fed control diet supplemented with 8 × 10(8) cfu/mL of Mycoplasma gallisepticum, and group IV was fed control diet supplemented by T-2 toxin and Mycoplasma gallisepticum. Body weight and feed conversation ratio (FCR), relative organ weights, clinical signs, biochemical characteristics, and gross and histopathological lesions were recorded in the experimental groups at the end of the second and fifth weeks of age. Body weight and relative weights of bursa of Fabricius, thymus, and spleen decreased and FCR increased significantly (P ≤ 0.05), but the relative weights of liver and kidney showed no significant decrease (P ≤ 0.05) in the serum total proteins, albumin, and increase in aspartate aminotransferase and alanine transaminase were observed in T-2 toxin and T-2 accompanied with Mycoplasma fed birds when compared to the control group. Liver was enlarged, friable, and yellowish discoloration with distended gall bladder was noticed. Lymphoid organs such as bursa of Fabricius, thymus, and spleen were atrophied in group II and group IV throughout the study. Microscopically, liver showed vacuolar degeneration of hepatocytes, with increased Kupffer cell activity, bile duct epithelial hyperplasia, and infiltration of inflammatory cells. Kidney showed vacuolar degeneration of tubular epithelium along with pyknotic nuclei. Lymphoid organs showed lymphocytolysis and depletion with prominent reticuloepithelial cells. Proventriculus revealed desquamation of villous epithelial cells and lymphoid infiltration in submucosa. Heart showed mild hemorrhage with infiltration of inflammatory cells. Lung showed edema and inflammatory cells in the bronchioles. Trachea showed desquamation and erosions of mucosa. Proliferation of mucosal

  16. Experimental studies on the pathogenicity of Mycoplasma ovipneumoniae and Mycoplasma arginini for the respiratory tract of goats.

    PubMed Central

    Goltz, J P; Rosendal, S; McCraw, B M; Ruhnke, H L

    1986-01-01

    Mycoplasma ovipneumoniae and Mycoplasma arginini were the species of Mollicutes most commonly isolated from 175 goats with respiratory disease in Ontario. The pathogenicity of M. ovipneumoniae, strain B321B and M. arginini, strain D53e, was assessed in goats following endobronchial inoculation. One out of three two year old goats developed fever after inoculation with a pure culture of strain B321B, and it had extensive subacute fibrinous pleuritis when necropsied three weeks later. Neither of the remaining goats had lesions in the respiratory tract. Mycoplasma ovipneumoniae was recovered from one of the animals four days after inoculation, but not at necropsy from any of the goats, at which time a marked humoral immune response with growth inhibiting antibodies was detected. In a second experiment three four to five week old goats were inoculated with the same strain and three other goats were given placebo treatment. One experimental goat developed fever and coughing, and it had extensive subacute fibrinous pleuritis in the right side and pneumonia. Another goat had focal pneumonia in the left diaphragmatic lobe. Microscopically there was subacute hyperplastic suppurative bronchiolitis, atelectasis and nonsuppurative alveolitis. The infected animals did not clear the mycoplasma and not all of them produced antibodies. Mycoplasma arginini, strain D53e, did not induce lesions in any of four goat kids within 14 days after inoculation but did cause transient elevations in rectal temperature, circulating monocytes, circulating neutrophils and blood fibrinogen. Mycoplasma arginini was infective and immunogenic for all inoculated animals and showed a particular affinity for the tonsil. Thus, this study provides the first evidence that M. ovipneumoniae is pathogenic for goats causing pneumonia and pleuritis.(ABSTRACT TRUNCATED AT 250 WORDS) Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:3742358

  17. A novel rapid DNA microarray assay enables identification of 37 Mycoplasma species and highlights multiple Mycoplasma infections.

    PubMed

    Schnee, Christiane; Schulsse, Samuel; Hotzel, Helmut; Ayling, Roger D; Nicholas, Robin A J; Schubert, Evelyn; Heller, Martin; Ehricht, Ralf; Sachse, Konrad

    2012-01-01

    Mycoplasmas comprise a conglomerate of pathogens and commensals occurring in humans and animals. The genus Mycoplasma alone contains more than 120 species at present, and new members are continuously being discovered. Therefore, it seems promising to use a single highly parallel detection assay rather than develop separate tests for each individual species. In this study, we have designed a DNA microarray carrying 70 oligonucleotide probes derived from the 23S rRNA gene and 86 probes from the tuf gene target regions. Following a PCR amplification and biotinylation step, hybridization on the array was shown to specifically identify 31 Mycoplasma spp., as well as 3 Acholeplasma spp. and 3 Ureaplasma spp. Members of the Mycoplasma mycoides cluster can be recognized at subgroup level. This procedure enables parallel detection of Mollicutes spp. occurring in humans, animals or cell culture, from mono- and multiple infections, in a single run. The main advantages of the microarray assay include ease of operation, rapidity, high information content, and affordability. The new test's analytical sensitivity is equivalent to that of real-time PCR and allows examination of field samples without the need for culture. When 60 field samples from ruminants and birds previously analyzed by denaturing-gradient gel electrophoresis (DGGE) were tested by the microarray assay both tests identified the same agent in 98.3% of the cases. Notably, microarray testing revealed an unexpectedly high proportion (35%) of multiple mycoplasma infections, i.e., substantially more than DGGE (15%). Two of the samples were found to contain four different Mycoplasma spp. This phenomenon deserves more attention, particularly its implications for epidemiology and treatment.

  18. Anatomic location of Mycoplasma mycoides subsp. capri and Mycoplasma agalactiae in naturally infected goat male auricular carriers.

    PubMed

    Gómez-Martín, Angel; De la Fe, Christian; Amores, Joaquín; Sánchez, Antonio; Contreras, Antonio; Paterna, Ana; Buendía, Antonio J; Corrales, Juan C

    2012-06-15

    This study sought to determine whether male goat auricular carriers of mycoplasmas known to cause contagious agalactia could harbour these microorganisms at anatomical sites other than the ears. A microbiological study was conducted in 6 naturally infected bucks that had been diagnosed as chronic auricular asymptomatic carriers of Mycoplasma (M.) mycoides subsp. capri (Mmc) more than one year previously. To detect mycoplasmas, cultures and PCR were performed on 46 samples taken from each goat from the cardio-respiratory, digestive, nervous, lymph and genitourinary systems and several joints. Of a total of 274 samples analyzed, 28 were positive for mycoplasmas (10.1%): Mmc was detected in 17 (6.1%), Mycoplasma (M.) agalactiae in 12 (4.3%) and both microorganisms were identified in one of the samples. In all 6 goats, mixed infection was observed despite none being auricular carriers of M. agalactiae. Mycoplasma spp. were identified at 15 different sites; the most frequent sites being the joints (31.2%, 5 positive samples), lymph nodes (25%, 4 positive samples) and respiratory tract (25%, 4 positive samples). Positive results were also obtained in three brain tissue (18.7%), two cardiac tissue (12.5%) and one ileum, urethra, testicle and bulbourethral gland (6.25%) samples. The histopathological findings may suggest the presence of mild chronic conditions in some of the organs where the bacteria were found. Our findings reveal for the first time the capacity of Mmc and M. agalactiae to colonize several other organ systems in chronically naturally infected auricular carriers, possibly representing an added risk factor for the spread of these microorganisms. In the case of M. agalactiae, colonization seemed to be independent of the animal's auricular carrier state.

  19. Nanotransformation of the haemotrophic Mycoplasma suis during in vitro cultivation attempts using modified cell free Mycoplasma media.

    PubMed

    Schreiner, Sabrina A; Hoelzle, Katharina; Hofmann-Lehmann, Regina; Hamburger, Anja; Wittenbrink, Max M; Kramer, Manuela M; Sokoli, Albina; Felder, Kathrin M; Groebel, Katrin; Hoelzle, Ludwig E

    2012-11-09

    Mycoplasma suis belongs to haemotrophic mycoplasmas (HMs) which cause infectious anaemia in a large variety of mammals. To date, no in vitro cultivation system for M. suis or other HMs has been established. We hypothesised that M. suis could grow in classical Mycoplasma media supplemented with nutrients (e.g. glucose, iron-binding proteins) which are naturally available from its host environment, the porcine blood. Blood from experimentally M. suis-infected pigs was used to inoculate either standard SP-4 Mycoplasma medium supplemented with iron-binding proteins (transferrin, haemin, and haemoglobin) or glucose-enriched Hayflick Mycoplasma medium. A quantitative M. suis-specific real-time PCR assay was applied to determine and quantify M. suis loads weekly during 12 week-incubation. The first 2 weeks after inoculation M. suis loads decreased remarkably and then persisted at a stationary level over the observation time of 12 weeks in iron-binding protein- or glucose supplemented media variants. Scanning electron microscopic analysis of liquid M. suis sub-cultures on Hayflick agar showed small, densely-packed microcolonies of irregular M. suis cells of reduced size (0.2-0.6μm) indicating nanotransformation. The partial 16S rDNA sequence of these cultured M. suis nanocells was 99.9% identical to M. suis. M. suis cells derived from liquid cultures interact in vitro with porcine erythrocytes by fibril-like structures. We conclude, that the modified Mycoplasma media used for M. suis cultivation are obviously unfavourable for growth but lead to culture persistence. M. suis adapt to inappropriate culture conditions by alteration into nanoforms.

  20. A Novel Rapid DNA Microarray Assay Enables Identification of 37 Mycoplasma Species and Highlights Multiple Mycoplasma Infections

    PubMed Central

    Schnee, Christiane; Schulsse, Samuel; Hotzel, Helmut; Ayling, Roger D.; Nicholas, Robin A. J.; Schubert, Evelyn; Heller, Martin; Ehricht, Ralf; Sachse, Konrad

    2012-01-01

    Mycoplasmas comprise a conglomerate of pathogens and commensals occurring in humans and animals. The genus Mycoplasma alone contains more than 120 species at present, and new members are continuously being discovered. Therefore, it seems promising to use a single highly parallel detection assay rather than develop separate tests for each individual species. In this study, we have designed a DNA microarray carrying 70 oligonucleotide probes derived from the 23S rRNA gene and 86 probes from the tuf gene target regions. Following a PCR amplification and biotinylation step, hybridization on the array was shown to specifically identify 31 Mycoplasma spp., as well as 3 Acholeplasma spp. and 3 Ureaplasma spp. Members of the Mycoplasma mycoides cluster can be recognized at subgroup level. This procedure enables parallel detection of Mollicutes spp. occurring in humans, animals or cell culture, from mono- and multiple infections, in a single run. The main advantages of the microarray assay include ease of operation, rapidity, high information content, and affordability. The new test's analytical sensitivity is equivalent to that of real-time PCR and allows examination of field samples without the need for culture. When 60 field samples from ruminants and birds previously analyzed by denaturing-gradient gel electrophoresis (DGGE) were tested by the microarray assay both tests identified the same agent in 98.3% of the cases. Notably, microarray testing revealed an unexpectedly high proportion (35%) of multiple mycoplasma infections, i.e., substantially more than DGGE (15%). Two of the samples were found to contain four different Mycoplasma spp. This phenomenon deserves more attention, particularly its implications for epidemiology and treatment. PMID:22479374

  1. Identification of Mycoplasma bovigenitalium and Mycoplasma canadense from outbreaks of granulopapular vulvovaginitis in dairy cattle in Israel.

    PubMed

    Lysnyansky, I; Brenner, J; Alpert, N; Benjamin, A; Bernstein, M; Elad, D; Blum, S; Friedgut, O; Rotenberg, D

    2009-09-12

    A syndrome in which white foci and granulopustular lesions appeared on the vaginal mucous membranes of Holstein cows in several dairy herds in Israel is described. During clinical and diagnostic investigations, Mycoplasma bovigenitalium was isolated from 11 of 20 clinical cases. Vaginal swabs taken from the same cows yielded three isolates of Mycoplasma canadense, which were all associated with the M bovigenitalium infection. Two isolates of small, round, non-enveloped viral particles were approximately 25 nm in diameter and characteristic of enteroviruses on negative-staining electron microscopy.

  2. [Identification of species of Mycoplasma and Ureaplasma diversum from Argentinian dairy herds].

    PubMed

    Sosa, Camila; Tirante, Liliana; Chaves, Javier; Pol, Martín; Ambrogi, Arnaldo; Giraudo, José Angel; Tamiozzo, Pablo

    2017-09-27

    Several species of Mycoplasma and Ureaplasma diversum can cause diseases in dairy cattle, which can be associated or not with clinical manifestations. In our country, the presence of Mycoplasma bovis, Mycoplasma californicum and Mycoplasma canadense has been detected, being the only mycoplasma species identified so far. The objective of this study was to identify other species of the Mycoplasmataceae family. Thirty-five Mycoplasma spp.-like isolates obtained from different samples from cattle, with or without clinical symptoms, from eight herds located in the provinces of Santa Fe, Cordoba, Buenos Aires and San Luis were utilized in the present study. Through the use of species-specific polymerase chain reactions (PCR) Mycoplasma bovigenitalium, Mycoplasma alkalescens, Mycoplasma bovirhinis and U. diversum were identified and through amplification and further sequencing of the 16-23S rRNA intergenic spacer regions, Mycoplasma arginine and M. californicum were identified. The identification of these species represents an important advance in knowledge in order to include these pathogens in the differential diagnosis of certain clinical and pathological entities of cattle from Argentina. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  3. Control of Mycoplasma hyopneumoniae infections in pigs.

    PubMed

    Maes, D; Segales, J; Meyns, T; Sibila, M; Pieters, M; Haesebrouck, F

    2008-01-25

    Mycoplasma hyopneumoniae, the primary pathogen of enzootic pneumonia, occurs worldwide and causes major economic losses to the pig industry. The organism adheres to and damages the ciliated epithelium of the respiratory tract. Affected pigs show chronic coughing, are more susceptible to other respiratory infections and have a reduced performance. Control of the disease can be accomplished in a number of ways. First, management practices and housing conditions in the herd should be optimized. These include all-in/all-out production, limiting factors that may destabilize herd immunity, maintaining optimal stocking densities, prevention of other respiratory diseases, and optimal housing and climatic conditions. Strategic medication with antimicrobials active against M. hyopneumoniae and, preferably, also against major secondary bacteria may be useful during periods when the pigs are at risk for respiratory disease. Finally, commercial bacterins are widely used to control M. hyopneumoniae infections. The main effects of vaccination include less clinical symptoms, lung lesions and medication use, and improved performance. However, bacterins provide only partial protection and do not prevent colonization of the organism. Different vaccination strategies (timing of vaccination, vaccination of sows, vaccination combined with antimicrobial medication) can be used, depending on the type of herd, the production system and management practices, the infection pattern and the preferences of the pig producer. Research on new vaccines is actively occurring, including aerosol and feed-based vaccines as well as subunit and DNA vaccines. Eradication of the infection at herd level based on age-segregation and medication is possible, but there is a permanent risk for re-infections.

  4. 21 CFR 864.2360 - Mycoplasma detection media and components.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Mycoplasma detection media and components. 864.2360 Section 864.2360 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Cell And Tissue Culture...

  5. In Vitro Susceptibilities of Mycoplasma hyopneumoniae Field Isolates

    PubMed Central

    Vicca, J.; Stakenborg, T.; Maes, D.; Butaye, P.; Peeters, J.; de Kruif, A.; Haesebrouck, F.

    2004-01-01

    The in vitro susceptibilities of 21 Mycoplasma hyopneumoniae field isolates were determined using a broth microdilution technique. One isolate showed acquired resistance to lincomycin, tilmicosin, and tylosin, while five isolates were resistant to flumequine and enrofloxacin. Acquired resistance against these antimicrobials in M. hyopneumoniae field isolates was not reported previously. PMID:15504886

  6. Atypical pneumonia associated with a Mycoplasma isolate in a kitten.

    PubMed

    Bongrand, Yannick; Blais, Marie-Claude; Alexander, Kate

    2012-10-01

    An atypical case of Mycoplasma pneumonia with an unusual radiographic and computed tomographic pattern was diagnosed in a Siamese kitten. The cat showed no response to broad-spectrum antibiotic therapy including enrofloxacin. The administration of doxycycline led to a dramatic clinical and radiographic improvement.

  7. Interaction of Mycoplasma dispar with bovine alveolar macrophages.

    PubMed Central

    Almeida, R A; Wannemuehler, M J; Rosenbusch, R F

    1992-01-01

    The capacity to avoid phagocytosis and the activation of bovine alveolar macrophages (BAM) by encapsulated Mycoplasma dispar or purified M. dispar capsule was investigated. Encapsulated and unencapsulated M. dispar were cocultured with BAM in the presence or absence of antisera prepared against unencapsulated M. dispar or purified capsule antiserum. Unopsonized mycoplasmas resisted phagocytosis, while only anti-capsule antibodies enhanced the phagocytosis of encapsulated mycoplasmas. BAM were cultured in the presence of purified M. dispar capsule or either live or heat-killed encapsulated or unencapsulated M. dispar. These BAM were then activated with Escherichia coli endotoxin or left without further activation. The supernatants of these cultures were assayed for tumor necrosis factor, interleukin 1, and glucose consumption as indicators of macrophage activation. Tumor necrosis factor and interleukin 1 were produced by BAM stimulated with unencapsulated M. dispar but not when encapsulated M. dispar or its purified capsule was used. Similarly, glucose consumption was increased in the presence of unencapsulated M. dispar, but not when BAM were cocultured with encapsulated M. dispar or purified capsule. When BAM were treated with purified capsule or encapsulated mycoplasmas, they could not be subsequently activated by endotoxin. These results indicate that encapsulated M. dispar or purified capsule exerts an inhibitory effect on the activity of BAM and prevents the activation of these cells. PMID:1612758

  8. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Test for Mycoplasma. 610.30 Section 610.30 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS... Methylene Blue-Azure dye or an equivalent staining procedure is used, no less than a one square cm. plug...

  9. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 7 2013-04-01 2013-04-01 false Test for Mycoplasma. 610.30 Section 610.30 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS... Methylene Blue-Azure dye or an equivalent staining procedure is used, no less than a one square cm. plug...

  10. High Prevalence of Mycoplasma faucium DNA in the Human Oropharynx.

    PubMed

    Edouard, Sophie; Courtois, Gaëlle Denis; Gautret, Philippe; Jouve, Jean-Luc; Minodier, Philippe; Noël, Guilhem; Roch, Antoine; Brouqui, Philippe; Stein, Andreas; Drancourt, Michel; Fournier, Pierre-Edouard; Raoult, Didier

    2016-01-01

    Mycoplasma faucium has recently been associated with brain abscesses and seems to originate from the mouth. We evaluated its prevalence by quantitative real-time PCR (qPCR) in the oropharynxes of 644 subjects and found that 25% harbored M. faucium, probably constituting the gateway for entrance of the bacteria into cerebral abscesses.

  11. Methyl-prednisolone in neurologic complications of Mycoplasma pneumonia.

    PubMed

    Gücüyener, K; Simşek, F; Yilmaz, O; Serdaroğlu, A

    2000-06-01

    In patients with Mycoplasma pneumonia extrapulmonary manifestations such as encephalitis, meningitis, cerebellar and brain stem involvement, cranial nerve lesions, peripheral neuropathy, polymyositis have been observed. We report a 16-year-old girl with M. pneumonia infection, acute behavioral changes and coma. Treatment with high dose methyl-prednisolone and clarithromycin led to rapid clinical improvement.

  12. Cranial neuropathy, myeloradiculopathy, and myositis: complications of Mycoplasma pneumoniae infection.

    PubMed

    Rothstein, T L; Kenny, G E

    1979-08-01

    Polymyositis, transverse myelitis, ascending polyneuritis, bilateral optic neuritis, and hearing loss developed in a patient with high complement-fixing antibody titers to Mycoplasma pneumoniae. Each of her three children had primary atypical pneumonia with isolation of the organism. The neurologic disturbance is thought to represent a postinfectious complication of M pneumoniae infection.

  13. Isolation of Mycoplasma hyosynoviae from pneumonic lung of swine.

    PubMed

    Dahlia, H; Tan, L J; Zarrahimah, Z; Maria, J

    2009-12-01

    The isolation of Mycoplasma hyosynoviae from a piglet with severe pneumonia is described. This is the first report of M. hyosynoviae isolation in the country. The lung sample where the isolation was made was severely consolidated, suppurative and pleurisy. The pathogenicity of the M. hyosynoviae isolated has yet to be determined.

  14. High Prevalence of Mycoplasma faucium DNA in the Human Oropharynx

    PubMed Central

    Edouard, Sophie; Courtois, Gaëlle Denis; Gautret, Philippe; Jouve, Jean-Luc; Minodier, Philippe; Noël, Guilhem; Roch, Antoine; Brouqui, Philippe; Stein, Andreas; Drancourt, Michel; Fournier, Pierre-Edouard

    2015-01-01

    Mycoplasma faucium has recently been associated with brain abscesses and seems to originate from the mouth. We evaluated its prevalence by quantitative real-time PCR (qPCR) in the oropharynxes of 644 subjects and found that 25% harbored M. faucium, probably constituting the gateway for entrance of the bacteria into cerebral abscesses. PMID:26511735

  15. Ureaplasma urealyticum, Mycoplasma hominis and adverse pregnancy outcomes.

    PubMed

    Capoccia, Romina; Greub, Gilbert; Baud, David

    2013-06-01

    Mycoplasma hominis and Ureaplasma urealyticum may colonize the human genital tract and have been associated with adverse pregnancy outcomes. Chorioamnionitis, spontaneous preterm labour and preterm premature rupture of membranes are significant contributors to neonatal morbidity and mortality. However, as these bacteria can reside in the normal vaginal flora, there are controversies regarding their true role during pregnancy and thus the need to treat these organisms. We review here the recent data on the epidemiology of mycoplasmas and their clinical role during pregnancy. The association of these organisms with preterm labour has been suggested by many observational studies, but proof of causality remains limited. PCR is an excellent alternative to culture to detect the presence of these organisms, but culture allows antibiotic susceptibility testing. Whether antimicrobial treatment of mycoplasma-colonized pregnant patients can effectively reduce the incidence of adverse pregnancy outcomes warrants further investigations. The role of Mycoplasma spp. and U. urealyticum in adverse pregnancy outcomes is increasingly accepted. However, sole presence of these microorganisms in the vaginal flora might be insufficient to cause pathological issues, but their combination with other factors such as bacterial vaginosis or cervical incompetence may be additionally needed to induce preterm birth.

  16. Rhabdomyolysis associated with antimicrobial drug-resistant Mycoplasma pneumoniae.

    PubMed

    Oishi, Tomohiro; Narita, Mitsuo; Ohya, Hitomi; Yamanaka, Takayuki; Aizawa, Yuta; Matsuo, Mai; Matsunaga, Masamichi; Tsukano, Shinya; Taguchi, Testuo

    2012-05-01

    We describe a case of rhabdomyolysis in a patient infected with antimicrobial drug-resistant Mycoplasma pneumoniae The patient's acute-phase serum levels of interleukin-18 and tumor necrosis factor-α were high, which suggests a pathogenic role for M. pneumoniae. In an era of increasing antimicrobial drug resistance, a system for rapidly identifying resistant M. pneumoniae would be beneficial.

  17. Mycoplasma mastitis in cattle: To cull or not to cull.

    PubMed

    Nicholas, Robin A J; Fox, Larry K; Lysnyansky, Inna

    2016-10-01

    Bovine mastitis caused by mycoplasmas, in particular Mycoplasma bovis, is a major problem for milk production and animal welfare in large dairy herds in the USA and a serious, although sporadic, disease in Europe and the Middle East. It causes severe damage to the udder of cattle and is largely untreatable by chemotherapy. Mycoplasma mastitis has a distinct epidemiology and a unique set of risk factors, the most important of which is large herd size. The disease is often self-limiting, disappearing within months of outbreaks, sometimes without deliberate intervention. Improved molecular diagnostic tests are leading to more rapid detection of mycoplasmas. Typing tests, such as multi-locus sequence typing, can help trace the source of outbreaks. An approach to successful control is proposed, which involves regular monitoring and rapid segregation or culling of infected cows. Serious consideration should be given by owners of healthy dairy herds to the purchase of M. bovis-free replacements. Increased cases of disease could occur in Europe and Israel if the trend for larger dairy herds continues. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Macrolide susceptibility of Mycoplasma hyorhinis isolated from piglets.

    PubMed Central

    Kobayashi, H; Morozumi, T; Munthali, G; Mitani, K; Ito, N; Yamamoto, K

    1996-01-01

    Twenty strains of Mycoplasma hyorhinis were investigated for their in vitro susceptibilities to 15 antimicrobial agents by broth and agar dilution methods. Two of the 20 field strains showed low susceptibility to 14- and 16-membered macrolide antimicrobial agents tested. The two field strains were considered inducibly resistant to macrolides. PMID:8849222

  19. Mycoplasma bovis: an emerging pathogen of ranched bison

    USDA-ARS?s Scientific Manuscript database

    Mycoplasma bovis (M. bovis) is an emerging, primary pathogen of ranched bison (Bison bison) in North America. It causes severe disease among animals in feedlots as well as breeding-age cows and bulls on pasture. Mortality in adult bison is as high as 25 percent, resulting in significant economic l...

  20. 9 CFR 113.408 - Avian mycoplasma antigen.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Avian mycoplasma antigen. 113.408 Section 113.408 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF... with a dye acceptable to Animal and Plant Health Inspection Service (APHIS). Final container samples of...

  1. 9 CFR 113.408 - Avian mycoplasma antigen.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Avian mycoplasma antigen. 113.408 Section 113.408 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF... with a dye acceptable to Animal and Plant Health Inspection Service (APHIS). Final container samples of...

  2. Mycoplasmas isolated from the respiratory tract of horses.

    PubMed Central

    Allam, N. M.; Lemcke, R. M.

    1975-01-01

    Ten mycoplasmas were isolated from 130 nasopharyngeal swabs from thoroughbred horses with acute respiratory disease and three from 198 apparently normal horses. Two mycoplasmas were isolated from 21 tracheal swabs taken at necropsy. These mycoplasmas, together with six isolated from the equine respiratory tract by other workers, were subjected to biochemical and serological tests. Other properties examined in certain representative strains were appearance under the electron microscope, ability to adsorb or agglutinate the erythrocytes of various animal species and the electrophoretic pattern of the cell proteins. On the basis of these test, mycoplasmas from the equine respiratory tract were divided into seven species. Three species belonged to the genus Acholeplasma, members of which do not require sterol for growth, and were identified as A. laidlawii, A. oculi (formerly A. oculusi) originally isolated from the eyes of goats, and a recently named species A. equifoetale, previously isolated from aborted equine fetuses. Of the four sterol-dependent Mycoplasma species, one was indentified as M. pulmonis, a common rodent pathogen. Another cross-reacted serologically with M. felis and should probably be classified as that species. The other two species probably represent new species peculiar to the horse. One of these, represented by the strains N3 and N11, ferments glucose and is serologically distinct from 19 recognized species of glucose-utilizing mycoplasmas and from two species which do not metabolize either glucose or arginine. The other species, represented by four strains, hydrolyses arginine and, because it is serologically distinct from all the named arginine-hydrolysing Mycoplasma species, the name M. equirhinis sp.nov. is proposed for it. Of the seven species, only M. pulmonis and the glucose-utilizing species represented by N3 and N11 were found exclusively in horses with acute respiratory disease. A. oculi was isolated from an apparently normal horse. The

  3. Biosecurity and geospatial analysis of mycoplasma infections in poultry farms at Al-Jabal Al-Gharbi region of Libya

    PubMed Central

    Kammon, Abdulwahab; Mulatti, Paolo; Lorenzetto, Monica; Ferre, Nicola; Sharif, Monier; Eldaghayes, Ibrahim; Dayhum, Abdunaser

    2017-01-01

    Geospatial database of farm locations and biosecurity measures are essential to control disease outbreaks. A study was conducted to establish geospatial database on poultry farms in Al-Jabal Al-Gharbi region of Libya, to evaluate the biosecurity level of each farm and to determine the seroprevalence of mycoplasma and its relation to biosecurity level. A field team of 7 Veterinarians belongs to the National Center of Animal Health was assigned for data recording and collection of blood samples. Personal information of the producers, geographical locations, biosecurity measures and description of the poultry farms were recorded. The total number of poultry farms in Al-Jabal Al-Gharbi Region is 461 farms distributed in 13 cities. Out of these, 102 broiler farms and one broiler breeder farm (10 houses) which were in operation during team visit were included in this study. Following collection of blood, sera were separated and tested by Enzyme-linked immunosorbent assay (ELISA) for the presence of antibodies against Mycoplasma (General antigen for M. gallisepticum and M. synoviae). The seroprevalence of Mycoplasma in the region was 28% (29 poultry farms out of 103 were infected). About 50% (23 out of 47) of poultry farms located in Garian city were infected with Mycoplasma and one significant cluster of Mycoplasma infection in the city was identified. Low level of biosecurity was found in poultry farms of the region. Out of the 103 farms included, 63% of poultry houses has a ground of soil and 44% of them has uncoated walls which may influence the proper cleaning and disinfection. Almost 100% of the farms are at risk of exposure to diseases transmitted by wild birds such as avian influenza and Newcastle disease due to absence of wild birds control program. Although, 81% of the farms have entry restrictions, only 20% have disinfectants at entry which increase the risk of exposure to pathogens. The results of this study highlight the weakness points of biosecurity

  4. Biosecurity and geospatial analysis of mycoplasma infections in poultry farms at Al-Jabal Al-Gharbi region of Libya.

    PubMed

    Kammon, Abdulwahab; Mulatti, Paolo; Lorenzetto, Monica; Ferre, Nicola; Sharif, Monier; Eldaghayes, Ibrahim; Dayhum, Abdunaser

    2017-01-01

    Geospatial database of farm locations and biosecurity measures are essential to control disease outbreaks. A study was conducted to establish geospatial database on poultry farms in Al-Jabal Al-Gharbi region of Libya, to evaluate the biosecurity level of each farm and to determine the seroprevalence of mycoplasma and its relation to biosecurity level. A field team of 7 Veterinarians belongs to the National Center of Animal Health was assigned for data recording and collection of blood samples. Personal information of the producers, geographical locations, biosecurity measures and description of the poultry farms were recorded. The total number of poultry farms in Al-Jabal Al-Gharbi Region is 461 farms distributed in 13 cities. Out of these, 102 broiler farms and one broiler breeder farm (10 houses) which were in operation during team visit were included in this study. Following collection of blood, sera were separated and tested by Enzyme-linked immunosorbent assay (ELISA) for the presence of antibodies against Mycoplasma (General antigen for M. gallisepticum and M. synoviae). The seroprevalence of Mycoplasma in the region was 28% (29 poultry farms out of 103 were infected). About 50% (23 out of 47) of poultry farms located in Garian city were infected with Mycoplasma and one significant cluster of Mycoplasma infection in the city was identified. Low level of biosecurity was found in poultry farms of the region. Out of the 103 farms included, 63% of poultry houses has a ground of soil and 44% of them has uncoated walls which may influence the proper cleaning and disinfection. Almost 100% of the farms are at risk of exposure to diseases transmitted by wild birds such as avian influenza and Newcastle disease due to absence of wild birds control program. Although, 81% of the farms have entry restrictions, only 20% have disinfectants at entry which increase the risk of exposure to pathogens. The results of this study highlight the weakness points of biosecurity

  5. Enumeration, isolation, and species identification of mycoplasmas in saliva sampled from the normal and pathological human oral cavity and antibody response to an oral mycoplasma (Mycoplasma salivarium).

