Sample records for mycoplasma hominis strains

  1. Complete Genome Sequence of Mycoplasma hominis Strain Sprott (ATCC 33131), Isolated from a Patient with Nongonococcal Urethritis

    PubMed Central

    Foecking, Mark F.

    2015-01-01

    Presented here is the complete and annotated genome sequence of Mycoplasma hominis Sprott (ATCC 33131). The chromosome comprises 695,214 bp, which is approximately 30 kb larger than the syntenic genome of M. hominis PG21T. Tetracycline resistance of strain Sprott is most probably conferred by the tetM determinant, harbored on a mosaic transposon-like structure. PMID:26159538

  2. Detection and prevention of mycoplasma hominis infection

    DOEpatents

    DelVecchio, Vito G. (Scranton, PA); Gallia, Gary L. (Philadelphia, PA); McCleskey, Ferne K. (San Antonio, TX)

    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.

  3. Arginine metabolism in Trichomonas vaginalis infected with Mycoplasma hominis

    PubMed Central

    Morada, Mary; Manzur, Mafruha; Lam, Brian; Tan, Cho; Tachezy, Jan; Rappelli, Paola; Dessì, Daniele; Fiori, Pier L.; Yarlett, Nigel

    2010-01-01

    Both Mycoplasma hominis and Trichomonas vaginalis utilize arginine as an energy source via the arginine dihydrolase (ADH) pathway. It has been previously demonstrated that M. hominis forms a stable intracellular relationship with T. vaginalis; hence, in this study we examined the interaction of two localized ADH pathways by comparing T. vaginalis strain SS22 with the laboratory-generated T. vaginalis strain SS22-MOZ2 infected with M. hominis MOZ2. The presence of M. hominis resulted in an approximately 16-fold increase in intracellular ornithine and a threefold increase in putrescine, compared with control T. vaginalis cultures. No change in the activity of enzymes of the ADH pathway could be demonstrated in SS22-MOZ2 compared with the parent SS22, and the increased production of ornithine could be attributed to the presence of M. hominis. Using metabolic flow analysis it was determined that the elasticity of enzymes of the ADH pathway in SS22-MOZ2 was unchanged compared with the parent SS22; however, the elasticity of ornithine decarboxylase (ODC) in SS22 was small, and it was doubled in SS22-MOZ2 cells. The potential benefit of this relationship to both T. vaginalis and M. hominis is discussed. PMID:20656780

  4. Pharmacokinetics of Moxifloxacin in an Infant with Mycoplasma hominis Meningitis

    PubMed Central

    Watt, Kevin M; Massaro, Matthew M; Smith, Brian; Cohen-Wolkowiez, Michael; Benjamin, Daniel K.; Laughon, Matthew M

    2012-01-01

    Treatment of Mycoplasma hominis meningitis in infants is limited by a lack of consensus regarding therapy and limited pharmacokinetic data for agents to which M. hominis is susceptible. We report the successful treatment of a premature infant with M. hominis meningitis with doxycycline and moxifloxacin and provide a pharmacokinetic profile of moxifloxacin. PMID:22016080

  5. In vitro susceptibilities of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum to sparfloxacin and PD 127391.

    PubMed

    Waites, K B; Duffy, L B; Schmid, T; Crabb, D; Pate, M S; Cassell, G H

    1991-06-01

    The in vitro activities of two investigational quinolones, sparfloxacin (previously designated AT 4140) and PD 127391, were determined for 30 strains each of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum and compared with those of ciprofloxacin, tetracycline, clindamycin, and erythromycin. Erythromycin was the most active compound against M. pneumoniae (maximum MIC, less than 0.008 microgram/ml). PD 127391 (MICs, less than 0.008 to 0.031 microgram/ml), sparfloxacin (MICs, less than 0.008 to 0.25 microgram/ml), clindamycin (MICs, less than 0.008 to 0.5 microgram/ml), and tetracycline (MICs, 0.063 to 0.25 microgram/ml) were superior to ciprofloxacin (MICs, 0.5 to 2 microgram/ml). Sparfloxacin and PD 127391 were active against M. hominis (MICs, less than 0.008 to 0.031 microgram/ml for each) at concentrations comparable to those of clindamycin (MICs, less than 0.008 to 0.063 microgram/ml) and at concentrations lower than those of ciprofloxacin (MICs, 0.125 to 0.5 microgram/ml). As expected, M. hominis was resistant to erythromycin (MICs, 32 to greater than or equal to 256 micrograms/ml). For U. urealyticum, PD 127391 (MICs, 0.031 to 0.5 microgram/ml) and sparfloxacin (MICs, 0.063 to 1 microgram/ml) were superior to erythromycin (MICs, 0.25 to 4 micrograms/ml), ciprofloxacin (MICs, 0.5 to 8 micrograms/ml), and clindamycin (MICs, 0.25 to 64 micrograms/ml. Both new quinolones were equally active against tetracycline-susceptible as well as resistant strains of M. hominis and U. urealyticum. The possible influence of medium components and/or pH on MICs was evaluated by testing a Staphylococcus aureus reference strain with each antibiotic in SP-4 broth and 10-B broth and comparing the results with published MICs for this strain. MICs determined in 10-B broth for erythromycin were affected most. This study shows that the activities of sparfloxacin and PD 127391 are similar to one another and comparable or superior to those of other drugs used to treat mycoplasmal infections. The MICs of both new quinolones were consistently 2 to several dilutions lower than those of ciprofloxacin for each species. PMID:1929260

  6. Host and pathogen interaction during vaginal infection by Trichomonas vaginalis and Mycoplasma hominis or Ureaplasma urealyticum

    Microsoft Academic Search

    Cindy van der Schee; Hans J. F Sluiters; Willem I van der Meijden; Patricia van Beek; Paul Peerbooms; Henri Verbrugh; Alex van Belkum

    2001-01-01

    Vaginal infections by Trichomonas vaginalis and Mycoplasma hominis have been shown to be associated. Since M. hominis and Ureaplasma urealyticum are similar pathogens, both belonging to the class of the mycoplasmata, we describe here a molecular study into the interdependence of U. urealyticum and T. vaginalis during infection. Susceptibility towards infection by U. urealyticum depends on genetic polymorphism in the

  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 this pestering pathogen. PMID:23326599

  8. Evaluation of the Etest for susceptibility testing of Mycoplasma hominis and Ureaplasma urealyticum.

    PubMed

    Dósa, E; Nagy, E; Falk, W; Szöke, I; Ballies, U

    1999-04-01

    The Etest was used for antibiotic susceptibility testing of Mycoplasma hominis and Ureaplasma urealyticum isolates and the results were compared with those obtained with the broth microdilution method. For 50 clinical isolates of M. hominis the MICs of doxycycline, ofloxacin and ciprofloxacin agreed within +/- one dilution and +/- two dilutions in 82-98% and 98-100% of cases, respectively. The MICs of erythromycin, azithromycin, doxycycline, ofloxacin and ciprofloxacin were evaluated for 50 clinical isolates of U. urealyticum. The corresponding levels of agreement were 70-98% and 94-100%, respectively. Reference isolates M. hominis PG-21 and U. urealyticum T-960 were also used. The Etest seems to be an alternative method for determination of MICs of antibiotics with M. hominis and U. urealyticum. PMID:10350390

  9. A case report of Mycoplasma hominis brain abscess identified by MALDI-TOF mass spectrometry.

    PubMed

    Pailhoriès, H; Rabier, V; Eveillard, M; Mahaza, C; Joly-Guillou, M-L; Chennebault, J-M; Kempf, M; Lemarié, C

    2014-12-01

    We report the case of a 43-year-old man with a Mycoplasma hominis brain abscess occurring after a cranial trauma, which was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The presence of colonies on classic blood agar plates and the use of MALDI-TOF MS, a valuable diagnostic tool that identified M. hominis due to its presence in the VITEK MS database, allowed the rapid diagnosis of this infection. PMID:25449252

  10. Isolation and Molecular Identification of Mycoplasma Hominis in Infertile Female and Male Reproductive System

    PubMed Central

    Jamalizadeh Bahaabadi, Samaneh; Mohseni Moghadam, Naeime; Kheirkhah, Babak; Farsinejad, Alireza; Habibzadeh, Victoria

    2014-01-01

    Background: Infection of urogenital system with Mycoplasma potentially affect reproductive system and increases infants mortalities. Therefore, detection of these organisms is an important issue that should be considered and appropriate diagnostic methods should be used to identify these microorganisms. In the female reproductive system, infection can affect different parts of the cervix, endometrium, and fallopian tube. The extent of this infection in different diseases and its pathogenesis might be related to anatomic site of involvement. Some infections can lead to infertility in both males and females. Genital infection with Mycoplasmas have devastating effects on reproductive organs and cause fertility disorders and mortality in infants. In recent years, many studies have been conducted to isolate these pathogens; however, the isolates have not been identified so far. Objectives: The aim of this study was to determine the molecular identity of Mycoplasma hominis isolated from infertile female and male reproductive system in the Infertility Center of Kerman. Materials and Methods: This descriptive study was performed purposefully on 100 infertile females and 100 infertile males who were referred to the Infertility Center of Kerman during a six-month period. The collected samples of semen and vaginal swabs were examined for the presence of M. hominis by PCR. The samples with positive results in PCR were selected for molecular identification. Alignment of samples sequence was performed using MEGA 5 software through Neighbor-joining method. Results: Among 100 samples from infertile males, the presence of genus Mycoplasma was confirmed in 45 cases of which 15 cases were infected with M. hominis. Among 100 samples from infertile female, the presence of genus Mycoplasma was confirmed in 43 cases of which 18 case were infected with M. hominis. The positive samples were sequenced and the phylogenetic tree was plotted. Conclusions: The results showed that 37.5% of infertile males and females were infected with M. hominis. Analysis of the nucleotide sequences of the study isolates indicates a particular variety among these isolates. In comparing the isolates in the study, a very little genotypic similarity was found among some of them. PMID:25738116

  11. Mycoplasma hominis ssp. associated endocarditis with myocardial necrosis in an alpaca (Vicugna pacos) in Manitoba in 2011.

    PubMed

    Tomczyk, Krzysztof M; Copeland, Shelagh; Postey, Rosemary; Ngeleka, Musangu

    2015-02-01

    Severe endocarditis with myonecrosis, moderate to severe pleural and pericardial effusions, and mild ascites were found on necropsy in 3 alpacas. Mycoplasma hominis ssp. was detected on polymerase chain reaction (PCR) of fresh affected endocardial tissue in 1 alpaca. PMID:25694661

  12. Draft Genome Sequence of Lactobacillus hominis Strain CRBIP 24.179T, Isolated from Human Intestine

    E-print Network

    Paris-Sud XI, Université de

    Draft Genome Sequence of Lactobacillus hominis Strain CRBIP 24.179T, Isolated from Human Intestine genome sequence of the strain Lactobacillus hominis CRBIP 24.179T, isolated from a human clinical sample, Clermont D, Loux V, Bizet C, Bouchier C. 2013. Draft genome sequence of Lactobacillus hominis strain CRBIP

  13. Long-Term Survival and Intracellular Replication of Mycoplasma hominis in Trichomonas vaginalis Cells: Potential Role of the Protozoon in Transmitting Bacterial Infection

    Microsoft Academic Search

    D. Dessi; Giuseppe Delogu; Eleonora Emonte; Maria Rosaria Catania; P. L. Fiori; P. Rappelli

    2005-01-01

    The existence of a symbiotic relationship between Trichomonas vaginalis and Mycoplasma hominis, which is the first reported example of symbiosis between two obligate human pathogens, has been recently reported by our research group. In this work, we examined the cellular location of M. hominis in respect to T. vaginalis .B y using gentamicin protection assays, double immunofluorescence, and confocal microscopy,

  14. Isolation of Mycoplasma hominis in critically ill patients with pulmonary infections: clinical and microbiological analysis in an intensive care unit

    Microsoft Academic Search

    Celia García; Estibaliz Ugalde; Idoia Monteagudo; Ana Saez; Jesús Agüero; Luis Martinez-Martinez; Eduardo Miñambres

    2007-01-01

    Objective  \\u000a Mycoplasma hominis is a well recognized extragenital pathogen. However, it is an uncommon cause of respiratory infections in critically ill\\u000a patients admitted to the intensive care unit (ICU).\\u000a \\u000a \\u000a \\u000a Design and setting  Prospective clinical investigation in a 21-bed ICU in a university hospital.\\u000a \\u000a \\u000a \\u000a Patients  Seven patients requiring intensive care who developed a ICU-acquired pneumonia in which M. hominis was recovered from bronchoalveolar lavage and pleural fluid

  15. Differential vaginal expression of interleukin-1 system cytokines in the presence of Mycoplasma hominis and Ureaplasma urealyticum in pregnant women.

    PubMed Central

    Doh, Kunihiko; Barton, Parrin T; Korneeva, Irina; Perni, Sriram C; Bongiovanni, Ann Marie; Tuttle, Sara L; Skupski, Daniel W; Witkin, Steven S

    2004-01-01

    OBJECTIVE: The genital mycoplasmas, Ureaplasma urealyticum and Mycoplasma hominis, are commonly identified in the vagina of healthy pregnant women. However, these microorganisms are the most common isolates from the amniotic fluids of women in preterm labor. The mechanisms responsible for vaginal colonization and ascent to the uterus remain undetermined. We evaluated the association between U. urealyticum and M. hominis vaginal colonization and the presence of pro-inflammatory and anti-inflammatory interleukin-1 system components in asymptomatic pregnant women of different ethnicities. METHODS: Vaginal specimens, obtained from 224 first trimester pregnant women, were assayed for interleukin-1beta (IL-1beta) and IL-1 receptor antagonist (IL-1ra) concentrations by ELISA. U. urealyticum and M. hominis vaginal colonization were identified by polymerase chain reaction (PCR). RESULTS: Vaginal colonization with M. hominis was identified in 37 (16.5%) women, and was more prevalent in black (18.9%) and Hispanic (20.9%) than in white (4.2%) women (p = 0.01). U. urealyticum was present in 84 (37.5%) women and there was no ethnic disparity in its detection. M. hominis colonization was associated with elevated median vaginal IL-1beta concentrations in both black women (p = 0.02) and Hispanic women (p = 0.04), and was unrelated to vaginal IL-1ra concentrations. In marked contrast, U. urealyticum colonization was associated with elevations in vaginal IL-1ra levels, but not with IL-1beta concentrations, in black women (p = 0.02) and Hispanic women (p < 0.0001) and marginally in white women (p = 0.06). CONCLUSION: M. hominis colonization in healthy pregnant women is associated with localized pro-inflammatory immune activation, while U. urealyticum colonization is associated with immune suppression. PMID:15739821

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

    Microsoft Academic Search

    A. T. R. Vasconcelos; H. B. Ferreira; C. V. Bizarro; S. L. Bonatto; M. O. Carvalho; P. M. Pinto; D. F. Almeida; L. G. P. Almeida; R. Almeida; L. Alves-Filho; E. N. Assuncao; V. A. C. Azevedo; M. R. Bogo; M. M. Brigido; M. Brocchi; H. A. Burity; A. A. Camargo; S. S. Camargo; M. S. Carepo; D. M. Carraro; J. C. de Mattos Cascardo; L. A. Castro; G. Cavalcanti; G. Chemale; R. G. Collevatti; C. W. Cunha; B. Dallagiovanna; B. P. Dambros; Odir A. Dellagostin; C. Falcao; F. Fantinatti-Garboggini; M. S. S. Felipe; L. Fiorentin; G. R. Franco; N. S. A. Freitas; D. Frias; T. B. Grangeiro; E. C. Grisard; C. T. Guimaraes; M. Hungria; S. N. Jardim; M. A. Krieger; J. P. Laurino; L. F. A. Lima; M. I. Lopes; E. L. S. Loreto; H. M. F. Madeira; G. P. Manfio; A. Q. Maranhao; C. T. Martinkovics; S. R. B. Medeiros; M. A. M. Moreira; M. Neiva; C. E. Ramalho-Neto; M. F. Nicolas; S. C. Oliveira; R. F. C. Paixao; F. O. Pedrosa; S. D. J. Pena; M. Pereira; L. Pereira-Ferrari; I. Piffer; L. S. Pinto; D. P. Potrich; A. C. M. Salim; F. R. Santos; R. Schmitt; M. P. C. Schneider; A. Schrank; I. S. Schrank; A. F. Schuck; H. N. Seuanez; D. W. Silva; R. Silva; S. C. Silva; C. M. A. Soares; K. R. L. Souza; R. C. Souza; C. C. Staats; M. B. R. Steffens; S. M. R. Teixeira; T. P. Urmenyi; M. H. Vainstein; L. W. Zuccherato; A. J. G. Simpson; A. Zaha

    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

  17. [Frequency of detection of Ureaplasma urealyticum and Mycoplasma hominis in cervical canal and the Douglas pouch of infertile and fertile women].

    PubMed

    Grze?ko, Joanna; Elias, Marek; Maczy?ska, Beata; Kasprzykowska, Urszula; T?acza?a, Magdalena; Goluda, Marian

    2007-01-01

    The group of organisms commonly referred to as genital mycoplasmas comprise species most often found in genitourinary tract of sexually active adults as common commensal inhabitants, or pathogens which can possibly cause many different pathologies like: non-gonococcal urethritis, bacterial vaginosis, cervicitis, endometritis or pelvic inflammatory disease. The problem of their morbidity and the possible influence they have on human fertility is still not clear. The aim of this study was to find out whether two investigated species- Ureaplasma urealyticum and Mycoplasma hominis can be detect more often in a group of infertile women. 74 women participated in the study and were assigned to one of 2 groups of patients: infertile women and fertile women without any sign of genital tract infection. Swabs from the cervical canal of the uterus and the fluid from the Douglas pouch were taken during the gynecological examination and laparoscopic procedure. Two diagnostic methods were used: biochemical method- commercial diagnostic kit- Mycoplasma IST 2 and PCR method. The results showed that Ureaplasma urealyticum and Mycoplasma hominis were detected among both fertile and infertile women with nearly the same frequency, much more often in cervical canal than in the Douglas pouch. Ureaplasma urealyticum was more common pathogen than Mycoplasma hominis in both groups and locations. The achieved results point out that the role of genital mycoplasmas in human infertility is still unclear and require further investigations. PMID:17929414

  18. Genome Sequence of Mycoplasma hyorhinis Strain DBS 1050

    PubMed Central

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

    2014-01-01

    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. PMID:24604646

  19. Genome Sequence of Mycoplasma hyorhinis Strain DBS 1050.

    PubMed

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

    2014-01-01

    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. PMID:24604646

  20. Complete Genome Sequence of Mycoplasma synoviae Strain WVU 1853T.

    PubMed

    May, Meghan A; Kutish, Gerald F; Barbet, Anthony F; Michaels, Dina L; Brown, Daniel R

    2015-01-01

    A hybrid sequence assembly of the complete Mycoplasma synoviae type strain WVU 1853(T) genome was compared to that of strain MS53. The findings support prior conclusions about M. synoviae, based on the genome of that otherwise uncharacterized field strain, and provide the first evidence of epigenetic modifications in M. synoviae. PMID:26021934

  1. Complete Genome Sequence of Mycoplasma synoviae Strain WVU 1853T

    PubMed Central

    Kutish, Gerald F.; Barbet, Anthony F.; Michaels, Dina L.

    2015-01-01

    A hybrid sequence assembly of the complete Mycoplasma synoviae type strain WVU 1853T genome was compared to that of strain MS53. The findings support prior conclusions about M. synoviae, based on the genome of that otherwise uncharacterized field strain, and provide the first evidence of epigenetic modifications in M. synoviae. PMID:26021934

  2. The Complete Genome Sequence of Mycoplasma bovis Strain Hubei-1

    Microsoft Academic Search

    Yuan Li; Huajun Zheng; Yang Liu; Yanwei Jiang; Jiuqing Xin; Wei Chen; Zhiqiang Song; Herman Tse

    2011-01-01

    Infection by Mycoplasma bovis (M. bovis) can induce diseases, such as pneumonia and otitis media in young calves and mastitis and arthritis in older animals. Here, we report the finished and annotated genome sequence of M. bovis strain Hubei-1, a strain isolated in 2008 that caused calf pneumonia on a Chinese farm. The genome of M. bovis strain Hubei-1 contains

  3. Genome Sequence of Mycoplasma capricolum subsp. capripneumoniae Strain M1601

    PubMed Central

    Chu, Yuefeng; Gao, Pengchen; Zhao, Ping; He, Ying; Liao, Nancy; Jackman, Shaun; Zhao, Yongjun; Birol, Inanc; Duan, Xiaobo; Lu, Zhongxin

    2011-01-01

    Mycoplasma capricolum subsp. capripneumoniae is the causative agent of contagious caprine pleuropneumonia, a devastating disease of goats listed by the World Organization for Animal Health. Here we report the first complete genome sequence of this organism (strain M1601, a clinically isolated strain from China). PMID:21994928

  4. Phylogenetic Analysis of Mycoplasma Strain ISM1499 and Its Assignment to the Acholeplasma oculi Strain Cluster

    Microsoft Academic Search

    SERGEY ARTIUSHIN; MEL DUVALL; F. CHRIS MINION

    1995-01-01

    A mycoplasma strain designated ISM1499 was used to develop a mycoplasma genetic system (G. G. Mahairas and F. C. Minion, J. Bacteriol. 171:1775-1780,1989; G. G. Mahairas, C. Jian, and F. C. Minion, Gene 93:61-65, 1990), but phenotypic inconsistencies led to the conclusion that this organism had been classified incorrectly as a member of the species Mycoplasma pulmonis. Studies were initiated

  5. Highly specific and efficient primers for in-house multiplex PCR detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis and Ureaplasma urealyticum

    PubMed Central

    2014-01-01

    Background Although sophisticated methodologies are available, the use of endpoint polymerase chain reaction (PCR) to detect 16S rDNA genes remains a good approach for estimating the incidence and prevalence of specific infections and for monitoring infections. Considering the importance of the early diagnosis of sexually transmitted infections (STIs), the development of a sensitive and affordable method for identifying pathogens in clinical samples is needed. Highly specific and efficient primers for a multiplex polymerase chain reaction (m-PCR) system were designed in silico to detect the 16S rDNA genes of four bacteria that cause genital infections, and the PCR method was developed. Methods The Genosensor Probe Designer (GPD) (version 1.0a) software was initially used to design highly specific and efficient primers for in-house m-PCR. Single-locus PCR reactions were performed and standardised, and then primers for each locus in turn were added individually in subsequent amplifications until m-PCR was achieved. Amplicons of the expected size were obtained from each of the four bacterial gene fragments. Finally, the analytical specificity and limits of detection were tested. Results Because they did not amplify any product from non-STI tested species, the primers were specific. The detection limits for the Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis and Ureaplasma urealyticum primer sets were 5.12?×?105, 3.9?×?103, 61.19?×?106 and 6.37?×?105 copies of a DNA template, respectively. Conclusions The methodology designed and standardised here could be applied satisfactorily for the simultaneous or individual detection of Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma hominis and Ureaplasma urealyticum. This method is at least as efficient as other previously described methods; however, this method is more affordable for low-income countries. PMID:24997675

  6. The Complete Genome Sequence of Mycoplasma bovis Strain Hubei-1

    PubMed Central

    Li, Yuan; Zheng, Huajun; Liu, Yang; Jiang, Yanwei; Xin, Jiuqing; Chen, Wei; Song, Zhiqiang

    2011-01-01

    Infection by Mycoplasma bovis (M. bovis) can induce diseases, such as pneumonia and otitis media in young calves and mastitis and arthritis in older animals. Here, we report the finished and annotated genome sequence of M. bovis strain Hubei-1, a strain isolated in 2008 that caused calf pneumonia on a Chinese farm. The genome of M. bovis strain Hubei-1 contains a single circular chromosome of 953,114 bp with a 29.37% GC content. We identified 803 open reading frames (ORFs) that occupy 89.5% of the genome. While 34 ORFs were Hubei-1 specific, 662 ORFs had orthologs in the M. bovis type strain PG45 genome. Genome analysis validated lateral gene transfer between M. bovis and the Mycoplasma mycoides subspecies mycoides, while phylogenetic analysis found that the closest M. bovis neighbor is Mycoplasma agalactiae. Glycerol may be the main carbon and energy source of M. bovis, and most of the biosynthesis pathways were incomplete. We report that 47 lipoproteins, 12 extracellular proteins and 18 transmembrane proteins are phase-variable and may help M. bovis escape the immune response. Besides lipoproteins and phase-variable proteins, genomic analysis found two possible pathogenicity islands, which consist of four genes and 11 genes each, and several other virulence factors including hemolysin, lipoate protein ligase, dihydrolipoamide dehydrogenase, extracellular cysteine protease and 5?-nucleotidase. PMID:21731639

  7. Pathogenicity of Mycoplasma lipofaciens strain ML64 for turkey embryos.

    PubMed

    Lierz, M; Deppenmeier, S; Gruber, A D; Brokat, S; Hafez, H M

    2007-10-01

    Mycoplasma lipofaciens strain ML64, isolated from an egg of a northern goshawk (Accipiter gentilis), has been found to be pathogenic for chicken embryos causing mortality during the first 2 weeks of incubation. The same strain was inoculated in turkey embryos to evaluate its pathogenicity and its ability to be transmitted laterally in the hatchery. The strain was found to be pathogenic for turkey embryos, causing a high mortality (88.9%) during late incubation as well as haemorrhages of the legs, dwarfing, curled toes and a severe, multifocal, purulent to necrotizing bronchopneumonia. In addition, lateral transmission between turkey poults hatched from infected eggs and poults from non-infected controls was observed in the incubator. PMID:17899463

  8. The role of Mycoplasma strain F38 in contagious caprine pleuropneumonia (CCCP) in Kenya.

    PubMed

    MacOwan, K J; Minette, J E

    1977-11-01

    The results of a serological and cultural study of experimental and field cases of contagious caprine pleuropneumonia were consistent with an aetiological role for mycoplasma strain F38. This mycoplasma was isolated from 57 acute cases in 46 outbreaks of CCPP and from 87 experimental contact cases. Clinical data from experimental contact cases were assessed for comparison with field cases. PMID:595269

  9. Complete Genome Sequence of Mycoplasma capricolum subsp. capripneumoniae Strain 9231-Abomsa.

    PubMed

    Dupuy, Virginie; Thiaucourt, François

    2014-01-01

    Mycoplasma capricolum subsp. capripneumoniae is the etiological agent of contagious caprine pleuropneumonia. We report here the complete and annotated genome sequence of M. capricolum subsp. capripneumoniae strain 9231-Abomsa. PMID:25323727

  10. Complete Genome Sequence of Mycoplasma capricolum subsp. capripneumoniae Strain 9231-Abomsa

    PubMed Central

    Thiaucourt, François

    2014-01-01

    Mycoplasma capricolum subsp. capripneumoniae is the etiological agent of contagious caprine pleuropneumonia. We report here the complete and annotated genome sequence of M. capricolum subsp. capripneumoniae strain 9231-Abomsa. PMID:25323727

  11. Pathogenicity of Mycoplasma lipofaciens strain ML64, isolated from an egg of a Northern Goshawk (Accipiter gentilis), for chicken embryos

    Microsoft Academic Search

    M. Lierz; R. Stark; S. Brokat; H. M. Hafez

    2007-01-01

    Some Mycoplasma species are well-known avian pathogens and are of importance in poultry breeder flocks due to their pathogenic potential for embryos. Mycoplasmas are regularly detected in birds of prey, and a strain of Mycoplasma lipofaciens that was isolated from an egg of a Northern Goshawk (Accipiter gentilis) was examined for its pathogenicity in specific pathogen free chicken embryos since

  12. Effects of vaccination with F-strain Mycoplasma gallisepticum on egg production and quality parameters of commercial layer hens previously vaccinated with 6/85-strain Mycoplasma gallisepticum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An experiment was conducted to determine the effect of overlaying (revaccinating) F strain Mycoplasma gallisepticum (MG) at 22 or 45 weeks of age on commercial leghorn hens previously vaccinated with 6/85 strain MG at 10 weeks of age. The treatment groups include unvaccinated hens (group 1), hens r...

  13. Comparative genomic analysis of seven Mycoplasma hyosynoviae strains

    PubMed Central

    Bumgardner, Eric A; Kittichotirat, Weerayuth; Bumgarner, Roger E; Lawrence, Paulraj K

    2015-01-01

    Infection with Mycoplasma hyosynoviae can result in debilitating arthritis in pigs, particularly those aged 10 weeks or older. Strategies for controlling this pathogen are becoming increasingly important due to the rise in the number of cases of arthritis that have been attributed to infection in recent years. In order to begin to develop interventions to prevent arthritis caused by M. hyosynoviae, more information regarding the specific proteins and potential virulence factors that its genome encodes was needed. However, the genome of this emerging swine pathogen had not been sequenced previously. In this report, we present a comparative analysis of the genomes of seven strains of M. hyosynoviae isolated from different locations in North America during the years 2010 to 2013. We identified several putative virulence factors that may contribute to the ability of this pathogen to adhere to host cells. Additionally, we discovered several prophage genes present within the genomes of three strains that show significant similarity to MAV1, a phage isolated from the related species, M. arthritidis. We also identified CRISPR-Cas and type III restriction and modification systems present in two strains that may contribute to their ability to defend against phage infection. PMID:25693846

  14. The complete genome sequence of the avian pathogen Mycoplasma gallisepticum strain Rlow

    Microsoft Academic Search

    Leka Papazisi; Timothy S. Gorton; Gerald Kutish; Philip F. Markham; Glenn F. Browning; Steven Swartzell; Anup Madan; Greg Mahairas; Steven J. Geary

    2003-01-01

    The complete genome of Mycoplasma gallisepticum strain Rlow has been sequenced. The genome is composed of 996422 bp with an overall G+C content of 31 mol%. It contains 742 putative coding DNA sequences (CDSs), representing a 91% coding density. Function has been assigned to 469 of the CDSs, while 150 encode conserved hypothetical proteins and 123 remain as unique hypothetical

  15. Genital Mycoplasma infections and their resistance phenotypes in an African setting.

    PubMed

    Kouegnigan Rerambiah, L; Ndong, J-C; Medzegue, S; Elisee-Ndam, M; Djoba Siawaya, J F

    2015-06-01

    We investigated the antimicrobial susceptibilities of mycoplasmas in Gabonese men and women. A total of 1,332 men and women were included in the study. Sperm, urine, ureteral or vaginal swabs were collected from the subjects. Mycoplasmas identification and antimicrobial susceptibility to azithromycin, clarithromycin, erythromycin, josamycin, pristinamycin, doxycycline, tetracycline, ofloxacin and ciprofloxacin were tested using the Mycoplasma IST 2 kit. 794 subjects were positive for Mycoplasma. Respectively, 1.6 % and 82.24 % of subjects were singly infected with M. hominis and Ureaplasma urealyticum and 15.87 % had a mixed infection. M. hominis isolates were resistant to erythromycin and had an intermediate (I) to resistant (R) profile to azithromycin and clarithromycin. 84.6 % of M. hominis strains were sensitive (S) to josamycin and pristinamycin. 30.8 % and 92.3 % of M. hominis strains were sensitive to tetracycline and doxycycline, respectively. 76.9 and 84.6 % of M. hominis isolates were sensitive to ciprofloxacin and ofloxacin, respectively. The sensitivity rates of U. urealyticum strains were 45.23 %, 47.7 %, 63.84 %, 90.8 % and 92 % for azithromycin, erythromycin, clarithromycin, pristinamycin and josamycin, respectively. U. urealyticum strains showed 62.2 % and 79.7 % sensitivity to tetracycline and doxycycline, respectively. The resistance rates to azithromycin, clarithromycin and erythromycin for samples with mixed infection were 72.8 %, 84.7 % and 85.6 %, respectively. Josamycin and pristinamycin were 81.5 % effective on samples with mixed infection. The sensitivity rates of samples with mixed infection to tetracycline, doxycycline, ciprofloxacin and ofloxacin were 32 %, 69.6 %, 8.9 % and 18.5 %, respectively. Sub-Saharan Africa needs to use antibiotics rationally, as falling to do so would compromise the management of infectious diseases. PMID:25630539

  16. Comparative genomic analyses of Mycoplasma hyopneumoniae pathogenic 168 strain and its high-passaged attenuated strain

    PubMed Central

    2013-01-01

    Background Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia (EP), a mild, chronic pneumonia of swine. Despite presenting with low direct mortality, EP is responsible for major economic losses in the pig industry. To identify the virulence-associated determinants of M. hyopneumoniae, we determined the whole genome sequence of M. hyopneumoniae strain 168 and its attenuated high-passage strain 168-L and carried out comparative genomic analyses. Results We performed the first comprehensive analysis of M. hyopneumoniae strain 168 and its attenuated strain and made a preliminary survey of coding sequences (CDSs) that may be related to virulence. The 168-L genome has a highly similar gene content and order to that of 168, but is 4,483 bp smaller because there are 60 insertions and 43 deletions in 168-L. Besides these indels, 227 single nucleotide variations (SNVs) were identified. We further investigated the variants that affected CDSs, and compared them to reported virulence determinants. Notably, almost all of the reported virulence determinants are included in these variants affected CDSs. In addition to variations previously described in mycoplasma adhesins (P97, P102, P146, P159, P216, and LppT), cell envelope proteins (P95), cell surface antigens (P36), secreted proteins and chaperone protein (DnaK), mutations in genes related to metabolism and growth may also contribute to the attenuated virulence in 168-L. Furthermore, many mutations were located in the previously described repeat motif, which may be of primary importance for virulence. Conclusions We studied the virulence attenuation mechanism of M. hyopneumoniae by comparative genomic analysis of virulent strain 168 and its attenuated high-passage strain 168-L. Our findings provide a preliminary survey of CDSs that may be related to virulence. While these include reported virulence-related genes, other novel virulence determinants were also detected. This new information will form the foundation of future investigations into the pathogenesis of M. hyopneumoniae and facilitate the design of new vaccines. PMID:23384176

  17. The effect of high passage Mycoplasma strain F38 on the course of contagious caprine pleuropneumonia (CCPP).

    PubMed

    MacOwan, K J; Minette, J E

    1978-02-01

    In comparison with an equal number of untreated controls, goats inoculated with high passage culture of mycoplasma strain F38 were significantly less susceptible to contact infection from CCPP cases. PMID:625796

  18. Pathogenicity of Mycoplasma lipofaciens strain ML64, isolated from an egg of a Northern Goshawk (Accipiter gentilis), for chicken embryos.

    PubMed

    Lierz, M; Stark, R; Brokat, S; Hafez, H M

    2007-04-01

    Some Mycoplasma species are well-known avian pathogens and are of importance in poultry breeder flocks due to their pathogenic potential for embryos. Mycoplasmas are regularly detected in birds of prey, and a strain of Mycoplasma lipofaciens that was isolated from an egg of a Northern Goshawk (Accipiter gentilis) was examined for its pathogenicity in specific pathogen free chicken embryos since birds of prey eggs were not available for this purpose. The strain was found to be pathogenic, causing a high mortality as well as dwarfing, curled toes and infiltrations of heterophils in the liver, kidney, intestine and chorioallantoic membrane. PMID:17479376

  19. Genomic differences between type strain PG1 and field strains of Mycoplasma mycoides subsp. mycoides small-colony type

    PubMed Central

    Bischof, Daniela F.; Vilei, Edy M.; Frey, Joachim

    2006-01-01

    The recently accomplished complete genomic sequence analysis of the type strain PG1 of Mycoplasma mycoides subsp. mycoides small-colony type revealed four large repeated segments of 24, 13, 12, and 8 kb that are flanked by insertion sequence (IS) elements. Genetic analysis of type strain PG1 and African, European, and Australian field and vaccine strains revealed that the 24-kb genetic locus is repeated only in PG1 and not in other M. mycoides subsp. mycoides SC strains. In contrast, the 13-kb genetic locus was found duplicated in some strains originating from Africa and Australia but not in strains that were isolated from the European outbreaks. The 12- and 8-kb genetic loci were found in two and three copies, respectively, in all 28 strains analyzed. The flanking IS elements are assumed to lead to these tandem duplications, thus contributing to genomic plasticity. This aspect must be considered when designing novel diagnostic approaches and recombinant vaccines. PMID:16919417

  20. INITIAL PROTEOMIC ANALYSIS OF DIFFERENTIALLY EXPRESSED PROTEINS FROM MYCOPLASMA GALLISEPTICUM VACCINE STRAINS TS-11 AND F DETECTED BY WESTERN BLOTTING

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma gallisepticum (MG) reduces the number of eggs produced by layer chickens. Three live MG vaccine strains are available for use in layer chickens and include F, ts-11 and 6/85. The MG vaccine strains ts-11 and 6/85 are safer than F and they have little or no potential of spreading from bi...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  3. Immunisation of goats against contagious caprine pleuropneumonia using sonicated antigens of F-38 strain of mycoplasma.

    PubMed

    Rurangirwa, F R; Masiga, W N; Muthomi, E K

    1984-03-01

    Three groups of 15 goats each were immunised against contagious caprine pleuropneumonia (CCPP) using sonicated antigens of the F-38 strain of mycoplasma incorporated in incomplete Freund's adjuvant (IFA), emulsified in aluminium hydroxide and phosphate buffered saline respectively. Three months after immunisation, five goats from each group were challenged by the in-contact method. The goats immunised with the antigen incorporated in IFA were all solidly immune to the challenge whereas only two of five of the goats in the other two groups were protected. When the remaining 10 animals from each group were challenged six months after immunisation, those immunised with the antigen in IFA were still solidly immune while only two goats from each of the other two groups were protected. These results show that effective immunity against CCPP caused by the F-38 strain can be induced by vaccination with sonicated F-38 antigens emulsified in IFA. PMID:6718817

  4. Initial Proteomics Analysis of Differentially Expressed Proteins from Mycoplasma gallisepticum Vaccine Strains ts-11 and F Detected by Western Blotting

    Microsoft Academic Search

    2006-01-01

    Mycoplasma gallisepticum (MG) is the causative agent of chronic respiratory disease in laye r chickens. The live MG vaccine strains that are available for use in layer chickens include F, ts-11 and 6\\/85. The MG vaccine strains ts-11 and 6\\/85 are safer than F and they have little or no potential of spreading from bird to bird. However, ts-11 and

  5. Emergence of Atypical Mycoplasma agalactiae Strains Harboring a New Prophage and Associated with an Alpine Wild Ungulate Mortality Episode

    PubMed Central

    Tardy, Florence; Baranowski, Eric; Nouvel, Laurent-Xavier; Mick, Virginie; Manso-Silvàn, Lucía; Thiaucourt, François; Thébault, Patricia; Breton, Marc; Sirand-Pugnet, Pascal; Blanchard, Alain; Garnier, Alexandre; Gibert, Philippe; Game, Yvette; Poumarat, François

    2012-01-01

    The bacterium Mycoplasma agalactiae is responsible for contagious agalactia (CA) in small domestic ruminants, a syndrome listed by the World Organization for Animal Health and responsible for severe damage to the dairy industry. Recently, we frequently isolated this pathogen from lung lesions of ibexes during a mortality episode in the French Alps. This situation was unusual in terms of host specificity and tissue tropism, raising the question of M. agalactiae emergence in wildlife. To address this issue, the ibex isolates were characterized using a combination of approaches that included antigenic profiles, molecular typing, optical mapping, and whole-genome sequencing. Genome analyses showed the presence of a new, large prophage containing 35 coding sequences (CDS) that was detected in most but not all ibex strains and has a homolog in Mycoplasma conjunctivae, a species causing keratoconjunctivitis in wild ungulates. This and the presence in all strains of large integrated conjugative elements suggested highly dynamic genomes. Nevertheless, M. agalactiae strains circulating in the ibex population were shown to be highly related, most likely originating from a single parental clone that has also spread to another wild ungulate species of the same geographical area, the chamois. These strains clearly differ from strains described in Europe so far, including those found nearby, before CA eradication a few years ago. While M. agalactiae pathogenicity in ibexes remains unclear, our data showed the emergence of atypical strains in Alpine wild ungulates, raising the question of a role for the wild fauna as a potential reservoir of pathogenic mycoplasmas. PMID:22522685

  6. Efficacy of different adjuvants to potentiate the immune response to mycoplasma strain F-38.

    PubMed

    Mulira, G L; Masiga, W N; Nandokha, E

    1988-02-01

    A study was carried out to determine the efficacy of different adjuvants in enhancing antibody response to sonicated F-38 antigens. Goats were immunised against CCPP using antigens incorporated in Freund's incomplete adjuvant (IFA), saponin, aluminium hydroxide gel and buffered saline (PBS) respectively. Antibody responses were determined. The goats were challenged four months after immunisation to assess their immune status. Two of eight goats given antigen in PBS, six of 10 goats given antigen in aluminium hydroxide, seven of eight goats given antigen in IFA and all 10 goats given antigen in saponin withstood the challenge. Saponin and IFA were similar in their immune potentiation ability and were superior to aluminium hydroxide. As IFA has been considered unsuitable for use in food animals saponin may prove valuable in vaccination of goats against CCPP caused by mycoplasma strain F-38. PMID:3354056

  7. Time-dependent recovery of Mycoplasma lipofaciens (strain ML64) from incubated infertile chicken eggs and dead in shell chicken embryos.

    PubMed

    Lierz, Michael; Hafez, Hafez M

    2008-09-01

    Mycoplasmas are pathogens of different avian species, and they are able to be vertically transmitted. Even detected, Mycoplasma prevalence in raptor eggs is very low. In contrast to poultry, raptor eggs submitted for investigations are usually incubated. To investigate the influence of incubation length on the recovery of mycoplasmas from eggs, infertile specific-pathogen-free chicken eggs and embryos were infected with Mycoplasma lipofaciens (strain ML64), which had previously been isolated from an egg of a northern goshawk (Accipiter gentilis), in two different dosages. The eggs were investigated up to 12 days after infection (infertile eggs) or embryonic death. Mycoplasmas were recovered over the entire period after embryonic death by isolation. It was possible to re-isolate M. lipofaciens (strain ML64) from infertile eggs infected with 10(6) colony-forming units (CFUs) up to 12 days, but only up to 7 days if infected with 10(2) CFUs, which may be closer to the situation after natural infection. This study demonstrates that incubation of infertile eggs does have an influence on the recovery rate of mycoplasmas. This influence must be considered if interpreting results of Mycoplasma investigations in eggs of nonpoultry species. Additionally, it is recommended to use dead in shell embryos rather than infertile eggs for Mycoplasma detection. PMID:18939632

  8. Genotypic and Phenotypic Analysis of Mycoplasma fermentans Strains Isolated from Different Host Tissues

    PubMed Central

    Campo, Laura; Larocque, Patrick; La Malfa, Tiziana; Blackburn, Warren D.; Watson, Harold L.

    1998-01-01

    A correlation was found between the expression of a specific Mycoplasma fermentans surface antigen (Pra, proteinase-resistant antigen) and the site of isolation of the organism from the infected host. Strains which expressed Pra were most frequently associated with cells of bone marrow origin, and strains which lacked expression of Pra were most commonly isolated from the respiratory tract, genital tract, and arthritic joints, i.e., epithelial cell surfaces. Pra was previously shown to be resistant to degradation by proteinases and was hypothesized to play a protective role at the organism surface and perhaps to influence which host tissue site was colonized by the organism. The methods used for this phenotyping scheme required isolation and growth of the mycoplasma in quantities sufficient for immunoblot analysis using monoclonal antibodies. We wanted to determine a more rapid and less cumbersome technique to supplement this method for determining the Pra phenotype directly in clinical specimens. Here we describe PCR studies to investigate the movement of a previously identified M. fermentans insertion sequence (IS)-like element. These data showed a correlation between a specific IS genotype and the Pra+ phenotype. Production of a 160-bp product using a single set of IS-based primers was associated with expression of Pra. The genomic IS location resulting in the 160-bp product was determined by using Southern blot analysis and was found to be a stable insertion site characteristic of genotype I strains. Additional analyses of sequences within and flanking the IS insertion sites revealed another pair of PCR primer sites which resulted in the consistent production of a 450-bp amplicon. The stability of this site was dependent on the absence of the IS-like element between the primer sites. The production of this 450-bp amplicon correlated with the Pra mutant phenotype and was characteristic of genotype II strains. The data showed that the sequence within the IS may be unstable and that reliable genotyping sequences are more easily found in the stable genomic sites which flank the IS element. PMID:9574708

  9. Comparative analysis of mucosal immunity to Mycoplasma hyopneumoniae in Jiangquhai porcine lean strain and DLY piglets.

    PubMed

    Hua, L Z; Wu, Y Z; Bai, F F; William, K K; Feng, Z X; Liu, M J; Yao, J T; Zhang, X; Shao, G Q

    2014-01-01

    The Jiangquhai porcine lean strain (JQHPL) is a new pork meat-type strain that has been developed in recent years from the parent lines Duroc, Fengjing, and Jiangquhai pigs (DurocxFengjing pigxJiangquhai pig). Enzootic pneumonia (EP) in pigs induced by Mycoplasma hyopneumoniae (M. hyopneumoniae) is a chronic respiratory disease of pigs, generating high economic losses in the swine industry. Here, we investigated the degree of resistance to M. hyopneumoniae for the Jiangquhai porcine lean strain and the Duroc x Landrace x Yorkshire (DLY) pigs, which are Western commercial pigs that have been introduced in China. A total of 209 DLY piglets and 221 JQHPL piglets from 19 Landrace x Yorkshire and 22 JQHPL M. hyopneumoniae positive gestating sows with different expected dates of confinement were selected and raised in the same M. hyopneumoniae positive farrowing barn. When the oldest suckling piglets were 37 days old, nasal swabs were collected from all the piglets (ranging from 4 to 37 days old) to detect the M. hyopneumoniae pathogen using n-PCR and M. hyopneumoniae specific SIgA using ELISA. Positive M. hyopneumoniae infection rates in both the strains increased with age; however, positive rates for JQHPL were lower compared to DLY at 14 to 35 days old. The level of the specific SIgA rose rapidly in JQHPL respiratory tracts, particularly in piglets 21 to 35 days in age compared to DLY piglets of the same age; however, the level of the specific SIgA in DLY also marginally increased. In conclusion, JQHPL pigs exhibits higher resistance to M. hyopneumoniae compared to DLY. It is possible that this characteristic is caused by the faster and stronger mucosal immunity phenotype of the JQHPL strain. PMID:25061745

  10. Molecular epidemiology of contagious bovine pleuropneumonia by multilocus sequence analysis of Mycoplasma mycoides subspecies mycoides biotype SC strains

    Microsoft Academic Search

    S Lorenzon; I Arzul; A Peyraud; P Hendrikx; F Thiaucourt

    2003-01-01

    Contagious bovine pleuropneumonia is a bacterial disease caused by Mycoplasma mycoides subsp. mycoides SC (MmmSC), and included in list A of the Office International des Epizooties. It is one of the major constraints to cattle raising in sub-Saharan and south-western Africa and also a threat to all countries currently free of the disease. MmmSC strains were considered very homogeneous until

  11. Bromothymol blue broth: improved medium for detection of Ureaplasma urealyticum (T-strain mycoplasma).

    PubMed Central

    Robertson, J A

    1978-01-01

    Bromothymol blue (B) broth for the cultivation, detection, and identification of Ureaplasma urealyticum is described. In this medium, strains Cook and 960 had shorter generation times (60 min or less) and reached higher populations (over 10(8)) than have yet been reported for this species. Furthermore, the indicator changes color before the end of logarithmic growth, and the cultures retain viability for at least 1 day thereafter, greatly simplifying the handling of the organism. When the populations in cultures of these two strains and seven new isolates were determined, growth was detected earlier and proceeded to higher final titers in B broth than in urease test color medium (U-9 broth). The inclusion of antibiotics in B broth for use in clinical laboratories (B/NL broth) made the medium selective, specific, and more sensitive for the isolation of U. urealyticum. Comparison of B/NL broth with genital mycoplasma (GM) agar and U-9 broth for the primary isolation of U. urealyticum was made with 183 urethral swabs. All 70 isolates were detected on B/NL broth, but only 66 and 63 isolates were detected on GM agar and in U-9 broth, respectively. Moreover, the cultures in B/NL broth were pure and at titers that generally showed good correlation with colony counts on GM agar. PMID:632344

  12. 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

  13. Characterization of the 16S rRNA genes from Mycoplasma sp. strain F38 and development of an identification system based on PCR.

    PubMed Central

    Bascuñana, C R; Mattsson, J G; Bölske, G; Johansson, K E

    1994-01-01

    Mycoplasma sp. (strain F38) is the causative agent of contagious caprine pleuropneumonia, which is a goat disease of great global concern. Strain F38 belongs to the so-called "Mycoplasma mycoides cluster," and the members of this cluster have many biochemical and serological properties in common, which makes it difficult to differentiate between them by conventional methods. Their phylogenetic interrelationship are thus uncertain. The 16S rRNA gene of the rrnB operon from strain F38 was cloned and sequenced. The sequence was compared with the 16S rRNA sequences of related mycoplasmas, and phylogenetic trees were constructed by parsimony analysis. A three-way ambiguity among strain F38, Mycoplasma capricolum, and Mycoplasma sp. strain PG50 was observed in the trees. This observation is in agreement with a recent proposal to reclassify strain F38 and M. capricolum. A primer set was designed for in vitro amplification by PCR of a fragment of the 16S rRNA genes from the M. mycoides cluster. The amplimers of strain F38 could be distinguished easily from the corresponding amplimers from other members of the M. mycoides cluster by restriction enzyme analysis with PstI. This observation was utilized to design an identification system for strain F38. Part of the 16S rRNA gene of the rrnA operon from strain F38 was also cloned, and several sequence differences between the two rRNA operons were discovered, revealing microheterogeneity between the two 16S rRNA genes of this organism. Images PMID:8169205

  14. Identification of some clinical strains of CDC coryneform group A-3 and A-4 bacteria as Cellulomonas species and proposal of Cellulomonas hominis sp. nov. for some group A-3 strains.

    PubMed Central

    Funke, G; Ramos, C P; Collins, M D

    1995-01-01

    CDC coryneform group A-3 and A-4 bacteria were defined by Hollis and Weaver in 1981, but their taxonomic position is still unclear. By using biochemical and chemotaxonomical methods, four clinical strains belonging to CDC coryneform groups A-3 (n = 2) and A-4 (n = 2) were studied and could be assigned to the genus Cellulomonas, resulting in the first description of Cellulomonas strains isolated from clinical specimens. CDC coryneform group A-3 and A-4 strains were compared with the type strains of the seven species constituting the genus Cellulomonas at present as well as with the closely related species Oerskovia turbata, Oerskovia xanthineolytica, and Jonesia denitrificans, but their biochemical patterns were not compatible with the patterns of any of those species. Almost the entire sequences of the 16S rRNA genes of one representative strain of both CDC taxa were determined, and comparative sequence analysis confirmed the placement of the CDC coryneform group A-3 and A-4 strains studied in the Cellulomonas-Oerskovia subbranch of the actinomycetes. Both CDC taxa exhibited > 99% base pair homology within their 16S rDNAs. On the basis of phenotypic and molecular data, we formally propose a new species, Cellulomonas hominis sp. nov., for the CDC coryneform group A-3 bacteria examined. The type strain is DSM 9581. The precise taxonomic status of the CDC coryneform group A-4 strains studied remains to be established by quantitative DNA-DNA hybridizations. PMID:7559954

  15. Specificity and Strain-Typing Capabilities of Nanorod Array-Surface Enhanced Raman Spectroscopy for Mycoplasma pneumoniae Detection

    PubMed Central

    Henderson, Kelley C.; Benitez, Alvaro J.; Ratliff, Amy E.; Crabb, Donna M.; Sheppard, Edward S.; Winchell, Jonas M.; Dluhy, Richard A.; Waites, Ken B.; Atkinson, T. Prescott; Krause, Duncan C.

    2015-01-01

    Mycoplasma pneumoniae is a cell wall-less bacterial pathogen of the human respiratory tract that accounts for > 20% of all community-acquired pneumonia (CAP). At present the most effective means for detection and strain-typing is quantitative polymerase chain reaction (qPCR), which can exhibit excellent sensitivity and specificity but requires separate tests for detection and genotyping, lacks standardization between available tests and between labs, and has limited practicality for widespread, point-of-care use. We have developed and previously described a silver nanorod array-surface enhanced Raman Spectroscopy (NA-SERS) biosensing platform capable of detecting M. pneumoniae with statistically significant specificity and sensitivity in simulated and true clinical throat swab samples, and the ability to distinguish between reference strains of the two main genotypes of M. pneumoniae. Furthermore, we have established a qualitative lower endpoint of detection for NA-SERS of < 1 genome equivalent (cell/?l) and a quantitative multivariate detection limit of 5.3 ± 1 cells/?l. Here we demonstrate using partial least squares- discriminatory analysis (PLS-DA) of sample spectra that NA-SERS correctly identified M. pneumoniae clinical isolates from globally diverse origins and distinguished these from a panel of 12 other human commensal and pathogenic mycoplasma species with 100% cross-validated statistical accuracy. Furthermore, PLS-DA correctly classified by strain type all 30 clinical isolates with 96% cross-validated accuracy for type 1 strains, 98% cross-validated accuracy for type 2 strains, and 90% cross-validated accuracy for type 2V strains. PMID:26121242

  16. Molecular evolution of Mycoplasma capricolum subsp. capripneumoniae strains, based on polymorphisms in the 16S rRNA genes.

    PubMed

    Pettersson, B; Bölske, G; Thiaucourt, F; Uhlén, M; Johansson, K E

    1998-05-01

    Mycoplasma capricolum subsp. capripneumoniae belongs to the so-called Mycoplasma mycoides cluster and is the causal agent of contagious caprine pleuropneumonia (CCPP). All members of the M. mycoides cluster have two rRNA operons. The sequences of the 16S rRNA genes of both rRNA operons from 20 strains of M. capricolum subsp. capripneumoniae of different geographical origins in Africa and Asia were determined. Nucleotide differences which were present in only one of the two operons (polymorphisms) were detected in 24 positions. The polymorphisms were not randomly distributed in the 16S rRNA genes, and some of them were found in regions of low evolutionary variability. Interestingly, 11 polymorphisms were found in all the M. capricolum subsp. capripneumoniae strains, thus defining a putative ancestor. A sequence length difference between the 16S rRNA genes in a poly(A) region and 12 additional polymorphisms were found in only one or some of the strains. A phylogenetic tree was constructed by comparative analysis of the polymorphisms, and this tree revealed two distinct lines of descent. The nucleotide substitution rate of strains within line II was up to 50% higher than within line I. A tree was also constructed from individual operonal 16S rRNA sequences, and the sequences of the two operons were found to form two distinct clades. The topologies of both clades were strikingly similar, which supports the use of 16S rRNA sequence data from homologous operons for phylogenetic studies. The strain-specific polymorphism patterns of the 16S rRNA genes of M. capricolum subsp. capripneumoniae may be used as epidemiological markers for CCPP. PMID:9573185

  17. The Genome Sequence of Mycoplasma hyopneumoniae Strain 232, the Agent of Swine Mycoplasmosis

    Microsoft Academic Search

    F. Chris Minion; Elliot J. Lefkowitz; Melissa L. Madsen; Barbara J. Cleary; Steven M. Swartzell; Gregory G. Mahairas

    2004-01-01

    We present the complete genome sequence of Mycoplasma hyopneumoniae, an important member of the porcine respiratory disease complex. The genome is composed of 892,758 bp and has an average GC content of 28.6 mol%. There are 692 predicted protein coding sequences, the average protein size is 388 amino acids, and the mean coding density is 91%. Functions have been assigned

  18. Real-Time PCR Assays To Address Genetic Diversity among Strains of Mycoplasma hyopneumoniae

    Microsoft Academic Search

    Erin L. Strait; Melissa L. Madsen; F. Chris Minion; Jane Christopher-Hennings; Matthew Dammen; Katherine R. Jones; Eileen L. Thacker

    2008-01-01

    Mycoplasma hyopneumoniae is an important cause of pneumonia in pigs around the world, but confirm- ing its presence in (or absence from) pigs can be difficult. Culture for diagnosis is impractical, and seroconversion is often delayed after natural infection, limiting the use of serology. Numerous PCR assays for the detection of M. hyopneumoniae have been developed, targeting several different genes.

  19. EFFECTS OF F-STRAIN MYCOPLASMA GALLISEPTICUM INOCULATION AT TWELVE WEEKS OF AGE ON EGG YOLK COMPOSITION IN COMMERCIAL EGG LAYING HENS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In two trials, the effects of F-strain Mycoplasma gallisepticum (FMG) on the content of egg yolks from commercial Single Combed White Leghorn laying hens were investigated over a production cycle. Ten hens were assigned to each of eight (Trial 1) or sixteen (Trial 2) negative pressure fiberglass bi...

  20. Influence of supplemental dietary poultry fat on the yolk characteristics of commercial layers inoculated before or at the onset of lay with F-strain Mycoplasma gallisepticum

    Microsoft Academic Search

    E. D. Peebles; M. R. Burnham; S. L. Branton; S. K. Womack

    2009-01-01

    3 Abstract: Effects of F-strain Mycoplasma gallisepticum (FMG) inoculation and 1.5% supplemental dietary Poultry Fat (PF) on the blood characteristics of commercial layers between 24 and 58 wk of age were investigated. Sham and FMG inoculations were administered at 12 (before lay) and 22 (early in lay) wk of age and dietary treatments (Basal Control Diets (BC) and basal control

  1. Cholesterol Requirement of Mycoplasmas

    PubMed Central

    Razin, Shmuel; Tully, Joseph G.

    1970-01-01

    Cholesterol requirement for growth of mycoplasmas was tested in a serum-free medium supplemented with albumin, l-arginine, palmitic acid, and various concentrations of cholesterol dissolved in Tween 80. In cases in which Tween 80 was shown to inhibit growth, the test medium was supplemented with cholesterol dissolved in ethanol. Of the 31 species examined, all but Mycoplasma laidlawii, M. granularum, and Mycoplasma species strain S-743 exhibited a growth response to cholesterol. No requirement for cholesterol could be shown with the stable L-phase variants of Streptobacillus moniliformis and Proteus species. The results provide experimental support for the view that the large majority of the established Mycoplasma species require cholesterol for growth. PMID:4911537

  2. Measurement of the cytotoxic effects of different strains of Mycoplasma equigenitalium on the equine uterine tube using a calmodulin assay.

    PubMed Central

    Bermúdez, V M; Miller, R B; Rosendal, S; Fernando, M A; Johnson, W H; O'Brien, P J

    1992-01-01

    The cytopathic effects induced by five strains of Mycoplasma equigenitalium for cells of equine uterine tube explants were tested by measuring changes in cellular and extracellular concentrations of calmodulin (CaM). Calmodulin concentrations in samples of total homogenate (TH) and total homogenate supernates (THS) of the infected equine uterine tube explants were significantly lower than respective measurements on noninfected controls. In tissue culture medium fractions (TCM) of some infected explants, CaM concentrations were significantly higher than noninfected controls (p > 0.95). The results suggest that M. equigenitalium colonization on ciliated cells of the equine uterine tube can affect the permeability of the cell membrane leading to leakage or release of CaM during cell breakdown. Measurement of CaM concentrations in samples of TH revealed significant differences in the cytotoxic effects induced by different strains of M. equigenitalium on the equine uterine tube (EUT). The data suggests that some strains of M. equigenitalium may have a role in reproductive failure in the mare. In addition comparisons of the means of the concentrations of CaM in samples of TH or THS in EUT explants from four mares in the follicular and four in the luteal phase of the estrous cycle were found to be not significantly different. PMID:1477802

  3. The prevalence of antibody of antibody to contagious caprine pleuropneumonia (Mycoplasma strain F38) in some wild herbivores and camels in Kenya.

    PubMed

    Paling, R W; Macowan, K J; Karstad, L

    1978-07-01

    Sera of 11 species of wild herbivores were tested for antibody to Mycoplasma strain F38 which causes contagious caprine pleuropneumonia (CCPP) in Kenya. Antibodies were found in buffalo (Syncerus caffer) (32%), impala (Aepyceros melampus) (10%) and camels (Camelus dromedarius) (49%) but not in bushbuck (Tragelaphus scriptus), eland (Taurotragus oryx), Grant's gazelle (Gazella granti), kongoni (Alcelaphus buselaphus cokei), oryx (Oryx beisa), Thomson's gazelle (Gazella thomsonii), waterbuck (Kobus defassa) and wildebeest (Connochaetus taurinus). PMID:691121

  4. Spreading factors of Mycoplasma alligatoris, a flesh-eating mycoplasma.

    PubMed

    Brown, D R; Zacher, L A; Farmerie, W G

    2004-06-01

    Mycoplasma alligatoris causes lethal invasive disease of alligators and caimans. A homolog of the nagH gene, encoding a hyaluronidase secreted by Clostridium perfringens, and a C. perfringens hyaluronidase nagI or nagK pseudogene were discovered in the M. alligatoris genome. The nagH gene was detected by PCR in the closest relative of M. alligatoris, Mycoplasma crocodyli, but not in 40 other species representing the Mycoplasma hominis, Mycoplasma pneumoniae, and Spiroplasma phylogenetic clusters. The hyaluronidase activity in the cellular fraction of M. alligatoris and M. crocodyli SP4 broth cultures was equivalent to 10(-16) U of Streptomyces hyalurolyticus hyaluronidase CFU(-1). Negligible activity was present in the cell-free supernatant fraction. No chondroitinase activity was detected. There is also a novel homolog of the nanI gene, which encodes a sialidase secreted by C. perfringens, in the M. alligatoris genome. The signature YRIP and SXDXGXTW motifs and catalytic residues of the clostridial sialidase are conserved in the mycoplasmal gene, but the leader sequence necessary for its secretion by C. perfringens is absent. The gene was not detected by PCR in any other mycoplasma. Potent cell-associated sialidase activity was present in M. alligatoris colonies on agar but not in the cell-free supernatants of broth cultures or in M. crocodyli. The presence of hyaluronidase and sialidase in M. alligatoris is consistent with the rapid invasiveness and necrotizing effects of this organism, and the lack of sialidase in M. crocodyli is consistent with its comparatively attenuated virulence. This genetic and biochemical evidence suggests that the spreading factors hyaluronidase and sialidase, a combination unprecedented in mycoplasmas, are the basis of the virulence of M. alligatoris. PMID:15175306

  5. Characterization of Strains of Mycoplasma mycoides subsp. mycoides Small Colony Type Isolated from Recent Outbreaks of Contagious Bovine Pleuropneumonia in Botswana and Tanzania: Evidence for a New Biotype

    PubMed Central

    March, John B.; Clark, Jason; Brodlie, Malcolm

    2000-01-01

    Four strains of Mycoplasma mycoides subsp. mycoides small colony type (MmmSC) isolated from recent outbreaks of contagious bovine pleuropneumonia (CBPP) in Africa have been investigated. One Botswanan strain, M375, displayed numerous and significant phenotypic differences from both contemporary field isolates and older field and vaccine strains (African, Australian, and European strains dating back to 1936). Differences include altered morphology, reduced capsular polysaccharide production, high sensitivity to MmmSC rabbit hyperimmune antisera in vitro, and unique polymorphisms following immunoblotting. While insertion sequence analysis using IS1634 clearly indicates a close evolutionary relationship to west African strains, hybridization with IS1296 shows the absence of a band present in all other strains of MmmSC examined. The data suggest that a deletion has occurred in strain M375, which may explain its altered phenotype, including poor growth in vitro and a relative inability to cause septicemia in mice. These characteristics are also exhibited by Mycoplasma capricolum subsp. capripneumoniae (causal agent of contagious caprine pleuropneumonia [CCPP]), against which M375 antiserum exhibited some activity in vitro (unique among the various MmmSC antisera tested). These findings may have evolutionary implications, since CCPP is believed to be lung specific and without a septicemic phase (unlike CBPP). Since M375 was isolated from a clinical case of CBPP, this novel biotype may be fairly widespread but not normally isolated due to difficulty of culture and/or a potentially altered disease syndrome. Bovine convalescent antisera (obtained from contemporary naturally infected cattle in Botswana) were active against strain M375 in an in vitro growth inhibition test but not against any other strains of MmmSC tested. There exists the possibility therefore, that strain M375 may possess a set of protective antigens different from those of other strains of MmmSC (including vaccine strains). These findings have implications for the control of the current CBPP epidemic in Africa. PMID:10747118

  6. Characterization of strains of Mycoplasma mycoides subsp. mycoides small colony type isolated from recent outbreaks of contagious bovine pleuropneumonia in Botswana and Tanzania: evidence for a new biotype.

    PubMed

    March, J B; Clark, J; Brodlie, M

    2000-04-01

    Four strains of Mycoplasma mycoides subsp. mycoides small colony type (MmmSC) isolated from recent outbreaks of contagious bovine pleuropneumonia (CBPP) in Africa have been investigated. One Botswanan strain, M375, displayed numerous and significant phenotypic differences from both contemporary field isolates and older field and vaccine strains (African, Australian, and European strains dating back to 1936). Differences include altered morphology, reduced capsular polysaccharide production, high sensitivity to MmmSC rabbit hyperimmune antisera in vitro, and unique polymorphisms following immunoblotting. While insertion sequence analysis using IS1634 clearly indicates a close evolutionary relationship to west African strains, hybridization with IS1296 shows the absence of a band present in all other strains of MmmSC examined. The data suggest that a deletion has occurred in strain M375, which may explain its altered phenotype, including poor growth in vitro and a relative inability to cause septicemia in mice. These characteristics are also exhibited by Mycoplasma capricolum subsp. capripneumoniae (causal agent of contagious caprine pleuropneumonia [CCPP]), against which M375 antiserum exhibited some activity in vitro (unique among the various MmmSC antisera tested). These findings may have evolutionary implications, since CCPP is believed to be lung specific and without a septicemic phase (unlike CBPP). Since M375 was isolated from a clinical case of CBPP, this novel biotype may be fairly widespread but not normally isolated due to difficulty of culture and/or a potentially altered disease syndrome. Bovine convalescent antisera (obtained from contemporary naturally infected cattle in Botswana) were active against strain M375 in an in vitro growth inhibition test but not against any other strains of MmmSC tested. There exists the possibility therefore, that strain M375 may possess a set of protective antigens different from those of other strains of MmmSC (including vaccine strains). These findings have implications for the control of the current CBPP epidemic in Africa. PMID:10747118

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

    PubMed

    Branton, S L; Leigh, S A; Purswell, J L; Evans, J D; Collier, S D; Olanrewaju, H A; Pharr, G T

    2010-09-01

    Vaccination of multi-age layer operations, wherein one million plus commercial layer chickens are housed, has been spurious until the development of a self-propelled, constant-speed spray vaccinator. Still, even with its use, live Mycoplasma gallisepticum (MG) vaccinations have been questionable in terms of seroconversion. Using the vaccinator as a research tool over the past 5 yr, factors have been elucidated which impact seroconversion to one live MG vaccine in particular, the F strain of MG (FMG). These factors include the type of nozzle used to spray the vaccine, the temperature of the water used to rehydrate and administer the vaccine, and the pH and osmolarity of the fluid used to apply the vaccine. In the present study, one farm was monitored for its seroconversion rates over 4 1/2 yr, during which time the FMG vaccination protocol was amended as factors were identified that enhanced seroconversion rates. The results of this study showed that implementation and inclusion of the optimized factors into the vaccination protocol for FMG enhanced seroconversion rates because they went from an initial 50%-55% positive seroconversion rate to a consistent 100% positive seroconversion rate over the 56-mo study period. PMID:20945798

  8. Genetic diversity and evolution of Mycoplasma capricolum subsp. capripneumoniae strains from eastern Africa assessed by 16S rDNA sequence analysis.

    PubMed

    Heldtander, M; Wesonga, H; Bölske, G; Pettersson, B; Johansson, K E

    2001-01-01

    Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae), the causal agent of contagious caprine pleuropneumonia (CCPP), is a member of the so-called Mycoplasma mycoides cluster. These mycoplasmas have two rRNA operons in which intraspecific variations have been demonstrated. The sequences of the 16S rRNA genes of both operons from 13 field strains of M. capripneumoniae from three neighbouring African countries (Kenya, Ethiopia, and Tanzania) were determined. Four new and unique polymorphism patterns reflecting the intraspecific variations were found. Two of these patterns included length differences between the rrnA and rrnB operons. The length difference in one of the patterns was caused by a two-nucleotide insert (TG) in the rrnB operon and the length difference in the other pattern was due to a three-nucleotide deletion, also in the rrnB operon. Another pattern was characterised by a polymorphic position caused by a mutation that is known to cause streptomycin resistance in other bacterial species. The strain with this pattern was also found to be resistant to streptomycin. Streptomycin resistant clones were selected from four M. capripneumoniae strains to further investigate the correlation of this mutation to streptomycin resistance. Mutations in the 16S rRNA genes had occurred in two of these strains. The fourth pattern included a new polymorphism in position 1059. The results show that polymorphisms in M. capripneumoniae strains can be used as epidemiological markers for CCPP in smaller geographical areas and to study the molecular evolution of this species. PMID:11118738

  9. Genetic evolution of Mycoplasma capricolum subsp. capripneumoniae strains and molecular epidemiology of contagious caprine pleuropneumonia by sequencing of locus H2.

