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Sample records for mycoplasma hominis strains

  1. Detection and prevention of mycoplasma hominis infection

    DOEpatents

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

    1997-01-21

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

  2. Identification and characterization of novel Mycoplasma spp. belonging to the hominis group from griffon vultures.

    PubMed

    Lecis, R; Chessa, B; Cacciotto, C; Addis, M F; Coradduzza, E; Berlinguer, F; Muzzeddu, M; Lierz, M; Carcangiu, L; Pittau, M; Alberti, A

    2010-08-01

    Mycoplasmas are commensals and pathogens of various avian species, and are also regularly found in birds of prey, although their significance to birds' health remains unclear. Here we describe two novel Mycoplasma isolated from the upper respiratory tract of four Eurasian griffon vultures (Gyps fulvus) housed in a wildlife recovery centre in Sardinia (Italy). By sequencing the 16S rRNA gene and the entire 16S/23S intergenic spacer region, the new strains were classified within the Mycoplasma taxonomy at the group and cluster levels, showing that the two isolates fall into the Mycoplasma synoviae and Mycoplasma hominis clusters of the hominis group, respectively. We combined molecular tools and immunoblotting methods in order to further characterize these isolates, and antigenic analyses overall confirmed the molecular findings. Different levels of pathogenicity and prevalence of these strains might have different implications for the conservation and reintroduction of vultures.

  3. Entry and intracellular location of Mycoplasma hominis in Trichomonas vaginalis.

    PubMed

    Vancini, Ricardo Gomes; Benchimol, Marlene

    2008-01-01

    The parasite Trichomonas vaginalis causes one of the most common non-viral sexually transmitted infections in humans. The coexistence of different sexually transmitted diseases in the same individual is very common, such as vaginal infections by T. vaginalis in association with Mycoplasma fermentans or Mycoplasma hominis. However, the consequences and behavior of mycoplasma during trichomonad infections are virtually unknown. This study was undertaken to elucidate whether mycoplasmas enter and leave trichomonad cells and if so how. M. hominis was analyzed in different trichomonad isolates and the process of internalization and the pathway within the parasite was studied. Parasites naturally and experimentally infected with mycoplasmas were used and transmission electron microscopy, cytochemistry and PCR analyses were performed. The results show that: (1) M. hominis enters T. vaginalis cells by endocytosis; (2) some mycoplasmas use a terminal polar tip as anchor to the trichomonad plasma membrane; (3) some trichomonad isolates are able to digest mycoplasmas, mainly when the trichomonads are experimentally infected; (4) some fresh virulent isolates are able to maintain mycoplasmas as cohabitants in the cell's interior; (5) some mycoplasmas are able to escape from the vacuole to the trichomonad cytosol, and trichomonad plasma membrane budding suggested that mycoplasmas could leave the parasite cell. PMID:17710384

  4. Possible involvement of Mycoplasma hominis in inhibiting the formation of biofilms by uropathogenic Escherichia coli (UPEC).

    PubMed

    Oh, Sangnam; Go, Gwang-Woong; Choi, Nag-Jin; Oh, Sejong; Kim, Younghoon

    2013-01-01

    Here we examined the involvement of Mycoplasma hominis in the formation of biofilms by uropathogenic Escherichia coli (UPEC) strain CFT073. Initially, we thought that M. hominis does not affect the fitness of UPEC, including the growth and production of signaling molecules, such as autoinducer-2 and indole. We found, however, that the presence of M. hominis significantly decreased the degree of biofilm formation by UPEC CFT073 (approximately a 60% reduction for 10(5) ccu/mL of M. hominis as compared with UPEC alone). We also found that it had a slight effect in inhibiting the attachment and cytotoxicity of UPEC CFT073. These findings are specific to these UPEC strains rather than to enterohemorrhagic E. coli (EHEC) strains, found in normal intestinal flora. In addition, we performed whole-transcriptome profiling and quantitative real-time polymerase chain reaction (qRT-PCR) analysis. This indicated that the PhoPQ system and the anti-termination protein (encoded by ybcQ) were involved in the reduction of biofilm formation by M. hominis (corroborated by qRT-PCR). Furthermore, our results indicate that M. hominis raises the degree of transcription of toxin genes, including hha and pasT. Hence, we suggest a possible role of M. hominis in affecting the formation of biofilms by UPEC in the urinary tract.

  5. RESPIRATORY PATHWAYS IN THE MYCOPLASMA. II. PATHWAY OF ELECTRON TRANSPORT DURING OXIDATION OF REDUCED NICOTINAMIDE ADENINE DINUCLEOTIDE BY MYCOPLASMA HOMINIS.

    PubMed

    VANDEMARK, P J; SMITH, P F

    1964-07-01

    VanDemark, P. J. (University of South Dakota, Vermillion), and P. F. Smith. Respiratory pathways in the Mycoplasma. II. Pathway of electron transport during oxidation of reduced nicotinamide adenine dinucleotide by Mycoplasma hominis. J. Bacteriol. 88:122-129. 1964.-Unlike the flavin-terminated respiratory pathway of the fermentative Mycoplasma, the respiratory chain of the nonfermentative M. hominis strain 07 appears to be more complex, involving quinones and cytochromes in addition to flavins. In addition to reduction by reduced nicotine adenine dinucleotide (NADH) and reduced nicotine adenine dinucleotide phosphate, nonpyridine nucleotide-linked reduction of the respiratory chain of this organism occurred with succinate, lactate, and short-chained acyl coenzyme A derivatives as electron donors. Enzymes catalyzing the oxidation of NADH included an NADH oxidase, a diaphorase, a quinone reductase, and a cytochrome c reductase. The oxidation of NADH was sensitive to a variety of inhibitors, including 10(-4)m Atabrine, 10(-3)m sodium amytal, 10(-5)mp-chloromercuribenzoate, 10(-4)m antimycin A, and 10(-4)m potassium cyanide. The oxidase was resolved by the addition of 5% trichloroacetic acid and reactivated by the addition of flavin adenine dinucleotide but not flavin mononucleotide. The M. hominis sonic extract contained an NADH-coenzyme Q reductase. The oxidation of NADH was stimulated by the addition of either menadione or vitamin K(2) (C(35)). The oxidase was inactivated by extraction with ether or irradiation at 360 mmu. The ether-inactivated enzyme was partially reactivated by the addition of "lipid" extract of the enzyme and coenzyme Q(6). Difference spectra of the cell extracts revealed the presence of "b" and "a" type cytochromes. These cell extracts were found to contain a cyanide-and azide-sensitive cytochrome oxidase and catalase. PMID:14197876

  6. Prevalence and Antibiotic Susceptibility of Mycoplasma hominis and Ureaplasma urealyticum in Pregnant Women

    PubMed Central

    Lee, Min Young; Lee, Woo In; Kang, So Young; Jeon, You La

    2016-01-01

    Mycoplasma hominis (M. hominis) and Ureaplasma urealyticum (U. urealyticum) are important opportunistic pathogens that cause urogenital infections and complicate pregnancy. The aim of this study was to investigate the prevalence, effects on pregnancy outcomes, and antimicrobial susceptibilities of M. hominis and U. urealyticum. We tested vaginal swabs obtained from 1035 pregnant women for the presence of genital mycoplasmas between June 2009 and May 2014. The laboratory and clinical aspects of genital mycoplasmas infection were reviewed retrospectively, and the identification and antimicrobial susceptibility of genital mycoplasmas were determined using the Mycoplasma IST-2 kit. A total of 571 instances of M. hominis and/or U. urealyticum were detected. Of them, M. hominis was detected in two specimens, whereas U. urealyticum was detected in 472 specimens. The remaining 97 specimens were positive for both M. hominis and U. urealyticum. Preterm deliveries were frequently observed in cases of mixed infection of M. hominis and U. urealyticum, and instances of preterm premature rupture of membrane were often found in cases of U. urealyticum. The rates of non-susceptible isolates to erythromycin, empirical agents for pregnant women, showed increasing trends. In conclusion, the prevalence of M. hominis and/or U. urealyticum infections in pregnant women is high, and the resistance rate of antimicrobial agents tends to increase. Therefore, to maintain a safe pregnancy, it is important to identify the isolates and use appropriate empirical antibiotics immediately. PMID:27401661

  7. [Effects of the symbiosis of Trichomonas vaginalis with Mycoplasma hominis on ferredoxin gene].

    PubMed

    Liu, Xiaodong; Wen, Wenjing; Xue, Changgui

    2011-08-01

    We isolated 30 Trichomonas vaginalis for the PCR detection from the gynecological outpatients in the Affiliated Hospital of Zhengzhou University using the specific 16s rDNA primers of Mycoplasma hominis. The results showed that there were 25 cases of Mycoplasma hominis infection, with the infection rate of 83.33%. This gave a clew that the symbiosis of Trichomonas vaginalis with Mycoplasma hominis may be of certain generality in China. We sequenced the ferredoxin gene of 10 Trichomonas vaginalis where 5 Mycoplasma hominis were positive and five negative, and found that the ferredoxin (Fd) gene of the 10 Trichomonas vaginalis were exactly the same. But compared to the genes in the GenBank, a comparative analysis of the gene revealed that there were 3 more ctg bases at the 200th position of encoding leucine, but this did not lead to changes in reading frame. The gene homology was 99%.

  8. Epidemiology of Ureaplasma urealyticum and Mycoplasma hominis in the semen of male outpatients with reproductive disorders

    PubMed Central

    Zhu, Xiaofei; Li, Min; Cao, Huiling; Yang, Xuewen; Zhang, Chunbing

    2016-01-01

    The aim of the present study was to investigate the association between Mycoplasma infection and infertility in male outpatients among a Chinese population. Epidemiological data, including prevalence, age distribution and antibiotic resistance profile of patients with an Ureaplasma urealyticum or Mycoplasma hominis infection were collected between 2009 and 2012. Among the 7,374 individuals analyzed, 3,225 patients (43.7%) were determined to be positive for infection with U. urealyticum, M. hominis or for both Mycoplasmas. Among the positive cultures, U. urealyticum was detected most frequently, while M. hominis was rarely found. The age range of 25–34 years was the preferred period for the positive detection. Tetracyclines and josamycin were the most effective agents against both genital Mycoplasmas, including in the case of co-infection. Macrolides (erythromycin, roxithromycin, azithromycin, clarithromycin except for josamycin) were effective against the majority of U. urealyticum clinical isolates, but were naturally resisted by M. hominis in this study. Fluoroquinolones had the lowest activity against U. urealyticum, particularly in cases of M. hominis co-infection. Furthermore, fluoroquinolones showed a similar pattern of drug resistance against M. hominis to that of U. urealyticum. Antibiotic resistance did not vary significantly over the test period. Notably, an elevated multi-drug resistance rate was observed in patients co-infected with both Mycoplasmas. In light of the epidemiological characteristics of genital Mycoplasmas in male infertility patients, the present results may aid Chinese clinicians to implement rational drug usage and avoid the overuse of antibiotics. PMID:27443698

  9. Mycoplasma hominis and Trichomonas vaginalis: a unique case of symbiotic relationship between two obligate human parasites.

    PubMed

    Dessì, Daniele; Rappelli, Paola; Diaz, Nicia; Cappuccinelli, Piero; Fiori, Pier Luigi

    2006-01-01

    Mollicutes are the smallest and simplest self-replicating microorganisms. Despite the minimal genome and apparent lack of complexity, mycoplasmas show a high degree of adaptation to the most diverse environments. Mycoplasma hominis is a human sexually transmitted mycoplasma which is able to establish a biological association with Trichomonas vaginalis, a pathogenic flagellated protist. M. hominis and T. vaginalis share the same specific natural niche, the human genitourinary tract. Symbiotic relationships between unicellular eukaryotes and bacteria are well known and have been extensively studied, providing interesting insights into the biology of one or both the symbionts. The relationship between T. vaginalis and M. hominis is unique in that it was the first described association of two obligated human parasites. Several aspects of this relationship have been investigated, showing how the trichomonad may be viewed not only as a new niche for M. hominis, but also as a "Trojan horse" for the transmission of the bacterial infection to the human host. PMID:16720288

  10. Standardized methods and quality control limits for agar and broth microdilution susceptibility testing of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum.

    PubMed

    Waites, Ken B; Duffy, Lynn B; Bébéar, Cécile M; Matlow, Anne; Talkington, Deborah F; Kenny, George E; Totten, Patricia A; Bade, Donald J; Zheng, Xiaotian; Davidson, Maureen K; Shortridge, Virginia D; Watts, Jeffrey L; Brown, Steven D

    2012-11-01

    An international multilaboratory collaborative study was conducted to develop standard media and consensus methods for the performance and quality control of antimicrobial susceptibility testing of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum using broth microdilution and agar dilution techniques. A reference strain from the American Type Culture Collection was designated for each species, which was to be used for quality control purposes. Repeat testing of replicate samples of each reference strain by participating laboratories utilizing both methods and different lots of media enabled a 3- to 4-dilution MIC range to be established for drugs in several different classes, including tetracyclines, macrolides, ketolides, lincosamides, and fluoroquinolones. This represents the first multilaboratory collaboration to standardize susceptibility testing methods and to designate quality control parameters to ensure accurate and reliable assay results for mycoplasmas and ureaplasmas that infect humans.

  11. Survey on association between Mycoplasma hominis endocervical infection and spontaneous abortion using Polymerase Chain Reaction

    PubMed Central

    Farhadifar, Fariba; Khodabandehloo, Mazaher; Ramazanzadeh, Rashid; Rouhi, Samaneh; Ahmadi, Amjad; Ghaderi, Ebrahim; Roshani, Daem; Soofizadeh, Nasrin; Rezzaii, Masoomeh

    2016-01-01

    Background: Mycoplasma infections are suggested as etiology of adverse pregnancy outcomes. Objective: The aim of this study was to evaluate the association of Mycoplasma hominis (M. hominis) infection and spontaneous abortion among pregnant women. Materials and Methods: In this case-control study that was conducted from August 2012 to January 2013, totally, 109 women were included with spontaneous abortion with gestational ages of 10-20 weeks (Cases), and 109 women with normal pregnancy with gestational ages between 20-37 weeks (Controls) in Sanandaj, Iran. Using specific primers and extracted DNA from endocervical swabs, a PCR test was conducted for detection of M. hominis infection in women. For comparison of qualitative and quantitative variables, independent Fisher tests were used and p<0.05 was considered significant. Results: The total frequency of M. hominis infection was 6 (2.75%) in women. The frequency of M. hominis infection was 2 (1.83%) in the case group (spontaneous abortion) and 4 (3.66%) in the control group, respectively. In both case and control groups, no association was seen between M.hominis infection and spontaneous abortion (OR=0. 49, CI 95%: 0.08-2.73, p=0. 683). Conclusion: M. hominis was positive in the genital tract of some pregnant women, but it was not associated with spontaneous abortion. However, to prevent adverse pregnancy outcomes in women, foetus and neonate, routine screening and treatment for the genital Mycoplasma is recommended. PMID:27294216

  12. An epidemiological survey of Mycoplasma hominis and Ureaplasma urealyticum in gynaecological outpatients, Rome, Italy.

    PubMed

    Verteramo, R; Patella, A; Calzolari, E; Recine, N; Marcone, V; Osborn, J; Chiarini, F; Degener, A M

    2013-12-01

    The objective of this study was to assess the prevalence of Ureaplasma urealyticum and Mycoplasma hominis infections and to investigate associations between their presence in the lower female genital tract and lifestyle characteristics. The study was performed on a population of 3115 women, comparing the demographic and behavioural characteristics of 872 women with U. urealyticum infection and 142 women with M. hominis with uninfected women, using univariate and multiple logistic regression analysis. The prevalence of infection with U. urealyticum was 28% and M. hominis was 4.6%. In multivariate logistic regression analysis, intrauterine device, number of sexual partners and age (<35 years) were significantly associated with U. urealyticum while previous induced abortion, condom use and young age at first intercourse (<16 years) were associated with M. hominis infection. U. urealyticum infection presents the same demographic and behavioural characteristics of a sexually transmitted disease. The unprotective role of condom use suggests a non-sexual mode of transmission of M. hominis infection.

  13. Phylogeny of some mycoplasmas from ruminants based on 16S rRNA sequences and definition of a new cluster within the hominis group.

    PubMed

    Pettersson, B; Uhlén, M; Johansson, K E

    1996-10-01

    Almost complete (> 96%) 16S rRNA sequences from nine ruminant mycoplasmas have been determined by solid-phase DNA sequencing. Polymorphisms were found in four of the 16S rRNA sequences, which indicated the existence of two different rRNA operons. Seven polymorphisms were found in Mycoplasma agalatiae, three were found in Mycoplasma bovis, one was found in Mycoplasma alkalescens, and one was found in Mycoplasma bovirhinis. The sequence data were used for construction of phylogenetic trees. All but one of the ruminant mycoplasmas sequenced in this work clustered in the hominis group. A close relationship was found between M. agalactiae and M. bovis, with a 99% nucleotide similarity between their 16S rRNA sequences. They were also found to be members of the Mycoplasma lipophilum cluster of the hominis group. Furthermore, the 16S rRNA comparisons showed that Mycoplasma alkalescens and Mycoplasma canadense are closely related (> 98.5%), and these species were found to cluster in the Mycoplasma hominis cluster of the hominis group. Interestingly, M. bovirhinis grouped in a new phylogenetic cluster of the hominis group. The new cluster, which was supported by bootstrap percentage values, signature nucleotide analysis, and higher-order structural elements, was named the Mycoplasma synoviae cluster. Mycoplasma bovoculi, Mycoplasma conjunctivae, and Mycoplasma ovipneumoniae clustered in the Mycoplasma neurolyticum cluster of the hominis group. Mycoplasma alvi clustered with Mycoplasma pirum in the M. pneumoniae cluster of the pneumoniae group.

  14. The presence of Mycoplasma hominis in isolates of Trichomonas vaginalis impacts significantly on DNA fingerprinting results.

    PubMed

    Xiao, J C; Xie, L F; Zhao, L; Fang, S L; Lun, Z R

    2008-03-01

    The genetic characterization of Trichomonas vaginalis (Protista: Trichomonadidae), the causative agent of trichomoniasis in humans, is central to understanding the epidemiology, treatment, drug resistance, and virulence as well as the diagnosis and control of this parasite. Various molecular approaches, including DNA fingerprinting, have been employed for this purpose, and random amplification of polymorphic DNA (RAPD) continues to be utilized. However, little attention has been paid to the fact that some T. vaginalis populations can harbor symbiotic Mycoplasma hominis and/or other agents, which could cause artifacts in the RAPD results. In the present study, we demonstrate clearly that the presence of M. hominis from T. vaginalis isolates impacts significantly on RAPD results and on the subsequent analyses and interpretation of data sets. Moreover, symbiotic M. hominis displays an isolate-to-isolate variability in RAPD profile before elimination, suggesting a variability of M. hominis infection. PMID:18058131

  15. Prevalence of Ureaplasma urealyticum and Mycoplasma hominis in High Vaginal Swab Samples of Infertile Females

    PubMed Central

    Seifoleslami, Mehri; Safari, Aghdas; Khayyat Khameneie, Maryam

    2015-01-01

    Background: Mycoplasma hominis and Ureaplasma urealyticum are important causative agents of vaginitis, cervicitis, postpartum sepsis, reproductive infections and infertility in both males and females. Objectives: According to the uncertain prevalence of U. urealyticum and M. hominis in Iranian infertile females, the present study was carried out to determine the prevalence of U. urealyticum and M. hominis in high vaginal swab samples of fertile and infertile females. Patients and Methods: A total of 350 high vaginal swab specimens were taken from fertile and infertile females. Samples were cultured and those that were positive for bacteria were subjected to the polymerase chain reaction (PCR) for further confirmation. Results: Of the 350 collected samples, eleven were positive for M. hominis (3.14%), fifteen were positive for U. urealyticum (4.28%) and five were positive for both of them (1.42%). Prevalence of U. urealyticum and M. hominis in the high vaginal parts of infertile females was higher than fertile females (P < 0.05). The results of traditional method were also confirmed, using the PCR amplification of urease gene of U. urealyticum and 16SrRNA gene of the M. hominis. Ureaplasma urealyticum and M. hominis had a higher prevalence in the high vaginal samples collected during the summer season. Conclusions: Considerable prevalence of M. hominis and U. urealyticum in the high vaginal swab samples of infertile females compared to the low prevalence in fertile females may suggest that these two pathogens can be cause infertility. Application of the PCR method is recommended for rapid and sensitive detection of M. hominis and U. urealyticum in high vaginal swab samples. PMID:26756000

  16. Mycoplasma hominis in Cuban Trichomonas vaginalis isolates: association with parasite genetic polymorphism.

    PubMed

    Fraga, Jorge; Rodríguez, Nadia; Fernández, Carmen; Mondeja, Brian; Sariego, Idalia; Fernández-Calienes, Aymé; Rojas, Lazara

    2012-07-01

    Trichomonas vaginalis can be naturally infected with intracellular Mycoplasma hominis. This bacterial infection may have implications for trichomonal virulence and disease pathogenesis. The objective of the study was to report the presence of M. hominis in Cuban T. vaginalis isolates and to describe the association between the phenotype M. hominis infected with RAPD genetic polymorphism of T. vaginalis. The Random Amplified Polymorphic DNA (RAPD) technique was used to determine genetic differences among 40 isolates of T. vaginalis using a panel of 30 random primers and these genetic data were correlated with the infection of isolates with M. hominis. The trees drawn based on RAPD data showed no relations with metronidazole susceptibility and significantly association with the presence of M. hominis (P=0.043), which demonstrates the existence of concordance between the genetic relatedness and the presence of M. hominis in T. vaginalis isolates. This result could point to a predisposition of T. vaginalis for the bacterial enters and/or survival. PMID:22584035

  17. Selection of Mycoplasma hominis PG21 deletion mutants by cultivation in the presence of monoclonal antibody 552.

    PubMed Central

    Jensen, L T; Ladefoged, S; Birkelund, S; Christiansen, G

    1995-01-01

    Three mutants of Mycoplasma hominis PG21 were isolated and shown to contain alterations in the size of a repeat-containing gene encoding a surface-localized 135-kDa antigen designated Lmp1. The mutants were isolated by cultivating M. hominis for a 3-month period in the presence of Lmp1-specific monoclonal antibody (MAb) 552. The epitope for MAb 552 was localized at the repeated part of the protein. The gene encoding Lmp1 is part of a transcriptional complex that contains 9.5 direct repeats of 471 bp each. Pure cultures of mutant strains were obtained by subcloning, and three mutants were characterized. The mutants showed deletions of a various number of repeats. The deletions were accompanied by a decrease in size of the proteins. With increasing size of deletions, agglutination and growth inhibition by MAb 552 became less pronounced. Spontaneous aggregation of the mutant M. hominis cells in culture medium was, however, increased, indicating that the repeated elements may be of importance for repulsion of the cells. PMID:7543881

  18. 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

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

    PubMed Central

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

    2015-01-01

    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

  20. Symbiotic Association with Mycoplasma hominis Can Influence Growth Rate, ATP Production, Cytolysis and Inflammatory Response of Trichomonas vaginalis

    PubMed Central

    Margarita, Valentina; Rappelli, Paola; Dessì, Daniele; Pintus, Gianfranco; Hirt, Robert P.; Fiori, Pier L.

    2016-01-01

    The symbiosis between the parasitic protist Trichomonas vaginalis and the opportunistic bacterium Mycoplasma hominis is the only one currently described involving two obligate human mucosal symbionts with pathogenic capabilities that can cause independent diseases in the same anatomical site: the lower urogenital tract. Although several aspects of this intriguing microbial partnership have been investigated, many questions on the influence of this symbiosis on the parasite pathobiology still remain unanswered. Here, we examined with in vitro cultures how M. hominis could influence the pathobiology of T. vaginalis by investigating the influence of M. hominis on parasite replication rate, haemolytic activity and ATP production. By comparing isogenic mycoplasma-free T. vaginalis and parasites stably associated with M. hominis we could demonstrate that the latter show a higher replication rate, increased haemolytic activity and are able to produce larger amounts of ATP. In addition, we demonstrated in a T. vaginalis-macrophage co-culture system that M. hominis could modulate an aspect of the innate immuno-response to T. vaginalis infections by influencing the production of nitric oxide (NO) by human macrophages, with the parasite-bacteria symbiosis outcompeting the human cells for the key substrate arginine. These results support a model in which the symbiosis between T. vaginalis and M. hominis influences host-microbes interactions to the benefit of both microbial partners during infections and to the detriment of their host. PMID:27379081

  1. Symbiotic Association with Mycoplasma hominis Can Influence Growth Rate, ATP Production, Cytolysis and Inflammatory Response of Trichomonas vaginalis.

    PubMed

    Margarita, Valentina; Rappelli, Paola; Dessì, Daniele; Pintus, Gianfranco; Hirt, Robert P; Fiori, Pier L

    2016-01-01

    The symbiosis between the parasitic protist Trichomonas vaginalis and the opportunistic bacterium Mycoplasma hominis is the only one currently described involving two obligate human mucosal symbionts with pathogenic capabilities that can cause independent diseases in the same anatomical site: the lower urogenital tract. Although several aspects of this intriguing microbial partnership have been investigated, many questions on the influence of this symbiosis on the parasite pathobiology still remain unanswered. Here, we examined with in vitro cultures how M. hominis could influence the pathobiology of T. vaginalis by investigating the influence of M. hominis on parasite replication rate, haemolytic activity and ATP production. By comparing isogenic mycoplasma-free T. vaginalis and parasites stably associated with M. hominis we could demonstrate that the latter show a higher replication rate, increased haemolytic activity and are able to produce larger amounts of ATP. In addition, we demonstrated in a T. vaginalis-macrophage co-culture system that M. hominis could modulate an aspect of the innate immuno-response to T. vaginalis infections by influencing the production of nitric oxide (NO) by human macrophages, with the parasite-bacteria symbiosis outcompeting the human cells for the key substrate arginine. These results support a model in which the symbiosis between T. vaginalis and M. hominis influences host-microbes interactions to the benefit of both microbial partners during infections and to the detriment of their host. PMID:27379081

  2. The resistance analysis of Ureaplasma urealyticum and Mycoplasma hominis in female reproductive tract specimens.

    PubMed

    Ye, Guangyong; Jiang, Zhou; Wang, Min; Huang, Jiamin; Jin, Guochen; Lu, Shiming

    2014-01-01

    The aim of this study was to analyze the drug resistance of Ureaplasma urealyticum (Uu) and Mycoplasma hominis (Mh) in female reproductive track from 2007 to 2011 in Hangzhou. Antibiotics sensitivity test in Mycoplasma, which was isolated in clinics from 2007 to 2011 were analyzed retrospectively. The detection of Mycoplasma during 2007-2011 was 20,146 (54.37 %), of which the single infection rate of Uu was 42.08 %, of Mh 1.26 %, and of Uu+Mh was 11.02 %. The drug resistance rate of Uu was increased significantly in ofloxacin in 2007 (41.80 %), 2008 (45.94 %), 2009 (46.07 %), 2010 (50.36 %), and 2011 (53.22 %) (P < 0.05). The resistance rate to ciprofloxacin was significantly increased in 2007 (67.15 %), 2008 (67.44 %), 2009 (73.00 %), 2010 (75.28 %), and 2011 (75.28 %) (P < 0.05). Exceptionally, the resistance rates of the other antibiotics were low. The drug resistance rate of Uu was significantly increased with quinolones at increasing tendency. It is necessary to monitor the local drug resistance rate of Uu regularly to provide reasonable guidelines in clinics.

  3. Mycoplasma hominis septic arthritis in a pediatric renal transplant recipient: case report and review of the literature.

    PubMed

    Mian, Ayesa N; Farney, Alan C; Mendley, Susan R

    2005-01-01

    Septic arthritis (SA) typically occurs in young children, often from Staphylococcus. With chronic immunosuppression, however, pathogens may be atypical. A 15-year-old African-American female developed Mycoplasma hominis SA in her right hip 2 months following cadaveric renal transplant (Tx). Her presentation was subtle and indolent, without fever or leukocytosis. Although reported in adult Tx recipients, M. hominis infections have not been described in pediatric recipients. Early immunosuppression (basiliximab, prednisone, tacrolimus, mycophenolate mofetil and Thymoglobulin) may have increased her susceptibility to M. hominis. Optimal therapy for M. hominis SA is not well established and relapses occur. This patient underwent joint incision and drainage, treatment for 8 weeks with doxycycline and levofloxacin guided by in vitro sensitivities, and a reduction in immunosuppression. She has been free of ongoing infection for 3 years with stable graft function (Cr 1.1 mg/dL) on moderate immunosuppression with prednisone, tacrolimus and MMF.

  4. Comparative analysis of male and female populations on prevalence and antibiotic resistance of Mycoplasma hominis in China, 2005-2014.

    PubMed

    Kong, Yingying; Qiao, Yingli; Song, Jingjuan; Ruan, Zhi; Fei, Chunrong; Huang, Jun; Song, Tiejun; Jin, Hong; Ding, Honghui; Xie, Xinyou; Zhang, Jun

    2016-09-01

    The aim of this study was to estimate the prevalence and antimicrobial resistance rate of Mycoplasma hominis among male and female populations. A total of 67921 individuals were examined. All samples were isolated from patients at an outpatient clinic from January 2005 to December 2014. Species identification and antibiotic susceptibility testing were performed using Mycoplasma IST2. In this study, 523 (0.8%) and 4625 (6.8%) cultures, respectively, were positive for single M. hominis identification and simultaneous identification of both M. hominis and Ureaplasma spp. The results showed that both single and simultaneous identification were more frequent in the female than the male population. Moreover, the highest positive rates of single M. hominis identification were observed in 56-60-year-old males and 61-65-year-old females, and the rates of simultaneous identification were high in males aged >65 years and females aged 15-20 years. Among the seven antibiotics assessed, tetracycline, josamycin, doxycycline and pristinamycin were the most effective, whilst clarithromycin, ciprofloxacin and ofloxacin displayed extremely high resistance rates. Different antimicrobial susceptibility rates were observed between the two sexes, and the resistance rates to ofloxacin showed a significant difference (P<0.05). In conclusion, this study demonstrates that the prevalence of M. hominis varied significantly between the two sexes in the past 10 years and that the optimal antimicrobial therapy strategy should be directed by local susceptibility testing. PMID:27530842

  5. Computational prediction of Mycoplasma hominis proteins targeting in nucleus of host cell and their implication in prostate cancer etiology.

    PubMed

    Khan, Shahanavaj; Zakariah, Mohammed; Palaniappan, Sellappan

    2016-08-01

    Cancer has long been assumed to be a genetic disease. However, recent evidence supports the enigmatic connection of bacterial infection with the growth and development of various types of cancers. The cause and mechanism of the growth and development of prostate cancer due to Mycoplasma hominis remain unclear. Prostate cancer cells are infected and colonized by enteroinvasive M. hominis, which controls several factors that can affect prostate cancer growth in susceptible persons. We investigated M. hominis proteins targeting the nucleus of host cells and their implications in prostate cancer etiology. Many vital processes are controlled in the nucleus, where the proteins targeting M. hominis may have various potential implications. A total of 29/563 M. hominis proteins were predicted to target the nucleus of host cells. These include numerous proteins with the capability to alter normal growth activities. In conclusion, our results emphasize that various proteins of M. hominis targeted the nucleus of host cells and were involved in prostate cancer etiology through different mechanisms and strategies.

  6. Clonal analysis of Staphylococcus hominis strains isolated from hospitalized patients.

    PubMed

    Szczuka, Ewa; Trawczyński, Krzysztof; Kaznowski, Adam

    2014-01-01

    Staphylococcus hominis is a part of normal skin flora, but it is also a cause of nosocomial infections. The aim of this study was to investigate the genetic relatedness of 62 strains of S. hominis obtained from hospitalised patients during an 11-year period. For the discrimination of these clinical strains we used repetitive sequence-based PCR method (BOX-PCR) and multiple-locus variable-number tandem repeat analysis (MLVA). BOX-PCR analysis revealed a large genetic diversity among clinical strains and we did not find a predominant clone with the ability to persist in a hospital environment. MLVA is not as discriminatory as BOX fingerprinting and would not be a useful method for epidemiological studies.

  7. High rates of double-stranded RNA viruses and Mycoplasma hominis in Trichomonas vaginalis clinical isolates in South Brazil.

    PubMed

    da Luz Becker, Débora; dos Santos, Odelta; Frasson, Amanda Piccoli; de Vargas Rigo, Graziela; Macedo, Alexandre José; Tasca, Tiana

    2015-08-01

    Trichomonas vaginalis is the etiological agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in world, with 276.4 million new cases each year. T. vaginalis can be naturally infected with Mycoplasma hominis and Trichomonasvirus species. This study aimed to evaluate the prevalence of T. vaginalis infected with four distinct T. vaginalis viruses (TVVs) and M. hominis among isolates from patients in Porto Alegre city, South Brazil. An additional goal of this study was to investigate whether there is association between metronidazole resistance and the presence of M. hominis during TVV infection. The RNA expression level of the pyruvate ferredoxin oxidoreductase (PFOR) gene was also evaluated among metronidazole-resistant and metronidazole-sensitive T. vaginalis isolates. A total of 530 urine samples were evaluated, and 5.7% samples were positive for T. vaginalis infection. Among them, 4.51% were isolated from female patients and 1.12% were from male patients. Remarkably, the prevalence rates of M. hominis and TVV-positive T. vaginalis isolates were 56.7% and 90%, respectively. Most of the T. vaginalis isolates were metronidazole-sensitive (86.7%), and only four isolates (13.3%) were resistant. There is no statistically significant association between infection by M. hominis and infection by TVVs. Our results refute the hypothesis that the presence of the M. hominis and TVVs is enough to confer metronidazole resistance to T. vaginalis isolates. Additionally, the role of PFOR RNA expression levels in metronidazole resistance as the main mechanism of resistance to metronidazole could not be established. This study is the first report of the T. vaginalis infection by M. hominis and TVVs in a large collection of isolates from South Brazil.

  8. High rates of double-stranded RNA viruses and Mycoplasma hominis in Trichomonas vaginalis clinical isolates in South Brazil.

    PubMed

    da Luz Becker, Débora; dos Santos, Odelta; Frasson, Amanda Piccoli; de Vargas Rigo, Graziela; Macedo, Alexandre José; Tasca, Tiana

    2015-08-01

    Trichomonas vaginalis is the etiological agent of trichomoniasis, the most common non-viral sexually transmitted disease (STD) in world, with 276.4 million new cases each year. T. vaginalis can be naturally infected with Mycoplasma hominis and Trichomonasvirus species. This study aimed to evaluate the prevalence of T. vaginalis infected with four distinct T. vaginalis viruses (TVVs) and M. hominis among isolates from patients in Porto Alegre city, South Brazil. An additional goal of this study was to investigate whether there is association between metronidazole resistance and the presence of M. hominis during TVV infection. The RNA expression level of the pyruvate ferredoxin oxidoreductase (PFOR) gene was also evaluated among metronidazole-resistant and metronidazole-sensitive T. vaginalis isolates. A total of 530 urine samples were evaluated, and 5.7% samples were positive for T. vaginalis infection. Among them, 4.51% were isolated from female patients and 1.12% were from male patients. Remarkably, the prevalence rates of M. hominis and TVV-positive T. vaginalis isolates were 56.7% and 90%, respectively. Most of the T. vaginalis isolates were metronidazole-sensitive (86.7%), and only four isolates (13.3%) were resistant. There is no statistically significant association between infection by M. hominis and infection by TVVs. Our results refute the hypothesis that the presence of the M. hominis and TVVs is enough to confer metronidazole resistance to T. vaginalis isolates. Additionally, the role of PFOR RNA expression levels in metronidazole resistance as the main mechanism of resistance to metronidazole could not be established. This study is the first report of the T. vaginalis infection by M. hominis and TVVs in a large collection of isolates from South Brazil. PMID:26160539

  9. Dialysis Culture of T-Strain Mycoplasmas

    PubMed Central

    Masover, Gerald K.; Hayflick, Leonard

    1974-01-01

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

  10. Long-term survival and intracellular replication of Mycoplasma hominis in Trichomonas vaginalis cells: potential role of the protozoon in transmitting bacterial infection.

    PubMed

    Dessì, Daniele; Delogu, Giuseppe; Emonte, Eleonora; Catania, Maria Rosaria; Fiori, Pier Luigi; Rappelli, Paola

    2005-02-01

    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. By using gentamicin protection assays, double immunofluorescence, and confocal microscopy, we obtained strong evidence that M. hominis is located within protozoan cells. 5-Bromodeoxyuridine incorporation assays showed that intracellularly located mycoplasmas actively synthesize DNA. Our results demonstrate that M. hominis has the capability of entering trichomonad cells and of replicating inside the protozoon. These findings suggest that symbiosis might provide the bacteria, during human infection, with the capability to resist to environmental stresses, such as host defense mechanisms and pharmacological therapies. PMID:15664961

  11. Fatal nosocomial meningitis caused by Mycoplasma hominis in an adult patient: case report and review of the literature.

    PubMed

    Reissier, Sophie; Masson, Romain; Guérin, François; Viquesnel, Gérald; Petitjean-Lecherbonnier, Joëlle; Pereyre, Sabine; Cattoir, Vincent; Isnard, Christophe

    2016-07-01

    Meningitis due to Mycoplasma hominis in adults is rarely described, with only three cases having been reported to date. A case of fatal meningitis in a 39-year-old patient after a neurosurgical procedure for a subarachnoid haemorrhage is reported herein. Identification and treatment were significantly delayed because of the rarity of the aetiology and difficulty identifying this organism with the routinely used conventional methods, such as Gram staining and agar growth on standard agar plates. Clinical procedures and the treatment of 'culture-negative' central nervous system infections is a real challenge for clinical microbiologists and clinicians, and M. hominis has to be considered as a potential, although very uncommon, pathogen. PMID:27208637

  12. Antibiotic Susceptibility Profiles of Mycoplasma Hominis and Ureaplasma Urealyticum Isolated During a Population-Based Study Concerning Women Infertility in Northeast Romania

    PubMed Central

    Mihai, Mareş; Valentin, Năstasă; Bogdan, Doroftei; Carmen, Chifiriuc Mariana; Coralia, Bleotu; Demetra, Socolov

    2011-01-01

    The study was carried out on 1068 infertile women under initial evaluation. For Mycoplasma hominis, the highest resistance rates were registered for ciprofloxacin (72.22%), followed by macrolides and ofloxacin. For Ureaplasma urealyticum, the ciprofloxacin resistance was also high (51.72%), while the resistance rates to other tested antibiotics were significantly lower. PMID:24031629

  13. Genetic Passive Immunization with Adenoviral Vector Expressing Chimeric Nanobody-Fc Molecules as Therapy for Genital Infection Caused by Mycoplasma hominis

    PubMed Central

    Dolzhikova, Inna V.; Shcherbinin, Dmitry N.; Zubkova, Olga V.; Ivanova, Tatiana I.; Tukhvatulin, Amir I.; Shmarov, Maxim M.; Logunov, Denis Y.; Naroditsky, Boris S.; Gintsburg, Aleksandr L.

    2016-01-01

    Developing pathogen-specific recombinant antibody fragments (especially nanobodies) is a very promising strategy for the treatment of infectious disease. Nanobodies have great potential for gene therapy application due to their single-gene nature. Historically, Mycoplasma hominis has not been considered pathogenic bacteria due to the lack of acute infection and partially due to multiple studies demonstrating high frequency of isolation of M. hominis samples from asymptomatic patients. However, recent studies on the role of latent M. hominis infection in oncologic transformation, especially prostate cancer, and reports that M. hominis infects Trichomonas and confers antibiotic resistance to Trichomonas, have generated new interest in this field. In the present study we have generated specific nanobody against M. hominis (aMh), for which the identified target is the ABC-transporter substrate-binding protein. aMh exhibits specific antibacterial action against M. hominis. In an attempt to improve the therapeutic properties, we have developed the adenoviral vector-based gene therapy approach for passive immunization with nanobodies against M. hominis. For better penetration into the mucous layer of the genital tract, we fused aMh with the Fc-fragment of IgG. Application of this comprehensive approach with a single systemic administration of recombinant adenovirus expressing aMh-Fc demonstrated both prophylactic and therapeutic effects in a mouse model of genital M. hominis infection. PMID:26962869

  14. Genetic Passive Immunization with Adenoviral Vector Expressing Chimeric Nanobody-Fc Molecules as Therapy for Genital Infection Caused by Mycoplasma hominis.

    PubMed

    Burmistrova, Daria A; Tillib, Sergey V; Shcheblyakov, Dmitry V; Dolzhikova, Inna V; Shcherbinin, Dmitry N; Zubkova, Olga V; Ivanova, Tatiana I; Tukhvatulin, Amir I; Shmarov, Maxim M; Logunov, Denis Y; Naroditsky, Boris S; Gintsburg, Aleksandr L

    2016-01-01

    Developing pathogen-specific recombinant antibody fragments (especially nanobodies) is a very promising strategy for the treatment of infectious disease. Nanobodies have great potential for gene therapy application due to their single-gene nature. Historically, Mycoplasma hominis has not been considered pathogenic bacteria due to the lack of acute infection and partially due to multiple studies demonstrating high frequency of isolation of M. hominis samples from asymptomatic patients. However, recent studies on the role of latent M. hominis infection in oncologic transformation, especially prostate cancer, and reports that M. hominis infects Trichomonas and confers antibiotic resistance to Trichomonas, have generated new interest in this field. In the present study we have generated specific nanobody against M. hominis (aMh), for which the identified target is the ABC-transporter substrate-binding protein. aMh exhibits specific antibacterial action against M. hominis. In an attempt to improve the therapeutic properties, we have developed the adenoviral vector-based gene therapy approach for passive immunization with nanobodies against M. hominis. For better penetration into the mucous layer of the genital tract, we fused aMh with the Fc-fragment of IgG. Application of this comprehensive approach with a single systemic administration of recombinant adenovirus expressing aMh-Fc demonstrated both prophylactic and therapeutic effects in a mouse model of genital M. hominis infection. PMID:26962869

  15. Biofilm formation by Staphylococcus hominis strains isolated from human clinical specimens.

    PubMed

    Szczuka, Ewa; Telega, Kinga; Kaznowski, Adam

    2015-01-01

    Staphylococcus hominis is the third species of coagulase-negative staphylococci (CoNS) most frequently isolated from specimens of patients with hospital-acquired infections. Many infections caused by CoNS appeared to be associated with biofilms. Nevertheless, the knowledge of the ability of S. hominis to form a biofilm is limited. The aim of this study was to analyze the formation of the biofilm by 56 S. hominis strains isolated from clinical cases. The biofilm three-dimensional structure was reconstructed by confocal laser scanning microscopy. We found that most of S. hominis strains carried icaADBC genes encoding polysaccharide intercellular adhesin (PIA), which plays a crucial role in the formation of biofilms in staphylococci strains. However, only a half of the ica-positive strains had an ability to form a biofilm in vitro. In this study, we also accessed the sensitivity of biofilms of S. hominis strains to sodium metaperiodate, proteinase K and DNase. We found that polysaccharides and proteins are the major components of the extracellular matrix of the biofilm formed by S. hominis. DNase did not have a significant effect on biofilms, which suggested that nucleic acid plays a minor role in the mature biofilm.

  16. Swine and poultry pathogens: the complete genome sequences of two strains of Mycoplasma hyopneumoniae and a strain of Mycoplasma synoviae.

    PubMed

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

    2005-08-01

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

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

    PubMed Central

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

    2005-01-01

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

  18. Proteomic analysis of Mycoplasma gallisepticum vaccine strain F

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  19. Occurrence of Urease in T Strains of Mycoplasma

    PubMed Central

    Shepard, Maurice C.; Lunceford, Carl D.

    1967-01-01

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

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

    PubMed Central

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

    1993-01-01

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

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

    PubMed

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

    1993-02-01

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

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

    PubMed

    Cousin, Sylvie; Creno, Sophie; Ma, Laurence; Clermont, Dominique; Loux, Valentin; Bizet, Chantal; Bouchier, Christiane

    2013-01-01

    We report the draft genome sequence of the strain Lactobacillus hominis CRBIP 24.179(T), isolated from a human clinical sample. The total length of the 28 contigs is about 1.9 Mb, with a G+C content of 37% and 1,983 coding sequences.

  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. Genome Sequence of a Mycoplasma meleagridis Field Strain

    PubMed Central

    Bertolotti, Luigi; Catania, Salvatore; Pourquier, Philippe; Rosati, Sergio

    2016-01-01

    Mycoplasma meleagridis is a major cause of disease and economic loss in turkeys. Here, we report the genome sequence of an M. meleagridis field strain, which enlarges the knowledge about this bacterium and helps the identification of possible coding sequences for drug resistance genes and specific antigens. PMID:26941131

  5. Isolation of Mycoplasma genitalium strains from the male urethra.

    PubMed Central

    Jensen, J S; Hansen, H T; Lind, K

    1996-01-01

    Mycoplasma genitalium is a human mycoplasma species which, on the basis of detection by PCR, has been incriminated as a cause of nongonococcal urethritis. Previously, only two strains from the urogenital tract and five strains from extragenital sites have been isolated. We have developed a method for the isolation of this fastidious microbe. M. genitalium from PCR-positive urethral specimens was initially propagated in Vero cell cultures grown in serum-free medium supplemented with Ultroser HY serum substitute. Growth was monitored by PCR. The M. genitalium strains grown in cell cultures could subsequently be subcultured in modified Friis's FF broth medium. Several passages in broth medium were required before growth on agar medium was attained. A total of 11 urethral specimens positive for M. genitalium by PCR from male patients with urethritis were investigated. Six strains were adapted to growth in broth medium, and four of these strains were cloned. Three specimens were overgrown by other mycoplasmas during propagation in the cell cultures. In only two PCR-positive specimens was propagation of M. genitalium unsuccessful. The use of cell culture combined with PCR monitoring of mycoplasmal growth may prove to be more widely applicable for the isolation of other fastidious mollicutes. PMID:8789002

  6. 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

  7. DNA probes for the detection of mycoplasmas in genital specimens.

    PubMed

    Roberts, M C; Hooton, M; Stamm, W; Holmes, K K; Kenny, G E

    1987-06-01

    The utility of whole-genomic DNA probes for the detection of infections by genital mycoplasmas was investigated in 220 men attending a sexually transmitted diseases clinic. In 144 patients, probe results were compared with quantitative culture results. The prevalence of Mycoplasma hominis was 11% by culture, whereas the prevalence of ureaplasmas was 38%. The M. hominis DNA probe detected 9 of 16 M. hominis culture-positive specimens and 2 of 128 culture-negative specimens. The Ureaplasma urealyticum DNA probe detected 36 of 57 U. urealyticum culture-positive specimens and 18 of 87 culture-negative specimens. Most of the probe-negative culture-positive specimens had colony counts of less than 10(3) organisms/ml of specimen. The DNA probe does not require viable organisms, and the probe-positive, culture-negative specimens suggest that false-negative cultures occurred, perhaps due to specimen handling or insensitivity of culture methods for some strains of mycoplasmas.

  8. MLVA typing of Mycoplasma hyopneumoniae bacterins and field strains

    PubMed Central

    Tamiozzo, P.; Zamora, R.; Lucchesi, P. M. A.; Estanguet, A.; Parada, J.; Carranza, A.; Camacho, P.; Ambrogi, A.

    2015-01-01

    Because of the lack of information about both the genetic characteristics of Mycoplasma hyopneumoniae commercial vaccines and their relationship with field strains, the authors attempted to identify genetic subtypes of some M hyopneumoniae bacterins, and to compare them with M. hyopneumoniae field strains. Six commercial M hyopneumoniae bacterins and 28 bronchoalveolar lavages from pigs at slaughter from three herds were analysed by Multiple-Locus Variable number tandem repeat Analysis (MLVA) on p146R1, p146R3, H4, H5 and p95 loci. The results obtained showed the presence of more than one M hyopneumoniae genotype in some pigs and also in one of the bacterins analysed. It is also worth noting that MLVA typing allowed the distinction among circulating field strains and also when comparing them with vaccine strains, which, knowing the relatedness among them, could be useful in the research of the efficacy of the vaccines. PMID:26495127

  9. MLVA typing of Mycoplasma hyopneumoniae bacterins and field strains.

    PubMed

    Tamiozzo, P; Zamora, R; Lucchesi, P M A; Estanguet, A; Parada, J; Carranza, A; Camacho, P; Ambrogi, A

    2015-01-01

    Because of the lack of information about both the genetic characteristics of Mycoplasma hyopneumoniae commercial vaccines and their relationship with field strains, the authors attempted to identify genetic subtypes of some M hyopneumoniae bacterins, and to compare them with M. hyopneumoniae field strains. Six commercial M hyopneumoniae bacterins and 28 bronchoalveolar lavages from pigs at slaughter from three herds were analysed by Multiple-Locus Variable number tandem repeat Analysis (MLVA) on p146R1, p146R3, H4, H5 and p95 loci. The results obtained showed the presence of more than one M hyopneumoniae genotype in some pigs and also in one of the bacterins analysed. It is also worth noting that MLVA typing allowed the distinction among circulating field strains and also when comparing them with vaccine strains, which, knowing the relatedness among them, could be useful in the research of the efficacy of the vaccines. PMID:26495127

  10. Molecular basis of resistance to macrolides, lincosamides and streptogramins in Staphylococcus hominis strains isolated from clinical specimens.

    PubMed

    Szczuka, Ewa; Makowska, Nicoletta; Bosacka, Karolina; Słotwińska, Anna; Kaznowski, Adam

    2016-03-01

    Coagulase-negative staphylococci (CoNS) are the most frequently isolated bacteria from the blood and the predominant cause of nosocomial infections. Macrolides, lincosamides and streptogramin B (MLSB) antibiotics, especially erythromycin and clindamycin, are important therapeutic agents in the treatment of methicillin-resistant staphylococci infections. Among CoNS, Staphylococcus hominis represents the third most common organism. In spite of its clinical significance, very little is known about its mechanisms of resistance to antibiotics, especially MLSB. Fifty-five S. hominis isolates from the blood and the surgical wounds of hospitalized patients were studied. The erm(C) gene was predominant in erythromycin-resistant S. hominis isolates. The methylase genes, erm(A) and erm(B), were present in 15 and 25% of clinical isolates, respectively. A combination of various erythromycin resistance methylase (erm) genes was detected in 15% S. hominis isolates. The efflux gene msr(A) was detected in 18% of isolates, alone in four isolates, and in different combinations in a further six. The lnu(A) gene, responsible for enzymatic inactivation of lincosamides was carried by 31% of the isolates. No erythromycin resistance that could not be attributed to the genes erm(A), erm(B), erm(C) and msr(A) was detected. In S. hominis, 75 and 84%, respectively, were erythromycin resistant and clindamycin susceptible. Among erythromycin-resistant S. hominis isolates, 68% of these strains showed the inducible MLSB phenotype. Four isolates harbouring the msr(A) genes alone displayed the MSB phenotype. These studies indicated that resistance to MLSB in S. hominis is mostly based on the ribosomal target modification mechanism mediated by erm genes, mainly the erm(C), and enzymatic drug inactivation mediated by lnu(A).

  11. Chronic Endometritis and Positive Mycoplasma Cultures: Is There a Correlation?

    PubMed Central

    Nyirjesy, Paul; Amin-Hanjani, Soheil

    1995-01-01

    Objective: This study was undertaken to assess the impact of mycoplasma strains (Mycoplasma hominis or Ureaplasma urealyticum) on the development of chronic endometritis. Methods: Fifty-eight patients with acute pelvic infection were enrolled in this prospective cohort study. Endometrial cultures and biopsies were obtained on admission and 5–7 and 21–28 days after completion of treatment. Results: Of 148 samples, 40 were positive for mycoplasma strains (group A) and 58 were positive for mycoplasma with other pathogens (group B). Twenty-seven samples were positive for other pathogens only (group C). Chronic endometritis was seen in 7 (17.5%), 30 (51.7%), and 10 (37%) in group A, B, and C patients, respectively. Conclusions: The presence of mycoplasma strains in the endometrial cavity was not found to be associated with an increased incidence of chronic endometritis. PMID:18475413

  12. Draft Genome Sequence of a Cardiobacterium hominis Strain Isolated from Blood Cultures of a Patient with Infective Endocarditis

    PubMed Central

    Tagini, Florian; Pillonel, Trestan; Asner, Sandra; Prod’hom, Guy

    2016-01-01

    Cardiobacterium hominis is a well-known commensal bacterium of the oral cavity and an agent of infective endocarditis in humans. Here, we provide a draft genome sequence of a pathogenic strain isolated from blood cultures of a patient with infectious endocarditis. PMID:27660783

  13. Draft Genome Sequence of a Cardiobacterium hominis Strain Isolated from Blood Cultures of a Patient with Infective Endocarditis.

    PubMed

    Tagini, Florian; Pillonel, Trestan; Asner, Sandra; Prod'hom, Guy; Greub, Gilbert

    2016-01-01

    Cardiobacterium hominis is a well-known commensal bacterium of the oral cavity and an agent of infective endocarditis in humans. Here, we provide a draft genome sequence of a pathogenic strain isolated from blood cultures of a patient with infectious endocarditis. PMID:27660783

  14. Susceptibility of mixed infection of Ureaplasma Urealyticum and Mycoplasma Hominis to seven antimicrobial agents and comparison with that of Ureaplasma Urealyticum infection.

    PubMed

    Huang, Changzheng; Liu, Zhixiang; Lin, Nengxing; Tu, Yating; Li, Jiawen; Zhang, Demei

    2003-01-01

    In order to investigate the susceptibility of mixed infection of Ureaplasma Urealyticum (UU) and Mycoplasma Hominis (MH) to 7 kinds of antimicrobial agents and comparison with that of UU infection in NGU patients, the in vitro susceptibility was determined by using microdilution method. The positive results were analyzed. The results showed that the sequence of susceptibility to 7 kinds of antimicrobial agents for both UU infection group and UU-MH mixed infection group was almost the same from the highest susceptibility to the lowest accordingly: Josamycin, Doxycycline, Minocycline, Sparfloxacin, Roxithromycin, Ofloxacin and Azithromycin. The total drug resistance rate for UU-MH mixed infection group (97.67%) was significantly higher than that for UU infection group (44.67%, P < 0.01). The highest drug resistance rate in UU group and UU-MH mixed infection group was 31.33% (Ofloxacin) and 90.48% (Azithromycin) respectively. UU-MH mixed infection showed an increased drug resistance and changes of drug resistance spectrum.

  15. Comparative Genomic Analyses of Attenuated Strains of Mycoplasma gallisepticum▿ †

    PubMed Central

    Szczepanek, S. M.; Tulman, E. R.; Gorton, T. S.; Liao, X.; Lu, Z.; Zinski, J.; Aziz, F.; Frasca, S.; Kutish, G. F.; Geary, S. J.

    2010-01-01

    Mycoplasma gallisepticum is a significant respiratory and reproductive pathogen of domestic poultry. While the complete genomic sequence of the virulent, low-passage M. gallisepticum strain R (Rlow) has been reported, genomic determinants responsible for differences in virulence and host range remain to be completely identified. Here, we utilize genome sequencing and microarray-based comparative genomic data to identify these genomic determinants of virulence and to elucidate genomic variability among strains of M. gallisepticum. Analysis of the high-passage, attenuated derivative of Rlow, Rhigh, indicated that relatively few total genomic changes (64 loci) occurred, yet they are potentially responsible for the observed attenuation of this strain. In addition to previously characterized mutations in cytadherence-related proteins, changes included those in coding sequences of genes involved in sugar metabolism. Analyses of the genome of the M. gallisepticum vaccine strain F revealed numerous differences relative to strain R, including a highly divergent complement of vlhA surface lipoprotein genes, and at least 16 genes absent or significantly fragmented relative to strain R. Notably, an Rlow isogenic mutant in one of these genes (MGA_1107) caused significantly fewer severe tracheal lesions in the natural host compared to virulent M. gallisepticum Rlow. Comparative genomic hybridizations indicated few genetic loci commonly affected in F and vaccine strains ts-11 and 6/85, which would correlate with proteins affecting strain R virulence. Together, these data provide novel insights into inter- and intrastrain M. gallisepticum genomic variability and the genetic basis of M. gallisepticum virulence. PMID:20123709

  16. Variant colony surface antigenic phenotypes within mycoplasma strain populations: implications for species identification and strain standardization.