    PubMed

    Watanabe, T; Matsuura, M; Seto, K

    1986-06-01

    Saliva samples collected from 393 subjects with and without oral diseases were examined for concentrations of mycoplasmas and Mycoplasma species. Mycoplasmas were isolated from 383 (97%) of the 393 subjects. Viable counts ranged from zero to 7.6 X 10(7) CFU/ml (median, 6.9 X 10(4)) and were significantly (P less than 0.01) higher in diseased subjects, except for those with arthrosis temporomandibularis, than in controls. Of 1,400 isolates, 897 (64%), 442 (30%), and 8 (1%) were identified as Mycoplasma salivarium, M. orale, and M. hominis, respectively, and the remaining 73 isolates (5%) were unidentifiable. More than two-thirds of the isolates from diseased subjects versus only half from controls were identified as M. salivarium. In diseased subjects other than those with ostitis (especially those with arthrosis temporomandibularis), the incidence of M. salivarium was higher than that of M. orale, whereas the former occurred about as frequently as the latter in the controls. Antibodies to M. salivarium were also measured in sera from some subjects by the metabolism inhibition test. Sera with metabolism inhibition titers of 16 or greater were rated positive. There was no significant difference in the prevalence of antibodies between diseased subjects (60%) and controls (40%), but the mean titers (97 to 220) of all positive sera from diseased subjects were two to four times those for sera from controls. In addition, a fourfold or greater rise or fall of antibody titers to the organism was shown in paired sera from some subjects. On the basis of these results, M. salivarium was strongly suggested to participate etiologically in some cases of oral infection.

  6. Enumeration, isolation, and species identification of mycoplasmas in saliva sampled from the normal and pathological human oral cavity and antibody response to an oral mycoplasma (Mycoplasma salivarium).

    PubMed Central

    Watanabe, T; Matsuura, M; Seto, K

    1986-01-01

    Saliva samples collected from 393 subjects with and without oral diseases were examined for concentrations of mycoplasmas and Mycoplasma species. Mycoplasmas were isolated from 383 (97%) of the 393 subjects. Viable counts ranged from zero to 7.6 X 10(7) CFU/ml (median, 6.9 X 10(4)) and were significantly (P less than 0.01) higher in diseased subjects, except for those with arthrosis temporomandibularis, than in controls. Of 1,400 isolates, 897 (64%), 442 (30%), and 8 (1%) were identified as Mycoplasma salivarium, M. orale, and M. hominis, respectively, and the remaining 73 isolates (5%) were unidentifiable. More than two-thirds of the isolates from diseased subjects versus only half from controls were identified as M. salivarium. In diseased subjects other than those with ostitis (especially those with arthrosis temporomandibularis), the incidence of M. salivarium was higher than that of M. orale, whereas the former occurred about as frequently as the latter in the controls. Antibodies to M. salivarium were also measured in sera from some subjects by the metabolism inhibition test. Sera with metabolism inhibition titers of 16 or greater were rated positive. There was no significant difference in the prevalence of antibodies between diseased subjects (60%) and controls (40%), but the mean titers (97 to 220) of all positive sera from diseased subjects were two to four times those for sera from controls. In addition, a fourfold or greater rise or fall of antibody titers to the organism was shown in paired sera from some subjects. On the basis of these results, M. salivarium was strongly suggested to participate etiologically in some cases of oral infection. PMID:3711294

  7. Transcriptome Changes in Mycoplasma hyopneumoniae during Infection▿ †

    PubMed Central

    Madsen, Melissa L.; Puttamreddy, Supraja; Thacker, Eileen L.; Carruthers, Michael D.; Minion, F. Chris

    2008-01-01

    Mycoplasma hyopneumoniae causes swine pneumonia and contributes significantly to the porcine respiratory disease complex. The mechanisms of pathogenesis are difficult to address, since there is a lack of genetic tools, but microarrays are available and can be used to study transcriptional changes that occur during disease as a way to identify important virulence-related genes. Mycoplasmas were collected from bronchial alveolar lavage samples and compared to broth-grown cells using microarrays. Bronchial alveolar lavage was performed on pigs 28 days postinfection, and mycoplasmas were isolated by differential centrifugation. Mycoplasma RNA-enriched preparations were then obtained from total RNA by subtracting eucaryotic ribosomal and messenger RNAs. Labeled cDNAs were generated with mycoplasma open reading frame-specific primers. Nine biological replicates were analyzed. During lung infection, our analysis indicated that 79 M. hyopneumoniae genes were differentially expressed (P < 0.01), at a false-discovery rate of <2.7%. Of the down-regulated genes, 28 of 46 (61%) lacked an assigned function, in comparison to 21 of 33 (63%) of up-regulated genes. Four down-regulated genes and two up-regulated genes encoded putative lipoproteins. secA (mhp295) (P = 0.003) and two glycerol transport permease genes (potA [mhp380; P = 0.006] and ugpA [mhp381; P = 0.003]) were up-regulated in vivo. Elongation factor EF-G (fusA [mhp083]) (P = 0.002), RNA polymerase beta chain (rpoC [mhp635]) (P = 0.003), adenylate kinase (adk [mhp208]) (P = 0.001), prolyl aminoacyl tRNA synthetase (proS [mhp397]) (P = 0.009), and cysteinyl-tRNA synthetase (cysS [mhp661]) (P < 0.001) were down-regulated in vivo. PMID:18070898

  8. Remarkable increase in fluoroquinolone-resistant Mycoplasma genitalium in Japan.

    PubMed

    Kikuchi, Mina; Ito, Shin; Yasuda, Mitsuru; Tsuchiya, Tomohiro; Hatazaki, Kyoko; Takanashi, Masaki; Ezaki, Takayuki; Deguchi, Takashi

    2014-09-01

    We determined the prevalence of macrolide and fluoroquinolone resistance-associated mutations in Mycoplasma genitalium DNA specimens from men with non-gonococcal urethritis (NGU) and analysed their effects on antibiotic treatments of M. genitalium infections. In this retrospective study, we examined antibiotic resistance-associated mutations in the 23S rRNA, gyrA and parC genes of M. genitalium and the association of the mutations with microbiological outcomes of antibiotic treatments in men with M. genitalium-positive NGU. No macrolide resistance-associated mutations in the 23S rRNA gene were observed in 27 M. genitalium DNA specimens in 2011 and in 24 in 2012. However, 5 of 17 in 2013 had 23S rRNA mutations. Three of 15 in 2011, 6 of 19 in 2012 and 8 of 17 in 2013 had fluoroquinolone resistance-associated alterations in ParC. Three in 2013 had both the antibiotic resistance-associated alterations coincidentally. In two men with M. genitalium harbouring 23S rRNA mutations, the mycoplasma persisted after treatment with a regimen of 2 g of extended-release azithromycin (AZM-SR) once daily for 1 day. All nine men with mycoplasma harbouring ParC alterations were microbiologically cured with a regimen of 100 mg of sitafloxacin twice daily for 7 days. Macrolide- or fluoroquinolone-resistant M. genitalium appears to be increasing, and the increase in fluoroquinolone-resistant mycoplasmas is especially remarkable in Japan. Mycoplasmas harbouring 23S rRNA mutations would be resistant to the AZM-SR regimen, but those harbouring ParC alterations would still be susceptible to the sitafloxacin regimen. © The Author 2014. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  9. Discrimination between Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capricolum using PCR-RFLP and PCR.

    PubMed

    Cillara, Grazia; Manca, Maria Giovanna; Longheu, Carla; Tola, Sebastiana

    2015-09-01

    In this study, the dihydrolipoyl dehydrogenase (lpdA) gene was used to distinguish Mycoplasma mycoides subsp. capri (Mmc) from Mycoplasma capricolum subsp. capricolum (Mcc), two of four Mycoplasma species that cause contagious agalactia in sheep and goats. After alignment of nucleotide sequences of both species, specific primer sets were designed from unchanging and variable gene segments. The first primer set LPD-C1-F/LPD-C1-R was used to amplify a 911 bp fragment that was subsequently co-digested with FastDigest PstI, SspI, EcoRI and ClaI enzymes. The PCR-RFLP profiles differentiated the two mycoplasma species. The second primer set was used to distinguish Mmc from Mcc by single tube PCR. Both methods were further applied to identify 54 isolates collected from dairy herds from different provinces in Sardinia. The results of this study showed that PCR-RFLP and PCR could be used in routine diagnosis for rapid and specific simultaneous discrimination of Mmc and Mcc.

  10. What are mycoplasmas - The relationship of tempo and mode in bacterial evolution

    NASA Technical Reports Server (NTRS)

    Woese, C. R.; Stackebrand, E.; Ludwig, W.

    1985-01-01

    In phenotype the mycoplasmas are very different from ordinary bacteria. However, genotypically (i.e., phylogenetically) they are not. On the basis of ribosomal RNA homologies the mycoplasmas belong with the clostridia, and indeed have specific clostridial relatives. Mycoplasmas are, however, unlike almost all other bacteria in the evolutionary characteristics of their ribosomal RNAs. These RNAs contain relatively few of the highly conserved oligonucleotide sequences characteristic of normal eubacterial ribosomal RNAs. This is interpreted to be a reflection of an elevated mutation rate in mycoplasma lines of descent. A general consequence of this would be that the variation associated with a mycoplasma population is augmented both in number and kind, which in turn would lead to an unusual evolutionary course, one unique in all respects. Mycoplasmas, then, are actually tachytelic bacteria. The unusual evolutionary characteristics of their ribosomal RNAs are the imprints of their rapid evolution.

  11. What are mycoplasmas: the relationship of tempo and mode in bacterial evolution

    NASA Technical Reports Server (NTRS)

    Woese, C. R.; Stackebrandt, E.; Ludwig, W.

    1984-01-01

    In phenotype the mycoplasmas are very different from ordinary bacteria. However, genotypically (i.e., phylogenetically) they are not. On the basis of ribosomal RNA homologies the mycoplasmas belong with the clostridia, and indeed have specific clostridial relatives. Mycoplasmas are, however, unlike almost all other bacteria in the evolutionary characteristics of their ribosomal RNAs. These RNAs contain relatively few of the highly conserved oligonucleotide sequences characteristic of normal eubacterial ribosomal RNAs. This is interpreted to be a reflection of an elevated mutation rate in mycoplasma lines of descent. A general consequence of this would be that the variation associated with a mycoplasma population is augmented both in number and kind, which in turn would lead to an unusual evolutionary course, one unique in all respects. Mycoplasmas, then, are actually tachytelic bacteria. The unusual evolutionary characteristics of their ribosomal RNAs are the imprints of their rapid evolution.

  12. Novel hemotropic Mycoplasma species in white-tailed deer (Odocoileus virginianus).

    PubMed

    Maggi, Ricardo G; Chitwood, M Colter; Kennedy-Stoskopf, Suzanne; DePerno, Christopher S

    2013-12-01

    Globally, hemotropic Mycoplasma spp. are emerging or re-emerging zoonotic pathogens that affect livestock, wildlife, companion animals, and humans, potentially causing serious and economically important disease problems. Little is known about hemotropic Mycoplasma spp. prevalence, host-specificity, or route of transmission in most species, including wildlife. DNA amplification by PCR targeting the 16SrRNA and the RNaseP genes was used to establish the presence and prevalence of hemotropic Mycoplasma spp. in a white-tailed deer (O. virginianus) population in eastern North Carolina. Sixty-five deer (89%) tested positive for hemotropic Mycoplasma spp. where sequence analysis of the 16SsRNA and the RNaseP genes indicated the presence of at least three distinct species. This study represents the first detection of three distinct hemotropic Mycoplasma species in white-tailed deer and the first report of two novel hemotropic Mycoplasma species. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Infection with hemotropic Mycoplasma species in patients with or without extensive arthropod or animal contact.

    PubMed

    Maggi, Ricardo G; Compton, Sarah M; Trull, Chelsea L; Mascarelli, Patricia E; Mozayeni, B Robert; Breitschwerdt, Edward B

    2013-10-01

    PCR amplification targeting the 16S rRNA gene was used to test individuals with and without extensive arthropod and animal contact for the possibility of hemotropic mycoplasma infection. The prevalence of hemotropic mycoplasma infection (4.7%) was significantly greater in previously reported cohorts of veterinarians, veterinary technicians, spouses of veterinary professionals, and others with extensive arthropod exposure and/or frequent animal contact than in a previously reported cohort of patients examined by a rheumatologist because of chronic joint pain or evidence of small-vessel disease (0.7%). Based upon DNA sequence analysis, a Mycoplasma ovis-like species was the most prevalent organism detected; however, infection with "Candidatus Mycoplasma haematoparvum" and a potentially novel, but incompletely characterized, hemotropic Mycoplasma species was also documented. Historical exposure to animals and arthropod vectors that can harbor hemotropic Mycoplasma spp. should be considered during epidemiological investigations and in the evaluation of individual patients.

  14. What are mycoplasmas: the relationship of tempo and mode in bacterial evolution.

    PubMed

    Woese, C R; Stackebrandt, E; Ludwig, W

    In phenotype the mycoplasmas are very different from ordinary bacteria. However, genotypically (i.e., phylogenetically) they are not. On the basis of ribosomal RNA homologies the mycoplasmas belong with the clostridia, and indeed have specific clostridial relatives. Mycoplasmas are, however, unlike almost all other bacteria in the evolutionary characteristics of their ribosomal RNAs. These RNAs contain relatively few of the highly conserved oligonucleotide sequences characteristic of normal eubacterial ribosomal RNAs. This is interpreted to be a reflection of an elevated mutation rate in mycoplasma lines of descent. A general consequence of this would be that the variation associated with a mycoplasma population is augmented both in number and kind, which in turn would lead to an unusual evolutionary course, one unique in all respects. Mycoplasmas, then, are actually tachytelic bacteria. The unusual evolutionary characteristics of their ribosomal RNAs are the imprints of their rapid evolution.

  15. What are mycoplasmas - The relationship of tempo and mode in bacterial evolution

    NASA Technical Reports Server (NTRS)

    Woese, C. R.; Stackebrand, E.; Ludwig, W.

    1985-01-01

    In phenotype the mycoplasmas are very different from ordinary bacteria. However, genotypically (i.e., phylogenetically) they are not. On the basis of ribosomal RNA homologies the mycoplasmas belong with the clostridia, and indeed have specific clostridial relatives. Mycoplasmas are, however, unlike almost all other bacteria in the evolutionary characteristics of their ribosomal RNAs. These RNAs contain relatively few of the highly conserved oligonucleotide sequences characteristic of normal eubacterial ribosomal RNAs. This is interpreted to be a reflection of an elevated mutation rate in mycoplasma lines of descent. A general consequence of this would be that the variation associated with a mycoplasma population is augmented both in number and kind, which in turn would lead to an unusual evolutionary course, one unique in all respects. Mycoplasmas, then, are actually tachytelic bacteria. The unusual evolutionary characteristics of their ribosomal RNAs are the imprints of their rapid evolution.

  16. Identification and purification of arginine deiminase that originated from Mycoplasma arginini.

    PubMed Central

    Sugimura, K; Fukuda, S; Wada, Y; Taniai, M; Suzuki, M; Kimura, T; Ohno, T; Yamamoto, K; Azuma, I

    1990-01-01

    A lymphocyte blastogenesis inhibitory factor, (LBIF), was purified from the culture supernatant of human histiocytic lymphoma U937 by fast protein liquid chromatography. In this study, we demonstrated, first, that LBIF originated from a mycoplasma, Mycoplasma arginini, infecting U937 cells, and second, that LBIF bore the arginine deiminase activity. The implication of in vivo immunosuppression induced by arginine-utilizing mycoplasma species is discussed. Images PMID:2370103

  17. Distribution and diversity of mycoplasma plasmids: lessons from cryptic genetic elements

    PubMed Central

    2012-01-01

    Background The evolution of mycoplasmas from a common ancestor with Firmicutes has been characterized not only by genome down-sizing but also by horizontal gene transfer between mycoplasma species sharing a common host. The mechanisms of these gene transfers remain unclear because our knowledge of the mycoplasma mobile genetic elements is limited. In particular, only a few plasmids have been described within the Mycoplasma genus. Results We have shown that several species of ruminant mycoplasmas carry plasmids that are members of a large family of elements and replicate via a rolling-circle mechanism. All plasmids were isolated from species that either belonged or were closely related to the Mycoplasma mycoides cluster; none was from the Mycoplasma bovis-Mycoplasma agalactiae group. Twenty one plasmids were completely sequenced, named and compared with each other and with the five mycoplasma plasmids previously reported. All plasmids share similar size and genetic organization, and present a mosaic structure. A peculiar case is that of the plasmid pMyBK1 from M. yeatsii; it is larger in size and is predicted to be mobilizable. Its origin of replication and replication protein were identified. In addition, pMyBK1 derivatives were shown to replicate in various species of the M. mycoides cluster, and therefore hold considerable promise for developing gene vectors. The phylogenetic analysis of these plasmids confirms the uniqueness of pMyBK1 and indicates that the other mycoplasma plasmids cluster together, apart from the related replicons found in phytoplasmas and in species of the clade Firmicutes. Conclusions Our results unraveled a totally new picture of mycoplasma plasmids. Although they probably play a limited role in the gene exchanges that participate in mycoplasma evolution, they are abundant in some species. Evidence for the occurrence of frequent genetic recombination strongly suggests they are transmitted between species sharing a common host or niche. PMID

  18. The History of Mycoplasma pneumoniae Pneumonia

    PubMed Central

    Saraya, Takeshi

    2016-01-01

    Pinehurst transmission experiments resulted in a lapse of 20 years before acceptance of the Eaton agent as Mycoplasma pneumoniae. This review describes the history of M. pneumoniae pneumonia with a special focus on the recognition between the 1930 and 1960s of the Eaton agent as the infectious cause. PMID:27047477

  19. In Vitro Antibacterial Activity of AZD0914 against Human Mycoplasmas and Ureaplasmas

    PubMed Central

    Crabb, Donna M.; Duffy, Lynn B.; Huband, Michael D.

    2015-01-01

    In this study, susceptibilities were determined for AZD0914, a spiropyrimidinetrione DNA gyrase inhibitor, azithromycin, doxycycline, and levofloxacin against Mycoplasma and Ureaplasma species. The activity of AZD0914 was comparable to that of levofloxacin and doxycycline against Mycoplasma genitalium and Mycoplasma pneumoniae. The AZD0914 MIC90 against Mycoplasma hominis was 8-fold greater than that for levofloxacin. The AZD0914 MIC90 against Ureaplasma species was 4-fold less than that for azithromycin and 8-fold less than that for levofloxacin and doxycycline. PMID:25824220

  20. Mycoplasma detection and isolation from one-humped camels (Camelus dromedarius).

    PubMed

    Mederos-Iriarte, Lidia E; Poveda, José B; Poveda, Carlos G; Vega-Orellana, Orestes M; Gutiérrez, Carlos; Corbera, Juan A; Ramírez, Ana S

    2014-10-01

    In scientific literature, a small amount of information is found concerning mycoplasmosis in camel species. Mycoplasma (M.) arginini, Acholeplasma (A.) laidlawii, and Acholeplasma oculi have been reported to be isolated from these host species. Serologically positive results have been reported for Mycoplasma mycoides subsp. mycoides SC type, Mycoplasma capricolum subsp. capripneumoniae, and M. mycoides subsp. capri. The aims of this study were to detect, isolate, and identify mycoplasmas from camels (Camelus dromedarius). Initially, saliva and ear smears plus conjunctival and vaginal secretions were taken from five female animals, but only conjunctival secretions in three male animals, all belonging to the same farm. An unknown mycoplasma was isolated from one of the vagina samples. Additionally, another unknown and uncultured mycoplasma was detected with molecular biology in the same sample. In the second stage, 23 vaginal secretions were taken from the same farm plus another secretion from a different one. Ten isolates of the same unknown and previously isolated mycoplasma were detected, nine of them recovered from the vagina of female camels. Some mycoplasmas have been related to reproductive disorders; however, there is no evidence that the isolated mycoplasmas are related to such disorders.

  1. Comparison of the prevalence of Mycoplasma species in dogs with and without respiratory disease.

    PubMed

    Schulz, Bianka S; Raufeisen, Katharina; Weber, Karin; Laberke, Siija; Hartmann, Katrin

    2015-01-01

    Aim of the study was to investigate the prevalence of Mycoplasma species in dogs with and without signs of respiratory disease. Bronchoalveolarlavage fluid (BALF) and pharyngeal swabs were collected from 29 dogs with respiratory diseases (RD) and 16 dogs without signs of RD that were euthanised because of other diseases. Samples were tested for Mycoplasma species by PCR and culture, and sequencing was performed in Mycoplasma species-positive BALF samples. Pharyngeal swabs were positive for Mycoplasma species by PCR in 91.7% of dogs with RD and 86.7% of dogs without signs of RD (p = 1.000); BALF samples were PCR-positive in 37.9% of dogs with RD and 18.8% of dogs without signs of RD (p = 0.194) Mycoplasmo culture of BALF was positive in 28.6% of dogs with RD and in 18.8% without signs of RD (p = 0.730). When culture and PCR were compared, there was no significant difference in the detection rate of Mycoplasma species (p = 0.658) Sequencing detected different Mycoplasma species. Out of these, however, Mycoplasma cynos was isolated from four dogs with RD. There is no significant difference in the prevalence of Mycoplasma species between dogs with RD and dogs without evidence of RD; however, Mycoplasma cynos seems to be associated with respiratory disease.

  2. Detection and Antibiotic Treatment of Mycoplasma arginini Contamination in a Mouse Epithelial Cell Line Restore Normal Cell Physiology

    PubMed Central

    Resnick, Andrew

    2014-01-01

    Mycoplasma contamination of cultured cell lines is difficult to detect by routine observation. Infected cells can display normal morphology and the slow growth rate of mycoplasma can delay detection for extended periods of time, compromising experimental results. Positive identification of mycoplasma typically requires cells to be either fixed and stained for DNA or processed with PCR. We present a method to detect mycoplasma using live-cell optical microscopy typically used for routine observation of cell cultures. Images of untreated mycoplasma-infected epithelial cells alongside images of infected cells treated with Plasmocin, a commercially available antibiotic targeted to mycoplasma, are shown. We found that optical imaging is an effective screening tool for detection of mycoplasma contamination. Importantly, we found that cells regained normal function after the contamination was cleared. In conclusion, we present a technique to diagnose probable mycoplasma infections in live cultures without fixation, resulting in faster response times and decreased loss of cell material. PMID:24772428

  3. Generation of a monoclonal antibody against Mycoplasma spp. following accidental contamination during production of a monoclonal antibody against Lawsonia intracellularis.

    PubMed

    Hwang, Jeong-Min; Lee, Ji-Hye; Yeh, Jung-Yong

    2012-03-01

    This report describes Mycoplasma contamination of Lawsonia intracellularis cultures that led to the unintended acquisition of a monoclonal antibody against Mycoplasma spp. during the attempted generation of a monoclonal antibody against L. intracellularis.

  4. Detection and antibiotic treatment of Mycoplasma arginini contamination in a mouse epithelial cell line restore normal cell physiology.

    PubMed

    Boslett, Brianna; Nag, Subhra; Resnick, Andrew

    2014-01-01

    Mycoplasma contamination of cultured cell lines is difficult to detect by routine observation. Infected cells can display normal morphology and the slow growth rate of mycoplasma can delay detection for extended periods of time, compromising experimental results. Positive identification of mycoplasma typically requires cells to be either fixed and stained for DNA or processed with PCR. We present a method to detect mycoplasma using live-cell optical microscopy typically used for routine observation of cell cultures. Images of untreated mycoplasma-infected epithelial cells alongside images of infected cells treated with Plasmocin, a commercially available antibiotic targeted to mycoplasma, are shown. We found that optical imaging is an effective screening tool for detection of mycoplasma contamination. Importantly, we found that cells regained normal function after the contamination was cleared. In conclusion, we present a technique to diagnose probable mycoplasma infections in live cultures without fixation, resulting in faster response times and decreased loss of cell material.

  5. Effect of pH on Human Mycoplasma Strains

    PubMed Central

    Shepard, Maurice C.; Lunceford, Carl D.

    1965-01-01

    Shepard, Maurice C. (U.S. Naval Medical Field Research Laboratory, Camp Lejeune, N.C.), and Carl D. Lunceford. Effect of pH on human Mycoplasma strains. J. Bacteriol. 89:265–270. 1965.—The optimal reaction of culture media for the cultivation of T-strain Mycoplasma of human origin was investigated. By use of a recently modified tryptic digest medium, the optimal reaction in either agar or fluid medium was found to be pH 6.0. In contrast, human classic (large-colony) Mycoplasma could be cultivated in agar or fluid medium over a rather broad pH range, and the influence of the reaction of the medium appeared to be primarily species-dependent. M. salivarium, for example, grew best in agar from pH 5.5 through 6.5. M. pneumoniae (Easton's agent) yielded largest colony numbers in agar and highest titers in broth at pH 8.0. In the case of T-strain Mycoplasma, both maximal colony numbers in agar and highest titers in fluid media were achieved at a reaction of pH 6.0. In addition, largest colony size of T-strain Mycoplasma was also achieved in agar at pH 6.0, and averaged 50 to 100% larger than that obtained by cultivation at pH 8.0 with the same medium. Although T-strains will develop in agar media over a pH range of from 5.0 through 10.0, the extremely small colony size and poor staining properties resulting from growth in an alkaline medium make their recognition in agar cultures difficult. Aerobic cultivation of T-strains was first achieved in agar adjusted to pH 5.5 to 6.0. In fluid medium, multiplication of T-strains occurred only within the limits of pH 5.0 through 8.0, with highest titers being reached at pH 6.0. Greater attention to the reaction of complete Mycoplasma media is stressed. PMID:14255688

  6. Collaborative study report: evaluation of the ATCC experimental mycoplasma reference strains panel prepared for comparison of NAT-based and conventional mycoplasma detection methods.

    PubMed

    Dabrazhynetskaya, Alena; Volokhov, Dmitriy V; Lin, Tsai-Lien; Beck, Brian; Gupta, Rajesh K; Chizhikov, Vladimir

    2013-11-01

    The main goal of this collaborative study was to evaluate the experimental panel of cryopreserved mycoplasma reference strains recently prepared by the American Type Culture Collection (ATCC(®)) in order to assess the viability and dispersion of cells in the mycoplasma stocks by measuring the ratio between the number of genomic copies (GC) and the number of colony forming units (CFU) in the reference preparations. The employment of microbial reference cultures with low GC/CFU ratios is critical for unbiased and reliable comparison of mycoplasma testing methods based on different methodological approaches, i.e., Nucleic Acid Testing (NAT) and compendial culture-based techniques. The experimental panel included ten different mycoplasma species known to represent potential human and animal pathogens as well as common contaminants of mammalian and avian cell substrates used in research, development, and manufacture of biological products. Fifteen laboratories with expertise in field of mycoplasma titration and quantification of mycoplasmal genomic DNA participated in the study conducted from February to October of 2012. The results of this study demonstrated the feasibility of preparing highly viable and dispersed (possessing low GC/CFU ratios) frozen stocks of mycoplasma reference materials, required for reliable comparison of NAT-based and conventional mycoplasma detection methods.