    PubMed

    Lorenzon, S; Wesonga, H; Ygesu, Laikemariam; Tekleghiorgis, Tesfaalem; Maikano, Y; Angaya, M; Hendrikx, P; Thiaucourt, F

    2002-03-01

    Contagious caprine pleuropneumonia (CCPP) is a major threat to goat farming in developing countries. Its exact distribution is not well known, despite the fact that new diagnostic tools such as PCR and competitive ELISA are now available. The authors developed a study of the molecular epidemiology of the disease, based on the amplification of a 2400 bp long fragment containing two duplicated gene coding for a putative membrane protein. The sequence of this fragment, obtained on 19 Mycoplasma capricolum subsp. capripneumoniae (Mccp) strains from various geographical locations, gave 11 polymorphic positions. The three mutations found on gene H2prim were silent and did not appear to induce any amino acid modifications in the putative translated protein. The second gene may be a pseudogene not translated in vivo, as it bore a deletion of the ATG codon found in the other members of the "Mycoplasma mycoides cluster" and as the six mutations evidenced in the Mccp strains would induce modifications in the translated amino acids. In addition, an Mccp strain isolated in the United Arab Emirates showed a deletion of the whole pseudogene, a further indication that this gene is not compulsory for mycoplasma growth. Four lineages were defined, based on the nucleotide sequence. These correlated relatively well with the geographical origin of the strains: North, Central or East Africa. The strain of Turkish origin had a sequence similar to that found in North African strains, while strains isolated in Oman had sequences similar to those of North or East African strains. The latter is possibly due to the regular import of goats of various origins. Similar molecular epidemiology tools have been developed by sequencing the two operons of the 16S rRNA gene or by AFLP. All these various techniques give complementary results. One (16S rRNA) offers the likelihood of a finer identification of strains circulating in a region, another (H2) of determining the geographical origin of the strains. These tools can make a very useful contribution to understanding the epidemiology of CCPP. PMID:11844618

  10. Comparison of immune responses to intranasal and intrapulmonary vaccinations with the attenuated Mycoplasma hyopneumoniae 168 strain in pigs.

    PubMed

    Li, Pengcheng; Li, Yunfeng; Shao, Guoqing; Yu, Qinghua; Yang, Qian

    2015-06-01

    The aim of this study was to evaluate the immune responses to intranasal and intrapulmonary vaccinations with the attenuated Mycoplasma hyopneumoniae (Mhp) 168 strain in the local respiratory tract in pigs. Twenty-four pigs were randomly divided into 4 groups: an intranasal immunization group, an intrapulmonary immunization group, an intramuscular immunization group and a control group. The levels of local respiratory tract cellular and humoral immune responses were investigated. The levels of interleukin (IL)-6 in the early stage of immunization (P<0.01), local specific secretory IgA (sIgA) in nasal swab samples (P<0.01); and IgA- and IgG-secreting cells in the nasal mucosa and trachea were higher after intranasal vaccination (P<0.01) than in the control group. Interestingly, intrapulmonary immunization induced much stronger immune responses than intranasal immunization. Intrapulmonary immunization also significantly increased the secretion of IL-6 and local specific sIgA and the numbers of IgA- and IgG-secreting cells. The levels of IL-10 and interferon-? in the nasal swab samples and the numbers of CD4(+) and CD8(+) T lymphocytes in the lung and hilar lymph nodes were significantly increased by intrapulmonary immunization compared with those in the control group (P<0.01). These data suggest that intrapulmonary immunization with attenuated Mhp is effective in evoking local cellular and humoral immune responses in the respiratory tract. Intrapulmonary immunization with Mhp may be a promising route for defense against Mhp in pigs. PMID:25649413

  11. Comparison of immune responses to intranasal and intrapulmonary vaccinations with the attenuated Mycoplasma hyopneumoniae 168 strain in pigs

    PubMed Central

    LI, Pengcheng; LI, Yunfeng; SHAO, Guoqing; YU, Qinghua; YANG, Qian

    2015-01-01

    The aim of this study was to evaluate the immune responses to intranasal and intrapulmonary vaccinations with the attenuated Mycoplasma hyopneumoniae (Mhp) 168 strain in the local respiratory tract in pigs. Twenty-four pigs were randomly divided into 4 groups: an intranasal immunization group, an intrapulmonary immunization group, an intramuscular immunization group and a control group. The levels of local respiratory tract cellular and humoral immune responses were investigated. The levels of interleukin (IL)-6 in the early stage of immunization (P<0.01), local specific secretory IgA (sIgA) in nasal swab samples (P<0.01); and IgA- and IgG-secreting cells in the nasal mucosa and trachea were higher after intranasal vaccination (P<0.01) than in the control group. Interestingly, intrapulmonary immunization induced much stronger immune responses than intranasal immunization. Intrapulmonary immunization also significantly increased the secretion of IL-6 and local specific sIgA and the numbers of IgA- and IgG-secreting cells. The levels of IL-10 and interferon-? in the nasal swab samples and the numbers of CD4+ and CD8+ T lymphocytes in the lung and hilar lymph nodes were significantly increased by intrapulmonary immunization compared with those in the control group (P<0.01). These data suggest that intrapulmonary immunization with attenuated Mhp is effective in evoking local cellular and humoral immune responses in the respiratory tract. Intrapulmonary immunization with Mhp may be a promising route for defense against Mhp in pigs. PMID:25649413

  12. 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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  13. Molecular typing of Iranian field isolates Mycoplasma synoviae and their differentiation from the live commercial vaccine strain MS-H using vlhA gene.

    PubMed

    Bayatzadeh, Mohammad Ali; Pourbakhsh, Seyed Ali; Ashtari, Abass; Abtin, Ali Reza; Abdoshah, Mohammad

    2014-01-01

    1. The single-copy domain of the N-terminal region of the vlhA gene of Mycoplasma synoviae was sequenced, analysed and verified and used to type Iranian field isolates of M. synoviae and the MS-H live vaccine strain. In addition, a restriction fragment length polymorphism (RFLP) method was developed to differentiate between field isolates of Iranian and MS-H vaccine strains. 2. All sequences were analysed and aligned; the percentage similarity of the DNA was calculated and dendrograms were constructed. Based on single nucleotide polymorphism (SNP) that existed in all field isolates in Iran, the PCR-RFLP method allowed the differentiation of all M. synoviae field isolates from the vaccine strain. 3. Using phylogenetic analysis, the isolates were assigned to 8 unique genotypes and, within each group, DNA had a high level of similarity. 4. DNA sequence analysis and PCR-RFLP of the amplicon based on percent similarity and evolutionary relationship appeared to be useful tools for strain differentiation whether M. synoviae clinical isolates from infected chickens were derived from the vaccine strain or wild-type strains. 5. This study confirms the potential value of strain typing for epidemiological purposes and suggests that phylogenetic studies are essential to understand the true relationships between strains. PMID:24405029

  14. 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. PMID:25970590

  15. Influence of Supplemental Dietary Poultry Fat on the Digestive and Reproductive Organ Characteristics of Commercial Layers Inoculated Before or at the Onset of Lay with F-Strain Mycoplasma gallisepticum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects of F-strain Mycoplasma gallisepticum (FMG) inoculation and 1.5 % supplemental dietary poultry fat (PF) on the digestive and reproductive organ characteristics of commercial layers at 58 wk of age were investigated. Sham and FMG inoculations were administered at 12 (before lay) and 22 (e...

  16. High rates of genital mycoplasma infection in the highlands of Papua New Guinea determined both by culture and by a commercial detection kit.

    PubMed Central

    Clegg, A; Passey, M; Yoannes, M; Michael, A

    1997-01-01

    Duplicate vaginal swabs were collected from 100 women, and comparisons were made between an in-house broth-agar culture system and a commercially available kit, the Mycoplasma IST kit (bioMérieux), for the detection of Mycoplasma hominis and Ureaplasma urealyticum. There was good agreement between the two systems for detection of the genital mycoplasmas in terms of sensitivity, with values of > 92% being obtained. In terms of specificity, the mutual comparisons were less favorable, though specificity values of > 72% were obtained. Statistically there was no significant difference in the performance of the two tests (P < 0.1 for both M. hominis and U. urealyticum). While the broth-agar culture system was considerably less expensive than the kit, the Mycoplasma IST kit provided additional information on antibiotic susceptibilities and had the advantages of a shelf life of up to 12 months and not requiring the preparation of culture media. The prevalences of colonization obtained for M. hominis and U. urealyticum were extremely high in this randomly selected group of women from periurban and rural settlements in the Eastern Highlands of Papua New Guinea, being > or = 70% for M. hominis and > or = 78% for U. urealyticum. colonization with both genital mycoplasmas simultaneously was also very common, with > or = 60% of women being colonized by both M. hominis and U. urealyticum. PMID:8968907

  17. 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

  18. Clinical significance of Blastocystis hominis.

    PubMed Central

    Qadri, S M; al-Okaili, G A; al-Dayel, F

    1989-01-01

    A total of 19,252 stool specimens from 12,136 patients were examined by direct microscopy and the ethyl acetate-Formalin concentration method during the last 2 years. All liquid specimens and those in which parasite identification was difficult or equivocal were also examined in trichrome-stained preparations. A total of 3,070 intestinal parasites were seen in 2,889 patients. Blastocystis hominis was found in fecal material from 647 patients (17.5%). A total of 132 cases (25.6%) were observed to be in association with other enteric pathogens. B. hominis in large numbers was present as the only parasite or with other commensals in 515 specimens from patients (79.6%). Of these patients, 239 (46.4%) had symptoms, the most common being abdominal pain (87.9%), constipation (32.2%), diarrhea (23.4%), alternating diarrhea and constipation (14.5%), vomiting (12.5%), and fatigue (10.5%). Forty-three (18%) of the patients were treated with metronidazole (0.5 to 1.0 g/day) because of recurrent symptoms and the presence of large numbers of B. hominis cells in repeated stool specimens. After 7 to 10 days of treatment, all patients became asymptomatic with negative stools on follow-up examinations for B. hominis. PMID:2808664

  19. Characterization of Mycoplasmas isolated from pneumonic lungs of sheep and goats

    Microsoft Academic Search

    R. K. Adehan; A. T. P. Ajuwape; A. I. Adetosoye; O. O. Alaka

    2006-01-01

    Fifty Mycoplasma strains were recovered from pneumonic lungs of sheep and goats from Cotonou abattoirs. Biochemically, Mycoplasma strains from sheep were divided into four groups namely: (A) eight strains which hydrolysed glucose and reduced tetrazolium chloride; (B) four strains which hydrolysed glucose, digested serum, reduced tetrazolium chloride and showed phosphatase activity; (C) four strains which hydrolysed glucose, reduced tetrazolium chloride

  20. Urogenital Mycoplasmas and Human Papilloma Virus in Hemodialysed Women

    PubMed Central

    Ekiel, Alicja; Pietrzak, Bronis?awa; Aptekorz, Ma?gorzata; Mazanowska, Natalia; Kami?ski, Pawe?; Martirosian, Gayane

    2013-01-01

    Bacterial infections, especially endogenous, are the frequent complications among hemodialyzed and renal transplant patients. In this study we assumed the prevalence of urogenital mycoplasmas and HPV among hemodialysed women. We examined 32 hemodialysed women aged 20–48 (mean 35.6 ± 8.23) and 100 healthy controls of the same ages. Two swabs were collected for detection of mycoplasmas and HPV. Culture of Ureaplasma spp. and M. hominis was performed using Mycoplasma IST2 (bioMérieux, France), Identificaton of U. parvum and U. urealyticum was performed by Kong. Primers described by Jensen were used for M. genitalium. For detection of high-risk HPV types Amplicor HPV (Roche Molecular System, CA) was used. Prevalence of urogenital mycoplasmas in the hemodialysed women (53.1%) was significantly higher (P = 0.0059), compared with controls (25%). In both groups, U. parvum was the most frequently isolated. Cooccurrence of urogenital mycoplasmas was shown in 75% of the HPV-positive hemodialysed women and in 30.4% of HPV-positive controls (P = 0.0461). Cooccurrence of urogenital mycoplasmas with HPV was significantly higher in hemodialysed women. The need to take into account these microorganisms in routine diagnostic, especially for hemodialysed patients, was demonstrated. Further studies to demonstrate the role of this cooccurrence in etiopathogenesis of infection in hemodialysed patients are required. PMID:24363622

  1. Mycoplasma agassizii Strain Variation and Distinct Host Antibody Responses Explain Differences between Enzyme-Linked Immunosorbent Assays and Western Blot Assays ?

    PubMed Central

    Wendland, Lori D.; Klein, Paul A.; Jacobson, Elliott R.; Brown, Mary B.

    2010-01-01

    The precarious status of desert (Gopherus agassizii) and gopher (G. polyphemus) tortoises has resulted in conservation efforts that now include health assessment as an important component of management decision-making. Mycoplasmal upper respiratory tract disease (URTD) is one of very few diseases in chelonians for which comprehensive and rigorously validated diagnostic tests exist. In this study, serum samples obtained from eight Gopherus tortoises documented at necropsy to (i) be enzyme-linked immunosorbent assay (ELISA) seropositive using the PS6 antigen, (ii) be infected with Mycoplasma agassizii as indicated by direct isolation of the pathogen from the respiratory surfaces, and (iii) have histological lesions of mycoplasmal URTD were used to evaluate four distinct clinical isolates of M. agassizii as antigens for ELISA and Western blot analyses. Each animal sample reacted in the Western blot with its homologous M. agassizii strain, but recognition of heterologous M. agassizii strains was variable. Further, individual animals varied significantly with respect to the specific proteins recognized by the humoral immune response. An additional 114 Gopherus serum samples were evaluated using ELISA antigens prepared from the four distinct M. agassizii strains; A405 values were significantly correlated (r2 goodness of fit range, 0.708 to 0.771; P < 0.0001) for all antigens tested. The results confirm that strain variation is responsible for the observed differences between Western blot binding patterns. Thus, reliance on a single M. agassizii strain as an antigen in Western blot assays may provide false-negative results. This could have adverse consequences for the well-being of these environmentally sensitive hosts if false-negative animals were relocated to sites consisting of true-negative populations. PMID:20810678

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

    Microsoft Academic Search

    Irina Gerchman; Inna Lysnyansky; Shimon Perk; Sharon Levisohn

    2008-01-01

    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,

  3. Effects of intrauterine infection by Staphylococcus aureus and Mycoplasma capricolum on the fertility of Nubian goats

    E-print Network

    Paris-Sud XI, Université de

    Effects of intrauterine infection by Staphylococcus aureus and Mycoplasma capricolum injection of either a Staphylococcus aureus or a Mycoplasma capricolum strain of caprine origin. A third of Staphylococcus aureus and Mycoplasma capricolum on corpus luteum regression and its consequences on reproductive

  4. Susceptibilities of genital mycoplasmas to the newer quinolones as determined by the agar dilution method.

    PubMed

    Kenny, G E; Hooton, T M; Roberts, M C; Cartwright, F D; Hoyt, J

    1989-01-01

    The increasing resistance of genital mycoplasmas to tetracycline poses a problem because tetracycline is one of the few antimicrobial agents active against Mycoplasma hominis, Ureaplasma urealyticum, chlamydiae, gonococci, and other agents of genitourinary-tract disease. Since the quinolones are a promising group of antimicrobial agents, the susceptibilities of M. hominis and U. urealyticum to the newer 6-fluoroquinolones were determined by the agar dilution method. Ciprofloxacin, difloxacin, and ofloxacin had good activity against M. hominis, with the MIC for 50% of isolates tested (MIC50) being 1 microgram/ml. Fleroxacin, lomefloxacin, pefloxacin, and rosoxacin had MIC50s of 2 micrograms/ml. Enoxacin, norfloxacin, and amifloxacin had MIC50s of 8 to 16 micrograms/ml, and cinoxacin and nalidixic acid were inactive (MIC50, greater than or equal to 256 micrograms/ml). Overall, the activities of 6-fluoroquinolones for ureaplasmas were similar to those for M. hominis, with MICs being the same or twofold greater. The most active 6-fluoroquinolones against ureaplasmas were difloxacin, ofloxacin, and pefloxacin, with MIC50s of 1 to 2 micrograms/ml. Ciprofloxacin was unusual in that the MIC50 for M. hominis was 1 microgram/ml, whereas the MIC50 for ureaplasmas was 8 micrograms/ml. Since the MIC50s for the most active quinolones approximate achievable concentrations in blood and urine, quinolones have promise in treating mycoplasmal infections. PMID:2712541

  5. Development and clinical application of an InvaderPlus(®) assay for the detection of genital mycoplasmas.

    PubMed

    Takanashi, Masaki; Ito, Shin; Kaneto, Hiroyuki; Tanahashi, Yoshikatsu; Kitanohara, Masataka; Yanagihara, Akira; Nakazima, Haruhiko; Yasuda, Mitsuru

    2015-07-01

    We developed a PCR-based assay involving Invader(®) technology for detection of the genital mycoplasmas of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum. We compared its performance with that of a PCR-microtiter plate hybridization assay, which we developed previously, in detecting genital mycoplasmas in first-voided urine (FVU) specimens from men with non-gonococcal urethritis. The tests targeting each of the genital mycoplasmas were specific for the respective species and could detect as few as 10 copies of the plasmids containing the target genes of each of the genital mycoplasmas per reaction. The assay using the InvaderPlus(®) method (InvaderPlus(®) assay) showed very similar performance to that of the PCR-microtiter plate hybridization assay for detecting the genital mycoplasmas in the FVU specimens. In addition, the PCR and endonuclease reaction in the InvaderPlus(®) assay were carried out simultaneously in one procedure, thus simplifying the assay, leading to time- and labor-savings and a decrease in the risk of specimen contamination. The InvaderPlus(®) assay could be useful in diagnosing genitourinary tract infections caused by the genital mycoplasmas. PMID:25892209

  6. Vaccination of BALB/c Mice with an Avirulent Mycoplasma pneumoniae P30 Mutant Results in Disease Exacerbation upon Challenge with a Virulent Strain

    PubMed Central

    Szczepanek, S. M.; Majumder, S.; Sheppard, E. S.; Liao, X.; Rood, D.; Tulman, E. R.; Wyand, S.; Krause, D. C.; Silbart, L. K.

    2012-01-01

    Mycoplasma pneumoniae is a significant human respiratory pathogen that causes high morbidity worldwide. No vaccine to prevent M. pneumoniae infection currently exists, since the mechanisms of pathogenesis are poorly understood. To this end, we constructed a P30 cytadhesin mutant (P-130) with a drastically reduced capacity for binding to erythrocytes and an inability to glide on glass substrates. This mutant was determined to be avirulent and cannot survive in the lungs of BALB/c mice. We also ascertained that the previously identified P30 gliding motility mutant II-3R is avirulent and also cannot be recovered from the lungs of mice after infection. Mutant P130 was then assessed for its efficacy as a live attenuated vaccine candidate in mice after challenge with wild-type M. pneumoniae. After vaccination with the P-130 P30 mutant, mice showed evidence of exacerbated disease upon subsequent challenge with the wild-type strain PI1428, which appears to be driven by a Th17 response and corresponding eosinophilia. Our results are in accordance with other reports of vaccine-induced disease exacerbation in rodents and emphasize the need to better understand the basic mechanisms of M. pneumoniae pathogenesis. PMID:22252865

  7. Vaccination of BALB/c mice with an avirulent Mycoplasma pneumoniae P30 mutant results in disease exacerbation upon challenge with a virulent strain.

    PubMed

    Szczepanek, S M; Majumder, S; Sheppard, E S; Liao, X; Rood, D; Tulman, E R; Wyand, S; Krause, D C; Silbart, L K; Geary, S J

    2012-03-01

    Mycoplasma pneumoniae is a significant human respiratory pathogen that causes high morbidity worldwide. No vaccine to prevent M. pneumoniae infection currently exists, since the mechanisms of pathogenesis are poorly understood. To this end, we constructed a P30 cytadhesin mutant (P-130) with a drastically reduced capacity for binding to erythrocytes and an inability to glide on glass substrates. This mutant was determined to be avirulent and cannot survive in the lungs of BALB/c mice. We also ascertained that the previously identified P30 gliding motility mutant II-3R is avirulent and also cannot be recovered from the lungs of mice after infection. Mutant P130 was then assessed for its efficacy as a live attenuated vaccine candidate in mice after challenge with wild-type M. pneumoniae. After vaccination with the P-130 P30 mutant, mice showed evidence of exacerbated disease upon subsequent challenge with the wild-type strain PI1428, which appears to be driven by a Th17 response and corresponding eosinophilia. Our results are in accordance with other reports of vaccine-induced disease exacerbation in rodents and emphasize the need to better understand the basic mechanisms of M. pneumoniae pathogenesis. PMID:22252865

  8. Effects of 6/85-strain Mycoplasma gallisepticum inoculation alone at 10 weeks of age or in conjunction with F-strain Mycoplasma gallisepticum inoculation overlays at 22 or 45 weeks of age on the performance of commercial ....

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects of 6/85-strain M. gallisepticum (6/85MG) inoculation alone or in conjunction with a F-strain M. gallisepticum (FMG) over-lay and its timing on the performance of commercial egg laying hens were investigated. Control birds received sham inoculations at 10 wk of age. A second treated gro...

  9. Evaluation of Mycoplasma inactivation during production of biologics: egg-based viral vaccines as a model.

    PubMed

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

    2010-05-01

    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 >or=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 viral purification processes used for the manufacture of an inactivated egg-based vaccine. PMID:20228111

  10. Prevalence of genital mycoplasmas in asymptomatic male partners of women diagnosed as having chlamydial infections.

    PubMed

    Ito, Shin; Kikuchi, Mina; Seike, Kensaku; Tsuchiya, Tomohiro; Yasuda, Mitsuru; Yokoi, Shigeaki; Nakano, Masahiro; Deguchi, Takashi

    2014-02-01

    We examined 209 asymptomatic male partners of women diagnosed as having chlamydial infections for the prevalence of Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, and Ureaplasma parvum in their first-voided urine (FVU) by nucleic acid amplification tests. Quantification of leukocytes in FVU was performed by automated urine particle analyzers. Two (1.0%) men were positive for N. gonorrhoeae, and 92 (44.0%) were positive for C. trachomatis. In men negative for these pathogens, prevalences of M. genitalium, M. hominis, U. urealyticum, and U. parvum were 0.9%, 29.6%, 27.8%, and 20.1%, respectively, and 58.3% were positive for at least one species of the genital mycoplasmas. Leukocyte counts in FVU from 92 men positive for C. trachomatis were significantly greater than those from 115 men negative for C. trachomatis (p < 0.0001). However, there was no significant difference in leukocyte counts between 66 men positive for at least one species of M. hominis, U. urealyticum, and U. parvum and 48 men negative for all the species (p = 0.1657). The present population of asymptomatic male partners of women diagnosed as having chlamydial infections showed a low prevalence of M. genitalium infections but would be at high risk of being infected by the other genital mycoplasmas. However, it was still unclear whether these genital mycoplasmas would contribute to the development of inflammation of the male urethra. When these partners are negative for C. trachomatis and N. gonorrhoeae, the recommendation to presumptively treat them to disrupt transmission networks of the genital mycoplasmas would seem premature. PMID:24486047

  11. 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 ?, ?, ? and ? 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

  12. Epidemiology and pathogenicity of Blastocystis hominis.

    PubMed

    Doyle, P W; Helgason, M M; Mathias, R G; Proctor, E M

    1990-01-01

    A prospective study was performed on a large outpatient population to evaluate the epidemiology and pathogenicity of Blastocystis hominis. Patients with stool specimens positive for B. hominis and negative for other bacterial and parasitic pathogens were sent a questionnaire and were requested to submit a follow-up specimen for ova-and-parasite examination. B. hominis was identified in 530 of 16,545 specimens (3.2%). There was a spectrum of clinical-pathological presentations in the 143 patients evaluated. An asymptomatic carrier state was seen in 19 patients. Fifteen patients had an illness consistent with acute self-limited B. hominis gastroenteritis, and 21 patients had chronic gastroenteritis associated with B. hominis. In the epidemiological evaluation of 130 patients, the most common symptoms were watery diarrhea, abdominal pain, and gas. We did not find a statistically significant association between the number of organisms present and the disease state. In summary, our results are consistent with a role for B. hominis in acute and chronic gastroenteritis; however, further detailed studies are necessary to determine whether that role is one of association or causation. PMID:2298869

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  14. Eradication of Mycoplasma contaminations from cell cultures.

    PubMed

    Uphoff, Cord C; Drexler, Hans G

    2014-01-01

    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. Curr. Protoc. Mol. Biol. 106:28.5.1-28.5.12. © 2014 by John Wiley & Sons, Inc. PMID:24733241

  15. Quantitative Assessment of Mycoplasma Hemadsorption Activity by Flow Cytometry

    PubMed Central

    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 Kd. 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. PMID:24498118

  16. 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 ?MGA0674 mutant (P1H9) in vivo and observed reduced recovery and attenuated virulence in the tracheas of experimentally infected chickens. The virulence of two additional Rlow ?MGA0674 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

  17. Blastocystis hominis--past and future.

    PubMed Central

    Zierdt, C H

    1991-01-01

    The history of B. hominis is unique. Few infectious agents have provoked the many misconceptions that plague this enigmatic parasitic ameba. Conflicting descriptions of its nature and pathogenesis have continued throughout the 20th century. As seen by the greatly expanded number of reports in recent years, B. hominis is now a major subject of study, particularly for evidence of disease causation. Physicians are treating patients with intestinal disease caused by B. hominis. Many mild cases resolve in about 3 days without treatment, but others are acute and chronic disease is common. As with E. histolytica, the carrier state is often seen without symptoms. Treatment is usually with metronidazole, but emetine (for refractory infections), trimethoprim-sulfamethoxazole, and pentamidine are also effective. In fecal samples, this complex protozoan appears in a variety of cell forms which makes microscopic diagnosis difficult. As yet, no specific fluorescent-antibody test is available for diagnosis. A culture method to demonstrate the more easily recognized CB form is available, but probably not feasible for most diagnostic laboratories. The common cell forms are the CB form, the granular (mitochondria) form, and the ameba form. The unexpected size range of these forms in clinical material, from yeast size (ca. 7 microns) to giant cells of 20 to 40 microns, makes diagnosis difficult Pseudopodia may be demonstrated by the ameba form in heated microscope stage culture chambers. The anaerobic B. hominis has no cyst form. Its mitochondria are uniquely anaerobic and have no cytochrome protein or oxidative mitochondrial enzymes. Because of its many cell forms and anaerobic mitochondria, B. hominis is an organism of great interest for morphologic and biochemical study. Reproduction is asexual, usually by binary fission. Shizogony occurs in cultured cells. The CB appears to be an organelle whose specific purpose is for reproduction by shizogony. From 2 to 30 progeny are derived from schizogony. The ameba form reproduces by plasmotomy; it has no CB. The pathology of B. hominis infections has been studied in gnotobiotic guinea pigs in which inflammation of the intestinal mucosa and invasion of the superficial layers were seen. Only limited studies of human pathology are available. Those who have studied mucosal histopathology report inflammation and cellular changes that resolve after treatment. More study in this area is strongly indicated (32, 44, 57, 62, 67, 75). Ultrastructural details of B. hominis major forms, except for the schizont, are complete. The organism has no cell wall. The concentric CB takes up as much as 95% of the cell. The major organelles, which include multiple nuclei, Golgi apparatus, mitochondria, endoplasmic reticulum, fat, and other inclusions, are confined in two or four opposed pods in a thin band of peripheral cytoplasm between the spherical entire plasma membrane and the CB membrane. The pods buldge the CB membrane inward. There is evidence of a bacteroid endosymbiont. Education about B. hominis is needed. Entry of recent findings into new textbooks is imperative for its understanding among medical practitioners. Laboratory workers need to be aware of it for many reasons. The College of American Pathologists includes B. hominis in its proficiency testing samples and requires that it be reported from clinical samples. Images PMID:2004348

  18. Draft Genome Sequences of Mycoplasma alkalescens, Mycoplasma arginini, and Mycoplasma bovigenitalium, Three Species with Equivocal Pathogenic Status for Cattle

    PubMed Central

    Tardy, Florence; Baranowski, Eric; Barré, Aurélien; Blanchard, Alain; Breton, Marc; Couture, Carole; Citti, Christine; Dordet-Frisoni, Emilie; Dupuy, Virginie; Gaurivaud, Patrice; Jacob, Daniel; Lemaitre, Claire; Nikolski, Macha; Nouvel, Laurent-Xavier; Poumarat, François; Thébault, Patricia; Theil, Sébastien; Thiaucourt, François; Sirand-Pugnet, Pascal

    2013-01-01

    We report here the draft genome sequences of Mycoplasma alkalescens, Mycoplasma arginini, and Mycoplasma bovigenitalium. These three species are regularly isolated from bovine clinical specimens, although their role in disease is unclear. PMID:23766408

  19. Molecular diversity of Scottish Cryptosporidium hominis isolates.

    PubMed

    Deshpande, A; Alexander, C L; Coyne, M; Brownlie, S; Smith-Palmer, A; Jones, B L

    2015-04-01

    Cryptosporidium hominis is one of the most prevalent protozoan parasites to infect humans where transmission is via the consumption of infective oocysts. This study describes sporadic cases in addition to the molecular diversity of outbreak cases in Scotland using the glycoprotein-60 subtyping tool. From a total of 187 C. hominis isolates, 65 were subjected to further molecular analysis and 46 were found to be the common IbA10G2 subtype. Unusual subtypes included four isolates belonging to the Ia family (IaA14R3, n = 12; IaA14R2, n = 1; IaA9G3, n = 1; IaA25R3, n = 2), two from the Id family (IdA24, n = 1; IdA17, n = 1) and one belonging to the Ie family, namely IeA11G3T3. These data contribute significantly to our knowledge and understanding of the molecular diversity of C. hominis isolates from outbreak investigations involving Scottish residents which will be beneficial for the management of future outbreaks. PMID:25185671

  20. Functional Characterization of the RuvB Homologs from Mycoplasma pneumoniae and Mycoplasma genitalium?

    PubMed Central

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

    2011-01-01

    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 [RuvBFH] and M. genitalium [RuvBMge], respectively) are described. Both RuvBFH and RuvBMge 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 RuvBMge, however, was significantly lower than that of RuvBFH. Interestingly, we found RuvBFH 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 [RuvBM129]) that differs from RuvBFH in a single amino acid residue (at position 140). In contrast to RuvBFH, RuvBM129 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. PMID:21949077

  1. Mycoplasma bovis research update

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  2. Proteolytic Processing of the Mycoplasma hyopneumoniae Cilium Adhesin

    Microsoft Academic Search

    Steven P. Djordjevic; Stuart J. Cordwell; Michael A. Djordjevic; Jody Wilton; F. Chris Minion

    2004-01-01

    Mycoplasma hyopneumoniae is an economically significant swine pathogen that colonizes the respiratory ciliated epithelial cells. Cilium adherence is mediated by P97, a surface protein containing a repeating element (R1) that is responsible for binding. Here, we show that the cilium adhesin is proteolytically processed on the surface. Proteomic analysis of strain J proteins identified cleavage products of 22, 28, 66,

  3. Mycoplasmas and their role as rodent pathogens

    Microsoft Academic Search

    R. J. Fallon

    1967-01-01

    SUMMARY Mycoplasmas are the smallest known free-living form of life, and differ from bacteria in a number of characteristics. They are widely distributed in the animal kingdom and may give rise to both acute and latent infections as well as being present as normal flora. The three principal rodent pathogens so far described are Mycoplasma pulmonis, Mycoplasma arthritidis and Mycoplasma

  4. pH-controlled continuous cultivation of mycoplasmas.

    PubMed Central

    Krebs, B; Schütz, M; Fischer, M; Sommer, G; Kirchhoff, H

    1989-01-01

    The continuous cultivation of mycoplasmas in a pH-controlled metabolistat was investigated with the fermentative strain Mycoplasma mobile 163K and the nonfermentative strain Mycoplasma arthritidis ISR1. The addition of medium and the removal of culture suspension were regulated by acid production from glucose by M. mobile 163K and by ammonium production from arginine by M. arthritidis ISR1, respectively. For both strains the optimal pH for continuous growth was 7.0. The steady state could be maintained for at least 21 days. With CFU of 8.4 X 10(9) ml-1 (M. mobile 163K) and 3.2 X 10(9) ml-1 (M. arthritidis ISR1), the cell concentrations were slightly higher than those obtained in batch cultures. The dependence on the adjusted pH values was measured for several parameters, such as flow rate, CFU, glucose fermentation or production of ammonia, and gliding velocity. Since the long lag phases of batch cultures can be avoided, pH-controlled continuous cultures provide an appropriate system for the production of mycoplasma cells. PMID:2729987

  5. Antimycoplasmal Activity of Hydroxytyrosol

    Microsoft Academic Search

    Pio Maria Furneri; Anna Piperno; Antonella Sajia; Giuseppe Bisignano

    2004-01-01

    The aim of this study was to investigate the in vitro antimycoplasmal activity of hydroxytyrosol. Twenty strains of Mycoplasma hominis, three strains of Mycoplasma fermentans, and one strain of Mycoplasma pneu- moniae were used. For M. pneumoniae, M. hominis, and M. fermentans, the MICs were 0.5, 0.03 (for 90% of the strains tested), and 0.25 g\\/ml, respectively. Typical components of

  6. Characterization of hemadsorption-negative mutants of Mycoplasma pneumoniae.

    PubMed Central

    Hansen, E J; Wilson, R M; Clyde, W A; Baseman, J B

    1981-01-01

    Previously isolated mutants of Mycoplasma pneumoniae incapable of hemadsorption were characterized with respect to specific protein content, tracheal ring attachment capability, and virulence for both in vitro and in vivo model systems. Two-dimensional gel electrophoresis revealed both quantitative and qualitative differences between the protein complements of two different mutant strains and that of the virulent parent strain. Studies of mycoplasma attachment to hamster tracheal rings in vitro demonstrated that only one of these mutant strains still possessed the ability to attach to the respiratory epithelium via neuraminidase-sensitive receptors. Measurement of [3H]orotic acid uptake in mycoplasma-infected tracheal rings indicated that infection with the hemadsorption-negative mutants resulted in only slight reductions of ribonucleic acid synthesis, similar to levels observed for tracheal rings infected with an avirulent strain of M. pneumoniae. The virulence potential of the two mutant strains was further investigated by utilizing the hamster model system. Both mutant strains were rapidly cleared from the lungs of infected animals and produced little or no microscopic pneumonia. Images PMID:6163719

  7. [Identification of Placenta hominis and its adulterants using COI barcode].

    PubMed

    Chen, Jun; Jia, Jing; Xu, Xiao-Lan; Xin, Tian-Yi; Zhang, Hong-Yin; Shi, Lin-Chun; Yao, Hui; Liu, Dong; Wu, Zhen-Hong

    2014-06-01

    In order to provide a new method for the identification of Placenta hominis, the COI barcode has been employed to identify the P. hominis medicinal materials and its adulterants. Genomic DNA was extracted from the experimental samples. The COI sequences were amplified and sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. NJ tree was constructed by MEGA6.0 software. COI sequences can be successfully obtained from all experimental samples. The intra-specific variation and inter-specific divergence were calculated. The average intra-specific K2P distance of P. hominis was 0.001 and the maximum intra-specific distance was 0.008. The cluster dendrogram constructed can be seen that the same genus is together, and distinguished from its adulterants. It is concluded that P. hominis and its adulterants can be correctly identified by DNA barcoding method. PMID:25244745

  8. 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.

  9. [Membranoproliferative glomerulonephritis and mycoplasma infection].

    PubMed

    Dumas, R; Bascoul, S; Baldet, P; Serre, A; Roux, J; Jean, R

    1976-10-01

    Two children presented with an acute mycoplasma infection associated with a significant increase of antistreptolysin level. A severe nephropathy occurred, rapidly resulting in renal failure with histologic lesions of membrano-proliferative glomerulonephritis. Both patients had persisting low complement levels with low C3 and normal C4. The relationship between mycoplasma and streptococcal infections and the abnormality of complement and the renal disease is discussed. PMID:791180

  10. The Origin of the ‘Mycoplasma mycoides Cluster’ Coincides with Domestication of Ruminants

    PubMed Central

    Fischer, Anne; Shapiro, Beth; Muriuki, Cecilia; Heller, Martin; Schnee, Christiane; Bongcam-Rudloff, Erik; Vilei, Edy M.; Frey, Joachim; Jores, Joerg

    2012-01-01

    The ‘Mycoplasma mycoides cluster’ comprises the ruminant pathogens Mycoplasma mycoides subsp. mycoides the causative agent of contagious bovine pleuropneumonia (CBPP), Mycoplasma capricolum subsp. capripneumoniae the agent of contagious caprine pleuropneumonia (CCPP), Mycoplasma capricolum subsp. capricolum, Mycoplasma leachii and Mycoplasma mycoides subsp. capri. CBPP and CCPP are major livestock diseases and impact the agricultural sector especially in developing countries through reduced food-supply and international trade restrictions. In addition, these diseases are a threat to disease-free countries. We used a multilocus sequence typing (MLST) approach to gain insights into the demographic history of and phylogenetic relationships among the members of the ‘M. mycoides cluster’. We collected partial sequences from seven housekeeping genes representing a total of 3,816 base pairs from 118 strains within this cluster, and five strains isolated from wild Caprinae. Strikingly, the origin of the ‘M. mycoides cluster’ dates to about 10,000 years ago, suggesting that the establishment and spread of the cluster coincided with livestock domestication. In addition, we show that hybridization and recombination may be important factors in the evolutionary history of the cluster. PMID:22558362

  11. The origin of the 'Mycoplasma mycoides cluster' coincides with domestication of ruminants.

    PubMed

    Fischer, Anne; Shapiro, Beth; Muriuki, Cecilia; Heller, Martin; Schnee, Christiane; Bongcam-Rudloff, Erik; Vilei, Edy M; Frey, Joachim; Jores, Joerg

    2012-01-01

    The 'Mycoplasma mycoides cluster' comprises the ruminant pathogens Mycoplasma mycoides subsp. mycoides the causative agent of contagious bovine pleuropneumonia (CBPP), Mycoplasma capricolum subsp. capripneumoniae the agent of contagious caprine pleuropneumonia (CCPP), Mycoplasma capricolum subsp. capricolum, Mycoplasma leachii and Mycoplasma mycoides subsp. capri. CBPP and CCPP are major livestock diseases and impact the agricultural sector especially in developing countries through reduced food-supply and international trade restrictions. In addition, these diseases are a threat to disease-free countries. We used a multilocus sequence typing (MLST) approach to gain insights into the demographic history of and phylogenetic relationships among the members of the 'M. mycoides cluster'. We collected partial sequences from seven housekeeping genes representing a total of 3,816 base pairs from 118 strains within this cluster, and five strains isolated from wild Caprinae. Strikingly, the origin of the 'M. mycoides cluster' dates to about 10,000 years ago, suggesting that the establishment and spread of the cluster coincided with livestock domestication. In addition, we show that hybridization and recombination may be important factors in the evolutionary history of the cluster. PMID:22558362

  12. Systemic Disease in Vaal Rhebok (Pelea capreolus) Caused by Mycoplasmas in the Mycoides Cluster

    PubMed Central

    Nicolas, Melissa M.; Stalis, Ilse H.; Clippinger, Tracy L.; Busch, Martin; Nordhausen, Robert; Maalouf, Gabriel; Schrenzel, Mark D.

    2005-01-01

    In the winter of 2002, an outbreak of mycoplasma infection in Vaal rhebok (Pelea capreolus) originating from South Africa occurred 15 weeks after their arrival in San Diego, Calif. Three rhebok developed inappetence, weight loss, lethargy, signs related to pulmonary or arthral dysfunction, and sepsis. All three rhebok died or were euthanized. Primary postmortem findings were erosive tracheitis, pleuropneumonia, regional cellulitis, and necrotizing lymphadenitis. Mycoplasmas were detected in numerous tissues by electron microscopy, immunohistochemistry, and PCR. The three deceased rhebok were coinfected with ovine herpesvirus-2, and two animals additionally had a novel gammaherpesvirus. However, no lesions indicative of herpesvirus were seen microscopically in any animal. The rheboks' mycoplasmas were characterized at the level of the 16S rRNA gene, the 16S-23S intergenic spacer region, and the fructose biphosphate aldolase gene. Denaturing gradient gel electrophoresis was carried out to address the possibility of infection with multiple strains. Two of the deceased rhebok were infected with a single strain of Mycoplasma capricolum subsp. capricolum, and the third animal had a single, unique strain most closely related to Mycoplasma mycoides subsp. mycoides large-colony. A PCR survey of DNA samples from 46 other ruminant species demonstrated the presence of several species of mycoplasmas in the mycoides cluster, including a strain of M. capricolum subsp. capricolum identical to that found in two of the rhebok. These findings demonstrate the pervasiveness of mycoplasmas in the mycoides cluster in small ruminants and the potential for interspecies transmission and disease when different animal taxa come in contact. PMID:15750104

  13. 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.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  14. Increasing the value of hominy feed as a coproduct by fermentation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hominy feed is a low value ($83.7/metric ton) coproduct of the corn dry milling process that accounts for nearly 35% of the starting corn quantity. The average composition of hominy feed on a dry basis is 56.9% starch, 25.2% neutral detergent fiber, 11.1% protein, and 5.3% fat. Starch in hominy fe...

  15. Induced recurrent keratitis in rabbits infected with Herpesvirus hominis

    E-print Network

    Johnson, David Kendall

    1968-01-01

    evaluated for their capacity to induce re- tk tt kk't p ty' f td tk~H hominis. Agents tested were azathioprine, ultraviolet irradfation, and histam1ne phosphate. H. hominis was instilled into the cul-de- sac of the left eye of 40 rabbits and 36 responded.... Each agent was used on 8 animals. A recurrence rate of I of 8, 3 of 8, and 5 of 8 was observed for azathioprine, ultraviolet irrad1ation, and histamine phosphate, respectively. Virus isolates recovered from the reacting animals produced the same...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  17. Paenalcaligenes hominis gen. nov., sp. nov., a new member of the family Alcaligenaceae.

    PubMed

    Kämpfer, P; Falsen, E; Langer, S; Lodders, N; Busse, H-J

    2010-07-01

    A beige-pigmented bacterium (strain CCUG 53761A(T)) was isolated from human blood from an 85-year-old man in Göteborg, Sweden. Comparative analysis of 16S rRNA gene sequences showed that this bacterium displayed <95 % similarity to all described species of the genera of the family Alcaligenaceae. It grouped within the radiation of the genus Alcaligenes, but showed only 93.0-94.8 % similarity to type strains of members of this genus (Alcaligenes faecalis subsp. parafaecalis, 94.8 %; Alcaligenes faecalis subsp. faecalis, 94.2 %; Alcaligenes faecalis subsp. phenolicus, 93.4 %). This discrimination was supported by chemotaxonomic differences. The polyamine pattern consisted of the predominant compound putrescine, moderate amounts of spermidine and minor to trace amounts of spermine and cadaverine; 2-hydroxyputrescine was not detectable. The quinone system was ubiquinone Q-8 with minor amounts of Q-7. The polar lipid profile was composed of the major lipids diphosphatidylglycerol and phosphatidylethanolamine and moderate amounts of phosphatidylglycerol and an unknown phospholipid; minor lipids were also detected. The fatty acid profile, with large amounts of C(16 : 0) and C(17 : 0) cyclo and the absence of C(12 : 0) 2-OH as hydroxylated fatty acid, also differed significantly from those reported for Alcaligenes species. On the basis of these data, it is proposed that strain CCUG 53761A(T) represents a novel genus and species, for which the name Paenalcaligenes hominis gen. nov., sp. nov. is proposed. The type strain of Paenalcaligenes hominis is CCUG 53761A(T) =CCM 7698(T). PMID:19684310

  18. High-resolution melting-curve analysis of obg gene to differentiate the temperature-sensitive Mycoplasma synoviae vaccine strain MS-H from non-temperature-sensitive strains.

    PubMed

    Shahid, Muhammad A; Markham, Philip F; Marenda, Marc S; Agnew-Crumpton, Rebecca; Noormohammadi, Amir H

    2014-01-01

    Temperature-sensitive (ts+) vaccine strain MS-H is the only live attenuated M. synoviae vaccine commercially available for use in poultry. With increasing use of this vaccine to control M. synoviae infections, differentiation of MS-H from field M. synoviae strains and from rarely occurring non-temperature-sensitive (ts-) MS-H revertants has become important, especially in countries where local strains are indistinguishable from MS-H by sequence analysis of variable lipoprotein haemagglutinin (vlhA) gene. Single nucleotide polymorphisms (SNPs) in the obg of MS-H have been found to associate with ts phenotype. In this study, four PCRs followed by high-resolution melting (HRM)-curve analysis of the regions encompassing these SNPs were developed and evaluated for their potential to differentiate MS-H from 36 M. synoviae strains/isolates. The nested-obg PCR-HRM differentiated ts+ MS-H vaccine not only from field M. synoviae strains/isolates but also from ts- MS-H revertants. The mean genotype confidence percentages, 96.9±3.4 and 8.8±11.2 for ts+ and ts- strains, respectively, demonstrated high differentiating power of the nested-obg PCR-HRM. Using a combination of nested-obg and obg-F3R3 PCR-HRM, 97% of the isolates/strains were typed according to their ts phenotype with all MS-H isolates typed as MS-H. A set of respiratory swabs from MS-H vaccinated specific pathogen free chickens and M. synoviae infected commercial chicken flocks were tested using obg PCR-HRM system and results were consistent with those of vlhA genotyping. The PCR-HRM system developed in this study, proved to be a rapid and reliable tool using pure M. synoviae cultures as well as direct clinical specimens. PMID:24643035

  19. Comparative Biosynthesis of Ornithine and Lysine by Mycoplasma and L Forms

    PubMed Central

    Smith, Paul F.

    1966-01-01

    Smith, Paul F. (University of South Dakota, Vermillion). Comparative biosynthesis of ornithine and lysine by Mycoplasma and L forms. J. Bacteriol. 92:164–169. 1966.—Seven species of Mycoplasma, two L forms not requiring salt and their parent bacteria, and two yeasts were examined for enzymes involved in the biosynthesis of ornithine and lysine. All organisms tested, except two species of Mycoplasma and the yeasts, decarboxylated meso-?, ?-diaminopimelic acid. None of the Mycoplasma species or L forms was capable either of reducing ?-aminoadipic acid to its semialdehyde or of incorporating ?-aminoadipic acid-6-C14 into lysine. All organisms, except the yeasts and Mycoplasma sp. caprine strain 14, acetylated glutamic acid, and all organisms possessed N?-acetyl-l-ornithine:2-oxo-glutarate aminotransferase activity. N?-acetylornithase activity was negligible in all organisms except Proteus and its L form. No transacetylation between acetylglutamic acid and ornithine, and vice versa, was demonstrable in any of the organisms. Mycoplasma species appear to possess the bacterial pathway to lysine. Ornithine does not appear to arise from glutamic acid. PMID:5941275

  20. FURUNCULAR MYIASIS CAUSED BY DERMATOBIA HOMINIS IN A RETURNING TRAVELER

    PubMed Central

    Bhandari, Ramanath; Janos, David P.; Sinnis, Photini

    2007-01-01

    Furuncular myiasis caused by Dermatobia hominis is endemic throughout Central and South America. We report a case of furuncular myiasis in a traveler returned from Costa Rica. The case is unique because the primary care physician obtained magnetic resonance images. The images, however, do not show any characteristic features that assist in diagnosis. PMID:17360891

  1. Mycoplasma agassizii sp., nov., isolated from the upper respiratory tract of the desert tortoise (Gopherus agassizii) and the gopher tortoise (Gopherus polyphemus).

    USGS Publications Warehouse

    Brown, Mary E.; Brown, D.R.; Kelin, P.A.; McLaughlin, G.S.; Schumacher, I.M.; Jacobson, E.R.; Adams, H.P.; Tully, J.G.

    2001-01-01

    Biochemical, serological and molecular genetic studies were performed on seven mycoplasma isolates that were recovered from the upper respiratory tract of clinically ill desert tortoises. The isolates were serologically related to each other but serologically distinct from previously described species. Unique mycoplasma species-specific 16S rRNA nucleotide sequences were found in the proposed type strain. The name Mycoplasma agassizii is proposed for these isolates. The type strain is PS6T (=ATCC 700616T) which caused upper respiratory tract disease (URTD) in experimentally infected tortoises.

  2. [Mycoplasma mastitis in dairy cattle].

    PubMed

    Tolboom, R K; Snoep, J J; Sampimon, O C; Sol, J; Lam, T J G M

    2008-02-01

    The most important characteristics of Mycoplasma mastitis on dairy farms are described, based on two case studies. Clinical symptoms, diagnostics, epidemiology, and a plan of action are presented. In the herds investigated, Mycoplasma mastitis was characterized by multiple affected quarters unresponsive to treatment with antibiotic and/or anti-inflammatory agents. Most striking were a sandy sediment, brown colouring, and rice-like structure of the milk of affected animals. Clinical symptoms differed in the two affected herds. Diagnosis was based on bacteriological investigation of samples of milk and synovial fluid taken from infected cows. Affected animals were culled immediately, and the herds were monitored by repeated testing of bulk milk samples. It was concluded that a consequence of the increasing size of cattle herds in the Netherlands is that subclinical/clinical Mycoplasma mastitis may be diagnosed more frequently than in the past. In the case of Mycoplasma mastitis, farmers and veterinary practitioners are advised to draw up a plan of action together, incorporating aspects such as diagnostics at cow level, direct culling of affected animals, hygiene during milking, including post-milking teat disinfection, and routine monitoring of bulk milk. Unpasteurized milk should not be given to calves. PMID:18309823

  3. 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

  4. INDUCTION OF A MYCOPLASMA GALLISEPTICUM PMGA GENE IN THE CHICKEN TRACHEAL RING ORGAN CULTURE MODEL

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To search for Mycoplasma gallisepticum genes involved in colonization of the poultry respiratory tract, a transposon containing a promoterless lacZ gene was employed as a transcriptional reporter. The transposon was used to randomly mutagenize the chromosome of the M. gallisepticum S6 strain and a ...

  5. Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification.

    PubMed Central

    van Kuppeveld, F J; van der Logt, J T; Angulo, A F; van Zoest, M J; Quint, W G; Niesters, H G; Galama, J M; Melchers, W J

    1992-01-01

    Systematic computer alignment of mycoplasmal 16S rRNA sequences allowed the identification of variable regions with both genus- and species-specific sequences. Species-specific sequences of Mycoplasma collis were elucidated by asymmetric amplification and dideoxynucleotide sequencing of variable regions, using primers complementary to conserved regions of 16S rRNA. Primers selected for Mycoplasma pneumoniae, M. hominis, M. fermentans, Ureaplasma urealyticum, M. pulmonis, M. arthritidis, M. neurolyticum, M. muris, and M. collis proved to be species specific in the polymerase chain reaction. The genus-specific primers reacted with all mycoplasmal species investigated as well as with members of the genera Ureaplasma, Spiroplasma, and Acholeplasma. No cross-reaction was observed with members of the closely related genera Streptococcus, Lactobacillus, Bacillus, and Clostridium or with any other microorganism tested. On the basis of the high copy number of rRNA, a highly sensitive polymerase chain reaction assay was developed in which the nucleic acid content equivalent to a single organism could be detected. Images PMID:1381174

  6. Dermatobia hominis: Small Migrants Hidden in Your Skin.

    PubMed

    Zammarchi, Lorenzo; Viligiardi, Riccardo; Strohmeyer, Marianne; Bartoloni, Alessandro

    2014-10-01

    Myiasis is a parasitic infestation of vertebrate animal tissues due to maggots of two-winged flies (Diptera) that feed on living or necrotic tissue. Dermatobia hominis occurs widely in tropical parts of Latin America; it is the most common cause of furuncular myiasis in this region. The continuous increase in international travel has increased the possibility of observing this pathology outside endemic countries, especially in travelers returning from the tropics. If clinicians are aware of the possibility of the disease and its treatment options, this dermatosis can be easily managed. However, diagnostic delay is very common because the disease is often misdiagnosed as a bacterial skin infection. Here, we report 2 cases of furuncular myiasis caused by D. hominis in travelers returning to Italy from Latin America. Surgical and noninvasive treatment approaches are also described. PMID:25324659

  7. The importance of B-cells and ecto-5?nucleotidase in Mycoplasma fermentans infection and the relevance to rheumatoid arthritis

    PubMed Central

    Johnson, Sheena M

    2008-01-01

    The aim of this work was to discover if Mycoplasma fermentans, which is known to infect B cells, could be the cause of the raised ecto-5?-nucleotidase observed in the synovial fluid of rheumatoid arthritis patients. The ecto-5?-nucleotidase activity in the patients' serum has been shown to correlate with the erythrocyte sedimentation rate and DNA from the mycoplasma has been found in the synovial fluid. B lymphoblastoid cell lines were exposed to 16 strains of Mycoplasma fermentans and their ecto-5?-nucleotidase, CD73, was measured both biochemically and by mouse antibodies to human ecto 5?-nucleotidase using the fluorescence activated cell sorter. The type strain, PG 18, did not grow with the B cells. Some of the mycoplasma strains (9/15) increased the cellular ecto-5?-nucleotidase activity from twice to 17 fold, and usually showed 5?-nucleotidase activity themselves. At least one strain, M106, induced human 5?-nucleotidase on the normally 5?-nucleotidase negative Daudi and Raji Burkitt's lymphoma cell lines, and increased sevenfold the 5?-nucleotidase on the monocyte/macrophage cell line THP-1. Growing the cells in aged medium increased the level of mycoplasma infection. This mycoplasma-induced enzyme showed a conformational change and an increase in activity with a glycosylation change involving mannose groups. The other group of strains, mostly of respiratory or cell culture origin, usually did not have any 5?-nucleotidase of their own and decreased the B-cell enzyme activity by about half. Electron microscopy and flow cytometry showed that the strain M106 was filamentous and could be found inside the B-cells. The 5?-nucleotidase-inducing strains of M. fermentans may be important in the aetiology of rheumatoid arthritis. PMID:17680797

  8. Highly Dynamic Genomic Loci Drive the Synthesis of Two Types of Capsular or Secreted Polysaccharides within the Mycoplasma mycoides Cluster

    PubMed Central

    Bertin, Clothilde; Pau-Roblot, Corinne; Courtois, Josiane; Manso-Silván, Lucía; Tardy, Florence; Poumarat, François; Citti, Christine; Sirand-Pugnet, Pascal; Gaurivaud, Patrice

    2014-01-01

    Mycoplasmas of the Mycoplasma mycoides cluster are all ruminant pathogens. Mycoplasma mycoides subsp. mycoides is responsible for contagious bovine pleuropneumonia and is known to produce capsular polysaccharide (CPS) and exopolysaccharide (EPS). Previous studies have strongly suggested a role for Mycoplasma mycoides subsp. mycoides polysaccharides in pathogenicity. Mycoplasma mycoides subsp. mycoides-secreted EPS was recently characterized as a ?(1?6)-galactofuranose homopolymer (galactan) identical to the capsular product. Here, we extended the characterization of secreted polysaccharides to all other members of the M. mycoides cluster: M. capricolum subsp. capripneumoniae, M. capricolum subsp. capricolum, M. leachii, and M. mycoides subsp. capri (including the LC and Capri serovars). Extracted EPS was characterized by nuclear magnetic resonance, resulting in the identification of a homopolymer of ?(1?2)-glucopyranose (glucan) in M. capricolum subsp. capripneumoniae and M. leachii. Monoclonal antibodies specific for this glucan and for the Mycoplasma mycoides subsp. mycoides-secreted galactan were used to detect the two polysaccharides. While M. mycoides subsp. capri strains of serovar LC produced only capsular galactan, no polysaccharide could be detected in strains of serovar Capri. All strains of M. capricolum subsp. capripneumoniae and M. leachii produced glucan CPS and EPS, whereas glucan production and localization varied among M. capricolum subsp. capricolum strains. Genes associated with polysaccharide synthesis and forming a biosynthetic pathway were predicted in all cluster members. These genes were organized in clusters within two loci representing genetic variability hot spots. Phylogenetic analysis showed that some of these genes, notably galE and glf, were acquired via horizontal gene transfer. These findings call for a reassessment of the specificity of the serological tests based on mycoplasma polysaccharides. PMID:25398856

  9. Efficacy of abamectin injection against Dermatobia hominis in cattle

    Microsoft Academic Search

    J. B. Cruz; C. Benitez-Usher; L. G. Cramer; S. J. Gross; A. B. Kohn

    1993-01-01

    The efficacy of abamectin 1%, when injected subcutancously in cattle at a dose of 200 µg\\/kg body weight, against the larval stages (grubs) of the flyDermatobia hominis was evaluated in two trials in endemic areas of Brazil and Argentina. Eighteen Holstein x Brahman castrated males and 16 Brahman-cross with natural infestations were used. Larvae were counted by instar in situ

  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 able to stimulate as well as suppress lymphocytes in a nonspecific, polyclonal manner, both in vitro and in vivo. As well as to affecting various subsets of lymphocytes, mycoplasmas and mycoplasma-derived cell components modulate the activities of monocytes/macrophages and NK cells and trigger the production of a wide variety of up-regulating and down-regulating cytokines and chemokines. Mycoplasma-mediated secretion of proinflammatory cytokines, such as tumor necrosis factor alpha, interleukin-1 (IL-1), and IL-6, by macrophages and of up-regulating cytokines by mitogenically stimulated lymphocytes plays a major role in mycoplasma-induced immune system modulation and inflammatory responses. PMID:9841667

  11. Absence of Mycoplasma Contamination in the Anthrax Vaccine

    PubMed Central

    Hart, Mary Kate; Del Giudice, Richard A.

    2002-01-01

    Mycoplasma contamination of the licensed anthrax vaccine administered to military personnel has been suggested as a possible cause of Persian Gulf illness. Vaccine samples tested by nonmilitary laboratories were negative for viable mycoplasma and mycoplasma DNA and did not support its survival. Mycoplasma contamination of anthrax vaccine should not be considered a possible cause of illness. PMID:11840996

  12. Original article In vivo transmission studies of `Candidatus Mycoplasma

    E-print Network

    Paris-Sud XI, Université de

    of the three feline haemotropic mycoplasmas ­ Mycoplasma haemofelis, `Candidatus Mycoplasma haemominutum of the feline haemotropic mycoplasma isolatesstudied todate varies, and immunosuppression or pre-existing retroviral infections may potentiate the severity of the anaemia [9, 36]. The diagnosis of feline haemotropic

  13. Cytotoxicity of Mycoplasma mycoides subsp. mycoides Small Colony Type to Bovine Epithelial Cells?

    PubMed Central

    Bischof, Daniela F.; Janis, Carole; Vilei, Edy M.; Bertoni, Giuseppe; Frey, Joachim

    2008-01-01

    The cytotoxicities of various strains of Mycoplasma mycoides subsp. mycoides small colony type (SC), the agent of contagious bovine pleuropneumonia (CBPP), were measured in vitro using embryonic calf nasal epithelial (ECaNEp) cells. Strains isolated from acute cases of CBPP induced high cytotoxicity in the presence of glycerol, concomitant with the release of large amounts of toxic H2O2 that were found to be translocated into the cytoplasms of the host cells by close contact of the Mycoplasma strains with the host cells. Currently used vaccine strains also showed high cytotoxicity and high H2O2 release, indicating that they are attenuated in another virulence attribute. Strains isolated from recent European outbreaks of CBPP with mild clinical signs, which are characterized by a defect in the glycerol uptake system, released small amounts of H2O2 and showed low cytotoxicity to ECaNEp cells. M. mycoides subsp. mycoides SC strain PG1 released large amounts of H2O2 but was only slightly cytotoxic. PG1 was found to have a reduced capacity to bind to ECaNEp cells and was unable to translocate H2O2 into the bovine cells, in contrast to virulent strains that release large amounts of H2O2. Thus, an efficient translocation of H2O2 into host cells is a prerequisite for the cytotoxic effect and requires an intact adhesion mechanism to ensure a close contact between mycoplasmas and host cells. PMID:17998309

  14. Transcriptome Changes in Mycoplasma hyopneumoniae during Infection

    Microsoft Academic Search

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

    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

  15. Recent Advances in Mycoplasma gallisepticum Vaccine Administration

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Application of live Mycoplasma gallisepticum vaccines to layer chickens generally occurs at 9 to 10 weeks of age. Mycoplasma organisms are extremely fastidious in the laboratory and difficult to grow. Very little attention has been accorded to optimizing parameters for vaccine administration in th...

  16. Lipoprotein Multigene Families in Mycoplasma pneumoniae

    Microsoft Academic Search

    K. M. Hallamaa; G. F. Browning; S. L. Tang

    2006-01-01

    There are several mechanisms used by pathogenic bacteria to evade host defenses, including antigenic variation on their surfaces. Antigenic variation of cell surface lipoproteins has been reported in a number of Mycoplasma species (3). Sequencing of the Mycoplasma pneumoniae genome has re- vealed that it harbors 46 lipoprotein genes in six multigene families, all of unknown function (4, 7). These

  17. Inferences about the Global Population Structures of Cryptosporidium parvum and Cryptosporidium hominis

    Microsoft Academic Search

    S. Tanriverdi; Alex Grinberg; Rachel M. Chalmers; Paul R. Hunter; Zorana Petrovic; Donna E. Akiyoshi; Eric London; Linghui Zhang; Saul Tzipori; James K. Tumwine; Giovanni Widmer

    2008-01-01

    Cryptosporidium parvum and Cryptosporidium hominis are two related species of apicomplexan protozoa responsible for the majority of human cases of cryptosporidiosis. In spite of their considerable public health impact, little is known about the population structures of these species. In this study, a battery of C. parvum and C. hominis isolates from seven countries was genotyped using a nine-locus DNA

  18. Cyclospora papionis, Cryptosporidium hominis, and human-pathogenic Enterocytozoon bieneusi in captive baboons in Kenya.

    PubMed

    Li, Wei; Kiulia, Nicholas M; Mwenda, Jason M; Nyachieo, Atunga; Taylor, Maureen B; Zhang, Xichen; Xiao, Lihua

    2011-12-01

    Cyclospora papionis, Cryptosporidium hominis, and Enterocytozoon bieneusi were detected in 42 (17.9%), 6 (2.6%), and 29 (12.3%) of 235 newly captured baboons in Kenya, respectively. Most C. hominis subtypes and E. bieneusi genotypes found have been detected in humans in the area, suggesting that cross-species transmission of cryptosporidiosis and microsporidiosis is possible. PMID:21956988

  19. Cyclospora papionis, Cryptosporidium hominis, and Human-Pathogenic Enterocytozoon bieneusi in Captive Baboons in Kenya?

    PubMed Central

    Li, Wei; Kiulia, Nicholas M.; Mwenda, Jason M.; Nyachieo, Atunga; Taylor, Maureen B.; Zhang, Xichen; Xiao, Lihua

    2011-01-01

    Cyclospora papionis, Cryptosporidium hominis, and Enterocytozoon bieneusi were detected in 42 (17.9%), 6 (2.6%), and 29 (12.3%) of 235 newly captured baboons in Kenya, respectively. Most C. hominis subtypes and E. bieneusi genotypes found have been detected in humans in the area, suggesting that cross-species transmission of cryptosporidiosis and microsporidiosis is possible. PMID:21956988

  20. Sedimentation Counting and Morphology of Mycoplasma

    PubMed Central

    Clark, Harold W.

    1965-01-01

    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 OsO4 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 m?) containing a limited deoxyribonucleic acid genetic coding molecule (approximately 4 × 106). Images PMID:5321487

  1. Diagnosis of contagious caprine pleuropneumonia by detection and identification of Mycoplasma capricolum subsp. capripneumoniae by PCR and restriction enzyme analysis.

    PubMed

    Bölske, G; Mattsson, J G; Bascuñana, C R; Bergström, K; Wesonga, H; Johansson, K E

    1996-04-01

    Contagious caprine pleuropneumonia (CCPP), one of the most serious and dramatic diseases of goats, is caused by Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae). This organism is very difficult to isolate and to correctly identify. In a previous report we described a method for the rapid detection and identification of M. capripneumoniae. This method is based on a PCR system by which a segment of the 16S rRNA gene from all mycoplasmas of the M. mycoides cluster can be amplified. The PCR product is then analyzed by restriction enzyme cleavage for the identification of M. capripneumoniae DNA. This system has now been further evaluated with respect to specificity and diagnostic efficacy for the identification and direct detection of the organism in clinical material. Identification by restriction enzyme analysis of amplified DNA from mycoplasmas of the M. mycoides cluster was verified for 55 strains, among which were 15 strains of M. capripneumoniae. The PCR was applied to clinical samples from the nose, ear, pharynx, pleural fluid, and lung tissue containing M. capripneumoniae or other mycoplasmas. As expected, mycoplasmas belonging to the M. mycoides cluster could be detected by the PCR. Restriction enzyme analysis of the PCR products could then be applied for the identification of M. capripneumoniae. Clinical samples and cultures containing M. capripneumoniae were dried on filter paper, to try an easier sample transport method, and were tested by PCR. M. capripneumoniae DNA could be detected in the dried specimens, but the sensitivity of the PCR test was reduced. PMID:8815084

  2. Insights into the function of Mycoplasma pneumoniae protein P30 from orthologous gene replacement.

    PubMed

    Relich, Ryan F; Balish, Mitchell F

    2011-10-01

    The attachment organelles of bacterial species belonging to the Mycoplasma pneumoniae phylogenetic cluster are required for host cytadherence, gliding motility and virulence. Despite being closely related, these bacteria possess distinct cellular morphologies and gliding characteristics. The molecular mechanisms for most attachment organelle phenotypes, including shape and ability to power motility, are obscure. The attachment organelle-associated P30 protein of M. pneumoniae is implicated in both adherence and motility, with mutations negatively impacting cell morphology, adherence, gliding and virulence. To test whether the P30 alleles of different mycoplasma species confer species-specific attachment organelle properties, we created an M. pneumoniae strain in which the Mycoplasma genitalium P30 orthologue, P32, was substituted for the native P30. Selected clones were visualized by scanning electron microscopy to assess morphology and by indirect immunofluorescence microscopy to localize P32. Cytadherence ability and gliding motility were assessed by haemadsorption assay and phase-contrast microcinematography, respectively. Cell and attachment organelle morphologies were indistinguishable from wild-type M. pneumoniae as well as M. pneumoniae II-3 expressing a C-terminally 6×His-tagged P30 construct. P32 was localized to the tip of the attachment organelle of transformant cells. Although a specific role for P30 in species-specific phenotypes was not identified, this first test of orthologous gene replacement in different mycoplasma species demonstrates that the differences in the M. pneumoniae and M. genitalium proteins contribute little if anything to the different attachment organelle phenotypes between these species. PMID:21778204

  3. Insights into the function of Mycoplasma pneumoniae protein P30 from orthologous gene replacement

    PubMed Central

    Relich, Ryan F.