    PubMed Central

    Rosengarten, R; Yogev, D

    1996-01-01

    Immunobinding assays with mycoplasma colonies on agar plates (immunofluorescence and immunoperoxidase techniques) or with imprints of colonies transferred to solid supports (colony immunoblotting) are widely used as standard diagnostic tests for serological species identification of mycoplasma isolates. However, in light of the high rate of variability of surface antigens in many mycoplasmas, diagnostic data obtained with these techniques require a more critical evaluation. In this report, we demonstrate with some examples that mycoplasma surface variability based on alterations in expression, in size, and in surface presentation of integral and peripheral membrane proteins may lead to misinterpretation of colony immunostaining reactions obtained by using specific monoclonal antibodies as well as conventional diagnostic hyperimmune sera. To more easily identify phenotypically mixed isolates or samples which contain more than one species, we have introduced some minor modifications of the colony immunoblot technique which provide sharp signals of positive as well as negative reactions and enable identification of cryptic epitopes. It is further demonstrated that because of the variability in colony surface antigenic phenotype, mycoplasma strains, including well-established reference and other prototype strains which are used under the same designation in many laboratories, can differ markedly in their antigen profiles and their potentially virulence-related surface properties, since they are usually purified by filter cloning and often propagated by subcultivation of randomly selected agar-grown subpopulations. We conclude from this study that because of this surface variability, the establishment of criteria for standardization of mycoplasma strains and diagnostic antisera is urgently required in order to obtain reproducible results in different laboratories. PMID:8748292

  17. 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...

  18. Genome Sequences of Two Tunisian Field Strains of Avian Mycoplasma, M. meleagridis and M. gallinarum

    PubMed Central

    Yacoub, Elhem; Sirand-Pugnet, Pascal; Barré, Aurélien; Blanchard, Alain; Hubert, Christophe; Maurier, Florence; Bouilhol, Emmanuel

    2016-01-01

    Mycoplasma meleagridis and Mycoplasma gallinarum are bacteria that affect birds, but little is known about the genetic basis of their interaction with chickens and other poultry. Here, we sequenced the genomes of M. meleagridis strain MM_26B8_IPT and M. gallinarum strain Mgn_IPT, both isolated from chickens showing respiratory symptoms, poor growth, reduction in hatchability, and loss of production. PMID:27313300

  19. Genome Sequences of Two Tunisian Field Strains of Avian Mycoplasma, M. meleagridis and M. gallinarum.

    PubMed

    Yacoub, Elhem; Sirand-Pugnet, Pascal; Barré, Aurélien; Blanchard, Alain; Hubert, Christophe; Maurier, Florence; Bouilhol, Emmanuel; Ben Abdelmoumen Mardassi, Boutheina

    2016-01-01

    Mycoplasma meleagridis and Mycoplasma gallinarum are bacteria that affect birds, but little is known about the genetic basis of their interaction with chickens and other poultry. Here, we sequenced the genomes of M. meleagridis strain MM_26B8_IPT and M. gallinarum strain Mgn_IPT, both isolated from chickens showing respiratory symptoms, poor growth, reduction in hatchability, and loss of production. PMID:27313300

  20. 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

  1. Observations on Membranes of Mycoplasma laidlawii Strain B

    PubMed Central

    Smith, P. F.; Koostra, W. L.; Mayberry, W. R.

    1969-01-01

    The cytoplasmic membrane of Mycoplasma laidlawii strain B is solubilized by anionic and nonionic detergents, succinylation, phospholipase A, alkaline phosphatase, trypsin, and chymotrypsin. Cationic detergents are without effect, as are chelating agents, even in the presence of high concentrations of monovalent cation. The detergent-solubilized membrane exhibits one peak in the analytical ultracentrifuge, but the sedimentation coefficient is dependent upon concentration of detergent. Simple dialysis does not remove all of the sodium dodecylsulfate except from lipid-depleted membrane particles. Membranes bind sodium dodecylsulfate but acetone powders of membranes do not. Sulfated alcohols with chain lengths of C14 and C16 are more tightly bound than dodecylsulfate. A constant amount of di- and trivalent cation is bound by the membrane upon aggregation. Only a portion of this cation is removable with chelating agents. No chelating agent is bound by these aggregates. A portion of the lipid-depleted membrane particles is solubilized by negatively charged lipids and detergents, giving rise to aggregates in the presence of divalent cation. Fractionations of detergent-solubilized membranes by preparative gel electrophoresis and ammonium sulfate were inconclusive. Density gradient centrifugation of succinylated membranes yielded at least five fractions which exhibited homogeneity by ultracentrifugation. Analytical gel electrophoresis of these fractions demonstrated heterogeneity. The composition of these five fractions suggested separation of protein from lipid. PMID:5361209

  2. Complete Genome Sequence of Mycoplasma bovoculi Strain M165/69T (ATCC 29104)

    PubMed Central

    Foecking, Mark F.

    2014-01-01

    Bovine ocular infections compromise animal health and result in significant economic losses. Mycoplasma bovoculi is an etiological agent of conjunctivitis. Presented here is the 760,240-bp complete genome sequence of the M. bovoculi type strain M165/69T. An analysis of the deduced proteome provides insights into the adherence and antigenic variation mechanisms of the strain. PMID:24558249

  3. A comparison of adherence by four strains of Staphylococcus intermedius and Staphylococcus hominis to canine corneocytes collected from normal dogs and dogs suffering from atopic dermatitis.

    PubMed

    McEwan, N A; Kalna, G; Mellor, D

    2005-06-01

    The aim of this study was to compare the adherence of four strains of Staphylococcus intermedius and a single strain of Staphylococcus hominis to corneocytes from both normal dogs and dogs suffering from atopic dermatitis. Cells from the skin surface, corneocytes, were collected from 10 normal dogs and 10 dogs suffering from atopic dermatitis. Four strains of S. intermedius, three isolated from canine pyoderma skin lesions (strains A, B and C), and one isolated form from canine synovial membrane sample from a case of septic arthritis (strain D) were compared. S. hominis, which is not normally associated with canine disease, was also evaluated for its ability to adhere to canine corneocytes. S. hominis did not adhere to canine corneocytes. All four strains of S. intermedius adhered well to canine corneocytes collected from both normal and atopic dogs. All strains of S. intermedius showed statistically greater adherence to corneocytes collected from atopic dogs compared with those collected from normal dogs. It was concluded that the adherence assay employed here showed that S. hominis does not adhere to canine corneocytes, S. intermedius adheres preferentially to atopic corneocytes.

  4. Collaborative study report: evaluation of the ATCC experimental mycoplasma reference strains panel prepared for comparison of NAT-based and conventional mycoplasma detection methods.

    PubMed

    Dabrazhynetskaya, Alena; Volokhov, Dmitriy V; Lin, Tsai-Lien; Beck, Brian; Gupta, Rajesh K; Chizhikov, Vladimir

    2013-11-01

    The main goal of this collaborative study was to evaluate the experimental panel of cryopreserved mycoplasma reference strains recently prepared by the American Type Culture Collection (ATCC(®)) in order to assess the viability and dispersion of cells in the mycoplasma stocks by measuring the ratio between the number of genomic copies (GC) and the number of colony forming units (CFU) in the reference preparations. The employment of microbial reference cultures with low GC/CFU ratios is critical for unbiased and reliable comparison of mycoplasma testing methods based on different methodological approaches, i.e., Nucleic Acid Testing (NAT) and compendial culture-based techniques. The experimental panel included ten different mycoplasma species known to represent potential human and animal pathogens as well as common contaminants of mammalian and avian cell substrates used in research, development, and manufacture of biological products. Fifteen laboratories with expertise in field of mycoplasma titration and quantification of mycoplasmal genomic DNA participated in the study conducted from February to October of 2012. The results of this study demonstrated the feasibility of preparing highly viable and dispersed (possessing low GC/CFU ratios) frozen stocks of mycoplasma reference materials, required for reliable comparison of NAT-based and conventional mycoplasma detection methods.

  5. Genome Sequence of Mycoplasma ovipneumoniae Strain SC01 ▿

    PubMed Central

    Yang, Falong; Tang, Cheng; Wang, Yong; Zhang, Huanrong; Yue, Hua

    2011-01-01

    Mycoplasma ovipneumoniae is associated with chronic nonprogressive pneumonia in both sheep and goats. Studies concerning its molecular pathogenesis, genetic analysis, and vaccine development have been hindered due to limited genomic information. Here, we announce the first complete genome sequence of this organism. PMID:21742877

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

    PubMed

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

    1988-02-01

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

  7. Complete Genome Sequence of Mycoplasma mycoides subsp. mycoides T1/44, a Vaccine Strain against Contagious Bovine Pleuropneumonia.

    PubMed

    Gourgues, Géraldine; Barré, Aurélien; Beaudoing, Emmanuel; Weber, Johann; Magdelenat, Ghislaine; Barbe, Valérie; Schieck, Elise; Jores, Joerg; Vashee, Sanjay; Blanchard, Alain; Lartigue, Carole; Sirand-Pugnet, Pascal

    2016-01-01

    Mycoplasma mycoidessubsp.mycoidesis the etiologic agent of contagious bovine pleuropneumonia. We report here the complete genome sequence of the strain T1/44, which is widely used as a live vaccine in Africa. PMID:27081135

  8. Complete Genome Sequence of Mycoplasma mycoides subsp. mycoides T1/44, a Vaccine Strain against Contagious Bovine Pleuropneumonia

    PubMed Central

    Gourgues, Géraldine; Barré, Aurélien; Beaudoing, Emmanuel; Weber, Johann; Magdelenat, Ghislaine; Barbe, Valérie; Schieck, Elise; Jores, Joerg; Vashee, Sanjay; Blanchard, Alain; Lartigue, Carole

    2016-01-01

    Mycoplasma mycoides subsp. mycoides is the etiologic agent of contagious bovine pleuropneumonia. We report here the complete genome sequence of the strain T1/44, which is widely used as a live vaccine in Africa. PMID:27081135

  9. A sequence variant of Staphylococcus hominis with a high prevalence of oxacillin and fluoroquinolone resistance.

    PubMed

    Fitzgibbon, J E; Nahvi, M D; Dubin, D T; John, J F

    2001-11-01

    A newly identified subspecies of Staphylococcus hominis, S. hominis subsp. novobiosepticus, was found to be the cause of several invasive infections at a hospital in New Jersey. This subspecies differs from classical S. hominis, now S. hominis subsp. hominis, by the phenotypic characteristics of novobiocin resistance and the inability to ferment trehalose. DNA sequences of segments of 16S rRNA, DNA gyrase (gyrA), and DNA topoisomerase IV (grlA) genes were determined for the type strains of the 2 subspecies, and for 34 S. hominis clinical isolates. The 16S rRNA sequences of the type strains differed at 3 positions over 410 bp; the grlA sequences differed at 6 positions over 119 bp. These sequence differences define S. hominis subsp. novobiosepticus and S. hominis subsp. hominis "sequevars." Of 34 S. hominis clinical isolates, 31 were S. hominis subsp. novobiosepticus sequevars, 28 of which were resistant to both oxacillin and ciprofloxacin. The clinical microbiology laboratory, using a MicroScan system, identified 7 of the 31S. hominis subsp. novobiosepticus sequevars as S. hominis subsp. hominis on the basis of phenotypic characteristics. Three S. hominis subsp. hominis sequevars were all identified phenotypically as S. hominis subsp. hominis and were oxacillin- and ciprofloxacin-susceptible. Although the precise relationship between the S. hominis sequevars and their phenotypic subspecies remains to be determined, our results indicate that antibiotic-resistant clinical isolates of S. hominis belong almost exclusively to the S. hominis subsp. novobiosepticus sequevar.

  10. Comparison of Mycoplasma arthritidis strains by enzyme-linked immunosorbent assay, immunoblotting, and DNA restriction analysis.

    PubMed Central

    Washburn, L R; Voelker, L L; Ehle, L J; Hirsch, S; Dutenhofer, C; Olson, K; Beck, B

    1995-01-01

    Twenty Mycoplasma arthritidis strains or isolates were compared by a combination of enzyme-linked immunosorbent assay by an antiserum adsorption technique, Western immunoblotting, and restriction analysis of chromosomal DNA. Antigenic markers that defined strains related to strains 158p10p9, PG6, and H606 were identified. In addition, restriction analysis allowed all 20 strains to be divided into six groups. Results of restriction analysis corresponded generally with antigenic similarities, although the former did not allow grouping with as fine a precision as the latter. However, intrastrain antigenic variability, which is common among many Mycoplasma species, including M. arthritidis, introduced a complicating factor into our attempts at antigenic analysis. While serologic and antigenic analyses remain useful, we recommend that they be used with caution and in combination with other techniques for identifying and characterizing new isolates and newly acquired strains. Combinations of these techniques have proven to be useful in our laboratory for quality control and for uncovering interesting relationships among strains subjected to animal passage and their less virulent antecedents and among strains originally classified as the same but obtained from different sources and maintained, sometimes for decades, in different laboratories. PMID:7494014

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

    PubMed

    Ferguson-Noel, N M; Williams, S M

    2015-01-01

    The efficacy of a live Mycoplasma gallisepticum (MG) vaccine candidate (K-strain) was compared to commercially available vaccines in broiler-type chickens (Trial 1) and layer-type chickens (Trial 2). In Trial 1, three-week-old broiler-type chickens were vaccinated via aerosol with K-strain or an F-strain vaccine. The vaccinated chickens and 10 non-vaccinated controls were subsequently challenged with virulent R-strain via aerosol at six weeks post vaccination; both K-strain and F-strain vaccination resulted in significant protection from air sac and tracheal lesions, as well as R-strain colonization (P ≤ 0.05). In Trial 2, commercial layer-type chickens were vaccinated with ts-11 (via eye drop) or K-strain (via aerosol) at 12 weeks of age. At 25 weeks of age these birds were challenged with R-strain via aerosol. The ts-11 and K-strain vaccinated groups both had significantly lower air sac lesion scores and a lower prevalence of ovarian regression after challenge as compared to non-vaccinated chickens (P ≤ 0.05). K-strain vaccination also prevented significant tracheal lesions and R-strain colonization (P ≤ 0.05). K-strain shows great potential as a highly efficacious live MG vaccine in broiler and layer-type chickens for protection of the respiratory and reproductive systems as well as prevention of infection with field strains. PMID:25571953

  12. Bacteremia by Dermabacter hominis, a Rare Pathogen

    PubMed Central

    Gómez-Garcés, José Luis; Oteo, Jesús; García, Guadalupe; Aracil, Belén; Alós, Juan Ignacio; Funke, Guido

    2001-01-01

    Dermabacter hominis is a gram-positive, catalase-positive, glucose-fermenting rod, which, as it grows forms small greyish-white colonies with a characteristic pungent odor. Previously known as coryneform Centers for Disease Control and Prevention groups 3 and 5, it was catalogued as D. hominis in 1994. Various strains isolated in blood cultures, abscesses, or wounds in the 1970s were retrospectively characterized in referral centers as D. hominis. In this report we describe two patients with severe underlying pathology who developed bacteremias by D. hominis within the context of their clinical pictures. PMID:11376092

  13. Genome Anatomy of Pyrenochaeta unguis-hominis UM 256, a Multidrug Resistant Strain Isolated from Skin Scraping.

    PubMed

    Toh, Yue Fen; Yew, Su Mei; Chan, Chai Ling; Na, Shiang Ling; Lee, Kok Wei; Hoh, Chee-Choong; Yee, Wai-Yan; Ng, Kee Peng; Kuan, Chee Sian

    2016-01-01

    Pyrenochaeta unguis-hominis is a rare human pathogen that causes infection in human skin and nail. P. unguis-hominis has received little attention, and thus, the basic biology and pathogenicity of this fungus is not fully understood. In this study, we performed in-depth analysis of the P. unguis-hominis UM 256 genome that was isolated from the skin scraping of a dermatitis patient. The isolate was identified to species level using a comprehensive multilocus phylogenetic analysis of the genus Pyrenochaeta. The assembled UM 256 genome has a size of 35.5 Mb and encodes 12,545 putative genes, and 0.34% of the assembled genome is predicted transposable elements. Its genomic features propose that the fungus is a heterothallic fungus that encodes a wide array of plant cell wall degrading enzymes, peptidases, and secondary metabolite biosynthetic enzymes. Antifungal drug resistance genes including MDR, CDR, and ERG11/CYP51 were identified in P. unguis-hominis UM 256, which may confer resistance to this fungus. The genome analysis of P. unguis-hominis provides an insight into molecular and genetic basis of the fungal lifestyles, understanding the unrevealed biology of antifungal resistance in this fungus. PMID:27626635

  14. Genome Anatomy of Pyrenochaeta unguis-hominis UM 256, a Multidrug Resistant Strain Isolated from Skin Scraping

    PubMed Central

    Toh, Yue Fen; Yew, Su Mei; Chan, Chai Ling; Na, Shiang Ling; Lee, Kok Wei; Hoh, Chee-Choong; Yee, Wai-Yan; Ng, Kee Peng

    2016-01-01

    Pyrenochaeta unguis-hominis is a rare human pathogen that causes infection in human skin and nail. P. unguis-hominis has received little attention, and thus, the basic biology and pathogenicity of this fungus is not fully understood. In this study, we performed in-depth analysis of the P. unguis-hominis UM 256 genome that was isolated from the skin scraping of a dermatitis patient. The isolate was identified to species level using a comprehensive multilocus phylogenetic analysis of the genus Pyrenochaeta. The assembled UM 256 genome has a size of 35.5 Mb and encodes 12,545 putative genes, and 0.34% of the assembled genome is predicted transposable elements. Its genomic features propose that the fungus is a heterothallic fungus that encodes a wide array of plant cell wall degrading enzymes, peptidases, and secondary metabolite biosynthetic enzymes. Antifungal drug resistance genes including MDR, CDR, and ERG11/CYP51 were identified in P. unguis-hominis UM 256, which may confer resistance to this fungus. The genome analysis of P. unguis-hominis provides an insight into molecular and genetic basis of the fungal lifestyles, understanding the unrevealed biology of antifungal resistance in this fungus. PMID:27626635

  15. Integrative conjugative elements are widespread in field isolates of Mycoplasma species pathogenic for ruminants.

    PubMed

    Tardy, Florence; Mick, Virginie; Dordet-Frisoni, Emilie; Marenda, Marc Serge; Sirand-Pugnet, Pascal; Blanchard, Alain; Citti, Christine

    2015-03-01

    Comparative genomics have revealed massive horizontal gene transfer (HGT) between Mycoplasma species sharing common ruminant hosts. Further results pointed toward an integrative conjugative element (ICE) as an important contributor of HGT in the small-ruminant-pathogen Mycoplasma agalactiae. To estimate the prevalence of ICEs in ruminant mycoplasmas, we surveyed their occurrence in a collection of 166 field strains representing 4 (sub)species that are recognized as major pathogens. Based on available sequenced genomes, we first defined the conserved, minimal ICE backbone as composed of 4 coding sequences (CDSs) that are evenly distributed and predicted to be essential for ICE chromosomal integration-excision and horizontal transfer. Screening of the strain collection revealed that these 4 CDSs are well represented in ruminant Mycoplasma species, suggesting widespread occurrence of ICEs. Yet their prevalence varies within and among species, with no correlation found with the individual strain history. Extrachromosomal ICE forms were also often detected, suggesting that ICEs are able to circularize in all species, a first and essential step in ICE horizontal transfer. Examination of the junction of the circular forms and comparative sequence analysis of conserved CDSs clearly pointed toward two types of ICE, the hominis and spiroplasma types, most likely differing in their mechanism of excision-integration. Overall, our data indicate the occurrence and maintenance of functional ICEs in a large number of field isolates of ruminant mycoplasmas. These may contribute to genome plasticity and gene exchanges and, presumably, to the emergence of diverse genotypes within pathogenic mycoplasmas of veterinary importance.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  18. A case of septic arthritis caused by a Mycoplasma salivarium strain resistant towards Ciprofloxacin and Clarithromycin in a patient with chronic lymphatic leukemia.

    PubMed

    Büchsel, Martin; Pletschen, Lars; Fleiner, Michael; Häcker, Georg; Serr, Annerose

    2016-09-01

    Mycoplasma salivarium is a rare agent of septic arthritis in immunocompromised patients. We report a case of septic arthritis due to Mycoplasma salivarium in a patient with B-cell chronic lymphocytic leukemia who underwent chemotherapy with rituximab and bendamustin. Therapy of arthritis due to Mycoplasma salivarium is difficult because there are almost no susceptibility data available. The present case illustrates that antimicrobial susceptibility of Mycoplasma strains is not necessarily predictable and that antibiotic therapy should therefore be guided by in vitro susceptibility testing. PMID:27342785

  19. A case of septic arthritis caused by a Mycoplasma salivarium strain resistant towards Ciprofloxacin and Clarithromycin in a patient with chronic lymphatic leukemia.

    PubMed

    Büchsel, Martin; Pletschen, Lars; Fleiner, Michael; Häcker, Georg; Serr, Annerose

    2016-09-01

    Mycoplasma salivarium is a rare agent of septic arthritis in immunocompromised patients. We report a case of septic arthritis due to Mycoplasma salivarium in a patient with B-cell chronic lymphocytic leukemia who underwent chemotherapy with rituximab and bendamustin. Therapy of arthritis due to Mycoplasma salivarium is difficult because there are almost no susceptibility data available. The present case illustrates that antimicrobial susceptibility of Mycoplasma strains is not necessarily predictable and that antibiotic therapy should therefore be guided by in vitro susceptibility testing.

  20. Nosocomial spread of a Staphylococcus hominis subsp. novobiosepticus strain causing sepsis in a neonatal intensive care unit.

    PubMed

    Chaves, Fernando; García-Alvarez, Mónica; Sanz, Francisca; Alba, Concepción; Otero, Joaquín R

    2005-09-01

    From 2002 to 2003, 32 isolates of Staphylococcus hominis subsp. novobiosepticus (SHN) were recovered from 21 patients, 18 of whom were neonates, with 13 considered to have late-onset SHN sepsis. All isolates from neonates had an indistinguishable pulsed-field gel electrophoresis pattern. Our data support SHN as an important nosocomial pathogen in neonates.

  1. Changes in pathogenicity and immunogenicity of Mycoplasma mycoides subsp. mycoides strains revealed by comparative genomics analysis

    PubMed Central

    Li, Yuan; Wang, Yang; Wang, Rui; Zhu, Yongqiang; Liu, Suli; Wang, Qi; Shao, Jiari; Chen, Ying; Gao, Liping; Zhou, Changping; Liu, Henggui; Wang, Xiumei; Zheng, Huajun; Xin, Jiuqing

    2016-01-01

    Mycoplasma mycoides subsp. mycoides is the causative agent of contagious bovine pleuropneumonia. A pathogenic strain BEN-1 was isolated from bovine lung and underwent continuous passages in rabbits for 468 generations. During this process, the strain’s strong virulence became weak and, gradually, it lost the ability to confer protective immunity in cattle but developed virulence in rabbits. In order to gain insight into the mechanisms behind the reduction in virulence and the loss of immunogenicity, we sequenced five representative strains of the BEN series, including the original strain (BEN-1), the strain generation that first acquired virulence in rabbits (BEN-50), the two vaccine strain generations (BEN-181 and BEN-326), and the strain generation showing the greatest loss of immunogenicity (BEN-468). The gene mutation rate in the four different propagation stages varied greatly, and over half of variations observed in each generation were removed during the propagation process. However, the variation maintained in the BEN-468 generation might contribute to its changes in virulence and immunogenicity. We thus identified 18 genes associated with host adaptation, six genes contributing to virulence in cattle, and 35 genes participating in conferring immunity in cattle. These findings might help us optimize the vaccine to obtain more effective immunization results. PMID:26750304

  2. Incidence and antibiotic susceptibility of genital mycoplasmas in sexually active individuals in Hungary.

    PubMed

    Pónyai, K; Mihalik, N; Ostorházi, E; Farkas, B; Párducz, L; Marschalkó, M; Kárpáti, S; Rozgonyi, F

    2013-11-01

    The aim of this study was to examine the incidence and antibiotic sensitivity of Ureaplasma urealyticum and Mycoplasma hominis strains cultured from the genital discharges of sexually active individuals who attended our STD outpatient service. Samples were taken with universal swab (Biolab®, Budapest, Hungary) into the Urea-Myco DUO kit (Bio-Rad®, Budapest, Hungary) and incubated in ambient air for 48 h at 37 °C. The determination of antibiotic sensitivity was performed in U9 and arginin broth using the SIR Mycoplasma kit (Bio-Rad®, Budapest, Hungary) under the same conditions. Between 01.05.2008 and 31.12.2011, 373/4,466 (8.35 %) genito-urethral samples with U. urealyticum and 41/4,466 (0.91 %) genito-urethral samples with M. hominis infection were diagnosed in sexually active individuals in the National STD Center, Semmelweis University. U. urealyticum was isolated in 12.54 % in the cervix and 4.1 % in the male urethra, while M. hominis was isolated in 1.33 % in the cervix and 0.51 % in the male urethra. The affected age group was between 21 and 60 years old. U. urealyticum strains were sensitive to tetracycline (95.9 %), doxycycline (97.32 %), and azithromycin (85.79 %), and resistant to erythromycin (81.23 %), clindamycin (75.06 %), and ofloxacin (25.2 %). Cross-resistance occurred in 38.71 % of patients to erythromycin and clindamycin. M. hominis strains were sensitive to clindamycin, ofloxacin, and doxycycline in more than 95 %, to tetracycline in 82.92 %, and no cross-resistance was detected among the antibiotics. Our study confirms that the continuously changing antibiotic resistance of ureaplasmas and mycoplasmas should be followed at least in a few centers in every country, so as to determine the best local therapy options for sexually transmitted infection (STI) patients.

  3. Evaluation of Mycoplasma gallisepticum K-strain as a live vaccine in chickens.

    PubMed

    Ferguson-Noel, N M; Laibinis, V A; Kleven, S H

    2012-03-01

    We evaluated the pathogenicity of three live Mycoplasma gallisepticum (MG) vaccine candidates by infection via aerosol of 3-wk-old chickens with log phase broth cultures (trial 1). Two of the candidates (K3020 and K4649A) colonized only 10% and 20% of the chickens, respectively, unlike K2101 (K-strain), which was reisolated from all of the vaccinated chickens tested. K-strain inoculation did not result in significant air sac or tracheal lesions in chickens at 10 and 39 days postinfection (P < or = 0.05). The efficacy of K-strain as a live vaccine was evaluated in trial 2, by challenge of vaccinated chickens with virulent R-strain via aerosol at 6 wk postvaccination. K-strain vaccination resulted in significant protection from air sac and tracheal lesions (P < or = 0.05). The K-strain was further investigated to evaluate transmissibility (trial 3), colonization and persistence of infection following aerosol administration (trial 4), genetic and phenotypic stability following back passage through chickens (trial 5), and vertical transmission (trial 6). The K-strain had a low rate of horizontal transmission; it remained primarily in the respiratory system of inoculated birds and persisted in the upper respiratory tract for the duration of the trial 4 (5 mo). There was no increase in virulence of K-strain when it was back passaged five times through chickens, and no vertical transmission of K-strain was detected. K-strain showed great potential as a safe and effective live MG vaccine. PMID:22545527

  4. Colonisation of the respiratory tract of lambs by strains of Mycoplasma ovipneumoniae.

    PubMed

    Ionas, G; Mew, A J; Alley, M R; Clarke, J K; Robinson, A J; Marshall, R B

    1985-12-01

    The age and time of year when colonisation of the nasal cavity of lambs by Mycoplasma ovipneumoniae occurs; the persistence of the organism, and its prevalence in the lungs at slaughter were examined in 2 flocks of sheep in New Zealand. No colonisation had occurred at the time of weaning at 6-7 weeks, but M. ovipneumoniae was recovered from most lambs on at least one occasion before they were slaughtered when about 8 months old. In most cases, colonisation of the nasal cavity by M. ovipneumoniae was a transient phenomenon. At slaughter M. ovipneumoniae was recovered from the lungs of 89% of the lambs of one flock and 80% of the other flock. Bacterial restriction endonuclease DNA analysis (BRENDA) of 34 nasal isolates from one flock showed that it was possible to identify 7 "groups" each with markedly different BRENDA patterns. Lambs initially colonised by one strain, often lost that strain, and if recolonisation occurred it was with a different strain. M. ovipneumoniae was recovered at slaughter from the lungs of most lambs, both normal and pneumonic. The isolates from one flock were examined by BRENDA, and approximately 90% of them gave similar or identical patterns. The predominant strain isolated from the lungs had been recovered from the nasal cavity of many of the lambs about 3 weeks earlier. This suggests that the nasal and lung isolates do not represent independent populations. However, nasal strains may differ in their ability to colonise the lungs.(ABSTRACT TRUNCATED AT 250 WORDS)

  5. Emergence of atypical Mycoplasma agalactiae strains harboring a new prophage and associated with an alpine wild ungulate mortality episode.

    PubMed

    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; Citti, Christine

    2012-07-01

    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. Emergence of atypical Mycoplasma agalactiae strains harboring a new prophage and associated with an alpine wild ungulate mortality episode.

    PubMed

    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; Citti, Christine

    2012-07-01

    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.

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

    PubMed

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

    2003-02-22

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

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

    PubMed

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

    2003-02-22

    Two cases of Mycoplasma gallisepticum infection in different avian species in backyard gamebird operations in Slovenia were investigated. In the first case, M gallisepticum was associated with severe respiratory disease with almost 20 per cent mortality in pheasants, whereas the infection was less pathogenic for chickens and turkeys reared at the same site. The M gallisepticum isolates from pheasants had a unique pMGA gene sequence containing a repeat of 12 nucleotides, and they contained only small amounts of the cytadhesins MGC1 and MGC3 and no PvpA protein. However, they expressed some typical M gallisepticum proteins and several proteins which were immunogenic for pheasants, chickens and turkeys. A strain of M gallisepticum isolated from the sinus of a pheasant was highly pathogenic for chicken embryos. In the second case, the M gallisepticum strain that was associated with respiratory disease and mortality in peafowl also affected chickens. M gallisepticum strain ULB 992 was isolated from the infraorbital sinus of a dead peafowl. The ULB 992 strain synthesised a small amount of MGC3, a truncated form of MGC1 and lacked PvpA. However, it expressed several proteins which were immunogenic for the birds infected with M gallisepticum at both gamebird operations. PMID:12625537

  9. The isolation of multiple strains of Mycoplasma ovipneumoniae from individual pneumonic sheep lungs.

    PubMed

    Ionas, G; Clarke, J K; Marshall, R B

    1991-11-01

    The heterogeneity of Mycoplasma ovipneumoniae isolates from the lungs of sheep with chronic non-progressive pneumonia (CNP) from the same flock raised the possibility that multiple isolates derived from one lung were not all identical. To test this hypothesis, thirty isolates were obtained from each of six pneumonic sheep lungs at slaughter. Four lungs had relatively severe lesions and from each of these, three or four strains of M. ovipneumonia, distinguishable by REA and in most cases by SDS-PAGE, were detected. From the lungs of each of two sheep with mild lesions, two strains of M. ovipneumoniae were detected. Four isolates from one lung were further examined by restriction endonuclease analysis (REA) using many restriction endonucleases. Those which differed with EcoRI also differed when other restriction endonucleases were used. However, partial digests occurred mainly with those restriction endonucleases which recognise cytosine-rich sequences. The presence of multiple strains of one species of microorganism in individual lesions is an unusual concept which may not be limited to one disease or to one host.

  10. Herd-specific strains of Mycoplasma bovis in outbreaks of mycoplasmal mastitis and pneumonia.

    PubMed

    Aebi, Marlis; Bodmer, Michèle; Frey, Joachim; Pilo, Paola

    2012-06-15

    Mycoplasma bovis causes severe economic losses in livestock production, particularly on the Northern American continent and more recently also in continental Europe. The aim of the current study was to evaluate whether the recently emerging outbreaks were due to a particular clone or strain of M. bovis or whether these outbreaks are due to multiple infectious strains of M. bovis. The study is based on the analysis M. bovis isolated from cattle of herds with outbreaks of mycoplasmal mastitis or pneumonia from geographically non related parts of Switzerland. M. bovis isolates were typed by insertion sequence (IS) element analysis based upon ISMbov1 and ISMbov2 southern-blot hybridization. We observed a strong divergence of M. bovis strains among affected herds which mostly were herd specific. This argues against the assumption that a recent infiltration of a particular clone of M. bovis is the cause of the perilous emerging outbreaks. The study suggests that transmission occurs from animal to animal most probably via milk. PMID:22306036

  11. Comparison of Mycoplasma gallisepticum strains and identification of immunogenic integral membrane proteins with Triton X-114 by immunoblotting.

    PubMed

    Avakian, A P; Kleven, S H; Ley, D H

    1991-11-01

    Pooled chicken antisera from 33 and 77 days post Mycoplasma gallisepticum strain R contact-exposure reacted with cell proteins of 19 M. gallisepticum strains. These pooled antisera reacted with more proteins and with greater intensity to reference strains (R, PG31, S6, and A5969) and nine field strains than they did with six other field strains including three (703, 503, and 730) that have been described as serological variants. Following extraction with Triton X-114 the majority of immunogenic M. gallisepticum proteins partitioned exclusively or primarily into the detergent phase indicating that they are integral membrane proteins. This included three immunogenic species-specific proteins (p64, p56 and p26). M. gallisepticum p56 was detected, by immunoblotting, in 18 of 19 strains suggesting that it could serve as an antigen for serological tests. P26 was evident in 13 of 19 strains. Hyperimmune antiserum to p64 reacted with a 64 kDa protein in 19 M. gallisepticum strains, but did not react with seven other avian Mycoplasma spp. There was no evidence found supporting the view that p64 is the hemagglutinin of M. gallisepticum.

  12. Effects of time specific F-strain Mycoplasma gallisepticum inoculation overlays on pre-lay ts11-strain Mycoplasma gallisepticum inoculation on performance characteristics of commercial laying hens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mycoplasma bacteria are virtually ubiquitous in layer chicken flocks and M. gallisepticum is the species of greatest concern to commercial egg producers. Live M. gallisepticum vaccines were initially approved by the USDA for use in commercial layers in 1988 to help control M. gallisepticum outbreaks...

  13. Effect of Diethylaminoethyl Dextran on the Growth of Mycoplasma in Agar

    PubMed Central

    Tauraso, Nicola M.

    1967-01-01

    The growth of certain strains of Mycoplasma is inhibited by substances present in commercial agar preparations. The addition of diethylaminoethyl (DEAE) dextran (10 mg per 100 ml) to agar media appears to enhance the growth of some strains. Of eight strains initially tested, the presence of DEAE dextran grossly enhanced the growth of three strains. One strain appeared not to be affected, and a clearly enhancing effect was not evident with four strains. Quantitative studies revealed that growth enhancement varied from 10 colony-forming units (CFU) for M. hominis type II (strain Campo) to 103.3 CFU for M. pulmonis (strain 880). The growth-enhancing effect is probably due to the ability of DEAE dextran to bind the sulfated polysaccharide moieties in agar and not to the DEAE dextran, per se. Images PMID:6025444

  14. Characterization of P40, a Cytadhesin of Mycoplasma agalactiae

    PubMed Central

    Fleury, Bénédicte; Bergonier, Dominique; Berthelot, Xavier; Peterhans, Ernst; Frey, Joachim; Vilei, Edy M.

    2002-01-01

    An immunodominant protein, P40, of Mycoplasma agalactiae was analyzed genetically and functionally. The gene encoding P40 was cloned from type strain PG2, sequenced, submitted to point mutagenesis in order to convert mycoplasma-specific TGATrp codon to the universal TGGTrp codon, and subsequently expressed in Escherichia coli. Nucleotide sequence-derived amino acid sequence comparisons revealed a similarity of P40 to the adhesin P50 of Mycoplasma hominis and to protein P89 of Spiroplasma citri, which is expected to be involved in adhesion. The amino acid sequence of P40 revealed a recognition site for a signal peptidase and strong antigenic and hydrophilic motifs in the C-terminal domain. Triton X-114 phase partitioning confirmed that P40 is a membrane protein. Fab fragments of antibodies directed against recombinant purified P40 significantly inhibited adherence of M. agalactiae strains PG2 to lamb joint synovial cells LSM 192. Sera taken sequentially from sheep infected with PG2 revealed that P40 induced a strong and persistent immune response that gave strong signals on immunoblots containing recombinant P40 even 3 months after infection. The gene encoding P40 was present in a single copy in all of the 26 field strains of M. agalactiae analyzed and was not detected in closely related mycoplasma species. P40 was expressed as a protein with an apparent molecular mass of 37 kDa on sodium dodecyl sulfate-acrylamide gels by all M. agalactiae strains except for serotype C strains, which showed nonsense mutations in their p40 genes. PMID:12228289

  15. Genome Sequences of Staphylococcus hominis Strains ShAs1, ShAs2, and ShAs3, Isolated from the Asian Malaria Mosquito Anopheles stephensi

    PubMed Central

    Hughes, Grant L.; Raygoza Garay, Juan Antonio; Koundal, Vikas; Mwangi, Michael M.

    2016-01-01

    Staphylococcus hominis is a culturable component of the bacterial microbiome of Anopheles stephensi. Here, we present the annotated draft genome sequences of three S. hominis isolates from A. stephensi. These genomic resources will facilitate experiments to further our understanding of the role of bacteria in mosquito biology. PMID:26966197

  16. Genome Sequences of Staphylococcus hominis Strains ShAs1, ShAs2, and ShAs3, Isolated from the Asian Malaria Mosquito Anopheles stephensi.

    PubMed

    Hughes, Grant L; Raygoza Garay, Juan Antonio; Koundal, Vikas; Rasgon, Jason L; Mwangi, Michael M

    2016-03-10

    Staphylococcus hominis is a culturable component of the bacterial microbiome of Anopheles stephensi. Here, we present the annotated draft genome sequences of three S. hominis isolates from A. stephensi. These genomic resources will facilitate experiments to further our understanding of the role of bacteria in mosquito biology.

  17. Multilocus sequence typing of Mycoplasma hyorhinis strains identified by a real-time TaqMan PCR assay.

    PubMed

    Tocqueville, Véronique; Ferré, Séverine; Nguyen, Ngoc Hong Phuc; Kempf, Isabelle; Marois-Créhan, Corinne

    2014-05-01

    A real-time TaqMan PCR assay based on the gene encoding the protein p37 was developed to detect Mycoplasma hyorhinis. Its specificity was validated with 29 epidemiologically unrelated M. hyorhinis strains (28 field strains and one reference strain) and other mycoplasma species or with other microorganisms commonly found in pigs. The estimated detection limit of this qPCR assay was 125 microorganism equivalents/μl. The same 29 epidemiologically unrelated M. hyorhinis strains and four previously fully sequenced strains were typed by two portable typing methods, the sequencing of the p37 gene and a multilocus sequence typing (MLST) scheme. The first method revealed 18 distinct nucleotide sequences and insufficient discriminatory power (0.934). The MLST scheme was developed with the sequenced genomes of the M. hyorhinis strains HUB-1, GDL-1, MCLD, and SK76 and based on the genes dnaA, rpoB, gyrB, gltX, adk, and gmk. In total, 2,304 bp of sequence was analyzed for each strain. MLST was capable of subdividing the 33 strains into 29 distinct sequence types. The discriminatory power of the method was >0.95, which is the threshold value for interpreting typing results with confidence (D=0.989). Population analysis showed that recombination in M. hyorhinis occurs and that strains are diverse but with a certain clonality (one unique clonal complex was identified). The new qPCR assay and the robust MLST scheme are available for the acquisition of new knowledge on M. hyorhinis epidemiology. A web-accessible database has been set up for the M. hyorhinis MLST scheme at http://pubmlst.org/mhyorhinis/.

  18. 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

  19. Whole-genome sequence of Staphylococcus hominis, an opportunistic pathogen.

    PubMed

    Jiang, Saiping; Zheng, Beiwen; Ding, Wenchao; Lv, Longxian; Ji, Jinru; Zhang, Hua; Xiao, Yonghong; Li, Lanjuan

    2012-09-01

    Staphylococcus hominis is a commensal coagulase-negative species of staphylococci. It has been considered a presumptive and opportunistic pathogen that causes nosocomial infections in humans. Here we present the draft genome sequence of S. hominis ZBW5, a multidrug-resistant strain isolated from a human skin sample, which provides opportunities to understand the mechanism and genetic basis of its pathogenesis.

  20. 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

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

    PubMed

    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/μl) and a quantitative multivariate detection limit of 5.3 ± 1 cells/μl. 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

  2. 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/μl) and a quantitative multivariate detection limit of 5.3 ± 1 cells/μl. 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

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

    PubMed

    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/μl) and a quantitative multivariate detection limit of 5.3 ± 1 cells/μl. 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.

  4. Performance evaluation of two microbial transport media designed for preservation and transport of Chlamydiae, Mycoplasma and Ureaplasma.

    PubMed

    Jones, Sara L; Madhusudhan, Kunapuli T; Agans, Krystle; Dearen, Karen; Knight, Jennifer; Brasel, Trevor; Karamchi, Mehdi; Sherwood, Robert L

    2015-04-01

    The ability of a non-propagating transport device (test device) to maintain the viability of clinically relevant bacteria was compared with a similar commercial device (predicate device) to establish performance equivalence. Test bacteria, namely Chlamydia trachomatis, Chlamydia pneumoniae, Mycoplasma hominis, Mycoplasma pneumoniae and Ureaplasma urealyticum, were inoculated into the test [Puritan Medical Products Universal Transport System (UniTranz-RT(TM))] and predicate (BD Universal Viral Transport System) devices, and incubated at 4 °C and room temperature for up to 72 h. Bacterial viability was assessed at selected time points post-incubation using shell vial assays followed by immunofluorescence staining (for Chlamydia) or by standard culture techniques (for Mycoplasma and Ureaplasma). Results indicated that the Chlamydia strains were equally stable in both test and predicate devices through 72 h storage, at both test temperatures. Quantifiable levels of Mycoplasma and Ureaplasma were recovered from the test and predicate devices throughout the storage period. Low-temperature storage improved bacterial viability when compared with room temperature storage. In addition, the predicate device demonstrated slightly improved performance versus the test device in the context of Mycoplasma and Ureaplasma following 72 h storage. The overall results of the study confirmed the full performance of UniTranz-RT(TM) as a microbial transport medium and established equal performance with the predicate device.

  5. Short communication: In vitro antimicrobial susceptibility of Mycoplasma agalactiae strains isolated from dairy goats.

    PubMed

    Paterna, A; Sánchez, A; Gómez-Martín, A; Corrales, J C; De la Fe, C; Contreras, A; Amores, J

    2013-01-01

    This study examined the susceptibility to several antimicrobials of 28 isolates of Mycoplasma agalactiae obtained from goats in a region (southeastern Spain) where contagious agalactia is endemic. For each isolate, the minimum inhibitory concentration (MIC) against 12 antimicrobials of the quinolone, macrolide, aminoglycoside, and tetracycline families was determined. The antimicrobials with the lowest MIC were enrofloxacin, ciprofloxacin, tylosin, and doxycycline, all with MIC90 (concentration at which growth of 90% of the isolates is inhibited) <1 µg/mL. Norfloxacin (a quinolone) showed a wide MIC range (0.1-12.8 µg/mL), suggesting a resistance mechanism toward this antimicrobial that was not elicited by enrofloxacin or ciprofloxacin (the other quinolones tested). Erythromycin showed the highest MIC90 such that its use against Mycoplasma agalactiae is not recommended. Finally, Mycoplasma agalactiae isolates obtained from goat herds with clinical symptoms of contagious agalactia featured higher MIC90 and MIC50 (concentration at which growth of 50% of the isolates is inhibited) values for many of the antimicrobials compared with isolates from asymptomatic animals. The relationship between the extensive use of antimicrobials in herds with clinical contagious agalactia and variations in MIC requires further study. PMID:24035026

  6. [Dynamics of Mycoplasma infection in patients with glomerulo- nephritis during pathogenetic therapy].

    PubMed

    Pyrig, L A; Rudenko, A V; Nikonova, N A; Kudriavskaia, V M

    1989-11-01

    Among bacterial infections arising in glomerulonephritis and complicating its course Mycoplasma infection (M. hominis) is not a rare finding as shown by microbiological and serological examinations. Good therapeutic results achieved in patients treated with prednisolone and cyclophosphamide suggest inhibiting action of glucocorticoids on Mycoplasma infection.

  7. [Blastocystis hominis and bowel diseases].

    PubMed

    Ustün, Sebnem; Turgay, Nevin

    2006-01-01

    Blastocystis hominis (B. hominis) is a parasite of uncertain role in human disease. It may be identified during a workup for gastrointestinal symptoms, usually in stools. The clinical consequences of B. hominis infection are mainly diarrhea and abdominal pain as well as nonspecific gastrointestinal symptoms such as nausea, anorexia, vomiting, weight loss, lassitude, dizziness, and flatulence. Case reports and series have suggested a pathogenic role of B. hominis in causing intestinal inflammation. Also some studies have suggested that inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS) are associated with B. hominis infection. The investigators indicate that the stools of all patients presenting with IBD or IBS should be examined, and culture methods for B. hominis carried out. Invasion and mucosal inflammation of the intestine with B. hominis have been observed in studies of gnotobiotic guinea pigs. The transmission, pathogenicity, culture characteristics, taxonomy, life cycle, biochemistry and molecular biology of B. hominis remain unclear. More studies are necessary for this parasite. PMID:17106862

  8. Clonal Spread of a Unique Strain of Macrolide-Resistant Mycoplasma Pneumoniae Within a Single Family in Italy.

    PubMed

    Chironna, Maria; Loconsole, Daniela; De Robertis, Anna Lisa; Morea, Anna; Scalini, Egidio; Quarto, Michele; Tafuri, Silvio; Germinario, Cinzia; Manzionna, Mariano

    2016-03-01

    Macrolide-resistant Mycoplasma pneumoniae (MR-MP) is an increasing problem worldwide. This study describes the clonal spread of a unique strain of MR-MP within a single family. On January 23, 2015, nasopharyngeal swabs and sputum samples were collected from the index case (a 9-year-old girl) in southern Italy. The patient had pneumonia and was initially treated with clarithromycin. MR-MP infection was suspected due to prolonged symptoms despite appropriate antibiotic therapy. Two further cases of pneumonia occurred in relatives (a 7-year-old cousin and the 36-year-old mother of the index case); therefore, respiratory samples were also collected from other family members. Sequence analysis identified mutations associated with resistance to macrolides. Both P1 major adhesion protein typing and multiple loci variable-number tandem repeat analysis (MLVA) typing were performed to assess the relatedness of the strains. The index case, the cousin, the mother, and another 4 family members (twin siblings of the index case, a 3-year-old cousin, and the grandmother) were positive for MR-MP. All strains harbored the mutation A2063G, had the same P1 subtype (1), and were MLVA (7/4/5/7/2) type Z. In addition, the index case's aunt (31 years of age and the probable source of infection) harbored an M pneumoniae strain with the same molecular profile; however, this strain was susceptible to macrolides. This cluster of MR-MP infection/carriage caused by a clonal strain suggests a high transmission rate within this family and highlights the need for increased awareness among clinicians regarding the circulation of MR-MP. Novel strategies for the treatment and prevention of M pneumoniae infections are required. PMID:26986172

  9. Clonal Spread of a Unique Strain of Macrolide-Resistant Mycoplasma Pneumoniae Within a Single Family in Italy

    PubMed Central

    Chironna, Maria; Loconsole, Daniela; De Robertis, Anna Lisa; Morea, Anna; Scalini, Egidio; Quarto, Michele; Tafuri, Silvio; Germinario, Cinzia; Manzionna, Mariano

    2016-01-01

    Abstract Macrolide-resistant Mycoplasma pneumoniae (MR-MP) is an increasing problem worldwide. This study describes the clonal spread of a unique strain of MR-MP within a single family. On January 23, 2015, nasopharyngeal swabs and sputum samples were collected from the index case (a 9-year-old girl) in southern Italy. The patient had pneumonia and was initially treated with clarithromycin. MR-MP infection was suspected due to prolonged symptoms despite appropriate antibiotic therapy. Two further cases of pneumonia occurred in relatives (a 7-year-old cousin and the 36-year-old mother of the index case); therefore, respiratory samples were also collected from other family members. Sequence analysis identified mutations associated with resistance to macrolides. Both P1 major adhesion protein typing and multiple loci variable-number tandem repeat analysis (MLVA) typing were performed to assess the relatedness of the strains. The index case, the cousin, the mother, and another 4 family members (twin siblings of the index case, a 3-year-old cousin, and the grandmother) were positive for MR-MP. All strains harbored the mutation A2063G, had the same P1 subtype (1), and were MLVA (7/4/5/7/2) type Z. In addition, the index case's aunt (31 years of age and the probable source of infection) harbored an M pneumoniae strain with the same molecular profile; however, this strain was susceptible to macrolides. This cluster of MR-MP infection/carriage caused by a clonal strain suggests a high transmission rate within this family and highlights the need for increased awareness among clinicians regarding the circulation of MR-MP. Novel strategies for the treatment and prevention of M pneumoniae infections are required. PMID:26986172

  10. Susceptibility of alpine ibex to conjunctivitis caused by inoculation of a sheep-strain of Mycoplasma conjunctivae.

    PubMed

    Giacometti, M; Nicolet, J; Frey, J; Krawinkler, M; Meier, W; Welle, M; Johansson, K E; Degiorgis, M P

    1998-04-15

    We evaluated the susceptibility of alpine ibex (Capra ibex ibex) to mycoplasmal conjunctivitis induced by a strain of Mycoplasma conjunctivae isolated from domestic sheep by inoculation of three alpine ibexes with 1.2 x 10(6) colony forming units of M. conjunctivae in the conjunctival sac of both eyes. One more ibex was exposed to the infection by contact. Experimental animals were free of M. conjunctivae and ocular Chlamydia infection before inoculation. Conjunctivitis and serous to mucous lachrymation became apparent in all four ibexes. Clinical signs began within 2 days in inoculated animals and 22 days after the beginning of the experiment in the contact ibex. M. conjunctivae was demonstrated up to the 63th day post-inoculation by cultural and PCR-methods. After 63 days, histopathologic examination revealed nearly normal ocular tissues, and M. conjunctivae could be detected from two eyes only. No other infectious agents which might cause conjunctivitis or keratitis, including Chlamydia psittaci and Branhamella ovis, were involved. Our investigation indicates that sheep-strains of M. conjunctivae can induce conjunctivitis in alpine ibex, thus showing pathogenicity of this organism for Caprinae species other than domestic sheep and goats.

  11. Staphylococcus hominis subsp. novobiosepticus subsp. nov., a novel trehalose- and N-acetyl-D-glucosamine-negative, novobiocin- and multiple-antibiotic-resistant subspecies isolated from human blood cultures.

    PubMed

    Kloos, W E; George, C G; Olgiate, J S; Van Pelt, L; McKinnon, M L; Zimmer, B L; Muller, E; Weinstein, M P; Mirrett, S

    1998-07-01

    A new subspecies, Staphylococcus hominis subsp. novobiosepticus, isolated from human blood cultures, a wound, a breast abscess and a catheter tip, is described on the basis of a study of 26 strains isolated between 1989 and 1996. DNA-DNA reassociation reactions, conducted under stringent conditions, and macrorestriction pattern analysis demonstrated that these strains are closely related to previously characterized S. hominis strains isolated from human skin and clinical specimens, but are significantly divergent. S. hominis subsp. novobiosepticus can be distinguished from S. hominis (now named S. hominis subsp. hominis) by its combined characteristics of novobiocin resistance and failure to produce acid aerobically from D-trehalose and N-acetyl-D-glucosamine. Furthermore, all 26 strains of the new subspecies are resistant to nalidixic acid, penicillin G, oxacillin, kanamycin and streptomycin, and were either resistant or had intermediate resistance to methicillin and gentamicin. Most strains were also resistant to erythromycin, clindamycin, chloramphenicol, trimethoprim/sulfamethoxazole and ciprofloxacin. Based on a comparison of the sequences of a 1001 bp mecA amplification product from reference methicillin-resistant staphylococci, the mecA gene present in S. hominis subsp. novobiosepticus was identified as homologue A, commonly found in S. aureus and many coagulase-negative staphylococcal species. The type strain of S. hominis subsp. novobiosepticus is ATCC 700236T. Descriptions of S. hominis subsp. novobiosepticus subsp. nov and S. hominis subsp. hominis are given and the description of S. hominis is emended.

  12. Strain differences in sensitivity of rats to Mycoplasma arthritidis ISR 1 infection are under multiple gene control.

    PubMed Central

    Binder, A; Gärtner, K; Hedrich, H J; Hermanns, W; Kirchhoff, H; Wonigeit, K

    1990-01-01

    At least 5 female rats from each of 24 inbred (ACI, AS, BDIX, BH, BN, BS, BUF, DA, LE, LEW, MWF, OM, SPRD-Cu3, W-Krypt, and WKY), RT1 congenic [BH.1L(LEW), LEW.1A(AVN), LEW.1C(WIST), LEW.1LV3(BH), LEW.1K(SHR), and LEW.1N(BN)], and F1 hybrid [(LEW x BN)F1, (LEW.1W x LEW.1A)F1, and (LEW x LEW.1W)F1] strains, representing eight independent major histocompatibility complex (MHC) haplotypes (a, b, c, dv1, k, l, n, and u) and five related RT1 haplotypes (av1, lv1, lv3, uv2, and uv3), were inoculated intravenously with Mycoplasma arthritidis, and the severity of the polyarthritis that developed was determined by estimating arthritis scores and weight reductions. The 24 inbred, congenic, and F1 hybrid rat strains differed considerably in their sensitivity to infection with M. arthritidis and in the severity of the polyarthritis that they developed. Statistical evaluation showed that in the acute phase (days 1 to 42 after infection) as well as in the chronic phase (days 39 to 121 after infection) of the disease, the means of the arthritis scores for the strains form a continuous variation without significant interruptions, with the very sensitive LEW rats, the RT1 congenic rats on LEW background, the F1 hybrids with LEW, and the MWF, BS, BH, and DA rats on one end and the resistant WKY, BUF, W-Krypt, LE, and OM rats on the other end. A continuous variation was also observed for the means of the growth rates. There were, however, no significant differences between the sensitive and the resistant rat strains in the antibody titers determined by complement fixation test and enzyme immunoassay. Heritabilities of arthritis scores were calculated for all strains (h2 = 0.39 to 0.62), for the RT1 congenic strains (h2 = 0.04 to 0.14), and for several strains with identical MHC genes (h2 = 0.61 to 0.93). The results show that non-MHC genes are probably responsible for the sensitivity of rats to infection with M. arthritidis. PMID:2111282

  13. Susceptibility of goats and calves after experimental inoculation or contact exposure to a Canadian strain of Mycoplasma mycoides subsp. mycoides isolated from a goat.

    PubMed Central

    Rosendal, S

    1983-01-01

    Transmissibility of Mycoplasma mycoides subsp. mycoides infection from experimentally inoculated goats to other goats and calves was studied. Eight goats and six calves were housed in an 18 m2 room. Six of the goats were inoculated endobronchially with strain D44 isolated from a natural case of polyarthritis in Ontario. These six goats died within a week of Mycoplasma septicemia. The two contact goats or the six calves never showed signs of disease and M. mycoides subsp. mycoides was not recovered from these animals. The contact goats and four calves were killed 25 days after exposure. They were all seronegative, M. mycoides subsp. mycoides was not recovered at necropsy and none had pathomorphological changes attributable to this Mycoplasma. The two remaining calves were inoculated endobronchially with 10(9) CFU of strain D44 and observed for 20 days. They never showed signs of disease and did not have significant lesions at necropsy. Both developed a significant serological response to M. mycoides subsp. mycoides, although this organism was not recovered during the experimental period or at necropsy. This study did not provide evidence for transmission of M. mycoides subsp. mycoides from endobronchially inoculated goats to contact goats or calves and endobronchially inoculated calves did not develop pneumonia. This would suggest that the infection of the goat population in Canada with this pathogen would not be a significant threat to the cattle population. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. Fig. 7. PMID:6365296

  14. Blastocystis hominis revisited.

    PubMed Central

    Stenzel, D J; Boreham, P F

    1996-01-01

    Blastocystis hominis is a unicellular organism found commonly in the intestinal tract of humans and many other animals. Very little is known of the basic biology of the organism, and controversy surrounds its taxonomy and pathogenicity. There morphological forms (vacuolar, granular, and ameboid) have been recognized, but recent studies have revealed several additional forms (cyst, avacuolar, and multivacuolar). The biochemistry of the organism has not been studied to any extent, and organelles and structures of unknown function and composition are present in the cells. Several life cycles have been proposed but not experimentally validated. The form used for transmission has not been defined. Infections with the organism are worldwide and appear in both immunocompetent and immunodeficient individuals. Symptoms generally attributed to B. hominis infection are nonspecific, and the need for treatment is debated. If treatment appears warranted, metronidazole is suggested as the drug of choice, although failures of this drug in eradicating the organism have been reported. Infection is diagnosed by light microscopic examination of stained smears or wet mounts of fecal material. Most laboratories identify B. hominis by observing the vacuolar form, although morphological studies indicate that other forms, such as the cyst form and multivacuolar form, also should be sought for diagnosis. PMID:8894352

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  16. Draft Genome Sequence of the Multidrug-Resistant Clinical Isolate Dermabacter hominis 1368

    PubMed Central

    Albersmeier, Andreas; Bomholt, Christina; Glaub, Alina; Rückert, Christian; Soriano, Francisco; Fernández-Natal, Isabel

    2014-01-01

    Dermabacter hominis is a common colonizer of the healthy human skin and is rarely detected as an opportunistic human pathogen. The genome sequence of the multidrug-resistant D. hominis strain 1368, isolated from blood cultures of a pyelonephritis patient, provides insights into the repertoire of antibiotic resistance genes. PMID:25059872

  17. A high level of strain variation within the Mycoplasma ovipneumoniae population of the UK has implications for disease diagnosis and management.