  7. Detection of feline Mycoplasma species in cats with feline asthma and chronic bronchitis.

    PubMed

    Schulz, Bianka S; Richter, Petra; Weber, Karin; Mueller, Ralf S; Wess, Gerhard; Zenker, Isabella; Hartmann, Katrin

    2014-12-01

    Little is known about the aetiology of inflammatory lower airway disease in cats. The aim of this study was to investigate the role of Mycoplasma species in cats with feline asthma (FA) and chronic bronchitis (CB). The study population consisted of 17 cats with FA/CB, and 14 sick cats without clinical and historical signs of respiratory disease, which were euthanased for various other reasons. Nasal swabs, nasal lavage and bronchoalveolar lavage fluid (BALF) samples were taken from patients from both groups. Mycoplasma species culture with modified Hayflick agar and Mycoplasma polymerase chain reaction (PCR) were performed on all samples followed by sequencing of all Mycoplasma species-positive samples for differentiation of subspecies. PCR testing detected significantly more Mycoplasma species-positive BALF samples than Mycoplasma culture (P = 0.021). When cats with oropharyngeal contamination were excluded from comparison, the numbers of Mycoplasma species-positive BALF samples in the group with FA/CB (6/17) and the control group (4/9) were not significantly different (P = 0.6924). While all nasal samples of the cats with FA/CB were negative for Mycoplasma organisms, five samples in the control group (P = 0.041) were positive on PCR. Sequencing revealed Mycoplasma felis in all PCR-positive samples. Mycoplasma species can be detected in the lower airways of cats with FA/CB, as well as in the BALF of sick cats without respiratory signs. Further studies are warranted to investigate the possibility that Mycoplasma species represent commensals of the lower respiratory tract of cats.

  8. Isolation and Characterization of Mycoplasma sphenisci sp. nov. from the Choana of an Aquarium-Reared Jackass Penguin (Spheniscus demersus)

    PubMed Central

    Frasca, Salvatore; Weber, E. Scott; Urquhart, Heather; Liao, Xiaofen; Gladd, Martha; Cecchini, Katharine; Hudson, Paul; May, Meghan; Gast, Rebecca J.; Gorton, Timothy S.; Geary, Steven J.

    2005-01-01

    Strain UCMJ was isolated from the choana of a jackass penguin (Spheniscus demersus) with recurrent mucocaseous choanal discharge. Isolation of this mycoplasma expands the known range of species hosting mycoplasmas. The name Mycoplasma sphenisci sp. nov. is proposed for this new species, for which strain UCMJ is the type strain. PMID:15956436

  9. Studies into the prevalence of Mycoplasma species in small ruminants in Benue State, North-central Nigeria.

    PubMed

    Akwuobu, Chinedu A; Ayling, Roger D; Chah, Kennedy Foinkfu; Oboegbulem, Stephen I

    2014-08-01

    The indicative prevalence of respiratory Mycoplasma species in small ruminants (SR) was determined in North-central Nigeria. Nasal swabs from 172 sheep and 336 goats from the Northeast, Northwest and South Senatorial Districts of Benue State were examined. Initial Mycoplasma isolation used Mycoplasma culture techniques followed by digitonin sensitivity testing. Species identification was done using polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE). Overall, Mycoplasma organisms were isolated from 131 (25.8 %) of the 508 SR examined. Prevalence rates of 18.1 and 29.8 % were recorded for sheep and goats, respectively. A total of 135 isolates of Mycoplasma belonging to three different species were identified: Mycoplasma ovipneumoniae (127), Mycoplasma arginini (7) and Mycoplasma mycoides subspecies capri (1). More than one Mycoplasma species were detected in four (3.1 %) of the 131 confirmed Mycoplasma positive cultures. Mycoplasma was isolated from 16.2 and 29.1 % of animals with and without respiratory signs, respectively. The high isolation rate of mycoplasmas in apparently healthy and clinically sick sheep and goats in this study indicates a carrier status in these SR which may constitute a serious problem in disease control.

  10. Mycoplasma testudineum in free-ranging desert tortoises, Gopherus agassizii

    USGS Publications Warehouse

    Jacobson, Elliott R.; Berry, Kristin H.

    2012-01-01

    We performed clinico-pathological evaluations of 11 wild Agassiz's desert tortoises (Gopherus agassizii) from a translocation project in the central Mojave Desert, California, USA. Group 1 consisted of nine tortoises that were selected primarily due to serologic status, indicating exposure to Mycoplasma testudineum (seven) or both M. agassizii and M. testudineum (two), and secondarily due to clinical signs of upper respiratory tract disease (URTD). Group 2 consisted of two tortoises that were antibody-negative for Mycoplasma and had no clinical signs of URTD, but did have other signs of illness. Of the Group 1 tortoises, M. testudineum, but not M. agassizii, was amplified by polymerase chain reaction and DNA fingerprinted from two tortoises. Using light microscopy, mild to severe pathologic changes were observed in one or more histologic sections of either one or both nasal cavities of each tortoise in Group 1. Our findings support a causal relationship between M. testudineum and URTD in desert tortoises.

  11. Analysis of energy sources for Mycoplasma penetrans gliding motility.

    PubMed

    Jurkovic, Dominika A; Hughes, Michael R; Balish, Mitchell F

    2013-01-01

    Mycoplasma penetrans, a potential human pathogen found mainly in HIV-infected individuals, uses a tip structure for both adherence and gliding motility. To improve our understanding of the molecular mechanism of M. penetrans gliding motility, we used chemical inhibitors of energy sources associated with motility of other organisms to determine which of these is used by M. penetrans and also tested whether gliding speed responded to temperature and pH. Mycoplasma penetrans gliding motility was not eliminated in the presence of a proton motive force inhibitor, a sodium motive force inhibitor, or an agent that depletes cellular ATP. At near-neutral pH, gliding speed increased as temperature increased. The absence of a clear chemical energy source for gliding motility and a positive correlation between speed and temperature suggest that energy derived from heat provides the major source of power for the gliding motor of M. penetrans.

  12. Mycoplasmacidal activity of bovine milk for T-mycoplasmas.

    PubMed

    Brownlie, J; Howard, C J; Gourlay, R N

    1974-12-01

    Normal bovine milk and whey was mycoplasmacidal for 6 of the 13 strains of bovine T-mycoplasmas examined. The in vitro assay used also demonstrated no killing of the human, canine and simian T-mycoplasma strains after 4 hr. incubation. However, there appeared to be some cow-to-cow variation in possession of this activity, and following E. coli endotoxin stimulation of the mammary gland the activity was considerably reduced.Whey from three normal cows was fractionated on a Bio-Gel A 1.5 m. column and the mycoplasmacidal activity of the resulting five peaks assayed. Only the second peak, peak B, contained activity and was characterized as the only peak containing bovine IgA. The active component in whey, however, was found to be heat stable at 60 degrees C. for 60 minutes and to pass through a dialysis membrane. This is inconsistent with it being immunoglobulin.

  13. Detection of Mycoplasma pneumoniae by using the polymerase chain reaction.

    PubMed Central

    Bernet, C; Garret, M; de Barbeyrac, B; Bebear, C; Bonnet, J

    1989-01-01

    The polymerase chain reaction (PCR) technique was used to detect Mycoplasma pneumoniae. A specific DNA sequence for M. pneumoniae was selected from a genomic library, and two oligonucleotides were chosen in this sequence to give an amplified fragment of 144 base pairs. We show that DNA from different M. pneumoniae strains can be detected by PCR, with DNA from other Mycoplasma species giving negative results. Analysis of biological samples (throat swabs) obtained from hamsters that were experimentally infected with M. pneumoniae showed that PCR was more sensitive and reliable than conventional culture techniques for the detection of M. pneumoniae. Initial experiments on artificially seeded human bronchoalveolar lavages showed that PCR can be used to detect 10(2) to 10(3) organisms. Images PMID:2509513

  14. Phospholipids and Glycolipids of Sterol-requiring Mycoplasma

    PubMed Central

    Smith, Paul F.; Koostra, Walter L.

    1967-01-01

    The phospholipids of Mycoplasma hominis type 2 strain 07 are composed almost entirely of phosphatidyl glycerol. Traces of other glycerophospholipids may exist. No glycolipids are found. The phospholipids of Mycoplasma sp. avian strain J are composed of diphosphatidyl glycerol, which predominates in older cultures, a monoacyl glycerophosphoryl glycerophosphate, which may serve as a precursor of diphosphatidyl glycerol, and phosphatidyl glycerophosphate. This organism also contains cholesteryl glucoside and an unidentified glycolipid which appears to be similar to a monoglucosyl diglyceride. No turnover or radioisotope labeling of the phospholipids occurs during metabolism. This lack of turnover during growth is indicative of a structural role for these glycerophospholipids. A concomitant decrease of monoacyl glycerophosphoryl glycerophosphate and increase of diphosphatidyl glycerol occurs during growth. PMID:6025304

  15. Mycoplasma synoviae infection on Newcastle disease vaccination of chickens

    PubMed Central

    de Cássia Figueira Silva, Rita; do Nascimento, Elmiro Rosendo; de Almeida Pereira, Virgínia Léo; Barreto, Maria Lúcia; do Nascimento, Maria da Graça Fichel

    2008-01-01

    Newcastle disease is characterized by respiratory manifestations in association with nervous and/or digestive symptoms. Its prevention is done by vaccination with live attenuated (lentogenic strains) and/or killed vaccines. The lentogenic strains can lead to strong post-vaccination reaction, principally due to the presence of other pathogenic agents. Among them, Mycoplasma synoviae is worldwide important, mainly in Brazil. The dissemination of this agent in poultry flocks has been achieved due to difficulties in diagnosis and disease reproduction, virulence variations among different M.synoviae strains, and attribution of typical M.synoviae disease manifestation to other disease agents. This experimental study in SPF chicks (Gallus gallus), previously infected by M.synoviae and thereafter vaccinated against Newcastle disease, was done with the objective of evaluating M.synoviae pathogenicity through assessment of post-vaccinal respiratory reactions and serologic responses to Newcastle disease virus vaccine in the absence of environmental factors. A total of 86 three days old chicks were used, being 57 infected by eye and nostril drop, with chicken activated M. synoviae strain WVU 1853. Seven days later, 21 mycoplasma infected birds plus 29 not mycoplasma infected ones were vaccinated against Newcastle disease. As results, the not infected and vaccinated birds yielded, significantly, higher and longer lasting serologic responses to Newcastle disease vaccine virus than those infected and vaccinated. Similarly, the infected and vaccinated birds yielded lower serologic reactions to M.synoviae than those only mycoplasma infected. No post-vaccinal respiratory reaction was observed in the vaccinated birds. PMID:24031234

  16. In vitro activity of chlorhydroxyquinoline against mycoplasma species.

    PubMed

    Cosgrove, R F; Baines, S

    1978-03-01

    The in vitro activities of 5-chloro-8-hydroxyquinoline (CHQ) against single strains of 12 different species of mycoplasma and the impacts of repeated exposure of these strains to CHQ on their susceptibility to this agent have been studied. On initial exposure, the minimal inhibitory concentrations for these strains ranged from 0.24 to 1.92 micrograms of CHQ per ml of test medium; activities remained unchanged during 10 serial transfers in CHQ-containing medium.

  17. In vitro activity of chlorhydroxyquinoline against mycoplasma species.

    PubMed Central

    Cosgrove, R F; Baines, S

    1978-01-01

    The in vitro activities of 5-chloro-8-hydroxyquinoline (CHQ) against single strains of 12 different species of mycoplasma and the impacts of repeated exposure of these strains to CHQ on their susceptibility to this agent have been studied. On initial exposure, the minimal inhibitory concentrations for these strains ranged from 0.24 to 1.92 micrograms of CHQ per ml of test medium; activities remained unchanged during 10 serial transfers in CHQ-containing medium. PMID:263891

  18. A change in the genetic code in Mycoplasma capricolum

    NASA Technical Reports Server (NTRS)

    Jukes, T. H.

    1985-01-01

    Mycoplasma capricolum was previously found to use UGA instead of UGG as its codon for tryptophan and to contain 75 percent A + T in its DNA. The codon change could have been due to mutational pressure to replace C + G by A + T, resulting in the replacement of UGA stop codons by UAA, change of the anticodon in tryptophan tRNA from CCA to UCA, and replacement of UGG tryptophan codons by UGA. None of these changes should have been deleterious.

  19. A change in the genetic code in Mycoplasma capricolum

    NASA Technical Reports Server (NTRS)

    Jukes, T. H.

    1985-01-01

    Mycoplasma capricolum was previously found to use UGA instead of UGG as its codon for tryptophan and to contain 75 percent A + T in its DNA. The codon change could have been due to mutational pressure to replace C + G by A + T, resulting in the replacement of UGA stop codons by UAA, change of the anticodon in tryptophan tRNA from CCA to UCA, and replacement of UGG tryptophan codons by UGA. None of these changes should have been deleterious.

  20. Rhamnose Links Moonlighting Proteins to Membrane Phospholipid in Mycoplasmas

    PubMed Central

    Daubenspeck, James M.; Liu, Runhua; Dybvig, Kevin

    2016-01-01

    Many proteins that have a primary function as a cytoplasmic protein are known to have the ability to moonlight on the surface of nearly all organisms. An example is the glycolytic enzyme enolase, which can be found on the surface of many types of cells from bacteria to human. Surface enolase is not enzymatic because it is monomeric and oligomerization is required for glycolytic activity. It can bind various molecules and activate plasminogen. Enolase lacks a signal peptide and the mechanism by which it attaches to the surface is unknown. We found that treatment of whole cells of the murine pathogen Mycoplasma pulmonis with phospholipase D released enolase and other common moonlighting proteins. Glycostaining suggested that the released proteins were glycosylated. Cytoplasmic and membrane-bound enolase was isolated by immunoprecipitation. No post-translational modification was detected on cytoplasmic enolase, but membrane enolase was associated with lipid, phosphate and rhamnose. Treatment with phospholipase released the lipid and phosphate from enolase but not the rhamnose. The site of rhamnosylation was identified as a glutamine residue near the C-terminus of the protein. Rhamnose has been found in all species of mycoplasma examined but its function was previously unknown. Mycoplasmas are small bacteria with have no peptidoglycan, and rhamnose in these organisms is also not associated with polysaccharide. We suggest that rhamnose has a central role in anchoring proteins to the membrane by linkage to phospholipid, which may be a general mechanism for the membrane association of moonlighting proteins in mycoplasmas and perhaps other bacteria. PMID:27603308

  1. A serological investigation of Mycoplasma pneumoniae infection on the Witwatersrand.

    PubMed

    Joosting, A C; Harwin, R M; Coppin, A; Battaglia, P; van der Hoef, P

    1976-12-18

    Sera from patients with respiratory disease were examined for antibody to Mycoplasma pneumoniae by complement fixation test. During the study period of about 6 years, a 3-year cycle of infection was observed, which coincided with some epidemics in the UK and USA, suggesting the possibility of an approximately simultaneous world-wide spread. The epidemics lasted about 18 months each, during which the incidence of infection was over 10 times that of the interepidemic periods.

  2. Characterization of Free Exopolysaccharides Secreted by Mycoplasma mycoides Subsp. mycoides

    PubMed Central

    Bertin, Clothilde; Pau-Roblot, Corinne; Courtois, Josiane; Manso-Silván, Lucía; Thiaucourt, François; Tardy, Florence; Le Grand, Dominique; Poumarat, François; Gaurivaud, Patrice

    2013-01-01

    Contagious bovine pleuropneumonia is a severe respiratory disease of cattle that is caused by a bacterium of the Mycoplasma genus, namely Mycoplasma mycoides subsp. mycoides (Mmm). In the absence of classical virulence determinants, the pathogenicity of Mmm is thought to rely on intrinsic metabolic functions and specific components of the outer cell surface. One of these latter, the capsular polysaccharide galactan has been notably demonstrated to play a role in Mmm persistence and dissemination. The free exopolysaccharides (EPS), also produced by Mmm and shown to circulate in the blood stream of infected cattle, have received little attention so far. Indeed, their characterization has been hindered by the presence of polysaccharide contaminants in the complex mycoplasma culture medium. In this study, we developed a method to produce large quantities of EPS by transfer of mycoplasma cells from their complex broth to a chemically defined medium and subsequent purification. NMR analyses revealed that the purified, free EPS had an identical β(1−>6)-galactofuranosyl structure to that of capsular galactan. We then analyzed intraclonal Mmm variants that produce opaque/translucent colonies on agar. First, we demonstrated that colony opacity was related to the production of a capsule, as observed by electron microscopy. We then compared the EPS extracts and showed that the non-capsulated, translucent colony variants produced higher amounts of free EPS than the capsulated, opaque colony variants. This phenotypic variation was associated with an antigenic variation of a specific glucose phosphotransferase permease. Finally, we conducted in silico analyses of candidate polysaccharide biosynthetic pathways in order to decipher the potential link between glucose phosphotransferase permease activity and attachment/release of galactan. The co-existence of variants producing alternative forms of galactan (capsular versus free extracellular galactan) and associated with an

  3. Mycoplasma and associated bacteria isolated from ovine pink-eye.

    PubMed

    Langford, E V

    1971-01-01

    A mycoplasma was recovered from the untreated conjunctival membranes of nine sheep affected by Pink-eye. It was neither isolated from the conjunctiva of treated animals which were affected nor from the conjunctiva of normal animals either in contact or not in contact with affected animals. Bacteria found on normal conjunctival membranes were Neisseria ovis, Escherichia coli, Staphylococcus epidermididis, Streptococcus and Bacillus spp. Bacteria found in clinical cases of Pink-eye were N. ovis, E. coli, a Streptococcus and Pseudomonas spp.

  4. Hemotropic mycoplasma infection in wild black bears (Ursus thibetanus japonicus).

    PubMed

    Iso, Takehiro; Suzuki, Jin; Sasaoka, Fumina; Sashida, Hinako; Watanabe, Yusaku; Fujihara, Masatoshi; Nagai, Kazuya; Harasawa, Ryô

    2013-04-12

    This is the first report on Mycoplasma infection in wild bears. We report a novel hemotropic Mycoplasma (also called hemoplasma) detected in a free-ranging black bear (Ursus thibetanus japonicus) in Japan. We then used real-time PCR to look for hemoplasma DNA in blood samples collected from 15 bears and found that eight (53%) were positive. Among these eight PCR samples, seven showed a melting temperature of around 85.5°C, while the remaining one showed a single peak at 82.26°C. Almost the entire region of the 16S rRNA gene as well as the 16S-23S rRNA intergenic transcribed spacer (ITS) region from the sample that showed a melting temperature of 82.26°C was successfully amplified by means of end-point PCR. The nucleotide sequences of the 16S rRNA gene and the ITS region were then determined and compared with those of authentic Mycoplasma species. Our examinations revealed the presence of a novel hemoplasma in Japanese black bears. Copyright © 2012 Elsevier B.V. All rights reserved.

  5. An Emerging Mycoplasma Associated with Trichomoniasis, Vaginal Infection and Disease

    PubMed Central

    Fettweis, Jennifer M.; Serrano, Myrna G.; Huang, Bernice; Brooks, J. Paul; Glascock, Abigail L.; Sheth, Nihar U.; Strauss, Jerome F.; Jefferson, Kimberly K.; Buck, Gregory A.

    2014-01-01

    Humans are colonized by thousands of bacterial species, but it is difficult to assess the metabolic and pathogenic potential of the majority of these because they have yet to be cultured. Here, we characterize an uncultivated vaginal mycoplasma tightly associated with trichomoniasis that was previously known by its 16S rRNA sequence as “Mnola.” In this study, the mycoplasma was found almost exclusively in women infected with the sexually transmitted pathogen Trichomonas vaginalis, but rarely observed in women with no diagnosed disease. The genomes of four strains of this species were reconstructed using metagenome sequencing and assembly of DNA from four discrete mid-vaginal samples, one of which was obtained from a pregnant woman with trichomoniasis who delivered prematurely. These bacteria harbor several putative virulence factors and display unique metabolic strategies. Genes encoding proteins with high similarity to potential virulence factors include two collagenases, a hemolysin, an O-sialoglycoprotein endopeptidase and a feoB-type ferrous iron transport system. We propose the name “Candidatus Mycoplasma girerdii” for this potential new pathogen. PMID:25337710

  6. An emerging mycoplasma associated with trichomoniasis, vaginal infection and disease.

    PubMed

    Fettweis, Jennifer M; Serrano, Myrna G; Huang, Bernice; Brooks, J Paul; Glascock, Abigail L; Sheth, Nihar U; Strauss, Jerome F; Jefferson, Kimberly K; Buck, Gregory A

    2014-01-01

    Humans are colonized by thousands of bacterial species, but it is difficult to assess the metabolic and pathogenic potential of the majority of these because they have yet to be cultured. Here, we characterize an uncultivated vaginal mycoplasma tightly associated with trichomoniasis that was previously known by its 16S rRNA sequence as "Mnola." In this study, the mycoplasma was found almost exclusively in women infected with the sexually transmitted pathogen Trichomonas vaginalis, but rarely observed in women with no diagnosed disease. The genomes of four strains of this species were reconstructed using metagenome sequencing and assembly of DNA from four discrete mid-vaginal samples, one of which was obtained from a pregnant woman with trichomoniasis who delivered prematurely. These bacteria harbor several putative virulence factors and display unique metabolic strategies. Genes encoding proteins with high similarity to potential virulence factors include two collagenases, a hemolysin, an O-sialoglycoprotein endopeptidase and a feoB-type ferrous iron transport system. We propose the name "Candidatus Mycoplasma girerdii" for this potential new pathogen.

  7. Vaccines for Mycoplasma diseases in animals and man.

    PubMed

    Nicholas, R A J; Ayling, R D; McAuliffe, L

    2009-01-01

    Vaccines for important mycoplasma diseases, including contagious bovine and caprine pleuropneumonia, have been used for centuries, consisting mainly of infected tissue or fluids which are inoculated into sites at which the risk of severe infection is slight, such as the tail and bridge of the nose. Surprisingly, little progress has been made in developing safe, defined and protective alternatives, the vaccines today still consisting of mildly attenuated strains serially passaged in eggs or in culture. Ill-defined temperature-sensitive mutants are widely used for mycoplasmoses in poultry despite uncertainty about their mode of protection. Inactivated vaccines for enzootic pneumonia appear to have improved pig health worldwide, but disease reduction has been generally modest. Ironically, attempts to develop subunit preparations have often led to exacerbation of disease, particularly in human atypical pneumonia. Promising results have been seen in DNA vaccine technology, which has been applied to the development of mycoplasma vaccines for porcine enzootic pneumonia, but field trials still seem a long way off. No commercial vaccines exist for Mycoplasma bovis, despite evidence that this is a major cause of calf pneumonia, mastitis and arthritis.

  8. Acquired immunity to Mycoplasma pneumoniae. Pneumonia in hamsters.

    PubMed

    Hayatsu, E

    1978-01-01

    An inactivated Mycoplasma pneumoniae vaccine was prepared from a culture in a liquid medium supplemented with water extract of egg yolk. Vaccinated Syrian hamsters were exposed to virulent M. pneumoniae aerosol and were examined for the retention of mycoplasmas and for histopathological changes in the respiratory tracts. When a vaccine prepared with strain FH was administered intramuscularly or by inhalation in aerosol, no significant resistance was shown with respect to mycoplasma proliferation. An increased resistance, however, was observed when an aluminium phosphate-adsorbed vaccine, and when a plain vaccine (although to a lesser degree) prepared with hamster 24-passaged strain FH, was administered intramuscularly. Histopathologically, lung lesions were markedly suppressed in groups showing high resistance. A correlation between the serum antibody titer and the resistance to infection was observed. Hamsters which received a hyperimmune rabbit antiserum intracordally showed a high resistance to M. pneumoniae infection. The suppression of histopathological changes also coincided with high complement-fixing antibody titers of either actively or passively immunized hamster serum. The results suggest that humoral immunity plays an important role in resistance to M. pneumoniae pneumonia in hamsters.

  9. Comparative Analysis of Gene Content Evolution in Phytoplasmas and Mycoplasmas

    PubMed Central

    Lin, Chan-Pin; Kuo, Chih-Horng

    2012-01-01

    Phytoplasmas and mycoplasmas are two groups of important pathogens in the bacterial class Mollicutes. Because of their economical and clinical importance, these obligate pathogens have attracted much research attention. However, difficulties involved in the empirical study of these bacteria, particularly the fact that phytoplasmas have not yet been successfully cultivated outside of their hosts despite decades of attempts, have greatly hampered research progress. With the rapid advancements in genome sequencing, comparative genome analysis provides a new approach to facilitate our understanding of these bacteria. In this study, our main focus is to investigate the evolution of gene content in phytoplasmas, mycoplasmas, and their common ancestor. By using a phylogenetic framework for comparative analysis of 12 complete genome sequences, we characterized the putative gains and losses of genes in these obligate parasites. Our results demonstrated that the degradation of metabolic capacities in these bacteria has occurred predominantly in the common ancestor of Mollicutes, prior to the evolutionary split of phytoplasmas and mycoplasmas. Furthermore, we identified a list of genes that are acquired by the common ancestor of phytoplasmas and are conserved across all strains with complete genome sequences available. These genes include several putative effectors for the interactions with hosts and may be good candidates for future functional characterization. PMID:22479625

  10. Isolation of Mycoplasma genitalium strains from the male urethra.

    PubMed Central

    Jensen, J S; Hansen, H T; Lind, K

    1996-01-01

    Mycoplasma genitalium is a human mycoplasma species which, on the basis of detection by PCR, has been incriminated as a cause of nongonococcal urethritis. Previously, only two strains from the urogenital tract and five strains from extragenital sites have been isolated. We have developed a method for the isolation of this fastidious microbe. M. genitalium from PCR-positive urethral specimens was initially propagated in Vero cell cultures grown in serum-free medium supplemented with Ultroser HY serum substitute. Growth was monitored by PCR. The M. genitalium strains grown in cell cultures could subsequently be subcultured in modified Friis's FF broth medium. Several passages in broth medium were required before growth on agar medium was attained. A total of 11 urethral specimens positive for M. genitalium by PCR from male patients with urethritis were investigated. Six strains were adapted to growth in broth medium, and four of these strains were cloned. Three specimens were overgrown by other mycoplasmas during propagation in the cell cultures. In only two PCR-positive specimens was propagation of M. genitalium unsuccessful. The use of cell culture combined with PCR monitoring of mycoplasmal growth may prove to be more widely applicable for the isolation of other fastidious mollicutes. PMID:8789002

  11. Transcriptional Profiling of Mycoplasma hyopneumoniae during Heat Shock Using Microarrays†

    PubMed Central

    Madsen, Melissa L.; Nettleton, Dan; Thacker, Eileen L.; Edwards, Robert; Minion, F. Chris

    2006-01-01

    Bacterial pathogens undergo stress during host colonization and disease processes. These stresses result in changes in gene expression to compensate for potentially lethal environments developed in the host during disease. Mycoplasma hyopneumoniae colonizes the swine epithelium and causes a pneumonia that predisposes the host to enhanced disease from other pathogens. How M. hyopneumoniae responds to changing environments in the respiratory tract during disease progression is not known. In fact, little is known concerning the capabilities of mycoplasmas to respond to changing growth environments. With limited genes, mycoplasmas are thought to possess only a few mechanisms for gene regulation. A microarray consisting of 632 of the 698 open reading frames of M. hyopneumoniae was constructed and used to study gene expression differences during a temperature shift from 37°C to 42°C, a temperature swing that might be encountered during disease. To enhance sensitivity, a unique hexamer primer set was employed for generating cDNA from only mRNA species. Our analysis identified 91 genes that had significant transcriptional differences in response to heat shock conditions (P < 0.01) with an estimated false-discovery rate of 4 percent. Thirty-three genes had a change threshold of 1.5-fold or greater. Many of the heat shock proteins previously characterized in other bacteria were identified as significant in this study as well. A proportion of the identified genes (54 of 91) currently have no assigned function. PMID:16368969

  12. Genital Mycoplasma infection among Mexican women with systemic lupus erythematosus.

    PubMed

    Méndez-Martínez, Socorro; García-Carrasco, Mario; Cedillo-Ramírez, María L; Mendoza-Pinto, Claudia; Etchegaray-Morales, Ivet; Gil-Juárez, Constantino; Montiel-Jarquín, Álvaro J; Taboada-Cole, Alejandro; Jiménez-Herrera, Erick A; Muñóz-Guarneros, Margarita; Cervera, Ricard

    2017-07-01

    To assess the prevalence of genital Mycoplasma spp. among women with systemic lupus erythematosus (SLE) and to identify factors associated with such infection. A cross-sectional study was conducted among patients with SLE and healthy women who attended a hospital in Puebla, Mexico, between July 29, 2014, and January 4, 2015. All participants were aged 18 years or older and sexually active. A structured interview assessed sociodemographic, obstetric, gynecologic, and clinical characteristics. Disease activity was evaluated using the Mexican SLE Disease Activity Index. Polymerase chain reaction was used to detect the presence of Mycoplasma spp. in genital samples. Ureaplasma urealyticum was the only genital mycoplasma detected; it was present in 32 (24.6%) of 130 patients with SLE and 12 (12.8%) of 94 healthy women. Patients with SLE had increased odds of infection (odds ratio 2.120, 95% confidence interval 1.046-4.296). Among patients with SLE, multiparity was more common in those with U. urealyticum infection (P=0.043). One-quarter of women with SLE had genital infection with U. urealyticum. An association was found between infection and multiparity among women with SLE. © 2017 International Federation of Gynecology and Obstetrics.