    2011-01-01

    The attachment organelles of bacterial species belonging to the Mycoplasma pneumoniae phylogenetic cluster are required for host cytadherence, gliding motility and virulence. Despite being closely related, these bacteria possess distinct cellular morphologies and gliding characteristics. The molecular mechanisms for most attachment organelle phenotypes, including shape and ability to power motility, are obscure. The attachment organelle-associated P30 protein of M. pneumoniae is implicated in both adherence and motility, with mutations negatively impacting cell morphology, adherence, gliding and virulence. To test whether the P30 alleles of different mycoplasma species confer species-specific attachment organelle properties, we created an M. pneumoniae strain in which the Mycoplasma genitalium P30 orthologue, P32, was substituted for the native P30. Selected clones were visualized by scanning electron microscopy to assess morphology and by indirect immunofluorescence microscopy to localize P32. Cytadherence ability and gliding motility were assessed by haemadsorption assay and phase-contrast microcinematography, respectively. Cell and attachment organelle morphologies were indistinguishable from wild-type M. pneumoniae as well as M. pneumoniae II-3 expressing a C-terminally 6×His-tagged P30 construct. P32 was localized to the tip of the attachment organelle of transformant cells. Although a specific role for P30 in species-specific phenotypes was not identified, this first test of orthologous gene replacement in different mycoplasma species demonstrates that the differences in the M. pneumoniae and M. genitalium proteins contribute little if anything to the different attachment organelle phenotypes between these species. PMID:21778204

  4. In vitro spatial and temporal analysis of Mycoplasma pneumoniae colonization of human airway epithelium.

    PubMed

    Prince, Oliver A; Krunkosky, Thomas M; Krause, Duncan C

    2014-02-01

    Mycoplasma pneumoniae is an important cause of respiratory disease, especially in school-age children and young adults. We employed normal human bronchial epithelial (NHBE) cells in air-liquid interface culture to study the interaction of M. pneumoniae with differentiated airway epithelium. These airway cells, when grown in air-liquid interface culture, polarize, form tight junctions, produce mucus, and develop ciliary function. We examined both qualitatively and quantitatively the role of mycoplasma gliding motility in the colonization pattern of developing airway cells, comparing wild-type M. pneumoniae and mutants thereof with moderate to severe defects in gliding motility. Adherence assays with radiolabeled mycoplasmas demonstrated a dramatic reduction in binding for all strains with airway cell polarization, independent of acquisition of mucociliary function. Adherence levels dropped further once NHBE cells achieved terminal differentiation, with mucociliary activity strongly selecting for full gliding competence. Analysis over time by confocal microscopy demonstrated a distinct colonization pattern that appeared to originate primarily with ciliated cells, but lateral spread from the base of the cilia was slower than expected. The data support a model in which the mucociliary apparatus impairs colonization yet cilia provide a conduit for mycoplasma access to the host cell surface and suggest acquisition of a barrier function, perhaps associated with tethered mucin levels, with NHBE cell polarization. PMID:24478073

  5. Persistence of Mycoplasma gallisepticum in chickens after treatment with enrofloxacin without development of resistance.

    PubMed

    Reinhardt, Anita K; Gautier-Bouchardon, Anne V; Gicquel-Bruneau, Mireille; Kobisch, Marylène; Kempf, Isabelle

    2005-03-20

    The ability of the avian pathogen Mycoplasma gallisepticum to persist despite fluoroquinolone treatment was investigated in chickens. Groups of specific pathogen free chickens were experimentally infected with M. gallisepticum and treated with enrofloxacin at increasing concentrations up to the therapeutic dose. When M. gallisepticum could no longer be re-isolated from chickens, birds were stressed by inoculation of infectious bronchitis virus or avian pneumovirus. Although M. gallisepticum could not be cultured from tracheal swabs collected on several consecutive sampling days after the end of the enrofloxacin treatments, the infection was not eradicated. Viral infections reactivated the mycoplasma infection. Mycoplasmas were isolated from tracheal rings cultured for several days, suggesting that M. gallisepticum persisted in the trachea despite the enrofloxacin treatment. The minimal inhibitory concentration (MIC) of enrofloxacin for most of the re-isolated mycoplasmas was the same as that of the strain with which the birds were inoculated. Furthermore, no mutation could be detected in the fluoroquinolone target genes. These results suggest that M. gallisepticum can persist in chickens without development of resistance despite several treatments with enrofloxacin. PMID:15737482

  6. Mycoplasmas and Oncogenesis: Persistent Infection and Multistage Malignant Transformation

    Microsoft Academic Search

    Shien Tsai; Douglas J. Wear; James Wai-Kuo Shih; Shyh-Ching Lo

    1995-01-01

    Oncogenic potential of human mycoplasmas was studied using cultured mouse embryo cells, C_3H\\/10T1\\/2 (C_3H). Mycoplasma fermentans and Mycoplasma penetrans, mycoplasmas found in unusually high frequencies among patients with AIDS, were examined. Instead of acute transformation, a multistage process in promotion and progression of malignant cell transformation with long latency was noted; after 6 passages (1 wk per passage) of persistent

  7. The complete genome sequence of the murine respiratory pathogen Mycoplasma pulmonis

    Microsoft Academic Search

    Isabelle Chambaud; Roland Heilig; Stéphane Ferris; Valérie Barbe; Delphine Samson; Frédérique Galisson; I van Moszer; Kevin Dybvig; Henri Wróblewski; Alain Viari; Eduardo P. C. Rocha; Alain Blanchard

    2001-01-01

    Mycoplasma pulmonis is a wall-less eubacterium belonging to the Mollicutes (trivial name, myco- plasmas) and responsible for murine respiratory diseases. The genome of strain UAB CTIP is composed of a single circular 963 879 bp chromosome with a G + C content of 26.6 mol%, i.e. the lowest reported among bacteria, Ureaplasma urealyticum apart. This genome contains 782 putative coding

  8. The complete genomic sequence of Mycoplasma penetrans, an intracellular bacterial pathogen in humans

    Microsoft Academic Search

    Yuko Sasaki; Jun Ishikawa; Atsushi Yamashita; Kenshiro Oshima; Tsuyoshi Kenri; Keiko Furuya; Chie Yoshino; Atsuko Horino; Tadayoshi Shiba; Tsuguo Sasaki; Masahira Hattori

    2002-01-01

    The complete genomic sequence of an intracellular bacterial pathogen, Mycoplasma penetrans HF-2 strain, was determined. The HF-2 genome consists of a 1 358 633 bp single circular chromosome con- taining 1038 predicted coding sequences (CDSs), one set of rRNA genes and 30 tRNA genes. Among the 1038 CDSs, 264 predicted proteins are common to the Mycoplasmataceae sequenced thus far and

  9. Inactivation of mycoplasma in serum with binary ethyleneimine.

    PubMed Central

    Christianson, G G; Pemberton, J R; Koski, T A

    1980-01-01

    A method of inactivating mycoplasma contaminants in sera by using binary ethyleneimine was tried. When used at the concentration recommended for inactivating viruses, 0.001 M, binary ethyleneimine inactivated only two of five mycoplasma species. Even at a concentration of 0.01 M, the compound did not consistently inactivate all mycoplasmas. PMID:7372800

  10. Passive immunization in experimental Herpesvirus hominis infection of newborn mice.

    PubMed Central

    Luyet, F; Samra, D; Soneji, A; Marks, M I

    1975-01-01

    Infection of newborn mice with Herpesvirus hominis type 2(HVH-2) was used as an experimental model of disseminated HVH infection in newborn humans. Mice were challenged with 103 plaque-forming units of HVH-2 intranasally and were given 0.2 ml of rabbit serum intraperitoneally. Passive immunizations with rabbit anti-HVH-2 serum resulted in a significant decrease in mortality and prolongation of survival time. This effect correlated with the neutralizing antibody titer of the serum against HVH-2 and was more pronounced when immune serum was administered 1 h after infection as compared with 24 h. These results suggest that administration of high-titer anti-HVH-2 immunoglobulins shortly after delivery could afford significant protection to the newborn of a mother with genital HVH-2 infection. PMID:173653

  11. Comparison of isolates of Mycoplasma mycoides subspecies capri from asymptomatic and septicaemic goats.

    PubMed

    Tardy, F; Maigre, L; Tricot, A; Poumarat, F; Nguyen, L; Le Grand, D

    2011-01-01

    Strains of Mycoplasma mycoides subspecies capri (Mmc) are frequently isolated from goats with contagious agalactia, but they can also be recovered from herds that have shown no clinical signs of mycoplasmosis for several years. The present study was conducted in order to explore the potential genetic and antigenic differences existing between an Mmc strain isolated from an outbreak (septicaemic) and a strain isolated from the ear canal of a goat belonging to a herd with no recent episode of mycoplasmosis (carriage strain). The genomes of the two strains, compared by suppression subtractive hybridization, were shown to be poorly divergent. The two strains were inoculated into goats to produce specific antisera, but both induced fatal mycoplasmosis. These results indicate that septicaemic and carriage strains cannot be distinguished by their genetic background or by their pathogenic capacity under experimental conditions. PMID:20708197

  12. [The case of Henoch-Schönlein Purpura associated with Blastocystis hominis].

    PubMed

    Tutanç, Murat; Silfeler, Ibrahim; Ozgür, Tümay; Motor, Vicdan Köksald?; Kurto?lu, Ahmet Ibrahim

    2013-01-01

    Blastocystis hominis (B. hominis) is a parasite that often causes gastrointestinal symptoms in patients with immune deficiency and has a controversial pathogenicity in healthy people, although some symptoms are reported outside of the gastrointestinal system in healthy persons. Henoch-Schönlein Purpura (HSP) vasculitis is an acute autoimmune disease characterised by IgA storage of small vessels that is believed to include infectious factors in its aetiology. A 30-month follow-up with a boy diagnosed with HSP being treated with steroid therapy showed that he had recurrent symptoms within two days, and B. hominis was detected in the faecal analysis. His symptoms including rash, abdominal pain, and arthritis improved after treatment with steroid and co-trimaksazol. This paper is the first to present a case of HSP associated with B. hominis. PMID:23955912

  13. Blastocystis hominis and Dientamoeba fragilis in patients fulfilling irritable bowel syndrome criteria.

    PubMed

    Yakoob, Javed; Jafri, Wasim; Beg, Mohammad Asim; Abbas, Zaigham; Naz, Shagufta; Islam, Muhammad; Khan, Rustam

    2010-08-01

    Studies have suggested a possible role for Blastocystis hominis and Dientamoeba fragilis in the etiology of irritable bowel syndrome (IBS). We studied the prevalence of B. hominis and D. fragilis in patients with IBS-diarrhea (IBS-D). Three hundred and thirty patients were enrolled, 171 (52%) with IBS-D and 159 (48%) were controls, respectively. Stool microscopy, culture, and polymerase chain reaction (PCR) for B. hominis and D. fragilis were done. B. hominis was positive by stool microscopy in 49% (83/171) of IBS compared to 24% (27/159) in control (p < 0.001). B. hominis culture was positive in 53% (90/171) in IBS compared to 16% (25/159) in control (p < 0.001). B. hominis PCR was positive in 44% (75/171) in IBS compared to 21% (33/159) in control (p < 0.001). D. fragilis microscopy was positive in 3.5% (6/171) in IBS-D compared to 0.6% (1/159) in control (p = 0.123). D. fragilis culture was positive in 4% (7/171) in IBS compared to 1.3% (2/159) in control (p = 0.176). D. fragilis PCR was positive in 4% (6/171) in IBS-D compared to 0% (0/159) in control (p = 0.030). B. hominis is common, while D. fragilis was less prevalent in our patients with IBS-D. B. hominis and D. fragilis culture had a better yield compared to stool microscopy and PCR. PMID:20532564

  14. Stress Exacerbates Infectivity and Pathogenicity of Blastocystis hominis: In Vitro and In Vivo Evidences

    PubMed Central

    Chandramathi, Samudi; Suresh, Kumar; Sivanandam, Sinnadurai; Kuppusamy, Umah Rani

    2014-01-01

    Background Stress alters the oxidant-antioxidant state and immune cell responses which disrupts its function to combat infection. Blastocystis hominis, a common intestinal protozoan has been reported to be opportunistic in immunocompromised patients namely cancer. B. hominis infectivity in other altered immune system conditions especially stress is unknown. We aimed to demonstrate the stress effects towards the susceptibility and pathogenicity of B. hominis infection. Methods/Findings Three-week-old Wistar rats were divided into four groups: (a)control; (b)stress-induced; (c)B. hominis infected; (d)stress-induced with B. hominis infection; (n?=?20 respectively). Stress was induced for an hour daily (30 days) using a Belly Dancer Shaker. Weight gain was monitored, stool samples were collected for B. hominis screening and blood for the determination of differential count, levels of immunoglobulin, oxidative damage, and peripheral blood mononuclear cell (PBMC) proliferation upon induction with solubilized antigen of B. hominis (Blasto-Ag). Group (b) exhibited the highest level of weight gain. Group (d) had higher levels of parasite cyst count in stools, serum IgE, oxidized protein and lipid compared to the group (c). Levels of monocyte and antioxidant in group (d) were decreased and their PBMCs showed highest inhibition of proliferation level when exposed to Blasto-Ag. Monocyte level in Group (b) showed insignificant difference compared to group (a) but was significantly lower compared to group (c). Antioxidant levels in group (c) were generally lower compared to group (a) and (b). Inhibition level exhibited by Blasto-Ag treated PBMCs of group (c) was higher compared to group (a) and (b). Conclusion The pathogenicity and augmentation of B. hominis infection is enhanced when stress is present. Lifestyles today are becoming increasingly stressed and the present findings suggest that the parasite which has been reported to be one of the most common organisms seen in stool surveys, namely in developing countries, may tend to be more pathogenic in stressful situations. PMID:24788756

  15. Stimulation of mouse lymphocytes by a mitogen derived from Mycoplasma arthritidis (MAM). VIII. Selective activation of T cells expressing distinct V beta T cell receptors from various strains of mice by the "superantigen" MAM.

    PubMed

    Cole, B C; Kartchner, D R; Wells, D J

    1990-01-15

    Mycoplasma arthritidis T cell mitogen (MAM), in association with its MHC ligand, is recognized by T cells that express TCR-alpha/beta assembled with a product(s) of the V beta 8 gene family. We show here that lymphocytes from mice which fail to express V beta 8 products can also be activated by MAM and the resulting cultures exhibit a marked increase in V beta 6 TCR-bearing cells. Evidence was also obtained that MAM can activate T cells that express all three V beta 8 TCR. The mAb, F23.1, which recognizes all V beta 8 gene products, was strongly inhibitory for MAM-induced proliferation of CBA cells whose T cell repertoire for MAM consists of T cells that express V beta 8.2 and 8.3 TCR. In contrast, the F23.1 mAb was only weakly inhibitory for BALB/c splenocytes which express V beta 6 TCR in addition to all three V beta 8 TCR. Involvement of V beta 8.1, 8.2, 8.3, and V beta 6 in MAM-induced proliferation was confirmed by expanding lymphocyte cultures in the presence of MAM and phenotyping the activated cells for expression of individual V beta TCR. There was also evidence for a selective activation of T cells bearing specific V beta TCR because BALB/c T cell populations expanded with MAM were comprised of 46.2% V beta 8.2+ cells, 18.6% V beta 8.1+ cells, 7.6% V beta 8.3+ cells and 6.7% V beta 6+ cells. Recent studies suggest that the newly described "superantigens" including the staphylococcal enterotoxins and the self minor lymphocyte-stimulating Ag activate T cells in a manner similar to that described earlier for MAM. The discovery of shared recognition of these proteins by specific V beta TCR strongly suggests that MAM belongs to the superantigen protein family, the members of which may share cross-reactive epitopes. Inasmuch as MAM is produced by an organism which induces chronic joint disease, our findings provide the basis for a new model to study the role of superantigens in the development of chronic autoimmune type diseases. PMID:2136890

  16. A study on Blastocystis hominis in food-handlers: diagnosis and potential pathogenicity.

    PubMed

    Fathy, Fouad M

    2011-08-01

    Proper diagnosis of Blastocystis hominis in not performed routinely in medical laboratories of developing countries; consequently clinical significance of this common intestinal protozoon is liable to remain unsettled. Food-handlers are more prone to get and transmit this feco-oral infection. This work compared the sensitivity of direct diagnostic methods to detect B. hominis in stool, estimate the true prevalence among food-handlers in Sirte-Libya, to clarify the association between the parasite and gastrointestinal symptoms and the response to specific treatment. A total of 400 male food-handlers aged 18-50 year were included. Each was subjected to clinical questionnaire and 3 stool examinations by different methods. The results showed high prevalence of B. hominis in food-handlers (35.5%). Short- term in vitro culture (on Boeck and Derbholav's medium) was the most sensitive method for detection of B. hominis (35.5%), followed by permanent Trichrome-stained smear (27.5%); saline-sedimentation concentrated smear (21%) and direct iodine smear (14%). Of 108 cases having B. hominis alone, 68.5% were symptomatic. Diarrhea was the most frequent symptom (75.6%), followed by abdominal pain (66.2%) and flatulence (43.2%). Fecal parasite-load was significantly higher in symptomatic cases than asymptomatic; parasite and symptoms disappeared after metronidazole treatment. So, culture should be used on routine basis to detect B. hominis which should be considered pathogenic particularly when present alone in large numbers in symptomatic patients. PMID:21980782

  17. Comparative susceptibilities of various animal-pathogenic mycoplasmas to fluoroquinolones.

    PubMed Central

    Hannan, P C; Windsor, G D; de Jong, A; Schmeer, N; Stegemann, M

    1997-01-01

    The in vitro activities of six antimicrobial agents were tested against 162 mycoplasma strains of eight species isolated from poultry and livestock at different geographic sites. Tiamulin was most active (MICs at which 90% of the isolates were inhibited [MIC90s], 0.025 to 0.25 microg/ml); enrofloxacin and danofloxacin had near equivalent activities (MIC90s, 0.05 to 1.0 microg/ml), but were much more active than flumequine (MIC90s, 1 to 50 microg/ml). The MIC90s of tylosin and oxytetracycline were 0.25 to > 100 microg/ml and 0.25 to 100 microg/ml, respectively. PMID:9303412

  18. Genetic diversity of Mycoplasma hyopneumoniae isolates of abattoir pigs.

    PubMed

    Charlebois, Audrey; Marois-Créhan, Corinne; Hélie, Pierre; Gagnon, Carl A; Gottschalk, Marcelo; Archambault, Marie

    2014-01-31

    Mycoplasma hyopneumoniae, the causative agent of porcine enzootic pneumonia, is present in swine herds worldwide. However, there is little information on strains infecting herds in Canada. A total of 160 swine lungs with lesions suggestive of enzootic pneumonia originating from 48 different farms were recovered from two slaughterhouses and submitted for gross pathology. The pneumonic lesion scores ranged from 2% to 84%. Eighty nine percent of the lungs (143/160) were positive for M. hyopneumoniae by real-time PCR whereas 10% (16/160) and 8.8% (14/160) were positive by PCR for M. hyorhinis and M. flocculare, respectively. By culture, only 6% of the samples were positive for M. hyopneumoniae (10/160). Among the selected M. hyopneumoniae-positive lungs (n=25), 9 lungs were co-infected with M. hyorhinis, 9 lungs with PCV2, 2 lungs with PRRSV, 12 lungs with S. suis and 10 lungs with P. multocida. MLVA and PCR-RFLP clustering of M. hyopneumoniae revealed that analyzed strains were distributed among three and five clusters respectively, regardless of severity of lesions, indicating that no cluster is associated with virulence. However, strains missing a specific MLVA locus showed significantly less severe lesions and lower numbers of bacteria. MLVA and PCR-RFLP analyses also showed a high diversity among field isolates of M. hyopneumoniae with a greater homogeneity within the same herd. Almost half of the field isolates presented less than 55% homology with selected vaccine and reference strains. PMID:24345410

  19. Transposon mutagenesis in Mycoplasma hyopneumoniae using a novel mariner-based system for generating random mutations

    PubMed Central

    2013-01-01

    Mycoplasma hyopneumoniae is the cause of enzootic pneumonia in pigs, a chronic respiratory disease associated with significant economic losses to swine producers worldwide. The molecular pathogenesis of infection is poorly understood due to the lack of genetic tools to allow manipulation of the organism and more generally for the Mycoplasma genus. The objective of this study was to develop a system for generating random transposon insertion mutants in M. hyopneumoniae that could prove a powerful tool in enabling the pathogenesis of infection to be unraveled. A novel delivery vector was constructed containing a hyperactive C9 mutant of the Himar1 transposase along with a mini transposon containing the tetracycline resistance cassette, tetM. M. hyopneumoniae strain 232 was electroporated with the construct and tetM-expressing transformants selected on agar containing tetracycline. Individual transformants contained single transposon insertions that were stable upon serial passages in broth medium. The insertion sites of 44 individual transformants were determined and confirmed disruption of several M. hyopneumoniae genes. A large pool of over 10 000 mutants was generated that should allow saturation of the M. hyopneumoniae strain 232 genome. This is the first time that transposon mutagenesis has been demonstrated in this important pathogen and could be generally applied for other Mycoplasma species that are intractable to genetic manipulation. The ability to generate random mutant libraries is a powerful tool in the further study of the pathogenesis of this important swine pathogen. PMID:24359443

  20. Mycoplasma pneumoniae encephalitis in childhood.

    PubMed

    Lin, Wen-Chuan; Lee, Ping-Ing; Lu, Chun-Yi; Hsieh, Yu-Chia; Lai, Hsin-Pao; Lee, Chin-Yun; Huang, Li-Min

    2002-09-01

    Mycoplasma pneumoniae is an important etiologic agent of acute childhood encephalitis. We retrospectively reviewed 17 cases of M. pneumoniae encephalitis at the Pediatric Department of the National Taiwan University Hospital from April 1997 through March 2000. These cases were diagnosed as having positive immunoglobulin M antibodies (94%), a minimum 4-fold change of complement-fixation antibody titers (47%), or nested polymerase chain reaction. The ages of these patients ranged from 1.5 to 10.9 years (mean, 5.3 years) with a male-to-female ratio of 8:9. The clinical manifestations included fever (94%), altered consciousness (65%), seizure (41%), personality or behavior changes (29%), meningeal sign (24%), visual hallucination (24%), ataxia (12%), Guillain-Barré syndrome (6%), blurred vision (6%), and aphasia (6%). Respiratory symptoms and signs were found in 76% of the patients. Abnormal electroencephalogram and neuroimage were observed in all cases, while abnormal cerebrospinal fluid examination was noted in about one-third of the patients. Five (29%) patients required intensive care because of intractable seizure or respiratory failure. Fourteen (82%) patients recovered completely, but 3 (18%) had sequelae including epilepsy, hydrocephalus, and global neurologic deficits with brain stem dysfunction. In Taiwan, M. pneumoniae should be considered an etiologic pathogen of acute childhood encephalitis if fever and respiratory symptoms and signs are observed with or without abnormal cerebrospinal fluid findings. Supportive treatment is the basis of management. PMID:12380790

  1. Immunological Analysis of Mycoplasma Membranes

    PubMed Central

    Kahane, Itzhak; Razin, Shmuel

    1969-01-01

    The antigens responsible for the production of antibodies to Mycoplasma laidlawii and M. gallisepticum causing growth and metabolic inhibition of these organisms were localized in the cell membrane. Various membrane fractions were tested for serological activity. Membrane lipids were completely or almost completely inactive, whereas several preparations of defatted membrane proteins retained some serological activity, shown by their ability to stimulate metabolic inhibition antibody in rabbits and to adsorb metabolic inhibition antibody and form precipitation lines with an antiserum to the membrane. When the membranes were heated to 65 C for 1 hr, they virtually lost their ability to adsorb metabolic inhibition antibody, which suggests that the antigenic determinants are proteins. Serological activity was retained in reaggregated membranes obtained by dialysis against Mg2+ of membranes solubilized in sodium dodecyl sulfate. The amount of solubilized membrane protein and lipid incorporated into the reaggregated membranes could be regulated by varying the Mg2+ concentration. As the serological tests indicated that the various membrane antigens were selectively incorporated into the different reaggregated membranes, the use of controlled reaggregation of solubilized membranes is suggested as a new tool for the fractionation and antigenic analysis of membrane proteins. Images PMID:4981055

  2. Observations on the occurrence of mycoplasmas in the central nervous system of some laboratory animals.

    PubMed

    Taylor-Robinson, D; Furr, P M

    1981-07-01

    Mycoplasma pulmonis was isolated from the brains of 6 (23%) of 26 mice which had a naturally-occurring respiratory infection with this mycoplasma, and from the brains of 6 (8%) of 71 mice which had been inoculated intranasally or intravenously. The incidence of natural infection was greater in older mice, but there was no obvious mouse strain difference except for higher incidence in athymic nudes. There was no evidence that the organisms passed the blood-brain barrier. Some isolations, especially from nudes, may have been extraneous contaminants, as these were fewer when the mouse skulls were sterilized with ignited methanol. M. pneumoniae was not isolated from the brains of 14 hamsters which had a respiratory infection after intranasal inoculation nor were ureaplasmas isolated from the cerebrospinal fluids of 12 marmosets with a natural oropharyngeal infection. The aetiology of M. pneumoniae encephalitis in man is discussed. PMID:6793791

  3. Hydrogen Peroxide Production from Glycerol Metabolism Is Dispensable for Virulence of Mycoplasma gallisepticum in the Tracheas of Chickens

    PubMed Central

    Szczepanek, S. M.; Boccaccio, M.; Pflaum, K.; Liao, X.

    2014-01-01

    Hydrogen peroxide (H2O2) is a by-product of glycerol metabolism in mycoplasmas and has been shown to cause cytotoxicity for cocultured eukaryotic cells. There appears to be selective pressure for mycoplasmas to retain the genes needed for glycerol metabolism. This has generated interest and speculation as to their function during infection. However, the actual effects of glycerol metabolism and H2O2 production on virulence in vivo have never been assessed in any Mycoplasma species. To this end, we determined that the wild-type (WT) Rlow strain of the avian pathogen Mycoplasma gallisepticum is capable of producing H2O2 when grown in glycerol and is cytotoxic to eukaryotic cells in culture. Transposon mutants with mutations in the genes present in the glycerol transport and utilization pathway, namely, glpO, glpK, and glpF, were identified. All mutants assessed were incapable of producing H2O2 and were not cytotoxic when grown in glycerol. We also determined that vaccine strains ts-11 and 6/85 produce little to no H2O2 when grown in glycerol, while the naturally attenuated F strain does produce H2O2. Chickens were infected with one of two glpO mutants, a glpK mutant, Rlow, or growth medium, and tracheal mucosal thickness and lesion scores were assessed. Interestingly, all glp mutants were reproducibly virulent in the respiratory tracts of the chickens. Thus, there appears to be no link between glycerol metabolism/H2O2 production/cytotoxicity and virulence for this Mycoplasma species in its natural host. However, it is possible that glycerol metabolism is required by M. gallisepticum in a niche that we have yet to study. PMID:25156740

  4. Rescue of mutant fitness defects using in vitro reconstituted designer transposons in Mycoplasma mycoides

    PubMed Central

    Karas, Bogumil J.; Wise, Kim S.; Sun, Lijie; Venter, J. Craig; Glass, John I.; Hutchison, Clyde A.; Smith, Hamilton O.; Suzuki, Yo

    2014-01-01

    With only hundreds of genes contained within their genomes, mycoplasmas have become model organisms for precise understanding of cellular processes, as well as platform organisms for predictable engineering of microbial functions for mission-critical applications. Despite the availability of “whole genome writing” in Mycoplasma mycoides, some traditional methods for genetic engineering are underdeveloped in mycoplasmas. Here we demonstrate two facile transposon-mediated approaches for introducing genes into the synthetic cell based on M. mycoides. The marker-less approach involves preparing a fragment containing only a small genomic region of interest with flanking transposase-binding sites, followed by in vitro transposase loading and introduction into the cells. The marker-driven approach involves cloning an open reading frame (ORF) of interest into a vector containing a marker for mycoplasma transformation, as well as sites for transposase loading and random genomic integration. An innovative feature of this construct is to use a single promoter to express the transformation marker and the introduced ORF. The marker-driven approach can be conveniently applied to any exogenous or synthetic gene without any information on the effect of the gene on the strain, whereas the marker-less approach requires that the fragment has a recognizable effect. Using the marker-less method, we found that a region containing the nusG gene rescues a slow growth phenotype of a strain containing a larger deletion encompassing this gene. Using the marker-driven approach, we better defined this finding, thereby establishing that nusG is required for a normal growth rate in synthetic M. mycoides. These methods are suitable for complementation tests to identify genes responsible for assorted functions lacking in deletion mutants. These approaches are also expected to facilitate rapid testing of various natural and engineered genes or gene clusters from numerous sources in M. mycoides. PMID:25101070

  5. Plasminogen Binding and Activation by Mycoplasma fermentans

    Microsoft Academic Search

    AMICHAI YAVLOVICH; ABD A.-R. HIGAZI; SHLOMO ROTTEM

    2001-01-01

    The binding of plasminogen to Mycoplasma fermentans was studied by an immunoblot analysis and by a binding assay using iodine-labeled plasminogen. The binding of 125 I-labeled plasminogen was inhibited by unlabeled plasminogen, lysine, and lysine analog «-aminocaproic acid. Partial inhibition was obtained by a plasminogen fragment containing kringles 1 to 3 whereas almost no inhibition was observed with a fragment

  6. Effect of exogenous adminstration of antioxidants on Blastocystis hominis in vivo.

    PubMed

    Helmy, Moshera M; Hussein, Eman M; El-Moamly, Amal A; Eida, Amany M; Eida, Omima M; Salem, Attia M

    2008-04-01

    The effect of exogenous administration of antioxidant (Anttox) on the course of B. hominis in experimentally infected mice was studied. B. hominis isolates were obtained from 10 gastrointestinal symptomatic adult patients. Three groups of 30 infected mice (3/isolate) were used. GI was untreated infected, GII was treated by antox for 4 weeks after infection diagnosis (treatment strategy), and GIII antox treated by for 4 weeks before infection (prophylactic strategy). Mild pathological changes were detected on 13.4%, 19.9% & 86.8% of mice in Gs I, II & III, respectively. Moderate pathological changes were found in 29.9%, 26.6% & 6.6% of mice in Gs I, II & III, respectively. While, the majority of severe pathological changes were in Gs I & II (56.7% & 53.5%) as compared to GIII (6.6%). Meanwhile, 86.8% of mice in GIII had B. hominis forms > 10/high power field compared to 3.3% in Gs I & II, respectively. Although 19.8% of mice in GII were positive for B. hominis by direct smear, no growth resulted in vitro and all the forms were non-viable by using neutral red stain. All the differences were statistically significant. So, antioxidant exacerbated B. hominis intensity but it decreased the pathological changes. PMID:19143124

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

    PubMed

    Gharaibeh, Saad; Al-Rashdan, Mohammad

    2011-06-01

    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 ?g/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. PMID:21382675

  8. In vitro sensitivity of Blastocystis hominis to garlic, ginger, white cumin, and black pepper used in diet.

    PubMed

    Yakoob, Javed; Abbas, Zaigham; Beg, Muhammad Asim; Naz, Shagufta; Awan, Safia; Hamid, Saeed; Jafri, Wasim

    2011-08-01

    To determine the growth pattern and in vitro susceptibility of Blastocystis hominis to metronidazole (MTZ), garlic, ginger, white cumin, and black pepper. Stool specimens were collected from 16 irritable bowel syndrome (IBS) and 10 controls between July-November 2010. Stool microscopy and culture for B. hominis was performed. Drug susceptibility assays was done using 0.01 and 0.1 mg/ml of MTZ, garlic, ginger, white cumin, and black pepper. Effect was assessed on B. hominis culture after 48 h. Stool DNA was extracted using stool DNA extraction kit (Qiagen) and polymerase chain reaction (PCR) done using subtype-specific sequence-tagged-site primers. B. hominis genotype 3 and coinfection of 1 and 3 tended to grow well in culture compared to isolated type 1 infection. Exposed to MTZ at a concentration of 0.01 mg/ml, 38% (6/16) B. hominis from IBS did not grow in culture compared to 100% (10/10) of B. hominis from control (p = 0.001). When they were exposed to MTZ at 0.1 mg/ml, 56% (9/16) B. hominis from IBS did not grow in cultures compared to 100% (10/10) from control (p = 0.01). Forty-four percent (7/16) B. hominis from IBS did not grow in culture compared to 100% (10/10) B. hominis from control when exposed to garlic at a concentration of 0.01 mg/ml (p = 0.003) and following exposure to garlic at 0.1 mg/ml, 38% (6/16) B. hominis from IBS did not grow in cultures compared to 100% (10/10) from control (p = 0.001). B. hominis isolates from IBS had a cell count of 6,625 at a MTZ concentration of 0.01 mg/ml that reduced to 1,250 as MTZ concentration was increased to 0.1 mg/ml (p = 0.08). B. hominis from IBS with a mean cell count of 3 × 10(5) at baseline decreased to 1 × 10(4) when exposed to garlic at 0.01 mg/ml (p < 0.001) and to 1 × 10(3) (p < 0.001) when garlic was 0.1 mg/ml. B. hominis from IBS cell count decreased to 1 × 10(5) when exposed to white cumin at 0.01 mg/ml (p = 0.01) and to 1 × 10(5) (p < 0.001) when white cumin was 0.1 mg/ml. Exposed to black pepper at 0.1 mg/ml, cell count of B. hominis from IBS decreased to 1 × 10(5) (p = 0.01). B. hominis from IBS decreased to 1.3 × 10(5) exposed to ginger at 0.01 mg/ml (p = 0.001). B. hominis isolates were mostly genotypes 3, type 1 and 3 coinfection, and non-typeable B. hominis isolates. B. hominis isolates from IBS mostly genotype 1 demonstrated an increased sensitivity to garlic at 0.01 mg/ml with a B. hominis cell count of 3,714 compared to 6,142 when exposed to 0.01 mg/ml of MTZ. However, this sensitivity did not increase as garlic concentration was increased to 0.1 mg/ml, for B. hominis cell count was 6,000 compared to 1,428 as MTZ was increased to 0.1 mg/ml. PMID:21431384

  9. Prevalence of mycoplasmas in eggs from birds of prey using culture and a genus-specific mycoplasma polymerase chain reaction.

    PubMed

    Lierz, M; Hagen, N; Harcourt-Brown, N; Hernandez-Divers, S J; Lüschow, D; Hafez, H M

    2007-04-01

    Mycoplasmas are commensals and pathogens of different avian species, especially poultry and passeriforms. The role of mycoplasmas in raptors has not yet been completely determined, and especially not the possibility of vertical transmission. Therefore 424 raptor eggs were examined for the occurrence of mycoplasmas using culture, and 155 of these eggs with a Mycoplasma genus-specific polymerase chain reaction (PCR) assay. This PCR was tested for its sensitivity and specificity, especially for use in a bird population of unknown mycoplasma status (prevalence and species). The size of the amplified PCR product was large (1013 base pairs) to enable use of the product for species differentiation by sequencing. Culture and PCR yielded only one positive result, in an egg of a Northern Goshawk (Accipiter gentilis). The isolate was identified as Mycoplasma lipofaciens using an immunobinding assay, as well as by sequencing part of its 16S rRNA gene. PMID:17479375

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

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375 Mycoplasma spp. serological reagents. (a) Identification....

  11. Effects of carbohydrates from citrus pulp and hominy feed on microbial fermentation in continuous culture.

    PubMed

    Ariza, P; Bach, A; Stern, M D; Hall, M B

    2001-10-01

    Eight dual-flow continuous-culture fermenters were used to evaluate the effect of neutral detergent-soluble carbohydrates (NDSC) on fermentation by ruminal microorganisms. Citrus pulp and hominy feed were added to a basal diet as sources of NDSC, with citrus pulp providing neutral detergent-soluble fiber (NDSF) in the form of pectic substances and with hominy feed in the form of starch. The basal diet contained 26.7% corn silage, 6.0% alfalfa hay and 3.8% cottonseed hulls on a DM basis. The dried citrus pulp diet contained on a DM basis 17.2% CP, 34.7% NDF, 33.7% NDSC, and 14.4% NDSF, whereas the hominy feed diet contained 17.9% CP, 33.2% NDF, 35.9% NDSC, and 8.8% NDSF. Organic matter, DM, and NDF and ADF digestion were not affected by source of carbohydrate. Ammonia N concentration was greater (P < 0.05) for the hominy feed diet (14.2 mg/100 mL) than for the dried citrus pulp diet (9.3 mg/100 mL). Total N, nonammonia N, microbial N, and dietary N flows were not affected by treatments; however, the efficiency of microbial protein synthesis was greater (P = 0.055) for the dried citrus pulp diet than for the hominy feed diet (30.6 vs 27.8 g of bacterial N/kg of OM truly digested). Results from this experiment indicate that NDSF from citrus pulp can provide similar sources of energy compared with starch from hominy feed to support ruminal microbial growth. PMID:11721852

  12. Comparative Geno-Plasticity Analysis of Mycoplasma bovis HB0801 (Chinese Isolate)

    PubMed Central

    Qi, Jingjing; Guo, Aizhen; Cui, Peng; Chen, Yingyu; Mustafa, Riaz; Ba, Xiaoliang; Hu, Changmin; Bai, Zhidi; Chen, Xi; Shi, Lei; Chen, Huanchun

    2012-01-01

    Mycoplasma bovis pneumonia in cattle has been epidemic in China since 2008. To investigate M. bovis pathogenesis, we completed genome sequencing of strain HB0801 isolated from a lesioned bovine lung from Hubei, China. The genomic plasticity was determined by comparing HB0801 with M. bovis strain ATCC® 25523™/PG45 from cow mastitis milk, Chinese strain Hubei-1 from lesioned lung tissue, and 16 other Mycoplasmas species. Compared to PG45, the genome size of HB0801 was reduced by 11.7 kb. Furthermore, a large chromosome inversion (580 kb) was confirmed in all Chinese isolates including HB0801, HB1007, a strain from cow mastitis milk, and Hubei-1. In addition, the variable surface lipoproteins (vsp) gene cluster existed in HB0801, but contained less than half of the genes, and had poor identity to that in PG45, but they had conserved structures. Further inter-strain comparisons revealed other mechanisms of gene acquisition and loss in HB0801 that primarily involved insertion sequence (IS) elements, integrative conjugative element, restriction and modification systems, and some lipoproteins and transmembrane proteins. Subsequently, PG45 and HB0801 virulence in cattle was compared. Results indicated that both strains were pathogenic to cattle. The scores of gross pathological assessment for the control group, and the PG45- and HB0801-infected groups were 3, 13 and 9, respectively. Meanwhile the scores of lung lesion for these three groups were 36, 70, and 69, respectively. In addition, immunohistochemistry detection demonstrated that both strains were similarly distributed in lungs and lymph nodes. Although PG45 showed slightly higher virulence in calves than HB0801, there was no statistical difference between the strains (P>0.05). Compared to Hubei-1, a total of 122 SNP loci were disclosed in HB0801. In conclusion, although genomic plasticity was thought to be an evolutionary advantage, it did not apparently affect virulence of M. bovis strains in cattle. PMID:22693604

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

    Microsoft Academic Search

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

    2007-01-01

    The in vitro effect of six Jordanian traditional medicine plant methanolic extracts were tested against 32 isolates of Mycoplasma species; Mycoplasma mycoides subsp. mycoides LC (6), Mycoplasma capricolum subsp. capricolum (8) and M. putrefaciens (18), all isolated from either nasal swabs or milk, from sheep and goats in different regions in Jordan. All Mycoplasma species showed susceptibility to Artemisia herba-alba

  14. Induction of antitumor activity in macrophages by mycoplasmas in concert with interferon

    Microsoft Academic Search

    Kazuko Uno; Morio Takema; Shigetaka Hidaka; Reishi Tanaka; Takao Konishi; Takuma Kato; Shinji Nakamura; Shigeru Muramatsu

    1990-01-01

    Summary The in vitro growth of tumor cells infected with mycoplasmas was suppressed by macrophages pretreated with interferon (IFN), but the growth of mycoplasma-free tumor cells was not suppressed. Pretreatment of macrophages with IFN plus mycoplasmas or their soluble factors either simultaneously or sequentially, IFN first and mycoplasmas second, but not in the reverse order, was effective in activating macrophages

  15. Characterization of western X-disease mycoplasma-like organisms

    SciTech Connect

    Kirkpatrick, B.C.

    1986-01-01

    The causal agent of western X-disease, an important disease of cherry (Prunus avium) and peach (Prunus persica) in the western United States, was shown to be a non-culturable, mycoplasma-like organism (WX-MLO). Procedures were developed to purify WX-MLOs from celery and leafhoppers infected with a greenhouse-maintained isolate of the peach yellow leaf roll (ghPYLR) strain of western X-disease. WX-MLOs, purified from ghPYLR-infected leafhoppers, elicited the production of specific antisera (WX antisera) when injected into rabbits. When used in an enzyme-linked immunosorbent assay (ELISA), WX antisera quantitatively detected WX-MLOs in celery, periwinkle, and leafhoppers experimentally infected with either ghPYLR or the Green Valley (GVX) strain of western X-disease. Recombinant clones were screened by colony, dot and southern hybridizations using /sup 32/P-nick translated DNA extracted from healthy and ghPYLR-infected celery and leafhoppers. Twenty-four clones were identified which hybridized with DNA from diseased but not healthy hosts. DNA hybridization assays, using radiolabeled, cloned WX-MLO DNA, readily detected WX-MLOs in celery, periwinkle, and leafhoppers infected with either GVX or ghPYLR and in cherry and peach with symptoms of GVX.

  16. The Minimal Gene Complement of Mycoplasma genitalium

    Microsoft Academic Search

    Claire M. Fraser; Jeannine D. Gocayne; Owen White; Mark D. Adams; Rebecca A. Clayton; Robert D. Fleischmann; Carol J. Bult; Anthony R. Kerlavage; Granger Sutton; Jenny M. Kelley; Janice L. Fritchman; Janice F. Weidman; Keith V. Small; Mina Sandusky; Joyce Furhmann; David Nguyen; Teresa R. Utterback; Deborah M. Saudek; Cheryl A. Phillips; Joseph M. Merrick; Jean-Francois Tomb; Brian A. Dougherty; Kenneth F. Bott; Ping-Chuan Hu; Thomas S. Lucier; Scott N. Peterson; Hamilton O. Smith; Clyde A. Hutchison III; J. Craig Venter

    1995-01-01

    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

  17. Identification and Subtyping of Clinically Relevant Human and Ruminant Mycoplasmas by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

    PubMed Central

    Renaudin, H.; Cauvin, E.; Del Prá Netto Machado, L.; Tricot, A.; Benoit, F.; Treilles, M.; Bébéar, C.

    2013-01-01

    Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) recently emerged as a technology for the identification of bacteria. In this study, we aimed to evaluate its applicability to human and ruminant mycoplasmal identification, which can be demanding and time-consuming when using phenotypic or molecular methods. In addition, MALDI-TOF MS was tested as a subtyping tool for certain species. A total of 29 main spectra (MSP) from 10 human and 13 ruminant mycoplasma (sub)species were included in a mycoplasma MSP database to complete the Bruker MALDI Biotyper database. After broth culture and protein extraction, MALDI-TOF MS was applied for the identification of 119 human and 143 ruminant clinical isolates that were previously identified by antigenic or molecular methods and for subcultures of 73 ruminant clinical specimens that potentially contained several mycoplasma species. MALDI-TOF MS resulted in accurate (sub)species-level identification with a score of ?1.700 for 96% (251/262) of the isolates. The phylogenetically closest (sub)species were unequivocally distinguished. Although mixtures of the strains were reliably detected up to a certain cellular ratio, only the predominant species was identified from the cultures of polymicrobial clinical specimens. For typing purposes, MALDI-TOF MS proved to cluster Mycoplasma bovis and Mycoplasma agalactiae isolates by their year of isolation and genome profiles, respectively, and Mycoplasma pneumoniae isolates by their adhesin P1 type. In conclusion, MALDI-TOF MS is a rapid, reliable, and cost-effective method for the routine identification of high-density growing mycoplasmal species and shows promising prospects for its capacity for strain typing. PMID:23903545

  18. Identification and subtyping of clinically relevant human and ruminant mycoplasmas by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry.

    PubMed

    Pereyre, S; Tardy, F; Renaudin, H; Cauvin, E; Del Prá Netto Machado, L; Tricot, A; Benoit, F; Treilles, M; Bébéar, C

    2013-10-01

    Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) recently emerged as a technology for the identification of bacteria. In this study, we aimed to evaluate its applicability to human and ruminant mycoplasmal identification, which can be demanding and time-consuming when using phenotypic or molecular methods. In addition, MALDI-TOF MS was tested as a subtyping tool for certain species. A total of 29 main spectra (MSP) from 10 human and 13 ruminant mycoplasma (sub)species were included in a mycoplasma MSP database to complete the Bruker MALDI Biotyper database. After broth culture and protein extraction, MALDI-TOF MS was applied for the identification of 119 human and 143 ruminant clinical isolates that were previously identified by antigenic or molecular methods and for subcultures of 73 ruminant clinical specimens that potentially contained several mycoplasma species. MALDI-TOF MS resulted in accurate (sub)species-level identification with a score of ?1.700 for 96% (251/262) of the isolates. The phylogenetically closest (sub)species were unequivocally distinguished. Although mixtures of the strains were reliably detected up to a certain cellular ratio, only the predominant species was identified from the cultures of polymicrobial clinical specimens. For typing purposes, MALDI-TOF MS proved to cluster Mycoplasma bovis and Mycoplasma agalactiae isolates by their year of isolation and genome profiles, respectively, and Mycoplasma pneumoniae isolates by their adhesin P1 type. In conclusion, MALDI-TOF MS is a rapid, reliable, and cost-effective method for the routine identification of high-density growing mycoplasmal species and shows promising prospects for its capacity for strain typing. PMID:23903545

  19. Mycoplasma ovis in captive cervids: Prevalence, molecular characterization and phylogeny

    Microsoft Academic Search

    Ana Laura Grazziotin; Andrea Pires Santos; Ana Marcia Sa Guimaraes; Ahmed Mohamed; Zalmir Silvino Cubas; Marcos Jose de Oliveira; Leonilda Correia dos Santos; Wanderlei de Moraes; Rafael Felipe da Costa Vieira; Lucelia Donatti; Ivan Roque de Barros Filho; Alexander Welker Biondo; Joanne Belle Messick

    2011-01-01

    Hemotrophic mycoplasmas (hemoplasmas) are bacteria that attach to red blood cells of mammals, leading to acute and\\/or subclinical disease in infected animals. It has been suggested that Mycoplasma ovis, a hemoplasma that infects sheep and goats worldwide, may also infect deer. The aim of this study was to evaluate whether South American deer are infected with M. ovis. EDTA-anticoagulated blood

  20. Mycoplasma pneumonia infection and asthma: A clinical study

    PubMed Central

    Gao, Shuncui; Wang, Lingcheng; Zhu, Wei; Jiang, Jie

    2015-01-01

    Objectives: To evaluate the correlation between mycoplasma pneumonia infection and the severity of asthma as well as asthma control, to help physicians in respiratory department better make treatment strategies. Methods: Since January 2012 to May 2014, we consecutively recruited 149 out-patients diagnosed with asthma in acute or convalescent phase from the department of respiratory medicine of our hospital. The pulmonary function tests, sputum induction examination, measurement of IgM, IgG and IgE in serum, evaluation of asthma control were carried out for all the included patients. Results: In 78 cases with asthma in acute phase, mycoplasma pneumonia infection was confirmed in 38 cases (48.71%), and in 71 cases in stable state, mycoplasma pneumonia infection was confirmed in 22 cases (30.98%). There was significant difference in the rate of mycoplasma pneumonia infection between the two groups (p<0.05). The FEV1% Pred and ACT scores were significantly lower in mycoplasma pneumonia infection cases than those in no mycoplasma pneumonia infection cases (p<0.05), while the eosinophil count and IgE in serum were significantly higher in mycoplasma pneumonia infection cases (p<0.05). Conclusions: Mycoplasma pneumonia infection may play more important role in the occurrence of acute asthma, and it can lead to decreased pulmonary function, difficulty in controlling asthma and more severe airway inflammation.

  1. TRITRICHOMONAS FOETUS AND NOT PENTATRICHOMONAS HOMINIS IS THE ETIOLOGIC AGENT OF FELINE TRICHOMONAL DIARRHEA

    Microsoft Academic Search

    Michael G. Levy; Jody L. Gookin; Matthew Poore; Adam J. Birkenheuer; Michael J. Dykstra; R. Wayne Litaker

    2003-01-01

    Recently, several investigators have reported large-bowel diarrhea in cats associated with intestinal trichomonad parasites. These reports have presumptively identified the flagellates as Pentatrichomonas hominis , an organism putatively capable of infecting the intestinal tracts of a number of mammalian hosts, including cats, dogs, and man. The purpose of the present study was to determine the identity of this recently recognized

  2. Direct comparison of selected methods for genetic categorisation of Cryptosporidium parvum and Cryptosporidium hominis species

    Microsoft Academic Search

    Rachel M. Chalmers; Christobel Ferguson; Simone Cacciò; Robin B. Gasser; Youssef G. Abs EL-Osta; Leo Heijnen; Lihua Xiao; Kristin Elwin; Stephen Hadfield; Martha Sinclair; Melita Stevens

    2005-01-01

    A study was undertaken to compare the performance of five different molecular methods (available in four different laboratories) for the identification of Cryptosporidium parvum and Cryptosporidium hominis and the detection of genetic variation within each of these species. The same panel of oocyst DNA samples derived from faeces (n=54; coded blindly) was sent for analysis by: (i) DNA sequence analysis

  3. Molecular Epidemiology of Cases of Mycoplasma californicum Infection in Japan

    PubMed Central

    Suzuki, Kan-ichiro; Hanyu, Hideki; Itoh, Megumi; Higuchi, Hidetoshi; Kobayashi, Hideki

    2014-01-01

    Bovine mastitis due to Mycoplasma californicum is often accompanied by huge economic losses, and the disease spreads very quickly. An appropriate molecular epidemiological analysis is needed to prevent and control infectious disease, but molecular epidemiological analysis methods for M. californicum have not yet been reported. Here we developed a combination of multiple-locus variable-number tandem repeat analysis (MLVA) and pulsed-field gel electrophoresis (PFGE) methods, which are common genotyping methods for various bacteria, for M. californicum. The MLVA is based on four interspersed repeat units that were found in the M. californicum genome data. The MLVA using these repeat units showed sufficient discriminatory power for a molecular epidemiological analysis; i.e., a Hunter-Gaston diversity index (HGDI) of 0.949, against M. californicum strains in Japan and M. californicum strain ATCC 33461. The PFGE for M. californicum also showed sufficient discriminatory power, with an HGDI of 0.985. Strain ATCC 33461 showed MLVA profiles and pulsotypes that differed greatly from those of strains from Japan. These results indicate that MLVA and PFGE are good tools for identifying M. californicum transmission events more accurately. Our combined MLVA and PFGE analysis suggests the persistence of M. californicum infection among herds in a specific area for a long period of time, as well as the movement of cows and heifers accompanying the expansion of M. californicum infection. Failure to identify asymptomatic infected cows is suspected as one of the central causes of the present M. californicum infection scenario in Japan. PMID:25281385

  4. Mycoplasma removal: simple curative methods for viral supernatants.

    PubMed

    Baronti, C; Pastorino, B; Charrel, R; de Lamballerie, X

    2013-02-01

    As a partner of the European Virus Archive (EVA) FP7 infrastructure, our research group is maintaining and developing a large virus collection. To meet the standards of the quality management system adopted by all European Virus Archive partners, the detection and eradication of mycoplasma in cell culture supernatants (stored at -80°C or freeze-dried) has to be improved. Although the methods for mycoplasma elimination from infected cell lines were largely described, the decontamination procedures of precious cell culture supernatants was poorly documented. In this study, a large panel of mycoplasma-contaminated virus stocks (enveloped and non enveloped, RNA and DNA viruses) was tested successfully for mycoplasma removal using two simple optimized methods. These easy-to-perform protocols, using respectively Plasmocin™ (InvivoGen, Cayla, France) and chloroform, were shown to remove mycoplasma completely from cell supernatant without incidence in viral infectivity. PMID:23117065

  5. Viability of Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri in goat milk samples stored under different conditions

    Microsoft Academic Search

    Joaquín Amores; Antonio Sánchez; Ángel Gómez Martín; Juan C. Corrales; Antonio Contreras; Christian de la Fe

    2010-01-01

    Control programs for contagious agalactia (CA) involve monitoring milk samples to detect this disease. This study was designed to establish the effects of the preservatives generally used in dairy laboratories and storage temperature on the viability of Mycoplasma (M.) agalactiae (Ma) and M. mycoides subsp. capri (Mmc) in goat milk samples. In total, 1440 determinations were conducted for each mycoplasma

  6. Interlaboratory comparison of ability to detect nucleic acid of Mycoplasma gallisepticum and Mycoplasma synoviae by polymerase chain reaction

    Microsoft Academic Search

    M. Hess; C. Neubauer; R. Hackl

    2007-01-01

    In recent years polymerase chain reaction (PCR) assays have become widely used as methods to confirm the presence of Mycoplasma gallisepticum and Mycoplasma synoviae in poultry flocks, but there has been limited standardization of the protocols used. Thirteen laboratories from five different countries participated in an interlaboratory comparison of detection of M. gallisepticum and M. synoviae DNA by PCR in

  7. Human Coinfection with Bartonella henselae and Two Hemotropic Mycoplasma Variants Resembling Mycoplasma ovis?

    PubMed Central

    Sykes, Jane E.; Lindsay, LeAnn L.; Maggi, Ricardo G.; Breitschwerdt, Edward B.

    2010-01-01

    Two variants of an organism resembling the ovine hemoplasma, Mycoplasma ovis, were detected by PCR in blood samples from a veterinarian in Texas. Coinfection with similar variants has been described in sheep. This represents the first report of human infection with this organism. The veterinarian was coinfected with Bartonella henselae. PMID:20702675

  8. Relationship between Mutations in Dihydropteroate Synthase of Pneumocystis carinii f. sp. hominis Isolates in Japan and Resistance to Sulfonamide Therapy

    PubMed Central

    Takahashi, Takashi; Hosoya, Noriaki; Endo, Tokiomi; Nakamura, Tetsuya; Sakashita, Hiroyuki; Kimura, Kyoko; Ohnishi, Kenji; Nakamura, Yoshikazu; Iwamoto, Aikichi

    2000-01-01

    We examined mutations in the dihydropteroate synthase (DHPS) genes of Pneumocystis carinii f. sp. hominis (P. carinii) strains isolated from 24 patients with P. carinii pneumonia (PCP) in Japan. DHPS mutations were identified at amino acid positions 55 and/or 57 in isolates from 6 (25.0%) of 24 patients. The underlying diseases for these six patients were human immunodeficiency virus type 1 infection (n = 4) or malignant lymphoma (n = 2). This frequency was almost the same as those reported in Denmark and the United States. None of the six patients whose isolates had DHPS mutations were recently exposed to sulfa drugs before they developed the current episode of PCP, suggesting that DHPS mutations not only are selected by the pressure of sulfa agents but may be incidentally acquired. Co-trimoxazole treatment failed more frequently in patients whose isolates had DHPS mutations than in those whose isolates had wild-type DHPS (n = 4 [100%] versus n = 2 [11.1%]; P = 0.002). Our results thus suggest that DHPS mutations may contribute to failures of co-trimoxazole treatment for PCP. PMID:10970350

  9. Synthesis, integration, and restriction and modification of mycoplasma virus L2 DNA

    SciTech Connect

    Dybvig, K.

    1981-01-01

    Mycoplasma virus L2 is an enveloped, nonlytic virus containing double-stranded, superhelical DNA. The L2 virion contains about 7 to 8 major proteins identified by SDS-polyacrylamide gel electrophoresis, but the virion has no discernible capsid structure. It has been suggested that the L2 virion is a DNA-protein condensation surrounded by a lipid-protein membrane. The host for mycoplasma virus L2 is Acholeplasma laidlawii. A. laidlawii has no cell wall and contains a small genome, 1 x 10/sup 9/ daltons, which is two to three times smaller than that of most bacteria. Infection of A. laidlawii by L2 is nonlytic. The studies in this thesis show that L2 DNA synthesis begins at about 1 hour of infection and lasts throughout the infection. Viral DNA synthesis is inhibited by chloramphenicol, streptomycin, and novobiocin. Packaging of L2 DNA into progeny virus is also inhibited by chloramphenicol and novobiocin. It is concluded that protein synthesis and probably DNA gyrase activity are required for L2 DNA synthesis, and for packaging of L2 DNA into progeny virus. DNA-DNA hybridization studies demonstrate that L2 DNA integrates into the host cell during infection, and subsequent to infection the cells are mycoplasma virus L2 lysogens. The viral site of integration has been roughly mapped. L2 virus is restricted and modified by A. laidlawii strains JA1 and K2. The nature of the modification in strain K2 has been elucidated. Two L2 variants containing insertions in the viral DNA were identified in these studies. Restriction endonuclease cleavage maps of these variants have been determined. DNA from L2 and another isolate of L2, MV-Lg-L 172, are compared in these studies. 74 references, 33 figures, 6 tables. (ACR)

  10. Clinical efficacy of Saccharomyces boulardii or metronidazole in symptomatic children with Blastocystis hominis infection.

    PubMed

    Dinleyici, Ener Cagri; Eren, Makbule; Dogan, Nihal; Reyhanioglu, Serap; Yargic, Zeynel Abidin; Vandenplas, Yvan

    2011-03-01

    Although many Blastocystis infections remain asymptomatic, recent data suggest it also causes frequent symptoms. Therapy should be limited to patients with persistent symptoms and a complete workup for alternative etiologies. The goal of this study was to compare the natural evolution (no treatment) to the efficacy of Saccharomyces boulardii (S. boulardii) or metronidazole for the duration of diarrhea and the duration of colonization in children with gastrointestinal symptoms and positive stool examination for Blastocystis hominis. This randomized single-blinded clinical trial included children presenting with gastrointestinal symptoms (abdominal pain, diarrhea, nausea-vomiting, flatulence) more than 2 weeks and confirmed B. hominis by stool examination (B. hominis cysts in the stool with microscopic examination of the fresh stool). The primary end points were clinical evaluation and result of microscopic stool examination at day 15. Secondary end points were the same end points at day 30. Randomization was performed by alternating inclusion: group A, S. boulardii (250 mg twice a day, Reflor®) during 10 days; group B, metronidazole (30 mg/kg twice daily) for 10 days; group C, no treatment. At day 15 and 30 after inclusion, the patients were re-evaluated, and stool samples were examined microscopically. On day 15, children that were still symptomatic and/or were still B. hominis-infected in group C were treated with metronidazole for 10 days. There was no statistically significant difference between the three study groups for age, gender, and the presence of diarrhea and abdominal pain. On day 15, clinical cure was observed in 77.7% in group A (n, 18); in 66.6% in group B (n, 15); and 40% in group C (n:15) (p < 0.031, between groups A and C). Disappearance of the cysts from the stools on day 15 was 80% in group B, 72.2% in group A, and 26.6% in group C (p = 0.011, between group B and group C; p = 0.013, between group A and group C). At the end of the first month after inclusion, clinical cure rate was 94.4% in group A and 73.3% in group B (p = 0.11). Parasitological cure rate for B. hominis was very comparable between both groups (94.4% vs. 93.3%, p = 0.43). Metronidazole or S. boulardii has potential beneficial effects in B. hominis infection (symptoms, presence of parasites). These findings challenge the actual guidelines. PMID:20922415

  11. Virulence, persistence and dissemination of Mycoplasma bovis.

    PubMed

    Bürki, Sibylle; Frey, Joachim; Pilo, Paola

    2015-08-31

    Bovine mycoplasmosis due to Mycoplasma bovis causes several important bovine diseases such as pneumonia, mastitis, arthritis, otitis, genital disorders or keratoconjunctivitis. Variable surface lipoproteins, adhesion, invasion of host cells, modulation of the host immune system, biofilm formation and the release of secondary metabolites like hydrogen peroxide, as well as synergistic infections with other bacterial or viral pathogens are among the more significantly studied characteristics of the bacterium. The aim of this review is to summarize the current knowledge regarding the virulence of M. bovis and additionally, factors contributing to the dissemination and persistence of this pathogen in the bovine host will be discussed. PMID:25824130

  12. Catalase Enhances Growth and Biofilm Production of Mycoplasma pneumoniae.

    PubMed

    Simmons, Warren L; Dybvig, Kevin

    2015-08-01

    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

  13. [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. PMID:3090766

  14. Rapid detection of Mycoplasma synoviae by loop-mediated isothermal amplification.

    PubMed

    Kursa, Olimpia; Wo?niakowski, Grzegorz; Tomczyk, Grzegorz; Sawicka, Anna; Minta, Zenon

    2015-03-01

    Mycoplasma synoviae (MS) remains a serious concern in production of poultry and affects world production of chickens and turkeys. Loop-mediated isothermal amplification (LAMP) of DNA has been recently used for the identification of different economically important avian pathogens. The aim of this study was to develop LAMP for simple and inexpensive detection of MS strains in poultry using specifically designed primers targeting hemagglutin A (vlh) gene. The assay was conducted in a water bath for 1 h at 63 °C. The results were visualized after addition of SYBR Green(®) fluorescent dye. LAMP was specific exclusively for MS without cross-reactivity with other Mycoplasma species. The sensitivity of LAMP was determined as 10(-1) CFU/ml and was 1,000 times higher than MS-specific polymerase chain reaction. LAMP assay was conducted on 18 MS field strains to ensure its reliability and usefulness. This is the first report on LAMP development and application for the rapid detection of MS isolated from chickens. This simple method may be applied by diagnostic laboratories without access to expensive equipment. PMID:25413672

  15. New insights on the biology of swine respiratory tract mycoplasmas from a comparative genome analysis

    PubMed Central

    2013-01-01

    Background Mycoplasma hyopneumoniae, Mycoplasma flocculare and Mycoplasma hyorhinis live in swine respiratory tracts. M. flocculare, a commensal bacterium, is genetically closely related to M. hyopneumoniae, the causative agent of enzootic porcine pneumonia. M. hyorhinis is also pathogenic, causing polyserositis and arthritis. In this work, we present the genome sequences of M. flocculare and M. hyopneumoniae strain 7422, and we compare these genomes with the genomes of other M. hyoponeumoniae strain and to the a M. hyorhinis genome. These analyses were performed to identify possible characteristics that may help to explain the different behaviors of these species in swine respiratory tracts. Results The overall genome organization of three species was analyzed, revealing that the ORF clusters (OCs) differ considerably and that inversions and rearrangements are common. Although M. flocculare and M. hyopneumoniae display a high degree of similarity with respect to the gene content, only some genomic regions display considerable synteny. Genes encoding proteins that may be involved in host-cell adhesion in M. hyopneumoniae and M. flocculare display differences in genomic structure and organization. Some genes encoding adhesins of the P97 family are absent in M. flocculare and some contain sequence differences or lack of domains that are considered to be important for adhesion to host cells. The phylogenetic relationship of the three species was confirmed by a phylogenomic approach. The set of genes involved in metabolism, especially in the uptake of precursors for nucleic acids synthesis and nucleotide metabolism, display some differences in copy number and the presence/absence in the three species. Conclusions The comparative analyses of three mycoplasma species that inhabit the swine respiratory tract facilitated the identification of some characteristics that may be related to their different behaviors. M. hyopneumoniae and M. flocculare display many differences that may help to explain why one species is pathogenic and the other is considered to be commensal. However, it was not possible to identify specific virulence determinant factors that could explain the differences in the pathogenicity of the analyzed species. The M. hyorhinis genome contains differences in some components involved in metabolism and evasion of the host’s immune system that may contribute to its growth aggressiveness. Several horizontal gene transfer events were identified. The phylogenomic analysis places M. hyopneumoniae, M. flocculare and M. hyorhinis in the hyopneumoniae clade. PMID:23497205

  16. The linear chromosome of the plant-pathogenic mycoplasma 'Candidatus Phytoplasma mali'

    PubMed Central

    Kube, Michael; Schneider, Bernd; Kuhl, Heiner; Dandekar, Thomas; Heitmann, Katja; Migdoll, Alexander M; Reinhardt, Richard; Seemüller, Erich

    2008-01-01

    Background Phytoplasmas are insect-transmitted, uncultivable bacterial plant pathogens that cause diseases in hundreds of economically important plants. They represent a monophyletic group within the class Mollicutes (trivial name mycoplasmas) and are characterized by a small genome with a low GC content, and the lack of a firm cell wall. All mycoplasmas, including strains of 'Candidatus (Ca.) Phytoplasma asteris' and 'Ca. P. australiense', examined so far have circular chromosomes, as is the case for almost all walled bacteria. Results Our work has shown that 'Ca. Phytoplasma mali', the causative agent of apple proliferation disease, has a linear chromosome. Linear chromosomes were also identified in the closely related provisional species 'Ca. P. pyri' and 'Ca. P. prunorum'. The chromosome of 'Ca. P. mali' strain AT is 601,943 bp in size and has a GC content of 21.4%. The chromosome is further characterized by large terminal inverted repeats and covalently closed hairpin ends. Analysis of the protein-coding genes revealed that glycolysis, the major energy-yielding pathway supposed for 'Ca. P. asteris', is incomplete in 'Ca. P. mali'. Due to the apparent lack of other metabolic pathways present in mycoplasmas, it is proposed that maltose and malate are utilized as carbon and energy sources. However, complete ATP-yielding pathways were not identified. 'Ca. P. mali' also differs from 'Ca. P. asteris' by a smaller genome, a lower GC content, a lower number of paralogous genes, fewer insertions of potential mobile DNA elements, and a strongly reduced number of ABC transporters for amino acids. In contrast, 'Ca. P. mali' has an extended set of genes for homologous recombination, excision repair and SOS response than 'Ca. P. asteris'. Conclusion The small linear chromosome with large terminal inverted repeats and covalently closed hairpin ends, the extremely low GC content and the limited metabolic capabilities reflect unique features of 'Ca. P. mali', not only within phytoplasmas, but all mycoplasmas. It is expected that the genome information obtained here will contribute to a better understanding of the reduced metabolism of phytoplasmas, their fastidious nutrition requirements that prevented axenic cultivation, and the mechanisms involved in pathogenicity. PMID:18582369

  17. Elucidation of the life cycle of the intestinal protozoan Blastocystis hominis

    Microsoft Academic Search

    M. Singh; K. Suresh; L. C. Ho; G. C. Ng; E. H. Yap

    1995-01-01

    This paper elucidates the status of the different morphological forms ofBlastocystis and reports the existence of thin-and thick-walled cysts inB. hominis on the basis of current experimental evidence. It is suggested that the thin-walled cysts are autoinfectious, leading to multiplication of the organism in the intestinal tract. The thick-walled cysts are responsible for external transmission via the faecal-oral route. A

  18. [Observations on Blastocystis hominis and Cyclospora cayetanensis in routine parasitological examinations].

    PubMed

    Alarcón, Ruth Semira Rodriguez; Amato Neto, Vicente; Gakiya, Erika; Bezerra, Rita Cristina

    2007-01-01

    We report some observations made from routine parasitological examinations on feces. The methods of Faust et al. and of spontaneous sedimentation in water are not enough to identify Blastocystis hominis. Significant percentage presence of this protozoan was found, especially when staining with iron hematoxylin was performed. Cyclospora cayetanensis was found in 0.7% of the cases, which suggests that this parasite should also routinely be investigated by appropriate techniques. PMID:17568902

  19. 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

  20. Complete development and multiplication of Cryptosporidium hominis in cell-free culture.

    PubMed

    Hijjawi, Nawal; Estcourt, Annika; Yang, Rongchang; Monis, Paul; Ryan, Una

    2010-04-19

    The present study reports for the first time the completion of the life cycle of Cryptosporidium hominis in cell-free culture and multiplication of the parasite via qPCR. Individual life-cycle stages were characterised using Cryptosporidium-specific antibody staining (Sporo-Glo) and fluorescent in situ hybridisation (FISH) staining on cultures inoculated with excysted oocysts and purified sporozoites. In both cultures, C. hominis successfully proliferated and completed its life cycle, however development in cultures inoculated with purified sporozoites lagged behind cultures inoculated with excysted oocysts. Some novel findings of the study include the visualisation of pairing and multiple associations between various developmental stages in a process similar to syzygy and the formation of Cryptosporidium stages (trophozoites and meronts) inside the oocysts without excystation. qPCR analysis revealed a 5-6-fold amplification of parasite DNA. Future studies are required to improve the amplification of the parasite. The present study confirms the suitability of this culturing model to support the growth and proliferation of C. hominis (which unlike C. parvum, cannot be readily cultured in small animal models) and will greatly assist in our understanding of the developmental biology of Cryptosporidium, its position within the Apicomplexa and its relationship to gregarine protozoa. PMID:20092948

  1. Rhamnose Biosynthesis in Mycoplasmas Requires Precursor Glycans Larger than Monosaccharide

    PubMed Central

    Jordan, David S.; Daubenspeck, James M.; Dybvig, Kevin

    2013-01-01

    Summary Despite the apparent absence of genes coding for the known pathways for biosynthesis, the monosaccharide rhamnose was detected in the D configuration in Mycoplasma pneumoniae and Mycoplasma pulmonis, and in both the D and L configurations in Mycoplasma arthritidis. Surprisingly, the monosaccharide glucose was not a precursor for rhamnose biosynthesis and was not incorporated at detectable levels in glucose-containing polysaccharides or glycoconjugates. In contrast, carbon atoms from starch, a polymer of glucose, were incorporated into rhamnose in each of the three species examined. When grown in a serum-free medium supplemented with starch, M. arthritidis synthesized higher levels of rhamnose, with a shift in the relative amounts of the D and L configurations. Our findings suggest the presence of a novel pathway for rhamnose synthesis that is widespread in the genus Mycoplasma. PMID:23826905

  2. Mycoplasmas: Sophisticated, Reemerging, and Burdened by Their Notoriety

    Microsoft Academic Search

    Joel B. Baseman; Joseph G. Tully; Thomas Henry Huxley

    1997-01-01

    Mycoplasmas are most unusual self-replicating bacteria, possessing very small genomes, lacking cell wall components, requiring cholesterol for membrane function and growth, using UGA codon for tryptophan, passing through \\

  3. 21 CFR 864.2360 - Mycoplasma detection media and components.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Cell And Tissue Culture Products § 864.2360 Mycoplasma detection media and components. (a)...