    PubMed

    Parham, K; Churchward, Colin P; McAuliffe, L; Nicholas, R A J; Ayling, R D

    2006-11-26

    Mycoplasma ovipneumoniae is one of only two mycoplasma species associated with small ruminant disease in Britain and has been associated with an increasing number of disease outbreaks since 2002. This investigation used well-defined techniques to assess the variability of UK M. ovipneumoniae isolates, in an attempt to identify strain clusters within the population. Strains received for routine diagnosis between 2002 and 2004 were analysed using random amplified polymorphic DNA (RAPD) and pulsed field gel electrophoresis (PFGE). Of the 43 samples screened 40 RAPD Hum-1, 41 RAPD Hum-4 and 40 PFGE profiles were observed. Composite data analysis divided strains into 10 similarity clusters with SDS-PAGE and Western blotting indicating that this DNA variability is translated into a pattern of variable protein expression. In order to assess the strains isolated within flocks two sets of samples, from diverse locations, were included in this test panel. The presence of variable isolates existing on the same farm may reflect animal movement and the introduction of asymptomatic, carrier, animals where M. ovipneumoniae is already established within a flock. These findings have significant implications regarding disease diagnosis and management.

  18. Comparison of the new Mycofast Revolution assay with a molecular assay for the detection of genital mycoplasmas from clinical specimens

    PubMed Central

    2013-01-01

    Background Genital mycoplasmas are opportunistic bacteria that are associated with undesirable gynaecologic and reproductive events. Mycoplasmas are fastidious bacteria with increasing resistance to routine antimicrobials and often fail to grow on conventional culture methods. The commercial Mycofast Revolution assay permits the phenotypic detection and identification of genital mycoplasmas. Antimicrobial susceptibility testing against five antimicrobial agents with MICs corresponding to the CLSI guidelines can also be performed. This study aimed to compare the new commercially available Mycofast Revolution assay with a multiplex PCR assay. Methods Self-collected swabs were obtained from pregnant women attending the antenatal clinic of a tertiary academic hospital in Pretoria, South Africa from October 2012 to November 2012. These swabs were used to seed UMMt and modified Amies transport media. The seeded UMMt transported medium was used to inoculate the Mycofast Revolution assay for the identification, enumeration and antimicrobial susceptibility testing of genital mycoplasmas. Following DNA extraction from the modified Amies transport medium, specimens were subjected to a multiplex PCR assay for the detection of genital mycoplasmas. Results The Mycofast Revolution kit had a sensitivity and specificity of 77.3% (95% CI: 62.15% to 88.51%) and 80% (95% CI: 28.81% to 96.70%), respectively, against the PCR assay. The positive and negative predictive values were 97.1% (95% CI: 85.03% to 99.52%) and 28.6% (95% CI: 8.57% to 58.08%). Genital mycoplasmas were detected in 71.4% (35/49) of samples with the Mycofast Revolution assay with 49% (24/49) being Ureaplasma spp. and 22.4% (11/49) mixed strains. The multiplex PCR assay had a positivity rate of 89.8% (44/49) for genital mycoplasmas; mixed strains were present in 51% (25/49) of samples, Ureaplasma spp. in 16.3% (8/49) and M. hominis in 22.4% (11/49) of samples. Conclusions There was a fair agreement (κ = 0

  19. Polypeptide and antigenic variability among strains of Mycoplasma ovipneumoniae demonstrated by SDS-PAGE and immunoblotting.

    PubMed

    Thirkell, D; Spooner, R K; Jones, G E; Russell, W C

    1990-01-01

    Comparison of the polypeptide patterns of 22 isolates of M. ovipneumoniae by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a marked degree of heterogeneity with only limited groupings identifiable. Of the 50 major polypeptides identified in one strain (956/2), 35 were shown to be antigenic using immunoblotting with a homologous polyclonal serum. Radioimmune precipitation of 125I-surface-labelled proteins and phase partition using Triton X-114 detergent indicated that these were membrane associated. Cross-reactivity between the isolates was examined by immunoblotting using one polyclonal serum and four monoclonal antibodies (MAbs), all raised against strain 956/2. The polyclonal serum revealed considerable antigenic heterogeneity, but at least nine major antigens were conserved across all isolates. Two MAbs cross-reacted with all 22 strains, but the other two MAbs allowed some differentiation of the strains. One (MO/3) divided the isolates into groups of 16 and 6 based on the presence of absence of a 26-kDa antigen. All strains isolated from sheep with pulmonary adenomatosis fell into the smaller group and did not possess the 26-kDa antigen.

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

    PubMed

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

    2016-02-01

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

  1. Prevalence of genital mycoplasmas, ureaplasmas and chlamydia in pregnancy.

    PubMed

    Govender, S; Theron, G B; Odendaal, H J; Chalkley, L J

    2009-11-01

    The study was designed to determine the prevalence of genital mycoplasmas, ureaplasmas and Chlamydia on women attending their first prenatal visit, in conjunction with pre-term labour or HIV status. For pre-term labour (2003), 199 women were monitored for pre-term delivery (<37 weeks); for colonisation and HIV (2005), 219 women were screened. Microbial detection was performed on DNA extracted from endocervical swabs employing PCR techniques. Colonisation was seen to be highest in the 14-20 year age group from 2003. In women aged > or = 21 years, co-colonisation was 13%, although there was a shift from co-colonisation with Mycoplasma hominis and Ureaplasma urealyticum in 2003, to other dual/triple combinations in 2005. Overall, major trends from both collection periods were that the prevalence of U. urealyticum tended to be higher in women > or = 26 years, while the prevalence of Chlamydia trachomatis and M. hominis lower. No association was evident between colonisation with M. hominis, U. urealyticum, Ureaplasma parvum and labour outcome. HIV status had no effect on the prevalence/co-colonisation of M. hominis, U. urealyticum or C. trachomatis. The importance of genital mycoplasmas, ureaplasmas and C. trachomatis in long-term aetiologies requires further investigations, certainly in relation to syndromic management regimens that fail to reduce colonisation rates. PMID:19821660

  2. Rapid, Simple and Cost-Effective Molecular Method to Differentiate the Temperature Sensitive (ts+) MS-H Vaccine Strain and Wild-Type Mycoplasma synoviae Isolates.

    PubMed

    Kreizinger, Zsuzsa; Sulyok, Kinga Mária; Pásztor, Alexandra; Erdélyi, Károly; Felde, Orsolya; Povazsán, János; Kőrösi, László; Gyuranecz, Miklós

    2015-01-01

    Mycoplasma synoviae infection in chickens and turkeys can cause respiratory disease, infectious synovitis and eggshell apex abnormality; thus it is an economically important pathogen. Control of M. synoviae infection comprises eradication, medication or vaccination. The differentiation of the temperature sensitive (ts+) MS-H vaccine strain from field isolates is crucial during vaccination programs. Melt-curve and agarose gel based mismatch amplification mutation assays (MAMA) are provided in the present study to distinguish between the ts+ MS-H vaccine strain, its non-temperature sensitive re-isolates and wild-type M. synoviae isolates based on the single nucleotide polymorphisms at nt367 and nt629 of the obg gene. The two melt-MAMAs and the two agarose-MAMAs clearly distinguish the ts+ MS-H vaccine strain genotype from its non-temperature sensitive re-isolate genotype and wild-type M. synoviae isolate genotype, and no cross-reactions with other Mycoplasma species infecting birds occur. The sensitivity of the melt-MAMAs and agarose-MAMAs was 103 and 104 copy numbers, respectively. The assays can be performed directly on clinical samples and they can be run simultaneously at the same annealing temperature. The assays can be performed in laboratories with limited facilities, using basic real-time PCR machine or conventional thermocycler coupled with agarose gel electrophoresis. The advantages of the described assays compared with previously used methods are simplicity, sufficient sensitivity, time and cost effectiveness and specificity. PMID:26207635

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

    PubMed

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

    2011-01-01

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

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

    PubMed Central

    2011-01-01

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

  5. Mycoplasma and ureaplasma infection and male infertility: a systematic review and meta-analysis.

    PubMed

    Huang, C; Zhu, H L; Xu, K R; Wang, S Y; Fan, L Q; Zhu, W B

    2015-09-01

    The relationship between mycoplasma and ureaplasma infection and male infertility has been studied widely; however, results remain controversial. This meta-analysis investigated the association between genital ureaplasmas (Ureaplasma urealyticum, Ureaplasma parvum) and mycoplasmas (Mycoplasma hominis, Mycoplasma genitalium), and risk of male infertility. Differences in prevalence of ureaplasma and mycoplasma infection between China and the rest of the world were also compared. Study data were collected from PubMed, Embase and the China National Knowledge Infrastructure. Summary odds ratio (OR) with 95% confidence interval (CI) was applied to assess the relationship. Heterogeneity testing and publication bias testing were also performed. A total of 14 studies were used: five case-control studies with 611 infertile cases and 506 controls featuring U. urealyticum infection, and nine case-control studies with 2410 cases and 1223 controls concerning M. hominis infection. Two other infection (U. parvum and M. genitalium) were featured in five and three studies, respectively. The meta-analysis results indicated that U. parvum and M. genitalium are not associated with male infertility. However, a significant relationship existed between U. urealyticum and M. hominis and male infertility. Comparing the global average with China, a significantly higher positive rate of U. urealyticum, but a significantly lower positive rate of M. hominis, was observed in both the infertile and control groups in China.

  6. 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...

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

    PubMed Central

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

    2016-01-01

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

  8. Previously Uncharacterized Mycoplasma Isolates from an Investigation of Canine Pneumonia

    PubMed Central

    Armstrong, D.; Tully, J. G.; Yu, B.; Morton, V.; Friedman, M. H.; Steger, L.

    1970-01-01

    Mycoplasmas recovered from the respiratory tract and genitourinary system of dogs, with and without respiratory infection, have been characterized by biological and immunological methods. Some of the isolates were indentified as being similar to the three species of canine mycoplasmas described earlier under the designation Mycoplasma spumans, M. canis, and M. maculosum. Other mycoplasmas placed in three groups (A, C, and D) were found to be clearly distinct from the three classified species. Group A strains fermented glucose but not mannose and were serologically distinct from other canine mycoplasmas recovered in this study. These strains were subsequently found to be biologically and serologically related to a previously reported, but unclassified, canine mycoplasma. Group D strains differed in some biological properties but were serologically related. These were found to be nonfermenting mycoplasmas representing isolations from the throat and bladder of dogs. They were serologically distinct from other canine mycoplasmas and were apparently unrelated to other known mycoplasma serotypes. Group C mycoplasmas were recovered only from the lungs of dogs. Within the group, they differ in some immunological properties but appear to be serologically distinct from other canine strains. They can also be separated from other dog strains in their ability to ferment glucose and mannose. Group B strains were found to have biological properties similar to M. canis strains but seemed to be only partially related to this serotype when examined in several serological techniques. It is suggested that these strains might represent antigenic variants of M. canis. PMID:16557682

  9. Blastocystis hominis in hospital employees.

    PubMed

    Grossman, I; Weiss, L M; Simon, D; Tanowitz, H B; Wittner, M

    1992-06-01

    Several reports have appeared that either support or deny the importance of the protozoan Blastocystis hominis as an intestinal pathogen in humans. In this report, we describe the clinical characteristics of B. hominis and its response to therapy in hospital employees found to have the parasite on routine screening of stools. During the study, 49 patients with B. hominis were identified, and 413 stools were examined from these patients. Twenty-nine patients were asymptomatic (59%), and 20 had symptoms of bloating, flatulence, soft/loose stools, or constipation. Of these 20 patients, 10 had symptoms that correlated with the presence or absence of B. hominis, four had symptoms that were independent of B. homonis, and six had other intestinal parasites that could account for their symptoms. Nineteen percent of patients without treatment had eradication of B. hominis from stool on follow-up examination. Metronidazole did not increase this rate. Iodoquinol treatment eradicated the organism in 41% of patients (p less than 0.05), and resulted in the reduction or eradication of the parasite in 62%, as determined by follow-up examination. PMID:1590309

  10. Two strains of Mycoplasma synoviae from chicken flocks on the same layer farm differ in their ability to produce eggshell apex abnormality.

    PubMed

    Catania, S; Gobbo, F; Bilato, D; Gagliazzo, L; Moronato, M L; Terregino, C; Bradbury, J M; Ramírez, A S

    2016-09-25

    Mycoplasma synoviae (Ms) is considered to be an economically important poultry pathogen. Although the full economic costs of infection in layer chickens are still under debate, the prevalence of Ms is known to be high in some countries and earlier reports have shown a correlation between infection and Eggshell Apex Abnormality (EAA). This work is a continuation of an earlier study of a clinical case of EAA on a layer hen farm where the presence of two different strains of Ms, based on the sequence of the 5' end of the vlhA gene, was demonstrated. Both strains could be detected in the trachea but only one (designated strain PASC8) appeared able to colonize the oviduct, while the other (designated TRACH) was not found in the oviduct and has not been related to EAA. The PASC8 partial vlhA gene sequence differs from that of the TRACH in having a 39 nucleotide deletion in the proline rich region and three point mutations in the RIII region. Based on this information an experimental infection was performed in SPF chickens using groups infected with either the PASC8 or the TRACH strain and a non-infected control group. Both Ms strains were detected in the trachea of infected birds, but only the PASC8 strain was found in the oviduct. Furthermore, EAA developed only in the group infected with PASC8 strain. Compared to the control group, both strains produced an adverse impact on egg production: a decrease in the numbers laid and in their average weight (P<0.05) This work demonstrates a difference in oviduct tropism between two Ms strains and a possible relationship to the production of EAA in experimental conditions. PMID:27599931

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed

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

    2014-01-01

    Mycoplasma spp. are highly contagious pathogens and intramammary Mycoplasma infection is a serious issue for the dairy industry. As there is no effective vaccine for Mycoplasma infection, control depends on good husbandry and chemo-antibiotic therapy. In this study, antimicrobial susceptibility of Mycoplasma strains recently isolated from cases of bovine mastitis in Japan was evaluated by minimum inhibitory concentration (MIC). All Mycoplasma bovis strains were sensitive to pirlimycin, danofloxacin and enrofloxacin, but not kanamycin, oxytetracycline, tilmicosin or tylosin. M. californicum and M. bovigenitalium strains were sensitive to pirlimycin, danofloxacin, enrofloxacin, oxytetracycline, tilmicosin and tylosin, but not to kanamycin. This is the first report to describe the MIC of major antimicrobial agents for Mycoplasma species isolated from bovine mastitis in Japan. PMID:24261609

  13. Urogenital Mycoplasmas and Human Papilloma Virus in Hemodialysed Women

    PubMed Central

    Ekiel, Alicja; Pietrzak, Bronisława; Aptekorz, Małgorzata; Mazanowska, Natalia; Kamiński, Paweł; 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

  14. Survey of Surface Proteins from the Pathogenic Mycoplasma hyopneumoniae Strain 7448 Using a Biotin Cell Surface Labeling Approach

    PubMed Central

    Reolon, Luciano Antonio; Martello, Carolina Lumertz; Schrank, Irene Silveira; Ferreira, Henrique Bunselmeyer

    2014-01-01

    The characterization of the repertoire of proteins exposed on the cell surface by Mycoplasma hyopneumoniae (M. hyopneumoniae), the etiological agent of enzootic pneumonia in pigs, is critical to understand physiological processes associated with bacterial infection capacity, survival and pathogenesis. Previous in silico studies predicted that about a third of the genes in the M. hyopneumoniae genome code for surface proteins, but so far, just a few of them have experimental confirmation of their expression and surface localization. In this work, M. hyopneumoniae surface proteins were labeled in intact cells with biotin, and affinity-captured biotin-labeled proteins were identified by a gel-based liquid chromatography-tandem mass spectrometry approach. A total of 20 gel slices were separately analyzed by mass spectrometry, resulting in 165 protein identifications corresponding to 59 different protein species. The identified surface exposed proteins better defined the set of M. hyopneumoniae proteins exposed to the host and added confidence to in silico predictions. Several proteins potentially related to pathogenesis, were identified, including known adhesins and also hypothetical proteins with adhesin-like topologies, consisting of a transmembrane helix and a large tail exposed at the cell surface. The results provided a better picture of the M. hyopneumoniae cell surface that will help in the understanding of processes important for bacterial pathogenesis. Considering the experimental demonstration of surface exposure, adhesion-like topology predictions and absence of orthologs in the closely related, non-pathogenic species Mycoplasma flocculare, several proteins could be proposed as potential targets for the development of drugs, vaccines and/or immunodiagnostic tests for enzootic pneumonia. PMID:25386928

  15. 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

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

    PubMed

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

    2013-12-01

    Five Mycoplasma strains from wild Caprinae were analyzed: four from Alpine ibex (Capra ibex) which died at the Berlin Zoo between 1993 and 1994, one from a Rocky Mountain goat collected in the USA prior to 1987. These five strains represented a population different from the populations belonging to the 'Mycoplasma mycoides cluster' as tested using multi locus sequence typing, Matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis and DNA-DNA hybridization. Analysis of the 16S rRNA gene (rrs), genomic sequence based in silico as well as laboratory DNA-DNA hybridization, and the analysis of phenotypic traits in particular their exceptionally rapid growth all confirmed that they do not belong to any Mycoplasma species described to date. We therefore suggest these strains represent a novel species, for which we propose the name Mycoplasma feriruminatoris sp. nov. The type strain is G5847(T) (=DSM 26019(T)=NCTC 13622(T)) [corrected]. PMID:24016869

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

    PubMed

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

    2013-12-01

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

  18. Monoclonal antibodies to surface-exposes proteins of Mycoplasma mycoides subsp. mycoides (small-colony strain), which causes contagious bovine pleuropneumonia.

    PubMed Central

    Kiarie, M N; Rurangirwa, F R; Perryman, L E; Jasmer, D P; McGuire, T C

    1996-01-01

    Outbreaks of bovine pleuropneumonia caused by small-colony strains of Mycoplasma mycoides subsp. mycoides occur in Africa, and vaccination is used for control. Since protein subunits are needed to improve multivalent vaccines, monoclonal antibodies (MAbs) were made to facilitate protein identification and isolation. Eleven immunoglobulin M MAbs derived from mouse spleen donors immunized with disrupted whole organisms bound periodate-sensitive epitopes on externally exposed polysaccharide. Seven of these MAbs caused in vitro growth inhibition of M. mycoides subsp. mycoides; however, reaction with carbohydrate epitopes prevented their use in identifying proteins. Ten additional MAbs from mouse spleen donors immunized with Triton X-114-phase integral membrane proteins reacted with periodate-insensitive, proteinase K-sensitive epitopes. These MAbs were classified into three groups based on immunoblots of Triton X-114-phase proteins. One group reacted with 96-, 16-, and 15-kDa proteins. Another group reacted with 26-, 21-, and 16-kDa proteins, while a third group reacted only with 26- and 21-kDa proteins. One MAb from each group reacted with trypsinsensitive epitopes on live organisms, yet none caused in vitro growth inhibition. Representative MAbs reacted with all small-colony strains in immunoblots and did not react with large colony strains. However, these MAbs were not specific for small-colony strains, as proteins from two other M. mycoides cluster organisms were identified. Nevertheless, MAbs to surface-exposed epitopes on integral membrane proteins will be useful for isolation of these proteins for immunization, since one or more might induce growth-inhibiting antibodies or other protective responses. PMID:8914769

  19. Isolation and molecular identification of mycoplasma genitalium from the secretion of genital tract in infertile male and female

    PubMed Central

    Mohseni Moghadam, Naeime; Kheirkhah, Babak; Mirshekari, Toraj Reza; Fasihi Harandi, Majid; Tafsiri, Elham

    2014-01-01

    Background: Mycoplasmas can cause acute and chronic diseases at multiple sites with wide-range complications and have been implicated as cofactors in diseases. The infections influenced form genital mycoplasmas specifically Mycoplasma hominis and Mycoplasma genitalium potentially affect reproductive disorders, and infertility. Objective: Isolation and molecular identification of Mycoplasma genitalium from the genital tract of infertile male and vaginal discharge of infertile female referred to Infertility Center of Kerman in 2013. Materials and Methods: This study was a randomized, prospective study. We included 100 infertile male and 100 infertile female that were referred to the Infertility Center of Kerman. Then for isolation and molecular identification of Mycoplasma genitalium from urethral and vaginal discharge polymerase chain reaction was performed on Mycoplasma genus and genitalium. Results: From a total of 100 semen samples 45 patients (45%) were mycoplasma-positive and 13 (28.8%) were genitalium species positive. Also, from a total of 100 women samples 43 women (43%) were mycoplasma-positive and 10 (23.2%) were genitalium species positive. Positive samples were sequenced and phylogenetic tree was drawn. Conclusion: According to the results of this study, a high percentage of infertile male and female were infected with the Mycoplasma genitalium. For prevention of harmful and significant consequences of this infection, we suggest a screening program in symptomatic infertile couples. PMID:25469132

  20. Molecular cloning and characterization of a surface-localized adhesion protein in Mycoplasma bovis Hubei-1 strain.

    PubMed

    Zou, Xiaohui; Li, Yuan; Wang, Yang; Zhou, Yumei; Liu, Yang; Xin, Jiuqing

    2013-01-01

    Mycoplasma bovis (M. bovis) is an important pathogen that causes various bovine diseases, such as mastitis in cows and pneumonia in calves. The surface proteins are generally thought to play a central role in the pathogenesis of this organism. We screened the entire genome of M. bovis Hubei-1 and discovered a gene named vpmaX that encodes the 25 kDa variable surface lipoprotein A (VpmaX). Sequence analysis revealed that VpmaX contains several repetitive units and a typical bacterial lipoprotein signal sequence. The vpmaX gene was cloned and expressed in E. coli to obtain recombinant VpmaX (rVpmaX). Western blot analysis using a rabbit antibody against rVpmaX demonstrated that VpmaX is a membrane protein. Immunostaining visualized via confocal laser scanning microscopy showed that rVpmaX was able to adhere to embryonic bovine lung cells (EBL), and this was also confirmed by a sandwich ELISA. In summary, a surface-localized adhesion protein was identified in M. bovis Hubei-1.

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

    PubMed

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

    2013-12-27

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

  2. Mycoplasma genitalium: from Chrysalis to Multicolored Butterfly

    PubMed Central

    Taylor-Robinson, David; Jensen, Jørgen Skov

    2011-01-01

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

  3. 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...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  5. Effects of F-strain Mycoplasma gallisepticum inoculation at twelve weeks of age on egg yolk composition in commercial egg laying hens.

    PubMed

    Burnham, M R; Peebles, E D; Branton, S L; Maurice, D V; Gerard, P D

    2003-04-01

    In two trials, the effects of F-strain Mycoplasma gallisepticum (FMG) on the contents of egg yolks from commercial Single Comb White Leghorn laying hens were investigated over a production cycle. Ten hens were assigned to each of 8 (trial 1) or 16 (trial 2) negative pressure fiberglass biological isolation units. Birds in half of the total units served as sham-inoculated controls, and those in the other half were inoculated with FMG at 12 wk of age. Eggs were collected and yolks were harvested at various times during the prepeak, peak, and postpeak periods of both trials for constituent analysis. Yolk constituents analyzed in these trials included moisture, total lipids, cholesterol, triglycerides, phospholipids, and fatty acids. In both trials, total yolk lipid at 22 wk of age was significantly decreased in birds inoculated with FMG. In trial 1, yolk cholesterol at 28 wk was significantly decreased in FMG-inoculated birds. Yolk linoleic acid in trial 1 and yolk stearic and arachidonic acids in trial 2 were significantly increased in FMG-inoculated birds compared to FMG-free birds. In trial 2, yolk myristic, palmitoleic, and oleic acid percentages were significantly decreased in FMG-inoculated birds compared to FMG-free birds. These data suggest that alterations in egg production in commercial layers in response to an FMG infection at 12 wk of age are associated with changes in yolk composition. PMID:12710476

  6. Emergence, re-emergence, spread and host species crossing of Mycoplasma bovis in the Austrian Alps caused by a single endemic strain.

    PubMed

    Spergser, Joachim; Macher, Kathrin; Kargl, Munkhtsetseg; Lysnyansky, Inna; Rosengarten, Renate

    2013-06-28

    Mycoplasma (M.) bovis was identified and reported in Austria as agent of infection and disease in cattle only once, namely in 2005 associated with a case of mastitis in a smallholding, but in 2007 it unexpectedly emerged as the cause of a devastating disease outbreak in a dairy herd of 100 individuals and spill over infection to pigs, both kept on the same mountain pasture. In 2008, M. bovis remained endemic at a low level in this region followed by the re-emergence of the agent in 2009 and a dramatic spread of the disease to further Alpine areas and their foothills in 2010 and 2011. From these outbreaks, a total of 94 M. bovis isolates including 7 porcine isolates were selected for genotyping. Two molecular tools, randomly amplified polymorphic DNA (RAPD) analysis and multi-locus variable number of tandem-repeat analysis (MLVA) were chosen to identify strain types involved in these outbreaks and to trace routes of infection and dynamics of dissemination. With both typing methods, the majority of Alpine isolates (96.8%) recovered over time from different areas and hosts was clustered into one group exhibiting a unique and indistinguishable profile which significantly differed from those of geographically unrelated strains including the type strain PG45 and 3 Alpine isolates which suddenly appeared and disappeared in 2009. Stability of the unique profile strongly indicated that a single M. bovis strain initiated the outbreak in 2007, crossed the host species barrier by infecting pigs, re-emerged in 2009 and became widespread in the Austrian Alps in 2010 and 2011. The remarkable dissemination and persistence of a single and unique M. bovis strain may reflect peculiarities of dairy management practices in the Alps based on Alpine transhumance and cooperative use of mountain pastures. As the source of the outbreak strain remains unknown, the findings of this study underscore the importance of continuous surveillance by monitoring further spread and persistence of M

  7. Detection and identification of mycoplasmas by amplification of rDNA.

    PubMed

    Blanchard, A; Gautier, M; Mayau, V

    1991-06-01

    Alignment of published 16S rRNA sequences allowed the definition of a pair of oligonucleotides suitable for polymerase chain reaction (PCR). Using this pair of PCR primers, several mycoplasmas including the four human parasites Mycoplasma genitalium, M. hominis, M. salivarium and M. orale were detected. This DNA amplification was restricted to species of the genus Mycoplasma while no cross-reaction was observed with DNA from other bacteria and eukaryotic cells. Subsequent analysis of amplified products by either specific oligonucleotide hybridization or dideoxy sequencing specified the identity of the detected mycoplasmas. This method offers a highly discriminating and sensitive assay for the direct detection and identification of these microorganisms without the need for prior cultivation.

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

    PubMed

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

    2016-10-20

    Fluorescence expression tools for stable and innocuous whole mycoplasma cell labelling have been developed. A Tn4001-derivative mini-transposon affording unmarked, stable mutagenesis in mycoplasmas was modified to allow the constitutive, high-level expression of mCherry, mKO2 and mNeonGreen. These tools were used to introduce the respective fluorescent proteins as chromosomal tags in the phylogenetically distant species Mycoplasma mycoides subsp. mycoides and Mycoplasma bovis. The production, selection and characterisation of fluorescent clones were straightforward and resulted in the unprecedented observation of red and green fluorescent mycoplasma colonies in the two species, with no apparent cytotoxicity. Equivalent fluorescence expression levels were quantified by flow cytometry in both species, suggesting that these tools can be broadly applied in mycoplasmas. A macrophage infection assay was performed to assess the usefulness of mNeonGreen-expressing strains for monitoring mycoplasma infections, and notably cell invasion. The presence of fluorescent mycoplasmas inside live phagocytic cells was detected and quantified by flow cytometry and corroborated by confocal microscopy, which allowed the identification of individual mycoplasmas in the cytoplasm of infected cells. The fluorescence expression tools developed in this study are suitable for host-pathogen interaction studies and offer innumerable perspectives for the functional analysis of mycoplasmas both in vitro and in vivo. PMID:27497759

  9. In vitro susceptibilities of Mycoplasma genitalium to antibiotics.

    PubMed Central

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

    1992-01-01

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

  10. Microbiological and Molecular Characterization of Staphylococcus hominis Isolates from Blood

    PubMed Central

    Mendoza-Olazarán, Soraya; Morfin-Otero, Rayo; Rodríguez-Noriega, Eduardo; Llaca-Díaz, Jorge; Flores-Treviño, Samantha; González-González, Gloria Ma; Villarreal-Treviño, Licet; Garza-González, Elvira

    2013-01-01

    Background Among Coagulase-Negative Staphylococci (CoNS), Staphylococcus hominis represents the third most common organism recoverable from the blood of immunocompromised patients. The aim of this study was to characterize biofilm formation, antibiotic resistance, define the SCCmec (Staphylococcal Chromosomal Cassette mec) type, and genetic relatedness of clinical S. hominis isolates. Methodology S. hominis blood isolates (n = 21) were screened for biofilm formation using crystal violet staining. Methicillin resistance was evaluated using the cefoxitin disk test and the mecA gene was detected by PCR. Antibiotic resistance was determined by the broth microdilution method. Genetic relatedness was determined by pulsed-field gel electrophoresis (PFGE) and SCCmec typed by multiplex PCR using two different methodologies described for Staphylococcus aureus. Results Of the S. hominis isolates screened, 47.6% (10/21) were categorized as strong biofilm producers and 23.8% (5/21) as weak producers. Furthermore, 81% (17/21) of the isolates were methicillin resistant and mecA gene carriers. Resistance to ampicillin, erythromycin, and trimethoprim was observed in >70% of isolates screened. Each isolate showed a different PFGE macrorestriction pattern with similarity ranging between 0–95%. Among mecA-positive isolates, 14 (82%) harbored a non-typeable SCCmec type: eight isolates were not positive for any ccr complex; four contained the mec complex A ccrAB1 and ccrC, one isolate contained mec complex A, ccrAB4 and ccrC, and one isolate contained the mec complex A, ccrAB1, ccrAB4, and ccrC. Two isolates harbored the association: mec complex A and ccrAB1. Only one strain was typeable as SCCmec III. Conclusions The S. hominis isolates analyzed were variable biofilm producers had a high prevalence of methicillin resistance and resistance to other antibiotics, and high genetic diversity. The results of this study strongly suggested that S. hominis isolates harbor new SCCmec

  11. Blastocystis hominis in animals: incidence of four serogroups.

    PubMed

    König, G; Müller, H E

    1997-10-01

    In toto, 520 faecal samples from mammals, birds, reptiles, amphibians, fish, and snails were investigated (see Table 1). 91 strains of Blastocystis hominis could be isolated by culture. However, only 48 of them were suitable for axenisation. 96 percent of samples belonged to four serogroups detected in humans but two strains, one from a pig and another from a duck, could not be classified, suggesting the existence of one or two further serogroups. While humans showed mainly serogroups I and II, pigs harboured serogroups III and IV. Four serogroups were isolated from monkeys. The question whether the genus Blastocystis consists of one or more species is discussed. PMID:9361389

  12. Adherence of Mycoplasma hyopneumoniae to cell monolayers.

    PubMed

    Zielinski, G C; Young, T; Ross, R F; Rosenbusch, R F

    1990-03-01

    This work was an attempt to develop an in vitro adherence model for Mycoplasma hyopneumoniae, using monolayers of human and porcine lung fibroblasts and porcine kidney cells. Mycoplasma hyopneumoniae grown in Friis mycoplasma broth was radiolabeled with 35[S]-methionine, washed, concentrated, and inoculated on the monolayers. After 15 minutes of centrifugation to facilitate adherence, monolayers were washed 3 times, dissolved with 0.1N NaOH, and suspended in scintillation liquid, and the radioactivity was determined in a liquid scintillation counter. Adherence, measured as a percentage of counts added, varied according to the mycoplasma strain and the cell line used. Comparison of strains J, 144L, and 232 of M hyopneumoniae revealed 7.5 +/- 5.9, 31.9 +/- 13, and 9.6 +/- 5% adherence to porcine kidney cells, respectively. Slightly different, but proportionally the same relationships were obtained with swine or human fibroblasts. Adherence was decreased slightly by repeated washings of the mycoplasma-treated cell monolayers; however, a plateau was reached, indicating irreversibility of the adherence process. Pretreatment of cell monolayers with nonlabeled organisms substantially blocked adherence by labeled organisms. Dilution of labeled organisms resulted in an increased proportion adhering. Therefore, it appears that the adherence was a receptor-dependent event. Treatment of the mycoplasmas with trypsin prior to the inoculation of monolayers resulted in a marked reduction in adherence. Treatment of the mycoplasmas with hyperimmune swine serum against M hyopneumoniae or normal swine serum resulted in 80 to 90% reduction of adherence; however, no inhibition occurred when mycoplasmas were treated with purified IgG from the hyperimmune serum.

  13. Mycoplasmas and ovine keratoconjunctivitis.

    PubMed

    Jones, G E; Foggie, A; Sutherland, A; Harker, D B

    1976-08-21

    The clinical course of an outbreak of keratoconjunctivitis in housed lambs and their dams was followed. Signs were transient generally and became severe in only a small proportion of lambs. The outbreak became most obvious when the lambs were 46 to 55 days old, when 46.9 per cent were affected. Mycoplasma conjunctivae isolations, confirmed by comparison with the type strain by biochemical and serological reactions, increased to 62.1 per cent of all eyes swabbed, but no correlation could be demonstrated between presence of the organism and clinical status. The reasons for this are discussed. Mycoplasma ovipneumoniae was also recovered from the eyes of a small number of lambs. Instillation of a broth culture of M conjunctivae into the conjunctival sacs of four hoggs produced a transient keratoconjunctivitis similar to that observed in the field, but no effect was observed in animals inoculated intravenously. M conjunctivae may therefore be the aetiological agent of non-follicular infectious ovine keratoconjunctivitis, although further work in gnotobiotic or specific pathogen free lambs is required to establish the fact beyond doubt.

  14. Atypical mycoplasmas from sheep in Great Britain and Australia identified as Mycoplasma ovipneumoniae.

    PubMed

    Leach, R H; Cottew, G S; Andrews, B E; Powell, D G

    1976-05-01

    Representative strains of "lacy-colony" mycoplasmas isolated from sheep in Great Britain and Victoria (Australia) were classified as M ovipneumoniae following comparison with strain Y98 of this species. The taxonomic description of M ovipneumoniae is extended and Y98 is proposed as the type strain. A brief description is given of the isolation of M ovipneumoniae from sheep in East Anglia.

  15. [The significance of genital mycoplasmas in the etiology of puerperal endometritis].

    PubMed

    Nikonov, A P; Ankirskaia, A S; Nisilevich, V F

    1993-01-01

    The rate of genital Mycoplasma isolation from the uterine cavity was studied in 147 puerperae (80 ones with a normal course of the puerperium and 67 with acute postpartum endometritis). Mycoplasma were isolated from the metroaspirate in 11.3% of puerperae in whom the postpartum period ran a normal course; M. urealyticum were found in 8.8% and M. hominis in 2.5% of cases. In endometritis Mycoplasma were isolated from the infection focus 2.5 times more often, i.e. from 28.4% of patients with postpartum endometritis (M. urealyticum were detected in 9.0% and M. hominis in 19.4% of cases). Mycoplasma were the sole agents of endometritis in 9.0% of patients. Hysteroscopic and morphologic studies helped verify the contribution of genital Mycoplasma to the development of acute postpartum endometritis in 19.4% of the patients. Therefore, virtually every fifth patient with postpartum endometritis was in need of purposeful antimycoplasma therapy with tetracycline. PMID:8048680

  16. ULTRASTRUCTURE OF MYCOPLASMA SPECIES

    PubMed Central

    Domermuth, C. H.; Nielsen, M. H.; Freundt, E. A.; Birch-Andersen, A.

    1964-01-01

    Domermuth, C. H. (Statens Seruminstitut, Copenhagen, Denmark), M. H. Nielsen, E. A. Freundt, and A. Birch-Andersen. Ultrastructure of Mycoplasma species. J. Bacteriol. 88:727–744. 1964.—The ultrastructure of 19 strains (15 species) of Mycoplasmatales grown on solid medium was studied with the aid of an electron microscope. The cells possessed a triple-layered limiting membrane 75 to 100 A thick. This membrane appeared to be symmetrical in some strains and asymmetrical in others. An electron-dense material found in close contact with the cell surface was tentatively interpreted to be a capsular substance. Ribosomes and strands of nuclear material were observed in the cytoplasm of cells of all strains. Ribosomes observed in the JA strain of M. gallisepticum were frequently arranged in a regular geometric pattern of characteristic appearance. Dense inclusions sometimes limited by triple-layered membranes (possibly developing elementary bodies), as well as membrane-surrounded vesicles, were observed in the cytoplasm of cells of some strains. Images PMID:14208513

  17. Bacterial Decolorization of Textile Azo Dye Acid Orange by Staphylococcus hominis RMLRT03

    PubMed Central

    Singh, Rajat Pratap; Singh, Pradeep Kumar; Singh, Ram Lakhan

    2014-01-01

    A bacterial strain RMLRT03 with ability to decolorize textile dye Acid Orange dye was isolated from textile effluent contaminated soil of Tanda, Ambedkar Nagar, Uttar Pradesh (India). The decolorization studies were performed in Bushnell and Haas medium (BHM) amended with Acid Orange dye. The bacterial strain was identified as Staphylococcus hominis on the basis of 16S rDNA sequence. The bacterial strain exhibited good decolorization ability with glucose and yeast extract supplementation as cosubstrate in static conditions. The optimal condition for the decolorization of Acid Orange dye by Staphylococcus hominis RMLRT03 strain were at pH 7.0 and 35°C in 60 h of incubation. The bacterial strain could tolerate high concentrations of Acid Orange dye up to 600 mg l-1. The high decolorizing activity under natural environmental conditions indicates that the bacterial strain has practical application in the treatment of dye containing wastewaters. PMID:25253925

  18. Bacterial Decolorization of Textile Azo Dye Acid Orange by Staphylococcus hominis RMLRT03.

    PubMed

    Singh, Rajat Pratap; Singh, Pradeep Kumar; Singh, Ram Lakhan

    2014-05-01

    A bacterial strain RMLRT03 with ability to decolorize textile dye Acid Orange dye was isolated from textile effluent contaminated soil of Tanda, Ambedkar Nagar, Uttar Pradesh (India). The decolorization studies were performed in Bushnell and Haas medium (BHM) amended with Acid Orange dye. The bacterial strain was identified as Staphylococcus hominis on the basis of 16S rDNA sequence. The bacterial strain exhibited good decolorization ability with glucose and yeast extract supplementation as cosubstrate in static conditions. The optimal condition for the decolorization of Acid Orange dye by Staphylococcus hominis RMLRT03 strain were at pH 7.0 and 35°C in 60 h of incubation. The bacterial strain could tolerate high concentrations of Acid Orange dye up to 600 mg l(-1). The high decolorizing activity under natural environmental conditions indicates that the bacterial strain has practical application in the treatment of dye containing wastewaters.

  19. Actinomyces hominis sp. nov., isolated from a wound swab.

    PubMed

    Funke, Guido; Englert, Ralf; Frodl, Reinhard; Bernard, Kathryn A; Stenger, Steffen

    2010-07-01

    A coryneform bacterium (strain 1094(T)) was isolated from a wound swab taken from an 89-year-old female patient. Chemotaxonomic investigations suggested that this bacterium was related to the genera Actinomyces, Arcanobacterium and Actinobaculum. Phylogenetic analysis of 16S rRNA gene sequences showed that strain 1094(T) was most closely related to Actinomyces europaeus CCUG 32789 A(T) (94.3 % similarity). Phenotypically, the isolate could be separated from its closest phylogenetic neighbours on the basis of being positive for catalase, CAMP reaction, acid phosphatase, N-acetyl-beta-glucosaminidase and raffinose fermentation. Based on the data presented, it is proposed that strain 1094(T) should be classified in a novel species, Actinomyces hominis sp. nov. The type strain is 1094(T) (=CCUG 57540(T) =DSM 22168(T)).

  20. Ultrastructural and cytochemical studies on anionic surface sites of Mycoplasma membranes.

    PubMed

    Schiefer, H G; Krauss, H; Brunner, H; Gerhardt, U

    1977-02-01

    Negatively charged, chemical groups on the surfaces of mycoplasma membranes were studied in the electron microscope by staining with positively charged ferric oxide hydrosols in the acetic acid (AI-reagent) or propanoic acid (PI-reagent), respectively. The technique of thin sectioning was used. With the AI-reagent, no significant binding of the ferric oxide particles was observed on the mycoplasma membranes. With the lipophilic PI-reagent, the membrane surfaces of M. mycoides subsp. capri, M. mycoides subsp. mycoides, M. gallisepticum, and Acholeplasma laidlawii were heavily stained with focal aggregates of iron granules. M. hominis did not bind the label. Prior chemical and enzymatic treatments indicate that the cationic ferric oxide particles in propanoic acid interact with negatively charged, presumably lipid phosphate groups exposed on the surfaces of mycoplasma membranes.

  1. Detection of new mutations conferring resistance to linezolid in glycopeptide-intermediate susceptibility Staphylococcus hominis subspecies hominis circulating in an intensive care unit.

    PubMed

    Sorlozano, A; Gutierrez, J; Martinez, T; Yuste, M E; Perez-Lopez, J A; Vindel, A; Guillen, J; Boquete, T

    2010-01-01

    Glycopeptides and linezolid are the most widely used antibiotics to treat infections by methicillin-resistant Staphylococcus spp. We report the presence of various isolates of methicillin-resistant S. hominis subsp. hominis with resistance to linezolid and reduced susceptibility to glycopeptides. We studied ten blood culture isolates of S. hominis subsp. hominis from nine patients admitted to our hospital. Etest was used to study susceptibility to antibiotics commonly prescribed against staphylococci. Domain V region of the 23S rRNA gene was amplified and sequenced to detect possible mutations that confer resistance to linezolid. Pulsed-field gel electrophoresis (PFGE) was used for the clonality study of isolates. All isolates were resistant to oxacillin, gentamicin, levofloxacin, cotrimoxazole, and linezolid, and susceptible to tigecycline and daptomycin. Nine of the isolates were resistant to erythromycin and clindamycin, and showed heterogeneous resistance to glycopeptides. C2190T, G2603T, and G2474T mutations were detected in domain V of the 23S rRNA gene. PFGE showed the presence of two different clones. This report alerts to the possible appearance of clinical strains of methicillin-resistant staphylococci with intermediate resistance to glycopeptides, resistance to linezolid, and multiple resistance to other second-line antibiotics.

  2. In Vitro Cell Invasion of Mycoplasma gallisepticum

    PubMed Central

    Winner, Florian; Rosengarten, Renate; Citti, Christine

    2000-01-01

    The ability of the widespread avian pathogen Mycoplasma gallisepticum to invade cultured human epithelial cells (HeLa-229) and chicken embryo fibroblasts (CEF) was investigated by using the gentamicin invasion assay and a double immunofluorescence microscopic technique for accurate localization of cell-associated mycoplasmas. The presence of intracellular mycoplasmas in both cell lines was clearly demonstrated, with organisms entering the eukaryotic cells within 20 min. Internalized mycoplasmas have the ability to leave the cell, but also to survive within the intracellular space over a 48-h period. Frequencies of invasion were shown to differ between the two cell lines, but were also considerably dependent on the mycoplasma input population. Of the prototype strain R, a low-passage population in artificial medium, Rlow, was capable of active cell invasion, while a high-passage population, Rhigh, showed adherence to but nearly no uptake into HeLa-229 and CEF. By passaging Rlow and Rhigh multiple times through HeLa-229 cells, the invasion frequency was significantly increased. Taken together, these findings demonstrate that M. gallisepticum has the capability of entering nonphagocytic host cells that may provide this pathogen with the opportunity for resisting host defenses and selective antibiotic therapy, establishing chronic infections, and passing through the respiratory mucosal barrier to cause systemic infections. PMID:10858241

  3. Sequence analysis of 16S rRNA from mycoplasmas by direct solid-phase DNA sequencing.

    PubMed Central

    Pettersson, B; Johansson, K E; Uhlén, M

    1994-01-01

    Automated solid-phase DNA sequencing was used for determination of partial 16S ribosomal DNA sequences of mycoplasmas. The sequence information was used to establish phylogenetic relationships of 11 different mycoplasmas whose 16S rRNA sequences had not been determined earlier. A biotinylated fragment corresponding to positions 344 to 939 in the Escherichia coli sequence was generated by PCR. The PCR product was immobilized onto streptavidin-coated paramagnetic beads, and direct sequencing was performed in both directions. One previously unclassified avian mycoplasma was found to belong to the Mycoplasma lipophilum cluster of the hominis group. Microheterogeneities were discovered in the rRNA operons of Mycoplasma mycoides subsp. mycoides (SC type), confirming the existence of two different rRNA operons. The 16S rRNA sequence of M. mycoides subsp. capri was identical to that of M. mycoides subsp. mycoides (type SC), except that no microheterogeneities were revealed. Furthermore, automated solid-phase DNA sequencing was used to identify a mycoplasmal contamination of a cell culture as Mycoplasma hyorhinis, which proved to be very difficult by conventional methods. The results suggest that the direct solid-phase DNA sequencing procedure is a powerful tool for identification of mycoplasmas and is also useful in taxonomic studies. Images PMID:7521158

  4. Genetic and serological analysis of the immunogenic 67-kDa lipoprotein of Mycoplasma sp. bovine group 7.

    PubMed

    Frey, J; Cheng, X; Monnerat, M P; Abdo, E M; Krawinkler, M; Bölske, G; Nicolet, J

    1998-01-01

    The gene encoding a lipoprotein of 67 kDa, named P67, was cloned from Mycoplasma sp. bovine group 7 strain PG50 and expressed in Escherichia coli K12. Analysis of the amino acid sequence derived from the DNA sequence of the P67 gene revealed a typical prokaryotic signal peptidase II membrane lipoprotein lipid attachment site and a transmembrane structure domain in the leader sequence at the amino-terminal end of the protein. Protein P67 showed 91% identical amino acid residues to the lipoprotein P72 of Mycoplasma mycoides subsp. mycoides small colony type (SC) and 53% identical amino acid residues to a peptide of an unassigned gene on the genome of Mycoplasma capricolum subsp. capricolum. Antibodies made against recombinant P67 reacted with a 67-kDa protein in all Mycoplasma sp. bovine group 7 strains tested and also, to some extent, with P72 of Mycoplasma mycoides subsp. mycoides SC. The gene encoding P67 was present in all strains of Mycoplasma sp. bovine group 7 analysed, but not in other Mycoplasma sp. of the "mycoides cluster" and not in the phylogenetically related Mycoplasma putrefaciens. PCR and restriction fragment analysis revealed that the gene of P67 is conserved in all strains of Mycoplasma sp. bovine group 7. A specific PCR reaction based on the P67 gene sequence enabled rapid identification of strains belonging to Mycoplasma sp. bovine group 7.

  5. 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

  6. Isolate resistance of Blastocystis hominis to metronidazole.

    PubMed

    Haresh, K; Suresh, K; Khairul Anus, A; Saminathan, S

    1999-04-01

    Isolates of Blastocystis hominis from infected immigrant workers from Indonesia, Bangladesh and infected individuals from Singapore and Malaysia were assessed for growth pattern and degree of resistance to different concentrations of metronidazole. Viability of the cells was assessed using eosin-brillian cresyl blue which stained viable cells green and nonviable cells red. The Bangladeshi and Singaporean isolates were nonviable even at the lowest concentration of 0.01 mg/ml, whereas 40% of the initial inoculum of parasites from the Indonesian isolate at day one were still viable in cultures with 1.0 mg/ml metronidazole. The study shows that isolates of B. hominis of different geographical origin have different levels of resistance to metronidazole. The search for more effective drugs to eliminate th parasite appears inevitable, especially since surviving parasites from metronidazole cultures show greater ability to multiply in subcultures than controls. PMID:10357863

  7. Pacemaker lead endocarditis caused by Staphylococcus hominis.

    PubMed

    Sunbul, Mustafa; Demirag, Mustafa Kemal; Yilmaz, Ozcan; Yilmaz, Hava; Ozturk, Recep; Leblebicioglu, Hakan

    2006-05-01

    Infective endocarditis related to pacemaker is a rare but serious condition in permanent venous tracing. A 65-year-old man was admitted to the hospital with high fever and chills. A DDD pacemaker had been implanted via the right subclavian vein because of sick sinus syndrome 6 years earlier. Transesophageal echocardiogram identified an oscillating round hyperechoic mass with a stalk near the tricuspid valve. Blood cultures grew Staphylococcus hominis. The patient was treated with antibiotics and operated on after the acute phase of the illness had subsided. We hereby report a case of lead endocarditis caused by S. hominis in a patient with pacemaker, which has been rarely reported in the English literature.

  8. Revisiting the reference genomes of human pathogenic Cryptosporidium species: reannotation of C. parvum Iowa and a new C. hominis reference

    PubMed Central

    Isaza, Juan P.; Galván, Ana Luz; Polanco, Victor; Huang, Bernice; Matveyev, Andrey V.; Serrano, Myrna G.; Manque, Patricio; Buck, Gregory A.; Alzate, Juan F.

    2015-01-01

    Cryptosporidium parvum and C. hominis are the most relevant species of this genus for human health. Both cause a self-limiting diarrhea in immunocompetent individuals, but cause potentially life-threatening disease in the immunocompromised. Despite the importance of these pathogens, only one reference genome of each has been analyzed and published. These two reference genomes were sequenced using automated capillary sequencing; as of yet, no next generation sequencing technology has been applied to improve their assemblies and annotations. For C. hominis, the main challenge that prevents a larger number of genomes to be sequenced is its resistance to axenic culture. In the present study, we employed next generation technology to analyse the genomic DNA and RNA to generate a new reference genome sequence of a C. hominis strain isolated directly from human stool and a new genome annotation of the C. parvum Iowa reference genome. PMID:26549794

  9. Characteristics of Blastocystis hominis infection in a Turkish university hospital.

    PubMed

    Ozçakir, Olcay; Güreser, Semra; Ergüven, Sibel; Yilmaz, Yakut Akyön; Topaloğlu, Rezzan; Hasçelik, Gülşen

    2007-01-01

    In order to determine characteristics of Blastocystis (B.) hominis infection; 770 individuals' stool specimens were examined both by simple and concentration techniques and stained with iodine solution and trichrome in the Parasitology Laboratory of Hacettepe University Faculty of Medicine, Turkey. Among the examined 770 specimens, B. hominis was detected in 94 (12.2%). B. hominis was the most common intestinal parasite among the study group. It was mostly detected with Dientamoeba fragilis. Among the groups the incidence of B. hominis in allergic patients was higher than controls. Among the immunosuppressed patients, B. hominis was detected significantly higher in patients who had solid tumours. Of the 48 individuals who had only B. hominis in their stool the most common symptom was abdominal pain. Concentration technique with trichrome stain was more sensitive than simple smear with lugol solution for the detection of B. hominis. Studies with more patients must be planed to understand the B. hominis infection in solid tumour patients and coexistence of B. hominis and D. fragilis. PMID:18224616

  10. Role of Vpma phase variation in Mycoplasma agalactiae pathogenesis

    PubMed Central

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

    2015-01-01

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

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

    PubMed Central

    Rocha, Eduardo P. C.; Blanchard, Alain

    2002-01-01

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

  12. Fecal-oral transmission of the cyst form of Blastocystis hominis in rats.

    PubMed

    Yoshikawa, Hisao; Yoshida, Kumi; Nakajima, Ayumi; Yamanari, Kimie; Iwatani, Satoru; Kimata, Isao

    2004-12-01

    The infectivity of two Blastocystis hominis strains, RN94-9 and NIH:1295:1, was examined in 3-week-old SPF Wistar rats. The NIH:1295:1 strain, originally isolated from a guinea pig, was only able to infect rats via intracecal inoculation of the cultured organisms, while the RN94-9 strain, originally isolated from a laboratory rat, was able to infect rats by oral inoculation of the cultures due to the presence of a cystic form in the in vitro culture. Since many cysts were discharged in the feces of the infected rats, the infectivity of the concentrated cysts was compared between the two strains. Successful oral infection was observed in rats inoculated with 1 x 10(2)-1 x 10(6) cysts of the RN94-9 and NIH:1295:1 strains. The infectivity of the ten cysts varied in the three experiments of ten rats, being 20-100% and 30-100% in the RN94-9 and NIH:1295:1 strains, respectively. When an uninfected normal rat was housed with five experimentally inoculated rats, the normal rat became infected, demonstrating the fecal-oral transmission of the cyst form of this parasite. These results show that the Wistar rat is an ideal host for the propagation of strains RN94-9 and NIH:1295:1 of B. hominis, and demonstrate that the cyst form is the only transmissible form of this parasite. PMID:15480786

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    1986-01-01

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

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

    PubMed Central

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

    1986-01-01

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

  17. 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...

  18. Mycoplasma infections of plants.

    PubMed

    Bove, J M

    1981-07-01

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

  19. [Differentiation of glucidolytic mycoplasmas isolated from goats by the API 50 CH system and electrophoresis].

    PubMed

    Richard, Y; Favier, C; Oudar, J

    1991-01-01

    Carbohydrate metabolism of 62 glucidolytic strains of mycoplasma belonging to 4 species (M ovipneumoniae, M putrefaciens, M mycoides, M capricolum) has been studied using the API 50 Ch system for bacterial identification. This microtechnique and colony aspect were relevant in distinguishing M ovipneumoniae and M putrefaciens from the group M mycoides and M capricolum isolated from goats, but still presented a lack of specificity in distinguishing M mycoides from M capricolum. Similar results were obtained when the mycoplasma strains were tested by electrophoresis.

  20. The comparison and characterisation of glycolytic mycoplasmas isolated from the respiratory tract of sheep.

    PubMed

    Jones, G E; Foggie, A; Mould, D L; Livitt, S

    1976-02-01

    Nine strains of glycolytic mycoplasmas isolated from the respiratory tract of apparently healthy sheep, pneumonic sheep and sheep with pulmonary adenomatosis (SPA) were compared with a Queensland strain (Y98) of Mycoplasma ovipneumoniae. All strains were very similar in their reactions in 14 biochemical tests and in their sensitivities to optochin, digitonin, sodium polyanethol sulphonate, and 11 antibiotics. Polyacrylamide-gel electrophoresis and serological cross-reactions by the agar-gel double diffusion, metabolic inhibition (MI) and growht-inhibition (GI) tests also showed that all strains could be classified as M. ovipneumoniae. The MI and GI tests, however, showed considerable intraspecific differences among strains, with apparent polarisation of SPA strains and non-SPA strains at opposite ends of the antigenic spectrum. Two representative strains were tested by the MI test against antisera to 39 mycoplasma species or serogroups, with negative results.