  13. Isolation and immunological detection of Mycoplasma ovipneumoniae in sheep with atypical pneumonia, and lack of a role for Mycoplasma arginini.

    PubMed

    Lin, Y-C; Miles, R J; Nicholas, R A J; Kelly, D P; Wood, A P

    2008-06-01

    Mycoplasma ovipneumoniae NCTC 10151(T) and four new isolates from UK sheep flocks were compared. Only glucose and pyruvate were used as energy sources by the five strains: glucose was the best energy source for the type strain, pyruvate supported better growth of the new strains. Whole cell protein patterns and antigenic profiles showed high similarity between all five strains. The new isolates fell into two groups in ELISA tests. Serum samples from 30 pneumonic sheep were assessed for M. ovipneumoniae infection and Mycoplasma arginini co-infection. Fourteen (out of 30) serum samples were positive for M. ovipneumoniae both by ELISA and immunoblotting. Twelve antigenic proteins of M. ovipneumoniae were detected in infected serum samples: the antigen patterns were unique, with between one and at least seven occurring in any one sample. All serum samples were designated as negative for M. arginini antibodies by both ELISA and immunoblotting.

  14. Detection, Characterization, and Molecular Typing of Human Mycoplasma spp. from Major Hospitals in Cairo, Egypt

    PubMed Central

    Metwally, Mirihan A.; Yassin, Aymen S.; Essam, Tamer M.; Hamouda, Hayam M.; Amin, Magdy A.

    2014-01-01

    Mycoplasmas are fastidious slow growing organisms lacking a cell wall and mostly isolated from the mucosal surfaces of the respiratory and genitourinary tracts. There is a dearth of information regarding clinical Mycoplasma spp. isolates among Egyptian patients. A total of 170 samples were collected from patients and apparently healthy personnel in local public hospitals in Cairo, Egypt. Isolation of Mycoplasma spp. was carried out using appropriate culture media and further identification was carried out by biochemical tests followed by serotyping using specific antisera. Confirmation was done by PCR for detection of different Mycoplasma spp. using genus-specific primers targeting 16S ribosomal RNA gene. Characterization of the antibiotic resistance and sensitivity pattern against different antimicrobials was carried out using disc diffusion test. The results indicated the presence of six Mycoplasma spp. in 22.94% of the samples. Mycoplasmas were detected more frequently in throat swabs than sputum. Mycoplasma pneumoniae was highly sensitive to macrolides and quinolones but less sensitive to aminoglycosides and tetracyclines. Molecular techniques were found to be of more rapid, highly sensitive, able to detect nonviable organisms, and cost effective. These results shed light on difficulties of Mycoplasma detection and the superiority of molecular techniques over culture. PMID:25506614

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

    PubMed

    Jimena, Otero Negrete; Laura, Jaramillo Meza; Elena, Miranda Morales Rosa; Alonso, Navarro Hernández Jaime; Teresa, Quintero Martínez María

    2009-11-01

    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.

  16. Seroprevalence of Mycoplasma sp. in farmed bison (Bison bison) herds in western Canada

    USDA-ARS?s Scientific Manuscript database

    Mycoplasma bovis is emerging as an important pathogen of farmed bison in North America, associated with high morbidity and mortality. An in-house enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies against Mycoplasma sp. in bison sera. The aims of the study were to estimate ...

  17. Severe anemia associated with Mycoplasma wenyonii infection in a mature cow

    PubMed Central

    Genova, Suzanne G.; Streeter, Robert N.; Velguth, Karen E.; Snider, Timothy A.; Kocan, Katherine M.; Simpson, Katharine M.

    2011-01-01

    The clinical findings, diagnostic tests, and treatment of clinical anemia in a mature Angus cow infected with the hemoplasma Mycoplasma wenyonii are described. Mycoplasma wenyonii has been previously reported to cause clinical anemia in young or splenectomized cattle; however, infection has not been associated with severe anemia in mature animals. PMID:22379205

  18. World Health Organization International Standard To Harmonize Assays for Detection of Mycoplasma DNA

    PubMed Central

    Baylis, Sally A.; Hanschmann, Kay-Martin; Montag-Lessing, Thomas; Chudy, Michael; Kreß, Julia; Ulrych, Ursula; Czurda, Stefan; Rosengarten, Renate

    2015-01-01

    Nucleic acid amplification technique (NAT)-based assays (referred to here as NAT assays) are increasingly used as an alternative to culture-based approaches for the detection of mycoplasma contamination of cell cultures. Assay features, like the limit of detection or quantification, vary widely between different mycoplasma NAT assays. Biological reference materials may be useful for harmonization of mycoplasma NAT assays. An international feasibility study included lyophilized preparations of four distantly related mycoplasma species (Acholeplasma laidlawii, Mycoplasma fermentans, M. orale, M. pneumoniae) at different concentrations which were analyzed by 21 laboratories using 26 NAT assays with a qualitative, semiquantitative, or quantitative design. An M. fermentans preparation was shown to decrease the interassay variation when used as a common reference material. The preparation was remanufactured and characterized in a comparability study, and its potency (in NAT-detectable units) across different NATs was determined. The World Health Organization (WHO) Expert Committee on Biological Standardization (ECBS) established this preparation to be the “1st World Health Organization international standard for mycoplasma DNA for nucleic acid amplification technique-based assays designed for generic mycoplasma detection” (WHO Tech Rep Ser 987:42, 2014) with a potency of 200,000 IU/ml. This WHO international standard is now available as a reference preparation for characterization of NAT assays, e.g., for determination of analytic sensitivity, for calibration of quantitative assays in a common unitage, and for defining regulatory requirements in the field of mycoplasma testing. PMID:26070671

  19. Detection, characterization, and molecular typing of human Mycoplasma spp. from major hospitals in Cairo, Egypt.

    PubMed

    Metwally, Mirihan A; Yassin, Aymen S; Essam, Tamer M; Hamouda, Hayam M; Amin, Magdy A

    2014-01-01

    Mycoplasmas are fastidious slow growing organisms lacking a cell wall and mostly isolated from the mucosal surfaces of the respiratory and genitourinary tracts. There is a dearth of information regarding clinical Mycoplasma spp. isolates among Egyptian patients. A total of 170 samples were collected from patients and apparently healthy personnel in local public hospitals in Cairo, Egypt. Isolation of Mycoplasma spp. was carried out using appropriate culture media and further identification was carried out by biochemical tests followed by serotyping using specific antisera. Confirmation was done by PCR for detection of different Mycoplasma spp. using genus-specific primers targeting 16S ribosomal RNA gene. Characterization of the antibiotic resistance and sensitivity pattern against different antimicrobials was carried out using disc diffusion test. The results indicated the presence of six Mycoplasma spp. in 22.94% of the samples. Mycoplasmas were detected more frequently in throat swabs than sputum. Mycoplasma pneumoniae was highly sensitive to macrolides and quinolones but less sensitive to aminoglycosides and tetracyclines. Molecular techniques were found to be of more rapid, highly sensitive, able to detect nonviable organisms, and cost effective. These results shed light on difficulties of Mycoplasma detection and the superiority of molecular techniques over culture.

  20. Experimental infection of BHK21 and Vero cell lines with different Mycoplasma spp.

    PubMed

    Netto, Cristiane; Soccol, Vanete Thomaz; Sepulveda, Lya Madureira; Timenetsky, Jorge

    2014-01-01

    Mycoplasma spp, belongs to the class Mollicutes and is capable to produce alterations in cellular cultures causing damages to the biotechnological industry. Bioproducts generally require two essential inputs, bovine serum and cells. The study herein aims to evaluate the mycoplasma concentrations that affect the growing of BHK21 and Vero cells. The species used were: Mycoplasma orale, M. salivarium, M. arginini and M. hyorhinis, cultivated in a SP4 media. Two contamination tests were performed with BHK21 and Vero cells and one of them applied different concentrations of mycoplasma. In the first one, mycoplasma was applied at the day zero and, in the second one, the contamination was performed after the monolayer establishment. The both cellular cultures presented cytopathic effects with mycoplasma contamination, but the Vero cells suffered more damages than the BHK21 ones. It was also observed that the severity of the cytopathic effect depended on the mycoplasma specie, on the concentration and on the time of contact with the cellular culture, which evidences the importance of controlling the presence of mycoplasma in biotechnological industries.

  1. World Health Organization International Standard To Harmonize Assays for Detection of Mycoplasma DNA.

    PubMed

    Nübling, C Micha; Baylis, Sally A; Hanschmann, Kay-Martin; Montag-Lessing, Thomas; Chudy, Michael; Kreß, Julia; Ulrych, Ursula; Czurda, Stefan; Rosengarten, Renate

    2015-09-01

    Nucleic acid amplification technique (NAT)-based assays (referred to here as NAT assays) are increasingly used as an alternative to culture-based approaches for the detection of mycoplasma contamination of cell cultures. Assay features, like the limit of detection or quantification, vary widely between different mycoplasma NAT assays. Biological reference materials may be useful for harmonization of mycoplasma NAT assays. An international feasibility study included lyophilized preparations of four distantly related mycoplasma species (Acholeplasma laidlawii, Mycoplasma fermentans, M. orale, M. pneumoniae) at different concentrations which were analyzed by 21 laboratories using 26 NAT assays with a qualitative, semiquantitative, or quantitative design. An M. fermentans preparation was shown to decrease the interassay variation when used as a common reference material. The preparation was remanufactured and characterized in a comparability study, and its potency (in NAT-detectable units) across different NATs was determined. The World Health Organization (WHO) Expert Committee on Biological Standardization (ECBS) established this preparation to be the "1st World Health Organization international standard for mycoplasma DNA for nucleic acid amplification technique-based assays designed for generic mycoplasma detection" (WHO Tech Rep Ser 987:42, 2014) with a potency of 200,000 IU/ml. This WHO international standard is now available as a reference preparation for characterization of NAT assays, e.g., for determination of analytic sensitivity, for calibration of quantitative assays in a common unitage, and for defining regulatory requirements in the field of mycoplasma testing.

  2. Characterization studies on mycoplasmas isolated from bovine mastitis and the bovine respiratory tract.

    PubMed

    Dellinger, J D; Jasper, D E; Ilić, M

    1977-07-01

    Mycoplasmas isolated from bovine mastitis in California were classified into five distinct species. These included Mycoplasma bovis, M. bovigenitalium, M. alkalescens, M. canadenfe, and an unidentified strain, ST-6. Strains frequently recovered from the nose of young calves proved to be M. arginini, M. bovirhinis was recovered from the respiratory tract but was not a common finding.

  3. Mycoplasma membrane lipoproteins induced proinflammatory cytokines by a mechanism distinct from that of lipopolysaccharide.

    PubMed Central

    Rawadi, G; Roman-Roman, S

    1996-01-01

    To gain a clear understanding of the mechanisms by which mycoplasmas induced the expression of proinflammatory cytokines in monocytic cells, we have studied the induction of interleukin-1 beta (IL-1 beta), tumor necrosis factor alpha, and IL-6 by mycoplasmas in three distinct human myelomonocytic cell lines in comparison with induction by lipopolysaccharide (LPS). HL-60 cell line did not release cytokines when induced with either LPS or mycoplasmas. In contrast to LPS, mycoplasmas failed to increase the weak levels of tumor necrosis factor alpha secreted by phorbol myristate acetate-differentiated U937 cells. In addition, Northern (RNA) blot analysis of cytokine expression in these cells showed that the induction of IL-1 beta by mycoplasmas involves, unlike that by LPS, posttranscriptional events. Interestingly, in THP-1 cells, cytokine induction pathways triggered by mycoplasmas remained operational under conditions where LPS pathways were abolished, suggesting functional independence. The study of cytokine-inducing activity displayed by distinct fractions derived from a series of different mycoplasma species demonstrated that lipid membrane constituents were largely responsible for these effects. Finally, we have demonstrated that tyrosine phosphorylation is a crucial event in the mycoplasma-mediated induction of proinflammatory cytokines in either THP-1 cells or human monocytes. PMID:8550219

  4. Experimental infection of BHK21 and Vero cell lines with different Mycoplasma spp

    PubMed Central

    Netto, Cristiane; Soccol, Vanete Thomaz; Sepulveda, Lya Madureira; Timenetsky, Jorge

    2014-01-01

    Mycoplasma spp, belongs to the class Mollicutes and is capable to produce alterations in cellular cultures causing damages to the biotechnological industry. Bioproducts generally require two essential inputs, bovine serum and cells. The study herein aims to evaluate the mycoplasma concentrations that affect the growing of BHK21 and Vero cells. The species used were: Mycoplasma orale, M. salivarium, M. arginini and M. hyorhinis, cultivated in a SP4 media. Two contamination tests were performed with BHK21 and Vero cells and one of them applied different concentrations of mycoplasma. In the first one, mycoplasma was applied at the day zero and, in the second one, the contamination was performed after the monolayer establishment. The both cellular cultures presented cytopathic effects with mycoplasma contamination, but the Vero cells suffered more damages than the BHK21 ones. It was also observed that the severity of the cytopathic effect depended on the mycoplasma specie, on the concentration and on the time of contact with the cellular culture, which evidences the importance of controlling the presence of mycoplasma in biotechnological industries. PMID:25763061

  5. Amyloid-beta peptide degradation in cell cultures by mycoplasma contaminants

    PubMed Central

    Zhao, Haitian; Dreses-Werringloer, Ute; Davies, Peter; Marambaud, Philippe

    2008-01-01

    Background Cell cultures have become an indispensable tool in Alzheimer's disease research for studying amyloid-β (Aβ) metabolism. It is estimated that up to 35% of cell cultures in current use are infected with various mycoplasma species. In contrast with common bacterial and fungal infections, contaminations of cell cultures with mycoplasmas represent a challenging issue in terms of detectability and prevention. Mycoplasmas are the smallest and simplest self-replicating bacteria and the consequences of an infection for the host cells are variable, ranging from no apparent effect to induction of apoptosis. Findings Here we present evidence that mycoplasmas from a cell culture contamination are able to efficiently and rapidly degrade extracellular Aβ. As a result, we observed no accumulation of Aβ in the conditioned medium of mycoplasma-positive cells stably transfected with the amyloid-β precursor protein (APP). Importantly, eradication of the mycoplasma contaminant – identified as M. hyorhinis – by treatments with a quinolone-based antibiotic, restored extracellular Aβ accumulation in the APP-transfected cells. Conclusion These data show that mycoplasmas degrade Aβ and thus may represent a significant source of variability when comparing extracellular Aβ levels in different cell lines. On the basis of these results, we recommend assessment of mycoplasma contaminations prior to extracellular Aβ level measurements in cultured cells. PMID:18710491

  6. Amyloid-beta peptide degradation in cell cultures by mycoplasma contaminants.

    PubMed

    Zhao, Haitian; Dreses-Werringloer, Ute; Davies, Peter; Marambaud, Philippe

    2008-06-30

    Cell cultures have become an indispensable tool in Alzheimer's disease research for studying amyloid-beta (Abeta) metabolism. It is estimated that up to 35% of cell cultures in current use are infected with various mycoplasma species. In contrast with common bacterial and fungal infections, contaminations of cell cultures with mycoplasmas represent a challenging issue in terms of detectability and prevention. Mycoplasmas are the smallest and simplest self-replicating bacteria and the consequences of an infection for the host cells are variable, ranging from no apparent effect to induction of apoptosis. Here we present evidence that mycoplasmas from a cell culture contamination are able to efficiently and rapidly degrade extracellular Abeta. As a result, we observed no accumulation of Abeta in the conditioned medium of mycoplasma-positive cells stably transfected with the amyloid-beta precursor protein (APP). Importantly, eradication of the mycoplasma contaminant - identified as M. hyorhinis - by treatments with a quinolone-based antibiotic, restored extracellular Abeta accumulation in the APP-transfected cells. These data show that mycoplasmas degrade Abeta and thus may represent a significant source of variability when comparing extracellular Abeta levels in different cell lines. On the basis of these results, we recommend assessment of mycoplasma contaminations prior to extracellular Abeta level measurements in cultured cells.

  7. Host Cell Responses to Persistent Mycoplasmas - Different Stages in Infection of HeLa Cells with Mycoplasma hominis

    PubMed Central

    Hopfe, Miriam; Deenen, René; Degrandi, Daniel; Köhrer, Karl; Henrich, Birgit

    2013-01-01

    Mycoplasma hominis is a facultative human pathogen primarily associated with bacterial vaginosis and pelvic inflammatory disease, but it is also able to spread to other sites, leading to arthritis or, in neonates, meningitis. With a minimal set of 537 annotated genes, M. hominis is the second smallest self-replicating mycoplasma and thus an ideal model organism for studying the effects of an infectious agent on its host more closely. M. hominis adherence, colonisation and invasion of HeLa cells were characterised in a time-course study using scanning electron microscopy, confocal microscopy and microarray-based analysis of the HeLa cell transcriptome. At 4 h post infection, cytoadherence of M. hominis to the HeLa cell surface was accompanied by differential regulation of 723 host genes (>2 fold change in expression). Genes associated with immune responses and signal transduction pathways were mainly affected and components involved in cell-cycle regulation, growth and death were highly upregulated. At 48 h post infection, when mycoplasma invasion started, 1588 host genes were differentially expressed and expression of genes for lysosome-specific proteins associated with bacterial lysis was detected. In a chronically infected HeLa cell line (2 weeks), the proportion of intracellular mycoplasmas reached a maximum of 10% and M. hominis-filled protrusions of the host cell membrane were seen by confocal microscopy, suggesting exocytotic dissemination. Of the 1972 regulated host genes, components of the ECM-receptor interaction pathway and phagosome-related integrins were markedly increased. The immune response was quite different to that at the beginning of infection, with a prominent induction of IL1B gene expression, affecting pathways of MAPK signalling, and genes connected with cytokine-cytokine interactions and apoptosis. These data show for the first time the complex, time-dependent reaction of the host directed at mycoplasmal clearance and the counter measures of

  8. Host cell responses to persistent mycoplasmas--different stages in infection of HeLa cells with Mycoplasma hominis.

    PubMed

    Hopfe, Miriam; Deenen, René; Degrandi, Daniel; Köhrer, Karl; Henrich, Birgit

    2013-01-01

    Mycoplasma hominis is a facultative human pathogen primarily associated with bacterial vaginosis and pelvic inflammatory disease, but it is also able to spread to other sites, leading to arthritis or, in neonates, meningitis. With a minimal set of 537 annotated genes, M. hominis is the second smallest self-replicating mycoplasma and thus an ideal model organism for studying the effects of an infectious agent on its host more closely. M. hominis adherence, colonisation and invasion of HeLa cells were characterised in a time-course study using scanning electron microscopy, confocal microscopy and microarray-based analysis of the HeLa cell transcriptome. At 4 h post infection, cytoadherence of M. hominis to the HeLa cell surface was accompanied by differential regulation of 723 host genes (>2 fold change in expression). Genes associated with immune responses and signal transduction pathways were mainly affected and components involved in cell-cycle regulation, growth and death were highly upregulated. At 48 h post infection, when mycoplasma invasion started, 1588 host genes were differentially expressed and expression of genes for lysosome-specific proteins associated with bacterial lysis was detected. In a chronically infected HeLa cell line (2 weeks), the proportion of intracellular mycoplasmas reached a maximum of 10% and M. hominis-filled protrusions of the host cell membrane were seen by confocal microscopy, suggesting exocytotic dissemination. Of the 1972 regulated host genes, components of the ECM-receptor interaction pathway and phagosome-related integrins were markedly increased. The immune response was quite different to that at the beginning of infection, with a prominent induction of IL1B gene expression, affecting pathways of MAPK signalling, and genes connected with cytokine-cytokine interactions and apoptosis. These data show for the first time the complex, time-dependent reaction of the host directed at mycoplasmal clearance and the counter measures of

  9. Occurrence of mycoplasmas in free-ranging birds of prey in Germany.

    PubMed

    Lierz, M; Hagen, N; Hernadez-Divers, S J; Hafez, H M

    2008-10-01

    Mycoplasmas are well-known avian pathogens of poultry and some passerines. Although reported in birds of prey, their role as pathogens is still unclear. Healthy, free-ranging raptor nestlings sampled during a routine ringing (banding) program, and birds of prey from rehabilitation centers, tested positive for Mycoplasma spp. by culture and a genus-specific polymerase chain reaction (PCR). Given the lack of clinical signs and disease, we suggest that mycoplasmas in raptors may be commensal rather than pathogenic. Using immunobinding assay and species-specific PCR tests, Mycoplasma buteonis, M. falconis, and M. gypis were identified; M. falconis was only detected in falcons. Additionally, some isolates could not be identified. This is the first report of Mycoplasma spp. isolations from Western Marsh Harriers (Circus aeroginosus), a Eurasian Hobby (Falco subbuteo), and a Barn Owl (Tyto alba).

  10. Identification and characterization of novel Mycoplasma spp. belonging to the hominis group from griffon vultures.

    PubMed

    Lecis, R; Chessa, B; Cacciotto, C; Addis, M F; Coradduzza, E; Berlinguer, F; Muzzeddu, M; Lierz, M; Carcangiu, L; Pittau, M; Alberti, A

    2010-08-01

    Mycoplasmas are commensals and pathogens of various avian species, and are also regularly found in birds of prey, although their significance to birds' health remains unclear. Here we describe two novel Mycoplasma isolated from the upper respiratory tract of four Eurasian griffon vultures (Gyps fulvus) housed in a wildlife recovery centre in Sardinia (Italy). By sequencing the 16S rRNA gene and the entire 16S/23S intergenic spacer region, the new strains were classified within the Mycoplasma taxonomy at the group and cluster levels, showing that the two isolates fall into the Mycoplasma synoviae and Mycoplasma hominis clusters of the hominis group, respectively. We combined molecular tools and immunoblotting methods in order to further characterize these isolates, and antigenic analyses overall confirmed the molecular findings. Different levels of pathogenicity and prevalence of these strains might have different implications for the conservation and reintroduction of vultures. Copyright 2010 Elsevier Ltd. All rights reserved.

  11. MOLECULAR IDENTIFICATION AND SEQUENCE CHARACTERIZATION OF MYCOPLASMAS IN FREE-LIVING BIRDS OF PREY.

    PubMed

    Lecis, Roberta; Secci, Fabio; Mandas, Lucio; Muzzeddu, Marco; Pittau, Marco; Alberti, Alberti

    2016-09-01

    Mycoplasma spp. have been detected in birds of prey, but their prevalence in free living raptors and their significance to birds' health need further investigation. Molecular techniques have been increasingly used to identify mycoplasmas in various avian species, due to the fastidious nature of these pathogens hampering traditional bacteriologic tests. This study reports the identification of 23 novel mycoplasma sequences during the monitoring of 62 birds of prey on admission to wildlife centers in Sardinia, Italy. Molecular investigation performed on pharyngeal swabs revealed 26 birds positive to Mycoplasma (42%). Sequence analysis based on 16S rRNA, 16S-23S rRNA intergenic spacer, and RNA polymerase β subunit (rpoB) gene highlighted cluster assignment and phylogenetic relationships among the identified types, classified within the hominis group. Additionally, Ornithobacterium rhinotracheale , associated with respiratory disease in poultry, was identified in 17 birds (27%). Potential coinfection and mycoplasma opportunistic nature present implications for raptor species conservation.

  12. Lesions associated with a novel Mycoplasma sp. in California sea lions (Zalophus californianus) undergoing rehabilitation.

    PubMed

    Haulena, Martin; Gulland, Frances M D; Lawrence, Judith A; Fauquier, Deborah A; Jang, Spencer; Aldridge, Brian; Spraker, Terry; Thomas, Linda C; Brown, Daniel R; Wendland, Lori; Davidson, Maureen K

    2006-01-01

    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.

  13. Mycoplasma species isolated from harbor porpoises (Phocoena phocoena) and a Sowerby's beaked whale (Mesoplodon bidens) stranded in Scottish waters.

    PubMed

    Foster, Geoffrey; McAuliffe, Laura; Dagleish, Mark P; Barley, Jason; Howie, Fiona; Nicholas, Robin A J; Ayling, Roger D

    2011-01-01

    Mycoplasma species were recovered from 10 cetacean carcasses that stranded around Scotland. Mycoplasma phocicerebrale was isolated from the lungs of three harbor porpoises (Phocoena phocoena) as well as from the liver of one of these animals. Novel Mycoplasma spp. were isolated from the lungs of five additional harbor porpoises and the kidney of another. In addition an isolate closely related to Mycoplasma species 13CL was obtained from the kidney of a Sowerby's beaked whale (Mesoplodon bidens). The role of these Mycoplasma species in the disease of cetaceans, their host specificity, diversity, and any relation to cetacean strandings are unknown.

  14. Antibody responses of swine following infection with Mycoplasma hyopneumoniae, M. hyorhinis, M. hyosynoviae and M. flocculare.

    PubMed

    Gomes Neto, João Carlos; Strait, Erin L; Raymond, Matthew; Ramirez, Alejandro; Minion, F Chris

    2014-11-07

    Several mycoplasma species possessing a range of virulence have been described in swine. The most commonly described are Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, Mycoplasma hyosynoviae, and Mycoplasma flocculare. They are ubiquitious in many pig producing areas of the world, and except for M. hyopneumoniae, commercial antibody-based assays are lacking for most of these. Antibody cross-reactivity among these four mycoplasma species is not well characterized. Recently, the use of pen-based oral fluids for herd surveillance is of increasing interest. Thus, this study sought to measure pig antibody responses and the level of cross-reactivity in serum and pen-based oral fluids after challenge with four species of swine mycoplasmas. Four groups of four mycoplasma-free growing pigs were separately inoculated with the different mycoplasma species. Pen-based oral fluids and serum samples were collected weekly until necropsy. Species-specific Tween 20 ELISAs were used to measure antibody responses along with four other commercial M. hyopneumoniae ELISAs. Animals from all groups seroconverted to the challenge species of mycoplasma and no evidence of cross-contamination was observed. A delayed antibody response was seen with all but M. hyorhinis-infected pigs. Cross-reactive IgG responses were detected in M. hyopneumoniae- and M. flocculare-infected animals by the M. hyorhinis Tween 20 ELISA, while sera from M. hyosynoviae and M. flocculare-infected pigs were positive in one commercial assay. In pen-based oral fluids, specific anti-M. hyopneumoniae IgA responses were detected earlier after infection than serum IgG responses. In summary, while some antibody-based assays may have the potential for false positives, evidence of this was observed in the current study.

  15. Molecular characterisation of Mycoplasma species isolated from the genital tract of Dorper sheep in South Africa.

    PubMed

    Kalshingi, Habu A; Bosman, Anna-Mari; Gouws, Johan; van Vuuren, Moritz

    2015-06-08

    Biochemical and molecular analysis were conducted on 34 strains of Mycoplasma species isolated between 2003 and 2009 from the genital tract of clinically healthy Dorper sheep and sheep with ulcerative vulvitis and balanitis. Earlier publications identified the causative agent as Mycoplasma mycoides mycoides large colony (MmmLC) and Arcanobacterium pyogenes. The aims of the study were to characterise Mycoplasma species isolated from the genital tract of Dorper sheep with polymerase chain reaction assay, cloning and gene sequencing. Basic Local Alignment Search Tool (BLAST) results revealed six predominant Mycoplasma species: Mycoplasma arginini, Mycoplasma bovigenitalium, Arcanobacterium laidlawii, MmmLC, Mycoplasma sp. ovine/caprine serogroup II and M. canadense. Sequencing of the 34 isolates were analysed using phylogenetic methods, and 18 (50%) were identified as M. arginini with 99% - 100% similarity to M. arginini from England and Sweden. Six isolates showed 99% similarity to M. bovigenitalium strains from Turkey and Germany. Two isolates had 99% similarity to an M. sp. ovine/caprine sero group II from the United Kingdom. BLAST for two isolates revealed 99% similarity to Acholeplasma laidlawii from India, another two were 99% similar to MmmLC strain from Sweden, two showed 98% similarity to Mycoplasma sp. Usp 120 from Brazil, and two isolates have a 97% - 99% similarity to M. mm. Jcv1 strain from the United States of America. Finally, one isolate showed similarity of 99% to Mycoplasma canadense strain from Italy. The findings support the hypothesis that ulcerative vulvitis and balanitis of Dorper sheep in South Africa (SA) is a multifactorial disease with involvement of different Mycoplasma species.

  16. Mycoplasma hominis and Mycoplasma genitalium in the Vaginal Microbiota and Persistent High-Risk Human Papillomavirus Infection.