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

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Cell And Tissue Culture Products § 864.2360 Mycoplasma detection media and components. (a)...

  5. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...Mycoplasma pneumoniae range from inapparent infection to mild or severe upper respiratory disease, ear infection, and bronchial pneumonia. (b) Classification. Class I (general controls). The device is exempt from the premarket notification...

  6. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...Mycoplasma pneumoniae range from inapparent infection to mild or severe upper respiratory disease, ear infection, and bronchial pneumonia. (b) Classification. Class I (general controls). The device is exempt from the premarket notification...

  7. Variable expression and geographic distribution of Mycoplasma agalactiae surface epitopes demonstrated with monoclonal antibodies.

    PubMed

    Bergonier, D; De Simone, F; Russo, P; Solsona, M; Lambert, M; Poumarat, F

    1996-10-01

    Species-specific monoclonal antibodies (MAbs) were developed against Mycoplasma agalactiae reference strain PG2 and French isolate P89 to study the in vitro expression of surface epitopes and to probe the antigenic profiles of 245 field isolates originating from 10 different countries. Colony immunostaining with MAbs on clonal lineage showed that 4 out of 9 species-specific epitopes exhibited a high rate of variation, demonstrating that M. agalactiae possesses a capacity for phenotypic diversification of its surface antigenicity. The emphasis was on dot immunobinding screening of the field isolates with MAbs recognizing permanently expressed epitopes. Eight different profiles could be defined. Great differences in epitope conservation were demonstrated with some area-specific strains completely lacking certain specific determinants. These results indicate that the antigenic variability of M. agalactiae relies not only upon surface switching mechanisms but also upon true epitope differences, partially related to the geographic origin of the isolates. PMID:8837468

  8. Blastocystis hominis infection in a post-cardiotomy patient on extracorporeal membrane oxygenation support: A case report and literature review

    PubMed Central

    Chen, Chih-Hsuan; Sun, Hsin-Yun; Chien, Hsiung-Fei; Lai, Hong-Shiee; Chou, Nai-Kuan

    2014-01-01

    INTRODUCTION Opportunistic pathogens can cause severe damage leading to irreversible complications in immune-compromised patients. Here we describe a patient who sustained Blastocystis hominis infection resulting in severe sepsis while on extracorporeal membrane oxygenation (ECMO) support, and the course of treatment taken to treat him. PRESENTATION OF CASE Our case, a 34-year-old Filipino man, was hospitalized for valvular disease and received valve replacements. ECMO and an intra-aortic balloon pump (IABP) were implemented when the patient developed progressive heart failure after cardiac surgery. Unfortunately, the patient suffered from sepsis with persistent fever and diarrhea, and subsequent examinations indicated the patient was infected by B. hominis. After adequate administration of the antibiotic metronidazole, the patient's symptoms subsided and he was discharged. DISCUSSION Blastocystis hominis is a unicellular protozoa commonly found in the intestinal tract, and the prevalence of B. hominis is 1.5–10% in developed countries and 30–50% in developing countries. The patient needed the support of ECMO and IABP, was immunocompromised to a certain extent; B. hominis can be a harmful opportunistic pathogen for them and lead to severe irreversible complications such as death. CONCLUSION This is the first published article showing that the opportunistic pathogen, B. hominis, can cause severe infection in patients on ECMO support, a result that should be kept in mind when patients come from a place with a high prevalence of B. hominis. The prophylactic medication should be administered routinely when patients live in the region and extracorporeal life-support is used. PMID:25160800

  9. Role of Binding in Mycoplasma mobile and Mycoplasma pneumoniae Gliding Analyzed through Inhibition by Synthesized Sialylated Compounds

    PubMed Central

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

    2013-01-01

    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. PMID:23123913

  10. Role of the GapA and CrmA Cytadhesins of Mycoplasma gallisepticum in Promoting Virulence and Host Colonization

    PubMed Central

    Indiková, Ivana; Much, Peter; Stipkovits, László; Siebert-Gulle, Karin; Citti, Christine

    2013-01-01

    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. PMID:23460514

  11. 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. PMID:23778631

  12. Mycoplasma mycoides subsp. capri associated with goat respiratory disease and high flock mortality

    PubMed Central

    Hernandez, Laura; St-Jacques, Marcel; Ontiveros, Lourdes; Acosta, Jorge; Handel, Katherine

    2006-01-01

    Abstract A high mortality outbreak of respiratory mycoplasmosis occurred in goats in Mexico. The clinicopathologic presentation resembled contagious caprine pleuropneumonia caused by Mycoplasma capricolum subspecies capripneumoniae. By using a battery of polymerase chain reaction assays, the mycoplasma associated with this outbreak was identified as Mycoplasma mycoides subsp. capri. PMID:16642877

  13. Inhibition of Mycoplasma pneumoniae growth by FDA-approved anticancer and antiviral nucleoside and nucleobase analogs

    PubMed Central

    2013-01-01

    Background Mycoplasma pneumoniae (Mpn) is a human pathogen that causes acute and chronic respiratory diseases and has been linked to many extrapulmonary diseases. Due to the lack of cell wall, Mpn is resistant to antibiotics targeting cell wall synthesis such as penicillin. During the last 10 years macrolide-resistant Mpn strains have been frequently reported in Asian countries and have been spreading to Europe and the United States. Therefore, new antibiotics are needed. In this study, 30 FDA-approved anticancer or antiviral drugs were screened for inhibitory effects on Mpn growth and selected analogs were further characterized by inhibition of target enzymes and metabolism of radiolabeled substrates. Results Sixteen drugs showed varying inhibitory effects and seven showed strong inhibition of Mpn growth. The anticancer drug 6-thioguanine had a MIC (minimum inhibitory concentration required to cause 90% of growth inhibition) value of 0.20 ?g ml-1, whereas trifluorothymidine, gemcitabine and dipyridamole had MIC values of approximately 2 ?g ml-1. In wild type Mpn culture the presence of 6-thioguanine and dipyridamole strongly inhibited the uptake and metabolism of hypoxanthine and guanine while gemcitabine inhibited the uptake and metabolism of all nucleobases and thymidine. Trifluorothymidine and 5-fluorodeoxyuridine, however, stimulated the uptake and incorporation of radiolabeled thymidine and this stimulation was due to induction of thymidine kinase activity. Furthermore, Mpn hypoxanthine guanine phosphoribosyl transferase (HPRT) was cloned, expressed, and characterized. The 6-thioguanine, but not other purine analogs, strongly inhibited HPRT, which may in part explain the observed growth inhibition. Trifluorothymidine and 5-fluorodeoxyuridine were shown to be good substrates and inhibitors for thymidine kinase from human and Mycoplasma sources. Conclusion We have shown that several anticancer and antiviral nucleoside and nucleobase analogs are potent inhibitors of Mpn growth and that the mechanism of inhibition are most likely due to inhibition of enzymes in the nucleotide biosynthesis pathway and nucleoside transporter. Our results suggest that enzymes in Mycoplasma nucleotide biosynthesis are potential targets for future design of antibiotics against Mycoplasma infection. PMID:23919755

  14. Mycoplasma genitalium: a cause of male urethritis?

    PubMed Central

    Jensen, J S; Orsum, R; Dohn, B; Uldum, S; Worm, A M; Lind, K

    1993-01-01

    BACKGROUND--Male urethritis may be caused by mycoplasmas. Since Mycoplasma genitalium has previously been isolated from the urethra of two men with non-gonococcal urethritis (NGU), it was the aim of the study further to elucidate its role by measuring the prevalence of this organism in men with NGU. MATERIAL AND METHODS--The polymerase chain reaction was used. Two different sequences of the gene coding for the main adhesin MgPa were amplified. Urethral, rectal, and throat samples from 99 male sexually transmitted disease (STD) patients with and without urethritis were studied. RESULTS--M genitalium DNA was demonstrated in 17/99 (17%) of the urethral swabs, but in none of the rectal and throat swabs. Significantly more patients with urethritis (13/52) were positive for M genitalium DNA than were patients without urethritis (4/47) (p < 0.03). In those with urethritis M genitalium DNA was found more often in Chlamydia trachomatis negative NGU (12/34) than in those with chlamydial NGU (1/14) (p = 0.05). Attempts to culture M genitalium from the PCR positive specimens were unsuccessful. CONCLUSION--M genitalium DNA was found significantly more often in male STD patients with non-chlamydial NGU than in men with chlamydial urethritis (p = 0.05) and in men without urethritis (p = 0.003), suggesting that M genitalium may be a cause of NGU. M genitalium DNA was not demonstrated in any of the throat or rectal swabsindicating that the urogenital tract is probably the primary site of infection or colonisation of this species. PMID:7721285

  15. Characterization of MGC2, a Mycoplasma gallisepticum Cytadhesin with Homology to the Mycoplasma pneumoniae 30-Kilodalton Protein P30 and Mycoplasma genitalium P32†

    PubMed Central

    Hnatow, Linda L.; Keeler, Calvin L.; Tessmer, Laura L.; Czymmek, Kirk; Dohms, John E.

    1998-01-01

    A second cytadhesin-like protein, MGC2, was identified in the avian respiratory pathogen Mycoplasma gallisepticum. The 912-nucleotide mgc2 gene encodes a 32.6-kDa protein with 40.9 and 31.4% identity with the M. pneumoniae P30 and M. genitalium P32 cytadhesins, respectively. Functional studies with reverse transcription-PCR, immunoblotting, double-sided immunogold labeling, and attachment inhibition assays demonstrated homology to the human mycoplasmal P30 and P32 cytadhesins. These findings suggest that there is a family of cytadhesin genes conserved among pathogenic mycoplasmas infecting widely divergent hosts. PMID:9632619

  16. Characterization of MGC2, a Mycoplasma gallisepticum cytadhesin with homology to the Mycoplasma pneumoniae 30-kilodalton protein P30 and Mycoplasma genitalium P32.

    PubMed

    Hnatow, L L; Keeler, C L; Tessmer, L L; Czymmek, K; Dohms, J E

    1998-07-01

    A second cytadhesin-like protein, MGC2, was identified in the avian respiratory pathogen Mycoplasma gallisepticum. The 912-nucleotide mgc2 gene encodes a 32.6-kDa protein with 40.9 and 31.4% identity with the M. pneumoniae P30 and M. genitalium P32 cytadhesins, respectively. Functional studies with reverse transcription-PCR, immunoblotting, double-sided immunogold labeling, and attachment inhibition assays demonstrated homology to the human mycoplasmal P30 and P32 cytadhesins. These findings suggest that there is a family of cytadhesin genes conserved among pathogenic mycoplasmas infecting widely divergent hosts. PMID:9632619

  17. Loop-mediated isothermal amplification for rapid and convenient detection of Mycoplasma hyopneumoniae.

    PubMed

    Li, Jiahe; Minion, F Chris; Petersen, Andrew C; Jiang, Fei; Yang, Sheng; Guo, Panpan; Li, Jinxiang; Wu, Wenxue

    2013-04-01

    Loop-mediated isothermal amplification (LAMP), a novel method of gene amplification, was employed in this study for detecting Mycoplasma hyopneumoniae in the respiratory tract or lungs of swine. The pathogen can be detected in LAMP reactions containing as few as 10 fg purified target DNA (10 copies of M. hyopneumoniae genome) within 30 min, which was comparable to real-time PCR. After 30-min reaction at 63 °C, the addition of a certain amount of dye (SYBR Green I and hydroxyl naphthol blue at a proper ratio) into the LAMP reaction system makes the results easily determined as positive or negative by visual inspection. In addition, the LAMP was able to distinguish between M. hyopneumoniae and other closely-related mycoplasma strains, indicating a high degree of specificity. The LAMP assay was more simple and cheap, since the reaction could be completed under isothermal conditions and less laboratorial infrastructure are required. And, it was proven reliable for M. hyopneumoniae diagnosis of nasal swab and lung samples from the field. PMID:23184577

  18. cDNA and genomic cloning and expression of the P48 monocytic differentiation/activation factor, a Mycoplasma fermentans gene product.

    PubMed Central

    Hall, R E; Agarwal, S; Kestler, D P; Cobb, J A; Goldstein, K M; Chang, N S

    1996-01-01

    P48 is a 48 kDa monocytic differentiation/activation factor previously purified from the conditioned medium of the Reh human pre-B cell leukaemia cell line. It induces growth arrest and differentiation of HL-60 human promyelocytic leukaemia cells along the monocytic pathway and the production of the cytokines interleukin 1, tumour necrosis factor-alpha and interleukin 6 in human monocytes and monocytic cell lines. The cDNA for P48 was cloned from Reh cellular RNA using 3' reverse amplification of cDNA ends. Southern blot probing with P48 cDNA revealed hybridization with DNA from Reh and Molt-4 cells, but not with DNA from human peripheral blood mononuclear cells. Subsequent studies using PCR and Southern analysis revealed P48 sequences in DNA isolated from Mycoplasma fermentans but not M. hominis, M.iowae, M.synoviae or M.lypophilum. Although initial studies using Mycoplasma culture and hybridization techniques had failed to reveal Mycoplasma infection in our Reh and Molt-4 cell lines, subsequent PCR studies using Mycoplasma genus-specific rRNA primrs revealed Mycoplasma sequences in these cell lines. Using the P48 cDNA probe, we isolated a genomic clone from M. fermentans DNA which was found to be 98.5% identical with the P48 cDNA clone, and the deduced amino acid sequence agreed with N-terminal microsequencing data for P48 protein purified from the Reh cell line conditioned medium. The 5' end of the gene has a number of consensus sequences characteristic of prokaryotic genes, and the deduced amino acid sequence has a number of features suggesting that P48 is a lipoprotein. The P48 cDNA was expressed in pMAL in Escherichia coli, and the 60 kDa expressed fusion protein was found to react with anti-P48 antibodies on Western blots. This is consistent with a pMAL fusion protein representing the sum of the 42 kDa maltose-binding protein and 18 kDa of P48 recombinant protein, suggesting that native P48 has significant post-translational modification. Consistent with this, Northern blot studies revealed a single 1 kb transcript. The recombinant fusion protein was found to possess anti-proliferative activity against HL-60 cells, and antibodies against recombinant P48 were found to block the biological activity of native P48 isolated from conditioned medium. These studies demonstrate that P48, a molecule with immunomodulatory and haematopoietic differentiation activities, is derived from M. fermentans or a closely related species. P48 may be important in the pathophysiology of Mycoplasma infections and may be useful in dissecting the mechanisms involved in mammalian haematopoietic cell differentiation, immune function and cytokine biosynthesis. PMID:8921000

  19. Characterization and analysis of a stable serotype-associated membrane protein (P30) of Mycoplasma agalactiae.

    PubMed

    Fleury, B; Bergonier, D; Berthelot, X; Schlatter, Y; Frey, J; Vilei, E M

    2001-08-01

    The gene for a 30-kDa immunodominant antigen, P30, of Mycoplasma agalactiae was cloned from type strain PG2 and expressed in Escherichia coli. P30 is encoded on a monocistronic operon determined by two -10 boxes and a possible -35 region constituting the potential promoter, and a transcription termination site. The gene for the 266-amino-acid protein is preceded by a polypurine-rich region designed as the consensus sequence for a ribosome-binding site. Analysis of the amino acid sequence of P30 revealed the presence of a recognition site for a prokaryotic signal peptidase II at amino acid (aa) 24, indicating that P30 is a transmembrane protein. Moreover, Triton X-114 phase partitioning of M. agalactiae PG2 total antigen revealed that P30 is strongly hydrophobic and hence a possible membrane component. Immunoblot analysis using the monospecific polyclonal anti-P30-His serum indicated that P30 is specific to M. agalactiae. Furthermore, PCR amplification with specific primers for p30 and Southern blot analysis revealed the presence of the gene in all M. agalactiae strains tested and its absence in the other mycoplasma species. Among 27 strains of M. agalactiae studied, 20 strains belonging to the common serotypes A to D, including PG2, expressed P30 or part of it as detected by the monospecific polyclonal anti-P30 antibodies. The other seven strains belonging to the rarely isolated serotypes E to H were negative for P30. The p30 gene was sequenced in 15 strains of M. agalactiae, 10 of which expressed P30 or at least part of it and 5 of which did not express P30. The negative strains carried mutations in both -10 boxes of the promoters. These mutations seem to be responsible for the lack of P30 expression in these strains. Analysis of sera from sheep that were experimentally infected with M. agalactiae revealed that P30 induced a strong and persistent immune response which was still very high two months after infection. In contrast, currently used enzyme-linked immunosorbent assay serology gave only low titers. PMID:11473997

  20. Characterization and Analysis of a Stable Serotype-Associated Membrane Protein (P30) of Mycoplasma agalactiae

    PubMed Central

    Fleury, Bénédicte; Bergonier, Dominique; Berthelot, Xavier; Schlatter, Yvonne; Frey, Joachim; Vilei, Edy M.

    2001-01-01

    The gene for a 30-kDa immunodominant antigen, P30, of Mycoplasma agalactiae was cloned from type strain PG2 and expressed in Escherichia coli. P30 is encoded on a monocistronic operon determined by two ?10 boxes and a possible ?35 region constituting the potential promoter, and a transcription termination site. The gene for the 266-amino-acid protein is preceded by a polypurine-rich region designed as the consensus sequence for a ribosome-binding site. Analysis of the amino acid sequence of P30 revealed the presence of a recognition site for a prokaryotic signal peptidase II at amino acid (aa) 24, indicating that P30 is a transmembrane protein. Moreover, Triton X-114 phase partitioning of M. agalactiae PG2 total antigen revealed that P30 is strongly hydrophobic and hence a possible membrane component. Immunoblot analysis using the monospecific polyclonal anti-P30-His serum indicated that P30 is specific to M. agalactiae. Furthermore, PCR amplification with specific primers for p30 and Southern blot analysis revealed the presence of the gene in all M. agalactiae strains tested and its absence in the other mycoplasma species. Among 27 strains of M. agalactiae studied, 20 strains belonging to the common serotypes A to D, including PG2, expressed P30 or part of it as detected by the monospecific polyclonal anti-P30 antibodies. The other seven strains belonging to the rarely isolated serotypes E to H were negative for P30. The p30 gene was sequenced in 15 strains of M. agalactiae, 10 of which expressed P30 or at least part of it and 5 of which did not express P30. The negative strains carried mutations in both ?10 boxes of the promoters. These mutations seem to be responsible for the lack of P30 expression in these strains. Analysis of sera from sheep that were experimentally infected with M. agalactiae revealed that P30 induced a strong and persistent immune response which was still very high two months after infection. In contrast, currently used enzyme-linked immunosorbent assay serology gave only low titers. PMID:11473997

  1. Cover: The crystal structure of dihydrofolate reductase-thymidylate synthase (DHFR-TS) from Cryptosporidium hominis. From R. O'Neil, R. Lilien, B. R. Donald, R. Stroud, and A. Anderson. The crystal structure of dihydrofolate

    E-print Network

    Donald, Bruce Randall

    Cryptosporidium hominis. From R. O'Neil, R. Lilien, B. R. Donald, R. Stroud, and A. Anderson. The crystal structure of dihydrofolate reductase-thymidylate synthase from Cryptosporidium hominis reveals a novel Structure of Dihydrofolate Reductase-Thymidylate Synthase from Cryptosporidium hominis Reveals a Novel

  2. Detection of Mycoplasma genitalium by PCR Amplification of the 16S rRNA Gene

    PubMed Central

    Jensen, Jørgen Skov; Borre, Martin B.; Dohn, Birthe

    2003-01-01

    In order to develop a species-specific PCR for the detection of Mycoplasma genitalium, the sequence of 1,490 bases of the 16S rRNA gene was determined for M. genitalium G37 (type strain) and four Danish isolates of M. genitalium. The sequences of the four Danish strains, mutually different with respect to their MgPa gene, were 100% homologous, although they carried a single common base substitution compared to the type strain. Among members of the Mycoplasma pneumoniae phylogenetic cluster, M. genitalium showed the most-prominent homology to the 16S rRNA sequence of M. pneumoniae (98% homology). From regions showing the least homology to the M. pneumoniae 16S rRNA gene sequence, primers were chosen to amplify DNA from M. genitalium only. Two sets of primers were selected for their ability to detect <10 to 50 M. genitalium genome copies without cross-reactions with M. pneumoniae. The performance of these primers was compared to the performance of two pairs of primers amplifying parts of the MgPa adhesin gene; 1,030 randomly selected specimens submitted for Chlamydia trachomatis culture were screened with one of the 16S rRNA gene primer sets. A total of 41 specimens were found to be positive for this gene; 40 of these could be confirmed by one of the MgPa primer sets, whereas the other MgPa primer set detected only 21 positive specimens out of 40. These results indicate that estimates of the prevalence of M. genitalium in various populations using MgPa PCR primers could be incorrectly low if the PCR primers are located in variable regions of the MgPa gene. PMID:12517858

  3. Diagnosis and antimicrobial treatment of Mycoplasma genitalium infection: sobering thoughts.

    PubMed

    Taylor-Robinson, David

    2014-06-01

    The discovery of Mycoplasma genitalium in 1980-1981 eventually led to it becoming recognized as an important cause of non-gonococcal urethritis in men and also some genital tract diseases in women. Subsequent to the original isolation, further attempts failed over the next decade and reliable detection only became possible with the use of nucleic acid amplification techniques. Although tetracyclines, particularly doxycycline, were the first choice for treatment of non-gonococcal urethritis prior to the finding of M. genitalium, they were unsatisfactory for the treatment of M. genitalium-associated disease; the organisms were often not eliminated leading, for example, to chronic urethritis. However, the introduction of azithromycin, used as single-dose therapy for chlamydial infections, resulted in clearance of the mycoplasmal organisms from the genital tract and clinical recovery without the development of chronic disease. Nevertheless, such success was short-lived as M. genitalium, through mutation, began to develop resistance to azithromycin and M. genitalium mutants also began to circulate in some populations. In an attempt to counteract this, clinicians should give extended therapy, and in the future, microbiologists, using real-time PCRs, might be able to determine the existence of resistant strains in the local population and so advise on the most appropriate antibiotic. Other than azithromycin, there are a few options, moxifloxacin being one, although the recently reported resistance to this antibiotic is disturbing. In the short to medium term, combination therapy and/or the advent of a new antibiotic might abate the spread of resistance, but in the long term, there is potential for increasing prevalence of untreatable M. genitalium disease. In the future, attempts to develop a vaccine and, of equal importance, one to Chlamydia trachomatis, would not be out of place. PMID:24834454

  4. In vitro antimycoplasmal activity of oleuropein

    Microsoft Academic Search

    Pio Maria Furneri; Andreana Marino; Antonina Saija; Nicola Uccella; Giuseppe Bisignano

    2002-01-01

    The activity of oleuropein, a phenolic glycoside contained in olive oil, was investigated in vitro against Mycoplasma hominis, Mycoplasma fermentans, Mycoplasma pneumoniae and Mycoplasma pirum. Oleuropein inhibited mycoplasmas at concentrations from 20 to 320 mg\\/l. The MICs of oleuropein to M. pneumoniae, M. pirum, M. hominis and M. fermentans were 160, 320, 20 and 20 mg\\/l, respectively.

  5. Multilocus Sequence Typing of an Emerging Cryptosporidium hominis Subtype in the United States

    PubMed Central

    Tiao, Narry; Li, Na; Hlavsa, Michele

    2014-01-01

    The United States has experienced a substantial increase in the reported incidence of cryptosporidiosis since 2005. Accompanying this is the emergence of a new subtype of Cryptosporidium hominis based on variation at the 60-kDa glycoprotein (gp60) locus, IaA28R4, which has become a frequently identified subtype in both sporadic and outbreak-related cases. In this study, using multilocus sequence typing (MLST) at eight genetic loci, we characterized 62 specimens of IaA28R4 and 33 specimens of three other gp60 subtypes of C. hominis from four U.S. states with increased cryptosporidiosis incidences during the summer of 2008. Extensive genetic heterogeneity was seen within the gp60 subtype IaA28R4, but specimens from Ohio and southwestern states formed two distinct subpopulations, suggesting that there were at least two origins of IaA28R4 within the United States. Discordance in typing results was observed between gp60 and other genetic markers, especially DZ-HRGP, and this discordance was largely the result of genetic recombination within the gp60 subtype IaA28R4. The results of population genetic analyses supported the presence of two subpopulations of IaA28R4 and the occurrence of genetic recombination within this gp60 subtype. Thus, the IaA28R4 subtype at gp60 is likely a fitness marker for C. hominis, and genetic recombination is potentially a driving force in the emergence of the virulent IaA28R4 subtype in the United States. A rapid evolution of IaA28R4 was indicated by the observation of multiple MLST subtypes of IaA28R4 within two large outbreaks that lasted for extended periods and involved multiple swimming pools. PMID:24478483

  6. In vitro effect of some Egyptian herbal extracts against Blastocystis hominis.

    PubMed

    Abdel-Hafeez, Ekhlas H; Ahmad, Azza K; Andelgelil, Noha H; Abdellatif, Manal Z M; Kamal, Amany M; Mohamed, Rabie M

    2015-04-01

    Blastocystis hominis is an enteric parasite that inhabits the gastrointestinal tract of humans and many animals. This emerging parasite has a worldwide distribution. It is often identified as the most common eukaryotic organism reported in human fecal samples that showed a dramatic increase in recent years. Metronidazole is the main therapy for blastocystosis. However, frequent reports of treatment failure suggesting isolates resistance to metronidazole. This study determined the growth pattern and in vitro susceptibility of B. hominis to nitazoxanide (NTZ), garlic, ginger, onion and turmeric. Fecal samples positive for Blastocystis were collected from patients with irritable bowel syndrome (IBS), and processed for culture. Cultured samples were subjected to examination by light microscopy. Herbs' extracts was freshly prepared. Drug susceptibility assays was done using 0.1 mg/ml of NTZ, garlic, ginger, onion and turmeric. Effects assessed on parasite culture after 24 hr. and 48 hr. Cultured fecal samples of B. hominis have identified several forms of the organism; vacuolar, granular, amoeboid and cyst forms within 24 hr. Nitazoxanide treatment significantly (P < 0.001) lowered the parasite number after 48 hr. (mean, 337.5 ± 17.67) /ml. The reduction rate after 48 hr. compared to PBS was 93.33%. Ginger treatment significantly (P < 0.002) lowered the number of the parasite after 48 hr. (mean, 335 ± 7.07)/ml. Moreover, garlic treatment also significantly (P < 0.002) lowered the number of the parasite after 48 hr. (mean, 382.5 ± 10.60)/ml. The reduction rates after 48 hr. in these treated samples compared to PBS were 92.98% and 92.44% respectively. However, onion, and turmeric treatments insignificantly lowered the number of the parasite after 48 hr. (P < 0.15 & < 0.22 respectively). PMID:26012223

  7. Composition of a polysaccharide from mycoplasma (F-38) recognised by antibodies from goats with contagious pleuropneumonia.

    PubMed

    Rurangirwa, F R; McGuire, T C; Magnuson, N S; Kibor, A; Chema, S

    1987-03-01

    A polysaccharide was extracted by warm aqueous phenol from the F-38 strain of mycoplasma which causes contagious caprine pleuropneumonia (CCPP). After acid hydrolysis, the polysaccharide was found to be composed of the neutral sugars glucose, galactose, mannose and fucose and the amino sugars galactosamine and glucosamine. All the sugars were present in approximately equal quantities. Unmodified goat erythrocytes bound the polysaccharide readily and the sensitised cells reacted with antibodies in sera from goats with experimental or natural CCPP. The unique composition of the F-38 polysaccharide and the specific reactivity of polysaccharide-sensitised red cells with antibodies from CCPP infected animals suggests that the polysaccharide should be useful for identification of F-38 organisms and diagnosis of the disease. PMID:3589164

  8. Cloning, Expression, Purification, Crystallization and Preliminary X-ray Analysis of Mycoplasma Genitalium Protein MG289

    SciTech Connect

    Sippel, K.; Boehlein, S; Sakai, Y; Quirit, J; Agbandje-McKenna, M; Rosser, C; McKenna, R

    2009-01-01

    Mycoplasma genitalium is a human pathogen that is associated with nongonococcal urethritis in men and cervicitis in women. The cloning, expression, purification and crystallization of the protein MG289 from M. genitalium strain G37 are reported here. Crystals of MG289 diffracted X-rays to 2.8 {angstrom} resolution. The crystals belonged to the orthorhombic space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 49.7, b = 90.9, c = 176.1 {angstrom}. The diffraction data after processing had an overall R{sub merge} of 8.7%. The crystal structure of Cypl, the ortholog of MG289 from M. hyorhinis, has recently been determined, providing a reasonable phasing model; molecular replacement is currently under way.

  9. Mosaic genome of endobacteria in arbuscular mycorrhizal fungi: Transkingdom gene transfer in an ancient mycoplasma-fungus association.

    PubMed

    Torres-Cortés, Gloria; Ghignone, Stefano; Bonfante, Paola; Schüßler, Arthur

    2015-06-23

    For more than 450 million years, arbuscular mycorrhizal fungi (AMF) have formed intimate, mutualistic symbioses with the vast majority of land plants and are major drivers in almost all terrestrial ecosystems. The obligate plant-symbiotic AMF host additional symbionts, so-called Mollicutes-related endobacteria (MRE). To uncover putative functional roles of these widespread but yet enigmatic MRE, we sequenced the genome of DhMRE living in the AMF Dentiscutata heterogama. Multilocus phylogenetic analyses showed that MRE form a previously unidentified lineage sister to the hominis group of Mycoplasma species. DhMRE possesses a strongly reduced metabolic capacity with 55% of the proteins having unknown function, which reflects unique adaptations to an intracellular lifestyle. We found evidence for transkingdom gene transfer between MRE and their AMF host. At least 27 annotated DhMRE proteins show similarities to nuclear-encoded proteins of the AMF Rhizophagus irregularis, which itself lacks MRE. Nuclear-encoded homologs could moreover be identified for another AMF, Gigaspora margarita, and surprisingly, also the non-AMF Mortierella verticillata. Our data indicate a possible origin of the MRE-fungus association in ancestors of the Glomeromycota and Mucoromycotina. The DhMRE genome encodes an arsenal of putative regulatory proteins with eukaryotic-like domains, some of them encoded in putative genomic islands. MRE are highly interesting candidates to study the evolution and interactions between an ancient, obligate endosymbiotic prokaryote with its obligate plant-symbiotic fungal host. Our data moreover may be used for further targeted searches for ancient effector-like proteins that may be key components in the regulation of the arbuscular mycorrhiza symbiosis. PMID:25964335

  10. Comprehensive methylome characterization of Mycoplasma genitalium and Mycoplasma pneumoniae at single-base resolution.

    PubMed

    Lluch-Senar, Maria; Luong, Khai; Lloréns-Rico, Verónica; Delgado, Javier; Fang, Gang; Spittle, Kristi; Clark, Tyson A; Schadt, Eric; Turner, Stephen W; Korlach, Jonas; Serrano, Luis

    2013-01-01

    In the bacterial world, methylation is most commonly associated with restriction-modification systems that provide a defense mechanism against invading foreign genomes. In addition, it is known that methylation plays functionally important roles, including timing of DNA replication, chromosome partitioning, DNA repair, and regulation of gene expression. However, full DNA methylome analyses are scarce due to a lack of a simple methodology for rapid and sensitive detection of common epigenetic marks (ie N(6)-methyladenine (6 mA) and N(4)-methylcytosine (4 mC)), in these organisms. Here, we use Single-Molecule Real-Time (SMRT) sequencing to determine the methylomes of two related human pathogen species, Mycoplasma genitalium G-37 and Mycoplasma pneumoniae M129, with single-base resolution. Our analysis identified two new methylation motifs not previously described in bacteria: a widespread 6 mA methylation motif common to both bacteria (5'-CTAT-3'), as well as a more complex Type I m6A sequence motif in M. pneumoniae (5'-GAN(7)TAY-3'/3'-CTN(7)ATR-5'). We identify the methyltransferase responsible for the common motif and suggest the one involved in M. pneumoniae only. Analysis of the distribution of methylation sites across the genome of M. pneumoniae suggests a potential role for methylation in regulating the cell cycle, as well as in regulation of gene expression. To our knowledge, this is one of the first direct methylome profiling studies with single-base resolution from a bacterial organism. PMID:23300489

  11. Molecular epidemiologic investigations of Mycoplasma gallisepticum conjunctivitis in songbirds by random amplified polymorphic DNA analyses.

    PubMed

    Ley, D H; Berkhoff, J E; Levisohn, S

    1997-01-01

    An ongoing outbreak of conjunctivitis in free-ranging house finches (Carpodacus mexicanus) began in 1994 in the eastern United States. Bacterial organisms identified as Mycoplasma gallisepticum (MG) were isolated from lesions of infected birds. MG was also isolated from a blue jay (Cyanocitta cristata) that contracted conjunctivitis after being housed in a cage previously occupied by house finches with conjunctivitis, and from free-ranging American goldfinches (Carduelis tristis) in North Carolina in 1996. To investigate the molecular epidemiology of this outbreak, we produced DNA fingerprints of MG isolates by random amplification of polymorphic DNA (RAPD). We compared MG isolates from songbirds examined from 1994 through 1996 in 11 states, representing three host species, with vaccine and reference strains and with contemporary MG isolates from commercial poultry. All MG isolates from songbirds had RAPD banding patterns identical to each other but different from other strains and isolates tested. These results indicate that the outbreak of MG in songbirds is caused by the same strain, which suggests a single source; the outbreak is not caused by the vaccine or reference strains analyzed; and MG infection has not been shared between songbirds and commercial poultry. PMID:9284386

  12. Cross-Genome Comparisons of Newly Identified Domains in Mycoplasma gallisepticum and Domain Architectures with Other Mycoplasma species

    PubMed Central

    Chilamakuri, Chandra Sekhar Reddy; Joshi, Adwait; Rani, Sane Sudha; Offmann, Bernard; Sowdhamini, R.

    2011-01-01

    Accurate functional annotation of protein sequences is hampered by important factors such as the failure of sequence search methods to identify relationships and the inherent diversity in function of proteins related at low sequence similarities. Earlier, we had employed intermediate sequence search approach to establish new domain relationships in the unassigned regions of gene products at the whole genome level by taking Mycoplasma gallisepticum as a specific example and established new domain relationships. In this paper, we report a detailed comparison of the conservation status of the domain and domain architectures of the gene products that bear our newly predicted domains amongst 14 other Mycoplasma genomes and reported the probable implications for the organisms. Some of the domain associations, observed in Mycoplasma that afflict humans and other non-human primates, are involved in regulation of solute transport and DNA binding suggesting specific modes of host-pathogen interactions. PMID:21860605

  13. First steps towards the genetic manipulation of Mycoplasma agalactiae and Mycoplasma bovis using the transposon Tn4001mod.

    PubMed

    Chopra-Dewasthaly, Rohini; Zimmermann, Martina; Rosengarten, Renate; Citti, Christine

    2005-01-01

    Mycoplasma agalactiae and M. bovis rank amongst the most serious pathogenic mycoplasmas infecting small ruminants and cattle, respectively. Despite considerable advances made in Mycoplasma molecular genetics in the past decade, there is still a complete lack of genetic tools to assess the pathogenic mechanisms of these two species. Studies were undertaken to develop a genetic system for the analysis of potential virulence factors of these pathogens. Transposon Tn4001mod was successfully introduced into various chromosomal sites of M. agalactiae and M. bovis with an optimal frequency of 10(-6) per viable colony-forming unit (CFU). This is the first report that demonstrates the amenability of these agents to transformation and to genetic manipulation. Furthermore, Tn4001 is implicated as the first potential genetic tool available for these ruminant pathogens. PMID:15715173

  14. Smooth muscle antibodies in Mycoplasma pneumoniae infection.

    PubMed Central

    Biberfeld, G; Sterner, G

    1976-01-01

    Paired sera from forty-five cases of Mycoplasma pneumoniae (MP) infection associated with acute lower respiratory tract illness were examined by immunofluorescence for antibodies to smooth muscle. Twenty-five (56%) of these cases had smooth muscle antibodies (SMA) of IgM class. A significant (greater than or equal to 4-fold) increase in titre of these antibodies was demonstrated in fifteen of thirty-five patients with a significant rise in titre of MP antibodies. SMA of IgG class occurred in eleven of forty-five cases (24%), but a 4-fold rise in antibody titre was found only in two cases. Three of forty-five sera (7%) from healthy donors contained SMA of IgM class and eight sera (18%) SMA of IgG class. MP antigen did not absorb SMA. Liver tests were performed in twenty-nine patients. In eighteen patients SGPT values were moderately or slightly elevated. There was no correlation between the occurrence of increased levels of transaminases and the presence of SMA in serum. In a patient with active chronic hepatitis, who had had a high titre of SMA exclusively of IgG class for 2 years, SMA of IgM class appeared transiently in association with an acute respiratory illness due to MP. PMID:1084242

  15. Uptake of fatty acids by Mycoplasma capricolum.

    PubMed Central

    Dahl, J

    1988-01-01

    The energy requirements for fatty acid uptake by Mycoplasma capricolum were studied. Fatty acid transport and esterification to phospholipid appeared to be tightly coupled, since there was little intracellular accumulation of free fatty acid. Uptake was blocked by iodoacetate, n-ethylmaleimide, and p-chloromercuribenzoate. Glucose, glycerol, and potassium ions were necessary for fatty acid uptake by whole cells. A reduction in uptake was observed in cells treated with valinomycin or dicyclohexylcarbodiimide. The effect of temperature on the rate of oleate uptake showed a discontinuity at 24 degrees C. Above 24 degrees C an energy of activation of 4.6 kcal (ca. 19.2 kJ)/mol was obtained. The data suggest that uptake of fatty acid by M. capricolum is an energy-linked, protein-mediated process. A membrane-bound enzyme activity that catalyzed the synthesis of fatty acyl-hydroxamate was demonstrated. This activity was virtually independent or only marginally dependent on coenzyme A, depending on the assay system, but was stimulated approximately twofold by ATP. PMID:2834319

  16. Comparative analysis of the Mycoplasma capricolum subsp. capricolum GM508D genome reveals subrogation of phase-variable contingency genes and a novel integrated genetic element.

    PubMed

    Calcutt, Michael J; Foecking, Mark F

    2015-08-01

    Mycoplasma capricolum subspecies capricolum is both a pathogen of small ruminants and a model recipient organism for gene transplantation and synthetic biology. With the availability of the complete genome of the type strain California kid (released in 2005), a draft genome of strain GM508D was determined to investigate genomic variation in this subspecies. Differences in mobile genetic element location and complement, catabolic pathway genes, contingency loci, surface antigen genes and type II restriction-modification systems highlight the plasticity and diversity within this taxon. PMID:26040761

  17. Pre-AIDS Era Isolates of Pneumocystis carinii f. sp. hominis: High Genotypic Similarity with Contemporary Isolates

    PubMed Central

    Tsolaki, Anthony G.; Beckers, Pieter; Wakefield, Ann E.

    1998-01-01

    Isolates of Pneumocystis carinii f. sp. hominis were examined from six individuals who died of P. carinii pneumonia between 1968 and 1981 and who had underlying immunodeficiencies which were not due to human immunodeficiency virus infection. DNA sequence variation was analyzed in the genes encoding the mitochondrial large subunit rRNA (mt LSU rRNA), the internal transcribed spacer (ITS) regions of the nuclear rRNA, the arom locus, and the mitochondrial small subunit rRNA. No major variations were observed when these isolates were compared to isolates from HIV-infected individuals. A small number of minor differences were detected. A new position at which variation occurred in the mt LSU rRNA was observed in one sample. Three new ITS sequence types were identified. A total of nine different ITS sequence types were found in the six samples. Mixed infection with different ITS sequence types of P. carinii f. sp. hominis was observed in four of the six samples. The ITS locus was the most informative of the four loci for distinguishing among the isolates of P. carinii f. sp. hominis. The data suggest that isolates of P. carinii f. sp. hominis from before the AIDS pandemic are genetically very similar to those currently found in HIV-infected individuals. PMID:9431927

  18. The association of Blastocystis hominis and Endolimax nana with diarrheal stools in Zambian school-age children.

    PubMed

    Graczyk, Thaddeus K; Shiff, Clive K; Tamang, Leena; Munsaka, Fair; Beitin, Anna M; Moss, William J

    2005-12-01

    To determine the prevalence of endoparasites and their association with diarrhea, a survey was conducted in the Southern Province of Zambia that used conventional and molecular techniques applied to stool and urine samples from school-age children (n = 93). Almost half of the stools (49.5%) were diarrhetic. The overall prevalence of Endolimax nana, Schistosoma haematobium, Blastocystis hominis, Giardia lamblia, Cryptosporidium parvum, Encephalitozoon intestinalis, and Strongyloides stercoralis was 64.3, 59.1, 53.8, 19.4, 8.6, 8.6, and 1.1%, respectively. Only the associations between infection with B. hominis and E. nana with diarrhea were statistically significant. Although B. hominis and E. nana are considered to be nonpathogenic organisms, this study demonstrated that they can be associated with diarrhea in children when they occur at high prevalence and intensity. This survey supports the recent evidence that B. hominis and E. nana infections are associated with deficient sanitation and low hygiene standards and can contribute to diarrhea in children in developing countries. PMID:16249910

  19. A Pilot Study on Single-dose Toxicity Testing of Hominis placenta Pharmacopuncture in Sprague-Dawley Rats

    PubMed Central

    Lee, Yoo-Hwan; Yoon, Hyun-Min; Jang, Kyung-Jeon; Kim, Cheol-Hong

    2015-01-01

    Objectives: This study was performed to analyze the toxicity and to find the lethal dose of the test substance Hominis placenta pharmacopuncture when used as a single-dose in 6 week old, male and female Sprague-Dawley (SD) rats. Methods: All experiments were conducted at Biotoxtech (Chungwon, Korea), an institution authorized to perform non clinical studies, under the regulations of Good Laboratory Practice (GLP). SD rats were chosen for the pilot study. Doses of Hominis placenta pharmacopuncture extracts, 0.125, 0.25 and 0.5 mL, were administered to the experimental group, and 0.5 mL doses of normal saline solution were administered to the control group. This study was conducted under the approval of the Institutional Animal Ethics Committee. Results: No deaths or abnormalities occurred in any of the groups. Also, no significant changes in body weights were observed among the groups, and no significant differences in hematology/biochemistry, necropsy, and histopathology results were noted. Hematologically, some changes in the male rats in two experimental groups were observed, but those changes had no clinical or toxicological meaning because they were not dose dependent. Histopathological tests on the injected parts showed cell infiltration in the male rats in one of the experimental groups; however, that result was due to spontaneous generation and had no toxicological meaning. Therefore, this study showed that Hominis placenta pharmacopuncture had no effect on the injected parts in terms of clinical signs, body weight, hematology, clinical chemistry, and necropsy. Conclusion: As a result of single-dose tests of the test substance Hominis placenta pharmacopuncture in 4 groups of rats, the lethal dose for both males and females exceeded 0.5 mL/animal. Therefore, the above findings suggest that treatment with Hominis placenta pharmacopuncture is relatively safe. Further studies on this subject are needed.

  20. Detection of Tritrichomonas foetus and Pentatrichomonas hominis in intestinal tissue specimens of cats by chromogenic in situ hybridization

    PubMed Central

    Mostegl, Meike M.; Wetscher, Andreas; Richter, Barbara; Nedorost, Nora; Dinhopl, Nora; Weissenböck, Herbert

    2012-01-01

    In this retrospective study 102 cats were analyzed for the presence of trichomonads in intestinal tissue sections using chromogenic in situ hybridization (CISH). Two intestinal trichomonad species are described in cats: Pentatrichomonas hominis and Tritrichomonas foetus. While P. hominis is considered a mere commensal, T. foetus has been found to be the causative agent of feline large-bowel diarrhea. For the detection of both agents within intestinal tissue CISH assays using three different probes were performed. In the first CISH run a probe specific for all relevant members of the order Trichomonadida (OT probe) was used. In a second CISH run all positive samples were further examined on three consecutive tissue sections using the OT probe, a probe specific for the family of Tritrichomonadidae (Tritri probe) and a newly designed probe specifically detecting P. hominis (Penta hom probe). In total, four of the 102 cats were found to be positive with the OT probe. Thereof, one cat gave a positive reaction with the P. hominis probe and three cats were positive with the T. foetus probe. All Trichomonas-positive cats were pure-bred and between 8 and 32 weeks of age. In one cat positive for T. foetus large amounts of parasites were found in the gut lumen and invading the intestinal mucosa. The species of the detected trichomonads were confirmed by polymerase chain reaction and nucleotide sequencing of a part of the 18S ribosomal RNA gene. In this study, the usefulness of CISH to detect intestinal trichomonads within feline tissue samples was shown. Additionally, the specific detection of P. hominis using CISH was established. Generally, it was shown that CISH is well suited for detection and differentiation of trichomonosis in retrospective studies using tissue samples. PMID:21856079

  1. Fatal Outbreak of Mycoplasma capricolum Pneumonia in Endangered Markhors

    PubMed Central

    Thiaucourt, Francois; Amirbekov, Mulojon; Mahmadshoev, Abdurahmon; Manso-Silván, Lucía; Dupuy, Virginie; Vahobov, Dustmurod; Ziyoev, Orom; Michel, Stefan

    2011-01-01

    A pneumonia outbreak reduced the numbers of a wild population of endangered markhors (Capra falconeri) in Tajikistan in 2010. The infection was diagnosed by histologic examination and bacteriologic testing. Mycoplasma capricolum subsp. capricolum was the sole infectious agent detected. Cross-species transmission from domestic goats may have occurred. PMID:22172532

  2. Two Domains within the Mycoplasma hyopneumoniae Cilium Adhesin Bind Heparin

    Microsoft Academic Search

    Cheryl Jenkins; Jody L. Wilton; F. Chris Minion; Linda Falconer; Mark J. Walker; Steven P. Djordjevic

    2006-01-01

    Mycoplasma hyopneumoniae is the causative agent of porcine enzootic pneumonia, a chronic and economically significant respiratory disease that affects swine production worldwide. M. hyopneumoniae adheres to and adversely affects the function of ciliated epithelial cells of the respiratory tract, and the cilium adhesin (Mhp183, P97) is intricately but not exclusively involved in this process. Although binding of pathogenic bacteria to

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

    PubMed Central

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

    2012-01-01

    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. PMID:23543932

  4. A phylogenomic appraisal of the evolutionary relationship of mycoplasmas

    Microsoft Academic Search

    Karla S. C. Yotoko; Sandro L. Bonatto

    2007-01-01

    Several genomes of mycoplasmas have been sequenced and here we tried to retrieve the evolutionary relationships of nine species using a phylogenomic approach. Several methods were used to build phylogenetic trees based on protein sequence information, gene-order, and gene-content. We also utilized datasets composed of individual and concatenated sets of orthologous proteins, as well as with reduced unreliable alignment regions.

  5. Transposon Mutagenesis Identifies Genes Associated with Mycoplasma pneumoniae Gliding Motility

    Microsoft Academic Search

    Benjamin M. Hasselbring; Clinton A. Page; Edward S. Sheppard; Duncan C. Krause

    2006-01-01

    The wall-less prokaryote Mycoplasma pneumoniae, a common cause of chronic respiratory tract infections in humans, is considered to be among the smallest and simplest known cells capable of self-replication, yet it has a complex architecture with a novel cytoskeleton and a differentiated terminal organelle that function in adherence, cell division, and gliding motility. Recent findings have begun to elucidate the

  6. 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

  7. Identification of a ribonuclease H gene in both Mycoplasma genitalium and Mycoplasma pneumoniae by a new method for exhaustive identification of ORFs in the complete genome sequences

    Microsoft Academic Search

    Matthew I Bellgard; Takashi Gojobori

    1999-01-01

    Exhaustive identification of open reading frames in complete genome sequences is a difficult task. It is possible that important genes are missed. In our efforts to reanalyze the intergenic regions of Mycoplasma genitalium and Mycoplasma pneumoniae, we have newly identified a number of new open reading frames (ORFs) in both M. genitalium and M. pneumoniae. The most significant identification was

  8. Protective Immunity against Infection with Mycoplasma haemofelis

    PubMed Central

    Hicks, Chelsea A. E.; Willi, Barbara; Riond, Barbara; Novacco, Marilisa; Meli, Marina L.; Stokes, Christopher R.; Helps, Christopher R.; Hofmann-Lehmann, Regina

    2014-01-01

    Hemoplasmas are potentially zoonotic mycoplasmal pathogens, which are not consistently cleared by antibiotic therapy. Mycoplasma haemofelis is the most pathogenic feline hemoplasma species. The aim of this study was to determine how cats previously infected with M. haemofelis that had recovered reacted when rechallenged with M. haemofelis and to characterize the immune response following de novo M. haemofelis infection and rechallenge. Five specific-pathogen-free (SPF)-derived naive cats (group A) and five cats that had recovered from M. haemofelis infection (group B) were inoculated subcutaneously with M. haemofelis. Blood M. haemofelis loads were measured by quantitative PCR (qPCR), antibody response to heat shock protein 70 (DnaK) by enzyme-linked immunosorbent assay (ELISA), blood lymphocyte cell subtypes by flow cytometry, and cytokine mRNA levels by quantitative reverse transcriptase PCR. Group A cats all became infected with high bacterial loads and seroconverted, while group B cats were protected from reinfection, thus providing the unique opportunity to study the immunological parameters associated with this protective immune response against M. haemofelis. First, a strong humoral response to DnaK was only observed in group A, demonstrating that an antibody response to DnaK is not important for protective immunity. Second, proinflammatory cytokine interleukin-6 (IL-6) mRNA levels appeared to increase rapidly postinoculation in group B, indicating a possible role in protective immunity. Third, an increase in IL-12p35 and -p40 mRNA and decrease in the Th2/Th1 ratio observed in group A suggest that a Th1-type response is important in primary infection. This is the first study to demonstrate protective immunity against M. haemofelis reinfection, and it provides important information for potential future hemoplasma vaccine design. PMID:25410206

  9. Viability of Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri in goat milk samples stored under different conditions.

    PubMed

    Amores, Joaquín; Sánchez, Antonio; Martín, Angel Gómez; Corrales, Juan C; Contreras, Antonio; de la Fe, Christian

    2010-10-26

    Control programs for contagious agalactia (CA) involve monitoring milk samples to detect this disease. This study was designed to establish the effects of the preservatives generally used in dairy laboratories and storage temperature on the viability of Mycoplasma (M.) agalactiae (Ma) and M. mycoides subsp. capri (Mmc) in goat milk samples. In total, 1440 determinations were conducted for each mycoplasma species in milk samples subjected to different storage temperatures (refrigeration at 4°C or freezing at -20°C), preservation strategies (no preservative, NP; azidiol, AZ; or bronopol, BR) and storage times at each temperature (0, 2, 4, 6, 8, 10 and 24h at 4°C and 48h, 1 week, 2 weeks and 4 weeks at -20°C). Our findings reveal the similar viability of Mmc in milk samples stored at 4°C for 24h under the three preservation conditions examined. In contrast, the isolation of Ma in refrigerated milk samples was compromised by the presence of BR, and in smaller measure by the treatments AZ and NP. Freezing milk samples considerably reduced the viability of both mycoplasmas. Given the different sensitivity of the two mycoplasma species to BR, refrigerated milk samples treated with AZ could be used to detect infections caused by both species through culture-based methods. PMID:20413227

  10. High Prevalence of Mycoplasma Infections in Symptomatic (Chronic Fatigue Syndrome) Family Members of Mycoplasma-Positive Gulf War Illness Patients

    Microsoft Academic Search

    Garth L. Nicolson; Marwan Y. Nasralla; Nancy L. Nicolson; Joerg Haier

    2003-01-01

    SUMMARY. Immediate family members of veterans diagnosed with Gulf War Illnesses often complain of fatiguing illnesses, and upon analysis they report similar signs and symptoms as their veteran family members. Since a relatively common finding in Gulf War Illness patients is a bacterial infection due to Mycoplasma species, we examined military families (149 patients: 42 veterans, 40 spouses, 32 other

  11. [PCR-based genotype classification of Blastocystis hominis isolates from college students of Guangxi].

    PubMed

    Zhan, Ting-Zheng; Liu, Teng; Shi, Huan-Huan; He, Shan-Shan; Yan, Hui; Liu, Deng-Yu

    2014-06-01

    Fifty-three Blastocystis hominis isolates were separated from the fecal specimens of carriers in college students from Guangxi and cultivated in vitro, and the genetic DNA was extracted. All the isolates were genotyped by PCR using seven pairs of known sequence-tagged site (STS) primers. The results showed there were five subtypes in the 53 isolates. Subtype 3 was the most popular one (32.1%, 17/53), followed by subtype 7 (9.4%, 5/53). Subtypes 1 (7.6%, 4/53), 4 (7.6%, 4/53), and 6 (1.9%, 1/53) were detected, while subtypes 2 and 5 were not detected. The genotypes of the other 22 isolates were unknown which were negative to all the STS primers. PMID:25223057

  12. 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:23145790

  13. Cloning, Expression, and Immunological Characterization of the P30 Protein of Mycoplasma pneumoniae?

    PubMed Central

    Varshney, Avanish Kumar; Chaudhry, Rama; Kabra, Sushil Kumar; Malhotra, Pawan

    2008-01-01

    Mycoplasma pneumoniae, a self-replicating cell wall-deficient prokaryote, has a differentiated terminal organelle that is essential for cytadherence and gliding motility. P30, an important protein associated with the terminal organelle, is required for the cytadherence and virulence of M. pneumoniae. P30 is a transmembrane protein with an intracytoplasmic N terminus and an exposed C terminus. In the present study, we amplified and sequenced the full-length p30 gene of Mycoplasma pneumoniae directly from 18 Indian asthmatic patients. Sequence diversity was observed in the p30 genes from 16 clinical samples when the sequences were compared with the sequence of strain M-129. We also successfully expressed a fragment of the p30 gene (P30B) that includes the complete C-terminal proline-rich amino acid sequences in different Escherichia coli expression systems. The maltose binding protein (MBP)-P30B fusion protein was recognized by M. pneumoniae-infected patient sera in immunoblots, and the protein was immunogenic in mice. We further analyzed the reactivity of the MBP-P30B fusion protein with patient sera in an enzyme-linked immunosorbent assay (ELISA) and compared it with the reactivity obtained with a commercial kit (the Serion ELISA Classic kit). The sensitivity and the specificity of the in-house ELISA were 78.57% and 89.47%, respectively. This study suggests that the P30 protein can be used as an antigen along with other adhesin proteins for the immunodiagnosis of M. pneumoniae infection. PMID:18032594

  14. Cloning, expression, and immunological characterization of the P30 protein of Mycoplasma pneumoniae.

    PubMed

    Varshney, Avanish Kumar; Chaudhry, Rama; Kabra, Sushil Kumar; Malhotra, Pawan

    2008-02-01

    Mycoplasma pneumoniae, a self-replicating cell wall-deficient prokaryote, has a differentiated terminal organelle that is essential for cytadherence and gliding motility. P30, an important protein associated with the terminal organelle, is required for the cytadherence and virulence of M. pneumoniae. P30 is a transmembrane protein with an intracytoplasmic N terminus and an exposed C terminus. In the present study, we amplified and sequenced the full-length p30 gene of Mycoplasma pneumoniae directly from 18 Indian asthmatic patients. Sequence diversity was observed in the p30 genes from 16 clinical samples when the sequences were compared with the sequence of strain M-129. We also successfully expressed a fragment of the p30 gene (P30B) that includes the complete C-terminal proline-rich amino acid sequences in different Escherichia coli expression systems. The maltose binding protein (MBP)-P30B fusion protein was recognized by M. pneumoniae-infected patient sera in immunoblots, and the protein was immunogenic in mice. We further analyzed the reactivity of the MBP-P30B fusion protein with patient sera in an enzyme-linked immunosorbent assay (ELISA) and compared it with the reactivity obtained with a commercial kit (the Serion ELISA Classic kit). The sensitivity and the specificity of the in-house ELISA were 78.57% and 89.47%, respectively. This study suggests that the P30 protein can be used as an antigen along with other adhesin proteins for the immunodiagnosis of M. pneumoniae infection. PMID:18032594

  15. Novel strategy for typing Mycoplasma pneumoniae isolates by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry coupled with ClinProTools.

    PubMed

    Xiao, Di; Zhao, Fei; Zhang, Huifang; Meng, Fanliang; Zhang, Jianzhong

    2014-08-01

    The typing of Mycoplasma pneumoniae mainly relies on the detection of nucleic acid, which is limited by the use of a single gene target, complex operation procedures, and a lengthy assay time. Here, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) coupled to ClinProTools was used to discover MALDI-TOF MS biomarker peaks and to generate a classification model based on a genetic algorithm (GA) to differentiate between type 1 and type 2 M. pneumoniae isolates. Twenty-five M. pneumoniae strains were used to construct an analysis model, and 43 Mycoplasma strains were used for validation. For the GA typing model, the cross-validation values, which reflect the ability of the model to handle variability among the test spectra and the recognition capability value, which reflects the model's ability to correctly identify its component spectra, were all 100%. This model contained 7 biomarker peaks (m/z 3,318.8, 3,215.0, 5,091.8, 5,766.8, 6,337.1, 6,431.1, and 6,979.9) used to correctly identify 31 type 1 and 7 type 2 M. pneumoniae isolates from 43 Mycoplasma strains with a sensitivity and specificity of 100%. The strain distribution map and principle component analysis based on the GA classification model also clearly showed that the type 1 and type 2 M. pneumoniae isolates can be divided into two categories based on their peptide mass fingerprints. With the obvious advantages of being rapid, highly accurate, and highly sensitive and having a low cost and high throughput, MALDI-TOF MS ClinProTools is a powerful and reliable tool for M. pneumoniae typing. PMID:24920781

  16. A specific PCR for the identification of Mycoplasma capricolum subsp. capripneumoniae, the causative agent of contagious caprine pleuropneumonia (CCPP).

    PubMed

    Woubit, S; Lorenzon, S; Peyraud, A; Manso-Silván, L; Thiaucourt, F

    2004-11-30

    Contagious caprine pleuropneumonia is a severe infectious disease of goats in Africa and the Middle East. It is caused by a fastidious mycoplasma, Mycoplasma capricolum subsp. capripneumoniae, a member of the "M. mycoides cluster". Members of this cluster share genomic and antigenic features, which result in common biochemical and serological properties, complicating species identification. Two species of this cluster, M. mycoides subsp. capri and M. mycoides subsp. mycoides large colony biotype, are very often isolated from clinical cases resembling contagious caprine pleuropneumonia. Furthermore, in the laboratory, M. capricolum subsp. capripneumoniae can be easily confused with the closely related capricolum subspecies. Considering these constraints and the scarcity of available methods for identification, a specific polymerase chain reaction was developed. A DNA fragment of 7109 bp containing genes coding for the arginine deiminase pathway (ADI) was chosen as target sequence for the selection of a specific primer pair. The full ADI operon from M. capricolum subsp. capripneumoniae strain GL100 was sequenced. Polymorphism within this locus was analyzed by comparison with the sequence from the closely related IPX strain (M. capricolum subsp. capricolum). It varied from 0.6% to 3.5%. The highest divergence was found in a region coding for arcD. Therefore, this gene was chosen as target for the specific amplification of a 316 bp-long DNA fragment. The specificity of this PCR was validated on 14 M. capricolum subsp. capripneumoniae strains and 27 heterologous strains belonging to the "M. mycoides cluster" and M. putrefaciens. This new PCR will be a valuable tool for the surveillance of contagious caprine pleuropneumonia. PMID:15530747

  17. 9 CFR 147.16 - Procedure for the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment).

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...when: (1) Test birds have serum plate antibodies for the mycoplasma for which the donor birds were tested, regardless of HI test results, and control birds stay serologically negative; or (2) Mycoplasma organisms are isolated from the test...

  18. 9 CFR 147.16 - Procedure for the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment).

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...when: (1) Test birds have serum plate antibodies for the mycoplasma for which the donor birds were tested, regardless of HI test results, and control birds stay serologically negative; or (2) Mycoplasma organisms are isolated from the test...

  19. 9 CFR 147.16 - Procedure for the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment).

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...when: (1) Test birds have serum plate antibodies for the mycoplasma for which the donor birds were tested, regardless of HI test results, and control birds stay serologically negative; or (2) Mycoplasma organisms are isolated from the test...

  20. Secretory immune responses to Mycoplasma pulmonis.

    PubMed Central

    Steffen, M J; Ebersole, J L

    1992-01-01

    Formalinized Mycoplasma pulmonis, along with aluminum hydroxide as an adjuvant, was used to subcutaneously immunize rats in the vicinity of the salivary gland to examine the characteristics of the secretory immune response to this pathogen. The induction of specific antibody to this microorganism was detected in serum and the exocrine fluids, namely, saliva and lung lavage fluid. Both immunoglobulin G (IgG) and IgA isotype antibodies were detected in each of these fluids after primary and secondary local immunizations. Serum responses from immunized animals were significantly greater than in the control group, but a dose response was not observed in either IgG or IgA antibody at the dosages selected for immunization. Salivary IgG antibody responses peaked early after both the primary and secondary immunizations, exhibiting a clear dose response. Salivary IgA in immunized groups was significantly greater than that in the control group but displayed little dose-dependent kinetics, and, at the termination of the experiment, this response had not yet peaked. Lung lavage IgG and IgA were minimal after the primary immunization when the antibody was normalized to total protein but displayed dose-dependent kinetics after a secondary challenge. IgG peaked immediately after a secondary challenge, while IgA peak responses were observed only after 20 days. A positive correlation was noted between the serum, saliva, and lung lavage fluid IgGs after both primary and secondary immunizations and only after a secondary challenge for IgA. In this study we were able to elicit a secretory immune response, consisting of both IgG and IgA, which exhibited a dose-dependent characteristic in lung lavage fluid to this immunogen. Additionally, a positive correlation of antibody levels between saliva and lung lavage fluid suggests that saliva could be used as an indicator for monitoring specific antibody to M. pulmonis in lung lavage secretions without requiring invasive, deleterious procedures. PMID:1730465

  1. 9 CFR 147.16 - Procedure for the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment).

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ...evaluation of mycoplasma reactors by in vivo bio-assay (enrichment). 147.16 Section...evaluation of mycoplasma reactors by in vivo bio-assay (enrichment). This procedure...Bigland, C. H. and A. J. DaMassa, “A Bio-Assay for Mycoplasma...

  2. Phase-locked mutants of Mycoplasma agalactiae: defining the molecular switch of high-frequency Vpma antigenic variation

    PubMed Central

    Chopra-Dewasthaly, Rohini; Citti, Christine; Glew, Michelle D; Zimmermann, Martina; Rosengarten, Renate; Jechlinger, Wolfgang

    2008-01-01

    Mycoplasma agalactiae, an important pathogen of small ruminants, exhibits antigenic diversity by switching the expression of multiple surface lipoproteins called Vpmas (Variable proteins of M. agalactiae). Although phase variation has been shown to play important roles in many host–pathogen interactions, the biological significance and the mechanism of Vpma oscillations remain largely unclear. Here, we demonstrate that all six Vpma proteins are expressed in the type strain PG2 and all undergo phase variation at an unusually high frequency. Furthermore, targeted gene disruption of the xer1 gene encoding a putative site-specific recombinase adjacent to the vpma locus was accomplished via homologous recombination using a replicon-based vector. Inactivation of xer1 abolished further Vpma switching and the ‘phase-locked’ mutants (PLMs) continued to steadily express only a single Vpma product. Complementation of the wild-type xer1 gene in PLMs restored Vpma phase variation thereby proving that Xer1 is essential for vpma inversions. The study is not only instrumental in enhancing our ability to understand the role of Vpmas in M. agalactiae infections but also provides useful molecular approaches to study potential disease factors in other ‘difficult-to-manipulate’ mycoplasmas. PMID:18248580

  3. Detection of Mycoplasma pneumoniae in Simulated and True Clinical Throat Swab Specimens by Nanorod Array-Surface-Enhanced Raman Spectroscopy

    PubMed Central

    Hennigan, Suzanne L.; Driskell, Jeremy D.; Dluhy, Richard A.; Zhao, Yiping; Tripp, Ralph A.; Waites, Ken B.; Krause, Duncan C.

    2010-01-01

    The prokaryote Mycoplasma pneumoniae is a major cause of respiratory disease in humans, accounting for 20% of all community-acquired pneumonia and the leading cause of pneumonia in older children and young adults. The limitations of existing options for mycoplasma diagnosis highlight a critical need for a new detection platform with high sensitivity, specificity, and expediency. Here we evaluated silver nanorod arrays (NA) as a biosensing platform for detection and differentiation of M. pneumoniae in culture and in spiked and true clinical throat swab samples by surface-enhanced Raman spectroscopy (SERS). Three M. pneumoniae strains were reproducibly differentiated by NA-SERS with 95%–100% specificity and 94–100% sensitivity, and with a lower detection limit exceeding standard PCR. Analysis of throat swab samples spiked with M. pneumoniae yielded detection in a complex, clinically relevant background with >90% accuracy and high sensitivity. In addition, NA-SERS correctly classified with >97% accuracy, ten true clinical throat swab samples previously established by real-time PCR and culture to be positive or negative for M. pneumoniae. Our findings suggest that the unique biochemical specificity of Raman spectroscopy, combined with reproducible spectral enhancement by silver NA, holds great promise as a superior platform for rapid and sensitive detection and identification of M. pneumoniae, with potential for point-of-care application. PMID:21049032

  4. A unique case of facial burn superinfected with Dermatobia Hominis larvae resulting in a bilateral enucleation of the eyes.

    PubMed

    Pinos, Victor Hugo; Ortiz-Prado, Esteban; Bermeo, Carlos; León, Juan; Armijos, Luciana; Almeida, Estibaliz

    2014-10-01

    We present a case of a female Ecuadorian patient who presented a deep facial burn injury complicated with a severe infestation of Dermatobia Hominis larvae. The burn injury was complicated by severe myiasis attributable to the poor management of the wound received at home, using tropical plants, which caused a secondary infection and severe necrosis of the tissue involving the forehead, cheeks, chin, scalp, nose, mouth and the eyes resulting in a bilateral enucleation and long inpatient hospital care. PMID:24728977

  5. Relationships Between Mycoplasma pneumoniae and Human Respiratory Epithelium

    PubMed Central

    Collier, Albert M.; Clyde, Wallace A.