  1. Unusually low prevalence of Mycoplasma genitalium in urine samples from infertile men and healthy controls: a prevalence study

    PubMed Central

    Plecko, Vanda; Zele-Starcevic, Lidija; Tripkovic, Vesna; Skerlev, Mihael; Ljubojevic, Suzana; Plesko, Sanja; Marekovic, Ivana; Jensen, Jorgen Skov

    2014-01-01

    Objective To detect Mycoplasma genitalium in urine samples of infertile men and men without any signs of infection in order to investigate whether M. genitalium and other genital mycoplasmas (Mycoplasma hominis and Ureaplasma spp) are found more often in urine samples of infertile men than in asymptomatic controls and to determine resistance to macrolides. Methods The study included first void urine samples taken from 145 infertile men and 49 men with no symptoms of urethritis. M. genitalium, Chlamydia trachomatis and Neisseria gonorrhoeae were detected by commercial PCR. Trichomonas vaginalis was detected by microscopy and culture. M. hominis and Ureaplasma spp were detected by culture. M. genitalium was detected by in-house conventional and real-time PCR. Results Two M. genitalium positive samples were found among samples obtained from infertile men. All asymptomatic men were M. genitalium negative. Macrolide resistance was not found in either of the two positive samples. Conclusions In comparison with reported data, an unusually low prevalence of M. genitalium was found in infertile men. The reasons for this unexpected result are not known; possibly, local demographic and social characteristics of the population influenced the result. Further studies to investigate M. genitalium in infertile and other groups of patients are needed. PMID:25157184

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

    PubMed

    Purswell, J L; Evans, J D; Branton, S L

    2011-09-01

    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 the vaccine suspension droplet trajectory. In this study, a total of 48 Hy-Line W-36 males were placed individually in isolation units following eye-drop application of gradient levels (1 x, 10(-1) x, 10(-2) x, 10(-3) x, 10(-4) x, 10(-5) x, 10(-6) x, and unvaccinated control) of the MG vaccine. The determined titer associated with a 1 x dose was 2 x 10(6) colony-forming units/dose. Serologic response was assessed weekly following vaccination via serum plate agglutination (SPA) for weeks one through seven postvaccination (p.v.). In addition, immunologic response was assessed at 5, 6, and 7 wk p.v. via MG enzyme-linked immunosorbent assay (ELISA). As indicated by SPA analyses, a 1 x dose of vaccine resulted in 100% seroconversion, and dose levels of 10(-1) x and 10(-2) x resulted in 75% and 37.5% seroconversion, respectively, at 6 wk p.v. The MG ELISA results at 6 wk p.v. demonstrated immunologic responses in 100%, 57.1%, and 28.6% of the 1 x, 10(-1) x, and 10(-2) x dosed birds, respectively. The lower dosage levels of 10(-3) x, 10(-4) x, 10(-5) x, and 10(-6) x did not elicit a response from any bird at 6 wk p.v. Utilizing the SPA data, a logistic regression model was used to determine the relationship between dosage level and seroconversion rate (R2 = 0.999 with a standard error of prediction of 1.6%). The model predicted a required effective dosage of 0.26 x for 90% seroconversion at 6 wk p.v. under test conditions.

  3. Membrane proteins of Mycoplasma bovis and their role in pathogenesis.

    PubMed

    Adamu, James Y; Wawegama, Nadeeka K; Browning, Glenn F; Markham, Philip F

    2013-10-01

    Mycoplasma membrane proteins influence cell shape, cell division, motility and adhesion to host cells, and are thought to be integrally involved in the pathogenesis of mycoplasmoses. Many of the membrane proteins predicted from mycoplasma genome sequences remain hypothetical, as their presence in cellular protein preparations is yet to be established experimentally. Recent genome sequences of several strains of Mycoplasma bovis have provided further insight into the potential role of the membrane proteins of this pathogen in colonisation and infection. This review highlights recent advances in knowledge about the influence of M. bovis membrane proteins on the pathogenesis of infection with this species and identifies future research directions for enhancing our understanding of the role of these proteins. PMID:23810376

  4. Experimental Sarcocystis hominis infection in a water buffalo (Bubalus bubalis).

    PubMed

    Chen, X W; Zuo, Y X; Hu, J J

    2003-04-01

    A water buffalo (Bubalus bubalis) was fed 5.0 x 10(5) Sarcocystis hominis sporocysts from a human volunteer who had ingested S. hominis cysts from naturally infected cattle. A necropsy was performed on the buffalo 119 days after inoculation, and a large number of microscopic sarcocysts (approximately 5,000/g) were found in skeletal muscles. Ultrastructurally, the sarcocyst wall from buffalo muscles has upright villar protrusions measuring about 5.6 x 0.8 microm with numerous microtubules that run from the base to the apex. Sarcocysts from this buffalo were infective to 2 human volunteers, confirming their identity as S. hominis. Therefore, we believe that buffaloes can act experimentally as the intermediate host for S. hominis.

  5. Isolation and immunological detection of Mycoplasma ovipneumoniae in sheep with atypical pneumonia, and lack of a role for Mycoplasma arginini.

    PubMed

    Lin, Y-C; Miles, R J; Nicholas, R A J; Kelly, D P; Wood, A P

    2008-06-01

    Mycoplasma ovipneumoniae NCTC 10151(T) and four new isolates from UK sheep flocks were compared. Only glucose and pyruvate were used as energy sources by the five strains: glucose was the best energy source for the type strain, pyruvate supported better growth of the new strains. Whole cell protein patterns and antigenic profiles showed high similarity between all five strains. The new isolates fell into two groups in ELISA tests. Serum samples from 30 pneumonic sheep were assessed for M. ovipneumoniae infection and Mycoplasma arginini co-infection. Fourteen (out of 30) serum samples were positive for M. ovipneumoniae both by ELISA and immunoblotting. Twelve antigenic proteins of M. ovipneumoniae were detected in infected serum samples: the antigen patterns were unique, with between one and at least seven occurring in any one sample. All serum samples were designated as negative for M. arginini antibodies by both ELISA and immunoblotting.

  6. The proline-rich P65 protein of Mycoplasma pneumoniae is a component of the Triton X-100-insoluble fraction and exhibits size polymorphism in the strains M129 and FH.

    PubMed Central

    Proft, T; Hilbert, H; Layh-Schmitt, G; Herrmann, R

    1995-01-01

    Previously, we described the identification of a novel Mycoplasma pneumoniae M129 protein, named P65 because of its apparent molecular mass of 65 kDa estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (T. Proft and R. Herrmann, Mol. Microbiol. 13:337-348, 1994). DNA sequence analysis of the P65 open reading frame (orfp65), however, revealed an ORF encoding a protein with a molecular weight of 47,034. This discrepancy can be explained by the unusual amino acid composition of this protein. According to the deduced amino acid sequence, the N-terminal half of P65 contains several penta- and hexapeptides (DPNAY and DPNQAY) forming a proline-rich acidic domain. Secondary-structure predictions indicated beta-sheets and turns within that region, suggesting an extended and rigid conformation. Near the C terminus of P65 the tripeptide Arg-Gly-Asp (RGD) was found. This motif is known to play an important role in binding of extracellular matrix proteins to integrins. P65 could be located exclusively to the Triton X-100-insoluble cell fraction. The results of immunofluorescence microscopy and of immunoadsorption experiments indicated that P65 carries surface-exposed regions. Mild treatment of whole cells with proteases resulted in cleavage of a limited amount of P65 molecules, suggesting either that only a small percentage of P65 molecules are exposed on the surface or that protease cleavage is hampered by a compact protein conformation or by binding of an unknown component to P65. P65 exhibits size polymorphism in M. pneumoniae M129 and FH. This is caused by an intragenetic duplication of a 54-bp sequence within the FH orfp65. As a consequence, the number of DPNAY pentapeptides increased from 9 to 12 repeats in the FH strain. PMID:7768845

  7. Identification of lipoprotein MslA as a neoteric virulence factor of Mycoplasma gallisepticum.

    PubMed

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

    2010-08-01

    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 R(low), 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 R(low) and S6. We examined the virulence of an R(low) Delta MGA0674 mutant (P1H9) in vivo and observed reduced recovery and attenuated virulence in the tracheas of experimentally infected chickens. The virulence of two additional R(low) Delta 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

  8. Mycoplasmas isolated from the respiratory tract of horses.

    PubMed Central

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

    1975-01-01

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

  9. Alterations in the metabolism of hamster tracheas in organ culture after infection by virulent Mycoplasma pneumoniae.

    PubMed

    Hu, P C; Collier, A M; Baseman, J B

    1975-04-01

    Exposure of hamster tracheal rings in organ culture to virulent Mycoplasma pneumoniae organisms leads to alterations in macromolecular biosynthesis and metabolic activity of the respiratory epithelial cells. Avirulent organisms derived from the same parent strain do not produce these effects. During the course of infection by virulent mycoplasmas, tracheal rings show an initial increase in [14C]galactose uptake followed by a significant decline as infection progresses which is also accompanied by abnormal processing of galactose as evidenced by amounts of 14CO2 released. Parallel decreases in the rate of [3H]orotic acid and [3H]amino acid uptake are observed. Within 24 h after infection of tracheal rings by virulent mycoplasmas, inhibition of host cell ribonucleic acid and protien synthesis is evident. Ribonucleic acid synthesis in infected cells, analyzed by gel electrophoresis, is reduced by 80% at 48 h and is negligible by 96 h. The course of mycoplasma infection can be interrupted or reversed by erythromycin after the initial mycoplasma-host cell interaction since addition of erythromycin 24 h or earlier after infection prevents the onset of abnormal orotic acid uptake. However, 48 h after infection, rescue of host cells by erythromycin cannot occur and cytopathology becomes evident. These data suggest that mediation of host cell injury requires continued protein synthesis by attached mycoplasmas, and the primary effect of mycoplasma infection on tracheal organ culture may be at a transcriptional or translational level.

  10. Pulse-field electrophoresis indicates full-length Mycoplasma chromosomes range widely in size.

    PubMed Central

    Neimark, H C; Lange, C S

    1990-01-01

    Full-size linear chromosomes were prepared from mycoplasmas by using gamma-irradiation to introduce one (on average) double-strand break in their circular chromosomes. Chromosome sizes were estimated by pulsed-field gel electrophoresis (PFGE) from the mobilities of these full-length molecules relative to DNA size references. Sizes estimated for Ureaplasma urealyticum T960 and 16 Mycoplasma species ranged from 684 kbp (M. hominis) to 1315 kbp (M. iowae). Using this sample, we found no correlation between the mobility of the full-size linear chromosomes and their G + C content. Sizes for A. laidlawii and A. hippikon were within the range expected from renaturation kinetics. PFGE size estimates are in good agreement with sizes determined by other methods, including electron microscopy, an ordered clone library, and summation of restriction fragments. Our estimates also agree with those from renaturation kinetics for both the largest and some of the smallest chromosomes, but in the intermediate size range, renaturation kinetics consistently provides lower values than PFGE or electron microscopy. Our PFGE estimates show that mycoplasma chromosomes span a continual range of sizes, with several intermediate values falling between the previously recognized large and small chromosome size clusters. Images PMID:2216718

  11. 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.

  12. A foodborne outbreak of Cryptosporidium hominis infection.

    PubMed

    Ethelberg, S; Lisby, M; Vestergaard, L S; Enemark, H L; Olsen, K E P; Stensvold, C R; Nielsen, H V; Porsbo, L J; Plesner, A-M; Mølbak, K

    2009-03-01

    Foodborne outbreaks of cryptosporidiosis are uncommon. In Denmark human cases are generally infrequently diagnosed. In 2005 an outbreak of diarrhoea affected company employees near Copenhagen. In all 99 employees were reported ill; 13 were positive for Cryptosporidium hominis infection. Two analytical epidemiological studies were performed; an initial case-control study followed by a cohort study using an electronic questionnaire. Disease was associated with eating from the canteen salad bar on one, possibly two, specific weekdays [relative risk 4.1, 95% confidence interval (CI) 2.1-8.3]. Three separate salad bar ingredients were found to be likely sources: peeled whole carrots served in a bowl of water, grated carrots, and red peppers (in multivariate analysis, whole carrots: OR 2.1, 95% CI 1.1-4.0; grated carrots: OR 2.1, 95% CI 1.2-3.9; peppers: OR 3.3, 95% CI 1.7-6.6). We speculate that a person excreting the parasite may have contaminated the salad buffet.

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

    PubMed

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

    2008-10-15

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

  14. Clinical significance and prevalence of Blastocystis hominis in Van, Turkey

    PubMed Central

    Beyhan, Yunus E.; Yilmaz, Hasan; Cengiz, Zeynep T.; Ekici, Abdurrahman

    2015-01-01

    Objectives: To determine the associated clinical symptoms and prevalence of Blastocystis hominis (B. hominis). Methods: Stool samples of 50,185 patients (26,784 males and 23,401 females) who were received at the Parasitology Laboratory of Yuzuncu Yil University Faculty of Medicine, Van, Turkey in the last 5 years were inspected microscopically using saline and iodine-stained wet-mount preparations. Age, gender, and symptoms of patients were recorded and their significance was evaluated. Results: The prevalence of B. hominis in the total sample was 0.54% (275/50185). Out of 275 infected patients, 143 (52%) were males, and 132 (48%) were female (χ2=0.884; p=0.348). The distribution of B. hominis infection was high in 7-13 aged children (34.9%) (χ2=306.8; p=0.001). Blastocystis was higher among symptomatic patients (70.2%) compared with asymptomatic patients (29.8%) (χ2=107.13; p=0.001). The most frequent clinical symptoms associated with the disease were abdominal pain (27.3%) and diarrhea (19.6%) followed by anorexia, fever, saliva, anal itching, and nausea. Conclusion: Blastocystis hominis is considered a causative agent of human disease in patients with recurrent symptoms. Due to the significant risk for zoonotic transmission, molecular techniques must be used to determine the route and source of infection. PMID:26318472

  15. Pentatrichomonas hominis in laboratory-bred common marmosets

    PubMed Central

    Inoue, Takashi; Hayashimoto, Nobuhito; Yasuda, Masahiko; Sasaki, Erika; Itoh, Toshio

    2015-01-01

    Trichomonadid protozoa have been found in the intestinal tracts of common marmosets (Callithrix jacchus). However, there is little information available on species identification and the pathogenicity of these trichomonads. In this study, we conducted a fecal survey of a common marmoset colony maintained as laboratory animals in Japan and identified the trichomonad species. Screening using a fecal smear examination revealed that 66% (58/88) of the marmosets had trichomonadid trophozoites in their feces. The trichomonads were found in both normal feces (31/49, 63%) and diarrhea (27/39, 69%), with no significant difference in frequency. The protozoa were identified as Pentatrichomonas hominis using morphological characters and the 100% identity of the nucleotide sequence of the partial 18S rRNA gene (297 bp). The intraspecific genetic variability between P. hominis from the marmosets in this study and P. hominis from other reported mammal hosts was ≤1% in the nucleotide sequence, including the internal transcribed spacer (ITS)-1, 5.8S rRNA gene, and ITS-2 (293 bp). P. hominis inhabits the large intestine of various mammalian hosts, including primates, and is considered nonpathogenic. These results suggest that P. hominis is transmitted among marmosets and other mammals but is not a primary cause of bowel disease in marmosets. PMID:26156572

  16. Common occurrence of Cryptosporidium hominis in horses and donkeys.

    PubMed

    Jian, Fuchun; Liu, Aiqin; Wang, Rongjun; Zhang, Sumei; Qi, Meng; Zhao, Wei; Shi, Yadong; Wang, Jianling; Wei, Jiujian; Zhang, Longxian; Xiao, Lihua

    2016-09-01

    Extensive genetic variation is observed within the genus Cryptosporidium and the distribution of Cryptosporidium species/genotypes in humans and animals appears to vary by geography and host species. To better understand the genetic diversity of Cryptosporidium spp. in horses and donkeys, we characterized five horse-derived and 82 donkey-derived Cryptosporidium isolates from five provinces or autonomous regions (Sichuan, Gansu, Henan, Inner Mongolia and Shandong) in China at the species/genotype and subtype levels. Three Cryptosporidium species/genotypes were identified based on the analysis of the SSU rRNA gene, including Cryptosporidium parvum (n=22), the Cryptosporidium horse genotype (n=4), and Cryptosporidium hominis (n=61). The identification of C. hominis was confirmed by sequence analysis of the HSP70 and actin genes. Subtyping using sequence analysis of the 60kDa glycoprotein gene identified 21 C. parvum isolates as subtype IIdA19G1, the four horse genotype isolates as subtypes VIaA15G4 (n=2) and VIaA11G3 (n=2), and the 61 C. hominis isolates as IkA16G1 (n=59) and IkA16 (n=2). The common finding of C. hominis reaffirms the heterogeneity of Cryptosporidium spp. in horses and donkeys and is possibly a reflection of endemic transmission of C. hominis in these animals. Data of the study suggest that horses and donkeys as companion animals may potentially transmit Cryptosporidium infections to humans.

  17. Common occurrence of Cryptosporidium hominis in horses and donkeys.

    PubMed

    Jian, Fuchun; Liu, Aiqin; Wang, Rongjun; Zhang, Sumei; Qi, Meng; Zhao, Wei; Shi, Yadong; Wang, Jianling; Wei, Jiujian; Zhang, Longxian; Xiao, Lihua

    2016-09-01

    Extensive genetic variation is observed within the genus Cryptosporidium and the distribution of Cryptosporidium species/genotypes in humans and animals appears to vary by geography and host species. To better understand the genetic diversity of Cryptosporidium spp. in horses and donkeys, we characterized five horse-derived and 82 donkey-derived Cryptosporidium isolates from five provinces or autonomous regions (Sichuan, Gansu, Henan, Inner Mongolia and Shandong) in China at the species/genotype and subtype levels. Three Cryptosporidium species/genotypes were identified based on the analysis of the SSU rRNA gene, including Cryptosporidium parvum (n=22), the Cryptosporidium horse genotype (n=4), and Cryptosporidium hominis (n=61). The identification of C. hominis was confirmed by sequence analysis of the HSP70 and actin genes. Subtyping using sequence analysis of the 60kDa glycoprotein gene identified 21 C. parvum isolates as subtype IIdA19G1, the four horse genotype isolates as subtypes VIaA15G4 (n=2) and VIaA11G3 (n=2), and the 61 C. hominis isolates as IkA16G1 (n=59) and IkA16 (n=2). The common finding of C. hominis reaffirms the heterogeneity of Cryptosporidium spp. in horses and donkeys and is possibly a reflection of endemic transmission of C. hominis in these animals. Data of the study suggest that horses and donkeys as companion animals may potentially transmit Cryptosporidium infections to humans. PMID:27264727

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  19. 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...

  20. DNA repair in reduced genome: the Mycoplasma model.

    PubMed

    Carvalho, Fabíola Marques; Fonseca, Marbella Maria; Batistuzzo De Medeiros, Sílvia; Scortecci, Kátia Castanho; Blaha, Carlos Alfredo Galindo; Agnez-Lima, Lucymara Fassarella

    2005-11-01

    The occurrence of bacteria with a reduced genome, such as that found in Mycoplasmas, raises the question as to which genes should be enough to guarantee the genomic stability indispensable for the maintenance of life. The aim of this work was to compare nine Mycoplasma genomes in regard to DNA repair genes. An in silico analysis was done using six Mycoplasma species, whose genomes are accessible at GenBank, and M. synoviae, and two strains of M. hyopneumoniae, whose genomes were recently sequenced by The Brazilian National Genome Project Consortium and Southern Genome Investigation Program (Brazil) respectively. Considering this reduced genome model, our comparative analysis suggests that the DNA integrity necessary for life can be primarily maintained by nucleotide excision repair (NER), which is the only complete repair pathway. Furthermore, some enzymes involved with base excision repair (BER) and recombination are also present and can complement the NER activity. The absence of RecR and RecO-like ORFs was observed only in M. genitalium and M. pneumoniae, which can be involved with the conservation of gene order observed between these two species. We also obtained phylogenetic evidence for the recent acquisition of the ogt gene in M. pulmonis and M. penetrans by a lateral transference event. In general, the presence or nonexistence of repair genes is shared by all species analyzed, suggesting that the loss of the majority of repair genes was an ancestral event, which occurred before the divergence of the Mycoplasma species. PMID:16153783

  1. [Update on Dermatobia hominis: South American furuncular myiasis].

    PubMed

    Clyti, E; Pages, F; Pradinaud, R

    2008-02-01

    Furuncular myiasis is an infestation of the skin caused by Dermatobia hominis larvae known as "ver macaque" in French Guyana, "berne" in Brazil, "torsalo" in Colombia, or "human botfly" in English-language literature. It has identical features in man and domestic mammals. The primary lesion consists of a boil-like inflammatory papule with a central punctum exuding a serosanguinous discharge. The respiratory sinus of the D. hominis larvae may be visible through the punctum. Myiasis secondary to D. hominis accounts for 10% of imported tropical dermatosis observed in Paris. Diagnosis of furuncular myiasis should be considered in any patient with a history of travel or residence in an endemic area. Treatment depends mainly on mechanical removal that may be facilitated by injection of lidocaine into the lesion or prior application of a 1% solution of ivermectin.

  2. 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

  3. Multilocus sequence typing and further genetic characterization of the enigmatic pathogen, Staphylococcus hominis.

    PubMed

    Zhang, Liangfen; Thomas, Jonathan C; Miragaia, Maria; Bouchami, Ons; Chaves, Fernando; d'Azevedo, Pedro A; Aanensen, David M; de Lencastre, Herminia; Gray, Barry M; Robinson, D Ashley

    2013-01-01

    Staphylococcus hominis is a commensal resident of human skin and an opportunistic pathogen. The species is subdivided into two subspecies, S. hominis subsp. hominis and S. hominis subsp. novobiosepticus, which are difficult to distinguish. To investigate the evolution and epidemiology of S. hominis, a total of 108 isolates collected from 10 countries over 40 years were characterized by classical phenotypic methods and genetic methods. One nonsynonymous mutation in gyrB, scored with a novel SNP typing assay, had a perfect association with the novobiocin-resistant phenotype. A multilocus sequence typing (MLST) scheme was developed from six housekeeping gene fragments, and revealed relatively high levels of genetic diversity and a significant impact of recombination on S. hominis population structure. Among the 40 sequence types (STs) identified by MLST, three STs (ST2, ST16 and ST23) were S. hominis subsp. novobiosepticus, and they distinguished between isolates from different outbreaks, whereas 37 other STs were S. hominis subsp. hominis, one of which was widely disseminated (ST1). A modified PCR assay was developed to detect the presence of ccrAB4 from the SCCmec genetic element. S. hominis subsp. novobiosepticus isolates were oxacillin-resistant and carriers of specific components of SCCmec (mecA class A, ccrAB3, ccrAB4, ccrC), whereas S. hominis subsp. hominis included both oxacillin-sensitive and -resistant isolates and a more diverse array of SCCmec components. Surprisingly, phylogenetic analyses indicated that S. hominis subsp. novobiosepticus may be a polyphyletic and, hence, artificial taxon. In summary, these results revealed the genetic diversity of S. hominis, the identities of outbreak-causing clones, and the evolutionary relationships between subspecies and clones. The pathogenic lifestyle attributed to S. hominis subsp. novobiosepticus may have originated on more than one occasion.

  4. An improved loop-mediated isothermal amplification assay for the detection of Mycoplasma bovis

    PubMed Central

    HIGA, Yumiko; UEMURA, Ryoko; YAMAZAKI, Wataru; GOTO, Shinya; GOTO, Yoshitaka; SUEYOSHI, Masuo

    2016-01-01

    We improved a loop-mediated isothermal amplification (LAMP) assay permitting sensitive and rapid Mycoplasma bovis detection. A total of 55 bacterial strains were examined in this study, including 33 M. bovis strains, 14 non-M. bovis mycoplasmas and eight non-mycoplasma bacterial strains. M. bovis was successfully detected by the LAMP assay within 60 min without cross-reaction to any other bacteria. Furthermore, a total of 135 nasal swab samples were tested directly using our LAMP assays, the previously reported LAMP assay, conventional PCR assay without pre-culture and comparing standard culture methods. The improved LAMP assay showed sensitivity and specificity of 97.2% and 90.9%, respectively (with a kappa coefficient of 0.8231), and the sensitivity of our revised LAMP assay was increased compared to existing methods. PMID:27109067

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

    PubMed

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

    2014-01-01

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

  6. A sebaceous cyst with a difference: Dermatobia hominis.

    PubMed

    Harbin, L J; Khan, M; Thompson, E M; Goldin, R D

    2002-10-01

    Dermatobia hominis causes furuncular myiasis and is endemic to South America. This report describes a case in a young woman who had recently visited Belize, highlighting the importance of clinical history (including travel history) and close liaison between pathologist and surgeon. PMID:12354816

  7. 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

  8. Motility of Mycoplasma pneumoniae.

    PubMed Central

    Radestock, U; Bredt, W

    1977-01-01

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

  9. Various blood parameters in commercial hens acutely and chronically infected with Mycoplasma gallisepticum and Mycoplasma synoviae.

    PubMed

    Branton, S L; May, J D; Lott, B D; Maslin, W R

    1997-01-01

    Two trials were conducted to study the effects of acute (Trial 1) and chronic (Trial 2) mycoplasma infections on differential leukocyte counts in chickens. The trials initially included either 20 (Trial 1) or 40 (Trial 2) 6-wk-old commercial leghorn chickens negative for antibodies to Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS). Chickens were inoculated with F strain MG (FMG), MS (WVU 1853), or both. One group of chickens remained uninoculated and served as a negative control for both trials. Chickens were housed in fiberglass isolation units from 6 to 10 wk (Trial 1) or 6 to 70 wk of age (Trial 2). Differential leukocyte counts were examined from 6 to 10 wk (Trial 1) or 66 to 70 wk of age (Trial 2) in all chickens. Also, in Trial 2, packed cell volumes (PCVs) and plasma protein values were examined from 66 to 70 wk of age. In the acute study (Trial 1), differential leukocyte counts revealed statistically significant differences (P < 0.05) in heterophil, lymphocyte, monocyte, eosinophil, and basophil values among treatments. In general, the differential counts of FMG- and MS-infected birds were characterized by heterophilia, lymphopenia, monocytosis, eosinopenia, and basopenia. Histopathologic examination of the spleen, liver, kidney, and bone marrow revealed a high degree of lymphoid foci within the spleen and bone marrow of all infected chickens. In the chronic study (Trial 2), no statistically significant differences (P < 0.05) were observed in differential leukocyte counts, PCV, and plasma protein values among treatments. Histopathologic examination of spleen, liver, kidney, and bone marrow did not reveal any difference among treatments.

  10. The effect of antibiotics against bovine mycoplasmas and ureaplasmas.

    PubMed Central

    Truscott, R B; Ruhnke, H L

    1984-01-01

    A combination of lincomycin-spectinomycin-tylosin was tested against several strains of mycoplasmas and acholeplasmas as might be encountered in bovine semen and shown to be effective against them. This combination as well as minocin , rosaramicin, rosoxacin, tiamulin, gentamicin and declomycin were tested in vitro against 58 isolates of ureaplasma from the bovine urogenital tract. The lincomycin-spectinomycin-tylosin combination, minocin , rosaramicin, tiamulin and declomycin were quite active, while rosoxacin and gentamicin were much less active against the test strains. PMID:6232992

  11. 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, Isabella 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.

  12. The occurrence of Mycoplasma phocicerebrale, Mycoplasma phocidae, and Mycoplasma phocirhinis in grey and common seals (Halichoerus grypus and Phoca vitulina) in the United Kingdom.

    PubMed

    Ayling, Roger D; Bashiruddin, Samantha; Davison, Nicholas J; Foster, Geoffrey; Dagleish, Mark P; Nicholas, Robin A J

    2011-04-01

    Following the isolation of Mycoplasma phocicerebrale from the flipper wound of a grey seal (Halichoerus grypus) in Cornwall, UK, surveillance for Mycoplasma species was extended to include other seals rescued or found dead around the UK. Mycoplasma phocicerebrale was frequently detected from the teeth of seals and from infected wounds and respiratory tracts. Mycoplasma phocirhinis, Mycoplasma phocidae, and some unidentified Mycoplasma species were also detected. Mycoplasma phocicerebrale and M. phocidae were the only bacteria consistently identified from the wound infections, but their role in respiratory and other diseases remains unknown, as other bacteria were also isolated from respiratory sites. PMID:21441202

  13. Variable Surface Protein Vmm of Mycoplasma mycoides subsp. mycoides Small Colony Type

    PubMed Central

    Persson, Anja; Jacobsson, Karin; Frykberg, Lars; Johansson, Karl-Erik; Poumarat, François

    2002-01-01

    A variable surface protein, Vmm, of the bovine pathogen Mycoplasma mycoides subsp. mycoides small colony type (M. mycoides SC) has been identified and characterized. Vmm was specific for the SC biotype and was expressed by 68 of 69 analyzed M. mycoides SC strains. The protein was found to undergo reversible phase variation at a frequency of 9 × 10−4 to 5 × 10−5 per cell per generation. The vmm gene was present in all of the 69 tested M. mycoides SC strains and encodes a lipoprotein precursor of 59 amino acids (aa), where the mature protein was predicted to be 36 aa and was anchored to the membrane by only the lipid moiety, as no transmembrane region could be identified. DNA sequencing of the vmm gene region from ON and OFF clones showed that the expression of Vmm was regulated at the transcriptional level by dinucleotide insertions or deletions in a repetitive region of the promoter spacer. Vmm-like genes were also found in four closely related mycoplasmas, Mycoplasma capricolum subsp. capricolum, M. capricolum subsp . capripneumoniae, Mycoplasma sp. bovine serogroup 7, and Mycoplasma putrefaciens. However, Vmm could not be detected in whole-cell lysates of these species, suggesting that the proteins encoded by the vmm-like genes lack the binding epitope for the monoclonal antibody used in this study or, alternatively, that the Vmm-like proteins were not expressed. PMID:12057968

  14. Genital Mycoplasma and Chlamydia trachomatis infections in patients with genital tract infections attending a tertiary care hospital of North India.

    PubMed

    Saigal, Karnika; Dhawan, Benu; Rawre, Jyoti; Khanna, Neena; Chaudhry, Rama

    2016-01-01

    Limited data are available on the prevalence of genital mycoplasmas and Chlamydia trachomatis (CT) among Indian patients with genital tract infections. The objectives of the study were to determine the prevalence of Ureaplasma urealyticum (UU), Mycoplasma hominis (MH), Mycoplasma genitalium (MG), and CT in patients with genital tract infections. The antimicrobial susceptibilities of UU and MH were also assessed. Endocervical swabs/urethral swabs and first void urine samples of patients (n = 164) were collected. UU and MH were detected by culture and multiplex polymerase chain reaction (PCR). MG and CT were identified by PCR. Ureaplasma isolates were further biotyped and serotyped. Antimicrobial susceptibility was done by microbroth dilution method. UU, MH, MG, and CT were detected in 15.2%, 5.4%, 1.2%, and 6% patients, respectively. Ureaplasma parvum serovar 3/14 was the most prevalent. All isolates of UU and MH were uniformly susceptible to doxycycline and josamycin. Routine screening for these pathogens and antimicrobial susceptibility testing is warranted to prevent sequel of infections and formulate treatment guidelines.

  15. Genital Mycoplasma and Chlamydia trachomatis infections in patients with genital tract infections attending a tertiary care hospital of North India.

    PubMed

    Saigal, Karnika; Dhawan, Benu; Rawre, Jyoti; Khanna, Neena; Chaudhry, Rama

    2016-01-01

    Limited data are available on the prevalence of genital mycoplasmas and Chlamydia trachomatis (CT) among Indian patients with genital tract infections. The objectives of the study were to determine the prevalence of Ureaplasma urealyticum (UU), Mycoplasma hominis (MH), Mycoplasma genitalium (MG), and CT in patients with genital tract infections. The antimicrobial susceptibilities of UU and MH were also assessed. Endocervical swabs/urethral swabs and first void urine samples of patients (n = 164) were collected. UU and MH were detected by culture and multiplex polymerase chain reaction (PCR). MG and CT were identified by PCR. Ureaplasma isolates were further biotyped and serotyped. Antimicrobial susceptibility was done by microbroth dilution method. UU, MH, MG, and CT were detected in 15.2%, 5.4%, 1.2%, and 6% patients, respectively. Ureaplasma parvum serovar 3/14 was the most prevalent. All isolates of UU and MH were uniformly susceptible to doxycycline and josamycin. Routine screening for these pathogens and antimicrobial susceptibility testing is warranted to prevent sequel of infections and formulate treatment guidelines. PMID:27166039

  16. Mycoplasma agalactiae MAG_5040 is a Mg2+-Dependent, Sugar-Nonspecific SNase Recognised by the Host Humoral Response during Natural Infection

    PubMed Central

    Cacciotto, Carla; Addis, Maria Filippa; Coradduzza, Elisabetta; Carcangiu, Laura; Nuvoli, Anna Maria; Tore, Gessica; Dore, Gian Mario; Pagnozzi, Daniela; Uzzau, Sergio; Chessa, Bernardo; Pittau, Marco; Alberti, Alberto

    2013-01-01

    In this study the enzymatic activity of Mycoplasma agalactiae MAG_5040, a magnesium-dependent nuclease homologue to the staphylococcal SNase was characterized and its antigenicity during natural infections was established. A UGA corrected version of MAG_5040, lacking the region encoding the signal peptide, was expressed in Escherichia coli as a GST fusion protein. Recombinant GST-MAG_5040 exhibits nuclease activity similar to typical sugar-nonspecific endo- and exonucleases, with DNA as the preferred substrate and optimal activity in the presence of 20 mM MgCl2 at temperatures ranging from 37 to 45°C. According to in silico analyses, the position of the gene encoding MAG_5040 is consistently located upstream an ABC transporter, in most sequenced mycoplasmas belonging to the Mycoplasma hominis group. In M. agalactiae, MAG_5040 is transcribed in a polycistronic RNA together with the ABC transporter components and with MAG_5030, which is predicted to be a sugar solute binding protein by 3D modeling and homology search. In a natural model of sheep and goats infection, anti-MAG_5040 antibodies were detected up to 9 months post infection. Taking into account its enzymatic activity, MAG_5040 could play a key role in Mycoplasma agalactiae survival into the host, contributing to host pathogenicity. The identification of MAG_5040 opens new perspectives for the development of suitable tools for the control of contagious agalactia in small ruminants. PMID:23469065

  17. Identification in methicillin-susceptible Staphylococcus hominis of an active primordial mobile genetic element for the staphylococcal cassette chromosome mec of methicillin-resistant Staphylococcus aureus.

    PubMed

    Katayama, Yuki; Takeuchi, Fumihiko; Ito, Teruyo; Ma, Xiao Xue; Ui-Mizutani, Yoko; Kobayashi, Ichizo; Hiramatsu, Keiichi

    2003-05-01

    We previously reported that the methicillin resistance gene mecA is carried by a novel type of mobile genetic element, SCCmec (staphylococcal cassette chromosome mec), in the chromosome of methicillin-resistant Staphylococcus aureus (MRSA). These elements are precisely excised from the chromosome and integrated into a specific site on the recipient chromosome by a pair of recombinase proteins encoded by the cassette chromosome recombinase genes ccrA and ccrB. In the present work, we detected homologues of the ccr genes in Staphylococcus hominis type strain GIFU12263 (equivalent to ATCC 27844), which is susceptible to methicillin. Sequence determination revealed that the ccr homologues in S. hominis were type 1 ccr genes (ccrA1 and ccrB1) that were localized on a genetic element structurally very similar to SCCmec except for the absence of the methicillin-resistance gene, mecA. This genetic element had mosaic-like patterns of homology with extant SCCmec elements, and we designated it SCC(12263) and considered it a type I staphylococcal cassette chromosome (SCC). The ccrB1 gene identified in the S. hominis strain is the first type 1 ccrB gene discovered to retain its function through the excision process as judged by two criteria: (i) SCC(12263) was spontaneously excised during cultivation of the strain and (ii) introduction of the S. hominis ccrB1 into an MRSA strain carrying a type I SCCmec whose ccrB1 gene is inactive generated SCCmec excisants at a high frequency. The existence of an SCC without a mec determinant is indicative of a staphylococcal site-specific mobile genetic element that serves as a vehicle of transfer for various genetic markers between staphylococcal species.

  18. Dermatobia hominis: Small Migrants Hidden in Your Skin

    PubMed Central

    Zammarchi, Lorenzo; Viligiardi, Riccardo; Strohmeyer, Marianne

    2014-01-01

    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

  19. Vancomycin-resistant Staphylococcus hominis endophthalmitis following cataract surgery.

    PubMed

    Won, Jun Yeon; Kim, Moosang

    2013-01-01

    We report a case of acute postoperative endophthalmitis caused by vancomycin-resistant Staphylococcus hominis, treated at our hospital. An 80-year-old male presented 2 days after uncomplicated phacoemulsification and posterior chamber intraocular lens implantation, with a 24-hour history of progressive visual loss and redness in the operated (right) eye. On examination, best corrected visual acuity was counting fingers. Anterior segment examination revealed conjunctival injection, chemosis, corneal edema, and hypopyon. B-scan ultrasonography showed vitreous opacification, but no retinal detachment. Acute postoperative endophthalmitis was diagnosed. We performed vitrectomy with vancomycin in the irrigating solution, intraocular lens removal, and silicone oil tamponade. Culture of the vitreous grew Staphylococcus hominis. Antibiotic susceptibility testing showed the isolate was sensitive to trimethoprim/sulfamethoxazole and teicoplanin but resistant to ciprofloxacin, moxifloxacin, levofloxacin, cefazolin, and vancomycin. At 3 months, the visual acuity of the silicone oil-treated eye was 20/400.

  20. Sebaceous cysts with unpleasant twists: cutaneous myiasis with Dermatobia hominis.

    PubMed

    Osborne, M; O'Shearn, M K

    2013-01-01

    Dermatobia hominis (human Bot fly) causes furuncular myiasis (larval infection) in Central and South America. This report describes a case in a member of the UK Armed Forces who had recently taken part in an overseas training exercise in Belize. The importance of clinical history (including travel history) is highlighted. We also describe the outcomes of conservative treatment and surgical intervention for separate lesions in the same patient. PMID:24079201

  1. [Dermatobia hominis infection in a 3-year-old child].

    PubMed

    Meissner, M; Kippenberger, S; Valesky, E M; Kaufmann, R

    2012-04-01

    In the context of increasing travel to the tropics, outpatient services are more frequently confronted with non-domestic diseases in Europe. A 3-year old child presented with a painful tumor of the scalp. After incision of the furuncle-like lesion, we extracted a larva of the botfly Dermatobia hominis. Botflies are mainly encountered in Central and South America; they should be considered if patients demonstrate a furuncle-like lesion and have returned from a holiday in these endemic regions.

  2. Immunostimulation by phospholipopeptide biosurfactant from Staphylococcus hominis in Oreochromis mossambicus.

    PubMed

    Rajeswari, Veluchamy; Kalaivani Priyadarshini, Sekaran; Saranya, Viswanathan; Suguna, Ponnusamy; Shenbagarathai, Rajaiah

    2016-01-01

    The immunostimulatory effect of phospholipopeptide biosurfactant from Staphylococcus hominis (GenBank Accession No: KJ564272) was assessed with Oreochromis mossambicus. The non-specific (serum lysozyme activity, serum antiprotease activity, serum peroxidase activity and serum bactericidal activity), specific (bacterial agglutination assay) immune responses and disease resistance activity against Aeromonas hydrophila were examined. Fish were intraperitonially injected with water soluble secondary metabolite (biosurfactant) of S. hominis at a dose of 2 mg, 20 mg and 200 mg kg(-1) body weight. Commercial surfactant surfactin (sigma) at 20 mg kg(-1) was used as standard and saline as negative control. All the doses of water soluble biosurfactant tested, significantly enhanced the specific, nonspecific immunity and disease resistance from the day of post administration of phospholipopeptide biosurfactant till the tail of the experimental period. These results clearly indicated that the secondary metabolite isolated from S. hominis stimulates the immunity of finfish thereby could enhance aquaculture production.

  3. An emerging mycoplasma associated with trichomoniasis, vaginal infection and disease.

    PubMed

    Fettweis, Jennifer M; Serrano, Myrna G; Huang, Bernice; Brooks, J Paul; Glascock, Abigail L; Sheth, Nihar U; Strauss, Jerome F; Jefferson, Kimberly K; Buck, Gregory A

    2014-01-01

    Humans are colonized by thousands of bacterial species, but it is difficult to assess the metabolic and pathogenic potential of the majority of these because they have yet to be cultured. Here, we characterize an uncultivated vaginal mycoplasma tightly associated with trichomoniasis that was previously known by its 16S rRNA sequence as "Mnola." In this study, the mycoplasma was found almost exclusively in women infected with the sexually transmitted pathogen Trichomonas vaginalis, but rarely observed in women with no diagnosed disease. The genomes of four strains of this species were reconstructed using metagenome sequencing and assembly of DNA from four discrete mid-vaginal samples, one of which was obtained from a pregnant woman with trichomoniasis who delivered prematurely. These bacteria harbor several putative virulence factors and display unique metabolic strategies. Genes encoding proteins with high similarity to potential virulence factors include two collagenases, a hemolysin, an O-sialoglycoprotein endopeptidase and a feoB-type ferrous iron transport system. We propose the name "Candidatus Mycoplasma girerdii" for this potential new pathogen. PMID:25337710

  4. Recurrent abscesses due to Finegoldia magna, Dermabacter hominis and Staphylococcus aureus in an immunocompetent patient.

    PubMed

    Martin, J; Bemer, P; Touchais, S; Asseray, N; Corvec, S

    2009-10-01

    A case of recurrent abscesses in an immunocompetent patient is reported, involving the opportunistic human pathogen Dermabacter hominis, the virulent anaerobic pathogen Finegoldia magna and Staphylococcus aureus.

  5. 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

  6. Evaluation of amplified rDNA restriction analysis (ARDRA) for the identification of Mycoplasma species

    PubMed Central

    Stakenborg, Tim; Vicca, Jo; Butaye, Patrick; Maes, Dominiek; De Baere, Thierry; Verhelst, Rita; Peeters, Johan; de Kruif, Aart; Haesebrouck, Freddy; Vaneechoutte, Mario

    2005-01-01

    Background Mycoplasmas are present worldwide in a large number of animal hosts. Due to their small genome and parasitic lifestyle, Mycoplasma spp. require complex isolation media. Nevertheless, already over 100 different species have been identified and characterized and their number increases as more hosts are sampled. We studied the applicability of amplified rDNA restriction analysis (ARDRA) for the identification of all 116 acknowledged Mycoplasma species and subspecies. Methods Based upon available 16S rDNA sequences, we calculated and compared theoretical ARDRA profiles. To check the validity of these theoretically calculated profiles, we performed ARDRA on 60 strains of 27 different species and subspecies of the genus Mycoplasma. Results In silico digestion with the restriction endonuclease AluI (AG^CT) was found to be most discriminative and generated from 3 to 13 fragments depending on the Mycoplasma species. Although 73 Mycoplasma species could be differentiated using AluI, other species gave undistinguishable patterns. For these, an additional restriction digestion, typically with BfaI (C^TAG) or HpyF10VI (GCNNNNN^NNGC), was needed for a final identification. All in vitro obtained restriction profiles were in accordance with the calculated fragments based on only one 16S rDNA sequence, except for two isolates of M. columbinum and two isolates of the M. mycoides cluster, for which correct ARDRA profiles were only obtained if the sequences of both rrn operons were taken into account. Conclusion Theoretically, restriction digestion of the amplified rDNA was found to enable differentiation of all described Mycoplasma species and this could be confirmed by application of ARDRA on a total of 27 species and subspecies. PMID:15955250

  7. Genetic and Serological Analysis of Lipoprotein LppA in Mycoplasma mycoides subsp. mycoides LC and Mycoplasma mycoides subsp. capri

    PubMed Central

    Monnerat, Marie-Pierre; Thiaucourt, François; Poveda, Jose B.; Nicolet, Jacques; Frey, Joachim

    1999-01-01

    The genes encoding the 62-kDa lipoproteins from the Mycoplasma mycoides subsp. mycoides large-colony type (LC) strain Y-goat and the M. mycoides subsp. capri strain PG3 were cloned and analyzed by sequencing. These two lipoproteins have been named LppA[MmymyLC] and LppA[Mmyca], and their corresponding genes have been named lppA[MmymyLC] and lppA[Mmyca], respectively. The nucleotide and deduced amino acid sequences of these two lipoproteins showed a very high degree of similarity between these two mycoplasmas. Given the sequence data, LppA seems to fulfill the same structural functions as the previously described major lipoproteins P72 of M. mycoides subsp. mycoides small-colony type and P67 of the Mycoplasma species bovine group 7. Based on lppA gene sequences of M. mycoides subsp. mycoides LC and M. mycoides subsp. capri type strains, a specific PCR assay was developed so that it amplified this gene in all field strains of the two species analyzed in this study but not in the other members of the M. mycoides cluster. Analysis of the PCR-amplified lppA genes with frequently cutting restriction enzymes showed a certain degree of genetic variability which, however, did not cluster the two subspecies. This PCR therefore allows a rapid identification of M. mycoides subsp. mycoides LC and M. mycoides subsp. capri but does not distinguish between these two closely related subspecies. LppA was expressed in Escherichia coli K-12 and used for the production of polyclonal mouse antiserum. Antibodies against recombinant LppA[MmymyLC] reacted with a 62-kDa protein in all M. mycoides subsp. mycoides LC and M. mycoides subsp. capri type strains and field strains tested but not with the other members of the M. mycoides cluster, thus showing the antigenic specificity of LppA and further supporting the concept that a close relationship exists between these two mycoplasmas. PMID:10066658

  8. Antimicrobial susceptibility of Mycoplasma hyorhinis.

    PubMed

    Wu, C C; Shryock, T R; Lin, T L; Faderan, M; Veenhuizen, M F

    2000-09-15

    A broth microdilution technique was used to determine the antimicrobial susceptibility of 15 field isolates of Mycoplasma hyorhinis to 10 antimicrobial agents, representative of different classes, and contrasting newer agents to existing ones. For the macrolides, the MIC(90) for tylosin and tilmicosin was 1 and 4 microg/ml, respectively, but was > or = 16 microg/ml for erythromycin. Tetracycline, lincomycin and enrofloxacin each had an MIC(90) of 2 microg/ml. The mycoplasma had similar levels of susceptibility to the aminoglycoside and aminocyclictol classes exhibiting an MIC(90) of 4 microg/ml for gentamicin and 2 microg/ml for spectinomycin. The isolates exhibited high MICs to trimethoprim/sulfamethoxazole with an MIC(90) > or = 16/304 microg/ml. In summary, M. hyorhinis isolates from the US had low MICs against a variety of antimicrobials tested, with the exception of erythromycin and trimethoprim/sulfamethoxazole. PMID:10925038

  9. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 7 2013-04-01 2013-04-01 false Test for Mycoplasma. 610.30 Section 610.30 Food... GENERAL BIOLOGICAL PRODUCTS STANDARDS Mycoplasma § 610.30 Test for Mycoplasma. Except as provided... tested for the presence of Mycoplasma, as follows: Samples of the virus for this test shall be...

  10. 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 β(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 β(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

  11. Detection of specific Mycoplasma conjunctivae antibodies in the sera of sheep with infectious keratoconjunctivitis.

    PubMed

    Belloy, L; Giacometti, M; Abdo, E M; Nicolet, J; Krawinkler, M; Janovsky, M; Bruderer, U; Frey, J

    2001-01-01

    The serological cross reactions between Mycoplasma conjunctivae, the etiological agent of infectious keratoconjunctivitis (IKC), and the antigenetically and phylogenetically closely related Mycoplasma ovipneumoniae, which is often found in sheep, were analysed. Cross reacting antigens were identified using sera from sheep with IKC and from sheep of herds known to be free of IKC, as well as rabbit hyperimmune serum specific to the two Mycoplasma species. Cross reactions were predominantly due to the strongly antigenic proteins of 42 kDa and 83 kDa. Serospecific antigens of M. conjunctivae could be separated from cross-reacting antigens by the extraction of Tween 20-soluble membrane proteins. The Tween 20-extracted proteins of the M. conjunctivae strain HRC/581T were used for the development of an indirect ELISA test. This ELISA test was shown to be a useful serological method for the diagnosis of M. conjunctivae infections and to identify infected sheep herds.

  12. Mycoplasma neophronis sp. nov., isolated from the upper respiratory tract of Canarian Egyptian vultures (Neophron percnopterus majorensis).

    PubMed

    Suárez-Pérez, A; Ramírez, A S; Rosales, R S; Calabuig, P; Poveda, C; Rosselló-Móra, R; Nicholas, R A J; Poveda, J B

    2012-06-01

    Six strains with the typical characteristics of mycoplasmas were isolated from the tracheae of six Canarian Egyptian vultures (Neophron percnopterus majorensis). The results of biochemical, serological and molecular genetic studies showed that the isolates were nearly identical and that they could be considered as representing a novel species of the genus Mycoplasma. Colonies possessed the typical fried-egg appearance and electron micrographs revealed a pleomorphic cellular morphology with the lack of a cell wall. The isolates hydrolysed arginine and required sterol for growth but did not ferment glucose or hydrolyse urea. We propose that the isolates be assigned to a novel species,Mycoplasma neophronis sp. nov. The type strain is G.A.(T) ( = DSM 24097(T) = ATCC BAA-2157(T)). The antiserum of strain G.A.(T) has been deposited in the Mollicutes collection at Purdue University (Indiana, USA).

  13. Prevalence of Urogenital Mycoplasmas in Iran and Their Effects on Fertility Potential: A Systematic Review and Meta-Analysis

    PubMed Central

    AHMADI, Mohammad Hossein; MIRSALEHIAN, Akbar; BAHADOR, Abbas

    2016-01-01

    Background: Urogenital mycoplasmas are potentially pathogenic species causing genitourinary tract infections that may be initially asymptomatic but can progress and lead to severe complications and threaten reproductive health. However, the overall prevalence rate of this bacterium and its probable impacts on fertility potential have yet to be determined. Methods: We searched both English and Persian electronic databases using key words such as “Mycoplasma,” “Ureaplasma,” “M. hominis,” “M. genitalium,” “U. urealyticum,” “U. parvum,” “prevalence,” and “Iran”. Finally, after some exclusion, 29 studies from different regions of Iran were included in our study, and a meta-analysis was performed on collected data. Results: Urogenital mycoplasmas prevalence for women and men was high and ranged from 2%–40.5% and 2%–44.3%, respectively. The pooled prevalence in the male population was 11.1% (95% CI, 7.4%–16.4%) and in female was 12.8% (95% CI, 9.8%–16.5%). The prevalence of these bacteria was significantly higher in infertile men compared with that in fertile men. A high level of heterogeneity was observed for both men (I2 = 92.4%; P<0.001) and women (I2 = 93.3%; P<0.001). Some evidence for publication bias was observed in both men [Egger’s test (two-tailed P=0.0007), and Begg’s test (two-tailed P=0.0151)] and women [Egger’s test (two-tailed P=0.0006), and Begg’s test (two-tailed P=0.0086)] analysis. Conclusion: Since urogenital mycoplasmas may play a role in male infertility, screening strategies, particularly for asymptomatic individuals, and treatment of infected ones, which can reduce consequent complications, looks to be necessary. PMID:27252910

  14. IN VIVO AND IN VITRO EFFICACY OF NIGELLA SATIVA AQUEOUS EXTRACT ON BLASTOCYSTIS HOMINIS.

    PubMed

    Eida, Omima M; El-Shafei, Hanaa A; Nomeir, Yousra A; El Safhi, Mohammed B

    2016-04-01

    Metronidazole (MTZ) was the most widely accepted treatment for Blastocystis hominis (B. hominis) with high treatment failure rate, resistance and potential mutagenic and carcinogenic effects so there is urgent need to find out new, effective and safe treatment against B. hominis. The present research aimed to evaluate the therapeutic effect of the aqueous extract of Nigella sativa (NS) at different doses on B. hominis in vitro and in vivo in comparison to MTZ as a control drug. Isolates of B. hominis were obtained from patients complaining of diarrhea and abdominal pain. Isolates were cultured in egg diphasic medium (LE) for in vitro study and to adjust proper inoculating dose for in vivo study. The aqueous extract of NS at concentrations of 100 & 500 µg/ml showed a potent lethal effect on B. hominis isolates in vitro. Caecal tissue of experimentally infected and treated mice with two different doses of NS (250 & 500 mg/kg/d) were examined histopathologically and compared with that of mice infected and treated by two doses of MTZ (62 & 125 mg/kg/d) as control drug and Infected untreated mice as negative control group. Histopathological examination of infected untreated group showed all pathological degrees in the caecal tissue while infected treated one showed remission of pathological changes especially with higher dose (500 mg/kg). Present study proved that N. sativa had inhibitory effect on B. hominis in vitro and prevented cytopathic effect in infected mice inoculated orally with B. hominis. PMID:27363038

  15. Mycoplasma hyopneumoniae Transcription Unit Organization: Genome Survey and Prediction

    PubMed Central

    Siqueira, Franciele Maboni; Schrank, Augusto; Schrank, Irene Silveira

    2011-01-01

    Mycoplasma hyopneumoniae is associated with swine respiratory diseases. Although gene organization and regulation are well known in many prokaryotic organisms, knowledge on mycoplasma is limited. This study performed a comparative analysis of three strains of M. hyopneumoniae (7448, J and 232), with a focus on genome organization and gene comparison for open read frame (ORF) cluster (OC) identification. An in silico analysis of gene organization demonstrated 117 OCs and 34 single ORFs in M. hyopneumoniae 7448 and J, while 116 OCs and 36 single ORFs were identified in M. hyopneumoniae 232. Genomic comparison revealed high synteny and conservation of gene order between the OCs defined for 7448 and J strains as well as for 7448 and 232 strains. Twenty-one OCs were chosen and experimentally confirmed by reverse transcription–PCR from M. hyopneumoniae 7448 genome, validating our prediction. A subset of the ORFs within an OC could be independently transcribed due to the presence of internal promoters. Our results suggest that transcription occurs in ‘run-on’ from an upstream promoter in M. hyopneumoniae, thus forming large ORF clusters (from 2 to 29 ORFs in the same orientation) and indicating a complex transcriptional organization. PMID:22086999

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

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2014-01-01

    Different species of Mycoplasma can affect bovine cattle, causing several diseases. PCR sequencing and further analysis of the 16S-23S rRNA ITS region have shown a significant interspecies variability among Mollicutes. Sixteen suspected isolates of Mycoplasma spp. obtained from milk samples from dairy herds were amplified (16S-23S rRNA ITS region). Fourteen out of those 16 suspected Mycoplasma spp. isolates were PCR-positive. To confirm the identity of Mycoplasma bovis, these 14 isolates were tested by another species-specific PCR. Seven of the isolates rendered a positive result. The products of 16S-23S rRNA ITS PCR from one isolate that was identified as M. bovis and from two other isolates, identified as non- M. bovis were randomly selected, sequenced and analyzed. The three sequences (A, B and C) showed 100% similarity with M. bovis, Mycoplasma canadense and Mycoplasma californicum respectively. PMID:25011595

  18. Gliding Direction of Mycoplasma mobile

    PubMed Central

    Morio, Hanako; Kasai, Taishi

    2015-01-01

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

  19. Optimized PCR-based detection of mycoplasma.

    PubMed

    Dobrovolny, Paige L; Bess, Dan

    2011-06-20

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

  20. Efficacy of abamectin injection against Dermatobia hominis in cattle.

    PubMed

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

    1993-01-01

    The efficacy of abamectin 1%, when injected subcutaneously in cattle at a dose of 200 micrograms/kg body weight, against the larval stages (grubs) of the fly Dermatobia 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 on both sides of each animal before treatment, and were expressed, identified as to stage and classified as live or dead 10 days after treatment. Further larval counts were made periodically until day 79 to evaluate the degree of reinfestation and the stage of larval development. Reinfestation was first detected in the abamectin-treated cattle on day 44. Live larvae were found on 6-8 (Argentina) and on all (Brazil) controls at each post-treatment examination. The difference in numbers of live larvae between treatment groups was statistically significant (P < 0.05) at all post-treatment examinations. These data show that abamectin at a dose of 200 micrograms/kg body weight is highly effective in the treatment and control of established parasitic stages of D. hominis in cattle. No adverse reactions were observed in any of the treated animals. PMID:8493240

  1. Severe Mycoplasma bovis outbreak in an Austrian dairy herd.

    PubMed

    Pothmann, Harald; Spergser, Joachim; Elmer, Josef; Prunner, Isabella; Iwersen, Michael; Klein-Jöbstl, Daniela; Drillich, Marc

    2015-11-01

    A conventional dairy farm, housing 19 Austrian Simmental cows, experienced a spontaneous outbreak of a Mycoplasma bovis infection, showing severe clinical signs of respiratory tract disease, clinical mastitis, and tremendous drop in milk production. Despite intensive therapy, 5 cows died within 2 weeks or were euthanized. From the remaining cows, bacteriological culture and polymerase chain reaction revealed M. bovis in 10 of 14 milk samples. Mycoplasma bovis was found in 1 of 5 randomly collected nasal swabs. Autopsy of 1 cow revealed infection of the lungs and the udder with M. bovis. The 13 M. bovis isolates from milk samples, nasal swabs, lungs, and udder were genotyped by multilocus variable number of tandem-repeat analysis, and indicated that described infections were caused by a single M. bovis strain. The virulent M. bovis strain resulted in dramatic economic loss to the farmer. To control the disease, culling of all animals, including heifers and calves, was recommended, and strict hygienic measures were implemented before introducing new animals to the farm. PMID:26450838

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

    PubMed Central

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

    1996-01-01

    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

  3. Antimicrobial susceptibility and molecular characterization of macrolide resistance of Mycoplasma bovis isolates from multiple provinces in China

    PubMed Central

    KONG, Ling-Cong; GAO, Duo; JIA, Bo-Yan; WANG, Zi; GAO, Yun-Hang; PEI, Zhi-Hua; LIU, Shu-Ming; XIN, Jiu-Qing; MA, Hong-Xia

    2015-01-01

    Mycoplasma bovis has spread widely throughout the world via animal movement and has become an important pathogen of bovine respiratory disease. However, the minimum inhibitory concentrations of antimicrobials for Mycoplasma bovis have not been studied in China. The objective of this study was to determine the prevalence and antibiotic resistance of Mycoplasma bovis isolated from young cattle with respiratory infection in China. Mycoplasma bovis was detected in 32/45 bovine respiratory infection outbreaks at beef farms in 8 provinces in China. The isolates were susceptible or had medium sensitivity to ciprofloxacin, enrofloxacin and doxycycline, but were frequently resistant to macrolides (13/32, 41%). An A2058G (Escherichia coli Numbering) mutation located in the rrnA operon in domain V of 23S rRNA was observed in strains that were resistant to macrolides. This single mutations at the rrnA operon in domain V of 23S rRNA may play an important role in the resistance of Mycoplasma bovis strains to macrolides. PMID:26346744

  4. Population identification of Sarcoptes hominis and Sarcoptes canis in China using DNA sequences.

    PubMed

    Zhao, YaE; Cao, ZhiGuo; Cheng, Juan; Hu, Li; Ma, JunXian; Yang, YuanJun; Wang, XiaoPeng; Zeng, JiHui; Wang, TianPing

    2015-03-01

    There has been no consistent conclusion on whether Sarcoptes mites parasitizing in humans and animals are the same species. To identify Sarcoptes (S.) hominis and S. canis in China, gDNA was extracted from individual mites (five from patients with scabies and five from dogs with mange) for amplification of rDNA ITS2, mtDNA 16S, and cox1 fragment sequences. Then, the sequences obtained were aligned with those from different hosts and geographical locations retrieved from GenBank and sequence analyses were conducted. Phylogenetic trees based on 317-bp mtDNA cox1 showed five distinctive branches (species) of Sarcoptes mites, four for S. hominis (S. hominis Chinese, S. nr. hominis Chinese, S. hominis Australian, and S. hominis Panamanian) and one for S. animal (S. animal). S. animal included mites from nine animal species, with S. canis China, S. canis Australia, and S. canis USA clustering as a subbranch. Further sequence divergence analysis revealed no overlap between intraspecific (≤ 2.6 %) and interspecific (2.6-10.5 %) divergences in 317-bp mtDNA cox1. However, overlap was detected between intra- and interspecific divergences in 311-bp rDNA ITS2 or 275-bp mtDNA 16S when the divergences exceeded 1.0 %, which resulted in failure in identification of Sarcoptes. The results showed that the 317-bp mtDNA cox1 could be used as a DNA barcode for molecular identification of Sarcoptes mites. In addition, geographical isolation was observed between S. hominis Chinese, S. hominis Australian, and S. hominis Panamanian, but not between all S. canis. S. canis and the other S. animal belonged to the same species. PMID:25547078

  5. Mycoplasmas and Ureaplasmas as Neonatal Pathogens

    PubMed Central

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

    2005-01-01

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

  6. Seroepidemiological survey of sheep flocks from Northern Japan for Mycoplasma ovipneumoniae and Mycoplasma agalactiae.