    PubMed

    Adebamowo, Sally N; Ma, Bing; Zella, Davide; Famooto, Ayotunde; Ravel, Jacques; Adebamowo, Clement

    2017-01-01

    Recent studies have suggested that the vaginal microenvironment plays a role in persistence of high-risk human papillomavirus (hrHPV) infection and thus cervical carcinogenesis. Furthermore, it has been shown that some mycoplasmas are efficient methylators and may facilitate carcinogenesis through methylation of hrHPV and cervical somatic cells. We examined associations between prevalence and persistence of Mycoplasma spp. in the vaginal microbiota, and prevalent as well as persistent hrHPV infections. We examined 194 Nigerian women who were tested for hrHPV infection using SPF25/LiPA10 and we identified Mycoplasma genitalium and Mycoplasma hominis in their vaginal microbiota established by sequencing the V3-V4 hypervariable regions of the 16S rRNA gene. We defined the prevalence of M. genitalium, M. hominis, and hrHPV based on positive result of baseline tests, while persistence was defined as positive results from two consecutive tests. We used exact logistic regression models to estimate associations between Mycoplasma spp. and hrHPV infections. The mean (SD) age of the study participants was 38 (8) years, 71% were HIV positive, 30% M. genitalium positive, 45% M. hominis positive, and 40% hrHPV positive at baseline. At follow-up, 16% of the women remained positive for M. genitalium, 30% for M. hominis, and 31% for hrHPV. There was a significant association between persistent M. hominis and persistent hrHPV (OR 8.78, 95% CI 1.49-51.6, p 0.01). Women who were positive for HIV and had persistent M. hominis had threefold increase in the odds of having persistent hrHPV infection (OR 3.28, 95% CI 1.31-8.74, p 0.008), compared to women who were negative for both. We found significant association between persistent M. hominis in the vaginal microbiota and persistent hrHPV in this study, but we could not rule out reverse causation. Our findings need to be replicated in larger, longitudinal studies and if confirmed, could have important diagnostic and therapeutic

  17. Mycoplasma-dependent activation of normal mouse lymphocytes: requirement for functional T lymphocytes in the cytotoxicity reaction mediated by Mycoplasma arthritidis.

    PubMed Central

    Cole, B C; Aldridge, K E; Sullivan, G J; Ward, J R

    1980-01-01

    Syngeneic and allogeneic target cells were killed in the presence of CBA mouse lymphocytes and viable Mycoplasma arthritidis. Medium supplementation had no effect on the response. Nonviable M. arthritidis was also capable of stimulating lymphocytotoxicity, although to a much lesser extent. Cytotoxicity was shown to be largely dependent upon the lymphocytes, since lymphocytes preincubated with mycoplasmas and treated to remove remaining organisms were highly toxic to target cells, whereas supernatants prepared from lymphocyte/mycoplasma mixtures exhibited minimal effects. A 6-h exposure of lymphocytes to mycoplasmas at a ratio of 100:1 was sufficient for commitment to target cell killing. Functional lymphocytes were required for the reaction, since gamma-irradiated lymphocytes did not develop cytotoxic potential despite the fact that the mycoplasmas replicated equally well in the presence of these and untreated lymphocytes. Furthermore, lymphocytes already activated with mycoplasmas lost cytotoxic potential after disruption. The kinetics and degree of lymphocytotoxicity induced by M. arthritidis and phytohemagglutinin toward 51Cr-labeled syngeneic fibroblasts were similar. Removal of most B cells and other adherent cells by column separation did not abrogate the cytotoxic effect. Lymphocyte suspensions treated with anti-Thy 1 antiserum and complement exhibited a marked decrease in their cytotoxic potential when added to labeled target cells in the presence of M. arthritidis. We conclude that the cytotoxic reaction is dependent upon the T-lymphocyte subpopulation. PMID:6969227

  18. Genomic characterization of symbiotic mycoplasmas from the stomach of deep-sea isopod bathynomus sp.

    PubMed

    Wang, Yong; Huang, Jiao-Mei; Wang, Shao-Lu; Gao, Zhao-Ming; Zhang, Ai-Qun; Danchin, Antoine; He, Li-Sheng

    2016-09-01

    Deep-sea isopod scavengers such as Bathynomus sp. are able to live in nutrient-poor environments, which is likely attributable to the presence of symbiotic microbes in their stomach. In this study we recovered two draft genomes of mycoplasmas, Bg1 and Bg2, from the metagenomes of the stomach contents and stomach sac of a Bathynomus sp. sample from the South China Sea (depth of 898 m). Phylogenetic trees revealed a considerable genetic distance to other mycoplasma species for Bg1 and Bg2. Compared with terrestrial symbiotic mycoplasmas, the Bg1 and Bg2 genomes were enriched with genes encoding phosphoenolpyruvate-dependent phosphotransferase systems (PTSs) and sodium-driven symporters responsible for the uptake of sugars, amino acids and other carbohydrates. The genome of mycoplasma Bg1 contained sialic acid lyase and transporter genes, potentially enabling the bacteria to attach to the stomach sac and obtain organic carbons from various cell walls. Both of the mycoplasma genomes contained multiple copies of genes related to proteolysis and oligosaccharide degradation, which may help the host survive in low-nutrient conditions. The discovery of the different types of mycoplasma bacteria in the stomach of this deep-sea isopod affords insights into symbiotic model of deep-sea animals and genomic plasticity of mycoplasma bacteria. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  19. The effects of mycoplasma contamination upon the ability to form bioengineered 3D kidney cysts.

    PubMed

    DesRochers, Teresa M; Kuo, Ivana Y; Kimmerling, Erica P; Ehrlich, Barbara E; Kaplan, David L

    2015-01-01

    Mycoplasma contamination of cell cultures is a pervasive, often undiagnosed and ignored problem in many laboratories that can result in reduced cell proliferation and changes in gene expression. Unless contamination is specifically suspected, it is often undetected in two dimensional (2D) cultures and the resulting effects of mycoplasma contamination are rarely appreciated and can lead to incorrect conclusions. Three dimensional (3D) tissue cultures are increasingly utilized to explore tissue development and phenotype. However, 3D cultures are more complex than 2D cell cultures and require a more controlled cellular environment in order to generate structures necessary to mimic in vivo responses and are often maintained for longer time periods. Changes to the microenvironment are assumed to have a more extreme effect upon the success of 3D tissue cultures than 2D cell cultures, but the effects of mycoplasma have not been studied. To test this hypothesis, we grew 2D cell cultures and 3D tissues from pig kidney epithelial cells (LLC-PK1) that were contaminated with mycoplasma and the same stock of cells after mycoplasma removal. We did not observe an effect of mycoplasma contamination on proliferation in 2D monolayer cell culture. However, cyst formation in 3D tissues was altered, with effects upon the number, size and structure of cysts formed. These data serve to reinforce the necessity of testing cell stocks for mycoplasma contamination.

  20. Mycoplasma corogypsi-associated polyarthritis and tenosynovitis in black vultures (Coragyps atratus).

    PubMed

    Van Wettere, A J; Ley, D H; Scott, D E; Buckanoff, H D; Degernes, L A

    2013-03-01

    Three wild American black vultures (Coragyps atratus) were presented to rehabilitation centers with swelling of multiple joints, including elbows, stifles, hocks, and carpal joints, and of the gastrocnemius tendons. Cytological examination of the joint fluid exudate indicated heterophilic arthritis. Radiographic examination in 2 vultures demonstrated periarticular soft tissue swelling in both birds and irregular articular surfaces with subchondral bone erosion in both elbows in 1 bird. Prolonged antibiotic therapy administered in 2 birds did not improve the clinical signs. Necropsy and histological examination demonstrated a chronic lymphoplasmacytic arthritis involving multiple joints and gastrocnemius tenosynovitis. Articular lesions varied in severity and ranged from moderate synovitis and cartilage erosion and fibrillation to severe synovitis, diffuse cartilage ulceration, subchondral bone loss and/or sclerosis, pannus, synovial cysts, and epiphyseal osteomyelitis. No walled bacteria were observed or isolated from the joints. However, mycoplasmas polymerase chain reactions were positive in at least 1 affected joint from each bird. Mycoplasmas were isolated from joints of 1 vulture that did not receive antibiotic therapy. Sequencing of 16S rRNA gene amplicons from joint samples and the mycoplasma isolate identified Mycoplasma corogypsi in 2 vultures and was suggestive in the third vulture. Mycoplasma corogypsi identification was confirmed by sequencing the 16S-23S intergenic spacer region of mycoplasma isolates. This report provides further evidence that M. corogypsi is a likely cause of arthritis and tenosynovitis in American black vultures. Cases of arthritis and tenosynovitis in New World vultures should be investigated for presence of Mycoplasma spp, especially M. corogypsi.

  1. Extracellular membrane vesicles secreted by mycoplasma Acholeplasma laidlawii PG8 are enriched in virulence proteins.

    PubMed

    Chernov, Vladislav M; Mouzykantov, Alexey A; Baranova, Natalia B; Medvedeva, Elena S; Grygorieva, Tatiana Yu; Trushin, Maxim V; Vishnyakov, Innokentii E; Sabantsev, Anton V; Borchsenius, Sergei N; Chernova, Olga A

    2014-10-14

    Mycoplasmas (class Mollicutes), the smallest prokaryotes capable of self-replication, as well as Archaea, Gram-positive and Gram-negative bacteria constitutively produce extracellular vesicles (EVs). However, little is known regarding the content and functions of mycoplasma vesicles. Here, we present for the first time a proteomics-based characterisation of extracellular membrane vesicles from Acholeplasma laidlawii PG8. The ubiquitous mycoplasma is widespread in nature, found in humans, animals and plants, and is the causative agent of phytomycoplasmoses and the predominant contaminant of cell cultures. Taking a proteomics approach using LC-ESI-MS/MS, we identified 97 proteins. Analysis of the identified proteins indicated that A. laidlawii-derived EVs are enriched in virulence proteins that may play critical roles in mycoplasma-induced pathogenesis. Our data will help to elucidate the functions of mycoplasma-derived EVs and to develop effective methods to control infections and contaminations of cell cultures by mycoplasmas. In the present study, we have documented for the first time the proteins in EVs secreted by mycoplasma vesicular proteins identified in this study are likely involved in the adaptation of bacteria to stressors, survival in microbial communities and pathogen-host interactions. These findings suggest that the secretion of EVs is an evolutionally conserved and universal process that occurs in organisms from the simplest wall-less bacteria to complex organisms and indicate the necessity of developing new approaches to control infects.

  2. Aerosols as a Source of Widespread Mycoplasma Contamination of Tissue Cultures1

    PubMed Central

    O'Connell, Robert C.; Wittler, Ruth G.; Faber, John E.

    1964-01-01

    Mycoplasma isolates were cultured from 15 antibiotic-free cell cultures obtained from a single laboratory. Complement-fixation tests showed that these isolates were antigenically related to each other but were unrelated to M. hominis type 1, M. hominis type 2, M. arthritidis, M. laidlawii type B, Mycoplasma sp. H.Ep. #2 (Barile), or M. salivarium. Examination of serum used to feed the infected cell lines revealed no Mycoplasma. Infection resulting from cross-contamination by a single Mycoplasma strain from one cell culture to another was investigated. Although the organisms were not found in the air over the work area, aerosols containing these contaminants were produced in tissue culture bottles during the trypsinization of cell monolayers. The minimal infectious dose of Mycoplasma for tissue cultures was measured, and it was determined that one organism was capable of initiating an infection in a tissue culture. The pattern of contamination and the small dose required for infection indicated that Mycoplasma contamination was spread from one tissue culture to another via aerosols. It was demonstrated that Mycoplasma can be transferred from one cell culture to another through the use of a common burette for dispensing medium. PMID:14199025

  3. [In vitro antibiotic sensitivity of French strains of Mycoplasma bovis].

    PubMed

    Poumarat, F; Martel, J L

    1989-01-01

    The in vitro activity of 15 antibiotics was tested with 30-90 Mycoplasma bovis representative strains of bovine lung pathology in France. The distribution of minimal inhibitory concentration (MIC) is homogeneous with low values for spectinomycin, lincomycin, tylosin, gentamicin and baytril, intermediate for chloramphenicol and neomycin, high for nalidixic acid, Flumequine and erythromycin. The MIC distribution is heterogeneous with intermediate values for spiramycin and tetracyclines, and high values for streptomycin. For the later antibiotics, the heterogeneity of the susceptibility suggests a mechanism of acquired resistance.

  4. Mycoplasma hominis periaortic abscess following heart-lung transplantation.

    PubMed

    Hagiya, Hideharu; Yoshida, Hisao; Yamamoto, Norihisa; Kimura, Keigo; Ueda, Akiko; Nishi, Isao; Akeda, Yukihiro; Tomono, Kazunori

    2017-06-01

    We report the first case of Mycoplasma hominis periaortic abscess after heart-lung transplantation. The absence of sternal wound infection delayed the diagnosis, but the patient successfully recovered with debridement surgeries and long-term antibiotic therapy. Owing to the difficulty in detection and the intrinsic resistance to beta-lactams, M. hominis infections are prone to being misdiagnosed and undertreated. M. hominis should be suspected in cases where conventional microbiological identification and treatment approaches fail. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  5. Arginine catabolism by Mycoplasma meleagridis and its role in pathogenesis.

    PubMed Central

    Ibrahim, A A; Yamamoto, R

    1977-01-01

    A thin-layer chromatography technique was used to study the arginine metabolism of Mycoplasma meleagridis. The technique reflected the enzyme activity of the dihydrolase pathway through detection of readily visible end products on X-ray film. Strains of M. meleagridis differing in their pathogenicity for turkeys did not vary in arginine metabolism. In addition, no significant difference was observed in plasma arginine concentrations between M. meleagridis-infected and uninfected poults. It was concluded that the pathogenesis of M. meleagridis infection in turkeys was not based on its competition with the host for arginine. Images PMID:908618

  6. Eggshell apex abnormalities associated with Mycoplasma synoviae infection in layers

    PubMed Central

    Jeon, Eun-Ok; Kim, Jong-Nyeo; Lee, Hae-Rim; Koo, Bon-Sang; Min, Kyeong-Cheol; Han, Moo-Sung; Lee, Seung-Baek; Bae, Yeon-Ji; Mo, Jong-Suk; Cho, Sun-Hyung; Lee, Chang-Hee

    2014-01-01

    Eggs exhibiting eggshell apex abnormalities (EAA) were evaluated for changes in shell characteristics such as strength, thickness, and ultrastructure. Mycoplasma synoviae (MS) infection was confirmed by serological assay along with isolation of MS from the trachea and oviduct. Changes in eggshell quality were shown to be statistically significant (p < 0.01). We also identified ultrastructural changes in the mammillary knob layer by Scanning Electron Microscopy. While eggs may seem to be structurally sound, ultrastructural evaluation showed that affected eggs do not regain their former quality. In our knowledge, this is the first report describing the occurrence of EAA in Korea. PMID:24962418

  7. Postepizootic Persistence of Asymptomatic Mycoplasma conjunctivae Infection in Iberian Ibex.

    PubMed

    Fernández-Aguilar, Xavier; Cabezón, Oscar; Granados, José Enrique; Frey, Joachim; Serrano, Emmanuel; Velarde, Roser; Cano-Manuel, Francisco Javier; Mentaberre, Gregorio; Ráez-Bravo, Arián; Fandos, Paulino; López-Olvera, Jorge Ramón

    2017-08-01

    The susceptibility of the Iberian ibex (Capra pyrenaica) to Mycoplasma conjunctivae ocular infection and the changes in their interaction over time were studied in terms of clinical outcome, molecular detection, and IgG immune response in a captive population that underwent a severe infectious keratoconjunctivitis (IKC) outbreak. Mycoplasma conjunctivae was detected in the Iberian ibex, coinciding with the IKC outbreak. Its prevalence had a decreasing trend in 2013 that was consistent with the clinical resolution (August, 35.4%; September, 8.7%; November, 4.3%). Infections without clinical outcome were, however, still detected in the last handling in November. Sequencing and cluster analyses of the M. conjunctivae strains found 1 year later in the ibex population confirmed the persistence of the same strain lineage that caused the IKC outbreak but with a high prevalence (75.3%) of mostly asymptomatic infections and with lower DNA load of M. conjunctivae in the eyes (mean quantitative PCR [qPCR] cycle threshold [CT ], 36.1 versus 20.3 in severe IKC). Significant age-related differences of M. conjunctivae prevalence were observed only under IKC epizootic conditions. No substantial effect of systemic IgG on M. conjunctivae DNA in the eye was evidenced with a linear mixed-models selection, which indicated that systemic IgG does not necessarily drive the resolution of M. conjunctivae infection and does not explain the epidemiological changes observed. The results show how both epidemiological scenarios, i.e., severe IKC outbreak and mostly asymptomatic infections, can consecutively occur by entailing mycoplasma persistence.IMPORTANCEMycoplasma infections are reported in a wide range of epidemiological scenarios that involve severe disease to asymptomatic infections. This study allows a better understanding of the transition between two different Mycoplasma conjunctivae epidemiological scenarios described in wild host populations and highlights the ability of M

  8. Occurrence and Relevance of Mycoplasma sturni in Free-Ranging Corvids in Germany.

    PubMed

    Ziegler, Luisa; Möller Palau-Ribes, Franca Möller; Schmidt, Liane; Lierz, Michael

    2017-01-18

    Several Mycoplasma spp. are well-known pathogens in poultry. In birds of prey, White Storks ( Ciconia ciconia ), and some waterfowl (Anatidae, Pelecanidae) species, mycoplasmas occur commonly and seem to be apathogenic or commensal and most likely belong to the physiologic microbial flora of the respiratory tract. In other bird species, such as Common Nightingales ( Luscinia megarhynchos ) and tits (Paridae), Mycoplasma spp. are absent in healthy birds. In corvids, the prevalence and role of Mycoplasma spp. in disease remains unclear. In previous studies, Mycoplasma sturni was detected in diseased corvids; however, those studies included only a limited sample size or preselected individuals. We collected tracheal swabs of 97 free-ranging Corvidae, including 68 randomly selected individuals from hunting bags and 29 birds that had been admitted to a veterinary clinic. Tracheal swabs were examined for Mycoplasma spp. using culture and genus-specific PCR. If Mycoplasma spp. were detected, the species were identified by sequencing the 16S ribosomal (r)RNA gene and 16-23S rRNA intergenic transcribed spacer region. Five of 68 (7%) of the hunted birds and nine of 29 (31%) of the birds admitted to the veterinary clinic were PCR positive. In 13 of 14 PCR-positive samples, mycoplasmas were cultured and M. sturni was the only mycoplasmal species identified. None of the positive corvids from the hunting bags had clinical signs, whereas five of nine birds admitted to the veterinary clinic showed apathy, lameness, injuries, or fractures, which may not be associated with mycoplasmal infections. These data support the notion that M. sturni is the Mycoplasma sp. most frequently found in corvids, though its prevalence and ability to cause disease may involve interaction with other aspects of bird health.

  9. Pilot study to evaluate the role of Mycoplasma species in cat bite abscesses.

    PubMed

    Torres-Henderson, Camille; Hesser, Jeff; Hyatt, Doreene R; Hawley, Jennifer; Brewer, Melissa; Lappin, Michael R

    2014-12-01

    Mycoplasma species are common inhabitants of the feline oral cavity, and so likely contaminate many cat bite abscesses. The objectives of this study were to determine whether Mycoplasma species are common contaminants of cat bite abscesses and whether they are are associated with β-lactam-resistant clinical disease. Twenty-six privately owned cats with clinical evidence of an abscess suspected to be from a cat bite were included in the study. Samples from each cat were evaluated by aerobic and anaerobic culture, as well as Mycoplasma species culture and polymerase chain reaction (PCR). All cats were initially treated with appropriate wound management and were administered an antibiotic of the β-lactam class (amoxicillin, amoxicillin clavulanate or cefovecin sodium). Mycoplasma species DNA was amplified by PCR from 4/26 samples (15.4%); one of these cases was concurrently culture positive. Adequate DNA for sequencing was present for 2/4 positive PCR samples; one was most homologous with Mycoplasma felis, and the other was most homologous with Mycoplasma equigenitalium and Mycoplasma elephantis. Of the 26 cats, 25 responded to the initial treatment by day 7. The cat that failed initial treatment was positive for M equigenitalium or M elephantis DNA on days 0 and 12, and ultimately responded to administration of enrofloxacin and clindamycin. The results suggest that while Mycoplasma species can contaminate cat bite abscesses, routine wound management and β-lactam antibiotic therapy is adequate for treatment in most cases of abscess. However, as Mycoplasma species infections do not respond to β-lactam class antibiotic therapy, these organisms should be on the differential list for cats with abscesses that fail treatment with this antibiotic class.

  10. Performance of PCR-based and Bioluminescent assays for mycoplasma detection.

    PubMed

    Falagan-Lotsch, Priscila; Lopes, Talíria Silva; Ferreira, Nívea; Balthazar, Nathália; Monteiro, Antônio M; Borojevic, Radovan; Granjeiro, José Mauro

    2015-11-01

    Contaminated eukaryotic cell cultures are frequently responsible for unreliable results. Regulatory entities request that cell cultures must be mycoplasma-free. Mycoplasma contamination remains a significant problem for cell cultures and may have an impact on biological analysis since they affect many cell parameters. The gold standard microbiological assay for mycoplasma detection involves laborious and time-consuming protocols. PCR-based and Bioluminescent assays have been considered for routine cell culture screening in research laboratories since they are fast, easy and sensitive. Thus, the aim of this work is to compare the performance of two popular commercial assays, PCR-based and Bioluminescent assays, by assessing the level of mycoplasma contamination in cell cultures from Rio de Janeiro Cell Bank (RJCB) and also from customers' laboratories. The results obtained by both performed assays were confirmed by scanning electron microscopy. In addition, we evaluated the limit of detection of the PCR kit under our laboratory conditions and the storage effects on mycoplasma detection in frozen cell culture supernatants. The performance of both assays for mycoplasma detection was not significantly different and they showed very good agreement. The Bioluminescent assay for mycoplasma detection was slightly more dependable than PCR-based due to the lack of inconclusive results produced by the first technique, especially considering the ability to detect mycoplasma contamination in frozen cell culture supernatants. However, cell lines should be precultured for four days or more without antibiotics to obtain safe results. On the other hand, a false negative result was obtained by using this biochemical approach. The implementation of fast and reliable mycoplasma testing methods is an important technical and regulatory issue and PCR-based and Bioluminescent assays may be good candidates. However, validation studies are needed.

  11. MIB–MIP is a mycoplasma system that captures and cleaves immunoglobulin G

    PubMed Central

    Arfi, Yonathan; Minder, Laetitia; Di Primo, Carmelo; Le Roy, Aline; Ebel, Christine; Coquet, Laurent; Claverol, Stephane; Vashee, Sanjay; Jores, Joerg; Blanchard, Alain; Sirand-Pugnet, Pascal

    2016-01-01

    Mycoplasmas are “minimal” bacteria able to infect humans, wildlife, and a large number of economically important livestock species. Mycoplasma infections include a spectrum of clinical manifestations ranging from simple fever to fulminant inflammatory diseases with high mortality rates. These infections are mostly chronic, suggesting that mycoplasmas have developed means to evade the host immune response. Here we present and functionally characterize a two-protein system from Mycoplasma mycoides subspecies capri that is involved in the capture and cleavage of IgG. The first component, Mycoplasma Ig binding protein (MIB), is an 83-kDa protein that is able to tightly bind to the Fv region of a wide range of IgG. The second component, Mycoplasma Ig protease (MIP), is a 97-kDa serine protease that is able to cleave off the VH domain of IgG. We demonstrate that MIB is necessary for the proteolytic activity of MIP. Cleavage of IgG requires a sequential interaction of the different partners of the system: first MIB captures the IgG, and then MIP is recruited to the MIB–IgG complex, enabling protease activity. MIB and MIP are encoded by two genes organized in tandem, with homologs found in the majority of pathogenic mycoplasmas and often in multiple copies. Phylogenetic studies suggest that genes encoding the MIB–MIP system are specific to mycoplasmas and have been disseminated by horizontal gene transfer. These results highlight an original and complex system targeting the host immunoglobulins, playing a potentially key role in the immunity evasion by mycoplasmas. PMID:27114507

  12. Detecting the Diversity of Mycoplasma and Ureaplasma Endosymbionts Hosted by Trichomonas vaginalis Isolates

    PubMed Central

    Ioannidis, Anastasios; Papaioannou, Panagiota; Magiorkinis, Emmanouil; Magana, Maria; Ioannidou, Vasiliki; Tzanetou, Konstantina; Burriel, Angeliki R.; Tsironi, Maria; Chatzipanagiotou, Stylianos

    2017-01-01

    Objectives: The symbiosis of Trichomonas vaginalis and Mycoplasma hominis is the first described association between two obligate human parasites. Trichomonas is the niche and the vector for the transmission of M. hominis infection. This clinically significant symbiosis may affect T. vaginalis virulence and susceptibility to treatment. The aims of this study were to investigate the intracellularly present Mycoplasma and Ureaplasma species in T. vaginalis strains isolated from the vaginal discharge of infected women as well as to trace the diversity pattern among the species detected in the isolated strains. Methods: Hundred pure T. vaginalis cultures were isolated from ~7,500 patient specimens presented with clinical purulent vaginitis. PCR and sequencing for Mycoplasma/Ureaplasma spp. were performed in DNA extracted from the pure cultures. In addition, vaginal discharge samples were cultured for the presence of M. hominis and U. urealyticum. Phylogenetic analysis assisted the identification of interspecies relationships between the Mycoplasma and Ureaplasma isolates. Results: Fifty four percentage of T. vaginalis isolates were harboring Mycoplasma spp. Phylogenetic analysis revealed three distinct clusters, two with already characterized M. hominis and Ureaplasma spp. (37% of total Mycoplasma spp.), whereas one group formed a distinct cluster matched with the newly identified species Candidatus Mycoplasma girerdii (59.3%) and one or more unknown Mycoplasma spp. (3.7%). Conclusions: T. vaginalis strains associated with vaginal infection might host intracellular mycoplasmas or ureaplasmas. Intracellular Mollicutes that remain undetected in the extracellular environment when conventional diagnostic methods are implemented may comprise either novel species, such as Candidatus M. giredii, or unknown species with yet unexplored clinical significance. PMID:28702014

  13. Detection of Mycoplasma spp., herpesviruses, topiviruses, and ferlaviruses in samples from chelonians in Europe.

    PubMed

    Kolesnik, Ekaterina; Obiegala, Anna; Marschang, Rachel E

    2017-07-01

    We tested samples from 1,015 chelonians in Europe for Mycoplasma spp., herpesviruses, ranaviruses, picornaviruses, and ferlaviruses by PCR. Mycoplasma spp. were detected in 42.1% and herpesviruses were detected in 8.0% of tested chelonians. Differentiation of the herpesviruses revealed that 46.9% of the detected chelonian viruses were testudinid herpesvirus 1 (TeHV-1) and 54.3% were TeHV-3, including co-detections of TeHV-1 and -3 in 3 tortoises. TeHV-4 was detected in a leopard tortoise ( Stigmochelys pardalis), and a herpesvirus that could not be further characterized was found in a pond slider ( Trachemys scripta). Picornaviruses (topiviruses) were detected in 2.2% of the tested animals; ferlaviruses were found in 0.6%; no ranaviruses were detected in any of the animals tested. Mycoplasma spp. were detected significantly more often in Horsfield's tortoises ( Testudo horsfieldii), leopard tortoises, and Indian star tortoises ( Geochelone elegans) than in other species. Horsfield's tortoises were also significantly more often positive for TeHV-1. Mycoplasma and TeHV-1 were co-detected in 3.0%, and mycoplasma and TeHV-3 in 2.3%. The TeHV-4-positive tortoise was also positive for mycoplasma. Mycoplasma and picornaviruses were co-detected in 1.2% of the tortoises. A spur-thighed tortoise ( Testudo graeca) was positive for mycoplasma and a ferlavirus. In some cases, >2 pathogens were detected. A significant correlation between mycoplasma and herpesvirus detection was found. Of all tested animals, 47.6% were positive for at least one pathogen, demonstrating the importance of pathogen detection in captive chelonians.

  14. Genital mycoplasmas in women attending the Yaoundé University Teaching Hospital in Cameroon

    PubMed Central

    Njunda, Anna L.; Nsagha, Dickson S.; Assob, Jules C.N.; Palle, John N.; Kamga, Henri L.; Nde, Peter F.; Ntube, Mengang N.C.; Weledji, Patrick E.

    2011-01-01

    Genital mycoplasmas are implicated in pelvic inflammatory diseases, puerperal infection, septic abortions, low birth weight, nongonococcal urethritis and prostatitis as well as spontaneous abortion and infertility in women. There is paucity of data on colonisation of genital mycoplasma in women and their drug sensitivity patterns. The aim of our study was to determine the prevalence of genital mycoplasmas (Ureaplasma urealiticum and Mycoplasma hominis) infection and their drug sensitivity patterns in women. A mycofast kit was used for biochemical determination of mycoplasma infection in 100 randomly selected female patients aged 19–57 years, attending the University of Yaoundé Teaching Hospital (UYTH) from March to June 2010. Informed consent was sought and gained before samples were collected. Genital mycoplasmas were found in 65 patients (65%) [95% CI=55.7–74.3%] and distributed as 41 (41%) [95% CI=31.4–50.6%] for U. urealiticum and 4 (4%) [95% CI=0.20– 7.8%] for M. hominis while there was co-infection in 20 women (20%) [95% CI=12.16–27.84%]. In our study, 57 (57%) [95% CI=47.3–67%] had other organisms, which included C. albicans (19 [19%]), G. vaginalis (35 [35%]) and T. vaginalis (3 [3%]). Among the 65 women with genital mycoplasma, the highest co-infection was with G. vaginalis (33.8%). Pristinamycine was the most effective antibiotic (92%) and sulfamethoxazole the most resistant (8%) antibiotic to genital mycoplasmas. We conclude that genital mycoplasma is a problem in Cameroon and infected women should be treated together with their partners. PMID:28299057

  15. Dendritic Cells Are the Major Antigen Presenting Cells in Inflammatory Lesions of Murine Mycoplasma Respiratory Disease

    PubMed Central

    Sun, Xiangle; Jones, Harlan P.; Dobbs, Nicole; Bodhankar, Sheetal; Simecka, Jerry W.