    1971-01-01

    The interaction was studied between Mycoplasma pneumoniae and its natural host cell, the human respiratory epithelium. Organized, ciliated cells provided by fetal trachea in organ culture enabled examination of the host-parasite relationship by light, immunofluorescence, and electron microscopy. Impairment of cellular function was reflected by disorganization and loss of ciliary motion; this was associated with a sequence of cytopathological changes denoting progressive cell injury. The organisms were found concentrated on the luminal surface of ciliated epithelium and cells lining the submucosal glands. A differentiated portion of the Mycoplasma, consisting of an extension of the unit membrane containing an electron-dense core surrounded by a lucent space, served as the means of attachment to host cells. The findings suggest that the pathogenicity of M. pneumoniae depends upon intimate extracellular infection with production of functional and structural changes initiated by host cell membrane injury. Images PMID:16558038

  6. The Mycoplasma conjunctivae genome sequencing, annotation and analysis

    PubMed Central

    Calderon-Copete, Sandra P; Wigger, George; Wunderlin, Christof; Schmidheini, Tobias; Frey, Joachim; Quail, Michael A; Falquet, Laurent

    2009-01-01

    Background The mollicute Mycoplasma conjunctivae is the etiological agent leading to infectious keratoconjunctivitis (IKC) in domestic sheep and wild caprinae. Although this pathogen is relatively benign for domestic animals treated by antibiotics, it can lead wild animals to blindness and death. This is a major cause of death in the protected species in the Alps (e.g., Capra ibex, Rupicapra rupicapra). Methods The genome was sequenced using a combined technique of GS-FLX (454) and Sanger sequencing, and annotated by an automatic pipeline that we designed using several tools interconnected via PERL scripts. The resulting annotations are stored in a MySQL database. Results The annotated sequence is deposited in the EMBL database (FM864216) and uploaded into the mollicutes database MolliGen allowing for comparative genomics. Conclusion We show that our automatic pipeline allows for annotating a complete mycoplasma genome and present several examples of analysis in search for biological targets (e.g., pathogenic proteins). PMID:19534756

  7. Multilocus sequence typing of Mycoplasma bovis reveals host-specific genotypes in cattle versus bison.

    PubMed

    Register, Karen B; Thole, Luke; Rosenbush, Ricardo F; Minion, F Chris

    2015-01-30

    Mycoplasma bovis is a primary agent of mastitis, pneumonia and arthritis in cattle and the bacterium most frequently isolated from the polymicrobial syndrome known as bovine respiratory disease complex. Recently, M. bovis has emerged as a significant health problem in bison, causing necrotic pharyngitis, pneumonia, dystocia and abortion. Whether isolates from cattle and bison comprise genetically distinct populations is unknown. This study describes the development of a highly discriminatory multilocus sequencing typing (MLST) method for M. bovis and its use to investigate the population structure of the bacterium. Genome sequences from six M. bovis isolates were used for selection of gene targets. Seven of 44 housekeeping genes initially evaluated were selected as targets on the basis of sequence variability and distribution within the genome. For each gene target sequence, four to seven alleles could be distinguished that collectively define 32 sequence types (STs) from a collection of 94 cattle isolates and 42 bison isolates. A phylogeny based on concatenated target gene sequences of each isolate revealed that bison isolates are genetically distinct from strains that infect cattle, suggesting recent disease outbreaks in bison may be due to the emergence of unique genetic variants. No correlation was found between ST and disease presentation or geographic origin. MLST data reported here were used to populate a newly created and publicly available, curated database to which researchers can contribute. The MLST scheme and database provide novel tools for exploring the population structure of M. bovis and tracking the evolution and spread of strains. PMID:25433454

  8. The identification of mycoplasma species by gas chromatography 

    E-print Network

    Shult, Milton Donald

    1969-01-01

    method to identify myco- plasma at the species level because of their minimal bio- chemical activity and their pleomorphism . Recent reports have indicated that the use of gas-liquid chromatography of cellular components is feasible...THE IDENTIFICATION OF MYCOPLASMA SPECIES BY GAS CHROMATOGRAPHY A Thesis MILTON DONALD SHULT& JR. Submitted to the Graduate College of Texas AFM University in partial fulfillment of the requirement for the degree of MASTER OF SCIENCE May 1969...

  9. 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 ?(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 antigenic switch constitutes a finely tuned mechanism that may be involved in virulence. PMID:23869216

  10. Mycoplasma pneumoniae associated opsoclonus–myoclonus syndrome in three cases

    Microsoft Academic Search

    Benedikt Maria Huber; Susi Strozzi; Maja Steinlin; Christoph Aebi; Simon Fluri

    2010-01-01

    Opsoclonus–myoclonus syndrome (OMS) is a rare acquired movement disorder occurring in all age groups, predominantly in infants.\\u000a Although the exact pathogenesis is still undefined, there is strong evidence for a paraneoplastic or parainfectious immune\\u000a process resulting in central nervous system dysfunction. Mycoplasma pneumoniae has been implicated in a number of immune-mediated neurologic diseases [28]. However, the association of M. pneumoniae

  11. Anti-galactocerebroside testing in Mycoplasma pneumoniae-associated encephalitis

    Microsoft Academic Search

    Laura J. Christie; Somayeh Honarmand; Shigeo Yagi; Sara Ruiz; Carol A. Glaser

    2007-01-01

    Mycoplasma pneumoniae (Mp) is the most frequently identified pathogen in the California Encephalitis Project, but the role and mechanism of Mp is unclear. Since auto-antibodies to anti-galactocerebroside (anti-GalC) have been reported in patients with evidence of acute Mp infection of the central nervous system (CNS), serum and cerebrospinal fluid (CSF) samples from 26 patients with evidence of Mp were tested

  12. Mycoplasma pneumoniae Pneumonia Associated With Methemoglobinemia and Anemia: An Overlooked Association?

    PubMed Central

    Khoury, Tawfik; Abu Rmeileh, Ayman; Kornspan, Jonathan David; Abel, Roy; Mizrahi, Meir; Nir-Paz, Ran

    2015-01-01

    We report a case of acute methemoglobinemia and anemia in a patient with Mycoplasma pneumoniae pneumonia. We suggest that M. pneumoniae secretes a putative protein that can induce methemoglobin in red blood cells. Thus, Mycoplasma pneumoniae may induce methemoglobinemia in patients who have low oxygen saturation and anemia.

  13. Diagnosis of Mycoplasma pneumoniae Infection in Autopsy and Open-Lung Biopsy Tissues by Nested PCR

    Microsoft Academic Search

    DEBORAH F. TALKINGTON; W. LANIER THACKER; DAVID W. KELLER; JØRGEN S. JENSEN

    1998-01-01

    A nested PCR specific for the Mycoplasma pneumoniae P1 gene was used to diagnose mycoplasma infection in two cohort patients with severe pneumonia within 24 h of tissue receipt. A postmortem diagnosis of M. pneumoniae infection was obtained for the first patient, who died without the collection of appropriate paired samples for serodiagnosis. An open-lung biopsy obtained from the second

  14. Draft Genome Sequence of the First Human Isolate of the Ruminant Pathogen Mycoplasma capricolum subsp. capricolum.

    PubMed

    Seersholm, Frederik Valeur; Fischer, Anne; Heller, Martin; Jores, Joerg; Sachse, Konrad; Mourier, Tobias; Hansen, Anders Johannes

    2015-01-01

    Mycoplasma capricolum subsp. capricolum is a well-known pathogen of small ruminants. A recent human case of septicemia involving this agent raised the question of its potential pathogenicity to humans. We present the first draft genome sequence of a human Mycoplasma capricolum subsp. capricolum isolate. PMID:26089408

  15. Draft Genome Sequence of the First Human Isolate of the Ruminant Pathogen Mycoplasma capricolum subsp. capricolum

    PubMed Central

    Fischer, Anne; Heller, Martin; Jores, Joerg; Sachse, Konrad; Mourier, Tobias; Hansen, Anders Johannes

    2015-01-01

    Mycoplasma capricolum subsp. capricolum is a well-known pathogen of small ruminants. A recent human case of septicemia involving this agent raised the question of its potential pathogenicity to humans. We present the first draft genome sequence of a human Mycoplasma capricolum subsp. capricolum isolate. PMID:26089408

  16. Gaussia luciferase-based mycoplasma detection assay in mammalian cell culture.

    PubMed

    Degeling, M Hannah; Bovenberg, M Sarah S; Tannous, Marie; Tannous, Bakhos A

    2014-01-01

    Mycoplasma contamination in mammalian cell culture is a common problem with serious consequences on experimental data, and yet many laboratories fail to perform regular testing. In this chapter, we describe a simple and sensitive mycoplasma detection assay based on the bioluminescent properties of the Gaussia luciferase reporter. PMID:24166367

  17. Dual infection with Mycoplasma synoviae and a tenosynovitis?inducing reovirus in chickens

    Microsoft Academic Search

    Janet M. Bradbury; Antonella Garuti

    1978-01-01

    One group of embryonated chicken eggs was inoculated with sterile myco?plasma broth and another with a broth culture of Mycoplasma synoviae. Each group was subdivided into two and the chicks were hatched in four isolation pens. At 1 day of age one uninfected and one M. synoviae infected group were inoculated with a reovirus known to cause tenosynovitis. Birds were

  18. Gene and cytokine profile analysis of macrolide-resistant Mycoplasma pneumoniae infection in Fukuoka, Japan

    PubMed Central

    2013-01-01

    Background Recent epidemiologic data suggest that the prevalence of macrolide resistant Mycoplasma pneumoniae (MR-M. pneumoniae) is increasing rapidly worldwide. This study assessed the present status of M. pneumoniae infection in Japan and clinical end-points to distinguish children with MR-M. pneumoniae. Methods During an outbreak of M. pneumoniae infections in Fukuoka, Japan in 2010–11, a total of 105 children with clinically suspected M. pneumoniae infection were enrolled. M. pneumoniae was analyzed for macrolide resistance in domain V of the 23S rRNA gene. Sixty -five patients with PCR positive for M. pneumoniae were analyzed with regard to clinical symptoms, efficacy of several antimicrobial agents and several laboratory data. Results Causative pathogens were detected in 81.0% (85 of 105) and M. pneumoniae was identified 61.9% (65 of 105). The resistance rate of M. pneumoniae was 89.2% (58 of 65) in this general pediatric outpatient setting. Patients infected with MR-M. pneumoniae showed longer times to resolution of fever and required frequent changes of the initially prescribed macrolide to another antimicrobial agent. We observed three different genotypes of M. pneumoniae including the rarely reported A2063T mutation (A2063G: 31 strains, A2063T: 27 strains, no mutation: 7 strains). Drug susceptibility testing showed different antimicrobial susceptibility profiles for each genotype. Serum IFN-gamma, IL-6 and IP-10 levels were higher in patients with MR-genotypes than in those infected with no-mutation strains (p?

  19. 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). PMID:18957640

  20. Molecular identification of Mycoplasma cynos from laboratory beagle dogs with respiratory disease

    PubMed Central

    Hong, Sunhwa

    2012-01-01

    In this study, we examined a colony of 20 beagle dogs in a laboratory animal facility. Mycoplasma was detected by consensus PCR assay in 1 dog with respiratory and constitutional symptoms. None of the other dogs were affected. The dog was euthanized and necropsied. In postmortem examinations, gray or plum-colored gross lesions were found on the lung, most commonly in the apical and cardiac lobes. Some lesions showed clear demarcation and consolidation. Microscopic examination showed peribronchiolar lymphoid hyperplasia and interstitial thickening, lesions pathognomonic for mycoplasma pneumonia. To identify canine Mycoplasma species, we used species-specific PCR reactions for M. arginini, M. canis, M. cynos, M. edwardii, M. felis, M. gateae, M. maculosum, M. molare, M. opalescens, M. spumans, Mycoplasma sp. HRC 689, and M. collis. As the result, we identified Mycoplasma cynos by amplification of DNA extracted from lung tissue of the laboratory beagle dog with respiratory disease. PMID:22474476

  1. Heterogeneity and Compartmentalization of Pneumocystis carinii f. sp. hominis Genotypes in Autopsy Lungs

    PubMed Central

    Helweg-Larsen, Jannik; Lundgren, Bettina; Lundgren, Jens D.

    2001-01-01

    The extent and importance of genotype heterogeneity of Pneumocystis carinii f. sp. hominis within lungs have not previously been investigated. Two hundred forty PCR clones obtained from respiratory specimens and lung segments from three patients with fatal P. carinii pneumonia were investigated to detect genetic diversity in the internal transcribed spacer (ITS) region of the nuclear rRNA operon, the mitochondrial large-subunit (mtLSU) rRNA gene, and the dihydropteroate synthase-encoding gene. For two of the three examined patients, a mixture of different mtLSU rRNA and ITS genotypes was observed. Not all genotypes present in the lungs at autopsy were detected in the diagnostic respiratory samples. Compartmentalization of specific ITS and mtLSU rRNA sequence types was observed in different lung segments. In conclusion, the interpretation of genotype data and in particular ITS sequence types in the assessment of epidemiological questions should be cautious since genotyping done on respiratory samples cannot a priori be assumed to represent all genotypes present within the lung. PMID:11574620

  2. [Reclassification of the four China isolated strains of the pathogen for contagious caprine pleuropneumonia].

    PubMed

    Li, Yuan; Zhang, Jian-Hua; Hu, Shou-Ping; Wang, Liang; Xin, Jiu-Qing

    2007-10-01

    Contagious caprine pleuropneumonia (CCPP) is caused by Mycoplasma capricolum subsp. capripneumoniae (Mccp). The aims of this study were to identify 4 Chinese isolated strains employing molecular methods and to determine the appropriate subspecies classification of these strains. Three genome fragments (A, B and C) from each strain were amplified and then transformed into plasmids. The inserted fragments were sequenced and analyzed by comparison with six members of the Mycoplasma mycoides cluster. Cleavage of the PCR products of the 4 strains with PstI yielded three fragments 548, 420 and 128bp in length, just like strain F38. The other M. mycoides cluster members had only 2 fragments of 428 and 128bp. Homology analysis of fragment B indicated that the 4 strains exhibited 99.5% homology with Mccp reference strain F38, 98.9% with M. capricolum subsp. Capricolum (Mcc) strain California Kid, and only 95.4% with Mmc strain ZZ. In fragment C, the 4 strains had 67.4% - 67.6% homology with Mmc PG3, 95.1% -98.6% with Mcc strains 8601-50 and California Kid, 99.6% - 99.8% with Mccp strains 97097ET, Gabes and F38. The analysis revealed that 4 pathogeny strains, 87001, 87002, 367, 1653, isolated from China are more closely related to Mccp than to Mcc. Therefore the pathogeny of CCPP in China should be reclassified as Mccp. PMID:18062246

  3. Concordance between genetic relatedness and phenotypic similarities of Trichomonas vaginalis strains

    PubMed Central

    Hampl, Vladimír; Va?á?ová, Št?pánka; Kulda, Jaroslav; Flegr, Jaroslav

    2001-01-01

    Background Despite the medical importance of trichomoniasis, little is known about the genetic relatedness of Trichomonas vaginalis strains with similar biological characteristics. Furthermore, the distribution of endobionts such as mycoplasmas or Trichomonas vaginalis virus (TVV) in the T. vaginalis metapopulation is poorly characterised. Results We assayed the relationship between 20 strains of T. vaginalis from 8 countries using the Random Amplified Polymorphic DNA (RAPD) analysis with 27 random primers. The genealogical tree was constructed and its bootstrap values were computed using the program FreeTree. Using the permutation tail probability tests we found that the topology of the tree reflected both the pattern of resistance to metronidazole (the major anti-trichomonal drug) (p < 0.01) and the pattern of infection of strains by mycoplasmas (p < 0.05). However, the tree did not reflect pattern of virulence, geographic origin or infection by TVV. Despite low bootstrap support for many branches, the significant clustering of strains with similar drug susceptibility suggests that the tree approaches the true genealogy of strains. The clustering of mycoplasma positive strains may be an experimental artifact, caused by shared RAPD characters which are dependent on the presence of mycoplasma DNA. Conclusions Our results confirmed both the suitability of the RAPD technique for genealogical studies in T. vaginalis and previous conclusions on the relatedness of metronidazol resistant strains. However, our studies indicate that testing analysed strains for the presence of endobionts and assessment of the robustness of tree topologies by bootstrap analysis seem to be obligatory steps in such analyses. PMID:11734059

  4. 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. PMID:25088052

  5. Development and validation of an attenuated Mycoplasma hyopneumoniae aerosol vaccine.

    PubMed

    Feng, Zhi-Xin; Wei, Yan-Na; Li, Gui-Lan; Lu, Xiao-Ming; Wan, Xiu-Feng; Pharr, G Todd; Wang, Zhan-Wei; Kong, Meng; Gan, Yuan; Bai, Fang-Fang; Liu, Mao-Jun; Xiong, Qi-Yan; Wu, Xu-Su; Shao, Guo-Qing

    2013-12-27

    Mycoplasma hyopneumoniae (M. hyopneumoniae) causes a chronic respiratory disease with high morbidity and low mortality in swine, and has been presented as a major cause of growth retardation in the swine industry. Aerosol vaccination presents a needle free, high throughput, and efficient platform for vaccine delivery, and has been widely applied in poultry vaccination. However, aerosol vaccines have rarely been used in swine vaccination primarily because the long and curving respiratory track of swine presents a barrier for vaccine particle delivery. To develop an effective M. hyopneumoniae aerosol vaccine, three major barriers need to be overcome: to optimize particle size for aerosol delivery, to maintain the viability of mycoplasma cells in the vaccine, and to optimize the environmental conditions for vaccine delivery. In this study, an aerosol mycoplasma vaccine was successfully developed based on a conventional live attenuated M. hyopneumoniae vaccine. Specifically, the Pari LCD nebulizer was used to produce an aerosol vaccine particle size less than 5 ?m; and a buffer with 5% glycerol was developed and optimized to prevent inactivation of M. hyopneumoniae caused by aerosolization and evaporation. Before nebulization, the room temperature and relative humidity were control to 20-25 °C and 70-75%, respectively, which helped maintain the viability of aerosol vaccine. Animal experiments demonstrated that this newly developed aerosol vaccine was effectively delivered to swine low respiratory track, being confirmed by nested-PCR, in situ hybridization and scanning electron microscope. Moreover, M. hyopneumoniae specific sIgA secretion was detected in the nasal swab samples at 14 days post-immunization. To our knowledge, this is the first report on a live M. hyopneumoniae aerosol vaccine. PMID:24035264

  6. Mycoplasma agalactiae, an Etiological Agent of Contagious Agalactia in Small Ruminants: A Review.

    PubMed

    Kumar, Amit; Rahal, Anu; Chakraborty, Sandip; Verma, Amit Kumar; Dhama, Kuldeep

    2014-01-01

    Mycoplasma agalactiae is one of the causal agents of classical contagious agalactia (CA), a serious, economically important but neglected enzootic disease of small ruminants. It occurs in many parts of the world and most notably in the Mediterranean Basin. Following the infection common complications are septicaemia, mastitis, arthritis, pleurisy, pneumonia, and keratoconjunctivitis. Primary or tentative diagnosis of the organism is based upon clinical signs. Various serological tests, namely, growth precipitation, immunofluorescence, complement fixation test, haemagglutination inhibition, agglutination, immunodiffusion, enzyme immunoassays, immunoelectrophoresis, blotting techniques, and others, are available. Molecular tools seem to be much more sensitive, specific, and faster and help to differentiate various strains. The real-time PCR, multiplex PCR, quantitative PCR, PCR-RFLP, MLST, and gene probes, complementary to segments of chromosomal DNA or 16S ribosomal RNA (rRNA), have strengthened the diagnosis of M. agalactiae. Both live attenuated and adjuvant (alum precipitated or saponified) inactivated vaccines are available with greater use of inactivated ones due to lack of side effects. The present review discusses the etiology, epidemiology, pathogenesis, and clinical signs of contagious agalactia in small ruminants along with trends and advances in its diagnosis, treatment, vaccination, prevention, and control strategies that will help in countering this disease. PMID:25097796

  7. 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

  8. Eggshell apex abnormalities associated with Mycoplasma synoviae infection in layers.

    PubMed

    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; Mo, In-Pil

    2014-12-01

    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

  9. Meningoencephalitis in commercial meat turkeys associated with Mycoplasma gallisepticum.

    PubMed

    Chin, R P; Daft, B M; Meteyer, C U; Yamamoto, R

    1991-01-01

    Mycoplasma gallisepticum (MG) infection was diagnosed in three different flocks of 12-to-16-week-old commercial meat turkeys displaying torticollis and/or opisthotonos. MG was isolated from the brain, air sacs, trachea, and sinus of one bird with neurological signs. Histological examination of brains in all three cases revealed moderate-to-severe encephalitis with lymphoplasmacytic cuffing of vessels, fibrinoid vasculitis, focal parenchymal necrosis, and meningitis. Birds with neurological signs were seropositive for MG by the serum-plate agglutination and hemagglutination-inhibition tests. The encephalitic form of MG has been described previously but is rarely mentioned in the current literature. PMID:1786029

  10. ?-D-Glucoside utilization by Mycoplasma mycoides subsp. mycoides SC: possible involvement in the control of cytotoxicity towards bovine lung cells

    PubMed Central

    Vilei, Edy M; Correia, Ivone; Ferronha, M Helena; Bischof, Daniela F; Frey, Joachim

    2007-01-01

    Background Contagious bovine pleuropneumonia (CBPP) caused by Mycoplasma mycoides subsp. mycoides small-colony type (SC) is among the most serious threats for livestock producers in Africa. Glycerol metabolism-associated H2O2 production seems to play a crucial role in virulence of this mycoplasma. A wide number of attenuated strains of M. mycoides subsp. mycoides SC are currently used in Africa as live vaccines. Glycerol metabolism is not affected in these vaccine strains and therefore it does not seem to be the determinant of their attenuation. A non-synonymous single nucleotide polymorphism (SNP) in the bgl gene coding for the 6-phospho-?-glucosidase (Bgl) has been described recently. The SNP differentiates virulent African strains isolated from outbreaks with severe CBPP, which express the Bgl isoform Val204, from strains to be considered less virulent isolated from CBPP outbreaks with low mortality and vaccine strains, which express the Bgl isoform Ala204. Results Strains of M. mycoides subsp. mycoides SC considered virulent and possessing the Bgl isoform Val204, but not strains with the Bgl isoform Ala204, do trigger elevated levels of damage to embryonic bovine lung (EBL) cells upon incubation with the disaccharides (i.e., ?-D-glucosides) sucrose and lactose. However, strains expressing the Bgl isoform Val204 show a lower hydrolysing activity on the chromogenic substrate p-nitrophenyl-?-D-glucopyranoside (pNPbG) when compared to strains that possess the Bgl isoform Ala204. Defective activity of Bgl in M. mycoides subsp. mycoides SC does not lead to H2O2 production. Rather, the viability during addition of ?-D-glucosides in medium-free buffers is higher for strains harbouring the Bgl isoform Val204 than for those with the isoform Ala204. Conclusion Our results indicate that the studied SNP in the bgl gene is one possible cause of the difference in bacterial virulence among strains of M. mycoides subsp. mycoides SC. Bgl does not act as a direct virulence factor, but strains possessing the Bgl isoform Val204 with low hydrolysing activity are more prone to survive in environments that contain high levels of ?-D-glucosides, thus contributing in some extent to mycoplasmaemia. PMID:17439646

  11. Effect of Hominis Placenta on cutaneous wound healing in normal and diabetic mice

    PubMed Central

    Park, Ji-Yeun; Lee, Jiyoung; Jeong, Minsu; Min, Seorim; Kim, Song-Yi; Lee, Hyejung; Lim, Yunsook

    2014-01-01

    BACKGROUND/OBJECTIVES The number of diabetic patients has recently shown a rapid increase, and delayed wound healing is a major clinical complication in diabetes. In this study, the wound healing effect of Hominis placenta (HP) treatment was investigated in normal and streptozotocin-induced diabetic mice. MATERIALS/METHODS Four full thickness wounds were created using a 4 mm biopsy punch on the dorsum. HP was injected subcutaneously at the middle region of the upper and lower wounds. Wounds were digitally photographed and wound size was measured every other day until the 14th day. Wound closure rate was analyzed using CANVAS 7SE software. Wound tissues were collected on days 2, 6, and 14 after wounding for H/E, immunohistochemistry for FGF2, and Masson's trichrome staining for collagen study. RESULTS Significantly faster wound closure rates were observed in the HP treated group than in normal and diabetes control mice on days 6 and 8. Treatment with HP resulted in reduced localization of inflammatory cells in wounded skin at day 6 in normal mice and at day 14 in diabetic mice (P < 0.01). Expression of fibroblast growth factor (FGF) 2 showed a significant increase in the HP treated group on day 14 in both normal (P < 0.01) and diabetic mice (P < 0.05). In addition, HP treated groups showed a thicker collagen layer than no treatment groups, which was remarkable on the last day, day 14, in both normal and diabetic mice. CONCLUSIONS Taken together, HP treatment has a beneficial effect on acceleration of cutaneous wound healing via regulation of the entire wound healing process, including inflammation, proliferation, and remodeling. PMID:25110560

  12. 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. PMID:9577796

  13. 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. PMID:25055642

  14. Can American goldfinches function as reservoirs for Mycoplasma gallisepticum?

    PubMed

    Dhondt, André A; Dhondt, Keila V; Hochachka, Wesley M; Schat, Karel A

    2013-01-01

    We performed experiments to test if American Goldfinches (Spinus tristis) could be a competent reservoir for Mycoplasma gallisepticum and play a role in the epidemic spread of mycoplasmal conjunctivitis among House Finches (Carpodacus mexicanus) in North America. We infected one of two individuals housed together in a cage and determined if transmission occurred to the second bird. Probability of transmission between an American Goldfinch and a House Finch (in either direction) was similar to that between two House Finches. In a second experiment small groups of birds (6-8) were housed in large aviaries. Two source birds were inoculated with M. gallisepticum, and transmission to the naive birds in the aviary was recorded. Transmission occurred among House Finches, among American Goldfinches, and from House Finches to American Goldfinches. Transmission was more likely between House Finches than among American Goldfinches, and between House Finches and American Goldfinches. We conclude that American Goldfinches are a competent reservoir for Mycoplasma gallisepticum and could have played a role in the spread of the epidemic as they are more migratory than House Finches. PMID:23307371

  15. Biofunctional domains of the Mycoplasma pneumoniae P30 adhesin.

    PubMed Central

    Dallo, S F; Lazzell, A L; Chavoya, A; Reddy, S P; Baseman, J B

    1996-01-01

    The P30 adhesin genes of spontaneous, hemadsorption-negative (HA-) class II Mycoplasma pneumoniae mutants that displayed P30 adhesin-deficient protein profiles were analyzed. One subclass of P30-deficient mutants possessed the entire p3O structural gene without alterations (825 nucleotides, encoding 275 amino acids with a predicted molecular mass of 29,743 Da [S. F. Dallo, A. Chavoya, and J. B. Baseman, Infect. Immun. 58:4163-4165, 1990]). However, the second mutant subclass contained a deletion in p3O resulting in the expression of a 25-kDa peptide (681 nucleotides, encoding 227 amino acids with a calculated molecular mass of 24,823 Da). This P25-truncated peptide lacked 8 of the 13 proline-rich amino acid repeat sequences at the carboxy terminus. Whole-cell radioimmunoprecipitation of M. pneumoniae with antibodies directed against the proline-rich repeat sequences located in the carboxy terminus demonstrated their surface accessibility. In contrast, antibodies generated against N-terminal amino acid sequences upstream of the repeats did not bind to intact mycoplasmas. The amino acid sequence homologies exhibited by the P30 adhesin and eucaryotic structural proteins were corroborated by cross-reactive epitopes shared between the P30 adhesin and fibrinogen, keratin, and myosin. These data reinforce the importance of the P30 protein in cytadherence and virulence and provide a molecular basis for postinfectious autoimmunity associated with M. pneumoniae-mediated pathologies. PMID:8698484

  16. Evolutionary History of Contagious Bovine Pleuropneumonia Using Next Generation Sequencing of Mycoplasma mycoides Subsp. mycoides “Small Colony”

    PubMed Central

    Dupuy, Virginie; Manso-Silván, Lucía; Barbe, Valérie; Thebault, Patricia; Dordet-Frisoni, Emilie; Citti, Christine; Poumarat, François; Blanchard, Alain; Breton, Marc; Sirand-Pugnet, Pascal; Thiaucourt, François

    2012-01-01

    Mycoplasma mycoides subsp. mycoides “Small Colony” (MmmSC) is responsible for contagious bovine pleuropneumonia (CBPP) in bovidae, a notifiable disease to the World Organization for Animal Health (OIE). Although its origin is not documented, the disease was known in Europe in 1773. It reached nearly world-wide distribution in the 19th century through the cattle trade and was eradicated from most continents by stamping-out policies. During the 20th century it persisted in Africa, and it reappeared sporadically in Southern Europe. Yet, classical epidemiology studies failed to explain the re-occurrence of the disease in Europe in the 1990s. The objectives of this study were to obtain a precise phylogeny of this pathogen, reconstruct its evolutionary history, estimate the date of its emergence, and determine the origin of the most recent European outbreaks. A large-scale genomic approach based on next-generation sequencing technologies was applied to construct a robust phylogeny of this extremely monomorphic pathogen by using 20 representative strains of various geographical origins. Sixty two polymorphic genes of the MmmSC core genome were selected, representing 83601 bp in total and resulting in 139 SNPs within the 20 strains. A robust phylogeny was obtained that identified a lineage specific to European strains; African strains were scattered in various branches. Bayesian analysis allowed dating the most recent common ancestor for MmmSC around 1700. The strains circulating in Sub-Saharan Africa today, however, were shown to descend from a strain that existed around 1810. MmmSC emerged recently, about 300 years ago, and was most probably exported from Europe to other continents, including Africa, during the 19th century. Its diversity is now greater in Africa, where CBPP is enzootic, than in Europe, where outbreaks occurred sporadically until 1999 and where CBPP may now be considered eradicated unless MmmSC remains undetected. PMID:23071648

  17. Mutant Analysis Reveals a Specific Requirement for Protein P30 in Mycoplasma pneumoniae Gliding Motility

    PubMed Central

    Hasselbring, Benjamin M.; Jordan, Jarrat L.; Krause, Duncan C.

    2005-01-01

    The cell-wall-less prokaryote Mycoplasma pneumoniae, long considered among the smallest and simplest cells capable of self-replication, has a distinct cellular polarity characterized by the presence of a differentiated terminal organelle which functions in adherence to human respiratory epithelium, gliding motility, and cell division. Characterization of hemadsorption (HA)-negative mutants has resulted in identification of several terminal organelle proteins, including P30, the loss of which results in developmental defects and decreased adherence to host cells, but their impact on M. pneumoniae gliding has not been investigated. Here we examined the contribution of P30 to gliding motility on the basis of satellite growth and cell gliding velocity and frequency. M. pneumoniae HA mutant II-3 lacking P30 was nonmotile, but HA mutant II-7 producing a truncated P30 was motile, albeit at a velocity 50-fold less than that of the wild type. HA-positive revertant II-3R producing an altered P30 was unexpectedly not fully wild type with respect to gliding. Complementation of mutant II-3 with recombinant wild-type and mutant alleles confirmed the correlation between gliding defect and loss or alteration in P30. Surprisingly, fusion of yellow fluorescent protein to the C terminus of P30 had little impact on cell gliding velocity and significantly enhanced HA. Finally, while quantitative examination of HA revealed clear distinctions among these mutant strains, gliding defects did not correlate strictly with the HA phenotype, and all strains attached to glass at wild-type levels. Taken together, these findings suggest a role for P30 in gliding motility that is distinct from its requirement in adherence. PMID:16159760

  18. Mutant analysis reveals a specific requirement for protein P30 in Mycoplasma pneumoniae gliding motility.

    PubMed

    Hasselbring, Benjamin M; Jordan, Jarrat L; Krause, Duncan C

    2005-09-01

    The cell-wall-less prokaryote Mycoplasma pneumoniae, long considered among the smallest and simplest cells capable of self-replication, has a distinct cellular polarity characterized by the presence of a differentiated terminal organelle which functions in adherence to human respiratory epithelium, gliding motility, and cell division. Characterization of hemadsorption (HA)-negative mutants has resulted in identification of several terminal organelle proteins, including P30, the loss of which results in developmental defects and decreased adherence to host cells, but their impact on M. pneumoniae gliding has not been investigated. Here we examined the contribution of P30 to gliding motility on the basis of satellite growth and cell gliding velocity and frequency. M. pneumoniae HA mutant II-3 lacking P30 was nonmotile, but HA mutant II-7 producing a truncated P30 was motile, albeit at a velocity 50-fold less than that of the wild type. HA-positive revertant II-3R producing an altered P30 was unexpectedly not fully wild type with respect to gliding. Complementation of mutant II-3 with recombinant wild-type and mutant alleles confirmed the correlation between gliding defect and loss or alteration in P30. Surprisingly, fusion of yellow fluorescent protein to the C terminus of P30 had little impact on cell gliding velocity and significantly enhanced HA. Finally, while quantitative examination of HA revealed clear distinctions among these mutant strains, gliding defects did not correlate strictly with the HA phenotype, and all strains attached to glass at wild-type levels. Taken together, these findings suggest a role for P30 in gliding motility that is distinct from its requirement in adherence. PMID:16159760

  19. Multi-locus sequence analysis of Mycoplasma capricolum subsp. capripneumoniae for the molecular epidemiology of contagious caprine pleuropneumonia.

    PubMed

    Manso-Silván, Lucía; Dupuy, Virginie; Chu, Yuefeng; Thiaucourt, François

    2011-01-01

    Mycoplasma capricolum subsp. capripneumoniae (Mccp) is the causative agent of contagious caprine pleuropneumonia (CCPP), a devastating disease of domestic goats. The exact distribution of CCPP is not known but it is present in Africa and the Middle East and represents a significant threat to many disease-free areas including Europe. Furthermore, CCPP has been recently identified in Tajikistan and China. A typing method with an improved resolution based on Multi-Locus Sequence Analysis (MLSA) has been developed to trace new epidemics and to elucidate whether the recently identified cases in continental Asia were due to recent importation of Mccp. The H2 locus, a polymorphic region already in use as a molecular marker for Mccp evolution, was complemented with seven new loci selected according to the analysis of polymorphisms observed among the genome sequences of three Mccp strains. A total of 25 strains, including the two new strains from Asia, were analysed by MLSA resulting in the discrimination of 15 sequence types based on 53 polymorphic positions. A distance tree inferred from the concatenated sequences of the eight selected loci revealed two evolutionary lineages comprising five groups, which showed good correlation with geographic origins. The presence of a distinct Asian cluster strongly indicates that CCPP was not recently imported to continental Asia. It is more likely that the disease has been endemic in the area for a long time, as supported by historical clinical descriptions. In conclusion, this MLSA strategy constitutes a highly discriminative tool for the molecular epidemiology of CCPP. PMID:21756321

  20. Mycoplasma Contamination of Cell Cultures: Vesicular Traffic in Bacteria and Control over Infectious Agents.

    PubMed

    Chernov, V M; Chernova, O A; Sanchez-Vega, J T; Kolpakov, A I; Ilinskaya, O N

    2014-07-01

    Cell cultures are subject to contamination either with cells of other cultures or with microorganisms, including fungi, viruses, and bacteria. Mycoplasma contamination of cell cultures is of particular importance. Since cell cultures are used for the production of vaccines and physiologically active compounds, designing a system for controlling contaminants becomes topical for fundamental science and biotechnological production. The discovery of extracellular membrane vesicles in mycoplasmas makes it necessary to take into consideration the bacterial vesicular traffic in systems designed for controlling infectious agents. The extracellular vesicles of bacteria mediate the traffic of proteins and genes, participate in cell-to-cell interactions, as well as in the pathogenesis and development of resistance to antibiotics. The present review discusses the features of mycoplasmas, their extracellular vesicles, and the interaction between contaminants and eukaryotic cells. Furthermore, it provides an analysis of the problems associated with modern methods of diagnosis and eradication of mycoplasma contamination from cell cultures and prospects for their solution. PMID:25349713

  1. Opsonin-reversible resistance of Mycoplasma pneumoniae to in vitro phagocytosis by alveolar macrophages.

    PubMed Central

    Powell, D A; Clyde, W A

    1975-01-01

    Several species of mycoplasmas are responsible for respiratory disease in animals and man. As yet, little is known about the interaction of these pathogens with alveolar macrophages, one of the primary components of pulmonary resistance to infections. The present study was undertaken to develop an in vitro model to examine this organism-cell interaction, using a human pathogen, mycoplasma pneumoniae, and normal guinea pig alveolar macrophages. During a 24-h incubation of M. pneumoniae with a monolayer of macrophages, mycoplasmas were found to attach directly to the surface of the cells without inducing significant phagocytosis. Ultrastructurally, the organisms appeared bound to the cell membrane by their characteristic attachment organelles. Only after the addition of specific anti-mycoplasma serum were cells able to engulf attached and surrounding organisms. These data suggest that the interaction of M. pneumoniae and alveolar macrophages is a potentially important aspect of disease pathogenesis, and immune factors which might alter this interaction merit further examination. Images PMID:1090535

  2. Nonconserved Residues Ala287 and Ser290 of the Cryptosporidium hominis Thymidylate Synthase Domain Facilitate Its Rapid Rate of Catalysis

    SciTech Connect

    Doan,L.; Martucci, W.; Vargo, M.; Atreya, C.; Anderson, K.

    2007-01-01

    Cryptosporidium hominis TS-DHFR exhibits an unusually high rate of catalysis at the TS domain, at least 10-fold greater than those of other TS enzymes. Using site-directed mutagenesis, we have mutated residues Ala287 and Ser290 in the folate-binding helix to phenylalanine and glycine, respectively, the corresponding residues in human and most other TS enzymes. Our results show that the mutant A287F, the mutant S290G, and the double mutant all have reduced affinities for methylene tetrahydrofolate and reduced rates of reaction at the TS domain. Interestingly, the S290G mutant enzyme had the lowest TS activity, with a catalytic efficiency {approx}200-fold lower than that of the wild type (WT). The rate of conformational change of the S290G mutant is {approx}80 times slower than that of WT, resulting in a change in the rate-limiting step from hydride transfer to covalent ternary complex formation. We have determined the crystal structure of ligand-bound S290G mutant enzyme, which shows that the primary effect of the mutation is an increase in the distance between the TS ligands. The kinetic and crystal structure data presented here provide the first evidence explaining the unusually fast TS rate in C. hominis.

  3. Mycoplasmas regulate the expression of heat-shock protein genes through CIRCE–HrcA interactions

    Microsoft Academic Search

    Li-Jen Chang; Wei-Hsiuan Chen; F. Chris Minion; David Shiuan

    2008-01-01

    Mycoplasmas in general are rarely exposed to severe environmental changes except during its colonization and infection processes. Genomic analysis indicates that Mycoplasma hyopneumoniae possesses the genes of a single sigma factor and the HrcA repressor of negative regulation of the heat-shock response. A perfect inverted repeat sequence (5?-CTGGCACTT-N9-AAGTGCCAA-3?) upstream of the DnaK gene has also been identified. In the present

  4. Construction of Mini-Tn 4001tet and Its Use in Mycoplasma gallisepticum

    Microsoft Academic Search

    Ina Pour-El; Cary Adams; F. Chris Minion

    2002-01-01

    The Mollicutes are a group of cell-wall-less bacteria and are important plant and animal pathogens. Progress toward analyzing their pathogenic mechanisms has been hampered by the few available genetic tools. Of the two transposons shown to function in mycoplasmas, only Tn4001 is readily amenable to modification and development. One disadvantage of using Tn4001 in mycoplasmas has been independent insertion of

  5. Chronic Fatigue Syndrome Patients Subsequently Diagnosed with Lyme Disease Borrelia burgdorferi: Evidence for Mycoplasma Species Coinfections

    Microsoft Academic Search

    Garth L. Nicolson; Nancy L. Nicolson; Joerg Haier

    2008-01-01

    Objective: We examined the blood of 48 North American Chronic Fatigue Syndrome (CFS) patients subsequently diagnosed with Lyme Disease Borrelia burgdorferi and compared these to 50 North American CFS patients without evidence of Borrelia burgdorferi infections for presence of Mycoplasma spp. co-infections using forensic polymerase chain reaction. Results: We found that 68.75% of CFS\\/Lyme patients show evidence of mycoplasma co-infections

  6. The Effect of Multiple Evolutionary Selections on Synonymous Codon Usage of Genes in the Mycoplasma bovis Genome

    PubMed Central

    Zhou, Jian-hua; Ding, Yao-zhong; He, Ying; Chu, Yue-feng; Zhao, Ping; Ma, Li-ya; Wang, Xin-jun; Li, Xue-rui; Liu, Yong-sheng

    2014-01-01

    Mycoplasma bovis is a major pathogen causing arthritis, respiratory disease and mastitis in cattle. A better understanding of its genetic features and evolution might represent evidences of surviving host environments. In this study, multiple factors influencing synonymous codon usage patterns in M. bovis (three strains’ genomes) were analyzed. The overall nucleotide content of genes in the M. bovis genome is AT-rich. Although the G and C contents at the third codon position of genes in the leading strand differ from those in the lagging strand (p<0.05), the 59 synonymous codon usage patterns of genes in the leading strand are highly similar to those in the lagging strand. The over-represented codons and the under-represented codons were identified. A comparison of the synonymous codon usage pattern of M. bovis and cattle (susceptible host) indicated the independent formation of synonymous codon usage of M. bovis. Principal component analysis revealed that (i) strand-specific mutational bias fails to affect the synonymous codon usage pattern in the leading and lagging strands, (ii) mutation pressure from nucleotide content plays a role in shaping the overall codon usage, and (iii) the major trend of synonymous codon usage has a significant correlation with the gene expression level that is estimated by the codon adaptation index. The plot of the effective number of codons against the G+C content at the third codon position also reveals that mutation pressure undoubtedly contributes to the synonymous codon usage pattern of M. bovis. Additionally, the formation of the overall codon usage is determined by certain evolutionary selections for gene function classification (30S protein, 50S protein, transposase, membrane protein, and lipoprotein) and translation elongation region of genes in M. bovis. The information could be helpful in further investigations of evolutionary mechanisms of the Mycoplasma family and heterologous expression of its functionally important proteins. PMID:25350396

  7. Prevalence of mycoplasmas in the semen and vaginal swabs of Danish stallions and mares.

    PubMed

    Baczynska, Agata; Fedder, Jens; Schougaard, Hans; Christiansen, Gunna

    2007-03-31

    The reproduction rate of horses is one of the lowest within domestic livestock despite advances the veterinary medicine. Infertility in horses may be due mainly to the lack of suitable selection criteria in the breeding of horses. However, acquired infertility due to genital, bacterial infections may occur. Mycoplasmas have been implicated in genital disorders and infertility of many species including humans and horses. However, their role as commensals or pathogens of the genital tract of horses is still not determined. Bacteriological examinations made on the fossa glandis, urethra, penis and semen of stallions, showed the presence of different Mycoplasma species. Therefore our study aimed to find the prevalence of Mycoplasma species and a possible association with fertility problems in Danish riding horses. Eighty semen samples from stallions and 19 vaginal swab samples from mares were tested by PCR for presence of mycoplasmal DNA. The vaginal swab samples were also cultured in the Mycoplasma specific medium. None of the samples were positive for presence of genital mycoplasmas during the screen. The lack of genital mycoplasmas observed in this study may be due to a very extensive use of artificial insemination of modern sport horses. PMID:17178442

  8. Ultrafast Evolution and Loss of CRISPRs Following a Host Shift in a Novel Wildlife Pathogen, Mycoplasma gallisepticum

    PubMed Central

    Delaney, Nigel F.; Balenger, Susan; Bonneaud, Camille; Marx, Christopher J.; Hill, Geoffrey E.; Ferguson-Noel, Naola; Tsai, Peter; Rodrigo, Allen; Edwards, Scott V.

    2012-01-01

    Measureable rates of genome evolution are well documented in human pathogens but are less well understood in bacterial pathogens in the wild, particularly during and after host switches. Mycoplasma gallisepticum (MG) is a pathogenic bacterium that has evolved predominantly in poultry and recently jumped to wild house finches (Carpodacus mexicanus), a common North American songbird. For the first time we characterize the genome and measure rates of genome evolution in House Finch isolates of MG, as well as in poultry outgroups. Using whole-genome sequences of 12 House Finch isolates across a 13-year serial sample and an additional four newly sequenced poultry strains, we estimate a nucleotide diversity in House Finch isolates of only ?2% of ancestral poultry strains and a nucleotide substitution rate of 0.8?1.2×10?5 per site per year both in poultry and in House Finches, an exceptionally fast rate rivaling some of the highest estimates reported thus far for bacteria. We also found high diversity and complete turnover of CRISPR arrays in poultry MG strains prior to the switch to the House Finch host, but after the invasion of House Finches there is progressive loss of CRISPR repeat diversity, and recruitment of novel CRISPR repeats ceases. Recent (2007) House Finch MG strains retain only ?50% of the CRISPR repertoire founding (1994–95) strains and have lost the CRISPR–associated genes required for CRISPR function. Our results suggest that genome evolution in bacterial pathogens of wild birds can be extremely rapid and in this case is accompanied by apparent functional loss of CRISPRs. PMID:22346765

  9. A serine/threonine phosphatase encoded by MG_207 of Mycoplasma genitalium is critical for its virulence

    PubMed Central

    2013-01-01

    Background Bacterial signal transduction systems like two component system (TCS) and Serine/Threonine kinase (STK) and Serine/Threonine phosphatase (STP) play important roles in the virulence and pathogenesis of bacterial pathogens. Mycoplasma genitalium, a mollicute that causes the urogenital diseases urethritis and cervicitis in men and women, respectively, is a pathogen which lacks TCS but possesses STK/STP. In this study, we investigated the biochemical and virulence properties of an STP protein encoded by the gene MG_207 of this species. Results We overexpressed MG207 in Escherichia coli overexpression system as a recombinant His10MG207 protein and purified it with affinity chromatography. This recombinant protein readily hydrolyzed the substrate p-nitrophenyl phosphate (pNPP) in a dose-dependent manner. Additional studies using synthetic peptides as substrates revealed that the recombinant protein was able to hydrolyze the threonine phosphate. Further, a transposon insertion mutant strain of M. genitalium (TIM207) that lacks the protein MG207 showed differentially phosphorylated proteins when compared to the wild type G37 strain. Mass spectrometry revealed that some of the key proteins differentially phosphorylated in TIM207 strain were putative cytoskeletal protein encoded by the gene MG_328 and pyruvate dehydrogenase E1 ? chain encoded by the gene MG_274. In addition, TIM207 was noticed to be less cytotoxic to HeLa cells and this correlated with the production of less hydrogen peroxide by this strain. This strain was also less efficient in inducing the differentiation of THP-1 cell line as compared to wild type M. genitalium. Conclusions The results of the study suggest that MG207 is an important signaling protein of M. genitalium and its presence may be crucial for the virulence of this species. PMID:23432936

  10. Serological studies to determine the occurrence of Johne's disease and mycoplasma infection in the Northern-East Polish population of European bison (Bison bonasus).

    PubMed

    Krzysiak, M K; Dudek, K; Krajewska, M; Bednarek, D; Szulowski, K

    2014-01-01

    A serological study of twenty three European bison (Bison bonasus) derived from Northern-East Poland for the seroprevalence of Mycobacterium avium subsp. paratuberculosis, Mycoplasma bovis, Mycoplasma mycoides subsp. mycoides SC, Mycoplasma agalactiae and Mycoplasma capricolum subsp. capripneumoniae was conducted. Only specific antibodies to M. bovis were detected in two animals (8.7%) which were connected with the clinical signs and macroscopic anatomopathological lesions. PMID:25638988

  11. Centipede and inchworm models to explain Mycoplasma gliding.

    PubMed

    Miyata, Makoto

    2008-01-01

    The twelve Mycoplasma species known to glide on solid surfaces all lack surface flagella or pili, and no genes homologous to known motility systems have been found in the five genomes sequenced to date. Recent studies on the fastest of these species, M. mobile, examined novel proteins involved in the gliding mechanism, binding targets on the solid surfaces, energy sources and mechanical characteristics of the movements. Accordingly, I propose a working model for the gliding mechanism, called the centipede (power stroke) model, in which the 'leg' proteins repeat a cycle of binding to and release from the solid surface, using energy from ATP. Another 'inchworm model' suggested from the structural studies of a human pathogen, M. pneumoniae, is also discussed. PMID:18083032

  12. Unitary step of gliding machinery in Mycoplasma mobile

    PubMed Central

    Kinosita, Yoshiaki; Nakane, Daisuke; Sugawa, Mitsuhiro; Masaike, Tomoko; Mizutani, Kana; Miyata, Makoto; Nishizaka, Takayuki

    2014-01-01

    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 ?m?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. PMID:24912194

  13. [Neurological symptoms due to Mycoplasma pneumoniae infection in nine children].

    PubMed

    Roussel, P; Poulat, A-L; Romaszko, J-P; Labbé, A; Sarret, C

    2015-07-01

    Mycoplasma pneumoniae infection is common in children. Extrapulmonary symptoms usually reveal as neurological symptoms, mainly as encephalitis with significant morbidity and mortality. Various other neurological presentations have also been reported. We describe a cohort of nine children with neurological manifestations due to M. pneumoniae infection, including five cases of encephalitis, one of polyradiculoneuritis, one of ophthalmoplegia, one of optic neuritis, and one of myositis. Progression was variable from ad integrum recovery to severe brain damage. Diagnosis is usually confirmed by PCR and/or serological follow-up, but the latter is still insufficiently used in practice to systematically affirm the diagnosis. Therapeutic management is not clearly defined and long-term progression can be uncertain despite early antibiotic and/or anti-inflammatory treatments. PMID:26047743

  14. Wildlife surveillance during a Mycoplasma gallisepticum epornitic in domestic turkeys.

    PubMed

    Stallknecht, D E; Johnson, D C; Emory, W H; Kleven, S H

    1982-01-01

    During a major Mycoplasma gallisepticum (MG) epornitic in domestic turkeys, tracheal swabs were collected and cultured from 477 and 770 potentially exposed wild mammals and birds, respectively. All culture attempts were negative. Serum-plate (SP) and hemagglutination-inhibition (HI) tests on 770 bird sera revealed low titers (less than or equal to 1:40) in 0.9% of tested house sparrows, 1.1% of brown-headed cowbirds, 35.7% of common grackles, 1.0% of starlings, and 16.6% of eastern meadowlarks. Low titers are believed to have resulted from birds feeding on contaminated litter and becoming sensitized. Wildlife species did not appear to be involved in transmission or maintenance of MG but may have been mechanical carriers of this pathogen. PMID:7159324

  15. Detection of Mycoplasma agassizii in the Texas Tortoise (Gopherus berlandieri)

    USGS Publications Warehouse

    Guthrie, Amanda L.; White, C. LeAnn; Brown, Mary B.; deMaar, Thomas W.

    2013-01-01

    Mycoplasma agassizii causes upper respiratory tract disease (URTD) in Texas tortoises (Gopherus berlandieri). To determine exposure to and shedding of M. agassizii, we collected blood samples and nasal swabs from 40 free-ranging Texas tortoises on public and private lands in Texas, USA, from May to October 2009. We used an enzyme-linked immunosorbent assay (ELISA) to detect M. agassizii–specific antibodies. Eleven (28%) tortoises were antibody positive, three (8%) were suspect, and the remaining 26 (65%) were negative. Nasal lavage samples were collected from 35 of the 40 tortoises for M. agassizii culture and PCR to detect shedding of M. agassizii. Current infection with M. agassizii was confirmed in one tortoise that had mild clinical signs of URTD and was positive by ELISA (antibody titer >512), PCR, and culture. The clinical isolate was confirmed as M. agassizii by restriction fragment length polymorphism and immunobinding.

  16. Mycoplasma corogypsi associated polyarthritis and tenosynovitis in black vultures (Coragyps atratus)

    PubMed Central

    Van Wettere, A. J.; Ley, D. H.; Scott, D. E.; Buckanoff, H. D.; Degernes, L. A.

    2013-01-01

    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. PMID:22903399

  17. Macrolide-resistant Mycoplasma pneumoniae in adolescents with community-acquired pneumonia

    PubMed Central

    2012-01-01

    Background Although the prevalence of macrolide-resistant Mycoplasma pneumoniae isolates in Japanese pediatric patients has increased rapidly, there have been no reports concerning macrolide-resistant M. pneumoniae infection in adolescents aged 16 to 19 years old. The purpose of this study was to clarify the prevalence and clinical characteristics of macrolide-resistant M. pneumoniae in adolescent patients with community-acquired pneumonia. Methods A total of 99 cases with M. pneumoniae pneumonia confirmed by polymerase chain reaction (PCR) and culture were analyzed. Forty-five cases were pediatric patients less than 16 years old, 26 cases were 16 to 19-year-old adolescent patients and 28 cases were adult patients. Primers for domain V of 23S rRNA were used and DNA sequences of the PCR products were compared with the sequence of an M. pneumoniae reference strain. Results Thirty of 45 pediatric patients (66%), 12 of 26 adolescent patients (46%) and seven of 28 adult patients (25%) with M. pneumoniae pneumonia were found to be infected with macrolide-resistant M. pneumoniae (MR patients). Although the prevalence of resistant strains was similar in pediatric patients between 2008 and 2011, an increase in the prevalence of resistant strains was observed in adolescent patients. Among 30 pediatric MR patients, 26 had an A-to-G transition at position 2063 (A2063G) and four had an A-to-G transition at position 2064 (A2064G). In 12 adolescent MR patients, 10 showed an A2063G transition and two showed an A2064G transition, and in seven adult MR patients, six showed an A2063G transition and one showed an A2064G transition. Conclusions The prevalence of macrolide-resistant M. pneumoniae is high among adolescent patients as well as pediatric patients less than 16-years old. To prevent outbreaks of M. pneumoniae infection, especially macrolide-resistant M. pneumoniae, in closed populations including among families, in schools and in university students, physicians should pay close attention to macrolide-resistant M. pneumoniae. PMID:22650321

  18. Intestinal protozoa and helminths among Terena Indians in the State of Mato Grosso do Sul: high prevalence of Blastocystis hominis.

    PubMed

    Aguiar, José Ivan Albuquerque; Gonçalves, Alessandra Queiroga; Sodré, Fernando Campos; Pereira, Severino Dos Ramos; Bóia, Márcio Neves; de Lemos, Elba Regina Sampaio; Daher, Roberto Ruhman

    2007-01-01

    A parasitological survey was carried out among Terena Indians living in the Tereré settlement in the municipality of Sidrolândia, State of Mato Grosso do Sul, Brazil. Single samples of feces from 313 Indians were processed by means of the spontaneous sedimentation method. In the population studied, 73.5% were infected with at least one intestinal parasite or commensal. Protozoa predominated. Blastocystis hominis (40.9%), Entamoeba coli (33.2%) and Entamoeba histolytica/Entamoeba dispar (31.6%) were the most common. Bivariate analysis showed that females were generally more infected and presented higher rates of infection by Entamoeba histolytica/Entamoeba dispar and Entamoeba coli. Males were more infected by hookworms and Strongyloides stercoralis than females. The precarious sanitary conditions of the Tereré settlement are probably a contributory factor towards the high prevalence of intestinal protozoa. PMID:18200414

  19. Mycoplasma genitalium Infection Is Associated with Microscopic Signs of Cervical Inflammation in Liquid Cytology Specimens

    PubMed Central

    Dehon, Patricia M.

    2014-01-01

    Cervicitis is a common clinical finding often attributed to sexually transmitted infections (STIs), but no etiologic agent is identified in the majority of cases. In this study, we comparatively assessed inflammation among the common infectious etiologies of cervicitis and assessed the potential value of liquid cytology specimens for predicting STIs. Among 473 Louisiana women at low risk for acquiring STIs, the prevalences of Mycoplasma genitalium, Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis in liquid-based cytology specimens were 1.5, 2.1, 0.6, and 4.4%, respectively. N. gonorrhoeae and human papillomavirus 18 (HPV18) infections were significantly more common among subjects infected with M. genitalium. Using direct microscopy, we observed significant increases in leukocyte infiltrates among subjects with monoinfections with M. genitalium or C. trachomatis compared to women with no detectable STIs. Inflammation was highest among subjects with M. genitalium. Using a threshold of ?2 leukocytes per epithelial cell per high-powered field, the positive predictive values for M. genitalium, C. trachomatis, N. gonorrhoeae, and T. vaginalis were 100, 70, 67, and 20%, respectively. Several novel M. genitalium genotypes were identified, all of which were predicted to be susceptible to macrolide antibiotics, suggesting that different strains may circulate among low-risk women and that macrolide resistance is substantially lower than in high-risk populations. This study highlights the capacity of M. genitalium to elicit cervical inflammation and, considering the strong epidemiologic associations between M. genitalium and human immunodeficiency virus (HIV), provides a potential mechanism for acquisition and shedding of HIV via chronic leukocyte recruitment to the cervical mucosa. PMID:24759719

  20. Molecular evidence for hemotropic Mycoplasma infection in a Japanese badger (Meles meles anakuma) and a raccoon dog (Nyctereutes procyonoides viverrinus).

    PubMed

    Harasawa, Ryô; Orusa, Riccardo; Giangaspero, Massimo

    2014-04-01

    We report detection of hemoplasma in wild Japanese badgers (Meles meles anakuma) and raccoon dogs (Nyctereutes procyonoides viverrinus). Sequence analysis of the entire 16S rRNA genes identified Mycoplasma haemocanis in the raccoon dog sample, and a potential novel Mycoplasma species in the Japanese badger. PMID:24484489

  1. Mycoplasma hyopneumoniae mhp379 Is a Ca2+Dependent, Sugar-Nonspecific Exonuclease Exposed on the Cell Surface

    Microsoft Academic Search

    Jonathan A. Schmidt; Glenn F. Browning; Philip F. Markham

    2007-01-01

    Mycoplasma hyopneumoniae mhp379 is a putative lipoprotein that shares significant amino acid sequence similarity with a family of bacterial thermostable nucleases. To examine the nuclease activity of mhp379, the gene was cloned and expressed in Escherichia coli following the deletion of the amino-terminal signal sequence and prokaryotic lipoprotein cleavage site and mutagenesis of the mycoplasma TGA tryptophan codons to TGG.

  2. Role of Mycoplasma genitalium MG218 and MG317 cytoskeletal proteins in terminal organelle organization, gliding motility and cytadherence

    Microsoft Academic Search

    Oscar Q. Pich; Raul Burgos; Mario Ferrer-Navarro; Enrique Querol; Jaume Pinol

    2008-01-01

    The terminal organelle is a differentiated structure that plays a key role in mycoplasma cytadherence and locomotion. For this reason, the analysis of Mycoplasma genitalium mutants displaying anomalous terminal organelles could improve our knowledge regarding the structural elements required for proper locomotion. In this study, we isolated several M. genitalium mutants having transposon insertions within the mg218 or mg317 genes,

  3. Prevalence of human papillomavirus infection in the oropharynx and urine among sexually active men: a comparative study of infection by papillomavirus and other organisms, including Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma spp., and Ureaplasma spp

    PubMed Central

    2014-01-01

    Background Oropharyngeal squamous cell carcinoma (OSCC) has shown a gradual increase in male predominance due to the increasing incidence of human papillomavirus (HPV)-associated OSCC. However, the mode of HPV transmission to the oral cavity is poorly understood, and little is known about the epidemiology of oral HPV infection in men. The prevalence rates of HPV, Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma spp., and Ureaplasma spp. were compared in the oropharynx (oral cavity) and urine of male Japanese patients attending a sexually transmitted disease clinic. Methods The study population consisted of 213 men aged 16?–?70 years old (mean: 34.4 years old). Oropharyngeal gargles and urine were collected, and sedimented cells were preserved in liquid-based cytology solution. After DNA extraction, ?-globin and infectious organisms were analyzed by a PCR-based method. The HPV genotype was determined by HPV GenoArray test. Results ?-Globin was positive in 100% and 97.7% of oral and urine samples, respectively. HPV detection rates were 18.8% and 22.1% in oral and urine samples, respectively, suggesting that the prevalence of HPV infection in the oral cavity was similar to that in the urinary tract. N. gonorrhoeae was more prevalent in oral (15.6%) than urine samples (9.1%), whereas C. trachomatis was detected more frequently in urine (15.9%) than oral samples (4.2%). The detection rates of M. genitalium, M. hominis, and Ureaplasma spp. were 5.2%, 10.3%, and 16.0% in oral samples, and 7.7%, 6.3%, and 19.2% in urine, respectively. There were no significant differences in the detection rates of Mycoplasma spp. and Ureaplasma spp. between anatomical locations. The distribution of HPV types were similar in oral and urine samples, and HPV16 was the most common type. The majority of men with HPV infection in both the oral cavity and urine had concordant oral and urinary HPV infection. The presence of urinary HPV infection was an independent risk factor of oral HPV infection, with an odds ratio of 3.39 (95% CI: 1.49?–?7.71), whereas oral gonococcal infection was inversely correlated with oral HPV infection (odds ratio: 0.096; 95% CI: 0.01?–?0.77). Conclusions Oral HPV infection commonly occurs in sexually active men, and is significantly correlated with urinary HPV infection. PMID:24468054

  4. Effect of Mycoplasma synoviae and lentogenic Newcastle disease virus coinfection on cytokine and chemokine gene expression in chicken embryos.

    PubMed

    Bolha, Luka; Bencina, Dusan; Cizelj, Ivanka; Oven, Irena; Slavec, Brigita; Rojs, Olga Zorman; Narat, Mojca

    2013-12-01

    Mycoplasma synoviae and Newcastle disease virus (NDV) are 2 avian pathogens that cause modulation in expression of a variety of cytokine and chemokine genes in chickens. However, there is limited data about gene modulation after coinfection with these 2 pathogens and even less data about gene modulation after infection of chicken embryos. In this study, the effect of M. synoviae type strain WVU 1853 and lentogenic LaSota vaccine strain of NDV infection on cytokine and chemokine gene expression in chicken embryos was analyzed in the liver, spleen, bursa of Fabricius, and thymus by using quantitative real-time PCR. Three types of infection were performed; infection with M. synoviae on d 10, infection with NDV on d 17; and consecutive infection with both pathogens, where M. synoviae was inoculated on d 10 and NDV on d 17. Thus, simulation of consecutive infection that may occur after NDV infection of the M. synoviae-infected host was performed. Mycoplasma synoviae infection of embryos resulted in intensive upregulation of cytokine and chemokine genes, including interferon (IFN)-?, IL-1?, IL-6, IL-12p40, IL-16, IL-18, MIP-1? (CCL4), inducible nitric oxide synthase (iNOS), XCL1, and lipopolysaccharide-induced tumor necrosis factor-? factor (LITAF), with different expression profiles in the 4 organs. Inoculation of lentogenic NDV significantly upregulated IFN-?, IL-6, and IL-16 genes in spleen and IFN-?, IL-1?, IL-2, IL-16, IL-21, XCL1, and MIP-1? (CCL4) genes in the thymus, but to a lesser extent than M. synoviae. However, no genes were upregulated by NDV in the liver and bursa of Fabricius. Overall effect of NDV inoculation, regarding the number of modulated cytokine and chemokine genes and the extent of expression, was lower than M. synoviae. When NDV was introduced after on-going M. synoviae infection, most M. synoviae-induced cytokine and chemokine genes were significantly downregulated. This study provides the first evidence in chicken embryos that consecutive infection with NDV could suppress expression of cytokine and chemokine genes being significantly upregulated by the previous M. synoviae infection. PMID:24235222

  5. Mycoplasma infection suppresses p53, activates NF-kappaB and cooperates with oncogenic Ras in rodent fibroblast transformation.

    PubMed

    Logunov, D Y; Scheblyakov, D V; Zubkova, O V; Shmarov, M M; Rakovskaya, I V; Gurova, K V; Tararova, N D; Burdelya, L G; Naroditsky, B S; Ginzburg, A L; Gudkov, A V

    2008-07-31

    Prokaryotes of the genus Mycoplasma are the smallest cellular organisms that persist as obligate extracellular parasites. Although mycoplasma infection is known to be associated with chromosomal instability and can promote malignant transformation, the mechanisms underlying these phenomena remain unknown. Since persistence of many cellular parasites requires suppression of apoptosis in host cells, we tested the effect of mycoplasma infection on the activity of the p53 and nuclear factor (NF)-kappaB pathways, major mechanisms controlling programmed cell death. To monitor the activity of p53 and NF-kappaB in mycoplasma-infected cells, we used a panel of reporter cell lines expressing the bacterial beta-galactosidase gene under the control of p53- or NF-kappaB-responsive promoters. Cells incubated with media conditioned with different species of mycoplasma showed constitutive activation of NF-kappaB and reduced activation of p53, common characteristics of the majority of human tumor cells, with M. arginini having the strongest effect among the species tested. Moreover, mycoplasma infection reduced the expression level and inducibility of an endogenous p53-responsive gene, p21(waf1), and inhibited apoptosis induced by genotoxic stress. Infection with M. arginini made rat and mouse embryo fibroblasts susceptible to transformation with oncogenic H-Ras, whereas mycoplasma-free cells underwent irreversible p53-dependent growth arrest. Mycoplasma infection was as effective as shRNA-mediated knockdown of p53 expression in making rodent fibroblasts permissive to Ras-induced transformation. These observations indicate that mycoplasma infection plays the role of a p53-suppressing oncogene that cooperates with Ras in cell transformation and suggest that the carcinogenic and mutagenic effects of mycoplasma might be due to inhibition of p53 tumor suppressor function by this common human parasite. PMID:18408766

  6. Effects of Mycoplasma gallisepticum vaccination on serum ?1-acid glycoprotein concentrations in commercial layer chickens.

    PubMed

    Peebles, E D; Jacob, R; Branton, S L; Gerard, P D

    2014-06-01

    Increases in circulating acute phase protein (APP) levels occur in reaction to systemic infections in animals. However, no previous research has been conducted to monitor possible changes in APP levels of birds in response to prelay vaccinations of various live attenuated Mycoplasma gallisepticum vaccines in conjunction with their subsequent use as an overlay vaccine during the production period. Serum concentrations of the APP, ?1-acid glycoprotein (AGP), were determined on d 0, 1, 3, 7, 14, and 28 after subjecting commercial laying hens to one of the following treatments at 10 wk of age (woa): 1) control (no vaccination); 2) ts-11 strain M. gallisepticum (ts11MG) vaccination; 3) M. gallisepticum-bacterin (MGBac) vaccination; and 4) ts11MG and MGBac combination (ts11MG & MGBac) vaccination. Furthermore, at 45 woa, the birds in half of the units assigned to each treatment group were vaccinated with high-passage F-strain M. gallisepticum (HpFMG). Birds in treatment 1 that were (single control) and were not (double control) vaccinated with HpFMG, and birds in treatments 2, 3, and 4 that were vaccinated with HpFMG were further tested during lay on d 0, 1, 3, 7, 14, and 28 after vaccination. On d 7, 14, and 28 postvaccination at 10 woa, the ts11MG & MGBac, ts11MG, and MGBac group AGP concentrations were not different from one another, but all were higher than those in the control group. Similarly, on d 3, 7, and 14 postvaccination, the single control, and the MGBac ts11MG, and ts11MG & MGBac treatment groups that were later vaccinated with HpFMG at 45 woa, were not different, but all were higher than that in the double control group. In conclusion, elevated circulation AGP concentrations may be used to detect and confirm subclinical infections in pullets up to 28 d after having been vaccinated with ts11MG, MGBac, or their combination. Furthermore, in association with depressed performance, elevated serum AGP concentrations in layers may be used to confirm HpFMG infections up to 28 d after its use as a vaccine. PMID:24879689

  7. A Sialoreceptor Binding Motif in the Mycoplasma synoviae Adhesin VlhA

    PubMed Central

    May, Meghan; Dunne, Dylan W.; Brown, Daniel R.

    2014-01-01

    Mycoplasma synoviae depends on its adhesin VlhA to mediate cytadherence to sialylated host cell receptors. Allelic variants of VlhA arise through recombination between an assemblage of promoterless vlhA pseudogenes and a single transcription promoter site, creating lineages of M. synoviae that each express a different vlhA allele. The predicted full-length VlhA sequences adjacent to the promoter of nine lineages of M. synoviae varying in avidity of cytadherence were aligned with that of the reference strain MS53 and with a 60-a.a. hemagglutinating VlhA C-terminal fragment from a Tunisian lineage of strain WVU1853T. Seven different sequence variants of an imperfectly conserved, single-copy, 12-a.a. candidate cytadherence motif were evident amid the flanking variable residues of the 11 total sequences examined. The motif was predicted to adopt a short hairpin structure in a low-complexity region near the C-terminus of VlhA. Biotinylated synthetic oligopeptides representing four selected variants of the 12-a.a. motif, with the whole synthesized 60-a.a. fragment as a positive control, differed (P<0.01) in the extent they bound to chicken erythrocyte membranes. All bound to a greater extent (P<0.01) than scrambled or irrelevant VlhA domain negative control peptides did. Experimentally introduced branched-chain amino acid (BCAA) substitutions Val3Ile and Leu7Ile did not significantly alter binding, whereas fold-destabilizing substitutions Thr4Gly and Ala9Gly tended to reduce it (P<0.05). Binding was also reduced to background levels (P<0.01) when the peptides were exposed to desialylated membranes, or were pre-saturated with free sialic acid before exposure to untreated membranes. From this evidence we conclude that the motif P-X-(BCAA)-X-F-X-(BCAA)-X-A-K-X-G binds sialic acid and likely mediates VlhA-dependent M. synoviae attachment to host cells. This conserved mechanism retains the potential for fine-scale rheostasis in binding avidity, which could be a general characteristic of pathogens that depend on analogous systems of antigenically variable adhesins. The motif may be useful to identify previously unrecognized adhesins. PMID:25338071

  8. Update on Pneumocystis carinii f. sp. hominis Typing Based on Nucleotide Sequence Variations in Internal Transcribed Spacer Regions of rRNA Genes

    Microsoft Academic Search

    CHAO-HUNG LEE; JANNIK HELWEG-LARSEN; XING TANG; SHAOLING JIN; BAOZHENG LI; MARILYN S. BARTLETT; JANG-JIH LU; BETTINA LUNDGREN; JENS D. LUNDGREN; MATS OLSSON; SEBASTIAN B. LUCAS; PATRICIA ROUX; ANTONIETTA CARGNEL; CHIARA ATZORI; OLGA MATOS; JAMES W. SMITH; Ospedale L. Sacco

    1998-01-01

    Pneumocystis carinii f. sp. hominis isolates from 207 clinical specimens from nine countries were typed based on nucleotide sequence variations in the internal transcribed spacer regions I and II (ITS1 and ITS2, respectively) of rRNA genes. The number of ITS1 nucleotides has been revised from the previously reported 157 bp to 161 bp. Likewise, the number of ITS2 nucleotides has

  9. Lesson of the month 1: Pericardial mass and cardiac tamponade associated with Mycoplasma pneumoniae.

    PubMed

    Sawhney, Vinit; Maksunova, Oksana; Ahsan, Syed; Ozkor, Muhiddin; Westwood, Mark

    2014-10-01

    Mycoplasma pneumoniae primarily causes respiratory tract infections. Extrapulmonary manifestations are seen in 20-25% of cases. Cardiac involvement is rarely reported. We present a unique case of a pericardial mass and cardiac tamponade associated with a Mycoplasma pneumoniae pneumonia. This required emergency pericardial fenestration. The patient improved dramatically postoperatively on antibiotics and there was no recurrence of the pericardial effusion on follow up. This case highlights the often forgotten invasive properties of a common respiratory tract pathogen and emphasises the need to consider this easily treatable entity in the differential diagnosis of idiopathic pericardial effusions. PMID:25301923

  10. 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.