    PubMed

    Giangaspero, Massimo; Nicholas, Robin A J; Hlusek, Miroslav; Bonfini, Barbara; Osawa, Takeshi; Orusa, Riccardo; Tatami, Shingo; Takagi, Eishu; Moriya, Hiroaki; Okura, Norimoto; Kato, Kazuo; Kimura, Atsushi; Harasawa, Ryô; Ayling, Roger D

    2012-03-01

    Sheep flocks from Hokkaido, Iwate and Aomori, three northern prefectures of Japan, were screened for antibodies to Mycoplasma ovipneumoniae and Mycoplasma agalactiae by ELISA. Sixty four animals out of 246 (26%) were seropositive to M. ovipneumoniae, with positive results obtained from all three prefectures. None of the sera tested were serologically positive to M. agalactiae.

  7. In vitro antimicrobial susceptibility of Mycoplasma bovis isolated in Israel from local and imported cattle.

    PubMed

    Gerchman, Irena; Levisohn, Sharon; Mikula, Inna; Lysnyansky, Inna

    2009-06-12

    Monitoring of susceptibility to antibiotics in field isolates of pathogenic bovine mycoplasmas is important for appropriate choice of treatment. Our study compared in vitro susceptibility profiles of Mycoplasma bovis clinical strains, isolated during 2005-2007 from Israeli and imported calves. Minimal inhibitory concentration (MIC) values were determined for macrolides by the microbroth dilution test, for aminoglycosides by commercial Etest, and for fluoroquinolones and tetracyclines by both methods. Notably, although correlation between the methods was generally good, it was not possible to determine the MIC endpoint for enrofloxacin-resistant strains (MIC > or =2.5 microg/ml in the microtest) by Etest. Comparison of antibiotic susceptibility profiles between local and imported M. bovis strains revealed that local strains were significantly more resistant to macrolides than most isolates from imported animals, with MIC(50) of 128 microg/ml vs. 2 microg/ml for tilmicosin and 8 microg/ml vs. 1 microg/ml for tylosin, respectively. However, local strains were more susceptible than most imported strains to fluoroquinolones and spectinomycin. Difference in susceptibility to tetracycline, doxycycline and oxytetracycline between local and imported strains was expressed in MIC(90) values for imported strains in the susceptible range compared to intermediate susceptibility for local strains. The marked difference in susceptibility profiles of M. bovis strains isolated from different geographical regions seen in this study emphasizes the necessity for performing of the antimicrobial susceptibility testing periodically and on a regional basis. PMID:19250777

  8. Histopathological findings, phenotyping of inflammatory cells, and expression of markers of nitritative injury in joint tissue samples from calves after vaccination and intraarticular challenge with Mycoplasma bovis strain 1067

    PubMed Central

    2014-01-01

    Background The pathogenesis of caseonecrotic lesions developing in lungs and joints of calves infected with Mycoplasma bovis is not clear and attempts to prevent M. bovis-induced disease by vaccines have been largely unsuccessful. In this investigation, joint samples from 4 calves, i.e. 2 vaccinated and 2 non-vaccinated, of a vaccination experiment with intraarticular challenge were examined. The aim was to characterize the histopathological findings, the phenotypes of inflammatory cells, the expression of class II major histocompatibility complex (MHC class II) molecules, and the expression of markers for nitritative stress, i.e. inducible nitric oxide synthase (iNOS) and nitrotyrosine (NT), in synovial membrane samples from these calves. Furthermore, the samples were examined for M. bovis antigens including variable surface protein (Vsp) antigens and M. bovis organisms by cultivation techniques. Results The inoculated joints of all 4 calves had caseonecrotic and inflammatory lesions. Necrotic foci were demarcated by phagocytic cells, i.e. macrophages and neutrophilic granulocytes, and by T and B lymphocytes. The presence of M. bovis antigens in necrotic tissue lesions was associated with expression of iNOS and NT by macrophages. Only single macrophages demarcating the necrotic foci were positive for MHC class II. Microbiological results revealed that M. bovis had spread to approximately 27% of the non-inoculated joints. Differences in extent or severity between the lesions in samples from vaccinated and non-vaccinated animals were not seen. Conclusions The results suggest that nitritative injury, as in pneumonic lung tissue of M. bovis-infected calves, is involved in the development of caseonecrotic joint lesions. Only single macrophages were positive for MHC class II indicating down-regulation of antigen-presenting mechanisms possibly caused by local production of iNOS and NO by infiltrating macrophages. PMID:25162202

  9. Genome Sequence of Mycoplasma hyorhinis Isolated from Cell Cultures

    PubMed Central

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

    2016-01-01

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

  10. Mycoplasmas hyorhinis in different regions of cuba. diagnosis

    PubMed Central

    Lobo, Evelyn; Poveda, Carlos; Gupta, Rakesh; Suarez, Alejandro; Hernández, Yenney; Ramírez, Ana; Poveda, José B.

    2011-01-01

    M. hyorhinis is considered one of the etiological agents of arthritis in sucking pigs, but recently as seen, some strains can produce pneumonia that could not be distinguished from the mycoplasmosis caused by M. hyopneumoniae. The study was conducted to research the presence of Mycoplasma hyorhinis (M. hyorhinis ) in different regions of the country from exudates of pig lungs with typical EP lesions. Exudates from 280 pig lungs with typical EP lesions were studied using molecular techniques such as PCR, real time PCR and amplification of the 16S-23S rRNA. It was detected that the 66% of the samples studied resulted positive to M. hyorhinis, and the presence of this species was detected in all the provinces. Amplification and studies on the intergenic region 16S-23S of M. hyorhinis rRNA demonstrated the existing variability among strains of a same species. This study is the first report on M. hyorhinis detection in Cuba. PMID:24031686

  11. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Test for Mycoplasma. 610.30 Section 610.30 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS GENERAL BIOLOGICAL PRODUCTS STANDARDS Mycoplasma § 610.30 Test for Mycoplasma. Except as provided otherwise in this subchapter, prior to...

  12. The experimental infection of specific pathogen free lambs with Mycoplasma ovipneumoniae.

    PubMed

    Foggie, A; Jones, G E; Buxton, D

    1976-07-01

    Six colostrum-deprived SPF lambs inoculated endobronchially with a second passage broth culture of a Scottish strain of Mycoplasma ovipneumoniae, were killed in batches of two at seven, 14 and 28 days post-inoculation. One lamb from each batch showed macroscopic and microscopic lung lesions similar to but milder than those described for respiratory mycoplasmoses in other species of animals and exhibited minor clinical symptoms. Mycoplasma were recovered from all infected but from no control animals: five infected lambs yielded mycoplasma from lung tissue. Two lambs infected with M ovipneumoniae by endobronchial intubation were placed in contact with six other SPF lambs. M ovipneumoniae was recovered from the upper respiratory tract only of all six contact lambs, but no pathological changes were noted in their lungs. Both donor lambs yielded mycoplasma from lung tissue, but microscopic lesions were detected in only one of them, and these were minimal. No seroconversion due to the infection could be demonstrated in any of the lambs by either the indirect haemagglutination or metabolic inhibition tests.

  13. Molecular characterization of the heat shock protein 70 gene in Mycoplasma ovipneumoniae.

    PubMed

    Zhang, Bin; Han, Xiao; Yue, Hua; Tang, Cheng

    2013-10-01

    Mycoplasma ovipneumoniae is a species of mycoplasma bacteria that commonly infects the respiratory tract, causing respiratory disease in sheep and goats worldwide. In the current study, the 70-kDa heat shock protein (Hsp70) gene was cloned, sequenced and analyzed in 14 clinical isolates of M. ovipneumoniae. Results showed that, compared to the reference Y98 strain, the open-reading frames (ORFs) of Hsp70 gene in all isolates were 1818 base pairs (bp). Three nucleotides of TCA were inserted at 1,776 bp, resulting in insertion of the amino acid glutamine at amino acid position 593. The neighbor-joining trees, constructed using the Hsp70 gene, exhibited that the closest genetic relationship occurred between M. ovipneumoniae and Mycoplasma hyopneumoniae, which was consistent with the one based on the whole genome comparisons between these two mycoplasma species. Therefore, these results suggest that the Hsp70 gene, rather than 16S ribosomal RNA, was suitable as a potential molecular marker for evaluating the genetic relationship of M. ovipneumoniae with other bacterial species.

  14. In Vitro Spatial and Temporal Analysis of Mycoplasma pneumoniae Colonization of Human Airway Epithelium

    PubMed Central

    Prince, Oliver A.; Krunkosky, Thomas M.

    2014-01-01

    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

  15. Multidrug-resistant Staphylococcus hominis subsp. novobiosepticus causing septicemia in patients with malignancy.

    PubMed

    Roy, Priyamvada; Ahmed, Nishat Hussain; Biswal, Indu; Grover, Rajesh Kumar

    2014-01-01

    A new subspecies of Staphylococcus hominis described by Kloos et al. in 1998 and named S. hominis subsp. novobiosepticus (SHN) has been implicated in nosocomial outbreaks. Multidrug resistance, including resistance to novobiocin and oxacillin, is a particularly important feature of SHN. In our institute, we encountered 13 cases of S. hominis subsp. hominis in cancer patients with septicemia, of which seven were methicillin resistant. The isolates were identified by VITEK ® 2 compact automated system, using GP REF 21342 identification card and antimicrobial susceptibility testing card P-628. The biochemical reactions and antibiotic susceptibility pattern of the seven methicillin-resistant isolates were re-analyzed and patient details were re-checked to finally identify them as SHN. The increasing number of cases reporting isolation of SHN from biological specimens point to potential virulence and clinical importance of this bacterium.

  16. Recurrent abscesses due to Finegoldia magna, Dermabacter hominis and Staphylococcus aureus in an immunocompetent patient.

    PubMed

    Martin, J; Bemer, P; Touchais, S; Asseray, N; Corvec, S

    2009-10-01

    A case of recurrent abscesses in an immunocompetent patient is reported, involving the opportunistic human pathogen Dermabacter hominis, the virulent anaerobic pathogen Finegoldia magna and Staphylococcus aureus. PMID:19332143

  17. Experimental evaluation of inactivated and live attenuated vaccines against Mycoplasma mycoides subsp. mycoides.

    PubMed

    Mwirigi, Martin; Nkando, Isabel; Aye, Racheal; Soi, Reuben; Ochanda, Horace; Berberov, Emil; Potter, Andrew; Gerdts, Volker; Perez-Casal, Jose; Naessens, Jan; Wesonga, Hezron

    2016-01-01

    The current control method for contagious bovine pleuropneumonia (CBPP) in Africa is vaccination with a live, attenuated strain of Mycoplasma mycoides subsp. mycoides (Mmm). However, this method is not very efficient and often causes serious adverse reactions. Several studies have attempted to induce protection using inactivated mycoplasma, but with widely contradictory results. Therefore, we compared the protective capacity of the live T1/44 vaccine with two inactivated preparations of Mmm strain Afadé, inoculated with an adjuvant. Protection was measured after a challenge with Afadé. The protection levels were 31%, 80.8% and 74.1% for the formalin-inactivated, heat-inactivated and live attenuated preparations, respectively. These findings indicate that low doses of heat-inactivated Mmm can offer protection to a level similar to the current live attenuated (T1/44) vaccine formulation.

  18. Molecular demonstration of hemotropic mycoplasmas in wild Japanese monkeys (Macaca fuscata).

    PubMed

    Sashida, Hinako; Suzuki, Yoshihisa; Rokuhara, Sou; Nagai, Kazuya; Harasawa, Ryô

    2014-01-01

    The prevalence of hemotropic mycoplasmas in wild monkeys is largely unknown. Here, we report the presence of hemoplasmas in blood specimens collected from wild Japanese monkeys (Macaca fuscata) tentatively captured for ecological survey in Mie prefecture, Japan. We examined 9 monkeys using hemoplasma-specific real-time PCR and found all of them positive for a hemoplasma infection. The 16S rRNA gene and 16S to 23S rRNA intergenic spacer region of the hemoplasma detected in wild monkeys were amplified using end-point PCR. The nucleotide sequences of the PCR products were further determined and compared to those of other hemoplasmas. Our examinations revealed a wide prevalence of a hemoplasma strain in Japanese monkeys, which was similar to 'Candidatus Mycoplasma haemomacaque' reported in cynomolgus monkeys (Macaca fascicularis). Pathogenic traits of this hemoplasma strain remain unexplored.

  19. Experimental evaluation of inactivated and live attenuated vaccines against Mycoplasma mycoides subsp. mycoides.

    PubMed

    Mwirigi, Martin; Nkando, Isabel; Aye, Racheal; Soi, Reuben; Ochanda, Horace; Berberov, Emil; Potter, Andrew; Gerdts, Volker; Perez-Casal, Jose; Naessens, Jan; Wesonga, Hezron

    2016-01-01

    The current control method for contagious bovine pleuropneumonia (CBPP) in Africa is vaccination with a live, attenuated strain of Mycoplasma mycoides subsp. mycoides (Mmm). However, this method is not very efficient and often causes serious adverse reactions. Several studies have attempted to induce protection using inactivated mycoplasma, but with widely contradictory results. Therefore, we compared the protective capacity of the live T1/44 vaccine with two inactivated preparations of Mmm strain Afadé, inoculated with an adjuvant. Protection was measured after a challenge with Afadé. The protection levels were 31%, 80.8% and 74.1% for the formalin-inactivated, heat-inactivated and live attenuated preparations, respectively. These findings indicate that low doses of heat-inactivated Mmm can offer protection to a level similar to the current live attenuated (T1/44) vaccine formulation. PMID:26827840

  20. 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

  1. 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.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  3. Eosinophilic Fasciitis Associated with Mycoplasma arginini Infection

    PubMed Central

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

    2012-01-01

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

  4. In vitro susceptibility of avian mycoplasmas to enrofloxacin, sarafloxacin, tylosin, and oxytetracycline.

    PubMed

    Wang, C; Ewing, M; Aarabi, S Y

    2001-01-01

    In vitro susceptibility of avian Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) to enrofloxacin, sarafloxacin, tylosin, and oxytetracycline was determined by a serial broth dilution method. The minimum inhibitory concentration (MIC) was recognized by a conversion of the pH indicator phenol red in culture media to a yellow color. Each isolate or type strain of mycoplasma was tested in two replicates. The MICs of tylosin, enrofloxacin, sarafloxacin, and oxytetracycline against five isolates and two reference strains of MG (approximately 10(5) colony-forming units [CFU]/ml) were 0.05, 0.14, 0.37, and 1.30 microg/ml, respectively. The MICs of the four antimicrobial agents against six isolates and one reference strain of MS (approximate 10(5) CFU/ml) were 0.13, 1.82, 1.76, and 0.91 microg/ml, respectively. There were no differences (P > 0.05) between tylosin, enrofloxacin, and sarafloxacin against MG, but these three antibiotics were different (P < 0.05) from oxytetracycline. The MIC value of tylosin against MS was different (P < 0.05) from those of sarafloxacin and enrofloxacin, but it was not different (P > 0.05) from that of oxytetracycline. PMID:11417828

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

    PubMed Central

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

    1984-01-01

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

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

    USGS Publications Warehouse

    Goldberg, D.R.; Samuel, M.D.; Thomas, C.B.; Sharp, P.; Krapu, G.L.; Robb, J.R.; Kenow, K.P.; Korschgen, C.E.; Chipley, W.H.; Conroy, M.J.; Kleven, S.H.

    1995-01-01

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

  7. Antigenic heterogeneity in Mycoplasma iowae demonstrated with monoclonal antibodies.

    PubMed

    Panangala, V S; Gresham, M M; Morsy, M A

    1992-01-01

    Western blots of proteins of 14 Mycoplasma iowae strains and isolates resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis were probed with three monoclonal antibodies (MAbs), MI6, MI7, and MI8. MAb MI6 reacted with one or more antigens with apparent molecular weights of 60,000, 70,000, and 94,000. In three strains (N-PHN-D13, R-D2497, and K 1805), antigens located on a single peptide band were recognized, while in others additional epitopes at different molecular-weight positions were revealed. A similar pattern was observed with MAb MI7, although it reacted with fewer antigens than did MAb MI6 and failed to recognize antigens in strains N-PHN-D13 and R-D2497. MAb MI8 reacted with an antigen at an apparent molecular-weight position of 28,000 in four of the 14 strains and isolates. The diverse reaction patterns observed with the MAbs in the 14 M. iowae strains and isolates confirms the occurrence of antigenic variation within this species. Antigenic variation in M. iowae may be pivotal in determining host-parasite interactions, pathogenesis, and the outcome of disease. PMID:1373600

  8. Mycoplasma gallopavonis in eastern wild turkeys.

    PubMed

    Luttrell, M P; Eleazer, T H; Kleven, S H

    1992-04-01

    Serum samples and tracheal cultures were collected from eastern wild turkeys (Meleagris gallopavo sylvestris) trapped for relocation in South Carolina (USA) during 1985 to 1990. Sera were tested for Mycoplasma gallisepticum and M. synoviae by the rapid plate agglutination and hemagglutination inhibition tests and were found to be negative. Tracheal cultures were negative for all pathogenic Mycoplasma spp., including M. gallisepticum, M. synoviae, M. meleagridis, and M. iowae. However, M. gallopavonis was isolated from every group of wild turkeys tested in 1986 to 1990. These data suggest that M. gallopavonis, which is generally considered nonpathogenic, may be a common microorganism in eastern wild turkeys.

  9. Cryptosporidium hominis gene catalog: a resource for the selection of novel Cryptosporidium vaccine candidates

    PubMed Central

    Ifeonu, Olukemi O.; Simon, Raphael; Tennant, Sharon M.; Sheoran, Abhineet S.; Daly, Maria C.; Felix, Victor; Kissinger, Jessica C.; Widmer, Giovanni; Levine, Myron M.; Tzipori, Saul; Silva, Joana C.

    2016-01-01

    Human cryptosporidiosis, caused primarily by Cryptosporidium hominis and a subset of Cryptosporidium parvum, is a major cause of moderate-to-severe diarrhea in children under 5 years of age in developing countries and can lead to nutritional stunting and death. Cryptosporidiosis is particularly severe and potentially lethal in immunocompromised hosts. Biological and technical challenges have impeded traditional vaccinology approaches to identify novel targets for the development of vaccines against C. hominis, the predominant species associated with human disease. We deemed that the existence of genomic resources for multiple species in the genus, including a much-improved genome assembly and annotation for C. hominis, makes a reverse vaccinology approach feasible. To this end, we sought to generate a searchable online resource, termed C. hominis gene catalog, which registers all C. hominis genes and their properties relevant for the identification and prioritization of candidate vaccine antigens, including physical attributes, properties related to antigenic potential and expression data. Using bioinformatic approaches, we identified ∼400 C. hominis genes containing properties typical of surface-exposed antigens, such as predicted glycosylphosphatidylinositol (GPI)-anchor motifs, multiple transmembrane motifs and/or signal peptides targeting the encoded protein to the secretory pathway. This set can be narrowed further, e.g. by focusing on potential GPI-anchored proteins lacking homologs in the human genome, but with homologs in the other Cryptosporidium species for which genomic data are available, and with low amino acid polymorphism. Additional selection criteria related to recombinant expression and purification include minimizing predicted post-translation modifications and potential disulfide bonds. Forty proteins satisfying these criteria were selected from 3745 proteins in the updated C. hominis annotation. The immunogenic potential of a few of these is

  10. 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

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

    PubMed

    Muñoz, G; Sotomayor, P

    1990-07-01

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

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

    PubMed

    Muñoz, G; Sotomayor, P

    1990-07-01

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

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

    PubMed Central

    Muñoz, G; Sotomayor, P

    1990-01-01

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

  14. Prevalence and characteristics of Blastocystis hominis infection in children.

    PubMed

    O'Gorman, M A; Orenstein, S R; Proujansky, R; Wadowsky, R M; Putnam, P E; Kocoshis, S A

    1993-02-01

    Blastocystis hominis, a protozoan whose pathogenicity has been questioned, is sometimes found in the human gastrointestinal tract. We sought to determine the prevalence of Blastocystis in stool and to characterize clinical features of infection with Blastocystis in children. Forty-six (3%) of 1,736 patients undergoing fecal microscopy at Children's Hospital of Pittsburgh between January 1, 1985, and December 31, 1988, harbored Blastocystis. Of these 46 children, 75% had exposure to well water or had been in developing countries. Thirty-nine of the 46 (85%) experienced gastrointestinal symptoms, such as abdominal pain, diarrhea, vomiting, and weight loss. Blastocystis was the only parasite found in 35 of those 39 symptomatic children. Symptoms resolved within one month in 90% of patients receiving antiparasitic pharmacotherapy, but in only 58% (P < .04) of those receiving no therapy. We conclude that children infected with Blastocystis often experience gastrointestinal symptoms and that treatment increases the rate of symptomatic improvement. We speculate that Blastocystis is a human pathogen. PMID:8432086

  15. Isolation and molecular identification of Mycoplasma equigenitalium from equine genital tracts in northern India

    PubMed Central

    Nehra, K; Rana, R; Viswas, K. N; Arun, T. R.; Singh, V. P.; Singh, A. P; Prabhu, S. N

    2015-01-01

    Although Mycoplasma equigenitalium has been implicated in equine reproductive problems, its prevalence is largely unexplored due to the lack of specific diagnostic tests. To address this limitation, the authors developed and optimized species-specific primer pairs that target M. eguigenitalium rpoB (RNA polymerase B subunit) gene sequences. The specificity of the PCR assay developed in this study was determined using 12 field isolates including the type strain of M. equigenitalium and other Mycoplasma species. In the field study, a total of 122 mare and stallion samples comprising of 50 clinical and 72 random samples were subjected to species-specific PCR assay to detect M. equigenitalium in equine genital tracts. Mycoplasma equigenitalium (MEG) species-specific PCR detected 22.13% positive samples; however, only 9.01% of the samples were found to be positive using the conventional culture technique. The PCR established in this study could be used for rapid, specific and accurate diagnosis of M. equigenitalium strains. To the authors’ knowledge, this is the first report addressing the development and evaluation of species-specific PCR to detect M. equigenitalium. PMID:27175172

  16. Characterization of the in vitro core surface proteome of Mycoplasma mycoides subsp. mycoides, the causative agent of contagious bovine pleuropneumonia.

    PubMed

    Krasteva, Ivanka; Liljander, Anne; Fischer, Anne; Smith, David G E; Inglis, Neil F; Scacchia, Massimo; Pini, Attilio; Jores, Joerg; Sacchini, Flavio

    2014-01-10

    Contagious bovine pleuropneumonia (CBPP), caused by Mycoplasma mycoides subsp. mycoides (Mmm) is a severe cattle disease, present in many countries in sub-Saharan Africa. The development of improved diagnostic tests and vaccines for CBPP control remains a research priority. Polyacrylamide gel electrophoresis and mass spectrometry were used to characterize the Triton X-114 soluble proteome of nine Mmm strains isolated from Europe or Africa. Of a total of 250 proteins detected, 67 were present in all strains investigated. Of these, 44 were predicted to be lipoproteins or cytoplasmic membrane-associated proteins and are thus likely to be members of the core in vitro surface membrane-associated proteome of Mmm. Moreover, the presence of all identified proteins in other ruminant Mycoplasma pathogens were investigated. Two proteins of the core proteome were identified only in other cattle pathogens of the genus Mycoplasma pointing towards a role in host-pathogen interactions. The data generated will facilitate the identification and prioritization of candidate Mycoplasma antigens for improved control measures, as it is likely that surface-exposed membrane proteins will include those that are involved in host-pathogen interactions.

  17. The prevalence of Blastocystis hominis and other protozoan parasites in soldiers returning from peacekeeping missions.

    PubMed

    Duda, Aleksandra; Kosik-Bogacka, Danuta; Lanocha-Arendarczyk, Natalia; Kołodziejczyk, Lidia; Lanocha, Aleksandra

    2015-04-01

    Blastocystis hominis is a common intestinal parasite found in humans living in poor sanitary conditions, living in tropical and subtropical climates, exposed to infected animals, or consuming contaminated food or water. The aim of this study was to determine the prevalence of B. hominis in Polish military personnel returning from peacekeeping missions in Iraq and Afghanistan. In total, 1,826 stool samples were examined. Gastrointestinal parasites were detected in 17% of the soldiers. The examined stool samples most frequently contained vacuolar forms of B. hominis (15.3%) and cysts of Entamoeba coli (1.0%) or Giardia lamblia (0.7%). In 97.1% of stool samples from infected soldiers, we observed less than five developmental forms of B. hominis in the field of view (40×). The parasite infections in soldiers were diagnosed in the autumn and the spring. There was no statistical correlation between age and B. hominis infection. Our results show that peacekeeping missions in countries with tropical or subtropical climates could be associated with risk for parasitic diseases, including blastocystosis.

  18. Blastocystis hominis: phylogenetic affinities determined by rRNA sequence comparison.

    PubMed

    Johnson, A M; Thanou, A; Boreham, P F; Baverstock, P R

    1989-04-01

    In 1912 Blastocystis hominis was identified as a new species and classified as a yeast (Brumpt 1912). In the early 1920s several groups confirmed its classification as a yeast, specifically a member of the genus Schizosaccharomyces (discussed by Zierdt et al. 1967). Apart from an occasional case report, the classification of B. hominis and its role as a harmless intestinal yeast was not questioned for another 50 years. Then, Zierdt (1967) suggested that it should be classified in the phylum Protozoa, subphylum Sporozoa, and that it should be considered as a potential pathogen. The likely role of B. hominis as a human pathogen has recently become more firmly established (Garcia et al. 1984; Sheehan et al. 1986) and its classification has been changed. Although the classification of B. hominis as a protozoon was assumed widely, classification as a sporozoon was not accepted, and the most recent definitive classification of the Protozoa did not even list B. hominis (Lee et al. 1985). Then, based essentially on a review of the known characteristics of the organism, it was recently reclassified into the subphylum Sarcodina (Zierdt 1988). Clearly, the phylogeny of this emerging human pathogen needs definitive analysis (Mehlhorn 1988). PMID:2649390

  19. Clinical characteristics and endoscopic findings associated with Blastocystis hominis in healthy adults.

    PubMed

    Chen, Te-Li; Chan, Che-Chang; Chen, Hsin-Pai; Fung, Chang-Phone; Lin, Chih-Pei; Chan, Wan-Leong; Liu, Cheng-Yi

    2003-08-01

    Ninety-nine individuals with stools positive for Blastocystis hominis but negative for other parasites were identified from medical records of healthy adults who had received a physical examination at Taipei Veterans General Hospital from November 2000 to October 2002. The medical records of these 99 positive cases and 193 randomly selected controls, matched for age, sex, and date of examination, were retrospectively reviewed. The pathogenicity of B. hominis could not be demonstrated due to a lack of association with the development of gastrointestinal symptoms or pathologic findings on endoscopic examination. Multivariate analyses revealed that chronic hepatitis B infection was a predisposing condition to the acquisition of B. hominis (odd ratio = 2.848, 95% confidence interval = 1.299-6.242, P = 0.009), and concentration of urate was significantly lower in B. hominis-positive individuals (mean +/- SD = 361.64 +/- 87.44 versus 392.57 +/- 93.38 micromol/L; P = 0.009). Among the 64 individuals who underwent gastric biopsy, Helicobacter pylori was found more frequently in the individuals harboring B. hominis (19 of 26 versus 15 of 38; P = 0.017). PMID:13677378

  20. [Blastocystis hominis in patients at the Ruiz y Paez University Hospital from Bolivar City, Venezuela].

    PubMed

    Devera, R; Azacon, B; Jiménez, M

    1998-01-01

    Blastocystis hominis is a polymorphic protozoan of discussed taxonomic position, which is currently associated with human intestinal disease. In order to determine the prevalence of the microorganism in a sample of hospitalized patients, a study was carried out from november 1996 to april 1997 on 100 adult patients of both sexes aged 20 to 79 years at the "Ruíz y Páez" University Hospital of Bolivar city, Venezuela. A coproparasitological study was carried out using direct examination and Faust method. Infection by parasites and/or commensals was demonstrated in 48 patients. The most frequent agent was B. hominis with a prevalence of 42.0%. We did not find a statistically association between sex (P > 0.05) or age (X2 = 3.52; d.f; = 3) and B. hominis infection. B. hominis was most frequently identified as the single parasite (88.1%), and with a number of less than 5 cells per 400X microscopic field (73.8%). The infection was more common in patients with base chronic-immunosuppressive diseases, the major one being cancer. Diarrhea was observed in 27.0% of cases. Due to its high prevalence, especially as a single agent, together with the particular immunological characteristics of the patients studied, a potential pathogenic role of the opportunistic type is suggested for B. hominis. PMID:10413881

  1. Pentatrichomonas hominis: prevalence and molecular characterization in humans, dogs, and monkeys in Northern China.

    PubMed

    Li, Wen-Chao; Ying, Meng; Gong, Peng-Tao; Li, Jian-Hua; Yang, Ju; Li, He; Zhang, Xi-Chen

    2016-02-01

    Pentatrichomonas hominis is an anaerobic amitochondrial flagellated protist that primarily colonizes the large intestines of a number of species, including cats, dogs, nonhuman primates, and humans. The prevalence of this parasite in dogs, monkeys, and humans is, however, poorly understood. In this study, a total of 362 fecal samples including 252 dogs, 60 monkeys, and 50 humans from northern China were collected for an epidemiological survey of P. hominis infection.The average prevalence of P. hominis infection determined by nested PCR was 27.38% (69/252), 4.00% (2/50), and 46.67% (28/60) in dogs, humans, and monkeys, respectively. The prevalence was significantly higher in 6-month-old dogs (41.53%) and children (7.69%) than in older dogs (14.39%) and adults (0%) (P < 0.05). Sequencing of amplicons revealed that four variable positions separated sequences into three types, called CC1-3. CC1 was the most prevalent in the study population. This study determined that P. hominis infection is common in dogs, monkeys, and humans, especially in children and young dogs. Given the infection prevalence, P. hominis may pose a risk of zoonotic and anthroponotic transmission.

  2. [Prevalence of Blastocystis hominis among food handlers from Caroni municipality, Bolivar State, Venezuela].

    PubMed

    Requena, Ixora; Hernández, Yessica; Ramsay, Mario; Salazar, Carmen; Devera, Rodolfo

    2003-01-01

    A cross-sectional survey was conducted to determine the prevalence of Blastocystis hominis infection in a random sample of apparently healthy food handlers. A total of 415 individuals attending the Manoa Urban Outpatient Clinic (Caroní Municipality, Bolívar State, Venezuela) in the Adult Hygiene Program and who requested health certification to work as food handlers were studied. Stool samples obtained by spontaneous evacuation were examined by direct microscopy and the Willis concentration method. A total of 150 individuals were infected (36.14%), 107 (25.78%) of whom with B. hominis. There was no difference between males and females (p > 0.05), but there was a significant difference between ages (chi(2) = 12.17; g.l. = 4), with infection more frequent between 18 and 27 years. In 71.02% of the cases, B. hominis was the only parasite. Giardia lamblia was the parasite most frequently associated with B. hominis (2.41%). In the majority (85%) of infected individuals, less than five microorganisms per microscopic field were observed. We conclude that B. hominis is a frequent intestinal parasite among food handlers in Caroní Municipality, Bolivar State, Venezuela.

  3. The prevalence of Blastocystis hominis and other protozoan parasites in soldiers returning from peacekeeping missions.

    PubMed

    Duda, Aleksandra; Kosik-Bogacka, Danuta; Lanocha-Arendarczyk, Natalia; Kołodziejczyk, Lidia; Lanocha, Aleksandra

    2015-04-01

    Blastocystis hominis is a common intestinal parasite found in humans living in poor sanitary conditions, living in tropical and subtropical climates, exposed to infected animals, or consuming contaminated food or water. The aim of this study was to determine the prevalence of B. hominis in Polish military personnel returning from peacekeeping missions in Iraq and Afghanistan. In total, 1,826 stool samples were examined. Gastrointestinal parasites were detected in 17% of the soldiers. The examined stool samples most frequently contained vacuolar forms of B. hominis (15.3%) and cysts of Entamoeba coli (1.0%) or Giardia lamblia (0.7%). In 97.1% of stool samples from infected soldiers, we observed less than five developmental forms of B. hominis in the field of view (40×). The parasite infections in soldiers were diagnosed in the autumn and the spring. There was no statistical correlation between age and B. hominis infection. Our results show that peacekeeping missions in countries with tropical or subtropical climates could be associated with risk for parasitic diseases, including blastocystosis. PMID:25732683

  4. The Prevalence of Blastocystis hominis and Other Protozoan Parasites in Soldiers Returning from Peacekeeping Missions

    PubMed Central

    Duda, Aleksandra; Kosik-Bogacka, Danuta; Lanocha-Arendarczyk, Natalia; Kołodziejczyk, Lidia; Lanocha, Aleksandra

    2015-01-01

    Blastocystis hominis is a common intestinal parasite found in humans living in poor sanitary conditions, living in tropical and subtropical climates, exposed to infected animals, or consuming contaminated food or water. The aim of this study was to determine the prevalence of B. hominis in Polish military personnel returning from peacekeeping missions in Iraq and Afghanistan. In total, 1,826 stool samples were examined. Gastrointestinal parasites were detected in 17% of the soldiers. The examined stool samples most frequently contained vacuolar forms of B. hominis (15.3%) and cysts of Entamoeba coli (1.0%) or Giardia lamblia (0.7%). In 97.1% of stool samples from infected soldiers, we observed less than five developmental forms of B. hominis in the field of view (40×). The parasite infections in soldiers were diagnosed in the autumn and the spring. There was no statistical correlation between age and B. hominis infection. Our results show that peacekeeping missions in countries with tropical or subtropical climates could be associated with risk for parasitic diseases, including blastocystosis. PMID:25732683

  5. Myiasis caused by Dermatobia hominis: countries with increased risk for travelers going to neotropic areas.

    PubMed

    Villalobos, Guiehdani; Vega-Memije, Maria Elisa; Maravilla, Pablo; Martinez-Hernandez, Fernando

    2016-10-01

    Here, we review the human botfly (Dermatobia hominis), which belongs to a group of Diptera generically known as "myiasis-causing flies," characterized by the ability of their larvae to develop in animal flesh. In addition to its medical and economic importance, there is an academic interest in this botfly because of its peculiar biology, particularly because a phoretic diptera is needed to complete the life cycle. The larvae penetrate the host's skin, causing furuncle-like lesions that are pruritic, painful, and resemble subcutaneous nodules, producing irreversible perforations in the skin. Although D. hominis is distributed from Mexico to Argentina, a review performed by our working group from 1999 to 2015 determined that the countries with the highest infection rates in travelers are Belize, Bolivia, and Brazil. Interestingly, infected men show a higher variation in the distribution of the lesions than in women. Many treatment schemes have been suggested, including the application of highly dense liquids to the lesion to cause anoxia in the D. hominis larvae. We showed, for the first time, a Bayesian inference between D. hominis and other myiasis-causing flies. The flies grouped into two main clusters according to their capacity to produce facultative and obligatory myiasis, and D. hominis was phylogenetically close to Cuterebra spp.

  6. [Severe haemolysis caused by Mycoplasma pneumoniae].

    PubMed

    Bohr, Anne Lisbeth; Aagaard, Thomas Granum; Birgens, Henrik; Søborg, Christian

    2015-10-01

    Mycoplasma pneumoniae is naturally resistant to betalactamase antibiotics but is sensitive to macrolides. Occasionally, infections with M. pneumoniae can lead to severe anaemia due to its ability to cause haemolysis when cold agglutination occurs. Increasing bacterial resistance to macrolid antibiotics is a growing concern worldwide. We present two cases where infection with M. pneumoniae caused severe haemolysis, one of which was macrolide-resistant.

  7. [Mycoplasma pneumoniae meningoencephalitis in a young adult].

    PubMed

    Del Castillo, Marcelo; D'Giano, Carlos; Goicoechea, María Teresa; Morello, Fernando; Salsamendi, Paz; Mora, Andrea

    2005-01-01

    Mycoplasma pneumoniae infections have extrapulmonary complications that involve the nervous system. The neurologic manifestations are diverse. Although the prognosis is usually favorable, the patients can undergo severe permanent sequelae. We present a young female adult with acute meningoencephalitis as a complication of a lower respiratory infection, which followed a benign course without neurologic sequelae.

  8. DNA repair in Mycoplasma gallisepticum

    PubMed Central

    2013-01-01

    Background DNA repair is essential for the maintenance of genome stability in all living beings. Genome size as well as the repertoire and abundance of DNA repair components may vary among prokaryotic species. The bacteria of the Mollicutes class feature a small genome size, absence of a cell wall, and a parasitic lifestyle. A small number of genes make Mollicutes a good model for a “minimal cell” concept. Results In this work we studied the DNA repair system of Mycoplasma gallisepticum on genomic, transcriptional, and proteomic levels. We detected 18 out of 22 members of the DNA repair system on a protein level. We found that abundance of the respective mRNAs is less than one per cell. We studied transcriptional response of DNA repair genes of M. gallisepticum at stress conditions including heat, osmotic, peroxide stresses, tetracycline and ciprofloxacin treatment, stationary phase and heat stress in stationary phase. Conclusions Based on comparative genomic study, we determined that the DNA repair system M. gallisepticum includes a sufficient set of proteins to provide a cell with functional nucleotide and base excision repair and mismatch repair. We identified SOS-response in M. gallisepticum on ciprofloxacin, which is a known SOS-inducer, tetracycline and heat stress in the absence of established regulators. Heat stress was found to be the strongest SOS-inducer. We found that upon transition to stationary phase of culture growth transcription of DNA repair genes decreases dramatically. Heat stress does not induce SOS-response in a stationary phase. PMID:24148612

  9. A Compendium for Mycoplasma pneumoniae.

    PubMed

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

    2016-01-01

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

  10. A Compendium for Mycoplasma pneumoniae

    PubMed Central

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

    2016-01-01

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

  11. Morphological diversity of Blastocystis hominis in sodium acetate-acetic acid-formalin-preserved stool samples stained with iron hematoxylin.

    PubMed

    MacPherson, D W; MacQueen, W M

    1994-01-01

    The objective of this investigation was to study the morphological characteristics of Blastocystis hominis in sodium acetate-acetic acid-Formalin-preserved stool samples. Routinely processed samples were examined for morphological detail, including size, shape, nuclear detail, and central body characteristics. Morphological findings revealing the importance of recognizing B. hominis in the diagnostic laboratory are described. PMID:7510311

  12. [Morphology of Blastocystis hominis in feces and evaluation of parasitological methods].

    PubMed

    Peréz de Suarez, E; Guzmán de Rondón, C

    1994-01-01

    A study of the morphology of Blastocystis hominis in stool in ninety-four cases humans is described as central body, ameba and granular form were found as previously described. In addition a "Globulose" form as a variation of granular form is first described. The central body form (96.8%) was the most abundant form. Three parasitological methods as direct microscopical examination sample with saline solution 0.85%, lugol, Sudan III, stained with Quensel, Iron hematoxylin and culture are evaluated to detection the Blastocystis hominis forms. Our results show that the direct microscopical examination (saline solution 0.85%, lugol, Sudan III, stained with Quensel) is the most sensitive and specific method than culture. The identification of Blastocystis hominis in stool difficult due to the diversity of shapes and size, which generate confusion with other intestinal protozoa and host cells. PMID:7557277

  13. Eradication of Blastocystis hominis prevents the development of symptomatic Hashimoto's thyroiditis: a case report.

    PubMed

    Rajič, Borko; Arapović, Jurica; Raguž, Kazimir; Bošković, Mladen; Babić, Senaida Marina; Maslać, Suzana

    2015-07-01

    In this case report we describe a 49 year-old man who presented with chronic urticaria, angioedema and soft stool consistency. During diagnostic examinations Hashimoto's thyroiditis was found even though the patient never had clear symptoms of this disease. Blastocystis hominis was isolated through a stool microbiologic examination, implicating that this parasite can cause the development of Hashimoto's thyroiditis and chronic urticaria. After two-weeks treatment with metronidazole the Blastocystis hominis was eradicated, then urticaria and angioedema disappeared. During the four years of follow-up, the patient presented without any symptoms, whereas thyroid hormones were normalized and anti-thyroid antibodies declined. For the first time in the literature we show that eradication of Blastocystis hominis can prevent the development of both symptomatic Hashimoto's thyroiditis and chronic urticaria. PMID:26230132

  14. Molecular identification of Sarcocystis hominis in native cattle of central Iran: a case report.

    PubMed

    Hajimohammadi, B; Eslami, G; Oryan, A; Zohourtabar, A; Pourmirzaei Tafti, H; Moghaddam Ahmadi, M

    2014-03-01

    Sarcocystis spp. are two-host protozoan parasites belonging to the phylum Apicomplexa. Among different known species of Sarcocystis in cattle, only Sarcocystis hominis is important from the public health viewpoint, because of its zoonotic characteristics. This study presents the first molecular identification of S. hominis in native cattle in central Iran. A sample of diaphragm muscle from a 6-year-old native cow slaughtered at Yazd Slaughterhouse, Yazd, central Iran, was collected in May 2013. DNA extraction was performed, using the salting-out method. DNA purification and precipitation were performed consecutively. The amplicon and digestion results were analyzed using agarose gel electrophoresis. A PCR product with 926 bp in length was obtained after amplification, and 376 bp and 550 bp in length after digestion that identified S. hominis. To the best of our knowledge, this study is the first of its kind to be reported from Iran. PMID:24862059

  15. Detection of Pentatrichomonas hominis in dogs using real-time PCR.

    PubMed

    Michalczyk, M; Sokół, R; Socha, P

    2015-01-01

    Trichomonadidae family is a protozoan occurring in different animal species. It inhabits the gastrointestinal and urinary tracts. P. hominis is rarely found in faecal samples of dogs, and its identification and differentiation from other trichomonads by light microscopy are difficult. Methods of molecular biology are the most effective in this case, because they confirm the presence of the specific species in animal organisms, irrespective of the protozoan form. The aim of this study was to find P. hominis in selected dog kennels in North-Eastern Poland. Forty-one faecal samples of dogs from 7 dog kennels were examined. The occurrence of P. hominis in 5 faecal samples of dogs with no symptoms of diarrhoea was the first one to be confirmed in Poland.

  16. 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

  17. Isolation of mycoplasmas from a buzzard, falcons and vultures.

    PubMed

    Poveda, J B; Giebel, J; Kirchhoff, H; Fernandez, A

    1990-10-01

    Thirteen mycoplasmas were isolated from a peregrine falcon (Falco peregrinus), two saker falcons (Falco cherrug), a buzzard (Buteo buteo), a black vulture (Aegypius monachus), and two griffon vultures (Gypsfuhus). Six of them could be identified: Mycoplasma gallinarum (three isolates), M. columborale (two isolates) and M. anatis (one isolate). The remaining seven isolates did not react with antisera against the known avian mycoplasma species in the indirect immunofluorescence and growth inhibition tests. They may represent new species.

  18. A novel medium devoid of ruminant peptone for high yield growth of Mycoplasma ovipneumoniae.

    PubMed

    Patel, Hiren; Mackintosh, David; Ayling, Roger D; Nicholas, Robin A J; Fielder, Mark D

    2008-03-18

    Mycoplasma ovipneumoniae is considered an emerging veterinary pathogen causing pneumonia in sheep and goats worldwide. Currently it has not been possible to define a growth medium that yields the maximum growth of M. ovipneumoniae within a short incubation period. Growth yields of M. ovipneumoniae in Eaton's medium are variable and not as consistently high as those seen with other Mycoplasma spp. This study investigated the ability of different M. ovipneumoniae field strains to grow in various media formulations, where PPLO broth was replaced by a vegetable protein source, and comparisons were made in terms of strain viability in Eaton's medium. Studies were also conducted to determine the optimal carbohydrate source for use in the M. ovipneumoniae medium. Generally, it was found that different strains showed good growth in all media tested, with growth yields at 24h in TSB-1 medium higher than those observed with Eaton's medium. Growth yields reached 10(8) to 10(9)cfu ml(-1) within 24h for particular field strains, with all strains achieving this growth level within 48-72h.

  19. [Pyomyositis, sacroiliitis and spondylodiscitis caused by Staphylococcus hominis in a immunocompetent woman].

    PubMed

    Gómez Rodríguez, N; Durán Muñoz, O

    2006-12-01

    In absence of risk factors, osteoarticular infections by coagulase-negative staphylococci are very infrequent. We described the case of a immunocompetent 73-year-old-woman that suffered pyomyositis, left sacroiliitis and spondylodiscitis involving the first and second thoracic vertebrae by Staphylococcus hominis. This multifocal infection occurred five-weeks after intramuscular administration of NSAI for treatment of low back pain associated with a herniated disc L4-L5. This is the first know case of a multifocal muscle skeletal infection by Staphylococcus hominis in a patient immunocompetent.

  20. [Dermatobia hominis furuncular myiasis in a man returning from Latin America: first imported case in Tunisia].

    PubMed

    Kaouech, E; Kallel, K; Belhadj, S; Chaker, E

    2010-04-01

    Human cutaneous myiasis is a common dermatosis in tropical zones. The purpose of this report is to describe the first imported case of furuncular myiasis caused by Dermatobia hominis (human botfly) in Tunisia. The patient was a man returning from Bolivia. Furuncular myiasis was suspected based on epidemiological data and clinical examination showing pruriginous elevated lesions. Diagnosis was confirmed by identification of Dermatobia hominis larvae. Treatment was based mainly on manual removal of larvae. Since furuncular myiasis is unknown in Tunisia, it is important to remember this parasitic disease in differential diagnosis in patients presenting boil-like inflammatory papules following travel to Latin America.

  1. Development and host compatibility of plasmids for two important ruminant pathogens, Mycoplasma bovis and Mycoplasma agalactiae.

    PubMed

    Sharma, Shukriti; Citti, Chistine; Sagné, Eveline; Marenda, Marc S; Markham, Philip F; Browning, Glenn F

    2015-01-01

    Mycoplasma bovis is a cause of pneumonia, mastitis, arthritis and otitis media in cattle throughout the world. However, despite its clinical significance, there is a paucity of tools to genetically manipulate it, impeding our capacity to further explore the molecular basis of its virulence. To address this limitation, we developed a series of homologous and heterologous replicable plasmids from M. bovis and M. agalactiae. The shortest replicable oriC plasmid based on the region downstream of dnaA in M. bovis was 247 bp and contained two DnaA boxes, while oriC plasmids based on the region downstream of dnaA in M. agalactiae strains 5632 and PG2 were 219 bp and 217 bp in length, respectively, and contained only a single DnaA box. The efficiency of transformation in M. bovis and M. agalactiae was inversely correlated with the size of the oriC region in the construct, and, in general, homologous oriC plasmids had a higher transformation efficiency than heterologous oriC plasmids. The larger pWholeoriC45 and pMM21-7 plasmids integrated into the genomic oriC region of M. bovis, while the smaller oriC plasmids remained extrachromosomal for up to 20 serial passages in selective media. Although specific gene disruptions were not be achieved in M. bovis in this study, the oriC plasmids developed here could still be useful as tools in complementation studies and for expression of exogenous genes in both M. bovis and M. agalactiae.

  2. Field-Applicable Recombinase Polymerase Amplification Assay for Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae.

    PubMed

    Liljander, Anne; Yu, Mingyan; O'Brien, Elizabeth; Heller, Martin; Nepper, Julia F; Weibel, Douglas B; Gluecks, Ilona; Younan, Mario; Frey, Joachim; Falquet, Laurent; Jores, Joerg

    2015-09-01

    Contagious caprine pleuropneumonia (CCPP) is a highly contagious disease caused by Mycoplasma capricolum subsp. capripneumoniae that affects goats in Africa and Asia. Current available methods for the diagnosis of Mycoplasma infection, including cultivation, serological assays, and PCR, are time-consuming and require fully equipped stationary laboratories, which make them incompatible with testing in the resource-poor settings that are most relevant to this disease. We report a rapid, specific, and sensitive assay employing isothermal DNA amplification using recombinase polymerase amplification (RPA) for the detection of M. capricolum subsp. capripneumoniae. We developed the assay using a specific target sequence in M. capricolum subsp. capripneumoniae, as found in the genome sequence of the field strain ILRI181 and the type strain F38 and that was further evidenced in 10 field strains from different geographical regions. Detection limits corresponding to 5 × 10(3) and 5 × 10(4) cells/ml were obtained using genomic DNA and bacterial culture from M. capricolum subsp. capripneumoniae strain ILRI181, while no amplification was obtained from 71 related Mycoplasma isolates or from the Acholeplasma or the Pasteurella isolates, demonstrating a high degree of specificity. The assay produces a fluorescent signal within 15 to 20 min and worked well using pleural fluid obtained directly from CCPP-positive animals without prior DNA extraction. We demonstrate that the diagnosis of CCPP can be achieved, with a short sample preparation time and a simple read-out device that can be powered by a car battery, in <45 min in a simulated field setting.

  3. Field-Applicable Recombinase Polymerase Amplification Assay for Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae.

    PubMed

    Liljander, Anne; Yu, Mingyan; O'Brien, Elizabeth; Heller, Martin; Nepper, Julia F; Weibel, Douglas B; Gluecks, Ilona; Younan, Mario; Frey, Joachim; Falquet, Laurent; Jores, Joerg

    2015-09-01

    Contagious caprine pleuropneumonia (CCPP) is a highly contagious disease caused by Mycoplasma capricolum subsp. capripneumoniae that affects goats in Africa and Asia. Current available methods for the diagnosis of Mycoplasma infection, including cultivation, serological assays, and PCR, are time-consuming and require fully equipped stationary laboratories, which make them incompatible with testing in the resource-poor settings that are most relevant to this disease. We report a rapid, specific, and sensitive assay employing isothermal DNA amplification using recombinase polymerase amplification (RPA) for the detection of M. capricolum subsp. capripneumoniae. We developed the assay using a specific target sequence in M. capricolum subsp. capripneumoniae, as found in the genome sequence of the field strain ILRI181 and the type strain F38 and that was further evidenced in 10 field strains from different geographical regions. Detection limits corresponding to 5 × 10(3) and 5 × 10(4) cells/ml were obtained using genomic DNA and bacterial culture from M. capricolum subsp. capripneumoniae strain ILRI181, while no amplification was obtained from 71 related Mycoplasma isolates or from the Acholeplasma or the Pasteurella isolates, demonstrating a high degree of specificity. The assay produces a fluorescent signal within 15 to 20 min and worked well using pleural fluid obtained directly from CCPP-positive animals without prior DNA extraction. We demonstrate that the diagnosis of CCPP can be achieved, with a short sample preparation time and a simple read-out device that can be powered by a car battery, in <45 min in a simulated field setting. PMID:26085615

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    PubMed

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

    2016-07-01

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

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

    PubMed

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

    2016-07-01

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

  7. Genetic Variation in Sialidase and Linkage to N-acetylneuraminate Catabolism in Mycoplasma synoviae

    PubMed Central

    May, Meghan; Brown, Daniel R.

    2008-01-01

    We explored the genetic basis for intraspecific variation in mycoplasmal sialidase activity that correlates with virulence, and its potentially advantageous linkage to nutrient catabolism. Polymorphism in N-acetylneuraminate scavenging and degradation genes (sialidase, N-acetylneuraminate lyase, N-acetylmannosamine kinase, N-acetylmannosamine-6-phosphate epimerase, N-acetylglucosamine-6-phosphate deacetylase, and glucosamine-6-phosphate deaminase) was evident among eight strains of the avian pathogen Mycoplasma synoviae. Most differences were single nucleotide polymorphisms, ranging from 0.34 ± 0.04 substitutions per 100 bp for N-acetylmannosamine kinase to 0.65 ± 0.03 for the single-copy sialidase gene nanI. Missense mutations were twice as common as silent mutations in nanI; 26% resulted in amino acids dissimilar to consensus; and there was a 12-base deletion near the nanI promoter in strain WVU1853T, supporting a complex genetic basis for differences in sialidase activity. Two strains had identical frameshifts in the N-acetylneuraminate lyase gene nanA, resulting in nonsense mutations, and both had downstream deletions in nanA. Such genetic lesions uncouple extracellular liberation of sialic acid from generation of fructose-6-phosphate and pyruvate via intracellular N-acetylneuraminate degradation. Retention of nanI by such strains, but not others in the M. synoviae phylogenetic cluster, is evidence that sialidase has an important non-nutritional role in the ecology of M. synoviae and certain other mycoplasmas. PMID:18490131

  8. Isolation of Ureaplasma diversum and mycoplasmas from genital tracts of beef and dairy cattle in Saskatchewan

    PubMed Central

    Mulira, Gershon L.; Saunders, J. Robert; Barth, Albert D.