    2013-01-01

    Mycoplasmas cause chronic respiratory diseases in animals and humans, and to date, development of vaccines have been problematic. Using a murine model of mycoplasma pneumonia, lymphocyte responses, specifically T cells, were shown to confer protection as well as promote immunopathology in mycoplasma disease. Because T cells play such a critical role, it is important to define the role of antigen presenting cells (APC) as these cells may influence either exacerbation of mycoplasma disease pathogenesis or enhancement of protective immunity. The roles of APC, such as dendritic cells and/or macrophages, and their ability to modulate adaptive immunity in mycoplasma disease are currently unknown. Therefore, the purpose of this study was to identify individual pulmonary APC populations that may contribute to the activation of T cell responses during mycoplasma disease pathogenesis. The present study indeed demonstrates increasing numbers of CD11c− F4/80+ cells, which contain macrophages, and more mature/activated CD11c+ F4/80− cells, containing DC, in the lungs after infection. CD11c− F4/80+ macrophage-enriched cells and CD11c+ F4/80− dendritic cell-enriched populations showed different patterns of cytokine mRNA expression, supporting the idea that these cells have different impacts on immunity in response to infection. In fact, DC containing CD11c+ F4/80− cell populations from the lungs of infected mice were most capable of stimulating mycoplasma-specific CD4+ Th cell responses in vitro. In vivo, these CD11c+F4/80− cells were co-localized with CD4+ Th cells in inflammatory infiltrates in the lungs of mycoplasma-infected mice. Thus, CD11c+F4/80− dendritic cells appear to be the major APC population responsible for pulmonary T cell stimulation in mycoplasma-infected mice, and these dendritic cells likely contribute to responses impacting disease pathogenesis. PMID:23390557

  16. Rapid imaging of mycoplasma in solution using Atmospheric Scanning Electron Microscopy (ASEM)

    SciTech Connect

    Sato, Chikara; Manaka, Sachie; Nakane, Daisuke; Nishiyama, Hidetoshi; Suga, Mitsuo; Nishizaka, Takayuki; Miyata, Makoto; Maruyama, Yuusuke

    2012-01-27

    Highlights: Black-Right-Pointing-Pointer Mycoplasma mobile was observed in buffer with the Atmospheric Scanning Electron Microscope. Black-Right-Pointing-Pointer Characteristic protein localizations were visualized using immuno-labeling. Black-Right-Pointing-Pointer M. mobile attached to sialic acid on the SiN film surface within minutes. Black-Right-Pointing-Pointer Cells were observed at low concentrations. Black-Right-Pointing-Pointer ASEM should promote study and early-stage diagnosis of mycoplasma. -- Abstract: Mycoplasma is a genus of bacterial pathogen that causes disease in vertebrates. In humans, the species Mycoplasma pneumoniae causes 15% or more of community-acquired pneumonia. Because this bacterium is tiny, corresponding in size to a large virus, diagnosis using optical microscopy is not easy. In current methods, chest X-rays are usually the first action, followed by serology, PCR amplification, and/or culture, but all of these are particularly difficult at an early stage of the disease. Using Mycoplasma mobile as a model species, we directly observed mycoplasma in buffer with the newly developed Atmospheric Scanning Electron Microscope (ASEM). This microscope features an open sample dish with a pressure-resistant thin film window in its base, through which the SEM beam scans samples in solution, from below. Because of its 2-3 {mu}m-deep scanning capability, it can observe the whole internal structure of mycoplasma cells stained with metal solutions. Characteristic protein localizations were visualized using immuno-labeling. Cells were observed at low concentrations, because suspended cells concentrate in the observable zone by attaching to sialic acid on the silicon nitride (SiN) film surface within minutes. These results suggest the applicability of the ASEM for the study of mycoplasmas as well as for early-stage mycoplasma infection diagnosis.

  17. Detection of multiple Mycoplasma species in bulk tank milk samples using real-time PCR and conventional culture and comparison of test sensitivities.

    PubMed

    Justice-Allen, A; Trujillo, J; Goodell, G; Wilson, D

    2011-07-01

    The objective of this study was to further validate a SYBR PCR protocol for Mycoplasma spp. by comparing it with standard microbial culture in the detection of Mycoplasma spp. in bulk tank milk samples. Additionally, we identified Mycoplasma spp. present by analysis of PCR-generated amplicons [dissociation (melt) temperature (T(m)), length, and DNA sequence]. The research presented herein tests the hypothesis that the SYBR PCR protocol is as sensitive as conventional culture for the detection of Mycoplasma spp. in bulk tank milk samples. Mycoplasmas cause several important disease syndromes in cattle, including mastitis in dairy cows. The standard diagnostic method at the herd level has been microbial isolation of mycoplasmas on 1 of several specialized media and speciation through biochemical or immunological techniques; repeated sampling schemes are recommended. The development of a real-time SYBR PCR protocol offers advantages in decrease of time to detection, cost, and complexity. The T(m) of the double-stranded DNA generated from the PCR reaction was used to detect the presence of and tentatively identify the species of mycoplasmas other than Mycoplasma bovis. In the SYBR PCR protocol, the presence of multiple species of mycoplasmas is indicated by an atypical dissociation curve. Gel electrophoresis and sequencing of the amplicons was used to confirm the mycoplasma species present when a non-M. bovis organism was detected (T(m) not equal to M. bovis) and used to identify all the mycoplasma species present for the samples with atypical dissociation curves. Mycoplasma bovis was identified in 83% of SYBR PCR mycoplasma-positive bulk tank samples. Another mycoplasma was identified either alone or in addition to M. bovis in 25% of SYBR PCR mycoplasma-positive bulk tank milk samples. Four species of mycoplasma other than M. bovis (Mycoplasma alkalescens, Mycoplasma arginini, Mycoplasma bovigenitalium, and Mycoplasma gateae) were identified in bulk tank milk samples

  18. Short communication: The effect of centrifugation and resuspension on the recovery of Mycoplasma species from milk.

    PubMed

    Punyapornwithaya, V; Fox, L K; Gay, G M; Hancock, D D; Alldredge, J R

    2009-09-01

    Low sensitivity of a single bulk tank milk culture is a major limitation for detection of mycoplasma organisms. We hypothesized that sedimentation of Mycoplasma spp. in a milk sample by centrifugation followed by resuspension in a small volume of fluid before agar plating would increase the ability to detect Mycoplasma spp. compared with direct conventional culture. The experiment was conducted to determine recovery of Mycoplasma spp. from milk as affected by 1) treatment (centrifugation vs. conventional method); 2) 2 species (Mycoplasma bovis and Mycoplasma californicum and 4 strains for each species); and 3) 4 different concentrations of Mycoplasma spp. (1,000, 100, 10, and 1 cfu/mL). A 5-mL portion of mycoplasma suspension from each strain was inoculated into 45 mL of fresh bulk tank milk to achieve concentrations of 1,000, 100, 10, and 1 cfu/mL. Treatment samples were vigorously mixed and centrifuged at 5,000 x g for 30 min. Control samples were vigorously mixed. All samples were plated on modified Hayflick agar. Plates were incubated at 37 degrees C and 5% CO(2) for 5 d. Mean (+/-SE) log(10) mycoplasma counts (cfu/mL) in the treatment groups (1.91 +/- 0.15) were higher than those in the control groups (1.70 +/- 0.16). Recovery of at least 1 mycoplasma colony on agar culture was 100% in both treatment and control groups at high, medium, and low concentrations. At the lowest concentration, recovery of at least 1 mycoplasma colony on agar culture in treatment and control groups was 75% (n = 12/16) and 18.75% (n = 3/16), respectively. Centrifugation of milk followed by suspension in a smaller volume of saline before conventional culture increased the ability to detect mycoplasma microorganisms in the milk sample compared with controls. Recovery by centrifugation appeared best at the lowest concentration where detection of a positive sample was 4 times more likely than when conventional methods were used.

  19. Anti-Gal-C antibody in autoimmune neuropathies subsequent to mycoplasma infection.

    PubMed

    Kusunoki, S; Chiba, A; Hitoshi, S; Takizawa, H; Kanazawa, I

    1995-04-01

    Four of 82 patients with Guillain-Barré syndrome (GBS) and 1 of 12 with multifocal motor neuropathy (MMN), who previously had had Mycoplasma pneumoniae infections, had serum antibody to galactocerebroside (Gal-C). Two patients with GBS without mycoplasma infection also had anti-Gal-C antibody, whereas none of the normal or the disease controls had it. As Gal-C is a major glycolipid antigen in myelin, anti-Gal-C antibody may function in the pathogenesis of autoimmune demyelinative neuropathies. Mycoplasma pneumoniae appears to be an important preceding infectious agent in autoimmune neuropathies with anti-Gal-C antibody.

  20. Nasal prevalence of Mycoplasma bovis and IHA titers in young dairy animals.

    PubMed

    Bennett, R H; Jasper, D E

    1977-07-01

    Serologic and cultural observations were made in three herds with and three herds without histories of mycoplasma mastitis. Nasal swabs and sera were collected from dairy animals of various ages over an eight month peiod. The overall prevalence of Myocopalsma bovis in the nares was 34% in diseased herds and 6% in the non-diseased herds without mastitis. Mycoplasma bovis was isolated in the highest prevalence in those young animals fed infected milk. Slight serologic differences were seen in these animals. Nasal prevalence of M. bovis was low but readily detectable in non diseased herds as well as in prepartum heifers in the diseased herds with mycoplasma mastitis.

  1. Characterisation of mycoplasmas isolated from genital tract infections of sheep in Nigeria.

    PubMed

    Chima, J C; Ojo, M O; Molokwu, J U; Okewole, P A

    1995-09-01

    Four mycoplasma-like organisms isolated from ewes with mucopurulent vaginal discharge and swollen vulva were characterised. Biochemical tests showed three of the isolates to be negative for glucose fermentation and arginine hydrolysis, while the remaining isolate was negative for glucose fermentation but hydrolysed arginine. Serological identification using the growth inhibition, growth precipitation and indirect immunofluorescence tests indicated the three similar isolates as Mycoplasma bovigenitalium and the other isolate as Mycoplasma arginini. There are apparently no previous reports of the isolation of these organisms from the genital tract of sheep in Nigeria.

  2. Mycoplasma testing of cell substrates and biologics: Review of alternative non-microbiological techniques.

    PubMed

    Volokhov, Dmitriy V; Graham, Laurie J; Brorson, Kurt A; Chizhikov, Vladimir E

    2011-01-01

    Mycoplasmas, particularly species of the genera Mycoplasma and Acholeplasma, are known to be occasional microbial contaminants of cell cultures that produce biologics. This presents a serious concern regarding the risk of mycoplasma contamination for research laboratories and commercial facilities developing and manufacturing cell-derived biological and biopharmaceutical products for therapeutic use. Potential undetected contamination of these products or process intermediates with mycoplasmas represents a potential safety risk for patients and a business risk for producers of biopharmaceuticals. To minimize these risks, monitoring for adventitious agents, such as viruses and mycoplasmas, is performed during the manufacture of biologics produced in cell culture substrates. The "gold standard" microbiological assay, currently recommended by the USP, EP, JP and the US FDA, for the mycoplasma testing of biologics, involves the culture of viable mycoplasmas in broth, agar plates and indicator cells. Although the procedure enables highly efficient mycoplasma detection in cell substrates and cell-derived products, the overall testing strategy is time consuming (a minimum of 28 days) and requires skilled interpretation of the results. The long time period required for these conventional assays does not permit their use for products with short shelf-lives or for timely 'go/no-go' decisions during routine in-process testing. PCR methodology has existed for decades, however PCR based and other alternative methods for mycoplasma detection have only recently been considered for application to biologics manufacture. The application of alternative nucleic acid-based, enzyme-based and/or recombinant cell-culture methods, particularly in combination with efficient sample preparation procedures, could provide advantages over conventional microbiological methods in terms of analytical throughput, simplicity, and turnaround time. However, a challenge to the application of alternative

  3. Identification and Characterization of Mycoplasma feriruminatoris sp. nov. Strains Isolated from Alpine Ibex: A 4th Species in the Mycoplasma mycoides Cluster Hosted by Non-domesticated Ruminants?

    PubMed Central

    Ambroset, Chloé; Pau-Roblot, Corinne; Game, Yvette; Gaurivaud, Patrice; Tardy, Florence

    2017-01-01

    The genus Mycoplasma, a group of free-living, wall-less prokaryotes includes more than 100 species of which dozens are primary pathogens of humans and domesticated animals. Mycoplasma species isolated from wildlife are rarely investigated but could provide a fuller picture of the evolutionary history and diversity of this genus. In 2013 several isolates from wild Caprinae were tentatively assigned to a new species, Mycoplasma (M.) feriruminatoris sp. nov., characterized by an unusually rapid growth in vitro and close genetic proximity to ruminant pathogenic species. We suspected that atypical isolates recently collected from Alpine ibex in France belonged to this new species. The present study was undertaken to verify this hypothesis and to further characterize the French ibex isolates. Phylogenetic analyses were performed to identify the isolates and position them in trees containing several other mycoplasma species pathogenic to domesticated ruminants. Population diversity was characterized by genomic macrorestriction and by examining the capacity of different strains to produce capsular polysaccharides, a feature now known to vary amongst mycoplasma species pathogenic to ruminants. This is the first report of M. feriruminatoris isolation from Alpine ibex in France. Phylogenetic analyses further suggested that M. feriruminatoris might constitute a 4th species in a genetic cluster that so far contains only important ruminant pathogens, the so-called Mycoplasma mycoides cluster. A PCR assay for specific identification is proposed. These French isolates were not clonal, despite being collected in a restricted region of the Alps, which signifies a considerable diversity of the new species. Strains were able to concomitantly produce two types of capsular polysaccharides, β-(1→6)-galactan and β-(1→6)-glucan, with variation in their respective ratio, a feature never before described in mycoplasmas. PMID:28611743

  4. Life on arginine for Mycoplasma hominis: clues from its minimal genome and comparison with other human urogenital mycoplasmas.

    PubMed

    Pereyre, Sabine; Sirand-Pugnet, Pascal; Beven, Laure; Charron, Alain; Renaudin, Hélène; Barré, Aurélien; Avenaud, Philippe; Jacob, Daniel; Couloux, Arnaud; Barbe, Valérie; de Daruvar, Antoine; Blanchard, Alain; Bébéar, Cécile

    2009-10-01

    Mycoplasma hominis is an opportunistic human mycoplasma. Two other pathogenic human species, M. genitalium and Ureaplasma parvum, reside within the same natural niche as M. hominis: the urogenital tract. These three species have overlapping, but distinct, pathogenic roles. They have minimal genomes and, thus, reduced metabolic capabilities characterized by distinct energy-generating pathways. Analysis of the M. hominis PG21 genome sequence revealed that it is the second smallest genome among self-replicating free living organisms (665,445 bp, 537 coding sequences (CDSs)). Five clusters of genes were predicted to have undergone horizontal gene transfer (HGT) between M. hominis and the phylogenetically distant U. parvum species. We reconstructed M. hominis metabolic pathways from the predicted genes, with particular emphasis on energy-generating pathways. The Embden-Meyerhoff-Parnas pathway was incomplete, with a single enzyme absent. We identified the three proteins constituting the arginine dihydrolase pathway. This pathway was found essential to promote growth in vivo. The predicted presence of dimethylarginine dimethylaminohydrolase suggested that arginine catabolism is more complex than initially described. This enzyme may have been acquired by HGT from non-mollicute bacteria. Comparison of the three minimal mollicute genomes showed that 247 CDSs were common to all three genomes, whereas 220 CDSs were specific to M. hominis, 172 CDSs were specific to M. genitalium, and 280 CDSs were specific to U. parvum. Within these species-specific genes, two major sets of genes could be identified: one including genes involved in various energy-generating pathways, depending on the energy source used (glucose, urea, or arginine) and another involved in cytadherence and virulence. Therefore, a minimal mycoplasma cell, not including cytadherence and virulence-related genes, could be envisaged containing a core genome (247 genes), plus a set of genes required for providing

  5. Life on Arginine for Mycoplasma hominis: Clues from Its Minimal Genome and Comparison with Other Human Urogenital Mycoplasmas

    PubMed Central

    Pereyre, Sabine; Sirand-Pugnet, Pascal; Beven, Laure; Charron, Alain; Renaudin, Hélène; Barré, Aurélien; Avenaud, Philippe; Jacob, Daniel; Couloux, Arnaud; Barbe, Valérie; de Daruvar, Antoine; Blanchard, Alain; Bébéar, Cécile

    2009-01-01

    Mycoplasma hominis is an opportunistic human mycoplasma. Two other pathogenic human species, M. genitalium and Ureaplasma parvum, reside within the same natural niche as M. hominis: the urogenital tract. These three species have overlapping, but distinct, pathogenic roles. They have minimal genomes and, thus, reduced metabolic capabilities characterized by distinct energy-generating pathways. Analysis of the M. hominis PG21 genome sequence revealed that it is the second smallest genome among self-replicating free living organisms (665,445 bp, 537 coding sequences (CDSs)). Five clusters of genes were predicted to have undergone horizontal gene transfer (HGT) between M. hominis and the phylogenetically distant U. parvum species. We reconstructed M. hominis metabolic pathways from the predicted genes, with particular emphasis on energy-generating pathways. The Embden–Meyerhoff–Parnas pathway was incomplete, with a single enzyme absent. We identified the three proteins constituting the arginine dihydrolase pathway. This pathway was found essential to promote growth in vivo. The predicted presence of dimethylarginine dimethylaminohydrolase suggested that arginine catabolism is more complex than initially described. This enzyme may have been acquired by HGT from non-mollicute bacteria. Comparison of the three minimal mollicute genomes showed that 247 CDSs were common to all three genomes, whereas 220 CDSs were specific to M. hominis, 172 CDSs were specific to M. genitalium, and 280 CDSs were specific to U. parvum. Within these species-specific genes, two major sets of genes could be identified: one including genes involved in various energy-generating pathways, depending on the energy source used (glucose, urea, or arginine) and another involved in cytadherence and virulence. Therefore, a minimal mycoplasma cell, not including cytadherence and virulence-related genes, could be envisaged containing a core genome (247 genes), plus a set of genes required for providing

  6. Macromolecular Synthesis and Thymineless Death in Mycoplasma laidlawii B1

    PubMed Central

    Smith, Douglas W.; Hanawalt, Philip C.

    1968-01-01

    The relationships between macromolecular synthesis and viability have been studied in the pleuropneumonia-like organism Mycoplasma laidlawii B adapted to a semidefined grwoth medium. This organism exhibited an absolute growth requirement for the nucleosides uridine and thymidine, a partial requirement for guanosine and deoxyguanosine, but no requirement for adenosine, deoxyadenosine, cytosine, and deoxycytosine. Cytosine and deoxycytosine partially satisfied the requirement for uridine. Loss in viability resulted from thymidine deprivation, but not from a deficiency in other growth requirements. This phenomenon of thymineless death in a mycoplasma is similar in many respects to that reported in other bacterial systems. Chloramphenicol specifically inhibited protein synthesis and allowed deoxyribonucleic acid synthesis to proceed to only about 40% of that normally produced per generation period, while causing less inhibition of ribonucleic acid synthesis. Protein synthesis inhibition permitted thymineless death to a survival level of less than 0.5%, but ribonucleic acid synthesis inhibition resulted in a higher (10%) survival level. These results are consistent with previously noted aspects of thymineless death in Escherichia coli strains, which suggest that thymineless death is coupled to ribonucleic acid synthesis. PMID:4881702

  7. Global transcriptional analysis of Mycoplasma hyopneumoniae following exposure to norepinephrine.

    PubMed

    Oneal, Michael J; Schafer, Erin R; Madsen, Melissa L; Minion, F Chris

    2008-09-01

    Mycoplasma hyopneumoniae, a component of the porcine respiratory disease complex, colonizes the respiratory tract of swine by binding to the cilia of the bronchial epithelial cells. Mechanisms of pathogenesis are poorly understood for M. hyopneumoniae, but previous work has indicated that it responds to the environmental stressors heat shock, iron deprivation and oxidative compounds. For successful infection, M. hyopneumoniae must effectively resist host responses to the colonization of the respiratory tract. Among these are changes in hormonal levels in the mucosal secretions. Recent work in the stress responses of other bacteria has included the response to the catecholamine norepinephrine. The idea that M. hyopneumoniae can respond to a host hormone, however, is novel and has not previously been demonstrated. To test this, organisms in the early exponential phase of growth were exposed to 100 muM norepinephrine for 4 h, and RNA samples from these cultures were collected and compared to RNA samples from control cultures using two-colour PCR-based M. hyopneumoniae microarrays. The M. hyopneumoniae response included slowed growth and changes in mRNA transcript levels of 84 genes, 53 of which were upregulated in response to norepinephrine. A larger proportion of the genes upregulated than those downregulated were involved with transcription and translation. The downregulated genes were mostly involved with metabolism, which correlated with the reduction in growth of the mycoplasma. Approximately 51 % of the genes were hypothetical with no known function. Thus, in response to norepinephrine, M. hyopneumoniae appears to upregulate protein expression while downregulating general metabolism.

  8. Clinical Features of Severe or Fatal Mycoplasma pneumoniae Pneumonia

    PubMed Central

    Izumikawa, Koichi

    2016-01-01

    Mycoplasma pneumoniae is one of the most common causes of community-acquired pneumonia in children and young adults. The incidence of fulminant M. pneumoniae pneumonia (MPP) is relatively rare despite the high prevalence of M. pneumoniae infection. This literature review highlights the clinical features of fulminant MPP by examining the most recent data in epidemiology, clinical presentation, pathogenesis, and treatment. Fulminant MPP accounts for 0.5–2% of all MPP cases and primarily affects young adults with no underlying disease. Key clinical findings include a cough, fever, and dyspnea along with diffuse abnormal findings in radiological examinations. Levels of inflammatory markers such as white blood cells and C-reactive protein are elevated, as well as levels of lactate dehydrogenase, IL-18, aspartate transaminase, and alanine transaminase. The exact pathogenesis of fulminant MPP remains unclear, but theories include a delayed hypersensitivity reaction to M. pneumoniae and the contribution of delayed antibiotic administration to disease progression. Treatment options involve pairing the appropriate anti-mycoplasma agent with a corticosteroid that will downregulate the hypersensitivity response, and mortality rates are quite low in this treatment group. Further research is necessary to determine the exact pathogenesis of severe and fulminant types of MPP. PMID:27313568

  9. First isolation of Mycoplasma iowae in grey partridge flocks.

    PubMed

    Catania, S; Gobbo, F; Rodio, S; Qualtieri, K; Santone, C; Nicholas, R A J

    2014-06-01

    Mycoplasma iowae, an occasional pathogen of turkeys, was isolated for the first time from captive grey partridges (Perdix perdix). Clinical signs including respiratory and intestinal disorder were seen in birds of all ages but mainly in those kept housed during rearing. Mortality rates averaged over 20% during the year. Treatment with antibiotics and antiparasitic drugs produced only a transient improvement in condition. The gross pathology findings included poor body growth, lack of development of the breast muscles, abnormalities in the keel development, and bone fragility. Some birds showed infraorbital sinusitis with serous or fibrinous exudates and catarrhal tracheitis, while others presented serofibrinous airsacculitis and splenomegaly. Laboratory investigations revealed pure cultures of M. iowae in the gut as well as sinus and air sacs. While other organisms such as coccidia, Trichomonas, Escherichia coli, Clostridium perfringens, and Aspergillus spp. were detected, the similarity of the disease with that seen in turkeys infected with M. iowae strongly suggests that this mycoplasma may be the primary pathogen here. The presence of M. iowae in game birds commonly released into the wild could have serious implications particularly in areas where industrial poultry farms are concentrated.

  10. Mycoplasma agassizii in Morafka's desert tortoise (Gopherus morafkai) in Mexico.

    PubMed

    Berry, Kristin H; Brown, Mary B; Vaughn, Mercy; Gowan, Timothy A; Hasskamp, Mary Ann; Torres, Ma Cristina Meléndez

    2015-01-01

    We conducted health evaluations of 69 wild and 22 captive Morafka's desert tortoises (Gopherus morafkai) in Mexico between 2005 and 2008. The wild tortoises were from 11 sites in the states of Sonora and Sinaloa, and the captive tortoises were from the state-managed Centro Ecológico de Sonora Zoo in Hermosillo and a private residence in the town of Alamos. We tested 88 tortoises for mycoplasmal upper respiratory tract disease (URTD) using enzyme-linked immunosorbent assays for specific antibody and by culture and PCR for detection of Mycoplasma agassizii and Mycoplasma testudineum. Fifteen of 22 captive tortoises had one or more positive diagnostic test results for M. agassizii whereas no wild tortoises had positive tests. Tortoises with positive tests also had significantly more moderate and severe clinical signs of mycoplasmosis on beaks and nares compared to tortoises with negative tests. Captive tortoises also exhibited significantly more clinical signs of illness than did wild tortoises, including lethargy and moderate to severe ocular signs. The severity of trauma and diseases of the shell and integument did not differ significantly among tortoises by site; however, clinical signs of moderate to severe trauma and disease were more prevalent in older tortoises. Similar to research findings for other species in the genus Gopherus in the US, we found that URTD is an important disease in captive tortoises. If they escape or are released by intention or accident to the wild, captive tortoises are likely to pose risks to healthy, naïve wild populations.

  11. Prevalence of Mycoplasma ovipneumoniae in desert bighorn sheep in Arizona

    USGS Publications Warehouse

    Justice-Allen, Anne E.; Luedtke, Clint J.; Overstreet, Matthew; Cain, James W.; Stephenson, Thomas R.

    2011-01-01

    To assess the potential for an epizootic of pneumonia to result from either natural immigration or translocation, we compared the seroprevalence to Mycoplasma ovipneumoniae in several populations of desert bighorn sheep in Arizona. We collected blood samples and nasal or oropharyngeal swabs from 124 desert bighorn sheep (Ovis canadensis nelsoni) from 6 populations in Arizona in 2009 and 2010. M. ovipneumoniae organisms were detected by PCR in 22%, whereas antibodies to M. ovipneumoniae were detected in 47% of tested bighorn sheep. Mycoplasma antibodies were not found in 2 of 6 populations, indicating some bighorn sheep populations in Arizona are naïve to this bacterium. In contrast, others had seroprevalence rates up to 80%. We were able to compare seroprevalence rates and titers over time in 9 individuals (7 individuals included in the 124 bighorn sheep sampled in 2009 and 2010, and 2 individuals originally captured in 2006). Antibody titers persisted for 12 months in individuals from the Kofa National Wildlife Refuge (n = 7) while antibody titers appeared to decline in the Kanab Creek population (n = 2). M. ovipneumoniae is present or has been present in several, but not all, populations of bighorn sheep in Arizona. The results demonstrate the importance of routine health testing for future translocation efforts to reduce disease risk for naive populations.

  12. Urethral infection in male chimpanzees produced experimentally by Mycoplasma genitalium.

    PubMed Central

    Taylor-Robinson, D.; Tully, J. G.; Barile, M. F.

    1985-01-01

    Four young male chimpanzees were inoculated intra-urethrally with a strain (G37) of Mycoplasma genitalium which had been isolated from the urethra of a patient with non-gonococcal urethritis. Two of the chimpanzees became infected as indicated by persistent recovery of the organisms from the urethra for 13 weeks and by an antibody response measured by both metabolism inhibition and micro-immunofluorescence techniques. The numbers of organisms isolated from both animals increased about 4 weeks after inoculation and antibody development was first detected 1 week later. The infected animals developed a minimal and inconsistently detected urethral polymorphonuclear leucocyte response which was not seen in those that were uninfected, nor in a chimpanzee that had been given medium only. The organisms were not isolated and the cellular response was not observed after treatment of the infected chimpanzees with oxytetracycline. One of the animals that had been infected was re-inoculated with strain G37 six months after successful treatment, but although the titre of serum antibody had diminished to its original level urethral recolonization did not occur. The organisms in the inoculum were not attenuated, however, because they infected another chimpanzee that had not had previous experience of M. genitalium. The results are discussed in relation to the potential of this mycoplasma to produce urethritis in man. PMID:3970830

  13. Bovine mastitis in Ontario due to Mycoplasma agalactiae subsp. bovis.

    PubMed Central

    Ruhnke, H L; Thawley, D; Nelson, F C

    1976-01-01

    Bovine mastitis caused by Mycoplasma agalactiae subsp. bovis was first diagnosed in 16 of 55 cows in an Ontario herd in Feburary 1972. A total of 182 of 598 (30.4%) cows from 33 of 64 (51.5%) farms in widely separated areas of the province were culturally positive. Herd incidence varied from 15 to 40% with one closed herd having an incidence of 61%. Four herds were investigated culturally and serologically by the growth inhibition test for 15 months. In the acute phase the organism was present in the milk in extremely high numbers and could still be isolated from a few cows after eight to 12 months. The sera from 89.5% of the animals with clinical mycoplasma mastitis produced a zone of surface "film" and/or colony inhibition and some cows remained positive for six to 12 months. The disease was experimentally reproduced with a pure culture of the organism isolated from the milk of a cow from one of the herds. PMID:1000385

  14. Cloning, Stability, and Modification of Mycoplasma hominis Genome in Yeast.

    PubMed

    Rideau, Fabien; Le Roy, Chloé; Descamps, Elodie C T; Renaudin, Hélène; Lartigue, Carole; Bébéar, Cécile

    2017-05-19

    Mycoplasma hominis is a minimal human pathogen that is responsible for genital and neonatal infections. Despite many attempts, there is no efficient genetic tool to manipulate this bacterium, limiting most investigations of its pathogenicity and its uncommon energy metabolism that relies on arginine. The recent cloning and subsequent engineering of other mycoplasma genomes in yeast opens new possibilities for studies of the genomes of genetically intractable organisms. Here, we report the successful one-step cloning of the M. hominis PG21 genome in yeast using the transformation-associated recombination (TAR) cloning method. At low passages, the M. hominis genome cloned into yeast displayed a conserved size. However, after ∼60 generations in selective media, this stability was affected, and large degradation events were detected, raising questions regarding the stability of large heterologous DNA molecules cloned in yeast and the need to minimize host propagation. Taking these results into account, we selected early passage yeast clones and successfully modified the M. hominis PG21 genome using the CRISPR/Cas9 editing tool, available in Saccharomyces cerevisiae. Complete M. hominis PG21 genomes lacking the adhesion-related vaa gene were efficiently obtained.