  11. Intracellular structures of Mycoplasma pneumoniae revealed after membrane removal.

    PubMed

    Meng, K E; Pfister, R M

    1980-10-01

    Mycoplasma pneumoniae was grown on Formvar- and carbon-coated electron microscope grids and treated with the nonionic detergent Triton X-100 to gently remove the membrane and cytoplasm. The detergent mixture was composed of 0.5% Triton X-100 in SSR-2 broth base. After this treatment, the grids were rinsed in a mixture of 0.1 M KCl, 5 mM MgCl2, and 6 mM potassium phosphate buffer (pH 7.05) and negatively stained with uranyl acetate. The Triton X-100-resistant remains of M. pneumoniae after gentle removal of the membrane and cytoplasm consisted of fibrous structures oriented similarly to the undisrupted cells. The thin fibers displayed a negative staining quality and diameter analogous to that of rabbit muscle F-actin. The fibrous moieties ended in rodlike condensations which appeared striated in negatively stained and shadowed preparations. These striations were regular, and the majority of rod structures had lengths of 220 to 300 nm and widths of 50 to 80 nm. Specific antibody to rabbit muscle actin, produced in guinea pigs, was used in indirect immunofluorescence of the M. pneumoniae colonies. Fluorescence was detected, with concentrations at the colony center and at the tips of filamentous cells. PMID:6774963

  12. Mycoplasma pneumoniae and Its Role as a Human Pathogen

    PubMed Central

    Waites, Ken B.; Talkington, Deborah F.

    2004-01-01

    Mycoplasma pneumoniae is a unique bacterium that does not always receive the attention it merits considering the number of illnesses it causes and the degree of morbidity associated with it in both children and adults. Serious infections requiring hospitalization, while rare, occur in both adults and children and may involve multiple organ systems. The severity of disease appears to be related to the degree to which the host immune response reacts to the infection. Extrapulmonary complications involving all of the major organ systems can occur in association with M. pneumoniae infection as a result of direct invasion and/or autoimmune response. The extrapulmonary manifestations are sometimes of greater severity and clinical importance than the primary respiratory infection. Evidence for this organism's contributory role in chronic lung conditions such as asthma is accumulating. Effective management of M. pneumoniae infections can usually be achieved with macrolides, tetracyclines, or fluoroquinolones. As more is learned about the pathogenesis and immune response elicited by M. pneumoniae, improvement in methods for diagnosis and prevention of disease due to this organism may occur. PMID:15489344

  13. THE MULTIFACETED ROLE OF T CELL-MEDIATED IMMUNITY IN PATHOGENESIS AND RESISTANCE TO MYCOPLASMA RESPIRATORY DISEASE

    PubMed Central

    Dobbs, Nicole A.; Odeh, Adam N.; Sun, Xiangle; Simecka, Jerry W.

    2010-01-01

    Mycoplasma respiratory diseases have a significant impact on the economy, health and wildlife. The hallmark of these diseases is the persistence of the mycoplasma infections and chronic inflammatory responses associated with the airways. There is still much that needs to be understood about the immune mechanisms involved in mycoplasma disease and resistance from infection. It is clear that immune responses can contribute to the generation of inflammatory lesions in mycoplasma respiratory disease, as well as provide protection from infection and extrapulmonary dissemination of the organisms. The evolution of this lung disease is under the control innate immune mechanisms and the contrasting effects of different T cell populations. The mechanisms of immunity involved in mycoplasma diseases are multifaceted, and a fascinating story of its complexity is being uncovered. Research in mycoplasma respiratory diseases have underscored the idea that immunity along the respiratory tract against infectious agents is a dynamic process and involves a network of cellular and cytokine signals that determine the type of responses generated, and ultimately, the outcome of infection. The aim of this article is to present on overview of our work on mycoplasma disease and immunity, focusing on the interactions and regulation of T cell responses that influence disease pathogenesis. We will first provide an overview of immune mechanisms involved in controlling infection and participate in the generation of T cell responses, and the role of T cell populations in generating protection and contributing to lesion development will be discussed. PMID:21743780

  14. Management practices associated with the bulk tank milk prevalence of Mycoplasma spp. in dairy herds in Northwestern Portugal.

    PubMed

    Pinho, L; Thompson, G; Machado, M; Carvalheira, J

    2013-01-01

    The objective of this study was to evaluate the effect of some management practices on the prevalence of Mycoplasma spp. in Northwestern Portuguese dairy farms from bulk tank milk (BTM) samples. Additionally, the within-herd prevalence of Mycoplasma spp. was also determined, but only in BTM positive herds. From May 2007 to November 2008, 492 BTM samples from 164 dairies randomly chosen in a population of 1234 dairy farms were analyzed. Five herds (3.0%) had positive mycoplasmal culture results, from which 4 out of 164 (2.4%) were Mycoplasma bovis, with simultaneous presence of Mycoplasma bovigenitalium or Mycoplasma canadense in two of those samples. In one out of 164 (0.6%) herds Mycoplasma capricolum subsp. capricolum was also found. In BTM positive Mycoplasma spp. herds, the apparent intra-herd prevalence was low and varied between 2.5% and 4.5%. Multiple locus variable-number of tandem-repeat analysis was conducted in order to compare the genetic relationship between the isolates. Mycoplasma spp. was found to be present in cows with subclinical mastitis with or without California Mastitis Test positive results, hence all cows should be tested when the agent is isolated from bulk tank rather than selecting suspected cows. A multivariable logistic regression using the Firth's penalized maximum likelihood estimation was performed showing that increasing number of lactating cows (OR=1.05; P<0.01) was associated with a higher probability of isolating Mycoplasma spp. On the other hand, identifying problem cows was associated with a lower probability (OR=0.06; P<0.05). Particular importance was given to the prevalence of M. bovis, and the results obtained highlight the need to include this agent in mastitis control protocols in national dairies and in sanitary controls of transitioned animals between European countries. PMID:22836035

  15. R1 Region of P97 Mediates Adherence of Mycoplasma hyopneumoniae to Swine Cilia

    Microsoft Academic Search

    F. CHRIS MINION; CARY ADAMS; TSUNGDA HSU

    2000-01-01

    Adherence of Mycoplasma hyopneumoniae to the swine respiratory tract is mediated by the membrane protein P97. This protein is located on the outer membrane surface, and its role in adherence has been firmly established. The general region of P97 that mediates adherence to swine cilia is thought to be the R1 region near the carboxy terminus of the protein, but

  16. In Vivo Expression Analysis of the P97 and P102 Paralog Families of Mycoplasma hyopneumoniae

    Microsoft Academic Search

    Cary Adams; Joshua Pitzer; F. C. Minion

    2005-01-01

    Mycoplasma hyopneumoniae, the agent of enzootic pneumo- nia of pigs, is found throughout the world, including both third world and industrialized countries (8). In addition to the chronic disease it causes, recent evidence also suggests that M. hyopneumoniae contributes significantly to the pathogenesis of other infectious agents (10). The mechanism(s) it employs to cause disease is poorly understood, but it

  17. Global transcriptional analysis of Mycoplasma hyopneumoniae following exposure to hydrogen peroxide

    Microsoft Academic Search

    Erin R. Schafer; Michael J. Oneal; Melissa L. Madsen; F. Chris Minion

    2007-01-01

    Mycoplasma hyopneumoniae, the causative agent of swine enzootic pneumonia, colonizes the cilia of swine lungs, causing ciliostasis and cell death. M. hyopneumoniae is a component of the porcine respiratory disease complex (PRDC) and is especially problematic for the finishing swine industry, causing the loss of hundreds of millions of dollars in farm revenues worldwide. For successful infection, M. hyopneumoniae must

  18. A multilocus sequence typing method and curated database for Mycoplasma bovis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis is a primary agent of mastitis, pneumonia and arthritis in cattle and is the bacterium isolated most frequently from the polymicrobial syndrome known as bovine respiratory disease complex (BRDC). Recently, M. bovis has emerged as a significant problem in bison, causing necrotic pha...

  19. Comparison of Mycoplasma agalactiae isolates by pulsed field gel electrophoresis, SDS-PAGE and immunoblotting

    Microsoft Academic Search

    Sebastiana Tola; Graziano Idini; Daniela Manunta; Ida Casciano; angela M. Rocchigiani; Antonio Angioi; Guido Leori

    1996-01-01

    We have analyzed 81 isolates of Mycoplasma agalactiae from four different regions of Italy between 1990 and 1995 in order to identify antigenic differences through SDS-PAGE and Western blotting and chromosomal DNA restriction endonuclease cleavage pattern differences. Antigenic variability in M. agalactiae isolates was investigated analyzing hydrophobic membrane protein fractions by immunoblotting using pooled sheep antiserum from naturally infected sheep.

  20. Necrotic pharyngitis associated with Mycoplasma bovis infections in American bison (Bison bison)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis (M. bovis) has recently emerged as a significant and costly infectious disease problem in bison, generally presenting as severe, caseonecrotic pneumonia. Here we describe three diagnostic cases in which M. bovis is strongly implicated as a causative agent of necrotic pharyngitis. ...

  1. Mycoplasma pneumoniae pneumonia revisited within the German Competence Network for Community-acquired pneumonia (CAPNETZ)

    Microsoft Academic Search

    Heike von Baum; Tobias Welte; Reinhard Marre; Norbert Suttorp; Christian Lück; Santiago Ewig

    2009-01-01

    BACKGROUND: Currently, broad empiric antimicrobial treatment including atypical coverage is recommended for patients with mild to moderate community-acquired pneumonia (CAP). Therefore, the relative impact of each atypical pathogen, particularly Mycoplasma pneumoniae deserves renewed attention. METHODS: Based on prospective data from 4532 patients with CAP included in the German CAP-Competence Network (CAPNETZ), we studied the incidence, clinical characteristics, and outcome of

  2. Induction of eggshell apex abnormalities by Mycoplasma synoviae: field and experimental studies

    Microsoft Academic Search

    A. Feberwee; J. J. de Wit; W. J. M. Landman

    2009-01-01

    A novel eggshell pathology, characterized by an altered shell surface, thinning, increased translucency, and cracks and breaks in the eggshell apex, has become increasingly common in layer flocks of various breeds in The Netherlands. Two field studies found an association between the eggshell apex abnormalities (EAA) and infection with Mycoplasma synoviae. M. synoviae was isolated from the oviduct of birds

  3. Cold Agglutinins in HIV-Seropositive Participants and Diagnosis of Respiratory Disease Due to Mycoplasma pneumoniae

    Microsoft Academic Search

    Esaki Muthu Shankar; Ramachandran Vignesh; Pachamuthu Balakrishnan; Vijayakumar Velu; Esakimuthu Ponmalar; Kailapuri G. Murugavel; Shanmugam Saravanan; Panneerselvam Nandagopal; Kownhar Hayath; Suniti Solomon; Appasamy Vengatesan; Usha Anand Rao

    2009-01-01

    Objectives. Cold agglutinin (CA) titers are one among the first pathological indicators for diagnosing Mycoplasma pneumoniae disease. We prospectively studied the prevalence of CAs in 300 HIV-positive and 75 HIV-negative individuals with respiratory disease in Chennai, India. Methods. The cold agglutination test was used and retrospectively compared with the results of a particle agglutination test. Results. While CAs were positive

  4. Morphology, morphometry and electron microscopy of HeLa cells infected with bovine Mycoplasma

    Microsoft Academic Search

    Edwin Boatman; Frank Cartwright; George Kenny

    1976-01-01

    The host-parasite relationship of HeLa M cells artificially infected with a bovine species of Mycoplasma was studied by light microscopy, transmission electron microscopy and scanning electron microscopy. The use of morphometry to quantitate some of the findings was explored. The parasites were seen in locations extracellular to the cell surface. The detection of small numbers of organisms by light microscopy

  5. Mycoplasma ovipneumoniae can predispose bighorn sheep to fatal Mannheimia haemolytica pneumonia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma ovipneumoniae has been isolated from the lungs of pneumonic bighorn sheep (BHS). However experimental reproduction of fatal pneumonia in BHS with M. ovipneumoniae was not successful. Therefore the specific role, if any, of M. ovipneumoniae in BHS pneumonia is unclear. The objective of th...

  6. Abortion associated with Mycoplasma bovis (M. bovis) in a bison (Bison bison) herd

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis (M. bovis) has recently emerged as a significant health threat in bison and is an increasing concern and source of economic loss for producers. Clinical manifestations of infection documented in bison include pneumonia, respiratory distress and polyarthritis. The current study des...

  7. Preserved goat milk as a valid sample for the PCR detection of Mycoplasma agalactiae

    Microsoft Academic Search

    Joaquín Amores; Christian de la Fe; Ángel Gómez-Martín; Juan C. Corrales; Antonio Contreras; Antonio Sánchez

    2011-01-01

    Goat milk samples routinely obtained by milk distributing companies and quality control laboratories are preserved using azidiol (AZ) or bronopol (BR). This study was designed to determine if these treated goat milk samples are suitable for Mycoplasma (M.) agalactiae detection by PCR. The effects of these preservatives on the limits of M. agalactiae direct PCR detection were established using samples

  8. A Genome-Scale Metabolic Reconstruction of Mycoplasma genitalium, iPS189

    E-print Network

    Maranas, Costas

    for the construction and curation of a genome-scale in silico metabolic model for M. genitalium. Key challenges sequenced [13] whereas only about 20 organism-specific genome- scale metabolic models have been constructedA Genome-Scale Metabolic Reconstruction of Mycoplasma genitalium, iPS189 Patrick F. Suthers1

  9. The response of chickens to experimental infection ‘in ovo’ with Mycoplasma synoviae

    Microsoft Academic Search

    Janet M. Bradbury; L. J. Howell

    1975-01-01

    Embryonated chicken eggs were inoculated at 18 days of incubation with a broth culture of Mycophsma synoviae and the infected chicks were hatched and reared in isolation. Control chicks were hatched following inoculation of eggs with sterile mycoplasma broth.Both groups were closely observed for clinical signs and, at intervals up to 6 weeks infected and control chicks were killed and

  10. Multilocus sequence typing of Mycoplasma bovis reveals host-specific genotypes in cattle versus bison

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis is a primary agent of mastitis, pneumonia and arthritis in cattle and is the bacterium isolated most frequently from the polymicrobial syndrome known as bovine respiratory disease complex (BRDC). Recently, M. bovis has emerged as a significant health problem in bison, causing necro...

  11. The Phosphoproteome of the Minimal Bacterium Mycoplasma pneumoniae

    PubMed Central

    Schmidl, Sebastian R.; Gronau, Katrin; Pietack, Nico; Hecker, Michael; Becher, Dörte; Stülke, Jörg

    2010-01-01

    Mycoplasma pneumoniae belongs to the Mollicutes, the group of organisms with the smallest genomes that are capable of host-independent life. These bacteria show little regulation in gene expression, suggesting an important role for the control of protein activities. We have studied protein phosphorylation in M. pneumoniae to identify phosphorylated proteins. Two-dimensional gel electrophoresis and mass spectrometry allowed the detection of 63 phosphorylated proteins, many of them enzymes of central carbon metabolism and proteins related to host cell adhesion. We identified 16 phosphorylation sites, among them 8 serine and 8 threonine residues, respectively. A phosphoproteome analysis with mutants affected in the two annotated protein kinase genes or in the single known protein phosphatase gene suggested that only one protein (HPr) is phosphorylated by the HPr kinase, HPrK, whereas four adhesion-related or surface proteins were targets of the protein kinase C, PrkC. A comparison with the phosphoproteomes of other bacteria revealed that protein phosphorylation is evolutionarily only poorly conserved. Only one single protein with an identified phosphorylation site, a phosphosugar mutase (ManB in M. pneumoniae), is phosphorylated on a conserved serine residue in all studied organisms from archaea and bacteria to man. We demonstrate that this protein undergoes autophosphorylation. This explains the strong conservation of this phosphorylation event. For most other proteins, even if they are phosphorylated in different species, the actual phosphorylation sites are different. This suggests that protein phosphorylation is a form of adaptation of the bacteria to the specific needs of their particular ecological niche. PMID:20097688

  12. Three-dimensional morphology, ultrastructure, and replication of Mycoplasma felis.

    PubMed

    Boatman, E S; Kenny, G E

    1970-01-01

    The morphology and replication of Mycoplasma felis in relation to growth phase in culture were studied by electron microscopy. The organisms showed 1.0 to 1.45-hr doubling times with typical bacterial-type growth curves when grown in dialysate broth supplemented with horse serum. Organisms were fixed for electron microscopy by using Veronal acetate-buffered 0.8% OsO(4) (pH 6.1) in 20% sucrose. The morphology of exponential-phase organisms differed markedly from that of stationary or death-phase organisms, which were essentially large round forms with either dispersed or abnormally aggregated cytoplasm. Plasticine models prepared from serial sections of organisms in exponential phase showed the organisms to be either disc-shaped, triangular, horseshoe-shaped, or multilobular. A central "hole" was frequently present in these structures and could be visualized in the lobular forms as an interconnecting circular membrane. The inner surface of this membrane often showed contact with a small membranous body about 0.12 mum in diameter. The significance of this body is unknown. The morphology of the various shapes was confirmed by using the phosphotungstic acid and critical point methods. When the ratios of the various forms in exponential-phase cultures were determined, it was found that a replication sequence could be proposed which accounted for not only the volume increase required to accommodate deoxyribonucleic acid (DNA) replication but also the distribution of that DNA. Although it is likely that DNA replication in M. felis is a binary process, it appears that the mechanism for production of new cells need not be a binary process. PMID:5411752

  13. Evaluation of 10 serological assays for diagnosing Mycoplasma pneumoniae infection.

    PubMed

    Busson, Laurent; Van den Wijngaert, Sigi; Dahma, Hafid; Decolvenaer, Marc; Di Cesare, Lina; Martin, Agnes; Vasseur, Liesbet; Vandenberg, Olivier

    2013-06-01

    In this study, the performance of 10 serological assays for the diagnosis of Mycoplasma pneumoniae infection was evaluated. A total of 145 sera from 120 patients were tested. They were obtained from patients who were serologically positive for M. pneumoniae infection as well as from patients who were infected with micro-organisms that may cause interstitial pneumonia. The following assays were utilized: SeroMP IgM and IgG, SeroMP recombinant IgM, IgA and IgG, Liaison M. pneumoniae IgM and IgG and M. pneumoniae IgM, IgA and IgG ELISA Medac. The SeroMP Recombinant and Liaison assays both showed low IgM specificity, and crossreactivity was mainly observed in groups of patients with acute cytomegalovirus and Epstein-Barr virus infections. For IgA, the Medac assay was less specific than the SeroMP Recombinant assay. Discrepancies between the four tests were observed in IgG analyses, and due to the lack of a gold standard, 22 results were removed prior to determining the sensitivity and specificity. Therefore, the overall performance of IgG assays may be overstated; nevertheless, the SeroMP assay demonstrated a lack of sensitivity. The seroprevalence of IgG appears to be very low, raising concerns regarding whether the serological techniques can detect IgG levels over time. Serology remains a biological tool of choice for diagnosing M. pneumoniae infection, but improvement and standardization of the assays are needed, particularly for the determination of IgG. PMID:23537789

  14. Fatal Outcomes in Family Transmission of Mycoplasma pneumoniae

    PubMed Central

    Kannan, T. R.; Hardy, R. D.; Coalson, J. J.; Cavuoti, D. C.; Siegel, J. D.; Cagle, M.; Musatovova, O.; Herrera, C.

    2012-01-01

    Background.?Mycoplasma pneumoniae continues to be a significant cause of community-acquired pneumonia and, on rare occasions, manifests as fulminant disease that leads to mortality, even in healthy individuals. Methods.?We conducted a retrospective study on members of a family who were quarantined by the Centers for Disease Control and Prevention in 2002 for respiratory failure and death of a 15-year-old brother (sibling 1) and a 13-year-old sister (sibling 2). Collected airway, cerebrospinal fluid (CSF), and serum samples from both deceased siblings and serum samples from both parents and the remaining 3 ill siblings (sibling 3–5) were tested using a range of diagnostic assays. Autopsy lung tissue samples from sibling 2 were also assessed using immunohistochemical and immunoelectron microscopic methods. Results.?Autopsy evaluation of sibling 1 revealed cerebral edema consistent with hypoxic ischemic encepatholopathy and pulmonary findings of bronchiolitis obliterans with organizing pneumonia (BOOP). Postmortem lung examination of sibling 2 revealed lymphoplasmacytic bronchiolitis with intraluminal purulent exudate, BOOP, and pulmonary edema. Results of diagnostic assays implicated the household transmission of M. pneumoniae among all 5 siblings and both parents. Further analysis of lung tissue from sibling 2 demonstrated the presence of M. pneumoniae organisms and community-acquired respiratory distress syndrome toxin. M. pneumoniae was cultured directly from sibling 2 autopsy lung tissue. Conclusion.?Evidence is provided that M. pneumoniae was readily transmitted to all members of the household and that the resulting infections led to a spectrum of individual responses with variation in disease progression, including lymphoplasmacytic bronchiolitis, BOOP, and death. PMID:22052890

  15. A Trigger Enzyme in Mycoplasma pneumoniae: Impact of the Glycerophosphodiesterase GlpQ on Virulence and Gene Expression

    PubMed Central

    Schmidl, Sebastian R.; Otto, Andreas; Lluch-Senar, Maria; Piñol, Jaume; Busse, Julia; Becher, Dörte; Stülke, Jörg

    2011-01-01

    Mycoplasma pneumoniae is a causative agent of atypical pneumonia. The formation of hydrogen peroxide, a product of glycerol metabolism, is essential for host cell cytotoxicity. Phosphatidylcholine is the major carbon source available on lung epithelia, and its utilization requires the cleavage of deacylated phospholipids to glycerol-3-phosphate and choline. M. pneumoniae possesses two potential glycerophosphodiesterases, MPN420 (GlpQ) and MPN566. In this work, the function of these proteins was analyzed by biochemical, genetic, and physiological studies. The results indicate that only GlpQ is an active glycerophosphodiesterase. MPN566 has no enzymatic activity as glycerophosphodiesterase and the inactivation of the gene did not result in any detectable phenotype. Inactivation of the glpQ gene resulted in reduced growth in medium with glucose as the carbon source, in loss of hydrogen peroxide production when phosphatidylcholine was present, and in a complete loss of cytotoxicity towards HeLa cells. All these phenotypes were reverted upon complementation of the mutant. Moreover, the glpQ mutant strain exhibited a reduced gliding velocity. A comparison of the proteomes of the wild type strain and the glpQ mutant revealed that this enzyme is also implicated in the control of gene expression. Several proteins were present in higher or lower amounts in the mutant. This apparent regulation by GlpQ is exerted at the level of transcription as determined by mRNA slot blot analyses. All genes subject to GlpQ-dependent control have a conserved potential cis-acting element upstream of the coding region. This element overlaps the promoter in the case of the genes that are repressed in a GlpQ-dependent manner and it is located upstream of the promoter for GlpQ-activated genes. We may suggest that GlpQ acts as a trigger enzyme that measures the availability of its product glycerol-3-phosphate and uses this information to differentially control gene expression. PMID:21966272

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  17. 9 CFR 147.6 - Procedure for determining the status of flocks reacting to tests for Mycoplasma gallisepticum...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Section 147.6 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE LIVESTOCK...Mycoplasma may be found in Isolation and Identification of Avian Pathogens, published by the American Association of Avian...

  18. Rapid detection of contagious caprine pleuropneumonia using a Mycoplasma capricolum subsp. capripneumoniae capsular polysaccharide-specific antigen detection latex agglutination test.

    PubMed

    March, J B; Gammack, C; Nicholas, R

    2000-11-01

    Latex microspheres (diameter, 8 microm) were coated with anti-Mycoplasma capricolum subsp. capripneumoniae polyclonal immunoglobulin G (IgG) antiserum (anti-F38 biotype). The coated microspheres, when used in a latex agglutination test (LAT), detected M. capricolum subsp. capripneumoniae antigen in the serum of goats with contagious caprine pleuropneumoniae (CCPP). Beads also agglutinated strongly in the presence of purified M. capricolum subsp. capripneumoniae capsular polysaccharide (CPS). Preabsorption of CPS-specific antibodies prior to coating of the beads removed agglutinating activity in the presence of M. capricolum subsp. capripneumoniae, strongly suggesting that CPS is the likely soluble antigen recognized by the test. In addition, the specificity of the LAT exactly mirrored that of an M. capricolum subsp. capripneumoniae CPS-specific monoclonal antibody (WM25): of the 8 other mycoplasma species tested, agglutination was observed only with bovine serogroup 7. The LAT detected all 11 strains of M. capricolum subsp. capripneumoniae examined in this study, with a sensitivity level of 2 ng of CPS, or the equivalent of 1.7 x 10(4) CFU, in a reaction volume of 0.03 ml of serum. With field sera from goats with CCPP, the results of the LAT exhibited a 67% correlation with the results of the currently used complement fixation test (CFT), with the main discrepancy in diagnosis resulting from the increased sensitivity of the LAT compared to that of CFT. This antigen-detection LAT should prove particularly useful in identifying animals in the earliest stages of CCPP and combines sensitivity and low cost with ease of application in the field, without the need for any specialist training or equipment. PMID:11060083

  19. Rapid Detection of Contagious Caprine Pleuropneumonia Using a Mycoplasma capricolum subsp. capripneumoniae Capsular Polysaccharide-Specific Antigen Detection Latex Agglutination Test

    PubMed Central

    March, J. B.; Gammack, C.; Nicholas, R.

    2000-01-01

    Latex microspheres (diameter, 8 ?m) were coated with anti-Mycoplasma capricolum subsp. capripneumoniae polyclonal immunoglobulin G (IgG) antiserum (anti-F38 biotype). The coated microspheres, when used in a latex agglutination test (LAT), detected M. capricolum subsp. capripneumoniae antigen in the serum of goats with contagious caprine pleuropneumoniae (CCPP). Beads also agglutinated strongly in the presence of purified M. capricolum subsp. capripneumoniae capsular polysaccharide (CPS). Preabsorption of CPS-specific antibodies prior to coating of the beads removed agglutinating activity in the presence of M. capricolum subsp. capripneumoniae, strongly suggesting that CPS is the likely soluble antigen recognized by the test. In addition, the specificity of the LAT exactly mirrored that of an M. capricolum subsp. capripneumoniae CPS-specific monoclonal antibody (WM25): of the 8 other mycoplasma species tested, agglutination was observed only with bovine serogroup 7. The LAT detected all 11 strains of M. capricolum subsp. capripneumoniae examined in this study, with a sensitivity level of 2 ng of CPS, or the equivalent of 1.7 × 104 CFU, in a reaction volume of 0.03 ml of serum. With field sera from goats with CCPP, the results of the LAT exhibited a 67% correlation with the results of the currently used complement fixation test (CFT), with the main discrepancy in diagnosis resulting from the increased sensitivity of the LAT compared to that of CFT. This antigen-detection LAT should prove particularly useful in identifying animals in the earliest stages of CCPP and combines sensitivity and low cost with ease of application in the field, without the need for any specialist training or equipment. PMID:11060083

  20. Assessing the prevalence of mycoplasma contamination in cell culture via a survey of NCBI's RNA-seq archive

    PubMed Central

    Olarerin-George, Anthony O.; Hogenesch, John B.

    2015-01-01

    Mycoplasmas are notorious contaminants of cell culture and can have profound effects on host cell biology by depriving cells of nutrients and inducing global changes in gene expression. Over the last two decades, sentinel testing has revealed wide-ranging contamination rates in mammalian culture. To obtain an unbiased assessment from hundreds of labs, we analyzed sequence data from 9395 rodent and primate samples from 884 series in the NCBI Sequence Read Archive. We found 11% of these series were contaminated (defined as ?100 reads/million mapping to mycoplasma in one or more samples). Ninety percent of mycoplasma-mapped reads aligned to ribosomal RNA. This was unexpected given 37% of contaminated series used poly(A)-selection for mRNA enrichment. Lastly, we examined the relationship between mycoplasma contamination and host gene expression in a single cell RNA-seq dataset and found 61 host genes (P < 0.001) were significantly associated with mycoplasma-mapped read counts. In all, this study suggests mycoplasma contamination is still prevalent today and poses substantial risk to research quality. PMID:25712092

  1. Assessing the prevalence of mycoplasma contamination in cell culture via a survey of NCBI's RNA-seq archive.

    PubMed

    Olarerin-George, Anthony O; Hogenesch, John B

    2015-03-11

    Mycoplasmas are notorious contaminants of cell culture and can have profound effects on host cell biology by depriving cells of nutrients and inducing global changes in gene expression. Over the last two decades, sentinel testing has revealed wide-ranging contamination rates in mammalian culture. To obtain an unbiased assessment from hundreds of labs, we analyzed sequence data from 9395 rodent and primate samples from 884 series in the NCBI Sequence Read Archive. We found 11% of these series were contaminated (defined as ?100 reads/million mapping to mycoplasma in one or more samples). Ninety percent of mycoplasma-mapped reads aligned to ribosomal RNA. This was unexpected given 37% of contaminated series used poly(A)-selection for mRNA enrichment. Lastly, we examined the relationship between mycoplasma contamination and host gene expression in a single cell RNA-seq dataset and found 61 host genes (P < 0.001) were significantly associated with mycoplasma-mapped read counts. In all, this study suggests mycoplasma contamination is still prevalent today and poses substantial risk to research quality. PMID:25712092

  2. The Genome of the Obligate Intracellular Parasite Trachipleistophora hominis: New Insights into Microsporidian Genome Dynamics and Reductive Evolution

    PubMed Central

    Heinz, Eva; Williams, Tom A.; Nakjang, Sirintra; Noël, Christophe J.; Swan, Daniel C.; Goldberg, Alina V.; Harris, Simon R.; Weinmaier, Thomas; Markert, Stephanie; Becher, Dörte; Bernhardt, Jörg; Dagan, Tal; Hacker, Christian; Lucocq, John M.; Schweder, Thomas; Rattei, Thomas; Hall, Neil; Hirt, Robert P.; Embley, T. Martin

    2012-01-01

    The dynamics of reductive genome evolution for eukaryotes living inside other eukaryotic cells are poorly understood compared to well-studied model systems involving obligate intracellular bacteria. Here we present 8.5 Mb of sequence from the genome of the microsporidian Trachipleistophora hominis, isolated from an HIV/AIDS patient, which is an outgroup to the smaller compacted-genome species that primarily inform ideas of evolutionary mode for these enormously successful obligate intracellular parasites. Our data provide detailed information on the gene content, genome architecture and intergenic regions of a larger microsporidian genome, while comparative analyses allowed us to infer genomic features and metabolism of the common ancestor of the species investigated. Gene length reduction and massive loss of metabolic capacity in the common ancestor was accompanied by the evolution of novel microsporidian-specific protein families, whose conservation among microsporidians, against a background of reductive evolution, suggests they may have important functions in their parasitic lifestyle. The ancestor had already lost many metabolic pathways but retained glycolysis and the pentose phosphate pathway to provide cytosolic ATP and reduced coenzymes, and it had a minimal mitochondrion (mitosome) making Fe-S clusters but not ATP. It possessed bacterial-like nucleotide transport proteins as a key innovation for stealing host-generated ATP, the machinery for RNAi, key elements of the early secretory pathway, canonical eukaryotic as well as microsporidian-specific regulatory elements, a diversity of repetitive and transposable elements, and relatively low average gene density. Microsporidian genome evolution thus appears to have proceeded in at least two major steps: an ancestral remodelling of the proteome upon transition to intracellular parasitism that involved reduction but also selective expansion, followed by a secondary compaction of genome architecture in some, but not all, lineages. PMID:23133373

  3. Survival capacity of Mycoplasma agalactiae and Mycoplasma mycoides subsp capri in the diluted semen of goat bucks and their effects on sperm quality.

    PubMed

    Gómez-Martín, A; Uc, N; Vieira, L A; Gadea, J; Cadenas, J; Sánchez, A; De la Fe, C

    2015-03-15

    This study examines the viability of Mycoplasma agalactiae (Ma) and Mycoplasma mycoides subsp capri (Mmc) during 150 minutes of incubation at 37 °C in contaminated diluted semen (DS) doses. The effects of the presence of both microorganisms on sperm viability, motility, and morphology were also examined. In a second experiment, the viability of Ma and its effects on sperm viability were determined in ejaculate samples and skimmed milk semen extender samples. Ma and Mmc were able to survive in DS at concentrations considered infectious, and no significant differences in mean concentrations were detected (7.1 log colony-forming units [CFU]/mL). However, initial concentration of Ma declined (P < 0.05) from 7.5 to 6.9 log CFU/mL and Mmc declined (P < 0.05) from 7.7 to 7.1 log CFU/mL after incubation. Conversely, ejaculate concentrations of Ma increased significantly (from 7.1 to 7.4 log CFU/mL, P < 0.05). These observations suggest that the natural breeding medium is more suitable for Ma than the medium used for artificial insemination (AI). The presence of Mmc slightly reduced sperm viability in the DS (from 21.7% to 16.6%, P < 0.05). The absence of major effects on sperm quality could lead to the unnoticed use of semen contaminated with Ma and Mmc for AI. As both bacteria were able to survive the conditions of ejaculates and semen doses, these findings suggest a risk of venereal transmission of contagious agalactia and support the use of mycoplasma-free semen samples for (AI). PMID:25543157

  4. Infection dynamics of porcine reproductive and respiratory syndrome virus in a continuous-flow population of pigs also infected with Mycoplasma hyopneumoniae.

    PubMed

    Fano, E; Pijoan, C; Dee, S

    2007-10-13

    Twenty-eight 10-week-old pigs were inoculated intratracheally with 1 x 10(5) colour-changing units/ml Mycoplasma hyopneumoniae strain 232, and another 32 pigs were not inoculated but were divided into 12 direct-contact pigs and 20 indirect-contact pigs. Thirty-five days later, the inoculated pigs were inoculated intranasally with 1 x 10(2.4) tcid50 of porcine reproductive and respiratory syndrome virus (PRRSV) strain mn 30-100. Viraemia, seroconversion and the transmission of PRRSV in the M hyopneumoniae-infected pigs were then assessed for four months. Three groups of 10 age-matched gilts were introduced as sentinels into the experimental barn on days 28, 56 and 84 after the PRRSV infection. The persistence of PRRSV was evaluated in both the experimentally and naturally infected pigs, which were slaughtered 120, 135 and 150 days after the infection. The period of viraemia and the extent of seroconversion were similar to those observed in studies of pigs infected only with PRRSV, suggesting that under the conditions of the study M hyopneumoniae did not affect these features of the disease. A delayed pattern in the seroconversion and proportion of pcr-positive pigs was observed in the direct and indirect contact groups, and the persistence of PRRSV in tissues was confirmed by pcr at 120 and 150 days after infection only in the directly inoculated pigs and not in the direct- or indirect-contact groups of pigs. PMID:17938409

  5. Development of a selective polymerase chain reaction assay for the detection of Mycoplasma mycoides subsp. Mycoides S.C. (contagious bovine pleuropneumonia agent).

    PubMed

    Dedieu, L; Mady, V; Lefevre, P C

    1994-12-01

    A new selective assay for the detection of Mycoplasma mycoides subsp. mycoides SC (MmmSC) via the polymerase chain reaction (PCR) has been developed. This test used two PCR assays: a control-PCR (MYC-PCR) identifying the pathogen as a member of the mycoides cluster and the MSC-PCR which is specific for MmmSC. The MYC primers targeted a DNA sequence of about 460 bp from all the 59 mycoides cluster-strains tested. No amplification occurred with bovine genomic DNA or with the 11 other bacterial species assayed. The MSC primers selectively amplified a 275 bp sequence from the 27 MmmSC strains tested, with three specific internal restriction sites allowing confirmation of the identification. The sensitivity assessed by direct agarose gel analysis for both PCR assays was 100 CFU. The sensitivity of the MSC-PCR was increased to 1 CFU by a dot-blot hybridization step using, as a probe, the entire 275 bp sequence digoxigenin-labeled by PCR. These two PCR assays were successfully used to detect MmmSC in pleural fluids from naturally-infected cattle. We conclude that these two PCR assays may be valuable tools for the diagnosis of contagious bovine pleuropneumonia. PMID:9133058

  6. Molecular analysis of the P97 cilium adhesin operon of Mycoplasma hyopneumoniae

    Microsoft Academic Search

    Tsungda Hsu; F. Chris Minion

    1998-01-01

    Mycoplasma hyopneumoniae causes an economically significant respiratory disease of swine called Enzootic Pneumonia. The disease process is initiated by adherence of M. hyopneumoniae to the cilia of swine respiratory epithelium through an interaction involving P97, a surface-associated protein, and cilia-specific receptors. Binding specificity is associated with a repeat region located near the C-terminus of the P97 protein. Further analysis of

  7. Septic shock, necrotizing pneumonitis, and meningoencephalitis caused by Mycoplasma pneumoniae in a child: a case report.

    PubMed

    Barreira, Eliane R; Souza, Daniela C; Góes, Patricia F; Bousso, Albert

    2009-04-01

    Mycoplasma pneumoniae is an important causative agent of respiratory infection in childhood. Although the infection caused by M. pneumoniae is classically described as benign, severe and life-threatening pulmonary and extrapulmonary complications can occur. This study describes the first case of septic shock related to M. pneumoniae in a child with necrotizing pneumonitis, severe encephalitis, and multiple organs involvement, with a favorable outcome after lobectomy and systemic corticosteroids. PMID:19023109

  8. Acute respiratory infection due to Mycoplasma pneumoniae : current status of diagnostic methods

    Microsoft Academic Search

    K. Loens; H. Goossens; M. Ieven

    2010-01-01

    Because of the absence of well-standardized both in-house and FDA-approved commercially available diagnostic tests, the reliable\\u000a diagnosis of respiratory infection due to Mycoplasma pneumoniae remains difficult. In addition, no formal external quality assessment schemes which would allow to conclude about the performance\\u000a of M. pneumoniae diagnostic tests exist. In this review, the current state of knowledge of M. pneumoniae-associated respiratory

  9. Mycoplasma-like bodies in Solanum laciniatum plants infected with potato witches’ broom disease

    Microsoft Academic Search

    Marie Ulrychová; M. Jokeš

    1977-01-01

    Mycoplasma-like organisms (MLO) spread from the infectious grafts intoSolanum laciniatum\\u000a Ait. stock plants relatively slowly. MLO were present in all sprouts ofS. laciniatum four weeks after grafting, but the infected plants remained under glasshouse conditions mostly symptomless and flowered normally\\u000a and formed fruits like healthy plants. The growth of plants with infectious tomato grafts was identical with the controls\\u000a but

  10. Evidence for Type III Restriction and Modification Systems in Mycoplasma pulmonis?

    PubMed Central

    Dybvig, Kevin; Cao, Z.; French, C. Todd; Yu, Huilan

    2007-01-01

    Mycoplasma pulmonis possesses a cassette of genes that are predicted to code for type III restriction and modification (R-M) enzymes. Transposon disruption of a gene predicted to code for the endonuclease subunit of the enzyme resulted in loss of R-M activity. Genomic data indicate that the cassette was acquired by horizontal gene transfer and possibly located on a mobile element. PMID:17209015

  11. Evaluation of Five Real-Time PCR Assays for Detection of Mycoplasma pneumoniae

    PubMed Central

    Jacobs, Enno

    2014-01-01

    Four commercial real-time PCR assays to detect Mycoplasma pneumoniae were tested, and the results were compared with the results for an in-house approach. Despite differences of crossing threshold values of up to 4, assays were able to detect at least 20 CFU/5 ?l (52 fg DNA/5 ?l) of sample with the Diagenode kit showing the best clinical sensitivity. PMID:25210063

  12. Mycoplasma pneumoniae : Innocent Bystander or a True Cause of Central Nervous System Disease?

    Microsoft Academic Search

    Ari Bitnun; Susan E. Richardson

    2010-01-01

    The consistency with which Mycoplasma pneumoniae has been implicated as a cause of encephalitis, and the increased incidence of central nervous system (CNS) disease observed\\u000a during M. pneumoniae respiratory outbreaks, support the role of M. pneumoniae as a CNS pathogen. Three pathophysiologic mechanisms have been proposed: direct infection, autoimmunity, and vascular occlusion.\\u000a Recent evidence demonstrating the organism’s ability to survive

  13. Mycoplasma pneumonia associated with rhabdomyolysis and the Guillain-Barre syndrome.

    PubMed

    Gupta, R; Gupta, A; Goyal, V; Guleria, R; Kumar, A

    2005-01-01

    A 25-year-old housewife who presented with Mycoplasma pneumonia who developed acute respiratory distress syndrome (ARDS) and required assisted ventilation. During her hospital stay, she developed acute renal failure because of rhabdomyolysis and was put on haemodialysis. She also had difficulty in weaning from ventilator because of acute motor-sensory axonal neuropathy (AMSAN) variant of the Guillain-Barre syndrome. The patient was treated with antibiotics and corticosteroids. The patient recovered from both the complications gradually. PMID:16255404

  14. Reactive Species Mediate Inhibition of Alveolar Type II Sodium Transport during Mycoplasma Infection

    PubMed Central

    Hickman-Davis, Judy M.; McNicholas-Bevensee, Carmel; Davis, Ian C.; Ma, He-Ping; Davis, Glenda C.; Bosworth, Charles A.; Matalon, Sadis

    2006-01-01

    Rationale: Mycoplasma pneumoniae is a significant cause of pneumonia in humans. Objectives: To determine the impact of mycoplasma infection and the host inflammatory response on alveolar type II (ATII) cell ion transport in vivo and in vitro. Methods: Mice were infected with M. pulmonis for measurements of alveolar fluid clearance (AFC) in vivo and isolation of ATII cells. ATII cells were infected in vivo for determination of epithelial Na+ channel (ENaC) total and cell surface protein levels by biotinylation and Western blot and in vitro for whole cell patch clamp recording and measurement of nitric oxide (NO) production by chemiluminescence. Results: Mycoplasma infection significantly inhibited AFC at 24 h and total and amiloride-sensitive AFC by 48 h postinfection (pi). In contrast, infected myeloperoxidase-deficient mice had similar basal and amiloride-sensitive AFC values to uninfected control mice at 48 h pi. Addition of forskolin restored total and amiloride-sensitive AFC to control values at 48 h pi. ATII cells isolated from infected mice demonstrated normal ?, ?, and ? ENaC total protein levels; however, infected whole-lung cell-surface levels of ? ENaC were significantly decreased. Patch-clamp recordings demonstrated a significant decrease in total and amiloride-sensitive Na+ currents at 24 h pi. ATII cells demonstrated a significant increase in the production of NO at 24 h pi and inhibition of NO by ATII cells before infection reversed the decrease in total Na+ currents. Conclusions: These data indicate that mycoplasma infection results in decreased AFC and functional ENaC via the production of reactive oxygen nitrogen intermediates. PMID:16254273

  15. Mutant Analysis Reveals a Specific Requirement for Protein P30 in Mycoplasma pneumoniae Gliding Motility

    Microsoft Academic Search

    Benjamin M. Hasselbring; Jarrat L. Jordan; Duncan C. Krause

    2005-01-01

    The cell-wall-less prokaryote Mycoplasma pneumoniae, long considered among the smallest and simplest cells capable of self-replication, has a distinct cellular polarity characterized by the presence of a differentiated terminal organelle which functions in adherence to human respiratory epithelium, gliding motility, and cell division. Characterization of hemadsorption (HA)-negative mutants has resulted in identification of several terminal organelle proteins, including P30, the

  16. Mycoplasma-Like organisms associated with stunting of Gypsophila paniculata L

    Microsoft Academic Search

    Marie Ulrychová; Eva Petr?; M. Jokeš; Blanka Joštová

    1983-01-01

    A new undescribed yellows-type disease ofGypsophila paniculata L. was found. Since a stunted growth is the predominant symptom the disease has been termed stunting. Electron micrographs\\u000a of ultrathin sections revealed numerous mycoplasma-like organisms in diseased phloem tissues. Attempts to maintain the diseasedGypsophila plants for further investigation of this disease failed for the present in spite of all applied modifications in

  17. Mycoplasma genitalium is not associated with adverse outcomes of pregnancy in Guinea-Bissau

    Microsoft Academic Search

    A-C Labbe?; E Frost; S Deslandes; A P Mendonc?a; A C Alves; J Pe?pin

    2002-01-01

    Objective: To evaluate the impact of Mycoplasma genitalium on the outcome of pregnancy.Methods: Cervical samples from women who had previously participated in a case-control study (designed to assess the impact of syphilis and HIV-2 on the outcome of pregnancy in Guinea-Bissau) were processed using a PCR assay to detect the presence of M genitalium. Controls were women who had delivered

  18. Drug Resistance–Associated Mutations in Mycoplasma genitalium in Female Sex Workers, Japan

    PubMed Central

    Yasuda, Mitsuru; Horie, Kengo; Seike, Kensaku; Kikuchi, Mina; Mizutani, Kohsuke; Tsuchiya, Tomohiro; Yokoi, Shigeaki; Nakano, Masahiro; Hoshina, Shinji

    2015-01-01

    Mycoplasma genitalium was detected in 21 (14.1%) of 149 vaginal swab samples and in 1 (0.7%) of 149 throat washing samples from female sex workers during 2013–2014 in Japan. Prevalences of M. genitalium with macrolide resistance–associated 23S rRNA mutations and fluoroquinolone resistance–associated parC alterations were 47.1% and 36.8%, respectively. PMID:25988775

  19. Scanning electron microscopy of Mycoplasma pneumoniae on the membrane of individual ciliated tracheal cells

    Microsoft Academic Search

    Michael G. Gabridge; Marlene J. Bright; Helen R. Richards

    1982-01-01

    Summary  Cell monolayer cultures were prepared from hamster tracheal explants by a collagenase exposure and subsequent incubation in\\u000a Waymouth’s MAB 87\\/3 medium. The epithelial outgrowth occurred on glass cover slips. Cilia on the monolayers continued to beat\\u000a normally after the “parent” explant was removed. Monolayer cultures infected withMycoplasma pneumoniae had significant amounts of attachment. A morphological analysis of the attachment was

  20. Loss of diversity within Mycoplasma bovis isolates collected in France from bovines with respiratory diseases over the last 35years.

    PubMed

    Becker, Claire A M; Thibault, François M; Arcangioli, Marie-Anne; Tardy, Florence

    2015-07-01

    Mycoplasma (M.) bovis has recently emerged as a major, worldwide etiological agent of bovine respiratory diseases leading to huge economic losses mainly due to high morbidity and mortality as well as poor growth rates. The spread of M. bovis infections between different animals, herds, regions or countries has been often reported to be connected to the movement of animals. However, despite recent considerable efforts, no universal subtyping method is yet available to trace M. bovis isolates circulation at an international scale. Moreover in France, the overall population diversity of M. bovis isolates has not been assessed since the early 1990s. This study was conducted to fill in these gaps. The genotypic diversity between sixty isolates collected in France over the last 35years was assessed using two molecular subtyping methods that addressed either the long-term epidemiological relationships (Multi Locus Sequence Typing, MLST) or the genetic microvariations (Multiple Locus VNTR Analysis, MLVA) between isolates. Phenotypic diversity was also analyzed by using Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) to compare the main protein patterns of isolates. All proposed subtyping approaches were optimized and led to the same pattern in the French M. bovis population that consisted of two clusters, the first one comprising isolates collected before 2000 and the second, those collected after 2000. Recent strains were further shown to be more homogeneous than older ones, which is consistent with the spread of a single clone throughout the country. Because this spread was concomitant with the emergence of multiresistant M. bovis isolates, several hypotheses are discussed to explain the homogeneity of M. bovis isolates in France, even though the M. bovis species is fully equipped to generate diversity. PMID:25913158

  1. Field evaluation of a quantitative polymerase chain reaction assay for Mycoplasma hyorhinis.

    PubMed

    Clavijo, Maria J; Oliveira, Simone; Zimmerman, Jeffrey; Rendahl, Aaron; Rovira, Albert

    2014-11-01

    Mycoplasma hyorhinis has emerged as an important cause of systemic disease in nursery pigs. However, this bacterium can also be found in the upper respiratory tract of healthy swine. The current study describes the development of a quantitative polymerase chain reaction assay for the detection of M. hyorhinis and the evaluation of the assay in both disease diagnosis and disease surveillance using a large number of field samples. The analytical sensitivity was estimated to be 12 genome equivalents/?l. The assay was highly specific, detecting all 25 M. hyorhinis isolates tested and none of the 19 nontarget species tested. Assay repeatability was evaluated by testing different matrices spiked with known amounts of M. hyorhinis. Overall, assessment of the repeatability of the assay showed suitable precision within and between runs for all matrices. The coefficient of variation ranged from 10% to 24%. Mycoplasma hyorhinis DNA was detected in 48% of samples (pericardium, pleura, joints, nasal cavity, and lungs) from pigs with systemic disease. Mycoplasma hyorhinis was detected in nasal (92%) and oropharyngeal swabs (66%), as well as in oral fluids (100%). Potential uses of this tool involve the characterization of the prevalence of this pathogen in swine herds as well as bacterial quantification to evaluate intervention efficacy. PMID:25319032

  2. Mycoplasma pneumoniae protein P30 is required for cytadherence and associated with proper cell development.

    PubMed

    Romero-Arroyo, C E; Jordan, J; Peacock, S J; Willby, M J; Farmer, M A; Krause, D C

    1999-02-01

    The attachment organelle of Mycoplasma pneumoniae is a polar, tapered cell extension containing an intracytoplasmic, electron-dense core. This terminal structure is the leading end in gliding motility, and its duplication is thought to precede cell division, raising the possibility that mutations affecting cytadherence also confer a defect in motility or cell development. Mycoplasma surface protein P30 is associated with the attachment organelle, and P30 mutants II-3 and II-7 do not cytadhere. In this study, the recombinant wild-type but not the mutant II-3 p30 allele restored cytadherence when transformed into P30 mutants by recombinant transposon delivery. The mutations associated with loss of P30 in mutant II-3 and reacquisition of P30 in cytadhering revertants thereof were identified by nucleotide sequencing of the p30 gene. Morphological abnormalities that included ovoid or multilobed cells having a poorly defined tip structure were associated with loss of P30. Digital image analysis confirmed quantitatively the morphological differences noted visually. Transformation of the P30 mutants with the wild-type p30 allele restored a normal morphology, as determined both visually and by digital image analysis, suggesting that P30 plays a role in mycoplasma cell development. Finally, the P30 mutants localized the adhesin protein P1 to the terminal organelle, indicating that P30 is not involved in P1 trafficking but may be required for its receptor-binding function. PMID:9973332

  3. Mycoplasma pneumoniae Protein P30 Is Required for Cytadherence and Associated with Proper Cell Development

    PubMed Central

    Romero-Arroyo, Cynthia E.; Jordan, Jarrat; Peacock, Susan J.; Willby, Melisa J.; Farmer, Mark A.; Krause, Duncan C.

    1999-01-01

    The attachment organelle of Mycoplasma pneumoniae is a polar, tapered cell extension containing an intracytoplasmic, electron-dense core. This terminal structure is the leading end in gliding motility, and its duplication is thought to precede cell division, raising the possibility that mutations affecting cytadherence also confer a defect in motility or cell development. Mycoplasma surface protein P30 is associated with the attachment organelle, and P30 mutants II-3 and II-7 do not cytadhere. In this study, the recombinant wild-type but not the mutant II-3 p30 allele restored cytadherence when transformed into P30 mutants by recombinant transposon delivery. The mutations associated with loss of P30 in mutant II-3 and reacquisition of P30 in cytadhering revertants thereof were identified by nucleotide sequencing of the p30 gene. Morphological abnormalities that included ovoid or multilobed cells having a poorly defined tip structure were associated with loss of P30. Digital image analysis confirmed quantitatively the morphological differences noted visually. Transformation of the P30 mutants with the wild-type p30 allele restored a normal morphology, as determined both visually and by digital image analysis, suggesting that P30 plays a role in mycoplasma cell development. Finally, the P30 mutants localized the adhesin protein P1 to the terminal organelle, indicating that P30 is not involved in P1 trafficking but may be required for its receptor-binding function. PMID:9973332

  4. Quantitative real-time polymerase chain reaction for detecting Mycoplasma hyosynoviae and Mycoplasma hyorhinis in pen-based oral, tonsillar, and nasal fluids

    PubMed Central

    Bower, Leslie; Erickson, Barbara Z.; Wang, Chong; Raymond, Matthew; Strait, Erin L.

    2015-01-01

    Mycoplasma (M.) hyorhinis and M. hyosynoviae are pathogens known to cause disease in pigs post-weaning. Due to their fastidious nature, there is increased need for culture-independent diagnostic platforms to detect these microorganisms. Therefore, this study was performed to develop and optimize quantitative real-time PCR (qPCR) assays to rapidly detect M. hyorhinis and M. hyosynoviae in pen-based oral fluids as well as nasal and tonsillar fluids as proxies for samples used in swine herd surveillance. Two methods of genomic DNA extraction, automated versus manual, were used to compare diagnostic test performance. A wean-to-finish longitudinal study was also carried out to demonstrate the reproducibility of using pen-based oral fluids. Overall, pen-based oral and tonsillar fluids were more likely to be positive for both types of bacteria whereas only M. hyorhinis was detected in nasal fluids. DNA extraction protocols were shown to significantly influence test result. Although the initial detection time somewhat differed, both organisms were repeatedly detected in the longitudinal study. Overall, this study evaluated two qPCR methods for rapid and specific detection of either mycoplasma. Results from the present investigation can serve as a foundation for future studies to determine the prevalence of the two microorganisms, environmental load, and effectiveness of veterinary interventions for infection control. PMID:25643803

  5. A survey of Mycoplasma gallisepticum and Mycoplasma synovaie with avian influenza H9 subtype in meat-type chicken in Jordan between 2011-2015.

    PubMed

    Roussan, Dergham Ahmad; Khawaldeh, Ghassan; Shaheen, Ibrahim Ali

    2015-07-01

    Commercial chickens in Jordan suffer from respiratory disease of undetermined etiology. This study was designed to document the involvement of avian influenza virus (AIV) H9 subtype, Mycoplasma gallisepticum (MG), and Mycoplasma synoviae (MS) in this respiratory disease. In this study, trachea swabs from 350 commercial broiler chicken flocks that suffered from respiratory disease were tested for AIV H9 subtype by using reverse transcription (RT)-PCR and for MG and MS by using PCR. PCR and RT-PCR results showed that 23.7, 8.9, and 6.6% of these flocks were infected with AIV H9 subtype, MS, and MG, respectively, whereas 12.9 and 5.7% of these flocks were infected with both AIV H9 subtype and MS and AIV H9 subtype and MS, respectively. Furthermore, 42.3% of these flocks were negative for the above mentioned respiratory diseases. Further epidemiological studies are recommended to determine risk factors and evaluate the economic consequences of AIV H9 subtype, MG, and MS infections in the region. Furthermore, studies are required to isolate AIV H9 subtype, MG, and MS and develop vaccines against the local field isolates. PMID:25971950

  6. Quantitative real-time polymerase chain reaction for detecting Mycoplasma hyosynoviae and Mycoplasma hyorhinis in pen-based oral, tonsillar, and nasal fluids.

    PubMed

    Gomes Neto, João Carlos; Bower, Leslie; Erickson, Barbara Z; Wang, Chong; Raymond, Matthew; Strait, Erin L

    2015-06-01

    Mycoplasma (M.) hyorhinis and M. hyosynoviae are pathogens known to cause disease in pigs post-weaning. Due to their fastidious nature, there is increased need for culture-independent diagnostic platforms to detect these microorganisms. Therefore, this study was performed to develop and optimize quantitative real-time PCR (qPCR) assays to rapidly detect M. hyorhinis and M. hyosynoviae in pen-based oral fluids as well as nasal and tonsillar fluids as proxies for samples used in swine herd surveillance. Two methods of genomic DNA extraction, automated versus manual, were used to compare diagnostic test performance. A wean-to-finish longitudinal study was also carried out to demonstrate the reproducibility of using pen-based oral fluids. Overall, pen-based oral and tonsillar fluids were more likely to be positive for both types of bacteria whereas only M. hyorhinis was detected in nasal fluids. DNA extraction protocols were shown to significantly influence test result. Although the initial detection time somewhat differed, both organisms were repeatedly detected in the longitudinal study. Overall, this study evaluated two qPCR methods for rapid and specific detection of either mycoplasma. Results from the present investigation can serve as a foundation for future studies to determine the prevalence of the two microorganisms, environmental load, and effectiveness of veterinary interventions for infection control. PMID:25643803

  7. Comparative assessment of two commonly used commercial ELISA tests for the serological diagnosis of contagious agalactia of small ruminants caused by Mycoplasma agalactiae

    PubMed Central

    2012-01-01

    Background Contagious agalactia (CA) of sheep and goats caused by Mycoplasma agalactiae is a widely occurring economically important disease that is difficult to control. The ELISA is commonly used for the serological detection of CA but it has some limitations and the performance of the available tests have not been properly evaluated. Two commercial ELISA kits are widely used, one involving a fusion protein as target antigen and the other a total antigen. The objectives were to compare these tests by evaluating: i. Their diagnostic sensitivity and specificity, the relevance of the recommended cut-off points, the correlation between the two tests, and, the correlation between serology data and the milk shedding of M. agalatiae; ii. The influence of extrinsic factors such as the targeted animal species, geographical origin of the samples, intra-specific variability of M. agalactiae and concurrent mycoplasma infections. A sample of 5900 animals from 211 farms with continuous CA monitoring for 20?years and no prior vaccination history was used. The infection status was known from prior bacteriological, epidemiological and serological monitoring with a complementary immunoblotting test. Results The average diagnostic sensitivity was 56% [51.8–59.8] for the fusion protein ELISA and 84% [81.3–87.2] for the total antigen ELISA, with noteworthy flock-related variations. The average diagnostic specificity for the fusion protein ELISA was 100% [99.9–100], and for the total antigen ELISA differed significantly between goats and sheep: 99.3% [97.4–99.9] and 95.7% [93.8–97.2] respectively. Experimental inoculations with different M. agalactiae strains revealed that the ELISA kits poorly detected the antibody response to certain strains. Furthermore, test performances varied according to the host species or geographical origin of the samples. Finally, the correlation between milk shedding of M. agalactiae and the presence of detectable antibodies in the blood was poor. Conclusions These serological tests are not interchangeable. The choice of a test will depend on the objectives (early detection of infection or disease control program), on the prevalence of infection and the control protocol used. Given the variety of factors that may influence performance, a preliminary assessment of the test in a given situation is recommended prior to widespread use. PMID:22776779

  8. Naturally occurring aminoacyl-tRNA synthetases editing-domain mutations that cause mistranslation in Mycoplasma parasites

    PubMed Central

    Li, Li; Boniecki, Michal T.; Jaffe, Jacob D.; Imai, Brian S.; Yau, Peter M.; Luthey-Schulten, Zaida A.; Martinis, Susan A.

    2011-01-01

    Mycoplasma parasites escape host immune responses via mechanisms that depend on remarkable phenotypic plasticity. Identification of these mechanisms is of great current interest. The aminoacyl-tRNA synthetases (AARSs) attach amino acids to their cognate tRNAs, but occasionally make errors that substitute closely similar amino acids. AARS editing pathways clear errors to avoid mistranslation during protein synthesis. We show here that AARSs in Mycoplasma parasites have point mutations and deletions in their respective editing domains. The deleterious effect on editing was confirmed with a specific example studied in vitro. In vivo mistranslation was determined by mass spectrometric analysis of proteins produced in the parasite. These mistranslations are uniform cases where the predicted closely similar amino acid replaced the correct one. Thus, natural AARS editing-domain mutations in Mycoplasma parasites cause mistranslation. We raise the possibility that these mutations evolved as a mechanism for antigen diversity to escape host defense systems. PMID:21606343

  9. Biofilm Formation and Antimicrobial Susceptibility of Staphylococcus epidermidis Strains from a Hospital Environment

    PubMed Central

    Wojtyczka, Robert D.; Orlewska, Kamila; K?pa, Ma?gorzata; Idzik, Danuta; Dziedzic, Arkadiusz; Mularz, Tomasz; Krawczyk, Micha?; Miklasi?ska, Maria; W?sik, Tomasz J.

    2014-01-01

    The hospital environment microflora comprise a wide variety of microorganisms which are more or less pathogenic and where staphylococci are one of the most common types. The aim of the presented study was to evaluate the prevalence of the biofilm forming coagulase-negative staphylococci (CoNS) in a hospital environment as a risk factor for nosocomial infections. Among 122 isolated and tested strains of CoNS the most frequent were: S. epidermidis—32 strains, S. haemolyticus—31 strains, S. capitis subsp. capitis—21 strains, S. hominis—11 strains, S. cohnii subsp. cohnii—nine strains. In case of CoNS, the main molecule responsible for intercellular adhesion is a polysaccharide intercellular adhesin (PIA), encoded on the ica gene operon. The analysis revealed the presence of the icaADBC operon genes in 46.88% of S. epidermidis isolates. IcaA and icaD were present in 34.38% and 28.13% of strains respectively while IcaC gene was present in 37.50% of strains. IcaB gene was found in 21.88% of S. epidermidis strains. In 15 (63%) strains all icaADBC operon genes were observed. The assessment of antibacterial drugs susceptibility demonstrated that analyzed CoNS strains were highly resistant to macrolides and lincosamides and more sensitive to rifampicin and linezolid. Our data indicates that the hospital environment can be colonized by biofilm forming coagulase-negative staphylococci and transmission of these strains can cause an increased risk of serious nosocomial infections. PMID:24776724

  10. Suppression-Subtractive Hybridization as a Strategy To Identify Taxon-Specific Sequences within the Mycoplasma mycoides Cluster: Design and Validation of an M. capricolum subsp. capricolum-Specific PCR Assay?

    PubMed Central

    Maigre, Laure; Citti, Christine; Marenda, Marc; Poumarat, François; Tardy, Florence

    2008-01-01

    The phylogenetically related Mycoplasma capricolum subsp. capricolum and M. mycoides subsp. mycoides biotype Large Colony are two small-ruminant pathogens involved in contagious agalactia. Their respective contributions to clinical outbreaks are not well documented, because they are difficult to differentiate with the current diagnostic techniques. In order to identify DNA sequences specific to one taxon or the other, a suppression-subtractive hybridization approach was developed. DNA fragments resulting from the reciprocal subtraction of the type strains were used as probes on a panel of M. capricolum subsp. capricolum and M. mycoides subsp. mycoides biotype Large Colony strains to assess their intrataxon specificity. Due to a high intrataxon polymorphism and important cross-reactions between taxa, a single DNA fragment was shown to be specific for M. capricolum subsp. capricolum and to be present in all M. capricolum subsp. capricolum field isolates tested in this study. A PCR assay targeting the corresponding gene (simpA51) was designed that resulted in a 560-bp amplification only in M. capricolum subsp. capricolum and in M. capricolum subsp. capripneumoniae (the etiological agent of contagious caprine pleuropneumonia). simpA51 was further improved to generate a multiplex PCR (multA51) that allows the differentiation of M. capricolum subsp. capripneumoniae from M. capricolum subsp. capricolum. Both the simpA51 and multA51 assays accurately identify M. capricolum subsp. capricolum among other mycoplasmas, including all members of the M. mycoides cluster. simpA51 and multA51 PCRs are proposed as sensitive and robust tools for the specific identification of M. capricolum subsp. capricolum and M. capricolum subsp. capripneumoniae. PMID:18234866

  11. Disruption of the pdhB pyruvate dehyrogenase gene affects colony morphology, in vitro growth and cell invasiveness of Mycoplasma agalactiae.

    PubMed

    Hegde, Shivanand; Rosengarten, Renate; Chopra-Dewasthaly, Rohini

    2015-01-01

    The utilization of available substrates, the metabolic potential and the growth rates of bacteria can play significant roles in their pathogenicity. This study concentrates on Mycoplasma agalactiae, which causes significant economic losses through its contribution to contagious agalactia in small ruminants by as yet unknown mechanisms. This lack of knowledge is primarily due to its fastidious growth requirements and the scarcity of genetic tools available for its manipulation and analysis. Transposon mutagenesis of M. agalactiae type strain PG2 resulted in several disruptions throughout the genome. A mutant defective in growth in vitro was found to have a transposon insertion in the pdhB gene, which encodes a component of the pyruvate dehydrogenase complex. This growth difference was quite significant during the actively dividing logarithmic phase but a gradual recovery was observed as the cells approached stationary phase. The mutant also exhibited a different and smaller colony morphology compared to the wild type strain PG2. For complementation, pdhAB was cloned downstream of a strong vpma promoter and upstream of a lacZ reporter gene in a newly constructed complementation vector. When transformed with this vector the pdhB mutant recovered its normal growth and colony morphology. Interestingly, the pdhB mutant also had significantly reduced invasiveness in HeLa cells, as revealed by double immunofluorescence staining. This deficiency was recovered in the complemented strain, which had invasiveness comparable to that of PG2. Taken together, these data indicate that pyruvate dehydrogenase might be an important player in infection with and colonization by M. agalactiae. PMID:25799063

  12. Disruption of the pdhB Pyruvate Dehydrogenase Gene Affects Colony Morphology, In Vitro Growth and Cell Invasiveness of Mycoplasma agalactiae

    PubMed Central

    Hegde, Shivanand; Rosengarten, Renate; Chopra-Dewasthaly, Rohini

    2015-01-01

    The utilization of available substrates, the metabolic potential and the growth rates of bacteria can play significant roles in their pathogenicity. This study concentrates on Mycoplasma agalactiae, which causes significant economic losses through its contribution to contagious agalactia in small ruminants by as yet unknown mechanisms. This lack of knowledge is primarily due to its fastidious growth requirements and the scarcity of genetic tools available for its manipulation and analysis. Transposon mutagenesis of M. agalactiae type strain PG2 resulted in several disruptions throughout the genome. A mutant defective in growth in vitro was found to have a transposon insertion in the pdhB gene, which encodes a component of the pyruvate dehydrogenase complex. This growth difference was quite significant during the actively dividing logarithmic phase but a gradual recovery was observed as the cells approached stationary phase. The mutant also exhibited a different and smaller colony morphology compared to the wild type strain PG2. For complementation, pdhAB was cloned downstream of a strong vpma promoter and upstream of a lacZ reporter gene in a newly constructed complementation vector. When transformed with this vector the pdhB mutant recovered its normal growth and colony morphology. Interestingly, the pdhB mutant also had significantly reduced invasiveness in HeLa cells, as revealed by double immunofluorescence staining. This deficiency was recovered in the complemented strain, which had invasiveness comparable to that of PG2. Taken together, these data indicate that pyruvate dehydrogenase might be an important player in infection with and colonization by M. agalactiae. PMID:25799063

  13. Survey of Toxoplasma gondii and Neospora caninum, haemotropic mycoplasmas and other arthropod-borne pathogens in cats from Albania

    PubMed Central

    2014-01-01

    Background Albania is a country on the western part of the Balkan Peninsula. The Mediterranean climate is favourable for the stable development of many arthropod species, which are incriminated as vectors for various agents. Recently, several papers have reported on epidemiological aspects of parasitic diseases including vector-borne disease agents of dogs with zoonotic characteristics in Albania. However, data on the epidemiology of feline parasitic and bacterial agents in Albania is scarce. Methods Serum and EDTA-blood samples collected from 146 domestic cats from Tirana during 2008 through 2010 were examined for exposure to Toxoplasma gondii, Neospora caninum, Leishmania infantum, and Anaplasma spp. with IFAT, for infection with L. infantum, A. phagocytophilum, Bartonella spp. and haemotropic mycoplasmas with conventional PCR and real-time PCR and for Dirofilaria immitis with antigen ELISA. Additionally blood smear microscopy was carried out for detection of blood-borne pathogens. Results Antibodies to T. gondii (titre ?1:100) were demonstrated in 91 cats (62.3%). Antibodies to N. caninum (titre ?1:100), L. infantum (titre ?1:64) and Anaplasma spp. (titre ?1:100) were found in the serum of 15 (10.3%), 1 (0.7%) or 3 (2.1%) cats, respectively. DNA of haemotropic mycoplasmas was detected in the blood of 45 cats (30.8%), namely Candidatus Mycoplasma haemominutum (21.9%), Mycoplasma haemofelis (10.3%), and Candidatus Mycoplasma turicensis (5.5%), with ten cats harbouring co-infections of two mycoplasmas each; blood from one cat was PCR positive for Bartonella henselae. No DNA of Leishmania spp. and A. phagocytophilum or circulating D. immitis antigen was detected in any cat sample. The overall prevalence of haemotropic mycoplasmas was significantly higher in male compared to female cats (40.6% vs. 24.1%, p?=?0.0444); and age was associated positively with the prevalence of antibodies to T. gondii (p?=?0.0008) and the percentage of haemotropic mycoplasma infection (p?=?0.0454). Conclusions With the broad screening panel including direct and indirect methods applied in the present study, a wide spectrum of exposure to or infection with parasitic or bacterial agents was detected. PMID:24517118

  14. [Intestinal parasitosis in Indians of the Mapuera community (Oriximiná, State of Pará, Brazil): high prevalence of Blastocystis hominis and finding of Cryptosporidium sp and Cyclospora cayetanensis].