    1992-01-01

    We report herein a survey in which cultures of bovine reproductive tracts for Ureaplasma diversum and mycoplasmas were carried out in order to better understand the role of these organisms in granular vulvitis (GV). Samples cultured were vulvar swabs from clinically normal cows or ones with GV, preputial swabs or raw semen from bulls, and abomasal contents of aborted fetuses. Ureaplasma diversum was isolated from 104 (43.3%) of 240 dairy cows, 32 (27.1%) of 118 beef cows, 43 (47.2%) of 91 beef heifers, 23 (67.6%) of 34 beef bulls, and three (60%) of five dairy bulls. Mycoplasmas were isolated from 18 (7.5%) dairy cows, two (1.6%) beef cows, three (8.8%) beef bulls, and one dairy bull. No isolation was made from 97 aborted fetuses. For 65 dairy cows and 30 beef heifers with vulvar lesions, the isolation rates for ureaplasmas of 62.5% and 69.7%, respectively, were significantly higher (X2) than those for normal animals (37.5% and 30.3%). On immunofluorescent serotyping of 137 of the 205 isolates, there were 66 in serogroup C (strain T44), 18 in serogroup B (strain D48), eight in serogroup A (strain A417 or strain 2312), 14 cross-reacting, and 31 that were not identified. It was concluded that U. diversum is commonly present in the lower reproductive tract of beef/dairy cattle in Saskatchewan and is associated with granular vulvitis. PMID:17423929

  9. An epornitic of Mycoplasma gallisepticum in turkeys.

    PubMed

    Mason, S J; Maiers, J D

    1984-01-01

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

  10. An epornitic of Mycoplasma gallisepticum in turkeys.

    PubMed

    Mason, S J; Maiers, J D

    1984-01-01

    A major epornitic of Mycoplasma gallisepticum occurred in the Monroe, North Carolina, area between January and June of 1983. The outbreak involved 304,000 turkeys of various ages, which were slaughtered in the eradication program at a cost of more than $550,000 to growers and poultry companies. An infected peafowl was the likely source of infection on the first farm. Traffic between farms by growers and company personnel was theorized to be the means of further spread. PMID:6487195

  11. Electrical Properties and Ultrastructure of Mycoplasma Membranes

    PubMed Central

    Carstensen, Edwin L.; Maniloff, Jack; Einolf, Charles W.

    1971-01-01

    Mycoplasma, in particular species laidlawii and gallisepticum, are found to have a very small, low frequency conductivity as would be predicted by the dielectric model for bacteria and their apparent lack of cell wall structure. Membrane capacitance values for the two organisms are both about 0.9 μF/cm2, although electron micrographs show that the membrane of M. gallisepticum is 20-40 A thicker than that of M. laidlawii. ImagesFigure 1Figure 2 PMID:5089915

  12. Immunohistochemical demonstration of Mycoplasma gallinarum and Mycoplasma gallinaceum in naturally infected hen oviducts.

    PubMed

    Martin de las Mulas, J; Fernández, A; Sierra, M A; Poveda, J B; Carranza, J

    1990-11-01

    Using indirect immunoperoxidase (IIP), peroxidase anti-peroxidase (PAP) and avidin biotin-peroxidase complex (ABC) immunohistochemical methods, Mycoplasma gallinarum and M gallinaceum antigens were demonstrated in ethanol-fixed paraffin-embedded oviduct sections from hens the eggs from which showed suboptimal hatchability. Specific immunoperoxidase staining was detected at the mucosa in the magnum portion of the oviduct. Optimal staining was achieved by applying the ABC method, though both IIP and PAP methods can also be used for diagnosis. Isolation and identification techniques gave similar results for the species of avian mycoplasmas involved.

  13. Epidemiological survey of Giardia spp. and Blastocystis hominis in an Argentinian rural community.

    PubMed

    Minvielle, Marta Cecilia; Pezzani, Betina Cecilia; Cordoba, María Alejandra; De Luca, María Marta; Apezteguia, María Carmen; Basualdo, Juan Angel

    2004-09-01

    The aim of this study was to relate personal data, socio-cultural and environmental characteristics, and the presence of symptoms/signs with the frequencies of Giardia spp. and Blastocystis hominis among a rural population in Buenos Aires Province, Argentina. Of the surveyed population (350), 3.7% were infected with only Giardia spp. or 22.9% with B. hominis, and 2.3% were infected with both protozoa. The frequency of infection according to sex; 6.1% of males were infected and 1.6% of females by Giardia spp., 26.7% and 19.5% by B. hominis, and 2.4% and 2.2% by both parasites, respectively. Giardia spp. was detected in only three adults (over 14 years), but B. hominis was more frequent in adults than in children. The prevalences of these protozoa in this community are lower than those reported by other Argentinean studies, which is probably associated with the low density of the studied population (5.95 inhab/km2). Statistical analysis revealed that a male sex, flooding of the home, the use of a latrine, and an abdominal pain were correlated with the presence of these parasites, which indicate the importance of these factors in rural communities. PMID:15381860

  14. Epidemiological survey of Giardia spp. and Blastocystis hominis in an Argentinian rural community

    PubMed Central

    Minvielle, Marta Cecilia; Pezzani, Betina Cecilia; Cordoba, María Alejandra; De Luca, María Marta; Apezteguia, María Carmen

    2004-01-01

    The aim of this study was to relate personal data, socio-cultural and environmental characteristics, and the presence of symptoms/signs with the frequencies of Giardia spp. and Blastocystis hominis among a rural population in Buenos Aires Province, Argentina. Of the surveyed population (350), 3.7% were infected with only Giardia spp. or 22.9% with B. hominis, and 2.3% were infected with both protozoa. The frequency of infection according to sex; 6.1% of males were infected and 1.6% of females by Giardia spp., 26.7% and 19.5% by B. hominis, and 2.4% and 2.2% by both parasites, respectively. Giardia spp. was detected in only three adults (over 14 years), but B. hominis was more frequent in adults than in children. The prevalences of these protozoa in this community are lower than those reported by other Argentinean studies, which is probably associated with the low density of the studied population (5.95 inhab/km2). Statistical analysis revealed that a male sex, flooding of the home, the use of a latrine, and an abdominal pain were correlated with the presence of these parasites, which indicate the importance of these factors in rural communities. PMID:15381860

  15. Kinetics of decolourisation and biotransformation of direct black 38 by C. hominis and P. stutzeri.

    PubMed

    Bafana, Amit; Devi, Sivanesan Saravana; Krishnamurthi, Kannan; Chakrabarti, Tapan

    2007-04-01

    In the present study, a consortium of Cardiobacterium hominis and Pseudomonas stutzeri was isolated from an effluent treatment plant of a textile industry, based on its ability to decolourise azo dyes including direct black 38 (DB38), a benzidine-based azo dye. The role of each culture in the decolourisation process was elucidated, and C. hominis was found to decolourise the dye. Although P. stutzeri could not decolourise the dye, it was found to synergistically enhance dye decolourisation activity of C. hominis by scavenging oxygen in the medium and creating an anaerobic condition (oxidation/reduction potential -440 mV), which is known to be necessary for azo dye decolourisation. Together, the cultures could decolourise 90.5% of 100 mg l(-1) DB38 within 24 h. Kinetics of DB38 decolourisation was also examined, and P. stutzeri was found to increase V (max) and K (m) of decolourisation activity of C. hominis by 3.6- and 3-fold, respectively. The study also revealed a pathway of DB38 degradation with the release of benzidine from DB38 and subsequent degradation of benzidine to 4-aminobiphenyl by the cultures.

  16. Mycoplasmas and cancer: focus on nucleoside metabolism

    PubMed Central

    Vande Voorde, Johan; Balzarini, Jan; Liekens, Sandra

    2014-01-01

    The standard of care for patients suffering cancer often includes treatment with nucleoside analogues (NAs). NAs are internalized by cell-specific nucleobase/nucleoside transporters and, after enzymatic activation (often one or more phosphorylation steps), interfere with cellular nucleo(s)(t)ide metabolism and DNA/RNA synthesis. Therefore, their efficacy is highly dependent on the expression and activity of nucleo(s)(t)ide-metabolizing enzymes, and alterations thereof (e.g. by down/upregulated expression or mutations) may change the susceptibility to NA-based therapy and/or confer drug resistance. Apart from host cell factors, several other variables including microbial presence may determine the metabolome (i.e. metabolite concentrations) of human tissues. Studying the diversity of microorganisms that are associated with the human body has already provided new insights in several diseases (e.g. diabetes and inflammatory bowel disease) and the metabolic exchange between tissues and their specific microbiota was found to affect the bioavailability and toxicity of certain anticancer drugs, including NAs. Several studies report a preferential colonization of tumor tissues with some mycoplasma species (mostly Mycoplasma hyorhinis). These prokaryotes are also a common source of cell culture contamination and alter the cytostatic activity of some NAs in vitro due to the expression of nucleoside-catabolizing enzymes. Mycoplasma infection may therefore bias experimental work with NAs, and their presence in the tumor microenvironment could be of significance when optimizing nucleoside-based cancer treatment. PMID:26417262

  17. Emergence of Cryptosporidium hominis Monkey Genotype II and Novel Subtype Family Ik in the Squirrel Monkey (Saimiri sciureus) in China

    PubMed Central

    Zhou, Ziyao; Zhong, Zhijun; Shen, Liuhong; Cao, Suizhong; Yu, Xingming; Hu, Yanchuan; Chen, Weigang; Peng, Gangneng

    2015-01-01

    A single Cryptosporidium isolate from a squirrel monkey with no clinical symptoms was obtained from a zoo in Ya’an city, China, and was genotyped by PCR amplification and DNA sequencing of the small-subunit ribosomal RNA (SSU rRNA), 70-kDa heat shock protein (HSP70), Cryptosporidium oocyst wall protein, and actin genes. This multilocus genetic characterization determined that the isolate was Cryptosporidium hominis, but carried 2, 10, and 6 nucleotide differences in the SSU rRNA, HSP70, and actin loci, respectively, which is comparable to the variations at these loci between C. hominis and the previously reported monkey genotype (2, 3, and 3 nucleotide differences). Phylogenetic studies, based on neighbor-joining and maximum likelihood methods, showed that the isolate identified in the current study had a distinctly discordant taxonomic status, distinct from known C. hominis and also from the monkey genotype, with respect to the three loci. Restriction fragment length polymorphisms of the SSU rRNA gene obtained from this study were similar to those of known C. hominis but clearly differentiated from the monkey genotype. Further subtyping was performed by sequence analysis of the gene encoding the 60-kDa glycoprotein (gp60). Maximum homology of only 88.3% to C. hominis subtype IdA10G4 was observed for the current isolate, and phylogenetic analysis demonstrated that this particular isolate belonged to a novel C. hominis subtype family, IkA7G4. This study is the first to report C. hominis infection in the squirrel monkey and, based on the observed genetic characteristics, confirms a new C. hominis genotype, monkey genotype II. Thus, these results provide novel insights into genotypic variation in C. hominis. PMID:26509708

  18. Emergence of Cryptosporidium hominis Monkey Genotype II and Novel Subtype Family Ik in the Squirrel Monkey (Saimiri sciureus) in China.

    PubMed

    Liu, Xuehan; Xie, Na; Li, Wei; Zhou, Ziyao; Zhong, Zhijun; Shen, Liuhong; Cao, Suizhong; Yu, Xingming; Hu, Yanchuan; Chen, Weigang; Peng, Gangneng

    2015-01-01

    A single Cryptosporidium isolate from a squirrel monkey with no clinical symptoms was obtained from a zoo in Ya'an city, China, and was genotyped by PCR amplification and DNA sequencing of the small-subunit ribosomal RNA (SSU rRNA), 70-kDa heat shock protein (HSP70), Cryptosporidium oocyst wall protein, and actin genes. This multilocus genetic characterization determined that the isolate was Cryptosporidium hominis, but carried 2, 10, and 6 nucleotide differences in the SSU rRNA, HSP70, and actin loci, respectively, which is comparable to the variations at these loci between C. hominis and the previously reported monkey genotype (2, 3, and 3 nucleotide differences). Phylogenetic studies, based on neighbor-joining and maximum likelihood methods, showed that the isolate identified in the current study had a distinctly discordant taxonomic status, distinct from known C. hominis and also from the monkey genotype, with respect to the three loci. Restriction fragment length polymorphisms of the SSU rRNA gene obtained from this study were similar to those of known C. hominis but clearly differentiated from the monkey genotype. Further subtyping was performed by sequence analysis of the gene encoding the 60-kDa glycoprotein (gp60). Maximum homology of only 88.3% to C. hominis subtype IdA10G4 was observed for the current isolate, and phylogenetic analysis demonstrated that this particular isolate belonged to a novel C. hominis subtype family, IkA7G4. This study is the first to report C. hominis infection in the squirrel monkey and, based on the observed genetic characteristics, confirms a new C. hominis genotype, monkey genotype II. Thus, these results provide novel insights into genotypic variation in C. hominis.

  19. In vitro antibiotic susceptibility of field isolates of Mycoplasma synoviae in Argentina.

    PubMed

    Cerdá, R O; Giacoboni, G I; Xavier, J A; Sansalone, P L; Landoni, M F

    2002-01-01

    Minimum inhibitory concentrations (MICs) were determined in vitro for 7 antibiotics (aivlosin, enrofloxacine, tylosin, tiamulin, kitasamycin, chlortetracycline, and oxytetracycline) against eight recent local Argentinean isolates and two standard strains of Mycoplasma synoviae. Aivlosin (3-acetyl-4"-isovaleryl tylosin tartrate), tylosin, and tiamulin showed the lowest MICs with MIC90s of 0.006, 0.012, and 0.05 microg/ml, respectively. Except one strain that showed resistant values to chlortetracycline (> or = 12.5 microg/ml), all the analyzed strains were susceptible in different degrees to all the antibiotics tested. In this study, the improved activity of the tylosin-derived drug, aivlosin, was confirmed because it showed, in most strains, MIC values half those for tylosin. PMID:11922338

  20. In vitro antibiotic susceptibility of field isolates of Mycoplasma synoviae in Argentina.

    PubMed

    Cerdá, R O; Giacoboni, G I; Xavier, J A; Sansalone, P L; Landoni, M F

    2002-01-01

    Minimum inhibitory concentrations (MICs) were determined in vitro for 7 antibiotics (aivlosin, enrofloxacine, tylosin, tiamulin, kitasamycin, chlortetracycline, and oxytetracycline) against eight recent local Argentinean isolates and two standard strains of Mycoplasma synoviae. Aivlosin (3-acetyl-4"-isovaleryl tylosin tartrate), tylosin, and tiamulin showed the lowest MICs with MIC90s of 0.006, 0.012, and 0.05 microg/ml, respectively. Except one strain that showed resistant values to chlortetracycline (> or = 12.5 microg/ml), all the analyzed strains were susceptible in different degrees to all the antibiotics tested. In this study, the improved activity of the tylosin-derived drug, aivlosin, was confirmed because it showed, in most strains, MIC values half those for tylosin.

  1. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... GENERAL BIOLOGICAL PRODUCTS STANDARDS Mycoplasma § 610.30 Test for Mycoplasma. Except as provided... produced from in vitro living cell cultures, and prior to inactivation in the case of inactivated virus vaccines produced from such living cell cultures, each virus harvest pool and control fluid pool shall...

  2. 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.

  3. 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

  4. Comparative antibiotic eradication of mycoplasma infections from continuous cell lines.

    PubMed

    Uphoff, Cord C; Drexler, Hans G

    2002-02-01

    Accumulating data implicate mycoplasma contamination as the single biggest problem in the culture of continuous cell lines. Mycoplasma infection can affect virtually every parameter and functional activity of the eukaryotic cells. A successful alternative to discarding infected cultures is to attempt to eliminate the contaminants by treatment with specific and efficient antimycoplasma antibiotics. The addition of antibiotics to the culture medium during a limited period of time (1-3 wk) is a simple, inexpensive, and very practical approach for decontaminating continuous cell lines. Here, we examined the effectiveness of several antibiotic treatment protocols that we have employed routinely in our cell lines bank. On an aggregate, 673 cultures from 236 chronically mycoplasma-positive cell lines were exposed to one of the following five antibiotic regimens: mycoplasma removal agent (quinolone; a 1-wk treatment), enrofloxacin (quinolone; 1 wk), sparfloxacin (quinolone; 1 wk), ciprofloxacin (quinolone; 2 wk), and BM-Cyclin (alternating tiamulin and minocycline; 3 wk). The mycoplasma infection was permanently (as determined by three solid mycoplasma detection assays) eliminated by the various antibiotics in 66-85% of the cultures treated. Mycoplasma resistance was seen in 7-21%, and loss of the culture as a result of cytotoxically caused cell death occurred in 3-11% of the cultures treated. Overall, 223 of the 236 mycoplasma-positive cell lines could be cured in a first round of antibiotic treatment with at least one regimen. Taken together, 95% of the mycoplasma-infected cell lines were permanently cleansed of the contaminants by antibiotic treatment, which validates this approach as an efficient and technically simple mycoplasma eradication method.

  5. Comparative antibiotic eradication of mycoplasma infections from continuous cell lines.

    PubMed

    Uphoff, Cord C; Drexler, Hans G

    2002-02-01

    Accumulating data implicate mycoplasma contamination as the single biggest problem in the culture of continuous cell lines. Mycoplasma infection can affect virtually every parameter and functional activity of the eukaryotic cells. A successful alternative to discarding infected cultures is to attempt to eliminate the contaminants by treatment with specific and efficient antimycoplasma antibiotics. The addition of antibiotics to the culture medium during a limited period of time (1-3 wk) is a simple, inexpensive, and very practical approach for decontaminating continuous cell lines. Here, we examined the effectiveness of several antibiotic treatment protocols that we have employed routinely in our cell lines bank. On an aggregate, 673 cultures from 236 chronically mycoplasma-positive cell lines were exposed to one of the following five antibiotic regimens: mycoplasma removal agent (quinolone; a 1-wk treatment), enrofloxacin (quinolone; 1 wk), sparfloxacin (quinolone; 1 wk), ciprofloxacin (quinolone; 2 wk), and BM-Cyclin (alternating tiamulin and minocycline; 3 wk). The mycoplasma infection was permanently (as determined by three solid mycoplasma detection assays) eliminated by the various antibiotics in 66-85% of the cultures treated. Mycoplasma resistance was seen in 7-21%, and loss of the culture as a result of cytotoxically caused cell death occurred in 3-11% of the cultures treated. Overall, 223 of the 236 mycoplasma-positive cell lines could be cured in a first round of antibiotic treatment with at least one regimen. Taken together, 95% of the mycoplasma-infected cell lines were permanently cleansed of the contaminants by antibiotic treatment, which validates this approach as an efficient and technically simple mycoplasma eradication method. PMID:11929000

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

    PubMed Central

    2014-01-01

    Background Hemotropic mycoplasmas are epicellular erythrocytic bacteria that can cause infectious anemia in some mammalian species. Worldwide, hemotropic mycoplasmas are emerging or re-emerging zoonotic pathogens potentially causing serious and significant health problems in wildlife. The objective of this study was to determine the molecular prevalence of hemotropic Mycoplasma species in little brown bats (Myotis lucifugus) with and without Pseudogymnoascus (Geomyces) destrucans, the causative agent of white nose syndrome (WNS) that causes significant mortality events in bats. Methods In order to establish the prevalence of hemotropic Mycoplasma species in a population of 68 little brown bats (Myotis lucifugus) with (n = 53) and without (n = 15) white-nose syndrome (WNS), PCR was performed targeting the 16S rRNA gene. Results The overall prevalence of hemotropic Mycoplasmas in bats was 47%, with similar (p = 0.5725) prevalence between bats with WNS (49%) and without WNS (40%). 16S rDNA sequence analysis (~1,200 bp) supports the presence of a novel hemotropic Mycoplasma species with 91.75% sequence homology with Mycoplasma haemomuris. No differences were found in gene sequences generated from WNS and non-WNS animals. Conclusions Gene sequences generated from WNS and non-WNS animals suggest that little brown bats could serve as a natural reservoir for this potentially novel Mycoplasma species. Currently, there is minimal information about the prevalence, host-specificity, or the route of transmission of hemotropic Mycoplasma spp. among bats. Finally, the potential role of hemotropic Mycoplasma spp. as co-factors in the development of disease manifestations in bats, including WNS in Myotis lucifugus, remains to be elucidated. PMID:24655520

  7. The humoral immune response of lambs experimentally infected with Mycoplasma ovipneumoniae.

    PubMed

    Thirkell, D; Spooner, R K; Jones, G E; Russell, W C

    1990-08-01

    Using sera from lambs experimentally infected with Mycoplasma ovipneumoniae and Pasteurella haemolytica, the development of a good humoral immune response to M. ovipneumoniae was detected by ELISA. The antibody titres peaked 41 days post-infection and good antibody titres were maintained over the 16-week experimental period. Immunoblotting revealed that antibodies to specific antigens appeared in the sera in a sequential manner, some being seen shortly after infection and others developing only after a substantial time lag. Antibodies were raised against almost all the major antigens detected in one laboratory strain (956/2) and against all antigens previously shown to be conserved in 22 Scottish field isolates of M. ovipneumoniae.

  8. Genetic diversity of Mycoplasma arginini isolates based on multilocus sequence typing.

    PubMed

    Olaogun, Olusola M; Kanci, Anna; Barber, Stuart R; Tivendale, Kelly A; Markham, Philip F; Marenda, Marc S; Browning, Glenn F

    2015-10-22

    The contribution of Mycoplasma arginini to mycoplasmosis in small ruminants remains unclear because it is recovered from both healthy and diseased animals. In order to gain a better understanding of any relationships between isolates from different sites and different geographical locations, we developed a method for genotyping M. arginini using multilocus sequence typing (MLST). A MLST scheme based on five housekeeping genes was used to characterize M. arginini isolates from flocks of sheep and goats. A high level of genetic variability was detected between strains and within herds. PMID:26264760

  9. Molecular characterisation of the Mycoplasma cynos haemagglutinin HapA.

    PubMed

    Kastelic, Saša; Cizelj, Ivanka; Narat, Mojca; Tozon, Nataša; Chalker, Victoria J; Lysnyansky, Inna; Spergser, Joachim; Benčina, Dušan

    2015-01-30

    Mycoplasma (M.) cynos is a proven pathogen of dogs causing respiratory infections including pneumonia. We examined 19 M. cynos strains isolated from different organs of dogs in Austria, Denmark and Israel. All strains agglutinated mammalian and chicken erythrocytes. Using erythrocytes of chickens or dogs as specific ligands we isolated an approximately 65 kDa protein from cell-free supernatants of 3 M. cynos strains, which showed an apparent capacity for haemagglutination. The N-terminal sequence of a 25 kDa fragment of this protein was identified as NNEMTPKVTVEAKSMELLLSVEK. The identical amino acid sequence is encoded by the gene MCYN_0308 in the genome of M. cynos C142. This gene belongs to a family of some 20 genes which encode putative lipoproteins with proline-rich regions (PRR) in the first third of their molecules. We termed the 65 kDa haemagglutinin HapA and sequenced hapA gene homologues of 16 M. cynos strains. Analyses of hapA gene homologues revealed similar but not identical sequences, some having insertions and/or deletions in the PRR. We produced a recombinant HapA protein (rHapA) and also mouse monoclonal antibodies (mAbs) recognizing HapA. However, enzyme immunoassays using native M. cynos colonies and mAbs 5G2 or 3B7 showed variable expression of HapA in all M. cynos strains. This was further confirmed by Western blot analyses which showed different HapA quantities and also size-variation of HapA among strains. Analyses of cDNA of the expressed hapA genes showed that besides the hapA gene cultures of M. cynos (strains 105, 2002, 2297) can also express other forms of hap genes. In addition, in cloned cultures of strain 2297 altered HapA epitopes for mAbs 5G2 and 3B7 with distinct hapA gene mutations that resulted in altered HapA amino acid sequence were found. Most of the dogs examined had serum antibodies to rHapA. In conclusion, we characterized the M. cynos haemagglutinin HapA protein and encoding gene hapA, a factor involved in cytadherence to

  10. Mycoplasma Contamination Revisited: Mesenchymal Stromal Cells Harboring Mycoplasma hyorhinis Potently Inhibit Lymphocyte Proliferation In Vitro

    PubMed Central

    Zinöcker, Severin; Wang, Meng-Yu; Gaustad, Peter; Kvalheim, Gunnar; Rolstad, Bent; Vaage, John T.

    2011-01-01

    Background Mesenchymal stromal cells (MSC) have important immunomodulatory effects that can be exploited in the clinical setting, e.g. in patients suffering from graft-versus-host disease after allogeneic stem cell transplantation. In an experimental animal model, cultures of rat T lymphocytes were stimulated in vitro either with the mitogen Concanavalin A or with irradiated allogeneic cells in mixed lymphocyte reactions, the latter to simulate allo-immunogenic activation of transplanted T cells in vivo. This study investigated the inhibitory effects of rat bone marrow-derived MSC subsequently found to be infected with a common mycoplasma species (Mycoplasma hyorhinis) on T cell activation in vitro and experimental graft-versus-host disease in vivo. Principal Findings We found that M. hyorhinis infection increased the anti-proliferative effect of MSC dramatically, as measured by both radiometric and fluorimetric methods. Inhibition could not be explained solely by the well-known ability of mycoplasmas to degrade tritiated thymidine, but likely was the result of rapid dissemination of M. hyorhinis in the lymphocyte culture. Conclusions This study demonstrates the potent inhibitory effect exerted by M. hyorhinis in standard lymphocyte proliferation assays in vitro. MSC are efficient vectors of mycoplasma infection, emphasizing the importance of monitoring cell cultures for contamination. PMID:21264307

  11. Antigenic and Genetic Characterization of Lipoprotein LppQ from Mycoplasma mycoides subsp. mycoides SC

    PubMed Central

    Abdo, El-Mostafa; Nicolet, Jacques; Frey, Joachim

    2000-01-01

    Lipoprotein LppQ, a predominant 48-kDa antigen, and its corresponding gene, lppQ, were characterized in Mycoplasma mycoides subsp. mycoides SC, the etiological agent of contagious bovine pleuropneumonia. The lppQ gene is specific to M. mycoides subsp. mycoides SC and was found in the type strain and in field strains isolated in Europe, Africa, and Australia, as well as in vaccinal strains. LppQ is encoded as a precursor with a consensus sequence for prokaryotic signal peptidase II and a lipid attachment site. The leader sequence shows significant prominent transmembrane helix structure with a predicted outside-to-inside helix formation capacity. The N-terminal domain of the mature LppQ was shown to be surface exposed. It induced a strong, specific, early, and persistent immune response in naturally and experimentally infected animals. The C-terminal domain of LppQ possesses an integral membrane structure built up of repeated units, rich in hydrophobic and aromatic amino acids, which have a pore formation potential. A recombinant peptide representing the N-terminal domain of LppQ was obtained by site-directed mutagenesis of nine Mycoplasma-specific TGA (Trp) codons into universal TGG (Trp) codons and expression in Escherichia coli hosts. It was used for serodetection of cattle infected with M. mycoides subsp. mycoides SC, in which it was detected postinfection for significantly longer than conventional serological test reactions. PMID:10882657

  12. Validation of a mycoplasma molecular diagnostic test and distribution of mycoplasma species in bovine milk among New York State dairy farms.

    PubMed

    Gioia, G; Werner, B; Nydam, D V; Moroni, P

    2016-06-01

    Mycoplasma mastitis is a contagious and costly disease of dairy cattle that significantly affects animal health and milk productivity. Mycoplasma bovis is the most prevalent and invasive agent of mycoplasma mastitis in dairy cattle, and early detection is critical. Other mycoplasma have been isolated from milk; however, the role and prevalence of these species as mastitis pathogens are poorly understood. Routine screening of milk for mycoplasma by bacteriological culture is an important component of a farm control strategy to minimize a herd mycoplasma outbreak, but phenotypic methods have limited ability to speciate mycoplasma, affecting how farms and practitioners can understand the role and effect of species other than M. bovis in herd health. Fastidious mycoplasma culture can be lengthy and inconclusive, resulting in delayed or false negative reports. We developed and validated a multitarget PCR assay that can in the same day confirm or reject a presumptive positive mycoplasma culture found upon bacteriological testing of clinical specimens, further discriminate between Acholeplasma and Mycoplasma, and identify M. bovis. Coupled with sequence analysis isolates can be further identified as bovine mycoplasma Mycoplasma arginini, Mycoplasma alkalescens, Mycoplasma canadense, Mycoplasma bovirhinis, Mycoplasma bovigenitalium, Mycoplasma californicum, Acholeplasma laidlawii, and Acholeplasma oculi. Assay validation included analysis of 845 mycoplasma representing these species and 30 additional bacterial species obtained from routine milk submissions to the Quality Milk Production Services from New York State farms and veterinary clinics between January 2012 and December 2015. Among 95 herds, we found 8 different Mycoplasma species and 3 different Acholeplasma species, with an overall prevalence of M. bovirhinis of 1%, A. oculi of 2%, M. arginini of 2%, M. californicum of 3%, M. canadense of 10%, M. bovigenitalium of 10%, A. laidlawii of 11%, M. alkalescens of 17

  13. Prevalence of mycoplasma antibodies in lesser prairie-chicken sera.

    PubMed

    Hagen, Christian A; Crupper, Scott S; Applegate, Roger D; Robel, Robert J

    2002-01-01

    Serologic testing by the serum plate agglutination (SPA) procedure was performed to detect the presence of cross-reacting antibodies to Mycoplasma meleagridis, Mycoplasma synoviae, and Mycoplasma gallisepticum in lesser prairie-chickens (Tympanuchus pallidicinctus) trapped over a 2-yr period in Finney and Kearny counties of southwestern Kansas. Sera examined from birds (n = 50) obtained in March-April 2000 tested positive for M meleagridis, M. synoviae, and M. gallisepticum at levels of 6%, 10%, and 10%, respectively, for the population examined. Mycoplasma meleagridis antibodies were detected in 3 samples (2.7%), M. synoviae antibodies in 2 samples (1.7%), and M. gallisepticum antibodies in 3 samples (2.7%) from birds (n = 112) collected in March-April 2001. Data obtained by SPA can result in false positives and should be verified by additional procedures such as the hemagglutination-inhibition test. Low amounts of sera prohibited this additional testing. Thus, the positive SPA results should be considered presumptive for the presence of Mycoplasma antibodies. Although Mycoplasma antibodies have been detected in wild turkeys (Meleagris gallopavo) from Kingman and Butler counties in Kansas, this report is the first of possible mycoplasmosis in Finney and Kearny counties, Kansas. All birds testing positive by this procedure should be considered as potential carriers of Mycoplasma and should not be used in relocation efforts.

  14. Cloning of the complete Mycoplasma pneumoniae genome.

    PubMed Central

    Wenzel, R; Herrmann, R

    1989-01-01

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

  15. [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

  16. 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.

  17. Morphology of Ureaplasma urealyticum (T-mycoplasma) organisms and colonies.

    PubMed Central

    Razin, S; Masover, G K; Palant, M; Hayflick, L

    1977-01-01

    The morphology of Ureaplasm urealyticum in broth cultures was studied by phase-contrast microscopy. Most organisms appeared singly or in pairs. Long filaments and long chains of cocci, common in classical mycoplasma cultures, were not observed. On solid medium, U. urealyticum produced "fried-egg" colonies which developed according to the scheme suggested by Razin and Oliver (J. Gen. Microbiol., 1961) for the morphogenesis of the classical mycoplasma colonies. The formation of the peripheral zone of the colonies followed that of the central zone only when growth conditions were adequate, Hence, the appearance of peripheral zones, and consequently the larger colony size, can be taken as an indicator of improved growth conditions. Incubation in an atmosphere of 100% CO2 resulted in significantly larger colonies than in an atmosphere of N2, O2, or air. CO2 acts as a buffer, keeping the pH at the optimal range for Ureaplasma growth (pH 6.0 to 6.5) in the presence of the ammonia produced from the urea hydrolyzed by the organisms. The addition to the medium of 0.01 M urea together with 0.01 M putrescine enabled better growth than with urea alone. Small amounts of phosphate improved growth in an atmosphere of CO2, apparently fulfilling a nutritional role. Under nitrogen, higher phosphate concentrations were required for good growth, apparently serving as a buffer as well as a nutrient. Sodium chloride and sucrose which had been added to increase the tonicity of the medium inhibited growth above 0.1 M. An increase in the agar concentration above 2% resulted in decreased colony size. Likewise, prolonged drying of the agar plates caused a marked decrease in colony size, mostly affecting the peripheral zone. The addition of both urea and putrescine to the growth medium and incubation in a humidified CO2 atmosphere are recommended for improved growth and formation of fried-egg colonies of U. ureaplyticum on agar. It must be emphasized that these experiments were carried out

  18. The first reported cases of human cryptosporidiosis caused by Cryptosporidium hominis in Slovak Republic.

    PubMed

    Ondriska, František; Vrabcová, Ivana; Brinďáková, Silvia; Kváč, Martin; Ditrich, Oleg; Boldiš, Vojtech; Bastlová, Marcela

    2013-01-01

    Cryptosporidiosis belongs to the important parasitic infections with zoonotic potential and the occurrence in European countries is rare. The first cases of cryptosporidiosis caused by Cryptosporidium hominis detected in the Slovak republic were described here. Collection of examined humans consisted of five family members. Faecal specimens were examined by formalin sedimentation, by the Sheather's sugar flotation and by immunochromatography and visualised by the Ziehl-Neelsen acid fast stain. A fragment of the Cryptosporidium small subunit ribosomal RNA gene was amplified by nested polymerase chain reaction and species was determined by restriction fragment length polymorphism analysis with the endonucleases SspI and VspI. C. hominis was found in faeces of two immunocompetent siblings (a 7-year-old boy and a 2-year-old girl). The symptoms occurred only in the boy as gastrointestinal disorders lasting 5 days, and manifested by abdominal pain, an elevated body temperature (37.2 °C), mild diarrhoea, accompanied by lassitude, depression and anorexia. Ultrasonic scan revealed enlarged spleen and mezenteric lymph nodes. Microscopic examination of the stool sample revealed numerous Cryptosporidium oocysts. The DNA typing identified C. hominis subtype IbA10G2. Cryptosporidium was also detected in the boy's sister without any complications and symptoms. Their father, mother and grandmother were parasitologically negative. The source of infection remained unknown. Human cases in present study reflect necessity of systematic attention on intestinal parasites diagnostic inclusive of cryptosporidia.

  19. Genetic recombination and Cryptosporidium hominis virulent subtype IbA10G2.

    PubMed

    Li, Na; Xiao, Lihua; Cama, Vitaliano A; Ortega, Ynes; Gilman, Robert H; Guo, Meijin; Feng, Yaoyu

    2013-10-01

    Little is known about the emergence and spread of virulent subtypes of Cryptosporidium hominis, the predominant species responsible for human cryptosporidiosis. We conducted sequence analyses of 32 genetic loci of 53 C. hominis specimens isolated from a longitudinally followed cohort of children living in a small community. We identified by linkage disequilibrium and recombination analyses only limited genetic recombination, which occurred exclusively within the 60-kDa glycoprotein gene subtype IbA10G2, a predominant subtype for outbreaks in industrialized nations and a virulent subtype in the study community. Intensive transmission of virulent subtype IbA10G2 in the study area might have resulted in genetic recombination with other subtypes. Moreover, we identified selection for IbA10G2 at a 129-kb region around the 60-kDa glycoprotein gene in chromosome 6. These findings improve our understanding of the origin and evolution of C. hominis subtypes and the spread of virulent subtypes.

  20. Antibody Response to Mycoplasma pneumoniae: Protection of Host and Influence on Outbreaks?

    PubMed Central

    Dumke, Roger; Jacobs, Enno

    2016-01-01

    In humans of all ages, the cell wall-less and genome-reduced species Mycoplasma pneumoniae can cause infections of the upper and lower respiratory tract. The well-documented occurrence of major peaks in the incidence of community-acquired pneumonia cases reported world-wide, the multifaceted clinical manifestations of infection and the increasing number of resistant strains provide reasons for ongoing interest in the pathogenesis of mycoplasmal disease. The results of recent studies have provided insights into the interaction of the limited virulence factors of the bacterium with its host. In addition, the availability of complete M. pneumoniae genomes from patient isolates and the development of proteomic methods for investigation of mycoplasmas have not only allowed characterization of sequence divergences between strains but have also shown the importance of proteins and protein parts for induction of the immune reaction after infection. This review focuses on selected aspects of the humoral host immune response as a factor that might influence the clinical course of infections, subsequent protection in cases of re-infections and changes of epidemiological pattern of infections. The characterization of antibodies directed to defined antigens and approaches to promote their induction in the respiratory mucosa are also preconditions for the development of a vaccine to protect risk populations from severe disease due to M. pneumoniae. PMID:26858711

  1. Inflammation-inducing Factors of Mycoplasma pneumoniae

    PubMed Central

    Shimizu, Takashi

    2016-01-01

    Mycoplasma pneumoniae, which causes mycoplasmal pneumonia in human, mainly causes pneumonia in children, although it occasionally causes disease in infants and geriatrics. Some pathogenic factors produced by M. pneumoniae, such as hydrogen peroxide and Community-Acquired Respiratory Distress Syndrome (CARDS) toxin have been well studied. However, these factors alone cannot explain this predilection. The low incidence rate of mycoplasmal pneumonia in infants and geriatrics implies that the strong inflammatory responses induced by M. pneumoniae coordinate with the pathogenic factors to induce pneumonia. However, M. pneumoniae lacks a cell wall and does not possess an inflammation-inducing endotoxin, such as lipopolysaccharide (LPS). In M. pneumoniae, lipoproteins were identified as an inflammation-inducing factor. Lipoproteins induce inflammatory responses through Toll-like receptors (TLR) 2. Because Mycoplasma species lack a cell wall and lipoproteins anchored in the membrane are exposed, lipoproteins and TLR2 have been thought to be important for the pathogenesis of M. pneumoniae. However, recent reports suggest that M. pneumoniae also induces inflammatory responses also in a TLR2-independent manner. TLR4 and autophagy are involved in this TLR2-independent inflammation. In addition, the CARDS toxin or M. pneumoniae cytadherence induces inflammatory responses through an intracellular receptor protein complex called the inflammasome. In this review, the inflammation-inducing factors of M. pneumoniae are summarized. PMID:27065977

  2. Mycoplasma biofilms ex vivo and in vivo.

    PubMed

    Simmons, Warren L; Dybvig, Kevin

    2009-06-01

    Biofilms are communities of microorganisms that are encased in polymeric matrixes and grow attached to biotic or abiotic surfaces. Despite their enhanced ability to resist antimicrobials and components of the immune system in vitro, few studies have addressed the interactions of biofilms with the host at the organ level. Although mycoplasmas have been shown to form biofilms on glass and plastic surfaces, it has not been determined whether they form biofilms on the tracheal epithelium. We developed a tracheal organ-mounting system that allowed the entire surface of the tracheal lumen to be scanned using fluorescence microscopy. We observed the biofilms formed by the murine respiratory pathogen Mycoplasma pulmonis on the epithelium of trachea in tracheal organ culture and in experimentally infected mice and found similar structure and biological characteristics as biofilms formed in vitro. This tracheal organ-mounting system can be used to study interactions between biofilms formed by respiratory pathogens and the host epithelium and to identify the factors that contribute to biofilm formation in vivo.

  3. Molecular Epidemiology of Mycoplasma conjunctivae in Caprinae: Transmission across Species in Natural Outbreaks

    PubMed Central

    Belloy, Luc; Janovsky, Martin; Vilei, Edy M.; Pilo, Paola; Giacometti, Marco; Frey, Joachim

    2003-01-01

    Mycoplasma conjunctivae is the etiological agent of infectious keratoconjunctivitis, a highly contagious ocular infection that affects both domestic and wild Caprinae species in the European Alps. In order to study the transmission and spread of M. conjunctivae across domestic and wild Caprinae populations, we developed a molecular method for subtyping and identifying strains of M. conjunctivae. This method is based on DNA sequence determination of a variable domain within the gene lppS, a gene that encodes an antigenic lipoprotein of M. conjunctivae. This domain of lppS shows variations among different strains but remains constant upon generations of individual strains on growth medium and thus allows identification of individual strains and estimation of their phylogenetic intercorrelations. The variable domain of lppS is amplified by PCR using primers that match conserved sequences of lppS flanking it. Sequence analysis of the amplified fragment enables fine subtyping of M. conjunctivae strains. The method is applicable both to isolated strains and to clinical samples directly without requiring the cultivation of the strain. Using this method, we show that M. conjunctivae was transmitted between domestic and wild animals that were grazing in proximate pastures. Certain animals also presented infections with two different strains simultaneously. PMID:12676664

  4. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... inoculated in evenly distributed amounts over the surface of no less than 10 plates of at least two agar... the agar shall be excised from the inoculated area and examined for the presence of Mycoplasma....

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

    SciTech Connect

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

    1985-06-01

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

  6. Development of a specific DNA probe and PCR for the detection of Mycoplasma bovis.

    PubMed

    Ghadersohi, A; Coelen, R J; Hirst, R G

    1997-05-01

    Mycoplasma bovis is responsible for several production diseases in cattle, including mastitis, arthritis, pneumonia, abortion and infertility. Current methodologies for detecting and identifying M. bovis are time consuming and difficult. Tests which rely on antigen or antibody detection have poor sensitivity and specificity. In this paper associated protocols for the development of a hybridization probe and PCR are described. A genomic library (SauIIIA digested) was prepared from M. bovis DNA (Colindale Reference Strain: NC10131:02) and cloned into pUC19. Colony hybridization, using a probe preparation made from purified M. bovis DNA, was used to identify colonies of interest. M. bovis DNA fragments were retrieved from recombinant plasmids by digestion with EcoRI and HindIII. This DNA was used to prepare randomly primed probes for dot blot hybridization analysis with immobilized DNA from M. bovis (two strains), M. dispar, M. agalactiae, M. bovigenitalium (two strains), M. ovipneumoniae, a Group 7 strain, M. arginini and bacteria belonging to different genera. Four probes were found to hybridize only with M. bovis and M. ovipneumoniae DNA, whereas one probe reacted with genomic DNA from only one of the two M. bovis strains. The level of sensitivity of the dot blot hybridization assay was 200 CFU (colony forming units)/mL. To enhance the sensitivity further, an M. bovis-specific PCR assay was developed. The primers were designed using sequences obtained from the probe DNA which discriminated M. bovis from all other Mycoplasma DNA tested. The minimum amount of target DNA that could be detected by the PCR assay was that isolated from 10-20 CFU/mL. The PCR assay was therefore 10 times more sensitive than dot blot hybridization.

  7. Mycoplasma species and related organisms isolated from ruminants in Britain between 1990 and 2000.

    PubMed

    Ayling, R D; Bashiruddin, S E; Nicholas, R A J

    2004-10-01

    Between 1990 and 2000, more than 1600 mycoplasmas and the related acholeplasmas were identified from ruminant animals by the Mycoplasma Group at the Veterinary Laboratories Agency--Weybridge. Mycoplasma bovis was the most commonly identified pathogen, mostly from pneumonic calves but occasionally from cattle with mastitis and arthritis. Mycoplasma canis was first isolated in Britain in 1995 from pneumonic calves and the number of isolates increased to 18 per cent of the total mycoplasmas isolated from cattle in 1999. The ELISA for antibodies to M. bovis detected 1971 positive samples (22 per cent) among 8959 serum samples, mainly from pneumonic cattle. Other mycoplasmas identified included Mycoplasma dispar from the lungs of cattle with respiratory disease, and Mycoplasma bovigenitalium from the reproductive tract of cows with vulvovaginitis and infertility. Mycoplasma bovirhinis and Acholeplasma species were found commonly but are thought to be more opportunistic than pathogenic. In sheep and goats, the majority of Mycoplasma species isolated were identified as Mycoplasma ovipneumoniae from pneumonic sheep, Mycoplasma conjunctivae from sheep with keratoconjunctivitis, and the ubiquitous Mycoplasma arginini.

  8. Molecular Epidemiology of Methicillin-Resistant Staphylococcus hominis (MRSHo): Low Clonality and Reservoirs of SCCmec Structural Elements

    PubMed Central

    Bouchami, Ons; Ben Hassen, Assia; de Lencastre, Herminia; Miragaia, Maria

    2011-01-01

    Background Methicillin resistant Staphylococcus hominis (MRSHo) are important human pathogens in immunocompromised patients. However, little is known regarding its population structure and staphylococcal chromosomal cassette mec (SCCmec) content. Methodology/Principal Findings To assess the population structure and the SCCmec content of S. hominis, 34 MRSHo and 11 methicillin-susceptible S. hominis (MSSHo) from neutropenic patients collected over a 3-year period were studied. The genetic backgrounds of S. hominis isolates were analyzed by pulsed-field gel electrophoresis (PFGE) and SCCmec types were determined by PCR. Cassette chromosome recombinases (ccr) were characterized by PCR and ccrB sequencing. The 34 S. hominis isolates were classified into as many as 28 types and 32 subtypes (SID = 99.82%); clonal dissemination was occasionally observed. The main SCCmec structures identified were SCCmec type VI (4B) (20%), SCCmec VIII (4A) (15%), and a new SCCmec composed of mec complex A in association with ccrAB1 (38%); 27% of the isolates harbored non-typeable SCCmec. Overall, a high prevalence of mec complex A (73.5%), ccrAB1 (50%) and ccrAB4 (44%) were found. Importantly, ccrB1 and ccrB4 from both MRSHo and MSSHo showed a high nucleotide sequence homology with those found in S. aureus SCCmec I, VI and VIII respectively (>95%). Conclusions/Significance The S. hominis population showed a limited clonality and a low genetic diversity in the allotypes of ccr and classes of mec complex. Moreover, our data suggest that S. hominis might have been a privileged source of mec complex A, ccrB1 and ccrB4, for the assembly of primordial SCCmec types. PMID:21760926

  9. Molecular characterization of Mycoplasma arthritidis variable surface protein MAA2.

    PubMed

    Washburn, L R; Weaver, K E; Weaver, E J; Donelan, W; Al-Sheboul, S

    1998-06-01

    Earlier studies implied a role for Mycoplasma arthritidis surface protein MAA2 in cytadherence and virulence and showed that it exhibited both size and phase variability. Here we report the further analysis of MAA2 and the cloning and sequencing of the maa2 gene from two M. arthritidis strains, 158p10p9 and H606, expressing two size variants of MAA2. Triton X-114 partitioning and metabolic labeling with [3H]palmitic acid suggested lipid modification of MAA2. Surface exposure of the C terminus was indicated by cleavage of monoclonal antibody-specific epitopes from intact cells by carboxypeptidase Y. The maa2 genes from both strains were highly conserved, consisting largely of six (for 158p10p9) or five (for H606) nearly identical, 264-bp tandem direct repeats. The deduced amino acid sequence predicted a largely hydrophilic, highly basic protein with a 29-amino-acid lipoprotein signal peptide. The maa2 gene was expressed in Escherichia coli from the lacZ promoter of vector pGEM-T. The recombinant product was approximately 3 kDa larger than the native protein, suggesting that the signal peptide was not processed in E. coli. The maa2 gene and upstream DNA sequences were cloned from M. arthritidis clonal variants differing in MAA2 expression state. Expression state correlated with the length of a poly(T) tract just upstream of a putative -10 box. Full-sized recombinant MAA2 was expressed in E. coli from genes derived from both ON and OFF expression variants, indicating that control of expression did not include alterations within the coding region.

  10. Assessment of polymorphic genetic markers for multi-locus typing of Cryptosporidium parvum and Cryptosporidium hominis.

    PubMed

    Robinson, Guy; Chalmers, Rachel M

    2012-10-01

    The use of high resolution molecular tools to study Cryptosporidium parvum and Cryptosporidium hominis intra-species variation is becoming common practice, but there is currently no consensus in the methods used. The most commonly applied tool is partial gp60 gene sequence analysis. However, multi-locus schemes are acknowledged to improve resolution over analysis of a single locus, which neglects potential re-assortment of genes during the sexual phase of the Cryptosporidium life-cycle. Multi-locus markers have been investigated in isolates from a variety of sampling frames, in varying combinations and using different assays and methods of analysis. To identify the most informative markers as candidates for the development of a standardised multi-locus fragment size-based typing (MLFT) scheme to integrate with epidemiological analyses, we examined the published literature. A total of 31 MLFT studies were found, employing 55 markers of which 45 were applied to both C. parvum and C. hominis. Of the studies, 11 had sufficient raw data, from three or more markers, and a sampling frame containing at least 50 samples, for meaningful in-depth analysis using assessment criteria based on the sampling frame, study size, number of markers investigated in each study, marker characteristics (>2 nucleotide repeats) and the combinations of markers generating all possible multi-locus genotypes. Markers investigated differed between C. hominis and C. parvum. When each scheme was analysed for the fewest markers required to identify 95% of all MLFTs, some redundancy was identified in all schemes; an average redundancy of 40% for C. hominis and 27% for C. parvum. Ranking markers, based on the most productive combinations, identified two different sets of potentially most informative candidate markers, one for each species. These will be subjected to technical evaluation including typability (percentage of samples generating a complete multi-locus type) and discriminatory power by

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

    PubMed

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

    2015-04-01

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

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

    PubMed

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

    2015-04-01

    Mycoplasma infections can cause significant morbidity and mortality in captive and wild chelonians. As part of a health assessment of endangered bog turtles (Glyptemys muhlenbergii) in the northeastern US, choanal and cloacal swabs from these and other sympatric species, including spotted turtles (Clemmys guttata), eastern box turtles (Terrapene carolina carolina), wood turtles (Glyptemys insculpta), and common snapping turtles (Chelydra serpentina) from 10 sampling sites in the states (US) of Delaware, New Jersey, and Pennsylvania, were tested by PCR for Mycoplasma. Of 108 turtles tested, 63 (58.3%) were PCR positive for Mycoplasma including 58 of 83 bog turtles (70%), three of three (100%) eastern box turtles, and two of 11 (18%) spotted turtles; all snapping turtles (n = 7) and wood turtles (n = 4) were negative. Sequence analysis of portions of the 16S-23S intergenic spacer region and the 16S ribosomal RNA gene revealed a single, unclassified species of Mycoplasma that has been previously reported in eastern box turtles, ornate box turtles (Terrapene ornata ornata), western pond turtles (Emys marmorata), and red-eared sliders (Trachemys scripta elegans). We document a high incidence of Mycoplasma, in the absence of clinical disease, in wild emydid turtles. These findings, along with wide distribution of the identified Mycoplasma sp. across a broad geographic region, suggest this bacterium is likely a commensal inhabitant of bog turtles, and possibly other species of emydid turtles, in the northeastern US. PMID:25574806

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2016-01-01

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

  16. Comparison of Mycoplasma ovipneumoniae isolates using bacterial restriction endonuclease DNA analysis and SDS-PAGE.

    PubMed

    Mew, A J; Ionas, G; Clarke, J K; Robinson, A J; Marshall, R B

    1985-12-01

    Sixteen isolates of Mycoplasma ovipneumoniae recovered from the nasal tract or lungs of sheep from different flocks in New Zealand were examined by bacterial restriction endonuclease DNA analysis (BRENDA) using EcoR1 and by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). All isolates gave BRENDA patterns which differed entirely from one another. Following 20 serial passages (corresponding to approximately 67 generations) of an isolate, no change was detected in the BRENDA pattern. When eight isolates were examined by SDS-PAGE most bands were common but, nevertheless, each isolate was unique in the sense that they differed from one another in one or more bands. The marked heterogeneity of patterns observed when strains of M. ovipneumoniae are compared by BRENDA, together with the stability of such patterns over many generations, will enable this approach to be used to study the epidemiology of individual strains of M. ovipneumoniae within a flock.

  17. Persistence of Mycoplasma hyopneumoniae in experimentally infected pigs after marbofloxacin treatment and detection of mutations in the parC gene.

    PubMed

    Le Carrou, J; Laurentie, M; Kobisch, M; Gautier-Bouchardon, A V

    2006-06-01

    The ability of Mycoplasma hyopneumoniae to persist despite fluoroquinolone treatments was investigated with pigs. Groups of specific-pathogen-free pigs were experimentally infected with M. hyopneumoniae strain 116 and treated with marbofloxacin at the therapeutic dose (TD) or half of the therapeutic dose (TD/2) for 3 days. Results showed that, despite tissue penetration of marbofloxacin, particularly in the trachea and the tracheal secretions, the treatments did not have any influence on M. hyopneumoniae recovery from tracheal swabs. Mycoplasmas were also isolated from inner organs and tissues such as liver, spleen, kidneys, and bronchial lymph nodes. Recontamination of pigs via environment could not explain mycoplasma persistence after medication, as decontamination of pigs and allocation to a new disinfected environment did not have any significant effect on the phenomenon. A significant decrease in the susceptibility level to marbofloxacin of 12 mycoplasma clones reisolated after the treatments (TD/2 and TD) was observed. Two point mutations were found in the ParC quinolone resistance-determining region (QRDR) of DNA topoisomerase IV (Ser80-->Phe and Asp84-->Asn), and one point mutation was observed just behind the QRDR of ParC (Ala116-->Glu). This is the first time that mutations in a gene coding for topoisomerase IV have been described for M. hyopneumoniae after in vivo marbofloxacin treatments in experimentally infected pigs. However, development of resistance is not sufficient to explain M. hyopneumoniae persistence in vivo since (i) marbofloxacin concentrations were above the marbofloxacin MIC of the wild-type strain and (ii) mycoplasmas reisolated after a single injection of marbofloxacin did not display an increased marbofloxacin MIC.

  18. The effect of Mycoplasma and mycoplasma removal agent on the hydrolase activity in fibroblasts of patients with lysosomal diseases.

    PubMed

    Souza, F T S; Sostruznik, L S; Scolari, R C; Castro, K J M; Andrade, C V; Giugliani, R; Coelho, J C

    2010-01-01

    This study was designed to evaluate the effect of mycoplasma contamination on acid hydrolase activity and the action of the mycoplasma removal agent (MRA), in cultures of human fibroblasts from individuals with lysosomal diseases. For this purpose, we measured the activity of the b-galactosidase, arylsulphatase B (ASB), hexosaminidase A and a-glucosidase enzymes. The activity of the above mentioned enzymes in fibroblasts contaminated by mycoplasma was measured before and after the addition of the MRA. The results were then compared to the enzymatic activity in contamination-free cultures. Only the ASB enzyme showed significant alteration in activity both in the presence of mycoplasma and MRA. The remaining enzymes did not suffer significant interference by the presence of the two agents. Of the four enzymes tested, three did not suffer significant alterations by the presence of the mycoplasma nor from the MRA. However, the activity measured in the ASB enzyme increased significantly in the presence of mycoplasma and MRA and could lead to a doubtful diagnosis. Therefore, we suggest that contamination should be prevented by using aseptic techniques as well as the MRA in those fibroblast cultures that cannot be discarded.

  19. Mycoplasma hyorhinis and Mycoplasma fermentans induce cell apoptosis and changes in gene expression profiles of 32D cells.