  15. Further observations on the murine model of Mycoplasma hominis infection.

    PubMed

    Taylor-Robinson, David; Furr, Patricia M

    2010-08-01

    Mycoplasma hominis, the first mycoplasma of human origin to be isolated, has been associated with several diseases, notably bacterial vaginosis, pelvic inflammatory disease, prematurity and puerperal fever. The mouse model does not mimic closely these features of human disease, but has some notable features. Given intravaginally to mice, M. hominis does not colonize unless the mice have been pre-treated with oestradiol. As shown here, endogenous hormone has no part to play because removal of the ovaries does not interfere with vaginal colonization. Persistent colonization occurs in hysterectomized mice so that organisms in the upper tract, which are sometimes found, are not responsible, by retrograde leakage, for those in the lower tract. Organisms in the lower tract can be eliminated by treating mice with a tetracycline, or progesterone or by natural resolution. Elimination by whatever means results in a rather weak immunity to recolonization. In contrast, intravenous inoculation of viable, and particularly killed, M. hominis organisms results in strong resistance to recolonization. This is, in part, genetically influenced, being seen in mice of strain BALB/c but not of strain CBA. Resistance is inversely proportional to the presence and titre of M. hominis specific serum antibody. The possible role of cell-mediated immunity is discussed.

  16. Association of Mycoplasma hominis infection with prostate cancer

    PubMed Central

    Barykova, Yulia A.; Logunov, Denis Yu.; Shmarov, Maxim M.; Vinarov, Andrei Z.; Fiev, Dmitry N.; Vinarova, Natalia A.; Rakovskaya, Irina V.; Baker, Patricia Stanhope; Shyshynova, Inna; Stephenson, Andrew J.; Klein, Eric A.; Naroditsky, Boris S.; Gintsburg, Alexander L.; Gudkov, Andrei V.

    2011-01-01

    The origin of chronic inflammation preceding the development of prostate cancer (PCa) remains unknown. We investigated possible involvement of mycoplasma infection in PCa by screening prostate biopsies from two groups of Russian men undergoing PCa diagnosis. M. hominis was detected by standard PCR in 15% of the 125 patients in the first group and by quantitative real-time PCR in 37.4% of the 123 men in the second group. In both groups, stratification of patients according to diagnosis showed that M. hominis was present at three times higher frequency in patients with PCa than in those with benign prostatic hyperplasia. No M. hominis was detected in the prostates of 27 men without detectable prostate disease. In addition, PCa-positive men had higher titers of antibodies against M. hominis and average PSA levels were higher in M. hominis-positive men. These data, together with previous observations linking mycoplasma infection with cell transformation, genomic instability and resistance to apoptosis, suggest that M. hominis infection may be involved in PCa development and may, therefore, be a potential PCa marker and/or target for improved prevention and treatment of this disease. PMID:21471611

  17. Association of Mycoplasma hominis infection with prostate cancer.

    PubMed

    Barykova, Yulia A; Logunov, Denis Yu; Shmarov, Maxim M; Vinarov, Andrei Z; Fiev, Dmitry N; Vinarova, Natalia A; Rakovskaya, Irina V; Baker, Patricia Stanhope; Shyshynova, Inna; Stephenson, Andrew J; Klein, Eric A; Naroditsky, Boris S; Gintsburg, Alexander L; Gudkov, Andrei V

    2011-04-01

    The origin of chronic inflammation preceding the development of prostate cancer (PCa) remains unknown. We investigated possible involvement of mycoplasma infection in PCa by screening prostate biopsies from two groups of Russian men undergoing PCa diagnosis. M. hominis was detected by standard PCR in 15% of the 125 patients in the first group and by quantitative real-time PCR in 37.4% of the 123 men in the second group. In both groups, stratification of patients according to diagnosis showed that M. hominis was present at three times higher frequency in patients with PCa than in those with benign prostatic hyperplasia. No M. hominis was detected in the prostates of 27 men without detectable prostate disease. In addition, PCa-positive men had higher titers of antibodies against M. hominis and average PSA levels were higher in M. hominis-positive men. These data, together with previous observations linking mycoplasma infection with cell transformation, genomic instability and resistance to apoptosis, suggest that M. hominis infection may be involved in PCa development and may, therefore, be a potential PCa marker and/or target for improved prevention and treatment of this disease.

  18. Postoperative Mycoplasma hominis brain abscess: keep it in mind!

    PubMed

    Bergin, Sarah Maria; Mendis, Shehara M; Young, Barnaby; Binti Izharuddin, Ezlyn

    2017-01-09

    A temporal lobe abscess was diagnosed in a 57-year-old man. A urethral catheter had been inserted 12 days earlier, just prior to clot evacuation of a subacute haematoma secondary to an arterio-venous malformation. Fever persisted despite debridement and treatment with meropenem and vancomycin. Gram stains of operative samples showed no bacteria. Extended cultures grew pinpoint colonies after 5 days. Meanwhile, sequencing of bacterial 16S rDNA from operative specimens had identified Mycoplasma hominis; the bacterial colonies were subsequently similarly identified. The patient responded promptly following addition of oral doxycycline 100 mg two times per day. There is a growing literature of similar cases. Transient bacteraemia, following urinary catheterisation, with seeding of existing sites of inflammation is the proposed explanation. Urethral carriage of M. hominis is 15% and catheterisation is a common procedure. Mycoplasma hominis maybe more common than appreciated, especially as the need for extended cultures makes a correct diagnosis less likely. 2017 BMJ Publishing Group Ltd.

  19. Mycoplasma hyopneumoniae Transcription Unit Organization: Genome Survey and Prediction

    PubMed Central

    Siqueira, Franciele Maboni; Schrank, Augusto; Schrank, Irene Silveira

    2011-01-01

    Mycoplasma hyopneumoniae is associated with swine respiratory diseases. Although gene organization and regulation are well known in many prokaryotic organisms, knowledge on mycoplasma is limited. This study performed a comparative analysis of three strains of M. hyopneumoniae (7448, J and 232), with a focus on genome organization and gene comparison for open read frame (ORF) cluster (OC) identification. An in silico analysis of gene organization demonstrated 117 OCs and 34 single ORFs in M. hyopneumoniae 7448 and J, while 116 OCs and 36 single ORFs were identified in M. hyopneumoniae 232. Genomic comparison revealed high synteny and conservation of gene order between the OCs defined for 7448 and J strains as well as for 7448 and 232 strains. Twenty-one OCs were chosen and experimentally confirmed by reverse transcription–PCR from M. hyopneumoniae 7448 genome, validating our prediction. A subset of the ORFs within an OC could be independently transcribed due to the presence of internal promoters. Our results suggest that transcription occurs in ‘run-on’ from an upstream promoter in M. hyopneumoniae, thus forming large ORF clusters (from 2 to 29 ORFs in the same orientation) and indicating a complex transcriptional organization. PMID:22086999

  20. Experimentally induced bovine abortion with Mycoplasma agalactiae subsp bovis.

    PubMed

    Stalheim, O H; Proctor, S J

    1976-08-01

    Two pregnant cows aborted 11 and 18 days after Mycoplasma agalactiae subsp bovis was inoculated into the amniotic fluids. The placentas were retained. The fetuses (approx 100 and 150 days of age) were decomposed; M agalactiae subsp bovis was recovered from several tissues of the fetuses, the placentas, and fetal fluids. The same organism was given by intraperitoneal injection to 2 other pregnant (130 and 180 days, respectively) cows. At necropsy of the latter 36 days later, placentitis was severe; M agalactiae subsp bovis was recovered from the placentas of both cows and from the fetus of 1 cow. Control cows given sterile mycoplasma cultural medium by intraamnion or intraperitoneal injection did not abort and were not infected. When first recovered from the bovine placenta and fetus, M agalactiae subsp bovis grew slowly in liquid medium and assumed bizarre colonial morphology on solidified medium. Colonies were small (0.1 to 0.5 mm) and dark and lacked halos, but they reacted specifically in the direct fluorescent antibody test with equine M agalactiae subsp bovis antiserum. After 1 or 2 subcultures, the isolates grew at a normal rate and displayed their usual colonial morphology.

  1. Experimental arthritis induced by a clinical Mycoplasma fermentans isolate

    PubMed Central

    Rivera, Antonio; Yáñez, Antonio; León-Tello, Gloria; Gil, Constantino; Giono, Silvia; Barba, Eduardo; Cedillo, Lilia

    2002-01-01

    Background Mycoplasma fermentans has been associated with rheumatoid arthritis. Recently, it was detected in the joints and blood of patients with rheumatoid arthritis, but it is not clear yet how the bacteria enter the body and reach the joints. The purpose of this study was to determine the ability of M. fermentans to induce experimental arthritis in rabbits following inoculation of the bacteria in the trachea and knee joints. Methods P-140 and PG-18 strains were each injected in the knee joints of 14 rabbits in order to evaluate and compare their arthritogenicity. P-140 was also injected in the trachea of 14 rabbits in order to test the ability of the bacteria to reach the joints and induce arthritis. Results M. fermentans produced an acute arthritis in rabbits. Joint swelling appeared first in rabbits injected with P-140, which caused a more severe arthritis than PG-18. Both strains were able to migrate to the uninoculated knee joints and they were detected viable in the joints all along the duration of the experiment. Changes in the synovial tissue were more severe by the end of the experiment and characterized by the infiltration of neutrophils and substitution of adipose tissue by connective tissue. Rabbits intracheally injected with P-140 showed induced arthritis and the bacteria could be isolated from lungs, blood, heart, kidney, spleen, brain and joints. Conclusion M. fermentans induced arthritis regardless of the inoculation route. These findings may help explain why mycoplasmas are commonly isolated from the joints of rheumatic patients. PMID:12057023

  2. A Major Determinant for Gliding Motility in Mycoplasma genitalium

    PubMed Central

    Martinelli, Luca; Lalli, Daniela; García-Morales, Luis; Ratera, Mercè; Querol, Enrique; Piñol, Jaume; Fita, Ignacio; Calisto, Bárbara M.

    2015-01-01

    Several mycoplasmas, such as the emergent human pathogen Mycoplasma genitalium, developed a complex polar structure, known as the terminal organelle (TO), responsible for a new type of cellular motility, which is involved in a variety of cell functions: cell division, adherence to host cells, and virulence. The TO cytoskeleton is organized as a multisubunit dynamic motor, including three main ultrastructures: the terminal button, the electrodense core, and the wheel complex. Here, we describe the interaction between MG200 and MG491, two of the main components of the TO wheel complex that connects the TO with the cell body and the cell membrane. The interaction between MG200 and MG491 has a KD in the 80 nm range, as determined by surface plasmon resonance. The interface between the two partners was confined to the “enriched in aromatic and glycine residues” (EAGR) box of MG200, previously described as a protein-protein interaction domain, and to a 25-residue-long peptide from the C-terminal region of MG491 by surface plasmon resonance and NMR spectroscopy studies. An atomic description of the MG200 EAGR box binding surface was also provided by solution NMR. An M. genitalium mutant lacking the MG491 segment corresponding to the peptide reveals specific alterations in cell motility and cell morphology indicating that the MG200-MG491 interaction plays a key role in the stability and functioning of the TO. PMID:25471372

  3. Molecular characterization of the uncultivatable hemotropic bacterium Mycoplasma haemofelis.

    PubMed

    Barker, Emily N; Darby, Alistair C; Helps, Chris R; Peters, Iain R; Heesom, Kate J; Arthur, Christopher J; Crossett, Ben; Hughes, Margaret A; Radford, Alan D; Tasker, Séverine

    2011-07-12

    Mycoplasma haemofelis is a pathogenic feline hemoplasma. Despite its importance, little is known about its metabolic pathways or mechanism of pathogenicity due to it being uncultivatable. The recently sequenced M. haemofelis str. Langford 1 genome was analysed and compared to those of other available hemoplasma genomes.Analysis showed that in hemoplasmas genes involved in carbohydrate metabolism are limited to enzymes of the glycolytic pathway, with glucose appearing to be the sole energy source. The majority of the pentose phosphate pathway enzymes that catalyze the de novo synthesis of ribonucleotides were absent, as were cell division protein FtsZ and chaperonins GroEL/ES. Uncharacterized protein paralogs containing putative surface expression motifs, comprised 62% of M. haemofelis and 19% of Mycoplasma suis genome coverage respectively, the majority of which were present in a small number of unstructured islands. Limited mass spectrometry and immunoblot data matched a number of characterized proteins and uncharacterized paralogs, confirming their expression and immunogenicity in vivo.These data have allowed further characterization of these important pathogens, including their limited metabolic capabilities, which may contribute to their uncultivatable status. A number of immunogenic proteins, and a potential mechanism for host immune system evasion, have been identified.

  4. Molecular characterization of the uncultivatable hemotropic bacterium Mycoplasma haemofelis

    PubMed Central

    2011-01-01

    Mycoplasma haemofelis is a pathogenic feline hemoplasma. Despite its importance, little is known about its metabolic pathways or mechanism of pathogenicity due to it being uncultivatable. The recently sequenced M. haemofelis str. Langford 1 genome was analysed and compared to those of other available hemoplasma genomes. Analysis showed that in hemoplasmas genes involved in carbohydrate metabolism are limited to enzymes of the glycolytic pathway, with glucose appearing to be the sole energy source. The majority of the pentose phosphate pathway enzymes that catalyze the de novo synthesis of ribonucleotides were absent, as were cell division protein FtsZ and chaperonins GroEL/ES. Uncharacterized protein paralogs containing putative surface expression motifs, comprised 62% of M. haemofelis and 19% of Mycoplasma suis genome coverage respectively, the majority of which were present in a small number of unstructured islands. Limited mass spectrometry and immunoblot data matched a number of characterized proteins and uncharacterized paralogs, confirming their expression and immunogenicity in vivo. These data have allowed further characterization of these important pathogens, including their limited metabolic capabilities, which may contribute to their uncultivatable status. A number of immunogenic proteins, and a potential mechanism for host immune system evasion, have been identified. PMID:21749699

  5. Molecular biology of mycoplasmas: from the minimum cell concept to the artificial cell.

    PubMed

    Cordova, Caio M M; Hoeltgebaum, Daniela L; Machado, Laís D P N; Santos, Larissa Dos

    2016-01-01

    Mycoplasmas are a large group of bacteria, sorted into different genera in the Mollicutes class, whose main characteristic in common, besides the small genome, is the absence of cell wall. They are considered cellular and molecular biology study models. We present an updated review of the molecular biology of these model microorganisms and the development of replicative vectors for the transformation of mycoplasmas. Synthetic biology studies inspired by these pioneering works became possible and won the attention of the mainstream media. For the first time, an artificial genome was synthesized (a minimal genome produced from consensus sequences obtained from mycoplasmas). For the first time, a functional artificial cell has been constructed by introducing a genome completely synthesized within a cell envelope of a mycoplasma obtained by transformation techniques. Therefore, this article offers an updated insight to the state of the art of these peculiar organisms' molecular biology.

  6. Detection of Mycoplasma Contamination Directly from Culture Supernatant Using Polymerase Chain Reaction.

    PubMed

    Pisal, R V; Hrebíková, H; Chvátalová, J; Kunke, D; Filip, S; Mokrý, J

    2016-01-01

    Ensuring mycoplasma-free cell culture is of prime importance as they severely affect cellular characteristics leading to experimental artefacts and spurious results. Various methods persist for mycoplasma detection; out of the whole array of methods polymerase chain reaction (PCR) is the most favoured one because it is highly sensitive, specific and quick. The PCR-based detection procedure involves three steps: cell culture supernatant collection, DNA isolation, and PCR. We have modified this procedure so that cell culture supernatant can directly be used for PCR without the need for DNA extraction. This modification makes the procedure quicker and more sensitive because loss of mycoplasma DNA is prevented and this loss becomes more significant when the level of mycoplasma contamination is very low.

  7. A disseminated Mycoplasma hominis infection in a patient with an underlying defect in humoral immunity.

    PubMed

    Nulens, Eric; Van Praet, Jens; Selleslag, Dominik; Van Landschoot, Thomas; Dekeyzer, Dieter; Descheemaecker, Patrick; Reynders, Marijke

    2016-06-01

    Non-urogenital Mycoplasma hominis infections are rare, but may cause life-threatening complications. We describe a case of disseminated M. hominis infection with extensive abscess formation in an immunocompromised patient with iatrogenic hypogammaglobulinemia under rituximab treatment.

  8. The infection of Mycoplasma hominis after total knee replacement: Case report and literature review.

    PubMed

    Qiu, Hong-Jiu; Lu, Wei-Ping; Li, Min; Wang, Zi-Ming; Du, Quan-Yin; Wang, Ai-Min; Xiong, Yan

    2017-08-01

    The Mycoplasma hominis infection is a rare postoperative complication after joint replacement. Based on our knowledge, there were only two cases reported by Korea all over the world currently. A case of postoperative Mycoplasma hominis infection after total knee replacement in our hospital was reported in this article. It was confirmed through mass spectrometer and Mycoplasma cultivation and treated by the first stage debridement, polyethylene insert replacement, and then drainage and irrigation combined with sensitive antibiotics after the operation. We observed that the C reactive protein (CRP) level correlates with the development of disease, while the erythrocyte sedimentation rate (ESR) remains at a high level, indicating the relevance between the Mycoplasma hominis infection caused by knee joint replacement and CRP. This study aims to report the case and review relevant literature. Copyright © 2017. Production and hosting by Elsevier B.V.

  9. [Effects of the symbiosis of Trichomonas vaginalis with Mycoplasma hominis on ferredoxin gene].

    PubMed

    Liu, Xiaodong; Wen, Wenjing; Xue, Changgui

    2011-08-01

    We isolated 30 Trichomonas vaginalis for the PCR detection from the gynecological outpatients in the Affiliated Hospital of Zhengzhou University using the specific 16s rDNA primers of Mycoplasma hominis. The results showed that there were 25 cases of Mycoplasma hominis infection, with the infection rate of 83.33%. This gave a clew that the symbiosis of Trichomonas vaginalis with Mycoplasma hominis may be of certain generality in China. We sequenced the ferredoxin gene of 10 Trichomonas vaginalis where 5 Mycoplasma hominis were positive and five negative, and found that the ferredoxin (Fd) gene of the 10 Trichomonas vaginalis were exactly the same. But compared to the genes in the GenBank, a comparative analysis of the gene revealed that there were 3 more ctg bases at the 200th position of encoding leucine, but this did not lead to changes in reading frame. The gene homology was 99%.

  10. Treatment of resistant mycoplasma infection in immunocompromised patients with a new pleuromutilin antibiotic.

    PubMed

    Heilmann, C; Jensen, L; Jensen, J S; Lundstrom, K; Windsor, D; Windsor, H; Webster, D

    2001-11-01

    Patients with primary antibody deficiency (PAD) are prone to mycoplasma infection with unusual strains which may be resistant to conventional antibiotics. Mycoplasmas were isolated from the joint fluid (Ureaplasma urealyticum) of two PAD patients with arthritis and from the cerebral spinal fluid (Mycoplasma maculosum) in one with meningitis, the latter probably originating from the patient's dog. Combinations of doxycycline and quinolones or macrolides failed to clear the infections, but after demonstrating in-vitro sensitivity to the pleuromutilin, Econor, for two of the isolates, all three patients responded to oral treatment with Econor. The infection was completely eradicated in two patients, with the emergence of a resistant strain in the third. Mycoplasma infection should be considered in PAD patients with unexplained sepsis. Pleuromutilins such as Econor are powerful new anti-mycoplasmal agents which provide an additional therapeutic option when patients fail to respond to conventional antibiotics. Copyright 2001 The British Infection Society.

  11. Mycoplasma hyopneumoniae and Mycoplasma flocculare differential domains from orthologous surface proteins induce distinct cellular immune responses in mice.

    PubMed

    Leal, Fernanda Munhoz Dos Anjos; Virginio, Veridiana Gomes; Martello, Carolina Lumertz; Paes, Jéssica Andrade; Borges, Thiago J; Jaeger, Natália; Bonorino, Cristina; Ferreira, Henrique Bunselmeyer

    2016-07-15

    Mycoplasma hyopneumoniae and Mycoplasma flocculare are two genetically close species found in the swine respiratory tract. Despite their similarities, while M. hyopneumoniae is the causative agent of porcine enzootic pneumonia, M. flocculare is a commensal bacterium. Genomic and transcriptional comparative analyses so far failed to explain the difference in pathogenicity between these two species. We then hypothesized that such difference might be, at least in part, explained by amino acid sequence and immunological or functional differences between ortholog surface proteins. In line with that, it was verified that approximately 85% of the ortholog surface proteins from M. hyopneumoniae 7448 and M. flocculare present one or more differential domains. To experimentally assess possible immunological implications of this kind of difference, the extracellular differential domains from one pair of orthologous surface proteins (MHP7448_0612, from M. hyopneumoniae, and MF_00357, from M. flocculare) were expressed in E. coli and used to immunize mice. The recombinant polypeptides (rMHP61267-169 and rMF35767-196, respectively) induced distinct cellular immune responses. While, rMHP61267-169 induced both Th1 and Th2 responses, rMF35767-196 induced just an early pro-inflammatory response. These results indicate that immunological properties determined by differential domains in orthologous surface protein might play a role in pathogenicity, contributing to elicit specific and differential immune responses against each species.

  12. [Case of tuberculous pleurisy distinguished from pleurisy caused by Mycoplasma infection].

    PubMed

    Nakamura, Kazuyoshi; Yamanaka, Tohru; Yoshioka, Yuichi; Horio, Yuko; Iyama, Shinji; Suzumura, Tomoko; Esaki, Toshihiro

    2013-04-01

    We report a case of tuberculous pleurisy that required differentiation from pleurisy caused by Mycoplasma infection. A 28-year-old woman presented to a clinic with fever and pain on the left side of her chest. A chest radiograph revealed pleural effusion in the left thorax, and the condition was diagnosed as bacterial pleurisy. The patient was referred to our hospital because of an increase in the pleural effusion despite antibiotic treatment. Mycoplasma infection was suspected because the patient was young, the white blood cell count was not elevated, and the result of the ImmunoCard Mycoplasma test (IC) for Mycoplasma pneumoniae-specific IgM antibodies was positive. However, the fever persisted even after treatment with azithromycin and pazufloxacin. The left pleural effusion was exudative, with lymphocytosis and high adenosine deaminase (ADA) levels. The results of the QuantiFERON test were positive. Therefore, tuberculous pleurisy was diagnosed, and the effusion subsided after treatment with standard anti-tuberculosis chemotherapy. Although detection of Mycoplasma infection using the IC is rapid and simple, the accuracy of this test is poor. The patient was first diagnosed with pleurisy of Mycoplasma origin because of a single high-particle agglutination titer of 1: 320 and because of the presence of exudative pleural effusion with lymphocytosis and elevated ADA levels, which has been reported in patients with Mycoplasma infection. The results of the IC test and the ADA level of the pleural effusion might not be reliable when distinguishing between tuberculous pleurisy and pleurisy caused by Mycoplasma infection.

  13. Purification of Encephalitozoon Cultures Contaminated by Mycoplasmas by Murine Intraperitoneal Inoculation

    PubMed Central

    Ridoux, Olivier; Foucault, Cédric; Drancourt, Michel

    1998-01-01

    Encephalitozoon species are strict intracellular microsporidia. Cocultures with eukaryotic cell lines can become accidently contaminated by mycoplasmas. We propose a decontamination protocol based on differential cell targeting after intraperitoneal inoculation in mice. Mycoplasma-free microsporidia were isolated from the brains and spleens of inoculated mice 24 h postinoculation by using the centrifugation shell vial system. Identification was confirmed by direct sequencing of PCR-amplified 16S rRNA. PMID:9666031

  14. Cross reactivity among the swine mycoplasmas as identified by protein microarray.

    PubMed

    Petersen, Andrew C; Oneal, David C; Seibel, Janice R; Poel, Kylie; Daum, Courtney L; Djordjevic, Steven P; Minion, F Chris

    2016-08-30

    Mycoplasmas are cell wall-less bacteria that infect a variety of animals in a species-specific manner. In swine, Mycoplasma hyopneumoniae is the most virulent and presents the most disease and economic problems to the swine industry. Serological assays are commonly used to assess colonization and disease, but antigenic cross-reactivity between M. hyopneumoniae and other mycoplasma species, most notably Mycoplasma hyorhinis, Mycoplasma hyosynoviae and Mycoplasma flocculare, is a concern. The extent of cross-reactivity has not been thoroughly investigated. These studies were designed to identify M. hyopneumoniae proteins that are recognized by rabbit hyperimmune sera raised against the other swine mycoplasmas. Our results indicate extensive cross-reactivity between M. flocculare and M. hyopneumoniae, which explains previous reports seen with ELISA assays. Only three of the thirty-nine M. hyopneumoniae proteins tested showed no cross reactivity with the other three swine mycoplasmas, mhp182 (42kDa C-terminal fragment), mhp638 and mhp684 (C-terminal fragment). Two proteins, mhp384 and mhp511, were cross-reactive with hyperimmune sera generated against three of the four species. None of the anti-M. hyorhinis hyperimmune sera reacted to any of the M. hyopneumoniae proteins. These results suggest that inapparent M. flocculare infections could produce positive responses in M. hyopneumoniae serological assays due to cross-reactivity, and that M. hyosynoviae infections are less likely to do so and M. hyorhinis infections unlikely to affect assay results. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Suitability of peracetic acid for sterilization of media for mycoplasma cultures.

    PubMed Central

    Wutzler, P; Sprössig, M; Peterseim, H

    1975-01-01

    The utility of peracetic acid for sterilization of serum and yeast extract additions to mycoplasma medium was studied by culturing six Mycoplasma species. Culture media containing additions that had been sterilized with peracetic acid proved to be as good as filtered components. The use of 0.05 to 0.1% peracetic acid is recommended to sterilize the serum and yeast extract additions since savings in time and equipment can be accomplished. PMID:1100656

  16. Effects of mycoplasma contamination on phenotypic expression of mitochondrial mutants in human cells

    SciTech Connect

    Doersen, C.J.; Stanbridge, E.J.

    1981-04-01

    HeLa cells sensitive to the mitochondrial protein synthesis inhibitors erythromycin (ERY) and chloramphenicol (CAP) and HeLa variants resistant to the effects of these drugs were purposefully infected with drug-sensitive and -resistnat mycoplasma strains. Mycoplasma hyorhinis and the ERY-resistant strain of Mycoplasma orale, MO-ERY/sup r/, did not influence the growth of HeLa and ERY-resistant ERY2301 cells in the presence or absence of ERY. M. hyorhinis also did not affect the growth of HeLa and CAP-resistant Cap-2 cells in the presence or absence of CAP. However, both HeLa and Cap-2 cells infected with the CAP-resistant strain of M. hyorhinis, MH-CAP/sup r/, were more sensitive to the cytotoxix effect of CAP. This maybe due to the glucose dependence of the cells, which was compromised by the increased utilization of glucose by MH-CAP/sup r/ in these infected cell cultures. In vitro protein synthesis by isolated mitochondria was significantly altered by mycoplasma infection of the various cell lines. A substantial number of mycoplasmas copurified with the mitochondria, resulting in up to a sevenfold increase in the incorporation of (/sup 3/H)leucine into the trichloroacetic acid-insoluble material. More importantly, the apparent drug sensitivity or resistance of mitchondrial preparations from mycoplasma-infected cells reflected the drug sensitivity or resistance of the contaminating mycoplasmas. These results illustrate the hazards in interpreting mitochondrial protein synthesis data derived from mycoplasma-infected cell lines, particularly putative mitochondrially encoded mutants resistant to inhibitors of mitochondrial protein synthesis.