    PubMed

    Borges, Jaila Dias; Alarcón, Ruth Semira Rodríguez; Neto, Vicente Amato; Gakiya, Erika

    2009-01-01

    Occurrences of intestinal parasitosis in Indians of the Mapuera community (Oriximiná, State of Pará, Brazil) were evaluated. Within the context of group assessment, this study makes a contribution towards adequate knowledge of this subject, which is significant from a medical-sanitary point of view. Parasitological examination of feces from 83 individuals, performed using four different methods, could be considered to have reasonable amplitude for establishing diagnoses. Protozoan cysts and helminth eggs of many types were found, even with significant percentages. The frequent presence of Blastocystis hominis (57.8%), along with findings of Cryptosporidium sp (3.6%) and Cyclospora cayetanensis (10.8%), deserved highlighting with specific comments. The findings show that these Indians live in an environment in which poor hygiene conditions prevail. In particular, these facilitate the dissemination of protozoa and helminths through contact with the soil or through intake of contaminated water and food. PMID:19684989

  15. In Vitro efficacy of antimicrobial extracts against the atypical ruminant pathogen Mycoplasma mycoides subsp. capri

    PubMed Central

    2012-01-01

    Background Mycoplasmosis is a common infection in human and veterinary medicine, and is associated with chronic inflammation and high morbidity. Mycoplasma species are often intrinsically resistant to many conventional antimicrobial therapies, and the resistance patterns of pathogenic mycoplasmas to commonly used medicinal (antimicrobial) plant extracts are currently unknown. Methods Aqueous extracts, ethanol extracts, or oils of the targeted plant species and colloidal silver were prepared or purchased. Activity against the wall-less bacterial pathogen Mycoplasma mycoides subsp. capri was determined and compared to activities measured against Escherichia coli and Bacillus subtilis. Antimicrobial susceptibility testing was performed by broth microdilution assays. The lethal or inhibitory nature of each extract was determined by subculture into neat growth medium. Results Growth of M. mycoides capri, E. coli, and B. subtilis was inhibited by elderberry extract, oregano oil, ethanol extract of oregano leaves, and ethanol extract of goldenseal root. No inhibition was seen with aqueous extract of astragalus or calendula oil. Growth of M. mycoides capri and B. subtilis was inhibited by ethanol extract of astragalus, whereas growth of E. coli was not. Similarly, M. mycoides capri and E. coli were inhibited by aqueous extract of thyme, but B. subtilis was unaffected. Only B. subtilis was inhibited by colloidal silver. Measured MICs ranged from 0.0003 mg/mL to 3.8 mg/mL. Bacteriostatic and bactericidal effects differed by species and extract. Conclusions The atypical pathogen M. mycoides capri was sensitive to extracts from many medicinal plants commonly used as antimicrobials in states of preparation and concentrations currently available for purchase in the United States and Europe. Variation in bacteriostatic and bactericidal activities between species and extracts indicates that multiple effecter compounds are present in these plant species. PMID:23031072

  16. Mycoplasma salivarium as a Dominant Coloniser of Fanconi Anaemia Associated Oral Carcinoma

    PubMed Central

    Henrich, Birgit; Rumming, Madis; Sczyrba, Alexander; Velleuer, Eunike; Dietrich, Ralf; Gerlach, Wolfgang; Gombert, Michael; Rahn, Sebastian; Stoye, Jens; Borkhardt, Arndt; Fischer, Ute

    2014-01-01

    Mycoplasma salivarium belongs to the class of the smallest self-replicating Tenericutes and is predominantly found in the oral cavity of humans. In general it is considered as a non-pathogenic commensal. However, some reports point to an association with human diseases. M. salivarium was found e.g. as causative agent of a submasseteric abscess, in necrotic dental pulp, in brain abscess and clogged biliary stent. Here we describe the detection of M. salivarium on the surface of a squamous cell carcinoma of the tongue of a patient with Fanconi anaemia (FA). FA is an inherited bone marrow failure syndrome based on defective DNA-repair that increases the risk of carcinomas especially oral squamous cell carcinoma. Employing high coverage, massive parallel Roche/454-next-generation-sequencing of 16S rRNA gene amplicons we analysed the oral microbiome of this FA patient in comparison to that of an FA patient with a benign leukoplakia and five healthy individuals. The microbiota of the FA patient with leukoplakia correlated well with that of the healthy controls. A dominance of Streptococcus, Veillonella and Neisseria species was typically observed. In contrast, the microbiome of the cancer bearing FA patient was dominated by Pseudomonas aeruginosa at the healthy sites, which changed to a predominance of 98% M. salivarium on the tumour surface. Quantification of the mycoplasma load in five healthy, two tumour- and two leukoplakia-FA patients by TaqMan-PCR confirmed the prevalence of M. salivarium at the tumour sites. These new findings suggest that this mycoplasma species with its reduced coding capacity found ideal breeding grounds at the tumour sites. Interestingly, the oral cavity of all FA patients and especially samples at the tumour sites were in addition positive for Candida albicans. It remains to be elucidated in further studies whether M. salivarium can be used as a predictive biomarker for tumour development in these patients. PMID:24642836

  17. Sequence TTKF?QE Defines the Site of Proteolytic Cleavage in Mhp683 Protein, a Novel Glycosaminoglycan and Cilium Adhesin of Mycoplasma hyopneumoniae*

    PubMed Central

    Bogema, Daniel R.; Scott, Nichollas E.; Padula, Matthew P.; Tacchi, Jessica L.; Raymond, Benjamin B. A.; Jenkins, Cheryl; Cordwell, Stuart J.; Minion, F. Chris; Walker, Mark J.; Djordjevic, Steven P.

    2011-01-01

    Mycoplasma hyopneumoniae colonizes the ciliated respiratory epithelium of swine, disrupting mucociliary function and inducing chronic inflammation. P97 and P102 family members are major surface proteins of M. hyopneumoniae and play key roles in colonizing cilia via interactions with glycosaminoglycans and mucin. The p102 paralog, mhp683, and homologs in strains from different geographic origins encode a 135-kDa pre-protein (P135) that is cleaved into three fragments identified here as P45683, P48683, and P50683. A peptide sequence (TTKF?QE) was identified surrounding both cleavage sites in Mhp683. N-terminal sequences of P48683 and P50683, determined by Edman degradation and mass spectrometry, confirmed cleavage after the phenylalanine residue. A similar proteolytic cleavage site was identified by mass spectrometry in another paralog of the P97/P102 family. Trypsin digestion and surface biotinylation studies showed that P45683, P48683, and P50683 reside on the M. hyopneumoniae cell surface. Binding assays of recombinant proteins F1683–F5683, spanning Mhp683, showed saturable and dose-dependent binding to biotinylated heparin that was inhibited by unlabeled heparin, fucoidan, and mucin. F1683–F5683 also bound porcine epithelial cilia, and antisera to F2683 and F5683 significantly inhibited cilium binding by M. hyopneumoniae cells. These data suggest that P45683, P48683, and P50683 each display cilium- and proteoglycan-binding sites. Mhp683 is the first characterized glycosaminoglycan-binding member of the P102 family. PMID:21969369

  18. The pyruvate dehydrogenase complex of Mycoplasma hyopneumoniae contains a novel lipoyl domain arrangement

    Microsoft Academic Search

    Jake N. Matic; Jody L. Wilton; Rebecca J. Towers; Anthony L. Scarman; F. Chris Minion; Mark J. Walker; Steve P. Djordjevic

    2003-01-01

    The genes encoding the pyruvate dehydrogenase (PDH) complex (pdhA, pdhB, pdhC and pdhD) from Mycoplasma hyopneumoniae have been cloned and sequenced. The genes are arranged into two operons, designated pdhAB and pdhCD, which are not found together in the chromosome. The pdhA, pdhB, pdhC and pdhD genes encode proteins of predicted molecular masses of 44.2 kDa (pyruvate dehydrogenase major subunit;

  19. Mycoplasma gallisepticum infection in the grey partridge Perdix perdix : outbreak description, histopathology, biochemistry and antioxidant parameters

    Microsoft Academic Search

    Frantisek Vitula; Lucie Peckova; Hana Bandouchova; Miroslav Pohanka; Ladislav Novotny; David Jira; Jiri Kral; Karel Ondracek; Jitka Osickova; Dagmar Zendulkova; Katerina Rosenbergova; Frantisek Treml; Jiri Pikula

    2011-01-01

    Background  The grey partridge is an important game bird in Europe that has declined considerably over the last decades. The production\\u000a and release of farm-bred birds can be threatened by infectious agents. The objective of this study was to describe the outbreak,\\u000a pathology, and blood and tissue biochemical responses in a flock of grey partridges naturally infected with Mycoplasma gallisepticum.\\u000a \\u000a \\u000a \\u000a \\u000a Results  Morbidity

  20. Coexistence of Pulmonary Hydatid Cyst and Mycoplasma pneumoniae Pnömonia in a Child.

    PubMed

    Gayretli Ayd?n, Zeynep Gökçe; Yalç?nkaya, Rümeysa; Ayd?n Teke, Türkan; Bayhan, Gülsüm ?clal; Öz, Fatma Nur; Metin Timur, Özge; Ek?io?lu, Ay?e Seçil; Tan?r, Gönül

    2015-06-01

    Hydatid cyst is a zoonotic disease and endemic in Turkey. The disease can involve any organ. The most common involved organ is lung in childhood. Hydatid cyst of lung may be asymptomatic or may be sometimes ruptured or infected. Secondary bacterial infections associated with the hydatid cyst are well known. A previously not reported pediatric case of hydatid cyst with Mycoplasma pneumoniae pneumonia is described in this report. It is emphasized that M. pneumoniae should be kept in mind as a cause of infected hydatid cyst which is unresponsive to beta-laktam antibiotics. PMID:26081892

  1. Histopathological and immunohistochemical findings in the lungs of pigs infected experimentally with Mycoplasma hyopneumoniae.

    PubMed

    Redondo, E; Masot, A J; Fernández, A; Gázquez, A

    2009-05-01

    Pigs were infected intranasally with Mycoplasma hyopneumoniae and killed at intervals ranging from 7 to 35 days post-infection (dpi). Histopathological changes consisted of (1) exudates in airways and alveolar lumina, (2) peribronchial and peribronchiolar lymphoid hyperplasia, and (3) enlargement of alveolar septa. These changes were particularly marked from 7 to 28dpi, coinciding with significant increases in the expression, detected immunohistochemically, of cytokines (IL-1alpha, IL-1beta, IL-8, TNF-alpha and INF-gamma) and lymphoid markers (CD4+, CD8+, muramidase, IgG+, IgA+). Both the lesions and immunohistochemical signals declined in intensity beyond 35 days. PMID:19285314

  2. Kinetics of Sodium Dodecyl Sulfate Solubilization of Mycoplasma laidlawii Plasma Membranes

    PubMed Central

    Auborn, James J.; Eyring, Edward M.; Choules, G. Lew

    1971-01-01

    The kinetics of sodium dodecyl sulfate solubilization of aqueous suspensions of Mycoplasma laidlawii membranes have been investigated by light scattering in a stopped-flow apparatus. There was evidence of direct interaction between the membranes and sodium dodecyl sulfate micelles above the critical micelle concentration, although of lower order kinetically than with monomeric dodecyl sulfate anions below the critical micelle concentration. The activation energy remained the same in either case, about 10 kcal/mol. Static light-scattering studies at higher resolution showed that the solubilized membranes are in the form of small aggregates. PMID:5289357

  3. Chlamydia pneumoniae and mycoplasma pneumoniae in children with acute respiratory infection in general practices in the Netherlands

    Microsoft Academic Search

    J. H. T. Tjhie; J. W. Dorigo-Zetsma; R. Roosendaal; T. M. Bestebroer; A. I. M. Bartelds; C. M. J. E. Vandenbroucke-Grauls

    2000-01-01

    In this retrospective study Chlamydia pneumoniae and Mycoplasma pneumoniae infections were detected by polymerase chain reaction (PCR) in samples (n=457) from children presenting with acute respiratory infection to general practitioners during 1992-97. Samples were collected in autumn and winter, and from 1994 onwards in spring and summer also. Overall, C. pneumoniae and M. pneumoniae were detected in throat or nasal

  4. Antibiotic prescriptions and cycles of Mycoplasma pneumoniae infections in Norway: can a nationwide prescription register be used for surveillance?

    PubMed

    Blix, H S; Vestrheim, D F; Hjellvik, V; Skaare, D; Christensen, A; Steinbakk, M

    2015-07-01

    Mycoplasma pneumoniae outbreaks cause increased use of macrolides and tetracyclines. We aimed to investigate whether drug use data, in addition to laboratory data, could improve understanding of the spread of M. pneumoniae epidemics. Number of users of Mycoplasma antibiotics (erythromycin, doxycycline, clarithromycin) per week and county of residence in an indicator age group (6-12 years) was retrieved from the Norwegian prescription database for the epidemic season 2011-2012 and compared to non-epidemic seasons. In 2011, increased use of Mycoplasma antibiotics was first observed in September on the west coast of Norway. The Norwegian laboratory-based surveillance system showed the first increase in positive tests in August 2011 and an epidemic was announced on 25 October 2011. At that time the use of Mycoplasma antibiotics had already exceeded three times the use in non-epidemic periods. Data for three counties from the regional microbiological laboratories showed that the increase in number of positive samples coincided in time with the increase in prescription data. Laboratory data cannot accurately determine the extent of an epidemic, and drug use data cannot identify the cause. Establishing a systematic interaction between the two monitoring systems will enhance surveillance and probably contribute to improved infection control and prudent antibiotic prescribing. PMID:25388750

  5. Evaluation of enzyme-linked immunosorbent assays for detection of Mycoplasma Bovis-specific antibody in bison sera

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bovis has recently emerged as a significant and costly infectious disease problem in bison. This report demonstrates that ELISAs for detection of M. bovis-specific antibody in cattle are not optimal for identification of seropositive bison. An ELISA optimized for use with bison sera is ...

  6. Erythema Nodosum and Mycoplasma pneumoniae Infections in Childhood: Further Observations in Two Patients and a Literature Review

    PubMed Central

    Greco, Filippo; Catania, Roberta; Pira, Alice Le; Saporito, Marco; Scalora, Luisa; Aguglia, Maria Giovanna; Smilari, Pierluigi; Sorge, Giovanni

    2015-01-01

    Erythema nodosum (EN) is the most frequent panniculitis in childhood and has been associated with various conditions, such as infectious and autoimmune disorders, medications, and malignancies. The author reports on two children affected with EN associated with Mycoplasma pneumoniae infection, which occurred in one patient without pulmonary detection. The available literature on EN and M. pneumoniae infection in childhood is also reviewed. PMID:25699127

  7. 9 CFR 147.16 - Procedure for the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    Research results are described in the following two publications: (a) Bigland, C. H. and A. J. DaMassa, “A Bio-Assay for Mycoplasma Gallisepticum.” In: United States Livestock Sanitary Association Proceedings, 67th, 1963, pp. 541-549. (b) McMartin, D....

  8. Survival of bighorn sheep (Ovis canadensis) commingled with domestic sheep (Ovis aries) in the absence of mycoplasma ovipneumoniae.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To test the hypothesis that Mycoplasma ovipneumoniae is an important agent of the bighorn sheep (Ovis canadensis) pneumonia that has previously inevitably followed experimental commingling with domestic sheep (Ovis aries), we commingled M. ovipneumoniae–free domestic and bighorn sheep (n=4 each). On...

  9. Direct and indirect calorimetric studies of stress responses of chlorella cells to infection with the mycoplasma, Acholeplasma laidlawii

    Microsoft Academic Search

    N. L. Loseva; L. K. Gordon; F. V. Minibayeva; A. Ju. Alyabyev; V. M. Chernov; O. A. Chernova; I. N. Andreyeva; G. G. Rachimova; V. I. Tribunskih; R. I. Estrina; Ju. B. Gogolev; R. B. Kemp

    2002-01-01

    This paper reports the defence responses of plant cells to the stress of infection by mycoplasma using an algae model of Chlorella vulgaris under attack by the Mollicute, Acholeplasma laidlawii, which is normally a pathogen of animal systems and higher plants. When the two unicellular organisms were mixed, there was a significant rise in the heat flow rate from 30min

  10. Cloning and functional analysis of the P97 swine cilium adhesin gene of Mycoplasma hyopneumoniae.

    PubMed Central

    Hsu, T; Artiushin, S; Minion, F C

    1997-01-01

    Colonization of the swine respiratory tract by Mycoplasma hyopneumoniae is accomplished by specific binding to the cilia of the mucosal epithelial cells. Previous studies have implicated a 97-kDa outer membrane-associated protein, P97, that appeared to mediate this interaction. In order to further define the role of P97 in adherence to porcine cilia, the structural gene was cloned and sequenced, and the recombinant products were analyzed. Monoclonal antibodies were used to identify recombinant clones in a genomic library expressed in an opal suppressor host because of alternate codon usage by mycoplasmas. The gene coding for P97 was then identified by Tn1000 mutagenesis of recombinant clones. DNA sequence analysis revealed an open reading frame coding for a 124.9-kDa protein with a hydrophobic transmembrane spanning domain. The N-terminal sequence of purified P97 mapped at amino acid position 195 of the translated sequence, indicating that a processing event had occurred in M. hyopneumoniae. Both recombinant P97 protein expressed in an Escherichia coli opal suppressor host and M. hyopneumoniae bound specifically to swine cilia, and the binding was inhibited by heparin and fucoidan, thus supporting the hypothesis that P97 was actively involved in binding to swine cilia in vivo. PMID:9023217

  11. Neutrophil Dependence of Vascular Remodeling after Mycoplasma Infection of Mouse Airways

    PubMed Central

    Baluk, Peter; Phillips, Keeley; Yao, Li-Chin; Adams, Alicia; Nitschké, Maximilian; McDonald, Donald M.

    2015-01-01

    Vascular remodeling is a feature of sustained inflammation in which capillaries enlarge and acquire the phenotype of venules specialized for plasma leakage and leukocyte recruitment. We sought to determine whether neutrophils are required for vascular remodeling in the respiratory tract by using Mycoplasma pulmonis infection as a model of sustained inflammation in mice. The time course of vascular remodeling coincided with the influx of neutrophils during the first few days after infection and peaked at day 5. Depletion of neutrophils with antibody RB6-8C5 or 1A8 reduced neutrophil influx and vascular remodeling after infection by about 90%. Similarly, vascular remodeling after infection was suppressed in Cxcr2?/? mice, in which neutrophils adhered to the endothelium of venules but did not extravasate into the tissue. Expression of the venular adhesion molecule P-selectin increased in endothelial cells from day 1 to day 3 after infection, as did expression of the Cxcr2-receptor ligands Cxcl1 and Cxcl2. Tumor necrosis factor ? (TNF?) expression increased more than sixfold in the trachea of wild-type and Cxcr2?/? mice, but intratracheal administration of TNF? did not induce vascular remodeling similar to that seen in infection. We conclude that neutrophil influx is required for remodeling of capillaries into venules in the airways of mice with Mycoplasma infection and that TNF? signaling is necessary but not sufficient for vascular remodeling. PMID:24726646

  12. Late lesions of experimental contagious caprine pleuropneumonia caused by Mycoplasma capricolum ssp. capripneumoniae.

    PubMed

    Wesonga, H O; Lindberg, R; Litamoi, J K; Bölske, G

    1998-03-01

    A clinical, bacteriological, serological and patho-anatomical study was carried out on 12 goats surviving the acute stage of contagious caprine pleuropneumonia (CCPP), experimentally produced with Mycoplasma capricolum ssp. capripneumoniae (M. capripneumoniae), with the major aims of investigating the chronic stage of the disease and elucidating the possibility of a carrier state beyond the acute fulminant phase. The goats were killed 9, 16, 82 or 126 days after the onset of acute clinical signs. On day 9, clinical signs included low grade fever and persistent coughing. Thereafter, only intermittent coughing was recorded. Serum titres of complement-fixing antibodies to M. capripneumoniae were high at the period of fever but dropped thereafter. Post-mortem examination showed acute fibrinous pleuropneumonia on days 9 and 16, and chronic pleuropneumonia on days 82 and 126, including sequester formations in goats killed on day 126. Mycoplasma capripneumoniae was isolated on days 9 and 16 but not on later occasions. The study showed that goats recovered from acute CCPP may have lesions for a long time thereafter but provide no evidence of a carrier state among long-term survivors. PMID:9557132

  13. Invasion and persistence of Mycoplasma bovis in embryonic calf turbinate cells.

    PubMed

    Bürki, Sibylle; Gaschen, Véronique; Stoffel, Michael H; Stojiljkovic, Ana; Frey, Joachim; Kuehni-Boghenbor, Kathrin; Pilo, Paola

    2015-01-01

    Mycoplasma bovis is a wall-less bacterium causing bovine mycoplasmosis, a disease showing a broad range of clinical manifestations in cattle. It leads to enormous economic losses to the beef and dairy industries. Antibiotic treatments are not efficacious and currently no efficient vaccine is available. Moreover, mechanisms of pathogenicity of this bacterium are not clear, as few virulence attributes are known. Microscopic observations of necropsy material suggest the possibility of an intracellular stage of M. bovis. We used a combination of a gentamicin protection assay, a variety of chemical treatments to block mycoplasmas entry in eukaryotic cells, and fluorescence and transmission electron microscopy to investigate the intracellular life of M. bovis in calf turbinate cells. Our findings indicate that M. bovis invades and persists in primary embryonic calf turbinate cells. Moreover, M. bovis can multiply within these cells. The intracellular phase of M. bovis may represent a protective niche for this pathogen and contribute to its escape from the host's immune defense as well as avoidance of antimicrobial agents. PMID:25976415

  14. Mycoplasma genitalium Infection and Female Reproductive Tract Disease: A Meta-analysis.

    PubMed

    Lis, Rebecca; Rowhani-Rahbar, Ali; Manhart, Lisa E

    2015-08-01

    To determine the association between Mycoplasma genitalium infection and female reproductive tract syndromes through meta-analysis, English-language, peer-reviewed studies were identified via PubMed, Embase, Biosis, Cochrane Library, and reference review. Two reviewers independently extracted data. Random-effects models were employed to calculate summary estimates, between-study heterogeneity was evaluated using I(2) statistics, publication bias was assessed via funnel plots and the Begg and Egger tests, and methodologic quality was rated. Mycoplasma genitalium infection was significantly associated with increased risk of cervicitis (pooled odds ratio [OR], 1.66 [95% confidence interval {CI}, 1.35-2.04]), pelvic inflammatory disease (pooled OR, 2.14 [95% CI, 1.31-3.49]), preterm birth (pooled OR, 1.89 [95% CI, 1.25-2.85]), and spontaneous abortion (pooled OR, 1.82 [95% CI, 1.10-3.03]). Risk of infertility was similarly elevated (pooled OR, 2.43 [95% CI, .93-6.34]). In subanalyses accounting for coinfections, all associations were stronger and statistically significant. Testing of high-risk symptomatic women for M. genitalium may be warranted. PMID:25900174

  15. 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.

  16. A novel sigma factor reveals a unique regulon controlling cell-specific recombination in Mycoplasma genitalium.

    PubMed

    Torres-Puig, Sergi; Broto, Alicia; Querol, Enrique; Piñol, Jaume; Pich, Oscar Q

    2015-05-26

    The Mycoplasma genitalium MG428 protein shows homology to members of the sigma-70 family of sigma factors. Herein, we found that MG428 activates transcription of recA, ruvA and ruvB as well as several genes with unknown function. Deletion of MG_428 or some of the up-regulated unknown genes led to severe recombination defects. Single cell analyses revealed that activation of the MG428-regulon is a rare event under laboratory growth conditions. A conserved sequence with sigma-70 promoter architecture (TTGTCA-N18/19-ATTWAT) was identified in the upstream region of all of the MG428-regulated genes or operons. Primer extension analyses demonstrated that transcription initiates immediately downstream of this sigma70-type promoter in a MG428-dependent manner. Furthermore, mutagenesis of the conserved -10 and -35 elements corroborated the requirement of these regions for promoter function. Therefore, a new mycoplasma promoter directs transcription of a unique recombination regulon. Additionally, MG428 was found to interact with the RNAP core enzyme, reinforcing the predicted role of this protein as an alternative sigma factor. Finally, our results indicate that MG428 contributes to the generation of genetic diversity in this model organism. Since recombination is an important mechanism to generate antigenic variation, MG428 emerges as a novel factor contributing to M. genitalium virulence. PMID:25925568

  17. Number of specific antibody-secreting cells in the peripheral blood among children with mycoplasma pneumonia.

    PubMed Central

    Iseki, M; Takahashi, T; Kimura, K; Yamashita, R; Sasaki, T

    1996-01-01

    Mycoplasma pneumoniae-specific antibody-secreting cells (ASCs) in the peripheral blood were enumerated with an enzyme-linked immunospot assay in 12 children with mycoplasma pneumonia. Those cells were detected in the acute phases and declined in number in the convalescent stage. The maximum numbers of M. pneumoniae-specific ASCs ranged from 0 to 478 for immunoglobulin G (IgG), 13 to 1,992 for IgM, and 0 to 53 for IgA per 106 peripheral blood mononuclear cells, whereas the total numbers (i.e., including both specific and nonspecific) of immunoglobulin-secreting cells (IgSCs) were as high as 4,000 for both IgG and IgM and 1,000 for IgA per 106 peripheral blood mononuclear cells. Such a great increase in the numbers of total IgSCs in comparison with that in M. pneumoniae-specific ASCs suggests that the majority of the IgSC increase in the course of mycoplasmal infection was nonspecific to M. pneumoniae. The serum level of M. pneumoniae antibody measured by enzyme-linked immunosorbent assay remained high in the convalescent phase, while the number of specific ASCs decreased. Whereas this observation may be explained by declined degeneration or consumption of the antibody in the convalescent phase, it may be suggestive of the source of M. pneumoniae antibody other than ASCs in the peripheral blood. PMID:8698511

  18. Domain Analysis of Protein P30 in Mycoplasma pneumoniae Cytadherence and Gliding Motility? ‡

    PubMed Central

    Chang, How-Yi; Jordan, Jarrat L.; Krause, Duncan C.

    2011-01-01

    The cell wall-less prokaryote Mycoplasma pneumoniae causes bronchitis and atypical pneumonia in humans. Mycoplasma attachment and gliding motility are required for colonization of the respiratory epithelium and are mediated largely by a differentiated terminal organelle. P30 is a membrane protein at the distal end of the terminal organelle and is required for cytadherence and gliding motility, but little is known about the functional role of its specific domains. In the current study, domain deletion and substitution derivatives of P30 were engineered and introduced into a P30 null mutant by transposon delivery to assess their ability to rescue P30 function. Domain deletions involving the extracellular region of P30 severely impacted protein stability and adherence and gliding function, as well as the capacity to stabilize terminal organelle protein P65. Amino acid substitutions in the transmembrane domain revealed specific residues uniquely required for P30 stability and function, perhaps to establish correct topography in the membrane for effective alignment with binding partners. Deletions within the predicted cytoplasmic domain did not affect P30 localization or its capacity to stabilize P65 but markedly impaired gliding motility and cytadherence. The larger of two cytoplasmic domain deletions also appeared to remove the P30 signal peptide processing site, suggesting a larger leader peptide than expected. We propose that the P30 cytoplasmic domain may be required to link P30 to the terminal organelle core, to enable the P30 extracellular domain to achieve a functional conformation, or perhaps both. PMID:21257768

  19. Domain analysis of protein P30 in Mycoplasma pneumoniae cytadherence and gliding motility.

    PubMed

    Chang, How-Yi; Jordan, Jarrat L; Krause, Duncan C

    2011-04-01

    The cell wall-less prokaryote Mycoplasma pneumoniae causes bronchitis and atypical pneumonia in humans. Mycoplasma attachment and gliding motility are required for colonization of the respiratory epithelium and are mediated largely by a differentiated terminal organelle. P30 is a membrane protein at the distal end of the terminal organelle and is required for cytadherence and gliding motility, but little is known about the functional role of its specific domains. In the current study, domain deletion and substitution derivatives of P30 were engineered and introduced into a P30 null mutant by transposon delivery to assess their ability to rescue P30 function. Domain deletions involving the extracellular region of P30 severely impacted protein stability and adherence and gliding function, as well as the capacity to stabilize terminal organelle protein P65. Amino acid substitutions in the transmembrane domain revealed specific residues uniquely required for P30 stability and function, perhaps to establish correct topography in the membrane for effective alignment with binding partners. Deletions within the predicted cytoplasmic domain did not affect P30 localization or its capacity to stabilize P65 but markedly impaired gliding motility and cytadherence. The larger of two cytoplasmic domain deletions also appeared to remove the P30 signal peptide processing site, suggesting a larger leader peptide than expected. We propose that the P30 cytoplasmic domain may be required to link P30 to the terminal organelle core, to enable the P30 extracellular domain to achieve a functional conformation, or perhaps both. PMID:21257768

  20. Processing Is Required for a Fully Functional Protein P30 in Mycoplasma pneumoniae Gliding and Cytadherence ?

    PubMed Central

    Chang, How-Yi; Prince, Oliver A.; Sheppard, Edward S.; Krause, Duncan C.

    2011-01-01

    The cell wall-less prokaryote Mycoplasma pneumoniae causes bronchitis and atypical pneumonia in humans. Mycoplasma attachment to the host respiratory epithelium is required for colonization and mediated largely by a differentiated terminal organelle. P30 is an integral membrane protein located at the distal end of the terminal organelle. The P30 null mutant II-3 is unable to attach to host cells and nonmotile and has a branched cellular morphology compared to the wild type, indicating an important role for P30 in M. pneumoniae biology. P30 is predicted to have an N-terminal signal sequence, but the presence of such a motif has not been confirmed experimentally. In the current study we analyzed P30 derivatives having epitope tags engineered at various locations to demonstrate that posttranslational processing occurred in P30. Several potential cleavage sites predicted in silico were examined, and a processing-defective mutant was created to explore P30 maturation further. Our results suggested that signal peptide cleavage occurs between residues 52 and 53 to yield mature P30. The processing-defective mutant exhibited reduced gliding velocity and cytadherence, indicating that processing is required for fully functional maturation of P30. We speculate that P30 processing may trigger a conformational change in the extracellular domain or expose a binding site on the cytoplasmic domain to allow interaction with a binding partner as a part of functional maturation. PMID:21821772

  1. Processing is required for a fully functional protein P30 in Mycoplasma pneumoniae gliding and cytadherence.

    PubMed

    Chang, How-Yi; Prince, Oliver A; Sheppard, Edward S; Krause, Duncan C

    2011-10-01

    The cell wall-less prokaryote Mycoplasma pneumoniae causes bronchitis and atypical pneumonia in humans. Mycoplasma attachment to the host respiratory epithelium is required for colonization and mediated largely by a differentiated terminal organelle. P30 is an integral membrane protein located at the distal end of the terminal organelle. The P30 null mutant II-3 is unable to attach to host cells and nonmotile and has a branched cellular morphology compared to the wild type, indicating an important role for P30 in M. pneumoniae biology. P30 is predicted to have an N-terminal signal sequence, but the presence of such a motif has not been confirmed experimentally. In the current study we analyzed P30 derivatives having epitope tags engineered at various locations to demonstrate that posttranslational processing occurred in P30. Several potential cleavage sites predicted in silico were examined, and a processing-defective mutant was created to explore P30 maturation further. Our results suggested that signal peptide cleavage occurs between residues 52 and 53 to yield mature P30. The processing-defective mutant exhibited reduced gliding velocity and cytadherence, indicating that processing is required for fully functional maturation of P30. We speculate that P30 processing may trigger a conformational change in the extracellular domain or expose a binding site on the cytoplasmic domain to allow interaction with a binding partner as a part of functional maturation. PMID:21821772

  2. Isolation and Characterization of Fractions of Mycoplasma pneumoniae I. Chemical and Chromatographic Separation

    PubMed Central

    Prescott, B.; Sobeslavsky, O.; Caldes, G.; Chanock, R. M.

    1966-01-01

    Prescott, B. (National Institute of Allergy and Infectious Diseases, Bethesda, Md.), O. Sobeslavsky, G. Caldes, and R. M. Chanock. Isolation and characterization of fractions of Mycoplasma pneumoniae. I. Chemical and chromatographic separation. J. Bacteriol. 91:2117–2125. 1966.—Fractionation of Mycoplasma pneumoniae, cultured on a beef heart infusion-horse serum-yeast extract medium, was carried out by chemical and chromatographic procedures. The chemical method yielded eight fractions consisting of lipid, carbohydrates, and proteins. Four protein-rich fractions were isolated by chromatographing a supernatant fluid of sonically treated organisms on Sephadex G-25. The 12 fractions were tested for serological and antigenic activity in vitro and in vivo. The lipid fraction was serologically active and the relative order of activity of the protein fractions appeared to depend on the amount of lipid present in the molecule. The highly serologically active Sephadex G-25 protein fraction 1 prepared chromatographically contained 15% lipid in the molecule, whereas the less serologically active protein fraction 2 prepared by chemical means contained 2% lipid. The acetone-extracted lipid fraction was chromatographed on thin-layer chromatography plates and found to consist of nine fractions. Serological activity was associated with only the first three spots above the origin. Lipid extracted from the protein fractions seemed to be similar to the acetone-extracted lipid from the sediment of the sonically treated organisms. PMID:5943931

  3. Natural Resolution of Air Sac Lesions Caused by Mycoplasma meleagridis in Turkeys

    PubMed Central

    Bigland, C. H.

    1969-01-01

    Lesions caused by Mycoplasma meleagridis were followed in one hatch of turkeys from one day to 20½ weeks of age. Air sac lesion incidence increased from 20% at one day to 44% at six weeks of age with a slow decline to 10% at 10 weeks of age. Lesions were detectable on careful examination at 10-20% level to 20 weeks of age on laboratory examination, but at 20½ weeks only 0.07% lesions were found in 1000 turkeys from the same hatch. Air sac lesion severity, based on an arbitrary 0.5,1,2,3,+scale, increased in intensity and spread of air sac involvement from a mean of 1.5 at day old to 2.5 at five weeks of age. This declined to 1.0 at 10 weeks of age and lesions became progressively more faint until careful examination was necessary to detect them in birds 15 weeks and older. The latter would not be considered as a cause of total condemnation. The percentage of Mycoplasma isolations from the respiratory tissues were much higher in birds exhibiting air sac lesions than those in which no typical air sac lesions were visible, as follows. air sacs 51.6 vs. 2.9, lungs 43.3 vs. 4.0, trachea 47.2 vs. 4.6, sinus 38.5 vs. 19.0. PMID:4242766

  4. In Vivo Pharmacokinetic/Pharmacodynamic Profiles of Valnemulin in an Experimental Intratracheal Mycoplasma gallisepticum Infection Model.

    PubMed

    Xiao, Xia; Sun, Jian; Yang, Tao; Fang, Xi; Wu, Dong; Xiong, Yan Q; Cheng, Jie; Chen, Yi; Shi, Wei; Liu, Ya-Hong

    2015-07-01

    Valnemulin, a semisynthetic pleuromutilin antibiotic derivative, is greatly active against Mycoplasma. The objective of our study was to evaluate the effectiveness of valnemulin against Mycoplasma gallisepticum in a neutropenic intratracheal model in chickens using a pharmacokinetic/pharmacodynamic (PK-PD) method. The PK of valnemulin after intramuscular (i.m.) administration at doses of 1, 10, and 20 mg/kg of body weight in M. gallisepticum-infected neutropenic chickens was evaluated by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Real-time PCR (RT-PCR) was used for quantitative detection of M. gallisepticum. The ratio of the 24-h area under the concentration-time curve divided by the MIC (AUC24/MIC) correlated well with the in vivo antibacterial effectiveness of valnemulin (R(2) = 0.9669). The AUC24/MIC ratios for mycoplasmastasis (a reduction of 0 log10 color-changing unit [CCU] equivalents/ml), a reduction of 1 log10 CCU equivalents/ml, and a reduction of 2.5 log10 CCU equivalents/ml are 28,820, 38,030, and 56,256, respectively. In addition, we demonstrated that valnemulin at a dose of 6.5 mg/kg resulted in a reduction of 2.5 log10 CCU equivalents/ml. These investigations provide a solid foundation for the usage of valnemulin in poultry with M. gallisepticum infections. PMID:25845865

  5. A novel sigma factor reveals a unique regulon controlling cell-specific recombination in Mycoplasma genitalium

    PubMed Central

    Torres-Puig, Sergi; Broto, Alicia; Querol, Enrique; Piñol, Jaume; Pich, Oscar Q.

    2015-01-01

    The Mycoplasma genitalium MG428 protein shows homology to members of the sigma-70 family of sigma factors. Herein, we found that MG428 activates transcription of recA, ruvA and ruvB as well as several genes with unknown function. Deletion of MG_428 or some of the up-regulated unknown genes led to severe recombination defects. Single cell analyses revealed that activation of the MG428-regulon is a rare event under laboratory growth conditions. A conserved sequence with sigma-70 promoter architecture (TTGTCA-N18/19-ATTWAT) was identified in the upstream region of all of the MG428-regulated genes or operons. Primer extension analyses demonstrated that transcription initiates immediately downstream of this sigma70-type promoter in a MG428-dependent manner. Furthermore, mutagenesis of the conserved ?10 and ?35 elements corroborated the requirement of these regions for promoter function. Therefore, a new mycoplasma promoter directs transcription of a unique recombination regulon. Additionally, MG428 was found to interact with the RNAP core enzyme, reinforcing the predicted role of this protein as an alternative sigma factor. Finally, our results indicate that MG428 contributes to the generation of genetic diversity in this model organism. Since recombination is an important mechanism to generate antigenic variation, MG428 emerges as a novel factor contributing to M. genitalium virulence. PMID:25925568

  6. 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

  7. Mouse Repository Strain Details

    Cancer.gov

    Available Strain Details Order Form for Cryoarchived Strains   Strain Number: 01XB8  Common Strain Name: Brca1 floxed (FVB;129)  Strain Nomenclature: FVB;129-Brca1tm1Brn/Nci  Release Category (Required for MTA form): C2 , D Sample MTA for this strain Strain

  8. Linezolid-resistant Staphylococcus haemolyticus and Staphylococcus hominis: single and double mutations at the domain V of 23S rRNA among isolates from a Rio de Janeiro hospital.

    PubMed

    Chamon, Raiane Cardoso; Iorio, Natalia Lopes Pontes; Cavalcante, Fernanda Sampaio; Teodoro, Cristiane R S; de Oliveira, Ana Paula Chaves; Maia, Fernanda; dos Santos, Kátia Regina Netto

    2014-12-01

    In this work, the molecular and phenotypic antimicrobial resistance and clonal diversity of 10 linezolid-resistant Staphylococcus spp. isolates were investigated. The 7 Staphylococcus haemolyticus isolates presented Staphylococcal cassete chromosome mec (SCCmec) V and belonged to the same pulsed-field gel electrophoresis pulsotype. Their MICs for oxacillin, vancomycin, and linezolid were ? 256 ?g/mL, 1-4 ?g/mL, and 8-16 ?g/mL, respectively. The 3 S. hominis presented MIC values 32 to >256 ?g/mL, 2-4 ?g/mL, and 12-24 ?g/mL, and all carried the nontypeable SCCmec (ccr1 + mecA class) and belonged to 2 different genotypes. The cfr gene was not found, but the mutation G2603T was detected in S. haemolyticus and C2190T and G2603T in Staphylococcus hominis in 23S rRNA. This study demonstrates the spread of a linezolid-resistant S. haemolyticus genotype and, for the first time, describes the mutation C2190T among S. hominis isolates with a double mutation in Brazil. PMID:25294302

  9. Microsporidia and Cryptosporidium in horses and donkeys in Algeria: detection of a novel Cryptosporidium hominis subtype family (Ik) in a horse.

    PubMed

    Laatamna, Abd Elkarim; Wagnerová, Pavla; Sak, Bohumil; Kv?to?ová, Dana; Xiao, Lihua; Rost, Michael; McEvoy, John; Saadi, Ahmed Rachid; Aissi, Meriem; Kvá?, Martin

    2015-03-15

    A total of 219 and 124 individual fecal samples of horses and donkeys, respectively, were screened for the presence of Cryptosporidium spp., Encephalitozoon spp., and Enterocytozoon bieneusi DNA by genus-specific nested PCR. Isolates were genotyped by sequence analysis of SSU rRNA, GP60, TRAP-C1, COWP, and HSP70 loci in Cryptosporidium, and the ITS region in microsporidia. Cryptosporidium spp. was detected on 3/18 horse farms and 1/15 farms where donkeys were kept. Overall, five (2.3%) horse and two (1.6%) donkey specimens were PCR positive for Cryptosporidium. Genotyping at SSU and GP60 loci revealed that three isolates from horses and donkeys were C. parvum subtype family IIaA16G1R1, one isolate from a horse was, C. muris RN66, and one isolate from a donkey was C. muris TS03. An isolate from a horse shared 99.4% and 99.3% similarity with Cryptosporidium hominis and C. cuniculus, respectively, at the SSU locus. This isolate shared 100% identity with C. hominis at the TRAP-C1, COWP, and HSP70 loci, and it was from the novel gp60 subtype family IkA15G1. Microsporidia were found on 6/18 horse and 2/15 donkey farms. E. bieneusi was identified in 6.8% (15/219) and 1.6% (2/124), and Encephalitozoon cuniculi was identified in 1.8% (4/219) and 1.6% (2/124), of horses and donkeys, respectively. Three genotypes of E. cuniculi (I, II and III) were detected in horses, and E. cuniculi genotype II was detected in donkeys. Four genotypes of E. bieneusi (horse1, horse 2, CZ3, D) were described in horses. An additional five horses and two donkeys were positive for E. bieneusi, but the isolated were not genotyped. Neither Cryptosporidium nor microsporidia prevalence were affected by sex, age, type of breeding, or whether the host was a horse or a donkey. PMID:25638716

  10. Effects of different vaccine combinations against Mycoplasma gallisepticum on the internal egg and eggshell characteristics of commercial layer chickens1,2,3.

    PubMed

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

    2015-05-01

    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, but afford lower levels of protection. This has led to research investigating their use in combination with a subsequent overlay vaccination of FMG given later in the production cycle. In the present study, 2 trials were conducted to investigate the effects of prelay vaccinations of live and killed MG vaccines or their combination, in conjunction with an FMG vaccine overlay after peak production, on the egg characteristics of commercial layers. The following vaccination treatments were administered at 10 wk of age (woa): 1) unvaccinated (Control), 2) MG-Bacterin (MGBac) vaccine, 3) ts-11 strain MG (ts11MG) vaccine, and 4) MGBac and ts11MG combination (MGBac + ts11MG). At 45 woa, half of the birds were overlaid with an FMG vaccine. In each trial, internal egg and eggshell parameters including egg weight (EW), Haugh unit score (HU), eggshell breaking strength (EBS), percentage yolk weight (PYW), percentage albumen weight (PAW), percentage eggshell weight (PSW), eggshell weight per unit surface area (SWUSA), percentage yolk moisture (PYM), and percent total lipids (PYL) were determined at various time periods between 21 and 52 woa. At 28 woa, SWUSA was lower in the ts11MG and MGBac + ts11MG groups compared to the Control group. Conversely, at 43 woa, SWUSA was higher in the ts11MG than in the MGBac group. Between 23 and 43 woa, PYL was higher in the MGBac and ts11MG groups in comparison to the Control group. In conclusion, vaccination with MGBac alone or in combination with ts11MG at 10 woa with or without an FMG vaccine overlay at 45 woa does not adversely affect the internal egg or eggshell quality of commercial layers throughout lay. PMID:25701207

  11. Macroscopic haemoglobinuria associated with Mycoplasma pneumoniae infection successfully treated by clarithromycin.

    PubMed

    Kazama, Itsuro; Tamada, Tsutomu; Nakajima, Toshiyuki

    2015-03-01

    A 25-year-old man developed macroscopic haemoglobinuria after a persistent dry cough. Although chest radiograph findings were normal, since the serum antibody for Mycoplasma pneumoniae was significantly elevated, a diagnosis infection with this organism was made. Despite the absence of apparent anaemia, a marked increase in serum haemolytic markers and positive result for urine haemoglobin indicated the haemolysis of red blood cells, which was likely to have occurred secondarily to M. pneumoniae infection. Shortly after the initiation of a macrolide antibiotic, clarithromycin, the patient's haemoglobinuria completely disappeared together with a complete resolution of his respiratory symptoms. In this case, due to the lymphocyte-stimulatory nature of M. pneumoniae, an enhanced immune response, such as the production of cold agglutinins, was likely to be involved in the pathogenesis of erythrocyte haemolysis. The immunomodulatory property of clarithromycin was thought to repress the increased immunological reaction and thus enable the resolution of the urine abnormality. PMID:25819056

  12. Expression and antigenic characterization of recombinant Mycoplasma agalactiae P48 major surface protein.

    PubMed

    Rosati, S; Robino, P; Fadda, M; Pozzi, S; Mannelli, A; Pittau, M

    2000-02-01

    The gene encoding the P48 major surface lipoprotein of M. agalactiae has been recently characterised. Since its product plays an important role in the immune response of infected animals, in this study we analysed a recombinant P48 expressed in E. coli. Multiple point mutations were introduced by site directed mutagenesis in order to convert four tryptophan TGA codons, which are a typical feature of the mycoplasma genetic code, into the standard TGG. The mutated p48 gene was subcloned into pGex-2T and expressed in fusion with glutathione-S transferase. Following purification steps, P48 was eluted from carrier protein by thrombin digestion and used in Western blot and indirect ELISA using well-characterised sheep sera. Results demonstrate that specific antibodies against P48 are detected 3 weeks after onset of clinical disease and the recombinant P48 is a diagnostically relevant marker of M. agalactiae infection. PMID:10703704

  13. Distinguishing between productive and abortive promoters using a random forest classifier in Mycoplasma pneumoniae.

    PubMed

    Lloréns-Rico, Verónica; Lluch-Senar, Maria; Serrano, Luis

    2015-04-20

    Distinguishing between promoter-like sequences in bacteria that belong to true or abortive promoters, or to those that do not initiate transcription at all, is one of the important challenges in transcriptomics. To address this problem, we have studied the genome-reduced bacterium Mycoplasma pneumoniae, for which the RNAs associated with transcriptional start sites have been recently experimentally identified. We determined the contribution to transcription events of different genomic features: the -10, extended -10 and -35 boxes, the UP element, the bases surrounding the -10 box and the nearest-neighbor free energy of the promoter region. Using a random forest classifier and the aforementioned features transformed into scores, we could distinguish between true, abortive promoters and non-promoters with good -10 box sequences. The methods used in this characterization of promoters can be extended to other bacteria and have important applications for promoter design in bacterial genome engineering. PMID:25779052

  14. A Unique Human Mycoplasma Protein that Generically Blocks Antigen-Antibody Union

    PubMed Central

    Nieusma, Travis; Jones, Teresa; Boreo, Isabel; MacLeod, Amanda S.; Mark, Adam; Niessen, Sherry; Kim, Helen J.; Kong, Leopold; Assad-Garcia, Nacyra; Kwon, Keehwan; Chesi, Marta; Smider, Vaughn V.; Salomon, Daniel R.; Jelinek, Diane F.; Kyle, Robert A.; Pyles, Richard B.; Glass, John I.; Ward, Andrew B.; Wilson, Ian A.; Lerner, Richard A.

    2014-01-01

    We report the discovery and crystal structure of a human mycoplasma protein, Protein M, which binds with high affinity to antibodies, predominantly through attachment to the variable region of the ? and ? light chains. Protein M broadly blocks antibody-antigen union and its mechanism of inhibition is of considerable interest because, as a diversity system, the binding mode of each antibody is different. Protein M thus appears to function by a mechanism that is independent of the sequences of members of the extensive antibody repertoire. By anchoring to conserved regions of the antibody light chains, Protein M is in a position to extend its large C-terminal domain over the antibody combining site and block entrance to macromolecular antigens. PMID:24503852

  15. Effect of Lonicera japonica extract on Mycoplasma gallisepticum in naturally infected broiler flocks.

    PubMed

    Mü?tak, H K; Torun, E; Özen, D; Yücel, G; Akan, M; Diker, K S

    2015-06-01

    In this study, the effect of chlorogenic acid extract from Lonicera japonica Thunb. on Mycoplasma gallisepticum infections and the performance of broiler flocks was investigated. A total of 360 Ross-308 broiler chicks taken from M. gallisepticum seropositive flocks were divided equally into three groups designated as control (nothing administered), antibiotic (Tylosin tartrate given for the first 3 d and d 20-22) and test group (chlorogenic acid extract given twice a day on d 16 and 22). Broiler performance analysis, serological tests (slide agglutination), molecular identification (polymerase chain reaction) and histopathological examination were performed to detect M. gallisepticum. The results show that chlorogenic acid not only increases live body weight but is also an alternative treatment option in M. gallisepticum-infected broiler flocks. PMID:25731588

  16. Explaining an Unusually Fast Parasitic Enzyme: Folate Tail-Binding Residues Dictate Substrate Positioning and Catalysis in Cryptosporidium hominis Thymidylate Synthase

    SciTech Connect

    Martucci,W.; Vargo, M.; Anderson, K.

    2008-01-01

    The essential enzyme TS-DHFR from Cryptosporidium hominis undergoes an unusually rapid rate of catalysis at the conserved TS domain, facilitated by two nonconserved residues, Ala287 and Ser290, in the folate tail-binding region. Mutation of these two residues to their conserved counterparts drastically affects multiple steps of the TS catalytic cycle. We have determined the crystal structures of all three mutants (A287F, S290G, and A287F/S290G) in complex with active site ligands dUMP and CB3717. The structural data show two effects of the mutations: an increased distance between the ligands in the active site and increased flexibility of the folate ligand in the partially open enzyme state that precedes conformational change to the active catalytic state. The latter effect is able to be rescued by the mutants containing the A287F mutation. In addition, the conserved water network of TS is altered in each of the mutants. The structural results point to a role of the folate tail-binding residues in closely positioning ChTS ligands and restricting ligand flexibility in the partially open state to allow for a rapid transition to the active closed state and enhanced rate of catalysis. These results provide an explanation on how folate tail-binding residues at one end of the active site affect long-range interactions throughout the TS active site and validate these residues as targets for species-specific drug design.

  17. Detection of serum proteins in the electrophoretic patterns of total proteins of mycoplasma cells.

    PubMed Central

    Yaguzhinskaya, O. E.

    1976-01-01

    The contamination of mycoplasma cell preparations by serum proteins originating from culture medium was studied. A. laidlawii and M. arthritidis cells were grown in the presence of [14C]-aminoacids, and the cells were washed with 0-9% NaC1 by threefold centrifugation. Total proteins of the washed cells were analysed by SDS gel electrophoresis. Coomassie-stained electrophoretic patterns were compared with autoradiographs of the same gels. The stained electrophoretic pattern of washed A. laidlawii grown without serum was identical with autoradiographs of the same cells grown without or with serum. That of washed A. laidlawii grown with serum differed from the corresponding autoradiography by the presence of extra protein bands I, II, III, and IV with molecular weights of over 160,000, 80,000-87,000, 55,000 and 25,000, respectively. The same extra bands were found in stained electrophoretic patterns of washed: (a) A. laidlawii cells grown without serum and mixed with serum in the stationary phase, (b) M. arthritidis cells, as compared with their autoradiographs, (c) serum precipitate. The bands III and IV may be due to the heavy and light chains of gamma-globulin, the band II might belong to transferrin or to some component of complement. Acidification of serum to pH 5 brought about 100-fold rise of amount of serum precipitate, the number of bands in the electrophoretic pattern of the precipitate being also increased. Stained electrophoretic patterns of cells purified by twofold centrifugation in step sucrose density gradient (1-20-1-27 g./cm.3 for A. laidlawii, and 1-15-1-25 for M. arthritidis) contained no extra bands and matched completely with their autoradiographs. It was concluded that contamination of washed mycoplasma cells by serum proteins is mainly due to co-precipitation of aggregated serum proteins together with cells during centrifugation rather than to adsorption of serum proteins on the cell surface. Images Plate 1 PMID:10333

  18. Clinical Characteristics Associated with Mycoplasma genitalium among Female Sex Workers in Nairobi, Kenya

    PubMed Central

    Gomih-Alakija, Ayodele; Ting, Jie; Mugo, Nelly; Kwatampora, Jessie; Getman, Damon; Chitwa, Michael; Patel, Suha; Gokhale, Mugdha; Kimani, Joshua; Behets, Frieda S.

    2014-01-01

    The prevalence of Mycoplasma genitalium is high in vulnerable populations of women in low-resource settings. However, the epidemiology of infection in these populations is not well established. To determine the prevalence of Mycoplasma genitalium and its association with cervical cytology and other correlates, we recruited 350 female sex workers (FSW) who were 18 to 50 years old in Nairobi, Kenya, for a cross-sectional study. A questionnaire was administered at baseline to obtain information on sociodemographics and sexual behaviors. Women underwent a pelvic exam, during which a physician collected cervical-exfoliation samples for conventional cytology and sexually transmitted infection (STI) testing. Samples were tested for M. genitalium and other STI organisms (Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis) and the E6/E7 mRNA of human papillomavirus (HPV) by Aptima nucleic amplification assays. The prevalence of M. genitalium was 12.9%. FSW who engaged in sexual intercourse during menses were less likely to have M. genitalium infection than those who did not (odds ratio [OR], 0.3; 95% confidence interval [95% CI], 0.1, 0.9). M. genitalium was also less prevalent among FSW who had worked in prostitution for >5 years (6.2%) than among those who had worked for <3 years (17.6%) (OR, 0.3; 95% CI, 0.1, 0.8). FSW who reported more frequent condom use were more likely to be infected with M. genitalium than those who reported less frequent use (OR, 3.8; 95% CI, 1.2, 11.6). These correlates differ from those found in M. genitalium studies conducted with FSW from West Africa and China. Further longitudinal analyses assessing associations with persistent M. genitalium infection are needed. PMID:25100823

  19. Clinical characteristics associated with Mycoplasma genitalium among female sex workers in Nairobi, Kenya.

    PubMed

    Gomih-Alakija, Ayodele; Ting, Jie; Mugo, Nelly; Kwatampora, Jessie; Getman, Damon; Chitwa, Michael; Patel, Suha; Gokhale, Mugdha; Kimani, Joshua; Behets, Frieda S; Smith, Jennifer S

    2014-10-01

    The prevalence of Mycoplasma genitalium is high in vulnerable populations of women in low-resource settings. However, the epidemiology of infection in these populations is not well established. To determine the prevalence of Mycoplasma genitalium and its association with cervical cytology and other correlates, we recruited 350 female sex workers (FSW) who were 18 to 50 years old in Nairobi, Kenya, for a cross-sectional study. A questionnaire was administered at baseline to obtain information on sociodemographics and sexual behaviors. Women underwent a pelvic exam, during which a physician collected cervical-exfoliation samples for conventional cytology and sexually transmitted infection (STI) testing. Samples were tested for M. genitalium and other STI organisms (Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis) and the E6/E7 mRNA of human papillomavirus (HPV) by Aptima nucleic amplification assays. The prevalence of M. genitalium was 12.9%. FSW who engaged in sexual intercourse during menses were less likely to have M. genitalium infection than those who did not (odds ratio [OR], 0.3; 95% confidence interval [95% CI], 0.1, 0.9). M. genitalium was also less prevalent among FSW who had worked in prostitution for >5 years (6.2%) than among those who had worked for <3 years (17.6%) (OR, 0.3; 95% CI, 0.1, 0.8). FSW who reported more frequent condom use were more likely to be infected with M. genitalium than those who reported less frequent use (OR, 3.8; 95% CI, 1.2, 11.6). These correlates differ from those found in M. genitalium studies conducted with FSW from West Africa and China. Further longitudinal analyses assessing associations with persistent M. genitalium infection are needed. PMID:25100823

  20. Dissociation between learning and memory impairment and other sickness behaviours during simulated Mycoplasma infection in rats.

    PubMed

    Swanepoel, Tanya; Harvey, Brian H; Harden, Lois M; Laburn, Helen P; Mitchell, Duncan

    2011-11-01

    To investigate potential consequences for learning and memory, we have simulated the effects of Mycoplasma infection, in rats, by administering fibroblast-stimulating lipopepide-1 (FSL-1), a pyrogenic moiety of Mycoplasma salivarium. We measured the effects on body temperature, cage activity, food intake, and on spatial learning and memory in a Morris Water Maze. Male Sprague-Dawley rats had radio transponders implanted to measure abdominal temperature and cage activity. After recovery, rats were assigned randomly to receive intraperitoneal (I.P.) injections of FSL-1 (500 or 1000 ?g kg(-1) in 1 ml kg(-1) phosphate-buffered saline; PBS) or vehicle (PBS, 1 ml kg(-1)). Body mass and food intake were measured daily. Training in the Maze commenced 18 h after injections and continued daily for four days. Spatial memory was assessed on the fifth day. In other rats, we measured concentrations of brain pro-inflammatory cytokines, interleukin (IL)-1? and IL-6, at 3 and 18 h after injections. FSL-1 administration induced a dose-dependent fever (?1°C) for two days, lethargy (?78%) for four days, anorexia (?65%) for three days and body mass stunting (?6%) for at least four days. Eighteen hours after FSL-1 administration, when concentrations of IL-1?, but not that of IL-6, were elevated in both the hypothalamus and the hippocampus, and when rats were febrile, lethargic and anorexic, learning in the Maze was unaffected. There also was no memory impairment. Our results support emerging evidence that impaired learning and memory is not inevitable during simulated infection. PMID:21635947

  1. Dynamics of an Infectious Keratoconjunctivitis Outbreak by Mycoplasma conjunctivae on Pyrenean Chamois Rupicapra p. pyrenaica

    PubMed Central

    de la Fe, Christian; Revilla, Miguel; Prada, Carlos; Martínez-Durán, David; Gómez-Martín, Ángel; Fernández-Arberas, Olatz; Amores, Joaquín; Contreras, Antonio; García-Serrano, Alicia; de Luco, Daniel Fernández

    2013-01-01

    Between 2006 and 2008, an outbreak of Infectious Keratoconjunctivitis (IKC) affected Pyrenean chamois Rupicapra p. pyrenaica, an endemic subspecies of mountain ungulate that lives in the Pyrenees. The study focused on 14 mountain massifs (180,000 ha) where the species’ population is stable. Cases of IKC were detected in ten of the massifs and, in five of them, mortality was substantial. The outbreak spread quickly from the first location detected, with two peaks in mortality that affected one (2007) and three (2008) massifs. In the latter, the peak was seasonal (spring to autumn) and, in the former, the outbreak persisted through winter. To identify the outbreak’s aetiology, we examined 105 Pyrenean chamois clinically affected with IKC. TaqMan rt-PCR identified Mycoplasma conjunctivae in 93 (88.5%) of the chamois. Another rt-PCR detected Chlamydophila spp. in 14 of chamois, and 12 of those had mixed infections with mycoplasmas. In the period 2000–2007, the chamois population increased slightly (? 1.026) but decreased significantly during the IKC outbreak (? 0.8, 2007–2008; ? 0.85, 2008–2009) before increasing significantly after the outbreak (? 1.1, 2009–2010). Sex-biased mortality shifted the adult sex ratio toward males (from 0.6 to 0.7 males per female) and reduced productivity slightly. Hunting was practically banned in the massifs where chamois experienced significant mortality and allowed again after the outbreak ended. Long-term monitoring of wild populations provides a basis for understanding the impacts of disease outbreaks and improves management decisions, particularly when species are subject to extractive exploitation. PMID:23637923

  2. Syndrome de Miller Fisher avec anticorps anti GQ1b négatif au cours d'une pneumonie à Mycoplasma pneumoniae

    PubMed Central

    Sini, Victor; Tegueu, Calixte Kuate; Nguefack, Séraphin; Boone, Mathieu; Roos-Weil, Richard

    2013-01-01

    Le Syndrome de Miller Fisher est caractérisé par l'association d'une ophtalmoplégie, d'une ataxie et d'une aréflexie ostéo-tendineuse. Une infection virale est le plus souvent retrouvée dans les jours ou semaines qui précèdent la symptomatologie. Nous rapportons un cas de syndrome de Miller Fisher survenu chez une femme de 75 ans, et ce au décours d'une infection pulmonaire à Mycoplasma pneumoniae. Les sérologies virales habituelles étaient négatives. Les anticorps anti GQ1b étaient absents. Il n'y avait pas de lésion du tronc cérébral à l'imagerie par résonnance magnétique. L’évolution clinique était favorable après perfusion d'immunoglobulines humaines polyvalentes et des macrolides en comprimés. La sérologie Mycoplasma pneumoniae doit être systématiquement recherchée dans le bilan du syndrome de Miller Fisher. PMID:24255728

  3. Variable lipoprotein haemagglutinin (vlhA) gene sequence typing of mainly Dutch Mycoplasma synoviae isolates: comparison with vlhA sequences from Genbank and with amplified fragment length polymorphism analysis.

    PubMed

    Dijkman, R; Feberwee, A; Landman, W J M

    2014-01-01

    Molecular typing techniques with sufficient discriminatory power are required to better understand the transmission of Mycoplasma synoviae, a poultry pathogen with increasing clinical and economic relevance. A promising molecular technique is polymerase chain reaction and subsequent sequencing based on the conserved 5' region of the M. synoviae variable lipoprotein and haemagglutinin (vlhA) gene. This technique was used for genotyping 27 mainly Dutch M. synoviae isolates from different organs of various categories of poultry housed on different farms and collected during a period of 10 years. The obtained vlhA sequences were compared with those of M. synoviae strains from Genbank and data obtained by amplified fragment length polymorphism (AFLP). Grouping based on 100% similarity revealed nine genotypes. Some isolates had identical vlhA gene sequences although they originated from different geographical areas, different years and organs. AFLP analysis results largely confirmed the results obtained by vlhA sequence typing. Our findings raise concern regarding the discriminatory power of these techniques for its use in molecular epidemiology of Dutch M. synoviae isolates and for the differentiation between M. synoviae vaccine strains and field isolates, and indicate that molecular typing based on additional markers should be considered. PMID:25189763

  4. The effect of thiol-active compounds and sterols on the membrane-associated hemolysin of Mycoplasma pulmonis

    Microsoft Academic Search

    Karalee J. Jarvill-Taylor; F. Chris Minion

    1995-01-01

    Previous studies had shown that Mycoplasma pulmonis contained a bovine serum albumin-dependent, membrane-associated hemolysin. Biochemical analyses were performed to further characterize this activity. The membrane-associated hemolytic activity could be activated by dithiothreitol and ?-mercaptoethanol, and inactivated by oxidizing compounds, a sulfhydryl inhibitor and heat treatment. Cholesterol and other sterols were inhibitory in a stereo-specific manner, but they did not interfere

  5. Effect of vaccination against Mycoplasma hyopneumoniae in pig herds with an all-in\\/all-out production system

    Microsoft Academic Search

    D Maes; H Deluyker; M Verdonck; F Castryck; C Miry; B Vrijens; W Verbeke; J Viaene; A de Kruif

    1999-01-01

    A multi-site field study was conducted to evaluate an inactivated Mycoplasma hyopneumoniae (Mh) vaccine in 14 pig herds infected by Mh and practising an all-in\\/all-out production system. In each herd, a vaccinated and control group of 250 pigs each were compared during the growing\\/finishing period with respect to performance parameters (major variables) and by means of clinical, serological and pathological

  6. Mycoplasma pneumoniae Community Acquired Respiratory Distress Syndrome toxin expression reveals growth phase and infection-dependent regulation

    PubMed Central

    Kannan, T R; Musatovova, Oxana; Balasubramanian, Sowmya; Cagle, Marianna; Jordan, Jarrat L; Krunkosky, Thomas M; Davis, Alan; Hardy, Robert D; Baseman, Joel B

    2010-01-01

    Mycoplasma pneumoniae causes acute and chronic respiratory infections, including tracheobronchitis and community acquired pneumonia, and is linked to asthma and an array of extra-pulmonary disorders. Recently, we identified an ADP-ribosylating and vacuolating toxin of M. pneumoniae, designated Community Acquired Respiratory Distress Syndrome (CARDS) toxin. In this study we analysed CARDS toxin gene (annotated mpn372) transcription and identified its promoter. We also compared CARDS toxin mRNA and protein profiles in M. pneumoniae during distinct in vitro growth phases. CARDS toxin mRNA expression was maximal, but at low levels, during early exponential growth and declined sharply during mid-to-late log growth phases, which was in direct contrast to other mycoplasma genes examined. Between 7% and 10% of CARDS toxin was localized to the mycoplasma membrane at mid-exponential growth, which was reinforced by immunogold electron microscopy. No CARDS toxin was released into the medium. Upon M. pneumoniae infection of mammalian cells, increased expression of CARDS toxin mRNA was observed when compared with SP-4 broth-grown cultures. Further, confocal immunofluorescence microscopy revealed that M. pneumoniae readily expressed CARDS toxin during infection of differentiated normal human bronchial epithelial cells. Analysis of M. pneumoniae-infected mouse lung tissue revealed high expression of CARDS toxin per mycoplasma cell when compared with M. pneumoniae cells grown in SP-4 medium alone. Taken together, these studies indicate that CARDS toxin expression is carefully controlled by environmental cues that influence its transcription and translation. Further, the acceleration of CARDS toxin synthesis and accumulation in vivo is consistent with its role as a bona fide virulence determinant. PMID:20199607

  7. Killing rate curve and combination effects of surfactin C produced from Bacillus subtilis complex BC1212 against pathogenic Mycoplasma hyopneumoniae

    Microsoft Academic Search

    M. H. Hwang; M. H. Kim; E. Gebru; B. Y. Jung; S. P. Lee; S. C. Park

    2008-01-01

    This study evaluated the killing rate as well as the antimycoplasmal effect of surfactins isolated from Bacillus subtilis complex BC1212, either alone or in combination with various antibacterials against Mycoplasma hyopneumoniae. Prior to the killing rate and the combination effect studies of surfactins, the minimal inhibitory concentrations (MICs)\\u000a of various antibacterials and surfactins (consisting of surfactins A, B, C, and

  8. Elongated Versions of Vlp Surface Lipoproteins Protect Mycoplasma hyorhinisEscape Variants from Growth Inhibiting Host Antibodies

    Microsoft Academic Search

    CHRISTINE CITTI; MARY F. KIM; ANDKIM S. WISE

    1997-01-01

    Variation in Vlp surface proteins ofMycoplasma hyorhiniswas evaluated in terms of its role in determining susceptibility of organisms to growth inhibition by host antibodies (Abs). High-frequency switching of Vlp surface lipoproteins has been studied in isogenic lineages ofM. hyorhinisSK76. In these lineages, the products ofthreegenes,vlpA,vlpB,andvlpC,aresubjecttophaseandsizevariationinvitro,whichoccurthroughdistinct mutator elements that independently govern the expression of eachvlpgene (promoter mutations) or the size of

  9. Characterization of Cleavage Events in the Multifunctional Cilium Adhesin Mhp684 (P146) Reveals a Mechanism by Which Mycoplasma hyopneumoniae Regulates Surface Topography

    PubMed Central

    Bogema, Daniel R.; Deutscher, Ania T.; Woolley, Lauren K.; Seymour, Lisa M.; Raymond, Benjamin B. A.; Tacchi, Jessica L.; Padula, Matthew P.; Dixon, Nicholas E.; Minion, F. Chris; Jenkins, Cheryl; Walker, Mark J.; Djordjevic, Steven P.