    PubMed

    Liu, Wenbin; Shou, Chengchao

    2011-01-01

    Infection of mycoplasmas has been linked to various human diseases including arthritis, pneumonia, infertility and cancer. While Mycoplasma hyorhinis and Mycoplasma fermentans have been detected in gastric adenocarcinomas, the mechanisms underlyine the pathogenesis are unknown. In this study, cell growth kinetics, Hoechst 33258 staining, DNA ladder assays, Western blotting analysis and cDNA microarray assays were performed to investigate the roles of M. hyorhinis and M. fermentans during infection of mammalian cells. Our data demonstrated that these mycoplasmas inhibid the growth of immortalised cell lines (32D and COS-7) ane tumor cell lines (HeLa and AGS). In addition, the infection of the 32D cell line with M. hyorhinis and M. fermentans induced compression of the nucleus, degradation of the cell genome and dysregulation of the expression of genes related to proliferation, apoptosis, tumorigenesis, signaling pathway and metabolism. Apoptosis related proteins Bcl-2, Bid and p53 were down-regulated, Fas was up-regulated and Bax was dysregulated in mycoplasma-infected 32D cells. Together, our data demonstrated that infection of mycoplasmas inhibitd cele growts through modification of gene expression profiles and post-translation modification of proliferation and apoptosis related proteins. PMID:22446603

  20. Cytoskeletal elements in the bacterium Mycoplasma pneumoniae

    NASA Astrophysics Data System (ADS)

    Hegermann, Jan; Herrmann, Richard; Mayer, Frank

    2002-09-01

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

  1. Mycoplasma mycoides, from "mycoides Small Colony" to "capri". A microevolutionary perspective

    PubMed Central

    2011-01-01

    Background The Mycoplasma mycoides cluster consists of five species or subspecies that are ruminant pathogens. One subspecies, Mycoplasma mycoides subspecies mycoides Small Colony (MmmSC), is the causative agent of contagious bovine pleuropneumonia. Its very close relative, Mycoplasma mycoides subsp. capri (Mmc), is a more ubiquitous pathogen in small ruminants causing mastitis, arthritis, keratitis, pneumonia and septicaemia and is also found as saprophyte in the ear canal. To understand the genetics underlying these phenotypic differences, we compared the MmmSC PG1 type strain genome, which was already available, with the genome of an Mmc field strain (95010) that was sequenced in this study. We also compared the 95010 genome with the recently published genome of another Mmc strain (GM12) to evaluate Mmc strain diversity. Results The MmmSC PG1 genome is 1,212 kbp and that of Mmc 95010 is ca. 58 kbp shorter. Most of the sequences present in PG1 but not 95010 are highly repeated Insertion Sequences (three types of IS) and large duplicated DNA fragments. The 95010 genome contains five types of IS, present in fewer copies than in PG1, and two copies of an integrative conjugative element. These mobile genetic elements have played a key role in genome plasticity, leading to inversions of large DNA fragments. Comparison of the two genomes suggested a marked decay of the PG1 genome that seems to be correlated with a greater number of IS. The repertoire of gene families encoding surface proteins is smaller in PG1. Several genes involved in polysaccharide metabolism and protein degradation are also absent from, or degraded in, PG1. Conclusions The genome of MmmSC PG1 is larger than that of Mmc 95010, its very close relative, but has less coding capacity. This is the result of large genetic rearrangements due to mobile elements that have also led to marked gene decay. This is consistent with a non-adaptative genomic complexity theory, allowing duplications or pseudogenes to

  2. Prevalence of Urogenital Mycoplasmas Among Men with NGU in Upper Silesia, Poland. Preliminary Study.

    PubMed

    Ekiel, Alicja; Aptekorz, Małgorzata; Kłuciński, Piotr; Smolec, Dominika; Wiechuła, Barbara; Jóźwiak, Jarosław; Martirosian, Gayane

    2016-01-01

    The prevalence of urogenital mycoplasmas in men with NGU in Upper Silesia (Poland) was studied. Mycoplasmas were detected in 36.7% men (Ureaplasma parvum and Mycoplasma genitalium were found in 30% and 16.7% respectively). Urealyticum urealyticum was not detected. We suggest including M. genitalium in the diagnostic scheme for nongonococcal urethritis (NGU). PMID:27281999

  3. Molecular characterisation of Mycoplasma hyorhinis isolated from pigs using pulsed-field gel electrophoresis and 16S rRNA sequencing.

    PubMed

    Yamaguti, Maurício; Oliveira, Rosângela C; Marques, Lucas M; Buzinhani, Melissa; Buim, Marcos R; Neto, Renata L; Guimarães, Ana Márcia S; Timenetsky, Jorge

    2015-01-01

    Economic loss in pig breeding is common due to respiratory disorders, and Mycoplasma hyopneumoniae and Mycoplasma hyorhinis, namely, are the most common infectious agents. The aim of this study is to recover these mollicutes and detect their genotypic variations by pulsed-field gel electrophoresis (PFGE) and sequencing the 16 s rRNA gene. One hundred and twenty-six swabs from tonsil and nasal mucus of pigs with respiratory disorders were analysed. A total of 78 lungs were sampled, as well as two trachea and two tonsils obtained from animals with respiratory disorder. A total of 59 isolates were obtained: 1 (1.70 per cent) of M hyopneumoniae, 2 (3.40 per cent) of Mycoplasma flocculare and 56 (94.90 per cent) of M hyorhinis. The PFGE for M hyorhinis showed 10 profiles with enzyme AvaI and 9 profiles with XhoI. A low polymorphism of the 16sRNS gene was detected in M hyorhinis isolates compared with the type strain in the GenBank. M hyorhinis isolates of different herds showed a large heterogenicity with enzymes AvaI and XhoI. The sequencing of the 16S rRNA gene allowed for analysing the interspecific and intraspecific variations of isolated mycoplasmas.

  4. Molecular characterisation of Mycoplasma hyorhinis isolated from pigs using pulsed-field gel electrophoresis and 16S rRNA sequencing

    PubMed Central

    Yamaguti, Maurício; Oliveira, Rosângela C; Marques, Lucas M; Buzinhani, Melissa; Buim, Marcos R; Neto, Renata L; Guimarães, Ana Márcia S; Timenetsky, Jorge

    2015-01-01

    Economic loss in pig breeding is common due to respiratory disorders, and Mycoplasma hyopneumoniae and Mycoplasma hyorhinis, namely, are the most common infectious agents. The aim of this study is to recover these mollicutes and detect their genotypic variations by pulsed-field gel electrophoresis (PFGE) and sequencing the 16 s rRNA gene. One hundred and twenty-six swabs from tonsil and nasal mucus of pigs with respiratory disorders were analysed. A total of 78 lungs were sampled, as well as two trachea and two tonsils obtained from animals with respiratory disorder. A total of 59 isolates were obtained: 1 (1.70 per cent) of M hyopneumoniae, 2 (3.40 per cent) of Mycoplasma flocculare and 56 (94.90 per cent) of M hyorhinis. The PFGE for M hyorhinis showed 10 profiles with enzyme AvaI and 9 profiles with XhoI. A low polymorphism of the 16sRNS gene was detected in M hyorhinis isolates compared with the type strain in the GenBank. M hyorhinis isolates of different herds showed a large heterogenicity with enzymes AvaI and XhoI. The sequencing of the 16S rRNA gene allowed for analysing the interspecific and intraspecific variations of isolated mycoplasmas. PMID:26688737

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

    PubMed

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

    2013-05-01

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

  6. Phenotypic and genotypic characterisation of Blastocystis hominis isolates implicates subtype 3 as a subtype with pathogenic potential.

    PubMed

    Tan, T C; Suresh, K G; Smith, H V

    2008-12-01

    Despite frequent reports on the presence of Blastocystis hominis in human intestinal tract, its pathogenicity remains a matter of intense debate. These discrepancies may be due to the varying pathogenic potential or virulence of the isolates studied. The present study represents the first to investigate both phenotypic and genotypic characteristics of B. hominis obtained from symptomatic and asymptomatic individuals. Symptomatic isolates had a significantly greater size range and lower growth rate in Jones' medium than asymptomatic isolates. The parasite cells of symptomatic isolates exhibited rougher surface topography and greater binding affinity to Canavalia ensiformis (ConA) and Helix pomatia (HPA). The present study also identifies further phenotypic characteristics, which aided in differentiating the pathogenic forms from the non-pathogenic forms of B. hominis. Blastocystis subtype 3 was found to be correlated well with the disease. PMID:18795333

  7. Serological and molecular survey of sheep infected with Mycoplasma ovipneumoniae in Xinjiang, China.

    PubMed

    Cheng, Chen; Jun, Qiao; Qingling, Meng; Zhengxiang, Hu; Yu, Ma; Xuepeng, Cai; Zibing, Cheng; Jinsheng, Zhang; Zaichao, Zhang; Kuojun, Cai; Chuangfu, Chen

    2015-12-01

    Mycoplasma pneumonia is one of the most important infectious diseases that threaten sheep production. In order to investigate the epidemic status of Mycoplasma ovipneumoniae infection in sheep, indirect hemagglutination assay was used to analyze 1679 serum samples collected from four different breeds of sheep (Kazak sheep, Hu sheep, Merino sheep, and Duolang sheep) in six regions in Xinjiang between 2012 and 2014. One thousand one hundred sixty-nine sheep nasal swabs and 180 lungs were PCR analyzed. The results showed that the average positive rates of the serum samples were 17.75 %. The positive rates were between 9.76 and 30.61 % in the four breeds. Among them, the Hu sheep had a significantly higher rate than other breeds (P < 0.05). The average positive rates of nasal swabs and lungs were 10.18 and 28.89 %, respectively. Based on the phylogenetic trees of 16S RNA gene, the isolates were closest to those strains isolated from inland areas of China, indicating that these epidemic isolates came from the trans-province introductions. Our survey suggests that quarantine is necessary for sheep imported from inland, and effective immunization should be implemented in sheep susceptible to M. ovipneumoniae in Xinjiang, China.

  8. A replicating plasmid-based vector for GFP expression in Mycoplasma hyopneumoniae.

    PubMed

    Ishag, H Z A; Liu, M J; Yang, R S; Xiong, Q Y; Feng, Z X; Shao, G Q

    2016-01-01

    Mycoplasma hyopneumoniae (M. hyopneumoniae) causes porcine enzootic pneumonia (PEP) that significantly affects the pig industry worldwide. Despite the availability of the whole genome sequence, studies on the pathogenesis of this organism have been limited due to the lack of a genetic manipulation system. Therefore, the aim of the current study was to generate a general GFP reporter vector based on a replicating plasmid. Here, we describe the feasibility of GFP reporter expression in M. hyopneumoniae (strain 168L) controlled by the p97 gene promoter of this mycoplasma. An expression plasmid (pMD18-TOgfp) containing the p97 gene promoter, and origin of replication (oriC) of M. hyopneumoniae, tetracycline resistant marker (tetM), and GFP was constructed and used to transform competent M. hyopneumoniae cells. We observed green fluorescence in M. hyopneumoniae transformants under fluorescence microscopy, which indicates that there was expression of the GFP reporter that was driven by the p97 gene promoter. Additionally, an electroporation method for M. hyopneumoniae with an efficiency of approximately 1 x 10(-6) transformants/μg plasmid DNA was optimized and is described herein. In conclusion, our data demonstrate the susceptibility of M. hyopneumoniae to genetic manipulation whereby foreign genes are expressed. This work may encourage the development of genetic tools to manipulate the genome of M. hyopneumoniae for functional genomic analyses. PMID:27173288

  9. GFP as a marker for transient gene transfer and expression in Mycoplasma hyorhinis.

    PubMed

    Ishag, Hassan Z A; Liu, Maojun; Yang, Ruosong; Xiong, Qiyan; Feng, Zhixin; Shao, Guoqing

    2016-01-01

    Mycoplasma hyorhinis (M. hyorhinis) is an opportunistic pathogen of pigs and has been shown to transform cell cultures, which has increased the interest of researchers. The green florescence proteins (GFP) gene of Aquorea victoria, proved to be a vital marker to identify transformed cells in mixed populations. Use of GFP to observe gene transfer and expression in M. hyorhinis (strain HUB-1) has not been described. We have constructed a pMD18-O/MHRgfp plasmid containing the p97 gene promoter, origin of replication, tetracycline resistance marker and GFP gene controlled by the p97 gene promoter. The plasmid transformed into M. hyorhinis with a frequency of ~4 × 10(-3) cfu/µg plasmid DNA and could be detected by PCR amplification of the GFP gene from the total DNA of the transformant mycoplasmas. Analysis of a single clone grown on KM2-Agar containing tetracycline, showed a green fluorescence color. Conclusively, this report suggests the usefulness of GFP to monitor transient gene transfer and expression in M. hyorhinis, eventually minimizing screening procedures for gene transfer and expression. PMID:27386255

  10. Antimicrobial susceptibility of Mycoplasma bovis isolates from veal calves and dairy cattle in the Netherlands.

    PubMed

    Heuvelink, Annet; Reugebrink, Constance; Mars, Jet

    2016-06-30

    Control of Mycoplasma bovis infections depends on good husbandry practices and antibiotic treatment. To allow more prudent use of antimicrobial drugs, there is a need for information on the susceptibility profile of this pathogen. The objective of the present study was to analyse the in vitro antimicrobial susceptibility of clinical M. bovis isolates in the Netherlands. The collection comprised 95 bovine isolates, originating from lungs (n=56), mastitis milk (n=27), and synovial fluid (n=12), collected between 2008 and 2014. Minimal inhibitory concentrations (MICs) were assessed by broth microdilution, both by using in-house prepared MIC plates and by using commercially available MIC plates. For each antimicrobial agent, the range of MIC results, the MIC50, and MIC90 values were calculated. M. bovis strains recently isolated in the Netherlands appeared to be characterized by relatively high MIC values for antimicrobial agents that, until now, have been recommended by the Dutch Association of Veterinarians for treating pneumonia caused by Mycoplasma species. Fluoroquinolones appeared to be the most efficacious in inhibiting M. bovis growth, followed by tulathromycin and oxytetracycline. The highest MIC values were obtained for erythromycin, tilmicosin, and tylosin. Future studies should be done on determining M. bovis specific clinical breakpoints, standardization of methods to determine MIC values as well as molecular studies on detection of antimicrobial resistance mechanisms of M. bovis isolates to develop PCR assays for determining resistance.

  11. Antimicrobial susceptibility of Mycoplasma bovis isolates from veal calves and dairy cattle in the Netherlands.

    PubMed

    Heuvelink, Annet; Reugebrink, Constance; Mars, Jet

    2016-06-30

    Control of Mycoplasma bovis infections depends on good husbandry practices and antibiotic treatment. To allow more prudent use of antimicrobial drugs, there is a need for information on the susceptibility profile of this pathogen. The objective of the present study was to analyse the in vitro antimicrobial susceptibility of clinical M. bovis isolates in the Netherlands. The collection comprised 95 bovine isolates, originating from lungs (n=56), mastitis milk (n=27), and synovial fluid (n=12), collected between 2008 and 2014. Minimal inhibitory concentrations (MICs) were assessed by broth microdilution, both by using in-house prepared MIC plates and by using commercially available MIC plates. For each antimicrobial agent, the range of MIC results, the MIC50, and MIC90 values were calculated. M. bovis strains recently isolated in the Netherlands appeared to be characterized by relatively high MIC values for antimicrobial agents that, until now, have been recommended by the Dutch Association of Veterinarians for treating pneumonia caused by Mycoplasma species. Fluoroquinolones appeared to be the most efficacious in inhibiting M. bovis growth, followed by tulathromycin and oxytetracycline. The highest MIC values were obtained for erythromycin, tilmicosin, and tylosin. Future studies should be done on determining M. bovis specific clinical breakpoints, standardization of methods to determine MIC values as well as molecular studies on detection of antimicrobial resistance mechanisms of M. bovis isolates to develop PCR assays for determining resistance. PMID:27259820

  12. 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

  13. Antibiotic Susceptibility of Biofilm Cells and Molecular Characterisation of Staphylococcus hominis Isolates from Blood

    PubMed Central

    Mendoza-Olazarán, Soraya; Morfín-Otero, Rayo; Villarreal-Treviño, Licet; Rodríguez-Noriega, Eduardo; Llaca-Díaz, Jorge; Camacho-Ortiz, Adrián; González, Gloria M.; Casillas-Vega, Néstor; Garza-González, Elvira

    2015-01-01

    Objectives We aimed to characterise the staphylococcal cassette chromosome mec (SCCmec) type, genetic relatedness, biofilm formation and composition, icaADBC genes detection, icaD expression, and antibiotic susceptibility of planktonic and biofilm cells of Staphylococcus hominis isolates from blood. Methods The study included 67 S. hominis blood isolates. Methicillin resistance was evaluated with the cefoxitin disk test. mecA gene and SCCmec were detected by multiplex PCR. Genetic relatedness was determined by pulsed-field gel electrophoresis. Biofilm formation and composition were evaluated by staining with crystal violet and by detachment assay, respectively; and the biofilm index (BI) was determined. Detection and expression of icaADBC genes were performed by multiplex PCR and real-time PCR, respectively. Antibiotic susceptibilities of planktonic cells (minimum inhibitory concentration, MIC) and biofilm cells (minimum biofilm eradication concentration, MBEC) were determined by the broth dilution method. Results Eighty-five percent (57/67) of isolates were methicillin resistant and mecA positive. Of the mecA-positive isolates, 66.7% (38/57) carried a new putative SCCmec type. Four clones were detected, with two to five isolates each. Among all isolates, 91% (61/67) were categorised as strong biofilm producers. Biofilm biomass composition was heterogeneous (polysaccharides, proteins and DNA). All isolates presented the icaD gene, and 6.66% (1/15) isolates expressed icaD. This isolate presented the five genes of ica operon. Higher BI and MBEC values than the MIC values were observed for amikacin, vancomycin, linezolid, oxacillin, ciprofloxacin, and chloramphenicol. Conclusions S. hominis isolates were highly resistant to methicillin and other antimicrobials. Most of the detected SCCmec types were different than those described for S. aureus. Isolates indicated low clonality. The results indicate that S. hominis is a strong biofilm producer with an extracellular

  14. Sarcocystis rommeli, n. sp. (Apicomplexa: Sarcocystidae) from Cattle (Bos taurus) and its Differentiation from Sarcocystis hominis.

    PubMed

    Dubey, Jitender P; Moré, Gastón; van Wilpe, Erna; Calero-Bernal, Rafael; Verma, Shiv K; Schares, Gereon

    2016-01-01

    Cattle (Bos taurus) are intermediate hosts for three named species of Sarcocystis, S. cruzi, S. hirsuta, and S. hominis. Recently, a fourth species was identified and named S. sinensis. However, S. sinensis originally named a species of Sarcocystis in water buffalo (Bubalus bubalis) in China. Based on unverifiable evidence, it was suggested that the same parasite infects cattle. In addition, S. sinensis was recently declared as nomen nudum because its naming violated the rules of International Code of Zoological Nomenclature. Thus, the fourth species using cattle as an intermediate host does not have a valid name. Here, we propose a new name, Sarcocystis rommeli for the S. sinensis-like parasite from cattle in Argentina, and differentiate it ultrastructurally from S. hominis sarcocysts from experimentally infected cattle. Sarcocystis rommeli sarcocysts were microscopic with a 5-μm-thick wall with slender villar protrusions (Vp); the Vp were up to 5 μm long, up to 0.5 μm wide, and of uneven thickness, often bent at an angle. The ground substance layer (Gs) was up to 0.8 μm thick and smooth. Vesicular structures were seen at the base of the Vp. The bradyzoites were 10-12 μm long. Sarcocystis hominis sarcocysts had Vp that were often upright, up to 7.5 μm long, and up to 1.8 μm wide; the Gs was up to 2 μm thick and without vesicles. Its sarcocyst wall was up to 5.6 μm thick, the vp were bent at an angle, up to 5.8 μm long, the Gs was up to 2 μm thick, but without vesicles seen in S. rommeli. Beef containing sarcocysts of S. rommeli was not orally infectious for two human volunteers and a red fox (Vulpes vulpes). The Sarcocystis described here is molecularly different from S. cruzi, S. hirsuta, and S. hominis based on 18S rRNA and cox1 gene sequences.

  15. 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

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

    PubMed

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

    2006-01-01

    Respiratory disease in sheep and goats is a major problem in Jordan and is often associated with Mycoplasma species. Without effective vaccines, control is mainly by chemotherapy, but the uncontrolled use of antimicrobials has led to concerns about the potential development of antimicrobial resistance. The in vitro effect of chloramphenicol, florfenicol, enrofloxacin, tylosin, erythromycin and oxytetracycline was determined against 32 isolates of Mycoplasma species-M. mycoides subsp. mycoides LC (6), M. capricolum subsp. capricolum (8) and M. putrefaciens (18), all isolated from either nasal swabs or milk, from sheep and goats in different regions of Jordan. The antimicrobial susceptibility showed some Mycoplasma species-specific differences, with M. capricolum subsp. capricolum being more susceptible to tylosin and erythromycin. Chloramphenicol and florfenicol were the least effective for all three Mycoplasma species. No trends or significant differences in antimicrobial susceptibilities were observed between sheep and goat isolates, between milk or nasal swab isolates, or between isolates from different regions of Jordan. Some isolates of M. capricolum subsp. capricolum and M. putrefaciens showed higher MIC levels with oxytetracycline, as did two isolates of M. mycoides subsp. mycoides LC with tylosin, possibly indicating signs of development of antimicrobial resistance. PMID:17405622

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

    PubMed

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

    2016-06-01

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

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

    PubMed Central

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

    2015-01-01

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

  19. Substituted Pyrrolo[2,3-d]pyrimidines as Cryptosporidium hominis Thymidylate Synthase Inhibitors

    PubMed Central

    Kumar, Vidya P.; Frey, Kathleen M.; Wang, Yiqiang; Jain, Hitesh K.; Gangjee, Aleem; Anderson, Karen S.

    2013-01-01

    Cryptosporidiosis, a gastrointestinal disease caused by a protozoan Cryptosporidium hominis is often fatal in immunocompromised individuals. There is little clinical data to show that the existing treatment by nitazoxanide and paromomycin is effective in immunocompromised individuals1, 2. Thymidylate synthase (TS) and dihydrofolate reductase (DHFR) are essential enzymes in the folate biosynthesis pathway and are well established as drug targets in cancer and malaria. A novel series of classical antifolates, 2-amino-4-oxo-5-substituted pyrrolo[2,3-d]pyrimidines have been evaluated as Cryptosporidium hominis thymidylate synthase (ChTS) inhibitors. Crystal structure in complex with the most potent compound, a 2’-chlorophenyl with a sulfur bridge with a Ki of 8.83 ± 0.67 nM is discussed in terms of several Van de Waals, hydrophobic and hydrogen bond interactions with the protein residues and the substrate analog 5-fluorodeoxyuridine monophosphate. Of these interactions, two interactions with the non-conserved residues (A287 and S290) offer an opportunity to develop ChTS specific inhibitors. Compound 6 serves as a lead compound for analog design and its crystal structure provides clues for the design of ChTS specific inhibitors. PMID:23927969

  20. Genotypic Characterization of Cryptosporidium hominis from Water Samples in São Paulo, Brazil

    PubMed Central

    Araújo, Ronalda S.; Dropa, Milena; Fernandes, Licia N.; Carvalho, Terezinha T.; Sato, Maria Inês Z.; Soares, Rodrigo M.; Matté, Glavur R.; Matté, Maria Helena

    2011-01-01

    The protozoan parasite Cryptosporidium has emerged as one of the most important water contaminants, causing waterborne outbreaks of diarrheal diseases worldwide. The small size of oocysts under the microscope and the possibility of changes in characteristics of oocysts, mainly in environmental samples, make the taxonomy of the genus difficult if morphologic characteristics are considered. This limitation encouraged the application of molecular methods to identify this microorganism. The aim of this study was to detect and identify by nested-polymerase chain reaction oocysts of Cryptosporidium present in water samples in the state of São Paulo, Brazil. Water samples were concentrated through a membrane filter, DNA was extracted by using a standard technique, and both amplification reactions used forward and reverse oligonucleotides that were complementary to Cryptosporidium 18S ribosomal RNA gene sequences. Thirty water samples from different sites of collection in the state of São Paulo were evaluated. Cryptosporidium oocysts were detected in 30% of the samples. By genoptyping, C. hominis and Cryptosporidium sp. were identified in recreational water and C. meleagridis was identified in surface water samples. This is the first report of C. hominis in environmental samples in Brazil. Although identification of Cryptosporidium is still a difficult task, molecular methods are essential for specific identification and are a helpful tool to aid to understand the epidemiology of this parasite in Brazil. PMID:22049036

  1. Molecular cloning of HSP70 in Mycoplasma ovipneumoniae and comparison with that of other mycoplasmas.

    PubMed

    Li, M; Ma, C J; Liu, X M; Zhao, D; Xu, Q C; Wang, Y J

    2011-05-10

    Mycoplasma ovipneumoniae, a bacterial species that specifically affects ovine and goat, is the cause of ovine infectious pleuropneumonia. We cloned, sequenced and analyzed heat shock protein 70 (HSP70) (dnaK) gene of M. ovipneumoniae. The full length open reading frame of the M. ovipneumoniae HSP70 gene consists of 1812 nucleotides, with a G+C content of 34.16%, encoding 604 amino acids. Comparative analysis with the HSP70 sequences of 15 Mycoplasma species revealed 59 to 87% DNA sequence identity, with an amino acid sequence identity range of 58 to 94%. M. ovipneumoniae and M. hyopneumoniae shared the highest DNA and amino acid sequence identity (87 and 94%, respectively). Based on phylogenetic analysis, both the DNA and amino acid identities of M. ovipneumoniae with other mycoplasmal HSP70 were correlated with the degree of relationship between the species. The C-terminus of the HSP70 was cloned into a bacterial expression vector and expressed in Escherichia coli cells. The recombinant C-terminal portion of HSP70 protein strongly reacted with convalescent sera from M. ovipneumoniae-infected sheep, based on an immunoblotting assay. This indicates that HSP70 is immunogenic in a natural M. ovipneumoniae infection and may be a relevant antigen for vaccine development.

  2. Rapid Detection of Contagious Bovine Pleuropneumonia by a Mycoplasma mycoides subsp. mycoides SC Capsular Polysaccharide-Specific Antigen Detection Latex Agglutination Test

    PubMed Central

    March, John B.; Kerr, Karen; Lema, Benedict

    2003-01-01

    A latex agglutination test (LAT) has been developed for the diagnosis of contagious bovine pleuropneumonia (CBPP). The latex microspheres were coated with MmmSC polyclonal immunoglobulin G antiserum and detected MmmSC antigen in the serum of cattle infected with CBPP and in growth medium containing MmmSC. The specific antigen recognizsed by this test appeared to be the capsular polysaccharide (CPS). The LAT recognized all 23 strains of MmmSC examined in this study, with a sensitivity level of 2 ng of CPS, or the equivalent of 5 × 103 CFU, in a reaction volume of 0.03 ml. Therefore, rapid identification of MmmSC cultures should be possible. Agglutination was also observed with the related goat pathogens and “Mycoplasma mycoides” cluster members Mycoplasma mycoides subsp. mycoides large colony biotype (four of six strains positive) and Mycoplasma mycoides subsp. capri (three of six strains positive), in agreement with the suggestion that these latter two mycoplasmas may in fact represent a single species (although collectively exhibiting two capsular serotypes). Comparisons in diagnosis with the complement fixation test (CFT) were made by using African field sera from CBPP-infected cattle. After 2 (or 3) min of incubation, the test detected 55% (or 61%) of CFT-positive sera and 29% (or 40%) of CFT-negative sera, with an overall correlation in diagnosis of 62% (or 61%). The rates for false-positive diagnoses made by using “known” CBPP-negative sera from the United Kingdom were 3 or 13% after 2 or 3 min of incubation, respectively. The data agree with previous findings that some CBPP CFT-negative misdiagnoses may occur due to “antibody eclipsing” by excess circulating antigen. The LAT combines low cost and high specificity with ease of application in the field, without the need for any specialist training or equipment. PMID:12626448

  3. Gliding Motility of Mycoplasma mobile on Uniform Oligosaccharides

    PubMed Central

    Kasai, Taishi; Hamaguchi, Tasuku

    2015-01-01

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

  4. Erosive polyarthritis associated with Mycoplasma gateae in a cat.

    PubMed

    Zeugswetter, Florian; Hittmair, Katharina M; de Arespacochaga, Abigail G; Shibly, Sarina; Spergser, Joachim

    2007-06-01

    Erosive polyarthritis was diagnosed in an 11-month-old neutered male Egyptian Mau-cross cat with concurrent glucocorticoid-responsive dermatitis. Clinical signs, synovial fluid analysis, serological tests and radiographic appearance could not differentiate between immune-mediated and infective arthritis. Mycoplasma gateae was isolated by strictly anaerobic culture of the synovial fluid. Treatment with Enrofloxacin led to a rapid improvement of the cat's condition. Two months later the cat was euthanased because of severe glomerulonephritis and direct Coombs' test positive anaemia, possibly caused by mycoplasma infection. M gateae could not be isolated at post-mortem examination. PMID:17175189

  5. Mycoplasma genitalium: An Emerging Sexually Transmitted Infection

    PubMed Central

    Munoz, Jessian L.

    2016-01-01

    Mycoplasma genitalium has been recognized as a cause of male urethritis, and there is now evidence suggesting that it causes cervicitis and pelvic inflammatory disease in women. M. genitalium is a slow growing organism, and, with the advent of nucleic acid amplification test (NAAT), more studies are being performed, and knowledge about the pathogenicity of this organism elucidated. With NAAT detection, treatment modalities have been studied, and the next challenge is to determine the most effective antimicrobial therapy. Doxycycline, the first-line antibiotic for urethritis, is largely ineffective in the treatment of M. genitalium and furthermore, resistance to macrolide has also emerged. The most effective drug is Moxifloxacin although there are emerging reports of resistance to it in various parts of the world. This paper not only highlights the current research and knowledge, but also reviews the diversity of health implications on the health of men and women infected with M. genitalium. Alternate antibiotics and the impact of M. genitalium on infertility are areas that require more studies as we continue to research into this microorganism. PMID:27034904

  6. Mechanisms of volume regulation in Mycoplasma gallisepticum

    SciTech Connect

    Linker, C.S.

    1987-01-01

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

  7. Mycoplasma genitalium: Should We Treat and How?

    PubMed Central

    Broad, Jennifer M.; Golden, Matthew R.

    2011-01-01

    Mycoplasma genitalium is associated with acute and chronic urethritis in men. Existing data on infection in women are limited and inconsistent but suggest that M. genitalium is associated with urethritis, cervicitis, pelvic inflammatory disease, and possibly female infertility. Data are inconclusive regarding the role of M. genitalium in adverse pregnancy outcomes and ectopic pregnancy. Available data suggest that azithromycin is superior to doxycycline in treating M. genitalium infection. However, azithromycin-resistant infections have been reported in 3 continents, and the proportion of azithromycin-resistant M. genitalium infection is unknown. Moxifloxacin is the only drug that currently seems to uniformly eradicate M. genitalium. Detection of M. genitalium is hampered by the absence of a commercially available diagnostic test. Persons with persistent pelvic inflammatory disease or clinically significant persistent urethritis or cervicitis should be tested for M. genitalium, if possible. Infected persons who have not previously received azithromycin should receive that drug. Persons in whom azithromycin therapy fails should be treated with moxifloxicin. PMID:22080266

  8. Cell volume regulation in Mycoplasma gallisepticum.

    PubMed

    Linker, C; Wilson, T H

    1985-09-01

    Mycoplasma gallisepticum cells incubated in 250 mM NaCl solutions in the absence of glucose showed a progressive fall in intracellular ATP concentration over a period of 2 to 3 h. When the ATP level fell below 40 microM the cell began to swell and become progressively permeable to [14C]inulin and leak intracellular protein and nucleotides. The addition of nondiffusable substances such as MgSO4 or disaccharides prevented swelling, suggesting that NaCl (and water) entry was due to Gibbs-Donnan forces. The addition of glucose after the initiation of cell swelling increased intracellular ATP, induced cell shrinkage, and prevented the release of intracellular components. The ATPase inhibitor dicyclohexylcarbodiimide, which collapsed the chemical and electrical components of the proton motive force, caused rapid cell swelling in the presence of glucose (and high intracellular ATP levels). Extracellular impermeable solutes such as MgSO4 and disaccharides prevented swelling of dicyclohexylcarbodiimide-treated cells incubated in NaCl. It was postulated that Na+ that diffused into the cell was extruded by an electrogenic Na+-H+ exchange (antiport) energized by the proton motive force established by the dicyclohexylcarbodiimide-sensitive H+-ATPase.

  9. Serological study of Mycoplasma pneumoniae infections.

    PubMed

    Srifuengfung, Somporn; Techachaiwiwat, Wanida; Dhiraputra, Chertsak

    2004-08-01

    Mycoplasma pneumoniae antibody was determined in 811 sera of different patients admitted to Siriraj Hospital with respiratory tract infection from July 1, 2000 to August 31, 2003 by agglutination with gelatin particle agglutination test kit (SERODIA-MYCO II, Fujirebio Inc. Japan) in microtiter plates. Three hundred and three sera were positive (37.36%). The five most positive titer were found in patients 5-9 yr (40.26%), followed by patients 1-4 yr (24.75%), 10-14 yr (19.80%), 30-39 yr (5.28%) and 20-29 yr (3.96%). The positive titers ranged from 40 to > 20,480. Female:male ratio in positive patients was approximately the same (1.19:1). High titers (> or = 320) were found in 146 out of 303 patients (48.18%). The infection was mostly found in children aged 5-9 yr. Detection of antibody to M. pneumoniae infection showed that 37.36% of patients who were suspected of having atypical bacterial pneumonia were positive.

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

    PubMed Central

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

    2016-01-01

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

  11. 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.

  12. Prevalence and Genotype Characterization of Blastocystis hominis Among the Baghmalek People in Southwestern Iran in 2013 - 2014

    PubMed Central

    Khoshnood, Saleh; Rafiei, Abdollah; Saki, Jasem; Alizadeh, Kobra

    2015-01-01

    Background: Blastocystis hominis is a common globally distributed parasite. The prevalence of this parasite has been shown to vary among different countries. Molecular studies have also shown that there is a high level of genetic diversity among Blastocystis spp. isolated from humans and animals. Extensive information on parasitic genotypes will aid in devising more effective strategies for the identification and potential control of these pathogenic parasites. Objectives: This study aimed to gain information on the prevalence and abundance of Blastocystis subtypes in Iran. Materials and Methods: Over a period of 3 months, 1,410 stool samples were collected and examined by microscopy. Samples found to be positive for B. hominis were concentrated and phylogenetic analysis was subsequently performed. A questionnaire was completed by all study participants. Results: Blastocystis hominis was found to have a prevalence of 3.33% in the study population. There was no significant association of Blastocystis infection with age (P = 0.3) or gender (P = 0.57). The Blastocystis subtypes (ST) identified in this study were ST3, ST4, ST5, and ST7 with the most prevalent being ST4 (40.9%). Conclusions: The prevalence of B. hominis in the study area was lower than that reported for most developed countries, and unlike in other countries in the Middle East, ST4 was the most prevalent subtype. PMID:26587213

  13. Warble? What’s a Warble? A recap of the human bot fly, Dermatobia hominis (L. Jr. 1781)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The human bot fly, Dermatobia hominis (Linnaeus Jr., 1781) is a major pest of livestock in Mexico, Central and South America. Myiasis caused by the larvae result in economic losses due to hide damage and reductions in weight gain and milk production. They have a broad host range which includes wildl...

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

    PubMed Central

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

    2012-01-01

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

  15. Comparison of in vitro activity of danofloxacin, florfenicol, oxytetracycline, spectinomycin and tilmicosin against Mycoplasma mycoides subspecies mycoides small colony type.

    PubMed

    Ayling, R D; Baker, S E; Nicholas, R A; Peek, M L; Simon, A J

    2000-02-26

    Minimum inhibitory concentrations (MIC) and minimum mycoplasmacidal concentrations (MMC) of the antimicrobials danofloxacin, florfenicol, oxytetracycline, spectinomycin and tilmicosin were determined in vitro for 20 isolates of Mycoplasma mycoides subspecies mycoides small colony type (MmmSC), the causative agent of contagious bovine pleuropneumonia (CBPP). The majority of strains were most susceptible to tilmicosin, followed by danofloxacin, oxytetracycline, florfenicol and spectinomycin with MIC50 values of 0.015, 0.25, 0.5, 1 and 8 microg/ml, and MMC50 values of 0.06, 0.5, 8, 8 and 16 microg/ml, respectively. However, tilmicosin had poor mycoplasmacidal activity against two recent strains from Portugal. There was no evidence of resistance to danofloxacin in any of the strains. PMID:10737293

  16. In vitro pharmacodynamics of gamithromycin against Mycoplasma mycoides subspecies mycoides Small Colony.

    PubMed

    Mitchell, John D; Goh, Shan; McKellar, Quintin A; McKeever, Declan J

    2013-09-01

    Mycoplasma mycoides mycoides Small Colony (MmmSC) is the causative agent of contagious bovine pleuropneumonia (CBPP), which is responsible for major economic losses in sub-Saharan Africa. Current control relies on live attenuated vaccines, which are of limited efficacy, and antimicrobials are now being assessed as an alternative or adjunct to vaccination. The objective of this study was to determine the in vitro effector kinetics of the macrolide antimicrobial, gamithromycin, against MmmSC in artificial medium and adult bovine serum. Furthermore, it was determined if any differences in gamithromycin activity between these two matrices were mirrored by the older macrolides, tylosin and tilmicosin. Minimum inhibitory concentrations (MICs) for gamithromycin, tylosin and tilmicosin against MmmSC strains B237 and Tan8 were determined in artificial medium and serum. Time-kill curves were constructed at concentrations corresponding to multiples of the MIC for all three macrolides in artificial medium and for gamithromycin in serum. Data were fitted to sigmoid Emax models. Post-antibiotic effects (PAE) were established by exposing strain B237 to antimicrobials at 10× MIC for 1h and monitoring mycoplasma growth thereafter. MICs for gamithromycin, tylosin and tilmicosin were 64-, 8- and 64-fold lower, respectively, in serum than in artificial medium at an inoculum size of 10(6)cfu/mL B237. A similar pattern emerged for Tan8. All three antimicrobials were mycoplasmastatic with maximum effects of -0.44, -0.32 and -0.49log10(cfu/mL) units for gamithromycin, tylosin and tilmicosin, respectively, against B237 in artificial medium. Tylosin and tilmicosin elicited longer PAEs than gamithromycin. In conclusion, gamithromycin, tylosin and tilmicosin all demonstrated in vitro efficacy against MmmSC and represent potential candidates for clinical studies to assess their therapeutic effect against CBPP. PMID:23810743

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

    PubMed

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

    2015-09-01

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

  18. In vitro pharmacodynamics of gamithromycin against Mycoplasma mycoides subspecies mycoides Small Colony.

    PubMed

    Mitchell, John D; Goh, Shan; McKellar, Quintin A; McKeever, Declan J

    2013-09-01

    Mycoplasma mycoides mycoides Small Colony (MmmSC) is the causative agent of contagious bovine pleuropneumonia (CBPP), which is responsible for major economic losses in sub-Saharan Africa. Current control relies on live attenuated vaccines, which are of limited efficacy, and antimicrobials are now being assessed as an alternative or adjunct to vaccination. The objective of this study was to determine the in vitro effector kinetics of the macrolide antimicrobial, gamithromycin, against MmmSC in artificial medium and adult bovine serum. Furthermore, it was determined if any differences in gamithromycin activity between these two matrices were mirrored by the older macrolides, tylosin and tilmicosin. Minimum inhibitory concentrations (MICs) for gamithromycin, tylosin and tilmicosin against MmmSC strains B237 and Tan8 were determined in artificial medium and serum. Time-kill curves were constructed at concentrations corresponding to multiples of the MIC for all three macrolides in artificial medium and for gamithromycin in serum. Data were fitted to sigmoid Emax models. Post-antibiotic effects (PAE) were established by exposing strain B237 to antimicrobials at 10× MIC for 1h and monitoring mycoplasma growth thereafter. MICs for gamithromycin, tylosin and tilmicosin were 64-, 8- and 64-fold lower, respectively, in serum than in artificial medium at an inoculum size of 10(6)cfu/mL B237. A similar pattern emerged for Tan8. All three antimicrobials were mycoplasmastatic with maximum effects of -0.44, -0.32 and -0.49log10(cfu/mL) units for gamithromycin, tylosin and tilmicosin, respectively, against B237 in artificial medium. Tylosin and tilmicosin elicited longer PAEs than gamithromycin. In conclusion, gamithromycin, tylosin and tilmicosin all demonstrated in vitro efficacy against MmmSC and represent potential candidates for clinical studies to assess their therapeutic effect against CBPP.

  19. Chronic non-progressive pneumonia of sheep in New Zealand - a review of the role of Mycoplasma ovipneumoniae.

    PubMed

    Alley, M R; Ionas, G; Clarke, J K

    1999-10-01

    Chronic non-progressive pneumonia (CNP) is a common disease which affects lambs in New Zealand during late summer and autumn. Mycoplasma ovipneumoniae can be recovered from a high proportion of lesions but it is also present in some normal lungs. Bacteria, especially Pasteurella haemolytica, can also be recovered from more than half the lungs of affected animals. Isolates of M. ovipneumoniae are genetically heterogeneous, as demonstrated by examination of their DNA or total cellular proteins, and are serologically heterogeneous as shown by metabolic inhibition tests. The number of strains present in New Zealand is large and several distinguishable strains can be recovered from each affected lung. Mycoplasma ovipneumoniae has pathogenic potential as indicated by its ability to produce hydrogen peroxide, cause ciliostasis and by its possession of a capsule. Chronic non-progressive pneumonia can be transmitted consistently to over 50% of lambs by inoculation of pooled pneumonic lung homogenate and transmission can be suppressed by broad spectrum antibiotics. In contrast, penicillin does not prevent the development of lesions but diminishes their severity. Pooled lung homogenate treated with digitonin, which inactivates mycoplasmas, has failed to transmit CNP. Pure cultures of M. ovipneumoniae produce only mild lesions in some animals, whereas inoculation with pooled lung homogenate (from which no viruses were isolated) containing mixed strains of M. ovipneumoniae and free from bacteria, is more effective in producing lesions. Research work to date suggests that CNP may be initiated by colonisation of the lung by M. ovipneumoniae which causes ciliostasis and elicits an exudate allowing colonisation of the lungs by bacteria especially M. haemolytica and by other strains of M. ovipneumoniae. The immune response to the initial strain of M. ovipneumoniae may inhibit its replication but would be less effective in inhibiting heterologous strains of the organism allowing

  20. A study of the heterogeneity of isolates of Mycoplasma ovipneumoniae from sheep in New Zealand.

    PubMed

    Ionas, G; Norman, N G; Clarke, J K; Marshall, R B

    1991-11-01

    To investigate the heterogeneity of Mycoplasma ovipneumoniae, sixty isolates from three sheep on each of twenty farms were examined by restriction endonuclease analysis (REA) and SDS-PAGE. All were found to be different except for three isolates obtained from one farm. The protein and REA patterns of individual isolates were both highly reproducible and remained unchanged following long term passage (approximately 400 generations) in vitro. No plasmids were detected in the twelve strains which were examined and when two isolates were co-cultured in vitro, no genetic interchange, as judged by changes in REA patterns were detected. Since the heterogeneity of M. ovipneumoniae when examined by SDS-PAGE is too great to allow groups to be recognised, it could be advantageous for this purpose if only surface proteins were compared. As a preliminary step to this end we have identified several surface proteins of M. ovipneumoniae and found that some are common to all strains, one surface protein was shared by five of the eight strains examined and another was unique to one strain. This approach has the potential to allow the recognition of grouping of M. ovipneumoniae isolates.

  1. The immunodominant 90-kilodalton protein is localized on the terminal tip structure of Mycoplasma pneumoniae.

    PubMed Central

    Franzoso, G; Hu, P C; Meloni, G A; Barile, M F

    1993-01-01

    Immunoblot analysis of convalescent-phase sera of experimentally infected chimpanzees or monoclonal antibodies (MAbs) specific to the 90- and 40-kDa proteins of Mycoplasma pneumoniae indicated that both proteins were present in cytadsorbing, pathogenic strains PI-1428, M129, and FH but absent in noncytadsorbing, nonpathogenic strain M129-B176. Adsorption of convalescent-phase chimpanzee sera with virulent strain PI-1428 removed reactivity, whereas adsorption with avirulent strain M129-B176 did not remove reactivity to these two proteins. By using proteolysis and specific MAbs, we demonstrated that the 90- and 40-kDa proteins were surface exposed. Immunoelectron microscopy employing specific MAbs showed that the 90-kDa protein is localized on the terminal tip attachment apparatus. However, the MAb specific for the 40-kDa protein failed to indicate a similar localization. Nevertheless, these data, taken together, indicate that the immunodominant 90- and 40-kDa proteins are surface exposed, are localized on the terminal tip apparatus, and might be involved in the attachment mechanism. Images PMID:8454358

  2. Chlamydiae and Mycoplasma infections in pulmonary MALT lymphoma

    PubMed Central

    Chanudet, E; Adam, P; Nicholson, A G; Wotherspoon, A C; Ranaldi, R; Goteri, G; Pileri, S A; Ye, H; Müller-Hermelink, H K; Du, M-Q

    2007-01-01

    Chlamydia pneumoniae, Chlamydia trachomatis and Chlamydia psittaci were detected at low frequencies (<20%) among 69 pulmonary mucosa-associated lymphoid tissue (MALT) lymphomas, 30 other lymphoproliferative disorders (LPD) and 44 non-LPD. The incidence of individual Chlamydiae was generally higher in MALT lymphoma than non-LPD, although not reaching statistical significance. Mycoplasma pneumoniae DNA was not detected. PMID:17876330

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

    PubMed Central

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

    2015-01-01

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

  4. Bloodstream Infection Due to Mycoplasma arginini in an Immunocompromised Patient

    PubMed Central

    Watanabe, Mayumi; Goto, Miki; Hasegawa, Yuichi

    2012-01-01

    Mycoplasma arginini, an organism usually recovered from mammals, was isolated from the blood of a febrile patient with advanced non-Hodgkin lymphoma. The patient's condition improved without administration of antimycoplasmal drugs. Simulation of blood culture showed that automated blood culture instruments may fail to detect the organism. PMID:22785195

  5. Unravelling the Transcriptome Profile of the Swine Respiratory Tract Mycoplasmas

    PubMed Central

    Siqueira, Franciele Maboni; Gerber, Alexandra Lehmkuhl; Guedes, Rafael Lucas Muniz; Almeida, Luiz Gonzaga; Schrank, Irene Silveira; Vasconcelos, Ana Tereza Ribeiro; Zaha, Arnaldo

    2014-01-01

    The swine respiratory ciliary epithelium is mainly colonized by Mycoplasma hyopneumoniae, Mycoplasma flocculare and Mycoplasma hyorhinis. While colonization by M. flocculare is virtually asymptomatic, M. hyopneumoniae and M. hyorhinis infections may cause respiratory disease. Information regarding transcript structure and gene abundance provides valuable insight into gene function and regulation, which has not yet been analyzed on a genome-wide scale in these Mycoplasma species. In this study, we report the construction of transcriptome maps for M. hyopneumoniae, M. flocculare and M. hyorhinis, which represent data for conducting comparative studies on the transcriptional repertory. For each species, three cDNA libraries were generated, yielding averages of 415,265, 695,313 and 93,578 reads for M. hyopneumoniae, M. flocculare and M. hyorhinis, respectively, with an average read length of 274 bp. The reads mapping showed that 92%, 98% and 96% of the predicted genes were transcribed in the M. hyopneumoniae, M. flocculare and M. hyorhinis genomes, respectively. Moreover, we showed that the majority of the genes are co-expressed, confirming the previously predicted transcription units. Finally, our data defined the RNA populations in detail, with the map transcript boundaries and transcription unit structures on a genome-wide scale. PMID:25333523

  6. Interaction of Mycoplasma dispar with bovine alveolar macrophages.

    PubMed Central

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

    1992-01-01

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

  7. Mycoplasma bovis: An emerging pathogen of ranched bison

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

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

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

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

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

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

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

  12. 21 CFR 866.3375 - Mycoplasma spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Mycoplasma spp. serological reagents. 866.3375 Section 866.3375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3375...

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

    PubMed

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

    2016-10-01

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

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

    PubMed

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

    2016-10-01

    Bovine mastitis caused by mycoplasmas, in particular Mycoplasma bovis, is a major problem for milk production and animal welfare in large dairy herds in the USA and a serious, although sporadic, disease in Europe and the Middle East. It causes severe damage to the udder of cattle and is largely untreatable by chemotherapy. Mycoplasma mastitis has a distinct epidemiology and a unique set of risk factors, the most important of which is large herd size. The disease is often self-limiting, disappearing within months of outbreaks, sometimes without deliberate intervention. Improved molecular diagnostic tests are leading to more rapid detection of mycoplasmas. Typing tests, such as multi-locus sequence typing, can help trace the source of outbreaks. An approach to successful control is proposed, which involves regular monitoring and rapid segregation or culling of infected cows. Serious consideration should be given by owners of healthy dairy herds to the purchase of M. bovis-free replacements. Increased cases of disease could occur in Europe and Israel if the trend for larger dairy herds continues. PMID:27687942

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

    PubMed

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

    2009-12-01

    The isolation of Mycoplasma hyosynoviae from a piglet with severe pneumonia is described. This is the first report of M. hyosynoviae isolation in the country. The lung sample where the isolation was made was severely consolidated, suppurative and pleurisy. The pathogenicity of the M. hyosynoviae isolated has yet to be determined. PMID:20237449

  16. 21 CFR 610.30 - Test for Mycoplasma.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 7 2014-04-01 2014-04-01 false Test for Mycoplasma. 610.30 Section 610.30 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) BIOLOGICS... Methylene Blue-Azure dye or an equivalent staining procedure is used, no less than a one square cm. plug...

  17. [Mycoplasma pneumoniae: a cause of febrile hemolytic anemia in travelers].

    PubMed

    Ficko, C; Andriamanantena, D; Flateau, F; Mangouka, L; Soler, C; Carmoi, T; Rapp, C

    2012-01-01

    Mycoplasma pneumoniae can cause varied hematologic manifestations that are frequently associated with lower respiratory tract infections. Acute febrile hemolysis without respiratory symptoms is quite rare. We describe the case of a 25-year-old man, admitted for acute fever with hemolysis, after returning from Djibouti. M. pneumoniae infection was proved by serological testing. A favorable outcome followed macrolide treatment. PMID:23352983

  18. 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

  19. Species-Specific Monoclonal Antibodies to Escherichia coli-Expressed p36 Cytosolic Protein of Mycoplasma hyopneumoniae

    PubMed Central

    Caron, J.; Sawyer, N.; Moumen, B. Ben Abdel; Bouh, K. Cheikh Saad; Dea, S.

    2000-01-01

    The p36 protein of Mycoplasma hyopneumoniae is a cytosolic protein carrying species-specific antigenic determinants. Based on the genomic sequence of the reference strain ATCC 25934, primers were designed for PCR amplification of the p36-encoding gene (948 bp). These primers were shown to be specific to M. hyopneumoniae since no DNA amplicons could be obtained with other mycoplasma species and pathogenic bacteria that commonly colonize the porcine respiratory tract. The amplified p36 gene was subcloned into the pGEX-4T-1 vector to be expressed in Escherichia coli as a fusion protein with glutathione S-transferase (GST). The GST-p36 recombinant fusion protein was purified by affinity chromatography and cut by thrombin, and the enriched p36 protein was used to immunize female BALB/c mice for the production of anti-p36 monoclonal antibodies (MAbs). The polypeptide specificity of the nine MAbs obtained was confirmed by Western immunoblotting with cell lysates prepared from the homologous strain. Cross-reactivity studies of the anti-p36 MAbs towards two other M. hyopneumoniae reference strains (ATCC 25095 and J strains) and Quebec field strains that had been isolated in culture suggested that these anti-p36 MAbs were directed against a highly conserved epitope, or closely located epitopes, of the p36 protein. No reactivity was demonstrated against other mycoplasma species tested. Clinical signs and lesions suggestive of enzootic pneumonia were reproduced in specific-pathogen-free pigs infected experimentally with a virulent Quebec field strain (IAF-DM9827) of M. hyopneumoniae. The bacteria could be recovered from lung homogenates of pigs that were killed after the 3-week observation period by both PCR and cultivation procedures. Furthermore, the anti-p36 MAbs permitted effective detection by indirect immunofluorescence of M. hyopneumoniae in frozen lung sections from experimentally infected pigs. However, attempts to use the recombinant p36 protein as an antigen in an

  20. The pathogenicity of Mycoplasma ovipneumoniae and Mycoplasma arginini in ovine and caprine tracheal organ cultures.

    PubMed

    Jones, G E; Keir, W A; Gilmour, J S

    1985-10-01

    The effects of M. ovipneumoniae and M. arginini on tracheal organ cultures prepared from a neonatal kid and a foetal lamb were studied. Both organisms were isolated from the cultures throughout the 14 days of observation. M. ovipneumoniae produced ciliostasis and loss of cilia, confirmed by scanning electron microscope (SEM), after 4 days. These effects were sudden and profound in lamb explants, and gradual and less pronounced in kid explants. Clusters of organisms attached to epithelial surfaces and in association with cilia were visible by SEM. M. arginini also induced ciliostasis and cilia loss in both kid and lamb explants, but onset was more rapid, at 2 days, and there was evident recovery after day 6, with apparent regeneration of cilia. No clearly recognizable mycoplasmas were observed by SEM in M. arginini-infected explants.

  1. Cryptosporidium hominis genotypes involved in increased incidence and clusters of cases, Navarra, Spain, 2012.

    PubMed

    Fuentes, I; Martín, C; Beristain, X; Mazón, A; Saugar, J M; Blanco, A; García Cenoz, M; Valle-Cristia, M; Ezpeleta, C; Castilla, J

    2015-04-01

    SUMMARY Two clusters of confirmed cryptosporidiosis infections were detected in Navarra, Spain, in the summer of 2012, in the context of an increased incidence in the region. Molecular subtyping of Cryptosporidium hominis determined that one cluster, occurring in an urban area, was due to the predominant circulating subtype IbA10G2R2 and the other cluster, with cases occurring in a rural area, was due to a rare subtype IaA18R3. No single exposure was associated with infection, although exposure to certain children's pools was reported by a majority of patients interviewed in each cluster. Genotyping tools were useful in the investigation and could aid investigation of cryptosporidiosis outbreaks in Spain in the future.

  2. Staphylococcus hominis native mitral valve bacterial endocarditis (SBE) in a patient with hypertrophic obstructive cardiomyopathy.

    PubMed

    Cunha, Burke A; Esrick, Michael D; Larusso, Melissa

    2007-01-01

    There are several species of coagulase-negative Staphylococci (CoNS) that are part of the normal skin flora and are relatively noninvasive/low virulence organisms. CoNS are important pathogens in patients with prosthetic devices and are the most common pathogen associated with prosthetic valve endocarditis. CoNS native valve infective endocarditis (IE) is rare. Patients with hypertrophic obstructive cardiomyopathy and an outflow pressure gradient greater than 30 mm Hg are predisposed to IE. There has been only one reported case of non-mitral valve IE due to CoNS in a patient with hypertrophic obstructive cardiomyopathy. To the best of our knowledge, we report the first case of Staphylococcal hominis mitral valve endocarditis in a patient with hypertrophic obstructive cardiomyopathy.

  3. Complete circular DNA in the mitochondria-like organelles of Blastocystis hominis.

    PubMed

    Wawrzyniak, Ivan; Roussel, Michaël; Diogon, Marie; Couloux, Arnaud; Texier, Catherine; Tan, Kevin S W; Vivarès, Christian P; Delbac, Frédéric; Wincker, Patrick; El Alaoui, Hicham

    2008-10-01

    Blastocystis hominis is an anaerobic parasite of the human intestinal tract belonging to the Stramenopile group. Using genome sequencing project data, we describe here the complete sequence of a 29,270-bp circular DNA molecule that presents mitochondrial features (such as oxidative phosphorylation complex I subunits) but lacks complexes III, IV and V. Transmission electron microscopy analyses reveal that this molecule, as well as mitochondrial (NADH:ubiquinone oxidoreductase subunit 7 (NAD7), beta-succinyl-CoA synthetase (beta-SCS)) and hydrogenosomal (pyruvate ferredoxin oxido-reductase (PFOR), iron-hydrogenase) proteins, are located within double-membrane surrounded-compartments known as mitochondria-like organelles (MLOs). As there is no evidence for hydrogen production by this organism, we suggest that MLOs are more likely anaerobic mitochondria. PMID:18694756

  4. What are mycoplasmas - The relationship of tempo and mode in bacterial evolution

    NASA Technical Reports Server (NTRS)

    Woese, C. R.; Stackebrand, E.; Ludwig, W.