  17. Cranial Sixth-Nerve Palsy and Eosinophilia in an Outbreak of Mycoplasma Pneumonia

    PubMed Central

    Wing, Howard J.; Lindzon, Martin

    1987-01-01

    The authors discuss a case in which three siblings presented with Mycoplasma pneumonia. All three had a typical rise in complement fixation antibody titres. However, the sibling with the highest titre also developed cranial sixth-nerve palsy; in addition, she was the only one of the three who did not have an eosinophilia. The authors review the symptomatology of Mycoplasma pneumonia and the involvement of the central nervous system. PMID:21263943

  18. MRI appearances of the CNS manifestations of Mycoplasma pneumoniae: a report of two cases.

    PubMed

    Francis, D A; Brown, A; Miller, D H; Wiles, C M; Bennett, E D; Leigh, N

    1988-09-01

    Two patients are reported with Mycoplasma pneumoniae-related cervical myelitis. Magnetic resonance imaging in each case demonstrated clinically silent lesions suggesting more extensive neurological involvement. This supports the concept of widespread immunologically mediated disease occurring as a remote effect of initial M. pneumoniae respiratory infection. Differences from the MRI appearances of a patient with mycoplasma-related Guillian-Barré syndrome imply that more than one antigenic determinant is involved.

  19. The detection of "Candidatus Mycoplasma haemobos" in cattle and buffalo in China.

    PubMed

    Su, Q L; Song, H Q; Lin, R Q; Yuan, Z G; Yang, J F; Zhao, G H; Huang, W Y; Zhu, Xing Quan

    2010-12-01

    "Candidatus Mycoplasma haemobos" is a hemoplasma species found in cattle and has been recently reported in Switzerland and Japan. In this study, "Candidatus Mycoplasma haemobos" was shown to occur in cattle and buffalo in tropical China by PCR amplification and sequence analysis of the 16S rRNA gene from blood samples. Based on the 16S rDNA sequence, a specific PCR assay was developed. Occurrence of "Candidatus Mycoplasma haemobos" in cattle and buffalo in Guangxi, China, was determined by examining 25 buffalo blood samples, 12 yellow cattle blood samples and 42 dairy cow blood samples. The results showed that 32% (8/25) of buffalo, 41.7% (5/12) of yellow cattle, and 14.3% (6/42) of dairy cows were positive for "Candidatus Mycoplasma haemobos", respectively. Direct sequencing of representative PCR products confirmed that the amplified partial 16S rDNA sequence represented "Candidatus Mycoplasma haemobos". This is the first report of "Candidatus Mycoplasma haemobos" in buffalo, yellow cattle, and dairy cows in China.

  20. Integrative Conjugative Elements Are Widespread in Field Isolates of Mycoplasma Species Pathogenic for Ruminants

    PubMed Central

    Mick, Virginie; Dordet-Frisoni, Emilie; Marenda, Marc Serge; Sirand-Pugnet, Pascal; Blanchard, Alain

    2014-01-01

    Comparative genomics have revealed massive horizontal gene transfer (HGT) between Mycoplasma species sharing common ruminant hosts. Further results pointed toward an integrative conjugative element (ICE) as an important contributor of HGT in the small-ruminant-pathogen Mycoplasma agalactiae. To estimate the prevalence of ICEs in ruminant mycoplasmas, we surveyed their occurrence in a collection of 166 field strains representing 4 (sub)species that are recognized as major pathogens. Based on available sequenced genomes, we first defined the conserved, minimal ICE backbone as composed of 4 coding sequences (CDSs) that are evenly distributed and predicted to be essential for ICE chromosomal integration-excision and horizontal transfer. Screening of the strain collection revealed that these 4 CDSs are well represented in ruminant Mycoplasma species, suggesting widespread occurrence of ICEs. Yet their prevalence varies within and among species, with no correlation found with the individual strain history. Extrachromosomal ICE forms were also often detected, suggesting that ICEs are able to circularize in all species, a first and essential step in ICE horizontal transfer. Examination of the junction of the circular forms and comparative sequence analysis of conserved CDSs clearly pointed toward two types of ICE, the hominis and spiroplasma types, most likely differing in their mechanism of excision-integration. Overall, our data indicate the occurrence and maintenance of functional ICEs in a large number of field isolates of ruminant mycoplasmas. These may contribute to genome plasticity and gene exchanges and, presumably, to the emergence of diverse genotypes within pathogenic mycoplasmas of veterinary importance. PMID:25527550

  1. Adenovirus and mycoplasma infection in an ornate box turtle (Terrapene ornata ornata) in Hungary.

    PubMed

    Farkas, Szilvia L; Gál, János

    2009-07-02

    A female, adult ornate box turtle (Terrapene ornata ornata) with fatty liver was submitted for virologic examination in Hungary. Signs of an adenovirus infection including degeneration of the liver cells, enlarged nuclei and intranuclear inclusion bodies were detected by light microscopic examination. The presence of an adenovirus was later confirmed by obtaining partial sequence data from the adenoviral DNA-dependent DNA-polymerase. Phylogenetic analyses revealed that this novel chelonian adenovirus was distinct from previously described reptilian adenoviruses, not belonging to any of the recognized genera of the family Adenoviridae. As a part of the routine diagnostic procedure for chelonians the detection of herpes-, rana- and iridoviruses together with Mycoplasma spp. was attempted. Amplicons were generated by a general mycoplasma polymerase chain reaction (PCR) targeting the 16S/23S ribosomal RNA (rRNA) intergenic spacer region, as well as, a specific Mycoplasma agassizii PCR targeting the 16S rRNA gene. Based on the analyses of partial sequences of the 16S rRNA gene, the Mycoplasma sp. of the ornate box turtle seemed to be identical with the recently described eastern box turtle (Terrapene carolina carolina) Mycoplasma sp. This is the first report of a novel chelonian adenovirus and a mycoplasma infection in an ornate box turtle (T. ornata ornata) in Europe.

  2. Integrative conjugative elements are widespread in field isolates of Mycoplasma species pathogenic for ruminants.

    PubMed

    Tardy, Florence; Mick, Virginie; Dordet-Frisoni, Emilie; Marenda, Marc Serge; Sirand-Pugnet, Pascal; Blanchard, Alain; Citti, Christine

    2015-03-01

    Comparative genomics have revealed massive horizontal gene transfer (HGT) between Mycoplasma species sharing common ruminant hosts. Further results pointed toward an integrative conjugative element (ICE) as an important contributor of HGT in the small-ruminant-pathogen Mycoplasma agalactiae. To estimate the prevalence of ICEs in ruminant mycoplasmas, we surveyed their occurrence in a collection of 166 field strains representing 4 (sub)species that are recognized as major pathogens. Based on available sequenced genomes, we first defined the conserved, minimal ICE backbone as composed of 4 coding sequences (CDSs) that are evenly distributed and predicted to be essential for ICE chromosomal integration-excision and horizontal transfer. Screening of the strain collection revealed that these 4 CDSs are well represented in ruminant Mycoplasma species, suggesting widespread occurrence of ICEs. Yet their prevalence varies within and among species, with no correlation found with the individual strain history. Extrachromosomal ICE forms were also often detected, suggesting that ICEs are able to circularize in all species, a first and essential step in ICE horizontal transfer. Examination of the junction of the circular forms and comparative sequence analysis of conserved CDSs clearly pointed toward two types of ICE, the hominis and spiroplasma types, most likely differing in their mechanism of excision-integration. Overall, our data indicate the occurrence and maintenance of functional ICEs in a large number of field isolates of ruminant mycoplasmas. These may contribute to genome plasticity and gene exchanges and, presumably, to the emergence of diverse genotypes within pathogenic mycoplasmas of veterinary importance.

  3. Identification of Chlamydiae and Mycoplasma species in ruminants with ocular infections.

    PubMed

    Gupta, S; Chahota, R; Bhardwaj, B; Malik, P; Verma, S; Sharma, M

    2015-02-01

    Infectious keratoconjunctivitis (IKC) is a highly contagious ocular inflammatory condition, which is often reported in domestic small and large ruminants. Multiple infectious aetiologies are reported to be involved, but information about the role of certain fastidious bacterial pathogens such as chlamydiae and mycoplasmas is limited in India. Hence, this study was performed to determine the role of these pathogens and their identification by molecular approach. A total of 53 samples from 31 ovine, 14 caprine and eight bovine having clinical symptoms were collected and tested using species-specific PCR tests for chlamydiae and mycoplasmas followed by nucleotide sequence analysis. The results showed 77.41, 14.29 and 25% samples were chlamydiae positive in ovine, caprine and bovine, respectively, whereas 41.93, 14.29 and 37.5% prevalence of mycoplasma infection was detected in ovine, caprine and bovines, respectively. Chlamydophila abortus, Chlamydophila psittaci, Mycoplasma arginini and Mycoplasma hyorhinis were detected from tested samples. To the best of our knowledge, this is the first time these species are identified in IKC cases from India. Coinfection of both chlamydial and mycoplasmal species was detected in eight IKC cases of ovine which suggest synergistic roles played by both chlamydiae and mycoplasma in IKC samples.

  4. Prevalence of Ureaplasma urealyticum and Mycoplasma hominis infection in unselected infertile men.

    PubMed

    Salmeri, Mario; Valenti, Daniela; La Vignera, Sandro; Bellanca, Salvatore; Morello, Angela; Toscano, M Antonietta; Mastrojeni, Silvana; Calogero, Aldo E

    2012-04-01

    In this study, we investigated the prevalence of Ureaplasma urealyticum and Mycoplasma hominis infection among 250 unselected infertile men, the presence of urogenital symptoms in infected men and the effects of these microorganisms on the conventional sperm parameters. Urethral samples were obtained using a swab inserted 3-4 cm into the urethral meatus. Ureaplasma urealyticum and Mycoplasma hominis were detected by the kit Mycofast R evolution 3 Elitech Microbiology (Elitech Microbiology, Signes, France). Ureaplasma urealyticum was detected in 15.6% of the cases and Mycoplasma hominis in 3.6%. One patients had a co-infection with both pathogens. About 41% of the infertile patients with mycoplasma infection had urogenital symptoms. A lower number of patients with mycoplasma infection had normal sperm parameters compared with non-infected infertile men, but this frequency showed only a trend compared to non-infected patients (Chi-square=3.61; P=0.057), and a significantly higher percentage of patients with oligo-astheno-teratozoospermia (Chi-square=127.3; P<0.0001), or asthenozoospermia alone (Chi-square=5.74; P<0.05) compared to non-infected infertile patients. In conclusion, this study showed an elevated prevalence of Ureaplasma urealyticum and Mycoplasma hominis infection in unselected men attending an infertility outpatient clinic and that the presence of these microorganisms is associated with a higher percentage of patients with abnormal sperm parameters.

  5. Frequency and antimicrobial sensitivity of Ureaplasma urealyticum and Mycoplasma hominis in patients with vaginal discharge.

    PubMed

    Díaz, Leonor; Cabrera, Luis E; Fernández, Tania; Ibáñez, Inailay; Torres, Yulian; Obregón, Yakelí; Rivero, Yanelys

    2013-10-01

    Determination of antimicrobial sensitivity helps establish adequate treatment and avoids future genital tract diseases in women of fertile age. In Cuba, prevalence of mycoplasma in patients with vaginal discharge is unknown. The objective of this research was to determine frequency and antimicrobial sensitivity of Ureaplasma urealyticum and Mycoplasma hominis in women with vaginal discharge through analysis of laboratory data from vaginal smears from 255 patients referred to the Municipal Hygiene and Epidemiology Center in Güines, Mayabeque Province, Cuba. Mycoplasma System Plus (Italy) was used for detection, identification, count and sensitivity testing. The finding of mycoplasmas in almost two thirds of specimens examined suggests that the sexually active female population should be screened for these bacteria and that barrier contraception methods should be promoted to decrease their spread and prevent longterm sequelae. Such updating of local patterns of antimicrobial resistance supports decision making for best treatment options in patients with these infections. Our results should help clinicians in our area choose an antibiotic, and also confirm the utility of Mycoplasma System Plus for mycoplasma research in resource-scarce settings, to benefit individual and population health.

  6. Epidemiology of Ureaplasma urealyticum and Mycoplasma hominis in the semen of male outpatients with reproductive disorders.

    PubMed

    Zhu, Xiaofei; Li, Min; Cao, Huiling; Yang, Xuewen; Zhang, Chunbing

    2016-08-01

    The aim of the present study was to investigate the association between Mycoplasma infection and infertility in male outpatients among a Chinese population. Epidemiological data, including prevalence, age distribution and antibiotic resistance profile of patients with an Ureaplasma urealyticum or Mycoplasma hominis infection were collected between 2009 and 2012. Among the 7,374 individuals analyzed, 3,225 patients (43.7%) were determined to be positive for infection with U. urealyticum, M. hominis or for both Mycoplasmas. Among the positive cultures, U. urealyticum was detected most frequently, while M. hominis was rarely found. The age range of 25-34 years was the preferred period for the positive detection. Tetracyclines and josamycin were the most effective agents against both genital Mycoplasmas, including in the case of co-infection. Macrolides (erythromycin, roxithromycin, azithromycin, clarithromycin except for josamycin) were effective against the majority of U. urealyticum clinical isolates, but were naturally resisted by M. hominis in this study. Fluoroquinolones had the lowest activity against U. urealyticum, particularly in cases of M. hominis co-infection. Furthermore, fluoroquinolones showed a similar pattern of drug resistance against M. hominis to that of U. urealyticum. Antibiotic resistance did not vary significantly over the test period. Notably, an elevated multi-drug resistance rate was observed in patients co-infected with both Mycoplasmas. In light of the epidemiological characteristics of genital Mycoplasmas in male infertility patients, the present results may aid Chinese clinicians to implement rational drug usage and avoid the overuse of antibiotics.

  7. Epidemiology of Ureaplasma urealyticum and Mycoplasma hominis in the semen of male outpatients with reproductive disorders

    PubMed Central

    Zhu, Xiaofei; Li, Min; Cao, Huiling; Yang, Xuewen; Zhang, Chunbing

    2016-01-01

    The aim of the present study was to investigate the association between Mycoplasma infection and infertility in male outpatients among a Chinese population. Epidemiological data, including prevalence, age distribution and antibiotic resistance profile of patients with an Ureaplasma urealyticum or Mycoplasma hominis infection were collected between 2009 and 2012. Among the 7,374 individuals analyzed, 3,225 patients (43.7%) were determined to be positive for infection with U. urealyticum, M. hominis or for both Mycoplasmas. Among the positive cultures, U. urealyticum was detected most frequently, while M. hominis was rarely found. The age range of 25–34 years was the preferred period for the positive detection. Tetracyclines and josamycin were the most effective agents against both genital Mycoplasmas, including in the case of co-infection. Macrolides (erythromycin, roxithromycin, azithromycin, clarithromycin except for josamycin) were effective against the majority of U. urealyticum clinical isolates, but were naturally resisted by M. hominis in this study. Fluoroquinolones had the lowest activity against U. urealyticum, particularly in cases of M. hominis co-infection. Furthermore, fluoroquinolones showed a similar pattern of drug resistance against M. hominis to that of U. urealyticum. Antibiotic resistance did not vary significantly over the test period. Notably, an elevated multi-drug resistance rate was observed in patients co-infected with both Mycoplasmas. In light of the epidemiological characteristics of genital Mycoplasmas in male infertility patients, the present results may aid Chinese clinicians to implement rational drug usage and avoid the overuse of antibiotics. PMID:27443698

  8. Degradation of nuclease-stabilized RNA oligonucleotides in Mycoplasma-contaminated cell culture media.

    PubMed

    Hernandez, Frank J; Stockdale, Katie R; Huang, Lingyan; Horswill, Alexander R; Behlke, Mark A; McNamara, James O

    2012-02-01

    Artificial RNA reagents such as small interfering RNAs (siRNAs) and aptamers often must be chemically modified for optimal effectiveness in environments that include ribonucleases. Mycoplasmas are common bacterial contaminants of mammalian cell cultures that are known to produce ribonucleases. Here we describe the rapid degradation of nuclease-stabilized RNA oligonucleotides in a human embryonic kidney 293 (HEK) cell culture contaminated with Mycoplasma fermentans, a common species of mycoplasma. RNA with 2'-fluoro- or 2'-O-methyl- modified pyrimidines was readily degraded in conditioned media from this culture, but was stable in conditioned media from uncontaminated HEK cells. RNA completely modified with 2'-O-methyls was not degraded in the mycoplasma-contaminated media. RNA zymogram analysis of conditioned culture media and material centrifuged from the media revealed several distinct protein bands (ranging from 30 to 68 kDa) capable of degrading RNA with 2'-fluoro- or 2'-O-methyl-modified pyrimidines. Finally, the mycoplasma-associated nuclease was detected in material centrifuged from the contaminated culture supernatants in as little as 15 minutes with an RNA oligo-containing 2'-O-methyl-modified pyrimidines and labeled with a 5'-fluorescein amidite (FAM) and 3'-quencher. These results suggest that mycoplasma contamination may be a critical confounding variable for cell culture experiments involving RNA-based reagents, with particular relevance for applications involving naked RNA (e.g., aptamer-siRNA chimeras).

  9. Investigations into the seasonal presence of Mycoplasma species in fattening lambs.

    PubMed

    Fernández, Sara; Galapero, Javier; Rey, Joaquín; Pérez, Carlos Javier; Ramos, Alfonso; Rosales, Rubén; Ayling, Roger; Alonso, Juan Manuel; Gómez, Luis

    2016-06-01

    The presence of infection with Mycoplasma species in association with lung consolidation, environmental temperature and relative humidity was investigated in 410 clinically healthy fattening lambs from five different feedlots in Extremadura (southwestern Spain). Isolates of Mycoplasma species were obtained (n= 117), including Mycoplasma ovipneumoniae (n = 18) and Mycoplasma arginini (n = 99). Two seasonal periods were identified. The first period, which included February, March, September, October, and November, had an average temperature of 17.5 ± 4.7 °C and a relative humidity of 61.3 ± 15.8%. The second seasonal period, which included the months from April to August, had an average temperature of 22.9 ± 5.5 °C and a relative humidity of 48.4 ± 10.7%. Most Mycoplasma species were isolated from the second seasonal period, indicating that higher temperatures and lower relative humidity favour the presence of Mycoplasma species. M. arginini was also associated with lung consolidation.

  10. Standardized methods and quality control limits for agar and broth microdilution susceptibility testing of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum.

    PubMed

    Waites, Ken B; Duffy, Lynn B; Bébéar, Cécile M; Matlow, Anne; Talkington, Deborah F; Kenny, George E; Totten, Patricia A; Bade, Donald J; Zheng, Xiaotian; Davidson, Maureen K; Shortridge, Virginia D; Watts, Jeffrey L; Brown, Steven D

    2012-11-01

    An international multilaboratory collaborative study was conducted to develop standard media and consensus methods for the performance and quality control of antimicrobial susceptibility testing of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum using broth microdilution and agar dilution techniques. A reference strain from the American Type Culture Collection was designated for each species, which was to be used for quality control purposes. Repeat testing of replicate samples of each reference strain by participating laboratories utilizing both methods and different lots of media enabled a 3- to 4-dilution MIC range to be established for drugs in several different classes, including tetracyclines, macrolides, ketolides, lincosamides, and fluoroquinolones. This represents the first multilaboratory collaboration to standardize susceptibility testing methods and to designate quality control parameters to ensure accurate and reliable assay results for mycoplasmas and ureaplasmas that infect humans.

  11. High quality draft genomes of the Mycoplasma mycoides subsp. mycoides challenge strains Afadé and B237.

    PubMed

    Fischer, Anne; Santana-Cruz, Ivette; Hegerman, Jan; Gourlé, Hadrien; Schieck, Elise; Lambert, Mathieu; Nadendla, Suvarna; Wesonga, Hezron; Miller, Rachel A; Vashee, Sanjay; Weber, Johann; Meens, Jochen; Frey, Joachim; Jores, Joerg

    2015-01-01

    Members of the Mycoplasma mycoides cluster' represent important livestock pathogens worldwide. Mycoplasma mycoides subsp. mycoides is the etiologic agent of contagious bovine pleuropneumonia (CBPP), which is still endemic in many parts of Africa. We report the genome sequences and annotation of two frequently used challenge strains of Mycoplasma mycoides subsp. mycoides, Afadé and B237. The information provided will enable downstream 'omics' applications such as proteomics, transcriptomics and reverse vaccinology approaches. Despite the absence of Mycoplasma pneumoniae like cyto-adhesion encoding genes, the two strains showed the presence of protrusions. This phenotype is likely encoded by another set of genes.

  12. Molecular detection of Mycoplasma haemofelis and 'Candidatus Mycoplasma haemominutum' Infection in cats by direct PCR using whole blood without DNA extraction.

    PubMed

    Watanabe, Masashi; Hisasue, Masaharu; Souma, Takehisa; Ohshiro, Shuichi; Yamada, Takatsugu; Tsuchiya, Ryo

    2008-10-01

    Detection of hemotropic Mycoplasma spp. infection was attempted in cats by PCR using whole blood without DNA extraction. A total 46 of 54 (85%) cats with suspected Mycoplasma spp. infection showed a positive reaction, corresponding completely with the results of standard PCR testing. The direct PCR assay was sensitive enough to detect more than 0.0061% parasitemia for ;C. M. haemominutum' and 0.0075% parasitemia for M. haemofelis. These data indicate that the direct PCR assay might be sufficient for use as a tool in clinical examinations.

  13. MLVA typing of Mycoplasma hyopneumoniae bacterins and field strains

    PubMed Central

    Tamiozzo, P.; Zamora, R.; Lucchesi, P. M. A.; Estanguet, A.; Parada, J.; Carranza, A.; Camacho, P.; Ambrogi, A.

    2015-01-01

    Because of the lack of information about both the genetic characteristics of Mycoplasma hyopneumoniae commercial vaccines and their relationship with field strains, the authors attempted to identify genetic subtypes of some M hyopneumoniae bacterins, and to compare them with M. hyopneumoniae field strains. Six commercial M hyopneumoniae bacterins and 28 bronchoalveolar lavages from pigs at slaughter from three herds were analysed by Multiple-Locus Variable number tandem repeat Analysis (MLVA) on p146R1, p146R3, H4, H5 and p95 loci. The results obtained showed the presence of more than one M hyopneumoniae genotype in some pigs and also in one of the bacterins analysed. It is also worth noting that MLVA typing allowed the distinction among circulating field strains and also when comparing them with vaccine strains, which, knowing the relatedness among them, could be useful in the research of the efficacy of the vaccines. PMID:26495127

  14. Unitary step of gliding machinery in Mycoplasma mobile.

    PubMed

    Kinosita, Yoshiaki; Nakane, Daisuke; Sugawa, Mitsuhiro; Masaike, Tomoko; Mizutani, Kana; Miyata, Makoto; Nishizaka, Takayuki

    2014-06-10

    Among the bacteria that glide on substrate surfaces, Mycoplasma mobile is one of the fastest, exhibiting smooth movement with a speed of 2.0-4.5 μm⋅s(-1) with a cycle of attachment to and detachment from sialylated oligosaccharides. To study the gliding mechanism at the molecular level, we applied an assay with a fluorescently labeled and membrane-permeabilized ghost model, and investigated the motility by high precision colocalization microscopy. Under conditions designed to reduce the number of motor interactions on a randomly oriented substrate, ghosts took unitary 70-nm steps in the direction of gliding. Although it remains possible that the stepping behavior is produced by multiple interactions, our data suggest that these steps are produced by a unitary gliding machine that need not move between sites arranged on a cytoskeletal lattice.

  15. Mycoplasmas hyorhinis in different regions of cuba. diagnosis

    PubMed Central

    Lobo, Evelyn; Poveda, Carlos; Gupta, Rakesh; Suarez, Alejandro; Hernández, Yenney; Ramírez, Ana; Poveda, José B.

    2011-01-01

    M. hyorhinis is considered one of the etiological agents of arthritis in sucking pigs, but recently as seen, some strains can produce pneumonia that could not be distinguished from the mycoplasmosis caused by M. hyopneumoniae. The study was conducted to research the presence of Mycoplasma hyorhinis (M. hyorhinis ) in different regions of the country from exudates of pig lungs with typical EP lesions. Exudates from 280 pig lungs with typical EP lesions were studied using molecular techniques such as PCR, real time PCR and amplification of the 16S-23S rRNA. It was detected that the 66% of the samples studied resulted positive to M. hyorhinis, and the presence of this species was detected in all the provinces. Amplification and studies on the intergenic region 16S-23S of M. hyorhinis rRNA demonstrated the existing variability among strains of a same species. This study is the first report on M. hyorhinis detection in Cuba. PMID:24031686

  16. Mycoplasma genitalium: Is It a Sexually Transmitted Pathogen?

    PubMed

    Manhart, Lisa E; Kay, Noa

    2010-07-01

    Mycoplasma genitalium is an emerging pathogen that has been detected in the male and female reproductive tracts. It is an established cause of nongonococcal urethritis and evidence linking it to cervicitis, endometritis, and tubal factor infertility is accumulating. Whether a pathogen is sexually transmitted has important implications for clinical management because partner management strategies are an essential part of the treatment plan for sexually transmitted infections. However, mere detection in the genital tract and associations with reproductive tract disease are insufficient to conclude that an organism is sexually transmitted. Therefore, to assess whether M. genitalium is sexually transmitted, we evaluated the literature in terms of associations with established risk factors for other sexually transmitted infections, comparisons of sexually experienced individuals to nonsexually experienced individuals, consideration of other modes of transmission, assessment of concordant infection status among sexual partners, and examination of molecular strain typing in concordantly infected partners.

  17. Identification of Haemobartonella felis (Mycoplasma haemofelis) in captive nondomestic cats.

    PubMed

    Haefner, Monika; Burke, Thomas J; Kitchell, Barbara E; Lamont, Leigh A; Schaeffer, David J; Behr, Melissa; Messick, Joanne B

    2003-06-01

    This study was undertaken to determine whether Haemobartonella felis (Mycoplasma haemofelis), the causative bacterial agent of feline infectious anemia, infects nondomestic cats. Routine complete blood count and polymerase chain reaction (PCR) were performed to detect the gene for 16S ribosomal RNA for the organism. Sixty-four blood samples were collected from 54 nondomestic cats, including tigers (Panthera tigris), cheetahs (Acinonyx jubatus), lions (P. leo), mountain lions (Felis concolor), snow leopards (P. unica), and a jaguar (P. onca). Some cats were sampled on two or three different dates. Two tigers were positive for H. felis by PCR analysis. As previously described in domestic cats, the parasitemia appears to be intermittent in nondomestic cats.

  18. Mycoplasma bovis: Mechanisms of Resistance and Trends in Antimicrobial Susceptibility

    PubMed Central

    Lysnyansky, Inna; Ayling, Roger D.

    2016-01-01

    Mycoplasma bovis is a cell-wall-less bacterium and belongs to the class Mollicutes. It is the most important etiological agent of bovine mycoplasmoses in North America and Europe, causing respiratory disease, mastitis, otitis media, arthritis, and reproductive disease. Clinical disease associated with M. bovis is often chronic, debilitating, and poorly responsive to antimicrobial therapy, resulting in significant economic loss, the full extent of which is difficult to estimate. Until M. bovis vaccines are universally available, sanitary control measures and antimicrobial treatment are the only approaches that can be used in attempts to control M. bovis infections. However, in vitro studies show that many of the current M. bovis isolates circulating in Europe have high minimum inhibitory concentrations (MIC) for many of the commercially available antimicrobials. In this review we summarize the current MIC trends indicating the development of antimicrobial resistance in M. bovis as well as the known molecular mechanisms by which resistance is acquired. PMID:27199926

  19. Cold hemagglutinin cross-reactivity with Mycoplasma pneumoniae.

    PubMed Central

    Janney, F A; Lee, L T; Howe, C

    1978-01-01

    Convalescent sera from proven cases of infection with Mycoplasma pneumoniae, and rabbit antisera to M. pneumoniae and to human erythrocyte glycoprotein contained cold hemagglutinins which were reactive only for human erythrocytes. Only the human serum cold agglutinins were inhibited by soluble integral glycoproteins derived from human erythrocyte ghosts by treatment with chloroform-methanol. Rabbit antiserum to chloroform-methanol glycoprotein, as well as to M. pneumoniae, fixed complement with either M. pneumoniae or chloroform-methanol glycoprotein antigens. The findings support the hypothesis that the cold agglutinins elicited by M. pneumoniae infection represent a cross-reaction between determinants common to erythrocyte glycoprotein containing I antigen and the membrane of M. pneumoniae. PMID:83298

  20. A field study of Mycoplasma bovis infection in cattle.

    PubMed

    Feenstra, A; Bisgaard Madsen, E; Friis, N F; Meyling, A; Ahrens, P

    1991-05-01

    After an outbreak of mastitis in cattle caused by Mycoplasma bovis a study was made in 5 herds with recent cases (principal herds) and in 4 control herds. In the principal herds, M. bovis was isolated from milk samples, nasal swabs, and from one vaginal swab. M. bovis was also isolated from nasal swabs of calves in 2 of the 4 control herds, whereas all milk samples and vaginal swabs from the control herds were negative. Evaluation of serum antibody titres to M. bovis among non-mastitic animals of 3 principal herds and 1 control herd showed no difference in distribution of the titre values, which generally were low. However, cows excreting M. bovis in the milk had high antibody titres. The way of introduction to the herds and the spread of the infection within the herds could not be established by the study, which was supplemented by a DNA restriction fragment analysis of a number of M. bovis isolates.