    2012-01-01

    ABSTRACT Mycoplasma hyopneumoniae causes enormous economic losses to swine production worldwide by colonizing the ciliated epithelium in the porcine respiratory tract, resulting in widespread damage to the mucociliary escalator, prolonged inflammation, reduced weight gain, and secondary infections. Protein Mhp684 (P146) comprises 1,317 amino acids, and while the N-terminal 400 residues display significant sequence identity to the archetype cilium adhesin P97, the remainder of the molecule is novel and displays unusual motifs. Proteome analysis shows that P146 preprotein is endogenously cleaved into three major fragments identified here as P50P146, P40P146, and P85P146 that reside on the cell surface. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) identified a semitryptic peptide that delineated a major cleavage site in Mhp684. Cleavage occurred at the phenylalanine residue within sequence 672ATEF?QQ677, consistent with a cleavage motif resembling S/T-X-F?X-D/E recently identified in Mhp683 and other P97/P102 family members. Biotinylated surface proteins recovered by avidin chromatography and separated by two-dimensional gel electrophoresis (2-D GE) showed that more-extensive endoproteolytic cleavage of P146 occurs. Recombinant fragments F1P146-F3P146 that mimic P50P146, P40P146, and P85P146 were constructed and shown to bind porcine epithelial cilia and biotinylated heparin with physiologically relevant affinity. Recombinant versions of F3P146 generated from M. hyopneumoniae strain J and 232 sequences strongly bind porcine plasminogen, and the removal of their respective C-terminal lysine and arginine residues significantly reduces this interaction. These data reveal that P146 is an extensively processed, multifunctional adhesin of M. hyopneumoniae. Extensive cleavage coupled with variable cleavage efficiency provides a mechanism by which M. hyopneumoniae regulates protein topography. PMID:22493032

  10. Allelic polymorphisms at the H-2A and HLA-DQ loci influence the response of murine lymphocytes to the Mycoplasma arthritidis superantigen MAM.

    PubMed Central

    Cole, B C; Sawitzke, A D; Ahmed, E A; Atkin, C L; David, C S

    1997-01-01

    Mycoplasma arthritidis, an agent of rodent arthritis, produces a potent superantigen (SAg), MAM. Previous work established that MAM is presented to T cells by murine H-2E or the homologous human HLA-DR molecules and that lymphocytes lacking a functional H-2E molecule fail to respond to MAM. Recently, more potent and purified preparations of MAM of known protein content have become available. This enabled us to more effectively compare the response of MAM with that of other SAgs by using lymphocytes from mice whose cells express different H-2A and HLA-DQ molecules. Here we demonstrate that cells from some H-2E-negative mouse strains respond to higher concentrations of MAM. By use of inbred, congenic, and recombinant mice, we show that these differences are, in fact, exercised at the level of the major histocompatibility complex (MHC) and that allelic polymorphisms at H-2A influence reactivity to MAM. In addition, polymorphisms at HLA-DQ, the human homolog of H-2A, also influence responsiveness to MAM. Cells expressing DQw6 (HLA-DQA1*0103 and DQBI*0601 chains) gave much higher responses to MAM than did cells expressing DQw8 (DQA1*0301 and DQB1*0302 chains). In fact, responses of lymphocytes expressing DQB1*0601 chains homozygously were as high as those observed for cells expressing a functional H-2E molecule. Murine lymphocytes responded less well to staphylococcal enterotoxin B (SEB) and SEA, but mouse cells expressing human MHC molecules gave much higher responses. The patterns of reactivity observed with cells expressing the various murine and human alleles differed for MAM, SEB, and SEA, suggesting that each of these SAgs interacts with different regions or residues on MHC molecules. It has been hypothesized that SAgs might play a role in susceptibility to autoimmune disease. Allelic polymorphisms at MHC loci might therefore influence susceptibility to autoimmune disease by affecting immunoreactivity to specific superantigens. PMID:9317026

  11. Simultaneous Identification of Potential Pathogenicity Factors of Mycoplasma agalactiae in the Natural Ovine Host by Negative Selection.

    PubMed

    Hegde, Shivanand; Hegde, Shrilakshmi; Zimmermann, Martina; Flöck, Martina; Spergser, Joachim; Rosengarten, Renate; Chopra-Dewasthaly, Rohini

    2015-07-01

    Mycoplasmas possess complex pathogenicity determinants that are largely unknown at the molecular level. Mycoplasma agalactiae serves as a useful model to study the molecular basis of mycoplasma pathogenicity. The generation and in vivo screening of a transposon mutant library of M. agalactiae were employed to unravel its host colonization factors. Tn4001mod mutants were sequenced using a novel sequencing method, and functionally heterogeneous pools containing 15 to 19 selected mutants were screened simultaneously through two successive cycles of sheep intramammary infections. A PCR-based negative selection method was employed to identify mutants that failed to colonize the udders and draining lymph nodes in the animals. A total of 14 different mutants found to be absent from ?95% of samples were identified and subsequently verified via a second round of stringent confirmatory screening where 100% absence was considered attenuation. Using this criterion, seven mutants with insertions in genes MAG1050, MAG2540, MAG3390, uhpT, eutD, adhT, and MAG4460 were not recovered from any of the infected animals. Among the attenuated mutants, many contain disruptions in hypothetical genes, implying their previously unknown role in M. agalactiae pathogenicity. These data indicate the putative role of functionally different genes, including hypothetical ones, in the pathogenesis of M. agalactiae. Defining the precise functions of the identified genes is anticipated to increase our understanding of M. agalactiae infections and to develop successful intervention strategies against it. PMID:25916984

  12. High temperature strain gage apparent strain compensation

    NASA Technical Reports Server (NTRS)

    Holmes, Harlan K.; Moore, T. C., Sr.

    1992-01-01

    Once an installed strain gage is connected to a strain indicating device and the instrument is balanced, a subsequent change in temperature of the gage installation will generally produce a resistance change in the gage. This purely temperature-induced resistance will be registered by the indicating device as a strain and is referred to as 'apparent strain' to distinguish it from strain due to applied stress. One desirable technique for apparent strain compensation is to employ two identical gages with identical mounting procedures which are connected with a 'half bridge' configuration where gages see the same thermal environment but only one experiences a mechanical strain input. Their connection in adjacent arms of the bridge will then balance the thermally induced apparent strains and, in principle, only the mechanical strain remains. Two approaches that implement this technique are discussed.

  13. Mouse Repository Strain Details

    Cancer.gov

    Available Strain Details Order Form for Cryoarchived Strains   Strain Number: 01XE7  Common Strain Name: Villin-Cre, vil-Cre Fo20  Strain Nomenclature: B6.Cg-Tg(Vil-cre)20Sy/Nci  Release Category (Required for MTA form): B1 , D Sample MTA for this

  14. Mouse Repository Strain Details

    Cancer.gov

    Available Strain Details Order Form for Live Mice   Strain Number: 01XBL  Common Strain Name: Myf6-ires-cre knock-in  Strain Nomenclature: B6;129-Myf6tm2(cre)Mrc/Nci  Release Category (Required for MTA form): C1 , D Sample MTA for this strain Animal

  15. Muscle strain (image)

    MedlinePLUS

    A muscle strain is the stretching or tearing of muscle fibers. A muscle strain can be caused by sports, exercise, a ... something that is too heavy. Symptoms of a muscle strain include pain, tightness, swelling, tenderness, and the ...

  16. Role of Mycoplasma penetrans Endonuclease P40 as a Potential Pathogenic Determinant

    PubMed Central

    Bendjennat, Mourad; Blanchard, Alain; Loutfi, Mohammed; Montagnier, Luc; Bahraoui, Elmostafa

    1999-01-01

    Recently, we reported the purification to homogeneity and characterization of Ca2+- and Mg2+-dependent endonuclease P40 produced by Mycoplasma penetrans (M. Bendjennat, A. Blanchard, M. Loutfi, L. Montagnier, and E. Bahraoui, J. Bacteriol. 179; 2210–2220, 1997), a mycoplasma which was isolated for the first time from the urine of human immunodeficiency virus-infected patients. To evaluate how this nuclease could interact with host cells, we tested its effect on CEM and Molt-4 lymphocytic cell lines and on peripheral blood mononuclear cells. We observed that 10?7 to 10?9 M P40 is able to mediate a cytotoxic effect. We found that 100% of cells were killed after 24 h of incubation with 10?7 M P40 while only 40% cytotoxicity was obtained after 72 h of incubation with 10?9 M P40. Phase-contrast microscopy observations of P40-treated cells revealed morphological changes, including pronounced blebbing of the plasma membrane and cytoplasmic shrinkage characteristic of programmed cell death, which is in agreement with the internucleosomal fragmentation of P40-treated cell DNA as shown by agarose gel electrophoresis. We showed that 125I-radiolabeled or fluorescein isothiocyanate-labeled P40 was able to bind specifically in a dose-dependent manner to the cell membrane of CEM cells, which suggested that the cytotoxicity of P40 endonuclease was mediated by its interaction with the cell surface receptor(s). The concentration of unlabeled P40 required to inhibit by 50% the formation of 125I-P40-CEM complexes was about 3 × 10?9 M, indicating a high-affinity interaction. Both P40 interaction and cytotoxicity are Ca2+ dependent. Our results suggest that the cytotoxicity of M. penetrans observed in vitro is mediated at least partially by secreted P40, which, after interaction with host cells, can induce an apoptosis-like death. These results strongly suggest a major role of mycoplasmal nucleases as potential pathogenic determinants. PMID:10456886

  17. Role of Mycoplasma penetrans endonuclease P40 as a potential pathogenic determinant.

    PubMed

    Bendjennat, M; Blanchard, A; Loutfi, M; Montagnier, L; Bahraoui, E

    1999-09-01

    Recently, we reported the purification to homogeneity and characterization of Ca(2+)- and Mg(2+)-dependent endonuclease P40 produced by Mycoplasma penetrans (M. Bendjennat, A. Blanchard, M. Loutfi, L. Montagnier, and E. Bahraoui, J. Bacteriol. 179; 2210-2220, 1997), a mycoplasma which was isolated for the first time from the urine of human immunodeficiency virus-infected patients. To evaluate how this nuclease could interact with host cells, we tested its effect on CEM and Molt-4 lymphocytic cell lines and on peripheral blood mononuclear cells. We observed that 10(-7) to 10(-9) M P40 is able to mediate a cytotoxic effect. We found that 100% of cells were killed after 24 h of incubation with 10(-7) M P40 while only 40% cytotoxicity was obtained after 72 h of incubation with 10(-9) M P40. Phase-contrast microscopy observations of P40-treated cells revealed morphological changes, including pronounced blebbing of the plasma membrane and cytoplasmic shrinkage characteristic of programmed cell death, which is in agreement with the internucleosomal fragmentation of P40-treated cell DNA as shown by agarose gel electrophoresis. We showed that (125)I-radiolabeled or fluorescein isothiocyanate-labeled P40 was able to bind specifically in a dose-dependent manner to the cell membrane of CEM cells, which suggested that the cytotoxicity of P40 endonuclease was mediated by its interaction with the cell surface receptor(s). The concentration of unlabeled P40 required to inhibit by 50% the formation of (125)I-P40-CEM complexes was about 3 x 10(-9) M, indicating a high-affinity interaction. Both P40 interaction and cytotoxicity are Ca(2+) dependent. Our results suggest that the cytotoxicity of M. penetrans observed in vitro is mediated at least partially by secreted P40, which, after interaction with host cells, can induce an apoptosis-like death. These results strongly suggest a major role of mycoplasmal nucleases as potential pathogenic determinants. PMID:10456886

  18. A field evaluation of two vaccines against Mycoplasma hyopneumoniae infection in pigs

    PubMed Central

    2014-01-01

    Background A field trial was carried out with two Mycoplasma hyopneumoniae vaccines in order to investigate the benefit of vaccination under field conditions in modern Danish pig production facilities with pigs being positive for M. hyopneumoniae. The M. hyopneumoniae infection of the herd was confirmed through blood samples that were positive for antibodies against M. hyopneumoniae combined with gross lesions of the lungs related to M. hyopneumoniae at slaughter and detection of M. hyopneumoniae by polymerace chain reaction in these lesions. Results A total of 2,256 pigs from two herds were randomly divided into three groups. Group 1 received 2 mL ThoroVAX®VET, Group 2 received 1 mL Ingelvac®MycoFLEX, and Group 3 was a non-vaccinated control group. The vaccination was performed by a person who was not involved in the rest of the trial and vaccination status thereby blinded to the evaluators. The prevalence of lung lesions related to M. hyopneumoniae were significantly lower for pigs vaccinated with ThoroVAX®VET but not for pigs vaccinated with Ingelvac®MycoFLEX®, when compared to non-vaccinated pigs. There was no significant effect of vaccination on growth rate, antibiotic consumption or mortality. Conclusion This trial demonstrated that vaccination with Thoro®VAX VET was effective in reducing the prevalence of lung lesion in pig units infected with M. hyopneumoniae. PMID:24739629

  19. Dissecting the energy metabolism in Mycoplasma pneumoniae through genome-scale metabolic modeling

    PubMed Central

    Wodke, Judith A H; Pucha?ka, Jacek; Lluch-Senar, Maria; Marcos, Josep; Yus, Eva; Godinho, Miguel; Gutiérrez-Gallego, Ricardo; dos Santos, Vitor A P Martins; Serrano, Luis; Klipp, Edda; Maier, Tobias

    2013-01-01

    Mycoplasma pneumoniae, a threatening pathogen with a minimal genome, is a model organism for bacterial systems biology for which substantial experimental information is available. With the goal of understanding the complex interactions underlying its metabolism, we analyzed and characterized the metabolic network of M. pneumoniae in great detail, integrating data from different omics analyses under a range of conditions into a constraint-based model backbone. Iterating model predictions, hypothesis generation, experimental testing, and model refinement, we accurately curated the network and quantitatively explored the energy metabolism. In contrast to other bacteria, M. pneumoniae uses most of its energy for maintenance tasks instead of growth. We show that in highly linear networks the prediction of flux distributions for different growth times allows analysis of time-dependent changes, albeit using a static model. By performing an in silico knock-out study as well as analyzing flux distributions in single and double mutant phenotypes, we demonstrated that the model accurately represents the metabolism of M. pneumoniae. The experimentally validated model provides a solid basis for understanding its metabolic regulatory mechanisms. PMID:23549481

  20. [Parainfectious polyneuropathy and miller-fisher-syndrome in combination with anemia in Mycoplasma pneumoniae infection].

    PubMed

    Loewenbrück, K F; Pütz, V; Schäfer, J; Reichmann, H; Storch, A

    2008-06-01

    Neurological complications are among the most common in Mycoplasma pneumoniae (M.p.) infection and appear in up to 4,8 % of M.p.-infected patients . Whereas direct intracerebral infection can occur and often puts patients into critical, intensive care-requiring conditions, different forms of parainfectious autoimmune Guillain-Barré-syndrome (GBS) and Miller-Fisher syndrome (MFS) make up the most frequent neurological complications. M.p.-associated alterations of the blood count are common, and sometimes can take a dramatical course. We here report a 64-year-old female and a 53-year-old male patient who both suffered from severe parainfectious autoimmune neurological syndromes in combination with different forms of anemia, in the case of the first patient to a transfusion-requiring degree. With this report we would like to stress the importance to include M.p. in the diagnostic procedures in the case of inflammatory neurological syndromes, especially when combined with alterations of the blood count. PMID:18512187

  1. Mycoplasma fermentans glycolipid-antigen as a pathogen of rheumatoid arthritis

    SciTech Connect

    Kawahito, Yutaka [Inflammation and Immunology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, 465 Kajii-ku, Kawaramachi-Hirokoji Kamigyo-ku, Kyoto-shi, Kyoto 602-0841 (Japan); Ichinose, Sizuko [Laboratory for Electron Microscopy, Tokyo Medical and Dental University, 5-45, Yushima 1-chome, Bunkyo-ku, Tokyo 113-8519 (Japan); Sano, Hajime [Arthiritis and Rheumatism Branch, Department of Internal Medicine, Hyogo Medical College of Medicine, 1-1, Nishinomiya, Mukogawa-cho, Hyogo 663-8501 (Japan); Tsubouchi, Yasunori; Kohno, Masataka; Yoshikawa, Toshikazu [Inflammation and Immunology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, 465 Kajii-ku, Kawaramachi-Hirokoji Kamigyo-ku, Kyoto-shi, Kyoto 602-0841 (Japan); Tokunaga, Daisaku; Hojo, Tatsuya [Department of Orthopedics, Kyoto Prefectural University of Medicine, 465 Kajii-ku, Kawaramachi-Hirokoji Kamigyo-ku, Kyoto, Kyoto 602-0841 (Japan); Harasawa, Ryo [Department of Veterinary Medicine, Faculty of Agriculture, Iwate University, 3-18-8, Ueda, Morioka, Iwate 020-8550 (Japan); Nakano, Teruaki [Department of Rheumatology and Internal medicine, OUR LADY OF SNOW Medical Juridical Corporation ST. MARY'S HOSPITAL, 422 Tsubukuhon-machi Kurume City, Fukuoka 830-8543 (Japan); Matsuda, Kazuhiro [Department of Research and Developments, M Bio Technology Inc., Koto-ku, TIME 24 Building 10F, Aomi 2-45, Koto-ku, Tokyo 135-8073 (Japan)], E-mail: matsuda-k@mbiotech.org

    2008-05-02

    Mycoplasma fermentans has been suspected as one of the causative pathogenic microorganisms of rheumatoid arthritis (RA) however, the pathogenic mechanism is still unclear. We, previously, reported that glycolipid-antigens (GGPL-I and III) are the major antigens of M. fermentans. Monoclonal antibody against the GGPL-III could detect the existence of the GGPL-III antigens in synovial tissues from RA patients. GGPL-III antigens were detected in 38.1% (32/84) of RA patient's tissues, but not in osteoarthritis (OA) and normal synovial tissues. Immunoelectron microscopy revealed that a part of GGPL-III antigens are located at endoplasmic reticulum. GGPL-III significantly induced TNF-{alpha} and IL-6 production from peripheral blood mononulear cells, and also proliferation of synovial fibroblasts. Further study is necessary to prove that M. fermentans is a causative microorganism of RA; however, the new mechanisms of disease pathogenesis provides hope for the development of effective and safe immunotherapeutic strategies based on the lipid-antigen, GGPL-III, in the near future.

  2. Transcription Analysis of the Porcine Alveolar Macrophage Response to Mycoplasma hyopneumoniae

    PubMed Central

    Bin, Li; Luping, Du; Bing, Sun; Zhengyu, Yu; Maojun, Liu; Zhixin, Feng; Yanna, Wei; Haiyan, Wang; Guoqing, Shao; Kongwang, He

    2014-01-01

    Mycoplasma hyopneumoniae is considered the major causative agent of porcine respiratory disease complex, occurs worldwide and causes major economic losses to the pig industry. To gain more insights into the pathogenesis of this organism, the high throughput cDNA microarray assays were employed to evaluate host responses of porcine alveolar macrophages to M. hyopneumoniae infection. A total of 1033 and 1235 differentially expressed genes were identified in porcine alveolar macrophages in responses to exposure to M. hyopneumoniae at 6 and 15 hours post infection, respectively. The differentially expressed genes were involved in many vital functional classes, including inflammatory response, immune response, apoptosis, cell adhesion, defense response, signal transduction, protein folding, protein ubiquitination and so on. The pathway analysis demonstrated that the most significant pathways were the chemokine signaling pathway, Toll-like receptor signaling pathway, RIG-I-like receptor signaling pathway, nucleotide-binding oligomerization domains (Nod)-like receptor signaling pathway and apoptosis signaling pathway. The reliability of the data obtained from the microarray was verified by performing quantitative real-time PCR. The expression kinetics of chemokines was further analyzed. The present study is the first to document the response of porcine alveolar macrophages to M. hyopneumoniae infection. The data further developed our understanding of the molecular pathogenesis of M. hyopneumoniae. PMID:25098731

  3. Mycoplasma pneumoniae outbreak in a long-term care facility--Nebraska, 2014.

    PubMed

    Hastings, Deborah L; Harrington, Kari J; Kutty, Preeta K; Rayman, Rebecca J; Spindola, Dana; Diaz, Maureen H; Thurman, Kathleen A; Winchell, Jonas M; Safranek, Thomas J

    2015-03-27

    On June 20, 2014, a Nebraska long-term care facility notified the East Central District Health Department (ECDHD) and Nebraska Department of Health and Human Services (NDHHS) of an outbreak of respiratory illness characterized by cough and fever in 22 residents and resulting in four deaths during the preceding 2 weeks. To determine the etiologic agent, identify additional cases, and implement control measures, Nebraska and CDC investigators evaluated the facility's infection prevention measures and collected nasopharyngeal (NP) and oropharyngeal (OP) swabs or autopsy specimens from patients for real-time polymerase chain reaction (PCR) testing at CDC. The facility was closed to new admissions until 1 month after the last case, droplet precautions were implemented, ill residents were isolated, and group activities were canceled. During the outbreak, a total of 55 persons experienced illnesses that met the case definition; 12 were hospitalized, and seven died. PCR detected Mycoplasma pneumoniae DNA in 40% of specimens. M. pneumoniae should be considered a possible cause of respiratory illness outbreaks in long-term care facilities. Morbidity and mortality from respiratory disease outbreaks at long-term care facilities might be minimized if facilities monitor for respiratory disease clusters, report outbreaks promptly, prioritize diagnostic testing in outbreak situations, and implement timely and strict infection control measures to halt transmission. PMID:25811678

  4. Necrotic pharyngitis associated with Mycoplasma bovis infections in American bison (Bison bison).

    PubMed

    Dyer, Neil; Register, Karen B; Miskimins, Dale; Newell, Teresa

    2013-03-01

    Mycoplasma bovis has emerged as a significant and costly infectious disease problem in bison, generally presenting as severe, caseonecrotic pneumonia. Three diagnostic cases in which M. bovis is associated with necrotic pharyngitis in bison are described in the current study. The bacterium was isolated from lesions of the pharynx or lung of 3 American bison (Bison bison), at 2 different locations in the upper Midwestern United States, with severe, necrotic pharyngeal abscesses. Chronic caseonecrotic inflammation typical of M. bovis infection in bovines was observed microscopically in the pharynxes of affected bison. A mixed population of bacteria was recovered from the pharyngeal lesions, and Trueperella pyogenes, a frequent secondary pathogen in ruminant respiratory disease, was consistently isolated from the affected animals. Distinctive histopathological features of the pharyngeal lesions favor causation by M. bovis, although a role for T. pyogenes in the clinical presentation cannot be excluded. Veterinarians and producers working with bison should be aware that M. bovis may be associated with pharyngitis in bison. PMID:23512925

  5. MyMpn: a database for the systems biology model organism Mycoplasma pneumoniae.

    PubMed

    Wodke, Judith A H; Alibés, Andreu; Cozzuto, Luca; Hermoso, Antonio; Yus, Eva; Lluch-Senar, Maria; Serrano, Luis; Roma, Guglielmo

    2015-01-01

    MyMpn (http://mympn.crg.eu) is an online resource devoted to studying the human pathogen Mycoplasma pneumoniae, a minimal bacterium causing lower respiratory tract infections. Due to its small size, its ability to grow in vitro, and the amount of data produced over the past decades, M. pneumoniae is an interesting model organisms for the development of systems biology approaches for unicellular organisms. Our database hosts a wealth of omics-scale datasets generated by hundreds of experimental and computational analyses. These include data obtained from gene expression profiling experiments, gene essentiality studies, protein abundance profiling, protein complex analysis, metabolic reactions and network modeling, cell growth experiments, comparative genomics and 3D tomography. In addition, the intuitive web interface provides access to several visualization and analysis tools as well as to different data search options. The availability and--even more relevant--the accessibility of properly structured and organized data are of up-most importance when aiming to understand the biology of an organism on a global scale. Therefore, MyMpn constitutes a unique and valuable new resource for the large systems biology and microbiology community. PMID:25378328

  6. Global multilocus sequence typing analysis of Mycoplasma bovis isolates reveals two main population clusters.

    PubMed

    Rosales, R S; Churchward, C P; Schnee, C; Sachse, K; Lysnyansky, I; Catania, S; Iob, L; Ayling, R D; Nicholas, R A J

    2015-03-01

    Mycoplasma bovis is a major bovine pathogen associated with bovine respiratory disease complex and is responsible for substantial economic losses worldwide. M. bovis is also associated with other clinical presentations in cattle, including mastitis, otitis, arthritis, and reproductive disorders. To gain a better understanding of the genetic diversity of this pathogen, a multilocus sequence typing (MLST) scheme was developed and applied to the characterization of 137 M. bovis isolates from diverse geographical origins, obtained from healthy or clinically infected cattle. After in silico analysis, a final set of 7 housekeeping genes was selected (dnaA, metS, recA, tufA, atpA, rpoD, and tkt). MLST analysis demonstrated the presence of 35 different sequence types (STs) distributed in two main clonal complexes (CCs), defined at the double-locus variant level, namely, CC1, which included most of the British and German isolates, and CC2, which was a more heterogeneous and geographically distant group of isolates, including European, Asian, and Australian samples. Index of association analysis confirmed the clonal nature of the investigated M. bovis population, based on MLST data. This scheme has demonstrated high discriminatory power, with the analysis showing the presence of genetically distant and divergent clusters of isolates predominantly associated with geographical origins. PMID:25540400

  7. MHJ_0461 is a multifunctional leucine aminopeptidase on the surface of Mycoplasma hyopneumoniae

    PubMed Central

    Jarocki, Veronica M.; Santos, Jerran; Tacchi, Jessica L.; Raymond, Benjamin B. A.; Deutscher, Ania T.; Jenkins, Cheryl; Padula, Matthew P.; Djordjevic, Steven P.

    2015-01-01

    Aminopeptidases are part of the arsenal of virulence factors produced by bacterial pathogens that inactivate host immune peptides. Mycoplasma hyopneumoniae is a genome-reduced pathogen of swine that lacks the genetic repertoire to synthesize amino acids and relies on the host for availability of amino acids for growth. M. hyopneumoniae recruits plasmin(ogen) onto its cell surface via the P97 and P102 adhesins and the glutamyl aminopeptidase MHJ_0125. Plasmin plays an important role in regulating the inflammatory response in the lungs of pigs infected with M. hyopneumoniae. We show that recombinant MHJ_0461 (rMHJ_0461) functions as a leucine aminopeptidase (LAP) with broad substrate specificity for leucine, alanine, phenylalanine, methionine and arginine and that MHJ_0461 resides on the surface of M. hyopneumoniae. rMHJ_0461 also binds heparin, plasminogen and foreign DNA. Plasminogen bound to rMHJ_0461 was readily converted to plasmin in the presence of tPA. Computational modelling identified putative DNA and heparin-binding motifs on solvent-exposed sites around a large pore on the LAP hexamer. We conclude that MHJ_0461 is a LAP that moonlights as a multifunctional adhesin on the cell surface of M. hyopneumoniae. PMID:25589579

  8. P65 Truncation Impacts P30 Dynamics during Mycoplasma pneumoniae Gliding

    PubMed Central

    Hasselbring, Benjamin M.; Sheppard, Edward S.

    2012-01-01

    The cell wall-less prokaryote Mycoplasma pneumoniae is a major cause of community-acquired bronchitis and pneumonia in humans. Colonization is mediated largely by a differentiated terminal organelle, which is also the leading end in gliding motility. Cytadherence-associated proteins P30 and P65 appear to traffic concurrently to the distal end of developing terminal organelles. Here, truncation of P65 due to transposon insertion in the corresponding gene resulted in lower gliding velocity, reduced cytadherence, and decreased steady-state levels of several terminal organelle proteins, including P30. Utilizing fluorescent protein fusions, we followed terminal organelle development over time. New P30 foci appeared at nascent terminal organelles in P65 mutants, as in the wild type. However, with forward cell motility, P30 in the P65 mutants appeared to drag toward the trailing cell pole, where it was released, yielding a fluorescent trail to which truncated P65 colocalized. In contrast, P30 was only rarely observed at the trailing end of gliding wild-type cells. Complementation with the recombinant wild-type P65 allele by transposon delivery restored P65 levels and stabilized P30 localization to the terminal organelle. PMID:22544269

  9. P65 truncation impacts P30 dynamics during Mycoplasma pneumoniae gliding.

    PubMed

    Hasselbring, Benjamin M; Sheppard, Edward S; Krause, Duncan C

    2012-06-01

    The cell wall-less prokaryote Mycoplasma pneumoniae is a major cause of community-acquired bronchitis and pneumonia in humans. Colonization is mediated largely by a differentiated terminal organelle, which is also the leading end in gliding motility. Cytadherence-associated proteins P30 and P65 appear to traffic concurrently to the distal end of developing terminal organelles. Here, truncation of P65 due to transposon insertion in the corresponding gene resulted in lower gliding velocity, reduced cytadherence, and decreased steady-state levels of several terminal organelle proteins, including P30. Utilizing fluorescent protein fusions, we followed terminal organelle development over time. New P30 foci appeared at nascent terminal organelles in P65 mutants, as in the wild type. However, with forward cell motility, P30 in the P65 mutants appeared to drag toward the trailing cell pole, where it was released, yielding a fluorescent trail to which truncated P65 colocalized. In contrast, P30 was only rarely observed at the trailing end of gliding wild-type cells. Complementation with the recombinant wild-type P65 allele by transposon delivery restored P65 levels and stabilized P30 localization to the terminal organelle. PMID:22544269

  10. A genome-scale metabolic reconstruction of Mycoplasma genitalium, iPS189.

    PubMed

    Suthers, Patrick F; Dasika, Madhukar S; Kumar, Vinay Satish; Denisov, Gennady; Glass, John I; Maranas, Costas D

    2009-02-01

    With a genome size of approximately 580 kb and approximately 480 protein coding regions, Mycoplasma genitalium is one of the smallest known self-replicating organisms and, additionally, has extremely fastidious nutrient requirements. The reduced genomic content of M. genitalium has led researchers to suggest that the molecular assembly contained in this organism may be a close approximation to the minimal set of genes required for bacterial growth. Here, we introduce a systematic approach for the construction and curation of a genome-scale in silico metabolic model for M. genitalium. Key challenges included estimation of biomass composition, handling of enzymes with broad specificities, and the lack of a defined medium. Computational tools were subsequently employed to identify and resolve connectivity gaps in the model as well as growth prediction inconsistencies with gene essentiality experimental data. The curated model, M. genitalium iPS189 (262 reactions, 274 metabolites), is 87% accurate in recapitulating in vivo gene essentiality results for M. genitalium. Approaches and tools described herein provide a roadmap for the automated construction of in silico metabolic models of other organisms. PMID:19214212

  11. Mycoplasma genitalium and preterm delivery at an urban community health center†

    PubMed Central

    Averbach, Sarah H.; Hacker, Michele R.; Yiu, Timothy; Modest, Anna Merport; Dimitrakoff, Jordan; Ricciotti, Hope A.

    2013-01-01

    Objective To determine the prepartum prevalence of cervical Mycoplasma genitaliu colonization and evaluate prospectively whether colonization is associated with preterm delivery among women from a racial/ethnic minority background with a high risk of delivering a low birth weight newborn and a high prevalence of sexually transmitted infections. Methods In a prospective cohort study at an urban community health center in Roxbury, MA, USA, 100 women receiving routine prenatal care for singleton pregnancies were enrolled between August 2010 and December 2011. Endocervical samples were tested for M. genitalium, and delivery data were collected. Results The prevalence of M. genitalium colonization at the first prenatal visit was 8.4%. The incidence of low birth weight was 16.7%. The incidence of preterm delivery among women who were known to have a live birth was 16.7%. The incidence of preterm delivery did not differ with respect to M. genitalium colonization. The crude odds ratio for preterm delivery among women with M. genitalium colonization versus those without was 1.27 (95% confidence interval, 0.02–14.78). Conclusion M. genitalium colonization was not associated with preterm delivery among women with a high incidence of low birth weight newborns and preterm delivery, and a high prevalence of sexually transmitted infections. PMID:23834822

  12. Development and validation of a SIgA-ELISA for the detection of Mycoplasma hyopneumoniae infection.

    PubMed

    Feng, Zhi-Xin; Shao, Guo-Qing; Liu, Mao-Jun; Wang, Hai-Yan; Gan, Yuan; Wu, Xu-Su

    2010-07-14

    An alternative indirect enzyme-linked immunosorbent assay (ELISA) for the detection of Mycoplasma hyopneumoniae secretory IgA (SIgA) antibody (SIgA-ELISA) was developed using an adhesin (P97R1) of M. hyopneumoniae produced in Escherichia coli. The SIgA-ELISA assay was validated by the comparison with a nested-PCR assay and a commercial M. hyopneumoniae antibody detection kit (IgG-ELISA). Two hundred and sixty nasal swab samples, bronchoalveolar lavage fluids or serum samples were prepared for SIgA-ELISA validation from a M. hyopneumoniae-free farm, a M. hyopneumoniae vaccinated farm and two M. hyopneumoniae contaminated farms. The results showed that the SIgA-ELISA assay could distinguish the M. hyopneumoniae infection from M. hyopneumoniae vaccinated pigs, which was impossible for the current commercial M. hyopneumoniae antibody detection kits. The diagnostic sensitivity (DSN), specificity (DSP) and accuracy of the SIgA-ELISA were 97.0%, 94.4% and 95.8%, respectively and were compared with nested-PCR on 260 field nasal swab samples. The results of repeatability tests revealed that the coefficients of variation of swab samples within and between runs were less than 10%. This SIgA-ELISA is a needle-free detection methodology for large-scale surveys of M. hyopneumoniae infection. PMID:20053508

  13. Identification of Mycoplasma mycoides subsp. mycoides Small Colony Genes Coding for T-Cell Antigens ?

    PubMed Central

    Totté, Philippe; Mather, Arshad; Reslan, Lina; Boublik, Yvan; Niang, Mamadou; Du Plessis, Dion; Dedieu, Laurence

    2010-01-01

    Genes of the Mycoplasma mycoides subsp. mycoides small colony biotype (MmmSC) coding for proteins capable of eliciting protective T-cell memory responses have potential for incorporation into a recombinant subunit vaccine against contagious bovine pleuropneumonia (CBPP). Here we used lymphocytes from cattle that had completely recovered from infection to screen products of MmmSC genes for recognition by CD4+ effector memory (Tem) and central memory (Tcm) T lymphocytes. Six MmmSC genes (abc, gapN, glpO, lppA, lppB, and ptsG) were expressed as histidine-tagged recombinant polypeptides, or synthetic overlapping peptides, before inclusion in proliferation and gamma interferon (IFN-?) assays. Only two MmmSC antigens, LppA and PtsG, consistently induced recall proliferation from immune CD4+ T cells and IFN-? production in all animals tested. Moreover, LppA and PtsG were shown to possess epitopes recognized by both short-lived CD4+ Tem and long-lived CD4+ Tcm cells. PMID:20534794

  14. Mhp107 Is a Member of the Multifunctional Adhesin Family of Mycoplasma hyopneumoniae*

    PubMed Central

    Seymour, Lisa M.; Falconer, Linda; Deutscher, Ania T.; Minion, F. Chris; Padula, Matthew P.; Dixon, Nicholas E.; Djordjevic, Steven P.; Walker, Mark J.

    2011-01-01

    Mycoplasma hyopneumoniae is the causative pathogen of porcine enzootic pneumonia, an economically significant disease that disrupts the mucociliary escalator in the swine respiratory tract. Expression of Mhp107, a P97 paralog encoded by the gene mhp107, was confirmed using ESI-MS/MS. To investigate the function of Mhp107, three recombinant proteins, F1Mhp107, F2Mhp107, and F3Mhp107, spanning the N-terminal, central, and C-terminal regions of Mhp107 were constructed. Colonization of swine by M. hyopneumoniae requires adherence of the bacterium to ciliated cells of the respiratory tract. Recent studies have identified a number of M. hyopneumoniae adhesins that bind heparin, fibronectin, and plasminogen. F1Mhp107 was found to bind porcine heparin (KD ?90 nm) in a dose-dependent and saturable manner, whereas F3Mhp107 bound fibronectin (KD ?180 nm) at physiologically relevant concentrations. F1Mhp107 also bound porcine plasminogen (KD = 24 nm) in a dose-dependent and physiologically relevant manner. Microspheres coated with F3Mhp107 mediate adherence to porcine kidney epithelial-like (PK15) cells, and all three recombinant proteins (F1Mhp107-F3Mhp107) bound swine respiratory cilia. Together, these findings indicate that Mhp107 is a member of the multifunctional M. hyopneumoniae adhesin family of surface proteins and contributes to both adherence to the host and pathogenesis. PMID:21245147

  15. Mycoplasma genitalium infection among HIV-positive women: prevalence, risk factors and association with vaginal shedding.

    PubMed

    Gatski, M; Martin, D H; Theall, K; Amedee, A; Clark, R A; Dumestre, J; Chhabra, P; Schmidt, N; Kissinger, P

    2011-03-01

    This study examined the prevalence and factors associated with Mycoplasma genitalium (MG) infection among HIV-positive women and the association between MG and vaginal HIV-1 RNA shedding. HIV-positive women attending an outpatient clinic in New Orleans, Louisiana, USA, from 2002 to 2005 were examined for a battery of sexually transmitted infections (STIs) and underwent a behavioural survey. A selected subset had a measurement of vaginal shedding analysed. Of the 324 HIV-positive women, 32 (9.9%) were infected with MG. HIV-positive women with MG were more likely to be co-infected with Neisseria gonorrhoeae and Chlamydia trachomatis and to have had ?1 male sexual partners in the last month. In the subset (n = 164), no differences were found in the presence of detectable vaginal HIV-1 RNA between women infected and not infected with MG (30.8% versus 34.8% shedding; P = 0.69). While MG was a common co-STI in this sample of HIV-positive women, it was not associated with vaginal HIV shedding. PMID:21464453

  16. Seroprevalence of Mycoplasma pneumoniae in HIV-infected patients using a microparticle agglutination test.

    PubMed

    Shankar, Esaki Muthu; Kumarasamy, Nagalingeswaran; Balakrishnan, Pachamuthu; Vengatesan, A; Kownhar, Hayath; Solomon, Suniti; Rao, Usha Anand

    2006-06-01

    Mycoplasma pneumoniae is increasingly recognized as a common and important pathogen in community settings, and is responsible for various pulmonary and extrapulmonary conditions in the normal population. However, the seroepidemiology of acute M. pneumoniae infection in HIV-infected individuals is still unclear worldwide. This study examined the seroprevalence of antibodies to M. pneumoniae in HIV-infected patients admitted with respiratory complaints at a tertiary AIDS care centre in Chennai, India. A commercial gelatin microparticle agglutination test (Serodia-Myco II, Fujirebio) was used for the determination of antibodies against M. pneumoniae in acute serum specimens. Of the 200 HIV-infected patients with underlying pulmonary conditions tested, 34 (17 % positivity; 95 % CI 12-23 %) had antibodies specific to M. pneumoniae, while among the 40 patients with no underlying pulmonary symptoms, five (12.5 % positivity; 95 % CI 4-27 %) had evidence of anti-M. pneumoniae antibody. This shows that the incidence of M. pneumoniae seropositivity is greater in patients with underlying pulmonary complaints. Most positive titres were found in the age group 28-37 years in the symptomatic and symptom-free groups (64.7 and 60 %, respectively). The positive titres ranged from 40 to >20 480. High titres (> or =320) were found in 10 out of the 39 patients (25.6 %). This seroprevalence study reports a 16.2 % prevalence of M. pneumoniae infections in HIV-infected patients by a particle agglutination test. PMID:16687596

  17. Efficacy of early Mycoplasma hyopneumoniae vaccination against mixed respiratory disease in older fattening pigs.

    PubMed

    Del Pozo Sacristán, R; Sierens, A; Marchioro, S B; Vangroenweghe, F; Jourquin, J; Labarque, G; Haesebrouck, F; Maes, D

    2014-02-22

    The present field study investigated the efficacy of early Mycoplasma hyopneumoniae vaccination in a farrow-to-finish pig herd with respiratory disease late in the fattening period due to combined infections with M hyopneumoniae and viral pathogens. Five hundred and forty piglets were randomly divided into three groups of 180 piglets each: two groups were vaccinated (Stellamune Once) at either 7 (V1) or 21 days of age (V2), and a third group was left non-vaccinated (NV). The three treatment groups were housed in different pens within the same compartment during the nursery period, and were housed in different but identical compartments during the fattening period. The efficacy was evaluated using performance and pneumonia lesions. The average daily weight gain during the fattening period was 19 (V1) and 18 g/day (V2) higher in both vaccinated groups when compared with the NV group. However, the difference was not statistically significant (P>0.05). The prevalence of pneumonia was significantly lower in both vaccinated groups (V1: 71.5 and V2: 67.1 per cent) when compared with the NV group (80.2 per cent) (P<0.05). There were no significant differences between the two vaccination groups. In conclusion, in the present herd with respiratory disease during the second half of the fattening period caused by M hyopneumoniae and viral infections, prevalence of pneumonia lesions were significantly reduced and growth losses numerically (not statistically significant) decreased by both vaccination schedules. PMID:24436349

  18. [The frequency of detection of Mycoplasma meleagridis in breeding turkeys depending on the laying age].

    PubMed

    Rott, M; Pfützner, H; Gigas, H; Mach, B

    1989-01-01

    Coating of air sacs was recorded from day-old chicks from parents with Mycoplasma (M.) meleagridis infection, with positive findings being obtained from 9.52% of all animals early in the laying period and from 34.09% up to the 8th laying week. M. meleagridis was isolated from palatine and cloacal swabs taken of laying hens and insemination cocks, with positive findings being 50-60% prior to the laying period (28th week of age), 100% at start of laying, and 80% in the 14th laying week. M. meleagridis was identified in 50% of all embryonated eggs as of the 1st laying week and in 100% as of the 4th week. M. meleagridis was cultured from 30.67% of all sperm samples tested, between the 30th and 46th week of production. Differences were found to exist between individual cocks, with 4 cocks being without M. meleagridis at all. There was usually agreement between positive M. meleagridis findings from sperm and cloacal swabs. M. meleagridis was eliminated from cock sperm by spectinomycin (0.6 mg/ml diluting medium), but M. iowae was not. PMID:2619470

  19. Invasion of bovine peripheral blood mononuclear cells and erythrocytes by Mycoplasma bovis.

    PubMed

    van der Merwe, Jacques; Prysliak, Tracy; Perez-Casal, Jose

    2010-11-01

    Mycoplasma bovis is a small, cell wall-less bacterium that contributes to a number of chronic inflammatory diseases in both dairy and feedlot cattle, including mastitis and bronchopneumonia. Numerous reports have implicated M. bovis in the activation of the immune system, while at the same time inhibiting immune cell proliferation. However, it is unknown whether the specific immune-cell population M. bovis is capable of attaching to and potentially invading. Here, we demonstrate that incubation of M. bovis Mb1 with bovine peripheral blood mononuclear cells (PBMC) resulted in a significant reduction in their proliferative responses while still remaining viable and capable of gamma interferon secretion. Furthermore, we show that M. bovis Mb1 can be found intracellularly (suggesting a role for either phagocytosis or attachment/invasion) in a number of select bovine PBMC populations (T cells, B cells, monocytes, ?? T cells, dendritic cells, NK cells, cytotoxic T cells, and T-helper cells), as well as red blood cells, albeit it at a significantly lower proportion. M. bovis Mb1 appeared to display three main patterns of intracellular staining: diffuse staining, an association with the intracellular side of the cell membrane, and punctate/vacuole-like staining. The invasion of circulating immune cells and erythrocytes could play an important role in disease pathogenesis by aiding the transport of M. bovis from the lungs to other sites. PMID:20713619

  20. Climate Variability and Nonstationary Dynamics of Mycoplasma pneumoniae Pneumonia in Japan

    PubMed Central

    Onozuka, Daisuke; Chaves, Luis Fernando

    2014-01-01

    Background A stationary association between climate factors and epidemics of Mycoplasma pneumoniae (M. pneumoniae) pneumonia has been widely assumed. However, it is unclear whether elements of the local climate that are relevant to M. pneumoniae pneumonia transmission have stationary signatures of climate factors on their dynamics over different time scales. Methods We performed a cross-wavelet coherency analysis to assess the patterns of association between monthly M. pneumoniae cases in Fukuoka, Japan, from 2000 to 2012 and indices for the Indian Ocean Dipole (IOD) and El Niño Southern Oscillation (ENSO). Results Monthly M. pneumoniae cases were strongly associated with the dynamics of both the IOD and ENSO for the 1–2-year periodic mode in 2005–2007 and 2010–2011. This association was non-stationary and appeared to have a major influence on the synchrony of M. pneumoniae epidemics. Conclusions Our results call for the consideration of non-stationary, possibly non-linear, patterns of association between M. pneumoniae cases and climatic factors in early warning systems. PMID:24740102

  1. Detection of Mycoplasma genitalium from male primary urine specimens: an epidemiologic dichotomy with Trichomonas vaginalis.

    PubMed

    Napierala, Maureen; Munson, Erik; Wenten, David; Phipps, Paula; Gremminger, Roger; Schuknecht, Mary Kay; Munson, Kimber L; Boyd, Vivian; Hamer, Deb; Schell, Ronald F; Hryciuk, Jeanne E

    2015-07-01

    A total of 2750 male urines subjected to a transcription-mediated amplification (TMA)-based Mycoplasma genitalium assay yielded 188 positive results (6.84%). This rate was similar to Chlamydia trachomatis (6.87%; P=0.96) and greater than Neisseria gonorrhoeae (4.0%) and Trichomonas vaginalis (2.3%; P<0.0002). Mean age of M. genitalium-infected males (30.8) was similar to N. gonorrhoeae (P=0.78) but less than T. vaginalis (mean, 41.6; P<0.0001). A total of 266 STI clinic encounters had at least 1 sexually transmitted infection (STI); 36.5% of these encounters had sole detection of M. genitalium (P?0.009 versus sole detection of other STI agents). In 209 community encounters with at least 1 STI, 22.0% exhibited sole detection of M. genitalium (P=0.0007 versus sole M. genitalium detection in STI clinic males), while 18.7% had sole detection of T. vaginalis (P<0.0002 versus detection in STI clinic males). TMA-based M. genitalium screening identifies additional cases of nongonococcal urethritis. PMID:25934156

  2. Response of Airborne Mycoplasma pneumoniae to Abrupt Changes in Relative Humidity

    PubMed Central

    Hatch, M. T.; Wright, D. N.; Bailey, G. D.

    1970-01-01

    The effect of an abrupt change in the relative humidity on the viability of airborne Mycoplasma pneumoniae has been examined. When the microbial aerosols were permitted to equilibrate in air held at either low or high humidities and were then subjected to a sudden shift to a mid-range humidity, a significant loss (>90%) of the colony-forming units per liter of aerosol occurred within 8 min. In contrast, a change in the relative humidity of more than 18% in either direction from a lethal mid-range humidity noticeably decreased the rate of biological decay. Double humidity shifts (i.e., from dry to a mid-range level and then to a high humidity range) were very detrimental, with very few survivors after 8 min. These results indicate that the biological stability of airborne M. pneumoniae may be easily modified by a sudden change in the relative humidity, such as occurs in natural atmospheres. This increased sensitivity brought about by producing changes in relative humidity through the lethal humidity range may provide a method whereby the control of these organisms in naturally contaminated indoor air environments may be eventually achieved. PMID:5437301

  3. Effects of Mycoplasma pneumoniae infection on airway neurokinin-1 receptor expression in BALB/c mice.

    PubMed

    Zhang, H; Wei, B; Shang, Y X; Jiao, X Y; Wang, L; He, M B; Han, X H; Wang, G Z

    2014-01-01

    The aim of this study was to establish a BALB/c mouse model of Mycoplasma pneumoniae (MP) infection and to explore the expression of neurokinin-1 receptor (NK1-R) in the trachea and lung tissue and changes in its relative content at different time points (on the 3rd, 7th, 14th, 21st, and 30th days after infection) in MP-infected BALB/c mice. Immunohistochemistry and Western blot analysis were performed to determine NK1-R expression in the trachea and lung tissue and changes in relative content in MP-infected BALB/c mice. After MP infection, the expression of NK1-R on the surfaces of upper tracheal and bronchial epithelial cells, submucosa, and alveolar epithelial cells, as well as around the smooth muscle, was upregulated more significantly in the infection group than in the control group (P < 0.05); NK1-R protein expression was enhanced on the 3rd, 7th, 14th, 21st, and 30th days after infection compared with that of the control group (P < 0.05). NK1-R expression in the trachea, bronchus, and lung tissue increased in MP-infected BALB/c mice, which may explain why wheezing occurs after MP infection. PMID:25366726

  4. Mutagenesis, biochemical, and biophysical characterization of Mycoplasma arthritidis-derived mitogen

    PubMed Central

    Li, Hongmin; Zhao, Yiwei; Guo, Yi; VanVranken, Sandra J.; Li, Zhong; Eisele, Leslie; Mourad, Walid

    2014-01-01

    Mycoplasma arthritidis -derived mitogen (MAM) is a superantigen (SAg) that can activate large fractions of T cells bearing particular TCR V? elements. Here we report the mutagenesis, biochemical and biophysical studies on the dimerization of MAM in solution. Our studies showed that although MAM mainly exists as a monomer in solution, a small percentage of MAM molecules form homodimer at high protein concentration, regardless of the presence of Zn2+. A distinct peak corresponding to a MAM homodimer was detected in the presence of EDTA, using both chemical cross-linking and analytical ultracentrifugation methods. Further mutagenesis studies revealed that single mutation of residues at the interface of the crystallographic dimer of MAM does not significantly affect the dimerization of MAM in solution. Circular dichroism (CD) analysis indicated that addition of Zn2+ does not induce conformational changes of MAM from its apo-state. Thermal denaturation experiments indicated that addition of Zn2+ to MAM solution resulted in a decrease of melting point (Tm), whereas addition of EDTA did not affect the Tm of MAM. These results imply that there is no defined Zn2+-binding site on MAM. PMID:16753217

  5. Delineation of immunodominant and cytadherence segment(s) of Mycoplasma pneumoniae P1 gene

    PubMed Central

    2014-01-01

    Background Adhesion of Mycoplasma pneumoniae (M. pneumoniae) to host epithelial cells requires several adhesin proteins like P1, P30 and P116. Among these proteins, P1 protein has been inedited as one of the major adhesin and immunogenic protein present on the attachment organelle of M. pneumoniae. In the present study, we scanned the entire sequence of M. pneumoniae P1 protein to identify the immunodominant and cytadherence region(s). M. pneumoniae P1 gene was synthesized in four segments replacing all the UGA codons to UGG codons. Each of the four purified P1 protein fragment was analyzed for its immunogenicity with anti-M. pneumoniae M129 antibodies (Pab M129) and sera of M. pneumoniae infected patients by western blotting and ELISA. Antibodies were produced against all the P1 protein fragments and these antibodies were used for M. pneumoniae adhesion, M. pneumoniae adhesion inhibition and M. pneumoniae surface exposure assays using HEp-2 cells lines. Results Our results show that the immunodominant regions are distributed throughout the entire length of P1 protein, while only the N- and C- terminal region(s) of P1 protein are surface exposed and block cytadhesion to HEp-2 cells, while antibodies to two middle fragments failed to block cytadhesion. Conclusions These results have important implications in designing strategies to block the attachment of M. pneumoniae to epithelial cells, thus preventing the development of atypical pneumonia. PMID:24774062

  6. Bronchiolitis Associated With Mycoplasma Pneumoniae in Infants in Suzhou China Between 2010 and 2012

    PubMed Central

    Wang, Yuqing; Hao, Chuangli; Ji, Wei; Yan, Yongdong; Shao, Xuejun; Xu, Jun

    2015-01-01

    Viruses cause most cases of bronchiolitis in infants; consequently the importance of other agents such as Mycoplasma pneumoniae (MP) in the etiology of bronchiolitis may not be fully recognized. We investigated the prevalence and seasonal distribution of bronchiolitis caused by MP in 674 children admitted to the Children's Hospital affiliated with Soochow University from January 2010 to December 2012. The presence of MP was confirmed by real-time PCR. During the 3 years, we identified MP in 17.2% of the children with bronchiolitis. The annual MP detection rates were 16.6% in 2010, 17.8% in 2011, and 17.2% in 2012. MP was detected throughout the year, with a peak from July to September. The median age of MP-positive children was 10 months. Common clinical manifestations included cough, wheezing, and high fever. Moist and/or wheezing rales were frequent, and pulmonary interstitial infiltration was seen in 66.4% of chest X-rays. Patients with MP infection were older, were more likely to have pulmonary interstitial infiltration, and had shorter hospital stays than those with respiratory syncytial virus infection. Our study revealed MP as an important cause of bronchiolitis, with peaks of occurrence during the summer and early autumn. Pulmonary interstitial infiltrations were a common event. PMID:25597274

  7. Hamiltonians of strain effects

    NASA Astrophysics Data System (ADS)

    Suzuki, Tatsuo

    2001-12-01

    Hamiltonians that generally describe the effects of strain are proposed. The strain effects can be calculated easily from the unstrained potential using these Hamiltonians. These Hamiltonians are valid when the strain is spatially modulated, and are also valid when the strain exists in a magnetic field. These Hamiltonians can also be used in the improved effective mass approximation.

  8. Short term exposure to NO sup 2 decreases intrapulmonary killing of Mycoplasma pulmonis by damaging alveolar macrophages

    SciTech Connect

    Davis, J.K.; Davidson, M.K.; Schoeb, T.R.; Lindsey, J.R. (Univ. of Alabama, Birmingham (United States) Veteran Administration Medical Center, Birmingham, AL (United States))

    1991-03-11

    Previous studies have shown that exposure of pathogen free C57BL/6N mice to 5 or 10 ppm of NO{sub 2} increased severity of murine respiratory mycoplasmosis and that this effect was associated with decreased intrapulmonary killing (IPK) of Mycoplasma pulmonis (MP). The purposes of the present studies were to titrate the NO{sub 2} effect and to determine if the changes in IPK were due to the effects of NO{sub 2} on alveolar macrophages. Exposure to less than 5 ppm NO{sub 2} had no effect on IPK of MP. Bronchoalveolar lavage (BAL) cells killed MP in vitro only if they were allowed to associate with mycoplasmas in vivo. Prior exposure to NO{sub 2} abrogated killing in this in vivo-in vitro model. Exposure to NO{sub 2} did not increase the protein content of BAL within 24 hours. Greater than 95% of the BAL cells were macrophages, and greater than 98% of the cell-associated mycoplasmas were on or in alveolar macrophages. Immediately after exposure, viability of alveolar macrophages, as measured by trypan blue exclusion and fluorescein diacetate uptake, was 89 {plus minus} 4% and 88 {plus minus} 4% in the control group, respectively; 56 {plus minus} 19% and 64 {plus minus} 11% in the group receiving MP alone; 23 {plus minus} 7% and 48 {plus minus} 9% in the group receiving 10 ppm NO{sub 2}; and 16 {plus minus} 6% and 25 {plus minus} 6% in the group receiving both MP and NO{sub 2} exposures. Viability was significantly decreased following exposure to 5 or 10 ppm NO{sub 2}, but not following exposure to 2 ppm. Viability did not return to normal until 7 days after exposure to NO{sub 2}, at which time IPK also returned to normal. The cellular target of NO{sub 2} exposure in relation to IPK of MP appears to be the alveolar macrophage.

  9. 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 glands with increased mucous secretion was obvious. Air sacs showed thickening with presence of inflammatory cells and edema. PMID:25910901

  10. Identification of a 521-kilodalton protein (Gli521) involved in force generation or force transmission for Mycoplasma mobile gliding.

    PubMed

    Seto, Shintaro; Uenoyama, Atsuko; Miyata, Makoto

    2005-05-01

    Several mycoplasma species are known to glide on solid surfaces such as glass in the direction of the membrane protrusion, but the mechanism underlying this movement is unknown. To identify a novel protein involved in gliding, we raised monoclonal antibodies against a detergent-insoluble protein fraction of Mycoplasma mobile, the fastest glider, and screened the antibodies for inhibitory effects on gliding. Five monoclonal antibodies stopped the movement of gliding mycoplasmas, keeping them on the glass surface, and all of them recognized a large protein in immunoblotting. This protein, named Gli521, is composed of 4,738 amino acids, has a predicted molecular mass of 520,559 Da, and is coded downstream of a gene for another gliding protein, Gli349, which is known to be responsible for glass binding during gliding. Edman degradation analysis indicated that the N-terminal region is processed at the peptide bond between the amino acid residues at positions 43 and 44. Analysis of gliding mutants isolated previously revealed that the Gli521 protein is missing in a nonbinding mutant, m9, where the gli521 gene is truncated by a nonsense mutation at the codon for the amino acid at position 1170. Immunofluorescence and immunoelectron microscopy indicated that Gli521 localizes all around the base of the membrane protrusion, at the "neck," as previously observed for Gli349. Analysis of the inhibitory effects of the anti-Gli521 antibody on gliding motility revealed that this protein is responsible for force generation or force transmission, a role distinct from that of Gli349, and also suggested conformational changes of Gli349 and Gli521 during gliding. PMID:15866938

  11. Mycoplasma ovipneumoniae--a primary cause of severe pneumonia epizootics in the Norwegian Muskox (Ovibos moschatus) population.

    PubMed

    Handeland, Kjell; Tengs, Torstein; Kokotovic, Branko; Vikøren, Turid; Ayling, Roger D; Bergsjø, Bjarne; Sigurðardóttir, Olöf G; Bretten, Tord

    2014-01-01

    The Norwegian muskox (Ovibos moschatus) population lives on the high mountain plateau of Dovre and originates from animals introduced from Greenland. In the late summers of 2006 and 2012, severe outbreaks of pneumonia with mortality rates of 25-30% occurred. During the 2012 epidemic high quality samples from culled sick animals were obtained for microbiological and pathological examinations. High throughput sequencing (pyrosequencing) of pneumonic lung tissue revealed high concentrations of Mycoplasma ovipneumoniae in all six animals examined by this method and Pasteurella multocida subsp. multocida in four animals, whereas no virus sequences could be identified. Mycoplasma ovipneumoniae and P. multocida multocida were also isolated by culture. Using real time PCR on lung swabs, M. ovipneumoniae was detected in all of the 19 pneumonic lungs examined. Gross pathological examination revealed heavy consolidations primarily in the cranial parts of the lungs and it also identified one case of otitis media. Histologically, lung lesions were characterized as acute to subacute mixed exudative and moderately proliferative bronchoalveolar pneumonia. Immunohistochemical (IHC) examination revealed high load of M. ovipneumoniae antigens within lung lesions, with particularly intensive staining in the neutrophils. Similar IHC finding were observed in archived lung tissue blocks from animals examined during the 2006 epidemic. An M. ovipneumoniae specific ELISA was applied on bio-banked muskox sera from stray muskoxen killed in the period 2004-2013 and sick muskoxen culled, as well as sera from wild reindeer (Rangifer tarandus tarandus) on Dovre and muskoxen from Greenland. Serology and mycoplasma culturing was also carried out on sheep that had been on pasture in the muskox area during the outbreak in 2012. Our findings indicated separate introductions of M. ovipneumoniae infection in 2006 and 2012 from infected co-grazing sheep. Salt licks shared by the two species were a possible route of transmitting infection. PMID:25198695

  12. In vitro and in vivo cell invasion and systemic spreading of Mycoplasma agalactiae in the sheep infection model.

    PubMed

    Hegde, Shivanand; Hegde, Shrilakshmi; Spergser, Joachim; Brunthaler, René; Rosengarten, Renate; Chopra-Dewasthaly, Rohini

    2014-11-01

    Generally regarded as extracellular pathogens, molecular mechanisms of mycoplasma persistence, chronicity and disease spread are largely unknown. Mycoplasma agalactiae, an economically important pathogen of small ruminants, causes chronic infections that are difficult to eradicate. Animals continue to shed the agent for several months and even years after the initial infection, in spite of long antibiotic treatment. However, little is known about the strategies that M. agalactiae employs to survive and spread within an immunocompetent host to cause chronic disease. Here, we demonstrate for the first time its ability to invade cultured human (HeLa) and ruminant (BEND and BLF) host cells. Presence of intracellular mycoplasmas is clearly substantiated using differential immunofluorescence technique and quantitative gentamicin invasion assays. Internalized M. agalactiae could survive and exit the cells in a viable state to repopulate the extracellular environment after complete removal of extracellular bacteria with gentamicin. Furthermore, an experimental sheep intramammary infection was carried out to evaluate its systemic spread to organs and host niches distant from the site of initial infection. Positive results obtained via PCR, culture and immunohistochemistry, especially the latter depicting the presence of M. agalactiae in the cytoplasm of mammary duct epithelium and macrophages, clearly provide the first formal proof of M. agalactiae's capability to translocate across the mammary epithelium and systemically disseminate to distant inner organs. Altogether, the findings of these in vitro and in vivo studies indicate that M. agalactiae is capable of entering host cells and this might be the strategy that it employs at a population level to ward off the host immune response and antibiotic action, and to disseminate to new and safer niches to later egress and once again proliferate upon the return of favorable conditions to cause persistent chronic infections. PMID:25129554

  13. Housing Conditions Modulate the Severity of Mycoplasma pulmonis Infection in Mice Deficient in Class A Scavenger Receptor

    PubMed Central

    Booth, Jennifer L; Umstead, Todd M; Hu, Sanmei; Dybvig, Kevin F; Cooper, Timothy K; Wilson, Ronald P; Chroneos, Zissis C

    2014-01-01

    Mycoplasmosis is a frequent causative microbial agent of community-acquired pneumonia and has been linked to exacerbation of chronic obstructive pulmonary disease. The macrophage class A scavenger receptor (SRA) facilitates the clearance of noxious particles, oxidants, and infectious organisms by alveolar macrophages. We examined wildtype and SRA?/? mice, housed in either individually ventilated or static filter-top cages that were cycled with fresh bedding every 14 d, as a model of gene–environment interaction on the outcome of pulmonary Mycoplasma pulmonis infection. Intracage NH3 gas measurements were recorded daily prior to infection. Mice were intranasally infected with 1 × 107 cfu M. pulmonis UAB CT and evaluated at 3, 7, and 14 d after inoculation. Wildtype mice cleared 99.5% of pulmonary M. pulmonis by 3 d after infection but remained chronically infected through the study. SRA?/? mice were chronically infected with 40-fold higher mycoplasma numbers than were wildtype mice. M. pulmonis caused a chronic mixed inflammatory response that was accompanied with high levels of IL1?, KC, MCP1, and TNF? in SRA?/? mice, whereas pulmonary inflammation in WT mice was represented by a monocytosis with elevation of IL1?. Housing had a prominent influence on the severity and persistence of mycoplasmosis in SRA?/? mice. SRA-/- mice housed in static cages had an improved recovery and significant changes in surfactant proteins SPA and SPD compared with baseline levels. These results indicate that SRA is required to prevent chronic mycoplasma infection of the lung. Furthermore, environmental conditions may exacerbate chronic inflammation in M. pulmonis-infected SRA?/? mice. PMID:25527023

  14. Superlattice strain gage

    DOEpatents

    Noel, Bruce W. (Espanola, NM); Smith, Darryl L. (Los Alamos, NM); Sinha, Dipen N. (Los Alamos, NM)

    1990-01-01

    A strain gage comprising a strained-layer superlattice crystal exhibiting piezoelectric properties is described. A substrate upon which such a strained-layer superlattice crystal has been deposited is attached to an element to be monitored for strain. A light source is focused on the superlattice crystal and the light reflected from, passed through, or emitted from the crystal is gathered and compared with previously obtained optical property data to determine the strain in the element.

  15. Superlattice strain gage

    DOEpatents

    Noel, B.W.; Smith, D.L.; Sinha, D.N.

    1988-06-28

    A strain gage comprising a strained-layer superlattice crystal exhibiting piezoelectric properties is described. A substrate upon which such a strained-layer superlattice crystal has been deposited is attached to an element to be monitored for strain. A light source is focused on the superlattice crystal and the light reflected from, passed through, or emitted from the crystal is gathered and compared with previously obtained optical property data to determine the strain in the element. 8 figs.

  16. Continuous Regional Arterial Infusion Therapy for Acute Necrotizing Pancreatitis Due to Mycoplasma pneumoniae Infection in a Child

    SciTech Connect

    Nakagawa, Motoo, E-mail: lmloltlolol@gmail.com; Ogino, Hiroyuki; Shimohira, Masashi; Hara, Masaki; Shibamoto, Yuta [Nagoya City University Graduate School of Medical Sciences, Department of Radiology (Japan)

    2009-05-15

    A case of acute necrotizing pancreatitis due to Mycoplasma pneumoniae infection was treated in an 8-year-old girl. She experienced acute pancreatitis during treatment for M. pneumoniae. Contrast-enhanced computed tomographic scan revealed necrotizing pancreatitis. The computed tomographic severity index was 8 points (grade E). A protease inhibitor, ulinastatin, was provided via intravenous infusion but was ineffective. Continuous regional arterial infusion therapy was provided with gabexate mesilate (FOY-007, a protease inhibitor) and meropenem trihydrate, and the pancreatitis improved. This case suggests that infusion therapy is safe and useful in treating necrotizing pancreatitis in children.

  17. Results of molecular detection of Mycoplasma pneumoniae among patients with acute respiratory infection and in their household contacts reveals children as human reservoirs

    Microsoft Academic Search

    J. W. Dorigo-Zetsma; Berry Wilbrink; Jacob Dankert

    2001-01-01

    During a 30-month prospective study in The Netherlands, the distribution of Mycoplasma pneumoniae and respiratory viruses among 1172 patients with acute respiratory infection (ARI) who were treated in the outpatient general practitioner setting was studied. M. pneumoniae, as detected by polymerase chain reaction analysis, was present in 39 (3.3%) patients. The infection rate was similar in all age groups. Nose

  18. Porcine circovirus type 2 (PCV2) vaccination is effective in reducing disease and PCV2 shedding in semen of boars concurrently infected with PCV2 and Mycoplasma hyopneumoniae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objectives of this study were to determine if the amount of porcine circovirus type 2 (PCV2) shed in semen will be increased in boars experimentally coinfected with Mycoplasma hyopneumoniae (MHYO) and if PCV2 vaccination of the boars prior to PCV2 exposure will result in reduced PCV2 viremia and...

  19. First isolation of Mycoplasma capricolum subsp. capricolum, one of the causal agents of caprine contagious agalactia, on the island of Lanzarote (Spain)

    Microsoft Academic Search

    C. De la Fe; A. Gutiérrez; J. B. Poveda; P. Assunção; A. S. Ramírez; F. Fabelo

    2007-01-01

    During an unusually long period of bad weather, several outbreaks of caprine contagious agalactia (CCA) were reported in a number of flocks on the island of Lanzarote (Canary Islands, Spain). Clinical and subclinical mastitis in lactating goats and some cases of arthritis and pneumonia in kids were observed in the affected flocks. Mycoplasma capricolum subsp. capricolum was isolated as the

  20. Gamma interferon-producing CD4 T-cells correlate with resistance to Mycoplasma mycoides subsp. mycoides S.C. infection in cattle

    Microsoft Academic Search

    L. Dedieu; V. Balcer-Rodrigues; A. Yaya; B. Hamadou; O. Cisse; M. Diallo; M. Niang

    2005-01-01

    Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides SC (MmmSC), is one of the most significant cattle disease in Africa. The control measures, which led to eradication from numerous countries are not feasible in Africa where the only prophylaxis relies on vaccination. However, the attenuated vaccines, used up to now in Africa, are of low efficiency. The development