    1985-01-01

    In phenotype the mycoplasmas are very different from ordinary bacteria. However, genotypically (i.e., phylogenetically) they are not. On the basis of ribosomal RNA homologies the mycoplasmas belong with the clostridia, and indeed have specific clostridial relatives. Mycoplasmas are, however, unlike almost all other bacteria in the evolutionary characteristics of their ribosomal RNAs. These RNAs contain relatively few of the highly conserved oligonucleotide sequences characteristic of normal eubacterial ribosomal RNAs. This is interpreted to be a reflection of an elevated mutation rate in mycoplasma lines of descent. A general consequence of this would be that the variation associated with a mycoplasma population is augmented both in number and kind, which in turn would lead to an unusual evolutionary course, one unique in all respects. Mycoplasmas, then, are actually tachytelic bacteria. The unusual evolutionary characteristics of their ribosomal RNAs are the imprints of their rapid evolution.

  5. What are mycoplasmas: the relationship of tempo and mode in bacterial evolution

    NASA Technical Reports Server (NTRS)

    Woese, C. R.; Stackebrandt, E.; Ludwig, W.

    1984-01-01

    In phenotype the mycoplasmas are very different from ordinary bacteria. However, genotypically (i.e., phylogenetically) they are not. On the basis of ribosomal RNA homologies the mycoplasmas belong with the clostridia, and indeed have specific clostridial relatives. Mycoplasmas are, however, unlike almost all other bacteria in the evolutionary characteristics of their ribosomal RNAs. These RNAs contain relatively few of the highly conserved oligonucleotide sequences characteristic of normal eubacterial ribosomal RNAs. This is interpreted to be a reflection of an elevated mutation rate in mycoplasma lines of descent. A general consequence of this would be that the variation associated with a mycoplasma population is augmented both in number and kind, which in turn would lead to an unusual evolutionary course, one unique in all respects. Mycoplasmas, then, are actually tachytelic bacteria. The unusual evolutionary characteristics of their ribosomal RNAs are the imprints of their rapid evolution.

  6. Novel hemotropic Mycoplasma species in white-tailed deer (Odocoileus virginianus).

    PubMed

    Maggi, Ricardo G; Chitwood, M Colter; Kennedy-Stoskopf, Suzanne; DePerno, Christopher S

    2013-12-01

    Globally, hemotropic Mycoplasma spp. are emerging or re-emerging zoonotic pathogens that affect livestock, wildlife, companion animals, and humans, potentially causing serious and economically important disease problems. Little is known about hemotropic Mycoplasma spp. prevalence, host-specificity, or route of transmission in most species, including wildlife. DNA amplification by PCR targeting the 16SrRNA and the RNaseP genes was used to establish the presence and prevalence of hemotropic Mycoplasma spp. in a white-tailed deer (O. virginianus) population in eastern North Carolina. Sixty-five deer (89%) tested positive for hemotropic Mycoplasma spp. where sequence analysis of the 16SsRNA and the RNaseP genes indicated the presence of at least three distinct species. This study represents the first detection of three distinct hemotropic Mycoplasma species in white-tailed deer and the first report of two novel hemotropic Mycoplasma species.

  7. Recovery of Mycoplasma spp. from the Reproductive Tract of the Mare during the Estrous Cycle

    PubMed Central

    Bermudez, Victor; Miller, Richard; Johnson, Walter; Rosendal, Soren; Ruhnke, Louise

    1987-01-01

    The sites in the genital tract from which mycoplasmas could be recovered at various stages of the estrous cycle were studied in five Standardbred mares naturally infected with Mycoplasma. Mycoplasma equigenitalium and Mycoplasma subdolum were most frequently isolated from the clitoral fossa as compared to the vagina, cervix, and uterus. The lowest isolation prevalence was observed in the uterus. The recovery of Mycoplasma spp. from the clitoral fossa did not differ at any stage of the estrous cycle; however, recovery from the vagina, cervix, and uterus was variable during the cycle and more organisms were recovered on the day of ovulation than at any other time. From these results it was concluded that the clitoral fossa is the most likely “ecological niche” for Mycoplasma spp. in the mare. Ureaplasmas were not isolated. ImagesFigure 1.Figure 2. PMID:17422844

  8. Prevalence of Mycoplasma gallisepticum and Mycoplasma synoviae in commercial poultry, racing pigeons and wild birds in Belgium.

    PubMed

    Michiels, Tinne; Welby, Sarah; Vanrobaeys, Mia; Quinet, Christian; Rouffaer, Lieze; Lens, Luc; Martel, An; Butaye, Patrick

    2016-01-01

    Mycoplasma gallisepticum is the most important pathogenic avian Mycoplasma species and causes chronic respiratory disease in poultry. In addition, the prevalence of Mycoplasma synoviae is of increasing concern in several EU member states. We investigated the prevalence of M. gallisepticum in commercial poultry (5220 layers, 1224 broilers and 1020 meat turkeys), 56 racing pigeons and 890 wild birds (Order Anseriformes, Galliformes, Pelecaniformes, Accipitriformes, Gruiformes, Charadriiformes, Columbiformes, Strigiformes, Falconiformes and Passeriformes). Broilers and wild birds were also evaluated for Mycoplasma synoviae. Dependent on the bird lifespan and the nature of the sample, different diagnostic tests were used including the rapid plate agglutination test, enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction and real-time polymerase chain reaction. A low prevalence of M. gallisepticum was found in both layers (0.9%; 95% CI: 0.7-1.2%) and broilers (2.7%; 95% CI: 1.9-3.8%) possibly due to reduced vertical transmission by breeder farms, which are under official surveillance. None of the samples from turkeys or racing pigeons tested positive. In wild birds, we found five birds were positive (1.7%; 95% CI: 0.7-3.9%): one wood pigeon, two grey herons, one mallard and one Eurasian magpie. For M. synoviae a high prevalence was found in broilers (12.9%: 95% CI: 11.1-14.9%). Four samples collected by hunters gave a positive result for M. synoviae (4%: 95% CI: 1.6-9.8%): one carrion crow and three wood pigeons. In addition, 12 house sparrows were found to be positive (3%; 95% CI: 1.7-5.2%). Wild birds probably play a limited role as a reservoir but we cannot exclude a possible impact on transmission of Mycoplasmas.

  9. Prevalence of Mycoplasma gallisepticum and Mycoplasma synoviae in commercial poultry, racing pigeons and wild birds in Belgium.

    PubMed

    Michiels, Tinne; Welby, Sarah; Vanrobaeys, Mia; Quinet, Christian; Rouffaer, Lieze; Lens, Luc; Martel, An; Butaye, Patrick

    2016-01-01

    Mycoplasma gallisepticum is the most important pathogenic avian Mycoplasma species and causes chronic respiratory disease in poultry. In addition, the prevalence of Mycoplasma synoviae is of increasing concern in several EU member states. We investigated the prevalence of M. gallisepticum in commercial poultry (5220 layers, 1224 broilers and 1020 meat turkeys), 56 racing pigeons and 890 wild birds (Order Anseriformes, Galliformes, Pelecaniformes, Accipitriformes, Gruiformes, Charadriiformes, Columbiformes, Strigiformes, Falconiformes and Passeriformes). Broilers and wild birds were also evaluated for Mycoplasma synoviae. Dependent on the bird lifespan and the nature of the sample, different diagnostic tests were used including the rapid plate agglutination test, enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction and real-time polymerase chain reaction. A low prevalence of M. gallisepticum was found in both layers (0.9%; 95% CI: 0.7-1.2%) and broilers (2.7%; 95% CI: 1.9-3.8%) possibly due to reduced vertical transmission by breeder farms, which are under official surveillance. None of the samples from turkeys or racing pigeons tested positive. In wild birds, we found five birds were positive (1.7%; 95% CI: 0.7-3.9%): one wood pigeon, two grey herons, one mallard and one Eurasian magpie. For M. synoviae a high prevalence was found in broilers (12.9%: 95% CI: 11.1-14.9%). Four samples collected by hunters gave a positive result for M. synoviae (4%: 95% CI: 1.6-9.8%): one carrion crow and three wood pigeons. In addition, 12 house sparrows were found to be positive (3%; 95% CI: 1.7-5.2%). Wild birds probably play a limited role as a reservoir but we cannot exclude a possible impact on transmission of Mycoplasmas. PMID:26814376

  10. The recovery of a Mycoplasma from Citellus richardsonii richardsonii (ground squirrel).

    PubMed Central

    Langford, E V

    1977-01-01

    Swabs from the upper respiratory tract, external genitalia and the eyes and portions of the lungs, spleen, kidney, liver and uteri of two ground squirrels were cultured for mycoplasma. The upper respiratory tracts of both animals were positive for mycoplasma as were the lungs, liver and spleen of one of the animals. Preliminary serological studies, growth inhibition test against 38 known antisera, growth precipitation against 15 antisera and fluorescent antibody technique with eight conjugates have, with the exception of a minor precipitin reaction against Mycoplasma bovirhinis, all been negative. The isolates are believed to be representative of either one or more new mycoplasma species. PMID:861842

  11. Comparison of the prevalence of Mycoplasma species in dogs with and without respiratory disease.

    PubMed

    Schulz, Bianka S; Raufeisen, Katharina; Weber, Karin; Laberke, Siija; Hartmann, Katrin

    2015-01-01

    Aim of the study was to investigate the prevalence of Mycoplasma species in dogs with and without signs of respiratory disease. Bronchoalveolarlavage fluid (BALF) and pharyngeal swabs were collected from 29 dogs with respiratory diseases (RD) and 16 dogs without signs of RD that were euthanised because of other diseases. Samples were tested for Mycoplasma species by PCR and culture, and sequencing was performed in Mycoplasma species-positive BALF samples. Pharyngeal swabs were positive for Mycoplasma species by PCR in 91.7% of dogs with RD and 86.7% of dogs without signs of RD (p = 1.000); BALF samples were PCR-positive in 37.9% of dogs with RD and 18.8% of dogs without signs of RD (p = 0.194) Mycoplasmo culture of BALF was positive in 28.6% of dogs with RD and in 18.8% without signs of RD (p = 0.730). When culture and PCR were compared, there was no significant difference in the detection rate of Mycoplasma species (p = 0.658) Sequencing detected different Mycoplasma species. Out of these, however, Mycoplasma cynos was isolated from four dogs with RD. There is no significant difference in the prevalence of Mycoplasma species between dogs with RD and dogs without evidence of RD; however, Mycoplasma cynos seems to be associated with respiratory disease.

  12. [Serological analysis of coagulase-negative staphylococci: study of characteristic agglutinogens in type strains of the nine species individualized by Schleifer and Kloos (author's transl)].

    PubMed

    Pillet, J; Orta, B

    1977-01-01

    The occurrence of characteristic agglutinogens has been searched for in the type strains of the following species: Staphylococcus xylosus, S. cohnii, S. epidermidis, S. capitis, S. saprophyticus, S. warneri, S. haemolyticus, S. hominis and S. simulans. Serotyping of these nine strains and study of their antisera have been carried out with formalin-treated and autoclaved bacteria. It has been shown that the characteristic agglutinogens of S. epidermidis and S. haemolyticus were respectively identical to the agglutinogens previously described in the coagulase-negative type strain 52.186 and in the S. aureus type strain 18. Characteristic agglutinogens have been found in each of the other seven type strains. Agglutinogens found in S. xylosus and S. saprophyticus are thermolabile, the corresponding absorbed sera reacting only with the formalin-treated homologous strain. Concerning S. cohnii, S. capitis, S. hominis and S. simulans, their absorbed sera reacting both with formalin-treated and autoclaved homologous strains, the observed reactions can be accounted either by one thermostable agglutinogen or by two characteristic agglutinogens, one thermostable, the other thermolabile. In S. warneri, only one thermostable agglutinogen has been characterized, the specificity of the reaction observed between the absorbed S. warneri antiserum and the homologous formalin-treated strain having to be confirmed. The use for serotyping of absorbed sera prepared against the type strains of the species S. xylosus, S. cohnii, S. capitis, S. saprophyticus, S. warneri, S. hominis and S. simulans should permit to improve the individualization of coagulase-negative staphylococci. PMID:610502

  13. The History of Mycoplasma pneumoniae Pneumonia.

    PubMed

    Saraya, Takeshi

    2016-01-01

    transmission experiments resulted in a lapse of 20 years before acceptance of the Eaton agent as Mycoplasma pneumoniae. This review describes the history of M. pneumoniae pneumonia with a special focus on the recognition between the 1930 and 1960s of the Eaton agent as the infectious cause. PMID:27047477

  14. Development and evaluation of a multi-locus sequence typing scheme for Mycoplasma synoviae.

    PubMed

    Dijkman, R; Feberwee, A; Landman, W J M

    2016-08-01

    Reproducible molecular Mycoplasma synoviae typing techniques with sufficient discriminatory power may help to expand knowledge on its epidemiology and contribute to the improvement of control and eradication programmes of this mycoplasma species. The present study describes the development and validation of a novel multi-locus sequence typing (MLST) scheme for M. synoviae. Thirteen M. synoviae isolates originating from different poultry categories, farms and lesions, were subjected to whole genome sequencing. Their sequences were compared to that of M. synoviae reference strain MS53. A high number of single nucleotide polymorphisms (SNPs) indicating considerable genetic diversity were identified. SNPs were present in over 40 putative target genes for MLST of which five target genes were selected (nanA, uvrA, lepA, ruvB and ugpA) for the MLST scheme. This scheme was evaluated analysing 209 M. synoviae samples from different countries, categories of poultry, farms and lesions. Eleven clonal clusters and 76 different sequence types (STs) were obtained. Clustering occurred following geographical origin, supporting the hypothesis of regional population evolution. M. synoviae samples obtained from epidemiologically linked outbreaks often harboured the same ST. In contrast, multiple M. synoviae lineages were found in samples originating from swollen joints or oviducts from hens that produce eggs with eggshell apex abnormalities indicating that further research is needed to identify the genetic factors of M. synoviae that may explain its variations in tissue tropism and disease inducing potential. Furthermore, MLST proved to have a higher discriminatory power compared to variable lipoprotein and haemagglutinin A typing, which generated 50 different genotypes on the same database. PMID:26926568

  15. Antimicrobial Susceptibility Patterns of Recent Cuban Mycoplasma genitalium Isolates Determined by a Modified Cell-Culture-Based Method

    PubMed Central

    Mondeja, Brian A.; Rodríguez, Nadia M.; Barroto, Brenda; Blanco, Orestes

    2016-01-01

    Isolation of Mycoplasma genitalium from clinical specimens remains difficult and few strains are available for antimicrobial susceptibility testing. We describe the antimicrobial susceptibility of M. genitalium strains grown in Vero cell culture with first- and second- line antibiotics, using a modified cell-culture-based method. Macrolide- and -fluoroquinolone resistance determinants were detected by sequencing of the 23S and parC genes, respectively. Seven strains were examined, including three new, genetically distinct M. genitalium strains isolated from endocervical and urethral swab specimens from Cuban patients together with four reference strains isolated from specimens collected from men in Denmark, Sweden and Australia. Azithromycin was the most active drug against two of the Cuban M. genitalium strains with MICs values of 0.008 mg/liter, however, one strain was macrolide resistant with an MIC of >8 mg/liter, and the A2059G resistant genotype. Ciprofloxacin was the least active antimicrobial drug and moxifloxacin was the most active fluoroquinolone against the new clinical strains, although an MIC of 1 mg/l was found for two strains. However, no relevant parC mutations were detected. MICs for tetracyclines were 0.5–4 mg/liter. Although the number of Cuban strains was low, the results suggest that a single-dose azithromycin treatment could be ineffective, and that a second-line treatment with moxifloxacin, should become an option in Cuba. To our knowledge, this is the first report of isolation and antibiotic susceptibility testing of M. genitalium strains from the Latin-American region, and the first detection of macrolide resistance in such strains. PMID:27679992

  16. A Novel SimpleProbe PCR Assay for Detection of Mutations in the 23S rRNA Gene Associated with Macrolide Resistance in Mycoplasma genitalium in Clinical Samples

    PubMed Central

    Lysvand, Hilde; Pukstad, Brita; Nordbø, Svein Arne

    2016-01-01

    Macrolide-resistant strains of Mycoplasma genitalium are an increasing problem throughout the world, and the implementation of a rapid and sensitive assay for mutation detection to guide treatment is needed. Macrolide-resistant strains have been shown to contain base substitutions in positions 2058 and 2059 (Escherichia coli numbering) in region V of the 23S rRNA gene. In this study, we present a SimpleProbe PCR followed by melting curve analysis to differentiate between macrolide-resistant mutants and wild types. The assay was performed on 159 Mycoplasma genitalium-positive samples, and the results were compared with DNA sequencing. We also looked at the prevalence of macrolide-resistant strains in a Norwegian population. Of 139 samples characterized successfully by sequencing, 54 (39%) were wild types and 85 (61%) were mutants, consisting of 59 (42%) A2059G, 24 (17%) A2058G, 1 (1%) A2058T, and 1 (1%) A2059C mutation. The melting curve analysis correctly differentiated between wild-type and mutant strains in all cases, but it could not identify the different mutant types. The SimpleProbe PCR proved to be a simple, rapid, and reliable method for the detection of macrolide-resistant isolates of Mycoplasma genitalium in a clinical setting. PMID:27487958

  17. Furuncular myiasis caused by the human bot-fly Dermatobia hominis in a domestic cat from Brazil.

    PubMed

    Verocai, Guilherme G; Fernandes, Julio I; Correia, Thais R; de Souza, Clarissa P; Melo, Raquel M P S; Scott, Fabio B

    2010-06-01

    This paper reports a case of furuncular myiasis caused by the human bot-fly Dermatobia hominis in a domestic cat from Brazil. A crossbred shorthaired female cat of approximately 3 years old, presented with three boil-like cutaneous lesions at the left cranioventral region of the neck. These were diagnosed as furuncular myiasis. The animal was sedated, and after shaving the fur, bot-fly larvae were removed from the lesion by digital compression. Afterwards, the wounds were treated with 10% iodine solution and also with wound-healing cream containing sulfanilamide, urea and beeswax. Maggots were identified as third-stage larvae of D hominis. Clinical case reports of human bot-fly myiasis in cats are relevant due to its scarce occurrence in feline veterinary practice in some countries.

  18. Phenotypic, Genotypic, and Antimicrobial Characteristics of Streptococcus halichoeri Isolates from Humans, Proposal To Rename Streptococcus halichoeri as Streptococcus halichoeri subsp. halichoeri, and Description of Streptococcus halichoeri subsp. hominis subsp. nov., a Bacterium Associated with Human Clinical Infections.

    PubMed

    Shewmaker, P L; Whitney, A M; Humrighouse, B W

    2016-03-01

    Phenotypic, genotypic, and antimicrobial characteristics of six phenotypically distinct human clinical isolates that most closely resembled the type strain of Streptococcus halichoeri isolated from a seal are presented. Sequencing of the 16S rRNA, rpoB, sodA, and recN genes; comparative whole-genome analysis; conventional biochemical and Rapid ID 32 Strep identification methods; and antimicrobial susceptibility testing were performed on the human isolates, the type strain of S. halichoeri, and type strains of closely related species. The six human clinical isolates were biochemically indistinguishable from each other and showed 100% 16S rRNA, rpoB, sodA, and recN gene sequence similarity. Comparative 16S rRNA gene sequencing analysis revealed 98.6% similarity to S. halichoeri CCUG 48324(T), 97.9% similarity to S. canis ATCC 43496(T), and 97.8% similarity to S. ictaluri ATCC BAA-1300(T). A 3,530-bp fragment of the rpoB gene was 98.8% similar to the S. halichoeri type strain, 84.6% to the S. canis type strain, and 83.8% to the S. ictaluri type strain. The S. halichoeri type strain and the human clinical isolates were susceptible to the antimicrobials tested based on CLSI guidelines for Streptococcus species viridans group with the exception of tetracycline and erythromycin. The human isolates were phenotypically distinct from the type strain isolated from a seal; comparative whole-genome sequence analysis confirmed that the human isolates were S. halichoeri. On the basis of these results, a novel subspecies, Streptococcus halichoeri subsp. hominis, is proposed for the human isolates and Streptococcus halichoeri subsp. halichoeri is proposed for the gray seal isolates. The type strain of the novel subspecies is SS1844(T) = CCUG 67100(T) = LMG 28801(T).

  19. Phenotypic, Genotypic, and Antimicrobial Characteristics of Streptococcus halichoeri Isolates from Humans, Proposal To Rename Streptococcus halichoeri as Streptococcus halichoeri subsp. halichoeri, and Description of Streptococcus halichoeri subsp. hominis subsp. nov., a Bacterium Associated with Human Clinical Infections

    PubMed Central

    Whitney, A. M.; Humrighouse, B. W.

    2016-01-01

    Phenotypic, genotypic, and antimicrobial characteristics of six phenotypically distinct human clinical isolates that most closely resembled the type strain of Streptococcus halichoeri isolated from a seal are presented. Sequencing of the 16S rRNA, rpoB, sodA, and recN genes; comparative whole-genome analysis; conventional biochemical and Rapid ID 32 Strep identification methods; and antimicrobial susceptibility testing were performed on the human isolates, the type strain of S. halichoeri, and type strains of closely related species. The six human clinical isolates were biochemically indistinguishable from each other and showed 100% 16S rRNA, rpoB, sodA, and recN gene sequence similarity. Comparative 16S rRNA gene sequencing analysis revealed 98.6% similarity to S. halichoeri CCUG 48324T, 97.9% similarity to S. canis ATCC 43496T, and 97.8% similarity to S. ictaluri ATCC BAA-1300T. A 3,530-bp fragment of the rpoB gene was 98.8% similar to the S. halichoeri type strain, 84.6% to the S. canis type strain, and 83.8% to the S. ictaluri type strain. The S. halichoeri type strain and the human clinical isolates were susceptible to the antimicrobials tested based on CLSI guidelines for Streptococcus species viridans group with the exception of tetracycline and erythromycin. The human isolates were phenotypically distinct from the type strain isolated from a seal; comparative whole-genome sequence analysis confirmed that the human isolates were S. halichoeri. On the basis of these results, a novel subspecies, Streptococcus halichoeri subsp. hominis, is proposed for the human isolates and Streptococcus halichoeri subsp. halichoeri is proposed for the gray seal isolates. The type strain of the novel subspecies is SS1844T = CCUG 67100T = LMG 28801T. PMID:26763962

  20. 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

    ... mycoplasma reactors by in vivo bio-assay (enrichment). 147.16 Section 147.16 Animals and Animal Products... the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment). This procedure has been shown... publications: (a) Bigland, C. H. and A. J. DaMassa, “A Bio-Assay for Mycoplasma Gallisepticum.” In:...

  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

    ... mycoplasma reactors by in vivo bio-assay (enrichment). 147.16 Section 147.16 Animals and Animal Products... the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment). This procedure has been shown... publications: (a) Bigland, C. H. and A. J. DaMassa, “A Bio-Assay for Mycoplasma Gallisepticum.” In:...

  2. 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

    ... mycoplasma reactors by in vivo bio-assay (enrichment). 147.16 Section 147.16 Animals and Animal Products... the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment). This procedure has been shown... publications: (a) Bigland, C. H. and A. J. DaMassa, “A Bio-Assay for Mycoplasma Gallisepticum.” In:...

  3. 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

    ... mycoplasma reactors by in vivo bio-assay (enrichment). 147.16 Section 147.16 Animals and Animal Products... the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment). This procedure has been shown... publications: (a) Bigland, C. H. and A. J. DaMassa, “A Bio-Assay for Mycoplasma Gallisepticum.” In:...

  4. 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

    ... mycoplasma reactors by in vivo bio-assay (enrichment). 147.16 Section 147.16 Animals and Animal Products... the evaluation of mycoplasma reactors by in vivo bio-assay (enrichment). This procedure has been shown... publications: (a) Bigland, C. H. and A. J. DaMassa, “A Bio-Assay for Mycoplasma Gallisepticum.” In:...

  5. Mycoplasmas in Australian fur seals (Arctocephalus pusillus doriferus): identification and association with abortion.

    PubMed

    Lynch, Michael; Taylor, Trevor K; Duignan, Pádraig J; Swingler, Jane; Marenda, Marc; Arnould, John P Y; Kirkwood, Roger

    2011-11-01

    Bacteria from the genus Mycoplasma are common inhabitants of the respiratory, gastrointestinal, and genital tracts of mammals. The understanding of the pathological significance of mycoplasmas in seals is poor, as few studies have utilized the specific culture techniques required to isolate these bacteria. The current study surveyed for the Mycoplasma species present in Australian fur seals (Arctocephalus pusillus doriferus) and investigated the association between infection and pathology. Mycoplasmas were found in the nasal cavities of 55/80 (69%) of apparently healthy individuals. Isolates from 18 individuals were investigated through 16S ribosomal RNA sequencing, and 3 species were identified: M. zalophi, M. phocae, and Mycoplasma sp. (GenBank no. EU714238.1), all of which had previously been isolated from Northern Hemisphere pinnipeds. In addition, mycoplasmas were isolated from the lungs of 4 out of 16 juveniles and 1 out of 5 adults sampled at necropsy. Isolates obtained were M. zalophi, Mycoplasma sp. EU714238.1, and M. phocicerebrale, but infection was not associated with lung pathology in these age classes. Inflammatory disease processes of the heart and/or lungs were present in 12 out of 32 (38%) aborted fetuses on microscopic examination. Predominant findings were interstitial pneumonia, pericarditis, and myocarditis. Mycoplasma phocicerebrale was isolated from the thymus of an aborted fetus, and 3 out of 11 (27%) fetuses with inflammatory heart or lung lesions were PCR-positive for Mycoplasma. In conclusion, several species of Mycoplasma are part of the normal flora of the nasal cavity of Australian fur seals, and some mycoplasmas may be associated with abortion in this species of seal. PMID:22362792

  6. DNA sequences identical to Pneumocystis carinii f. sp. carinii and Pneumocystis carinii f. sp. hominis in samples of air spora.

    PubMed Central

    Wakefield, A E

    1996-01-01

    Samples of ambient air collected with three different types of spore traps in a rural location were examined for the presence of Pneumocystis carinii by screening for P. carinii-specific DNA sequences by DNA amplification. Eleven spore trap samples were analyzed by nested PCR, using oligonucleotide primers designed for the gene encoding the mitochondrial large subunit rRNA of P. carinii f. sp. carinii and P. carinii f. sp. hominis. The samples were collected over a 3-year period during the months of May to September, with a range of sampling times from 9 to 240 h. One air sample from an animal facility housing P. carinii-infected rats was also examined. P. carinii-specific amplification products were obtained from samples from each of the spore traps. The amplification products from eight air samples were cloned and sequenced. The majority of the recombinants from each of these samples had sequences identical to those of P. carinii f. sp. carinii and P. carinii f. sp. hominis, and a number of clones had single-base differences. These data suggest that sequences identical to those of P. carinii f. sp. carinii and P. carinii f. sp. hominis can be detected in samples of air collected in a rural location and that P. carinii may be a component of the air spora of rural Oxfordshire. PMID:8784583

  7. Unusual Enterocytozoon bieneusi genotypes and Cryptosporidium hominis subtypes in HIV-infected patients on highly active antiretroviral therapy.

    PubMed

    Akinbo, Frederick O; Okaka, Christopher E; Omoregie, Richard; Adamu, Haileeyesus; Xiao, Lihua

    2013-07-01

    Human immunodeficiency virus (HIV)-infected persons are commonly infected with Cryptosporidium species and Enterocytozoon bieneusi in both developed and developing countries, particularly patients with CD4+ cell counts below 200 cells/μL; 285 HIV-infected patients on highly active antiretroviral therapy (HAART) were enrolled in this study, and both stool and blood specimens were collected from participants. The stool specimens were analyzed and typed for E. bieneusi and Cryptosporidium spp. by polymerase chain reaction (PCR) and DNA sequencing. CD4 count was analyzed using flow cytometry. E. bieneusi and Cryptosporidium were detected in 18 (6.3%) and 4 (1.4%) patients, respectively. The E. bieneusi detected mostly belonged to a new genotype group that, thus far, has only been found in a few humans: genotype Nig4 in 2 patients and two new genotypes related to Nig4 in 12 patients. The Cryptosporidium detected included C. hominis (two patients), C. parvum (one patient), and C. felis (one patient), with the two C. hominis infections belonging to an unusual subtype family. Additional studies are required to determine whether some E. bieneusi genotypes and C. hominis subtypes are more prevalent in HIV patients on HAART. PMID:23629938

  8. Mycoplasma testudineum in free-ranging desert tortoises, Gopherus agassizii

    USGS Publications Warehouse

    Jacobson, Elliott R.; Berry, Kristin H.

    2012-01-01

    We performed clinico-pathological evaluations of 11 wild Agassiz's desert tortoises (Gopherus agassizii) from a translocation project in the central Mojave Desert, California, USA. Group 1 consisted of nine tortoises that were selected primarily due to serologic status, indicating exposure to Mycoplasma testudineum (seven) or both M. agassizii and M. testudineum (two), and secondarily due to clinical signs of upper respiratory tract disease (URTD). Group 2 consisted of two tortoises that were antibody-negative for Mycoplasma and had no clinical signs of URTD, but did have other signs of illness. Of the Group 1 tortoises, M. testudineum, but not M. agassizii, was amplified by polymerase chain reaction and DNA fingerprinted from two tortoises. Using light microscopy, mild to severe pathologic changes were observed in one or more histologic sections of either one or both nasal cavities of each tortoise in Group 1. Our findings support a causal relationship between M. testudineum and URTD in desert tortoises.

  9. A change in the genetic code in Mycoplasma capricolum

    NASA Technical Reports Server (NTRS)

    Jukes, T. H.

    1985-01-01

    Mycoplasma capricolum was previously found to use UGA instead of UGG as its codon for tryptophan and to contain 75 percent A + T in its DNA. The codon change could have been due to mutational pressure to replace C + G by A + T, resulting in the replacement of UGA stop codons by UAA, change of the anticodon in tryptophan tRNA from CCA to UCA, and replacement of UGG tryptophan codons by UGA. None of these changes should have been deleterious.

  10. Rhamnose Links Moonlighting Proteins to Membrane Phospholipid in Mycoplasmas.

    PubMed

    Daubenspeck, James M; Liu, Runhua; Dybvig, Kevin

    2016-01-01

    Many proteins that have a primary function as a cytoplasmic protein are known to have the ability to moonlight on the surface of nearly all organisms. An example is the glycolytic enzyme enolase, which can be found on the surface of many types of cells from bacteria to human. Surface enolase is not enzymatic because it is monomeric and oligomerization is required for glycolytic activity. It can bind various molecules and activate plasminogen. Enolase lacks a signal peptide and the mechanism by which it attaches to the surface is unknown. We found that treatment of whole cells of the murine pathogen Mycoplasma pulmonis with phospholipase D released enolase and other common moonlighting proteins. Glycostaining suggested that the released proteins were glycosylated. Cytoplasmic and membrane-bound enolase was isolated by immunoprecipitation. No post-translational modification was detected on cytoplasmic enolase, but membrane enolase was associated with lipid, phosphate and rhamnose. Treatment with phospholipase released the lipid and phosphate from enolase but not the rhamnose. The site of rhamnosylation was identified as a glutamine residue near the C-terminus of the protein. Rhamnose has been found in all species of mycoplasma examined but its function was previously unknown. Mycoplasmas are small bacteria with have no peptidoglycan, and rhamnose in these organisms is also not associated with polysaccharide. We suggest that rhamnose has a central role in anchoring proteins to the membrane by linkage to phospholipid, which may be a general mechanism for the membrane association of moonlighting proteins in mycoplasmas and perhaps other bacteria. PMID:27603308

  11. Rhamnose Links Moonlighting Proteins to Membrane Phospholipid in Mycoplasmas

    PubMed Central

    Daubenspeck, James M.; Liu, Runhua; Dybvig, Kevin

    2016-01-01

    Many proteins that have a primary function as a cytoplasmic protein are known to have the ability to moonlight on the surface of nearly all organisms. An example is the glycolytic enzyme enolase, which can be found on the surface of many types of cells from bacteria to human. Surface enolase is not enzymatic because it is monomeric and oligomerization is required for glycolytic activity. It can bind various molecules and activate plasminogen. Enolase lacks a signal peptide and the mechanism by which it attaches to the surface is unknown. We found that treatment of whole cells of the murine pathogen Mycoplasma pulmonis with phospholipase D released enolase and other common moonlighting proteins. Glycostaining suggested that the released proteins were glycosylated. Cytoplasmic and membrane-bound enolase was isolated by immunoprecipitation. No post-translational modification was detected on cytoplasmic enolase, but membrane enolase was associated with lipid, phosphate and rhamnose. Treatment with phospholipase released the lipid and phosphate from enolase but not the rhamnose. The site of rhamnosylation was identified as a glutamine residue near the C-terminus of the protein. Rhamnose has been found in all species of mycoplasma examined but its function was previously unknown. Mycoplasmas are small bacteria with have no peptidoglycan, and rhamnose in these organisms is also not associated with polysaccharide. We suggest that rhamnose has a central role in anchoring proteins to the membrane by linkage to phospholipid, which may be a general mechanism for the membrane association of moonlighting proteins in mycoplasmas and perhaps other bacteria. PMID:27603308

  12. A serological investigation of Mycoplasma pneumoniae infection on the Witwatersrand.

    PubMed

    Joosting, A C; Harwin, R M; Coppin, A; Battaglia, P; van der Hoef, P

    1976-12-18

    Sera from patients with respiratory disease were examined for antibody to Mycoplasma pneumoniae by complement fixation test. During the study period of about 6 years, a 3-year cycle of infection was observed, which coincided with some epidemics in the UK and USA, suggesting the possibility of an approximately simultaneous world-wide spread. The epidemics lasted about 18 months each, during which the incidence of infection was over 10 times that of the interepidemic periods.

  13. Mycoplasma bovis mastitis and arthritis in a dairy heifer.

    PubMed

    2015-12-19

    Mycoplasma bovis causing mastitis and arthritis in a dairy heifer. Nutritional myopathy in a three-month-old suckler calf. Acute fasciolosis in ewes in Ayrshire. Cardiomyopathy of unknown aetiology causing death of a three-year-old Suffolk ram. Spinal aspergillosis in a seven-week-old pheasant poult These are among matters discussed in the disease surveillance report for August from SAC Consulting: Veterinary Services (SAC C VS).

  14. Mycoplasma bovis mastitis and arthritis in a dairy heifer.

    PubMed

    2015-12-19

    Mycoplasma bovis causing mastitis and arthritis in a dairy heifer. Nutritional myopathy in a three-month-old suckler calf. Acute fasciolosis in ewes in Ayrshire. Cardiomyopathy of unknown aetiology causing death of a three-year-old Suffolk ram. Spinal aspergillosis in a seven-week-old pheasant poult These are among matters discussed in the disease surveillance report for August from SAC Consulting: Veterinary Services (SAC C VS). PMID:26679914

  15. A fluorescent antibody technique for identification of Mycoplasma hyopneumoniae colonies.

    PubMed

    Schuller, W; Lehmkuhl, H D; Switzer, W P

    1976-04-01

    Fluorescent antibody staining of Mycoplasma hyopneumoniae colonies is difficult because the colonies wash from the agar. Airdrying of the colonies grown on solid medium-overlayed glass microscope slides fixed the colonies in place, so that fluorescent antibody stain could be readily accomplished. Apparent loss or alteration of antigenicity did not result from the air-drying process. The technique is useful for the identification of M hyopneumoniae isolates.

  16. The use of pulsed-field gel electrophoresis to investigate the epidemiology of Mycoplasma bovis in French calf feedlots.

    PubMed

    Arcangioli, Marie-Anne; Aslan, Hamidé; Tardy, Florence; Poumarat, François; Le Grand, Dominique

    2012-04-01

    Mycoplasma bovis is a major cause of respiratory outbreaks in cattle feedlots. In this study pulsed-field gel electrophoresis (PFGE) was used to trace field strains and provide information on M. bovis patterns of spread in calf feedlots. The suitability of KpnI, MluI and SmaI restriction enzymes was assessed on different sets of strains. The discriminative power of the first two enzymes was first assessed using 28 epidemiologically unrelated strains; stability was 100% on multiple isolates from in vivo experimental infection. Thirty-nine field isolates from six feedlots were then evaluated. In contrast to the unique fingerprints displayed by the unrelated strains, the isolates from the feedlots showed identical patterns at the time of the outbreak of respiratory disease and 4 weeks later. The PFGE typing results suggest that M. bovis strains follow a clonal epidemic spread pattern at the herd level and that the same strain persists in calves of the herd after the clinical signs have disappeared.

  17. Differential serologic response to Mycoplasma ovipneumoniae and Mycoplasma arginini in lambs affected with chronic respiratory disease.

    PubMed

    Niang, M; Rosenbusch, R F; Lopez-Virella, J; Kaeberle, M L

    1999-01-01

    An enzyme-linked immunoabsorbent assay (ELISA) was used to evaluate the levels of antibodies to Mycoplasma ovipneumoniae and M. arginini in lambs with chronic respiratory disease. Sera were obtained from lambs in several flocks at various stages of the clinical disease and tested with sodium dodecyl sulfate (SDS)-treated M. ovipneumoniae and M. arginini whole cells and a crude capsular extract of M. ovipneumoniae as the antigens. There were low levels of antibody to M. ovipneumoniae in flocks sampled at the early stages of infection, whereas increased levels of antibody were present in lambs from flocks that had apparently recovered from the clinical disease. Slowly rising titers of circulating antibodies to M. ovipneumoniae were confirmed by sequential bleeding of lambs during the course of the clinical disease. However, antibody levels of M. arginini were more likely to increase earlier in the disease process. There was significant cross-reactivity between the 2 SDS-treated antigens in both the ELISA test and western immunoblotting. In contrast, the crude capsular extract was specific for detecting antibodies to M. ovipneumoniae.

  18. Urease Color Test Medium U-9 for the Detection and Identification of “T” Mycoplasmas in Clinical Material

    PubMed Central

    Shepard, Maurice C.; Lunceford, Carl D.

    1970-01-01

    A urease color test fluid medium (U-9) for the detection and identification of T (T-strain) mycoplasmas in clinical material is described which is sensitive and specific for this group of mycoplasmas. The medium was prepared from commercially available components and contained 95% half-strength, tryptic digest broth (pH 5.5), 4% unheated horse serum, 0.05% highest-purity urea, 0.001% sodium phenolsulfonphthalein, and 1,000 units of potassium penicillin G per ml. The final reaction of medium U-9 was pH 6.0. The overall agreement (positive and negative) between urease reactions in U-9 urease color test medium and culture findings in a standard agar primary culture system among 686 clinical specimens was 98.1%. The disagreement consisted of 13 false-positive urease reactions which were recognized visually as false-positive reactions due to other microorganisms. For specimens from the female genitourinary tract, the inclusion of 2.5 μg of amphotericin B (Fungizone) per ml of medium U-9 is recommended for the suppression of growth of Candida species and filamentous fungi. PMID:4925243

  19. Comparison of antigens of pneumonia-associated mycoplasma species by gel diffusion.

    PubMed

    Ball, H J; Todd, D

    1978-09-01

    Comparison of fluorocarbon-extracted antigens of six mycoplasma species by double immunodiffusion and counterimmunodiffusion techniques revealed a close reciprocal relationship among Mycoplasma dispar, M. ovipneumoniae, and M. hyopneumoniae. A lesser degree of cross-reaction was also demonstrated between these three species and M. hyorhinis and M. bovoculi. The interrelationships were more clearly demonstrated by double immunodiffusion than by counterimmunodiffusion.

  20. Comparison of antigens of pneumonia-associated mycoplasma species by gel diffusion.

    PubMed Central

    Ball, H J; Todd, D

    1978-01-01

    Comparison of fluorocarbon-extracted antigens of six mycoplasma species by double immunodiffusion and counterimmunodiffusion techniques revealed a close reciprocal relationship among Mycoplasma dispar, M. ovipneumoniae, and M. hyopneumoniae. A lesser degree of cross-reaction was also demonstrated between these three species and M. hyorhinis and M. bovoculi. The interrelationships were more clearly demonstrated by double immunodiffusion than by counterimmunodiffusion. Images PMID:101469

  1. Experimental infection of BHK21 and Vero cell lines with different Mycoplasma spp

    PubMed Central

    Netto, Cristiane; Soccol, Vanete Thomaz; Sepulveda, Lya Madureira; Timenetsky, Jorge

    2014-01-01

    Mycoplasma spp, belongs to the class Mollicutes and is capable to produce alterations in cellular cultures causing damages to the biotechnological industry. Bioproducts generally require two essential inputs, bovine serum and cells. The study herein aims to evaluate the mycoplasma concentrations that affect the growing of BHK21 and Vero cells. The species used were: Mycoplasma orale, M. salivarium, M. arginini and M. hyorhinis, cultivated in a SP4 media. Two contamination tests were performed with BHK21 and Vero cells and one of them applied different concentrations of mycoplasma. In the first one, mycoplasma was applied at the day zero and, in the second one, the contamination was performed after the monolayer establishment. The both cellular cultures presented cytopathic effects with mycoplasma contamination, but the Vero cells suffered more damages than the BHK21 ones. It was also observed that the severity of the cytopathic effect depended on the mycoplasma specie, on the concentration and on the time of contact with the cellular culture, which evidences the importance of controlling the presence of mycoplasma in biotechnological industries. PMID:25763061

  2. 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

  3. Cross reactivity among the swine mycoplasmas as identified by protein microarray.

    PubMed

    Petersen, Andrew C; Oneal, David C; Seibel, Janice R; Poel, Kylie; Daum, Courtney L; Djordjevic, Steven P; Minion, F Chris

    2016-08-30

    Mycoplasmas are cell wall-less bacteria that infect a variety of animals in a species-specific manner. In swine, Mycoplasma hyopneumoniae is the most virulent and presents the most disease and economic problems to the swine industry. Serological assays are commonly used to assess colonization and disease, but antigenic cross-reactivity between M. hyopneumoniae and other mycoplasma species, most notably Mycoplasma hyorhinis, Mycoplasma hyosynoviae and Mycoplasma flocculare, is a concern. The extent of cross-reactivity has not been thoroughly investigated. These studies were designed to identify M. hyopneumoniae proteins that are recognized by rabbit hyperimmune sera raised against the other swine mycoplasmas. Our results indicate extensive cross-reactivity between M. flocculare and M. hyopneumoniae, which explains previous reports seen with ELISA assays. Only three of the thirty-nine M. hyopneumoniae proteins tested showed no cross reactivity with the other three swine mycoplasmas, mhp182 (42kDa C-terminal fragment), mhp638 and mhp684 (C-terminal fragment). Two proteins, mhp384 and mhp511, were cross-reactive with hyperimmune sera generated against three of the four species. None of the anti-M. hyorhinis hyperimmune sera reacted to any of the M. hyopneumoniae proteins. These results suggest that inapparent M. flocculare infections could produce positive responses in M. hyopneumoniae serological assays due to cross-reactivity, and that M. hyosynoviae infections are less likely to do so and M. hyorhinis infections unlikely to affect assay results. PMID:27527784

  4. World Health Organization International Standard To Harmonize Assays for Detection of Mycoplasma DNA.

    PubMed

    Nübling, C Micha; Baylis, Sally A; Hanschmann, Kay-Martin; Montag-Lessing, Thomas; Chudy, Michael; Kreß, Julia; Ulrych, Ursula; Czurda, Stefan; Rosengarten, Renate

    2015-09-01

    Nucleic acid amplification technique (NAT)-based assays (referred to here as NAT assays) are increasingly used as an alternative to culture-based approaches for the detection of mycoplasma contamination of cell cultures. Assay features, like the limit of detection or quantification, vary widely between different mycoplasma NAT assays. Biological reference materials may be useful for harmonization of mycoplasma NAT assays. An international feasibility study included lyophilized preparations of four distantly related mycoplasma species (Acholeplasma laidlawii, Mycoplasma fermentans, M. orale, M. pneumoniae) at different concentrations which were analyzed by 21 laboratories using 26 NAT assays with a qualitative, semiquantitative, or quantitative design. An M. fermentans preparation was shown to decrease the interassay variation when used as a common reference material. The preparation was remanufactured and characterized in a comparability study, and its potency (in NAT-detectable units) across different NATs was determined. The World Health Organization (WHO) Expert Committee on Biological Standardization (ECBS) established this preparation to be the "1st World Health Organization international standard for mycoplasma DNA for nucleic acid amplification technique-based assays designed for generic mycoplasma detection" (WHO Tech Rep Ser 987:42, 2014) with a potency of 200,000 IU/ml. This WHO international standard is now available as a reference preparation for characterization of NAT assays, e.g., for determination of analytic sensitivity, for calibration of quantitative assays in a common unitage, and for defining regulatory requirements in the field of mycoplasma testing.

  5. The effects of Mycoplasma hyopneumoniae on porcine circovirus type 2 replication in vitro PK-15 cells.

    PubMed

    Wang, Haiyan; Feng, Zhixin; Wu, Yuzi; Wei, Yanna; Gan, Yuan; Hua, Lizhong; Li, Bin; Wang, Xiaomin; Liu, Maojun; Xiong, Qiyan; Shao, Guoqing

    2016-04-01

    Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS). Mycoplasma hyopneumoniae (Mhp) is a very well-known co-factor that potentially enhances PCV2 replication and thus the development of PMWS. However, co-infection with Mhp and PCV2 in vivo under different conditions can produce divergent clinical signs and lesions. In this study, PCV2 replication could be enhanced by subsequent co-inoculation with Mhp (PCV2+Mhp) in a time and dose dependent method, but not by prior (Mhp+PCV2) or simultaneous (Mhp/PCV2) co-inoculation. Furthermore, different magnitudes of PCV2-infected cells, varying from 150% ± 14% to 351% ± 28%, were detected when co-infected with different Mhp strains. The relative percentage of PCV2-infected cells greatly decreased from 351% ± 28 to 141% ± 18 when the Mhp strain was treated with UV light for 12 h. These results offer the evidences to better understand the complex clinical syndromes in Mhp/PCV2 co-infection cases, and the occurrence of PMWS. PMID:27033909

  6. The functions of the variable lipoprotein family of Mycoplasma hyorhinis in adherence to host cells.

    PubMed

    Xiong, Qiyan; Wang, Jia; Ji, Yan; Ni, Bo; Zhang, Bixiong; Ma, Qinghong; Wei, Yanna; Xiao, Shaobo; Feng, Zhixin; Liu, Maojun; Shao, Guoqing

    2016-04-15

    Mycoplasma hyorhinis (M. hyorhinis) is a swine pathogen that is associated with various human cancers and contamination in cell cultures. However, no studies on the adhesion molecules of this pathogen have yet been reported. The variable lipoprotein (Vlp) family is an important surface component of M. hyorhinis. Herein, we performed several experiments to identify the function of the Vlp family in adherence to host cells. Seven recombinant Vlp (rVlp) proteins were expressed in Escherichia coli and purified by affinity chromatography. The potential role of rVlp adherence to pig kidney (PK-15) and swine tracheal epithelial (STEC) cells was then studied by indirect immunofluorescence assay and microtiter plate adherence assay. Adhesion of M. hyorhinis to PK-15 and STEC cells was specifically inhibited by the addition of a cocktail of rVlp proteins. The rVlp protein mixture was shown to bind to both PK-15 and STEC cells. The binding increased in a dose-dependent manner and could be blocked by antisera against the rVlp proteins. Most of the rVlp proteins could bind individually to both PK-15 and STEC cells except for rVlpD and rVlpF, which bound only to STEC cells. Because Vlp members vary in size among different strains and generations, they may vary in their cytoadhesion capabilities in various strains. In summary, the present results indicate that the Vlp family functions as adhesins of M. hyorhinis. PMID:27016761

  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.

  8. Development of a Multilocus Sequence Typing Scheme for Molecular Typing of Mycoplasma pneumoniae.

    PubMed

    Brown, Rebecca J; Holden, Matthew T G; Spiller, O Brad; Chalker, Victoria J

    2015-10-01

    Mycoplasma pneumoniae is a major human respiratory pathogen causing both upper and lower respiratory disease in humans of all ages, and it can also result in other serious extrapulmonary sequelae. A multilocus sequence typing (MLST) scheme for M. pneumoniae was developed based on the sequences of eight housekeeping genes (ppa, pgm, gyrB, gmk, glyA, atpA, arcC, and adk) and applied to 55 M. pneumoniae clinical isolates and the two type strains M129 and FH. A total of 12 sequence types (STs) resulted for 57 M. pneumoniae isolates tested, with a discriminatory index of 0.21 STs per isolate. The MLST loci used in this scheme were shown to be stable in 10 strains following 10 sequential subculture passages. Phylogenetic analysis of concatenated sequences of the eight loci indicated two distinct genetic clusters that were directly linked to multilocus variable-number tandem repeat analysis (MLVA) type. Genetic MLST clustering was confirmed by genomic sequence analysis, indicating that the MLST scheme developed in this study is representative of the genome. Furthermore, this MLST scheme was shown to be more discriminatory than both MLVA and P1 typing for the M. pneumoniae isolates examined, providing a method for further and more detailed analysis of observed epidemic peaks of M. pneumoniae infection. This scheme is supported by a public Web-based database (http://pubmlst.org/mpneumoniae).

  9. Occurrence of mycoplasmas in free-ranging birds of prey in Germany.

    PubMed

    Lierz, M; Hagen, N; Hernadez-Divers, S J; Hafez, H M

    2008-10-01

    Mycoplasmas are well-known avian pathogens of poultry and some passerines. Although reported in birds of prey, their role as pathogens is still unclear. Healthy, free-ranging raptor nestlings sampled during a routine ringing (banding) program, and birds of prey from rehabilitation centers, tested positive for Mycoplasma spp. by culture and a genus-specific polymerase chain reaction (PCR). Given the lack of clinical signs and disease, we suggest that mycoplasmas in raptors may be commensal rather than pathogenic. Using immunobinding assay and species-specific PCR tests, Mycoplasma buteonis, M. falconis, and M. gypis were identified; M. falconis was only detected in falcons. Additionally, some isolates could not be identified. This is the first report of Mycoplasma spp. isolations from Western Marsh Harriers (Circus aeroginosus), a Eurasian Hobby (Falco subbuteo), and a Barn Owl (Tyto alba). PMID:18957640

  10. [Molecular cloning and characterization of a N-acetylneuraminate lyase gene from Staphylococcus hominis].

    PubMed

    Zhou, Chuanhua; Chen, Xi; Feng, Jinhui; Xiao, Dongguang; Wuz, Qiaqing; Zhu, Dunming

    2013-04-01

    A N-acetylneuraminate lyase gene (shnal) from Staphylococcus hominis was cloned into pET-28a and expressed in Escherichia coli BL21 (DE3) host cells. The recombinant enzyme was purified and characterized. It is a homotetrameric enzyme with the optimum pH at 8.0 for the cleavage direction and the optimum pH and temperature were 7.5 and 45 degrees C for the synthetic direction. The activity of ShNAL is stable when incubated at 45 degrees C for 2 h but decreased rapidly over 50 degrees C. ShNAL showed high stability in a wide range pH from 5.0 to 10.0 with the residual activity being > 70% when the enzyme was incubated in different buffers at 4 degrees C for 24 h. Its K(m) towards N-acetylneuraminic acid, pyruvate and ManNAc were (4.0 +/- 0.2) mmol/L, (35.1 +/- 3.2) mmol/L and (131.7 +/- 12.1) mmol/L, respectively. The k(cat)/K(m) value of Neu5Ac, ManNAc, and Pyr for ShNAL were 1.9 L/(mmol x s), 0.08 L/(mmol x s) and 0.08 L/(mmol x s), respectively.

  11. Gastrospirillum Hominis associated chronic active gastritis: the first report from Italy.

    PubMed

    Ierardi, E; Monno, R; Mongelli, A; Allegretta, L; Milone, E; Rizzi, S; Panza, P; Coppolecchia, P; Francavilla, A

    1991-02-01

    Recently a spiral bacterium different from Helicobacter Pylori (HP) was observed in the human stomach and the name of Gastrospirillum Hominis (GH) was proposed for this organism. GH presence is reported to be not associated to HP but related to chronic active gastritis. We describe the case of a 31 year old male suffering from upper abdominal symptoms, who underwent oesophagogastroduodenoscopy, which revealed a picture of duodenal hyperemia. Gastric body showed a normal mucosa and absence of HP, while active chronic gastritis associated with HP was found in the antrum. In addition few spiral bacteria showing 4-5 spirals, larger than HP were observed within the gastric crypts and beneath the mucus layer in this site. This case represents the first report from our geographic area (Southern Italy) of the possibility of finding bacteria different from HP in the human stomach. The simultaneous HP presence does not allow us to relate the chronic active gastritis of the patient with the GH like bacteria. Our finding, however, suggests the possibility that HP and GH may be simultaneously present in the course of type B antral chronic inflammation. This association was not observed in previous investigations.

  12. Restriction enzyme digestion analysis of PCR-amplified DNA of Blastocystis hominis isolates.

    PubMed

    Init, I; Foead, A L; Fong, M Y; Yamazaki, H; Rohela, M; Yong, H S; Mak, J W

    2007-11-01

    Genomic DNA of Blastocystis isolates released into 0.1% Triton X-100 was suitable for amplification and yielded similar results as the genomic DNA extracted with standard kit. The specific B. hominis primers (BH1: GCT TAT CTG GTT GAT CCT GCC AGT and BH2: TGA TCC TTC CGC AGG TTC ACC TAC A) successfully produced the PCR product of about 1,770 bp with all the 7 Blastocystis isolates tested. The restriction fragment length polymorphism (RFLP) patterns yielded by 13 out of 25 restriction endonucleases showed that the 7 isolates could be grouped into 4 subgroups: subgroup-1 consisted of isolate C; subgroup-2 of isolates H4 and H7; subgroup-3 of isolates KP1, Y51 and M12; and subgroup-4 of isolate 27805. The differences between subgroups manifested as clear-cut RFLP patterns. A common band of 230 bp was revealed by Eco R1 in all the Blastocystis isolates tested. The band of about 180 bp was revealed by Alu I, differentiated symptomatic from asymptomatic isolates of this parasite, and might indicate the pathogenicity of this parasite. PMID:18613539

  13. Isolation of Mycoplasma gallopavonis from free-ranging wild turkeys in coastal North Carolina seropositive and culture-negative for Mycoplasma gallisepticum.

    PubMed

    Cobb, D T; Ley, D H; Doerr, P D

    1992-01-01

    Serum samples and choanal cleft swabs were collected from livetrapped and hunter killed wild turkeys (Meleagris gallopavo) from Martin and Bertie counties, North Carolina (USA). Sera were tested for antibodies to Mycoplasma gallisepticum, Mycoplasma synoviae and Mycoplasma meleagridis by hemagglutination inhibition (HI). Sera from 33% (five of 15) of livetrapped turkeys were positive for antibodies to M. gallisepticum by HI, and all were negative for antibodies to M. synoviae and M. meleagridis. Choanal cleft swabs from 22 livertrapped and five hunter killed wild turkeys cultured in Frey's broth medium resulted in 23 mycoplasma isolations. Using direct immunofluorescence, 74% (17/23) were M. gallopavonis, and 26% (six of 23) were unidentified; no isolate was identified as M. gallisepticum, M. synoviae or M. meleagridis.

  14. Two crystal structures of dihydrofolate reductase-thymidylate synthase from Cryptosporidium hominis reveal protein–ligand interactions including a structural basis for observed antifolate resistance

    SciTech Connect

    Anderson, Amy C.

    2005-03-01

    An analysis of the protein–ligand interactions in two crystal structures of DHFR-TS from C. hominis reveals a possible structural basis for observed antifolate resistance in C. hominis DHFR. A comparison with the structure of human DHFR reveals residue substitutions that may be exploited for the design of species-selective inhibitors. Cryptosporidium hominis is a protozoan parasite that causes acute gastrointestinal illness. There are no effective therapies for cryptosporidiosis, highlighting the need for new drug-lead discovery. An analysis of the protein–ligand interactions in two crystal structures of dihydrofolate reductase-thymidylate synthase (DHFR-TS) from C. hominis, determined at 2.8 and 2.87 Å resolution, reveals that the interactions of residues Ile29, Thr58 and Cys113 in the active site of C. hominis DHFR provide a possible structural basis for the observed antifolate resistance. A comparison with the structure of human DHFR reveals active-site differences that may be exploited for the design of species-selective inhibitors.

  15. The anti-mycoplasmal activity of aminosidine.

    PubMed

    Buogo, A; Zavaglio, V

    1975-12-01

    Some in vivo experiments of therapeutic activity in severe experimental infection (intracranial) in the mouse due to different strains of Mycoplasma proved the high efficacy of the antibiotic aminosidine when administered subcutaneously at the dose of 30 mg/kg and have also demonstrated that oral treatment is moderately effective against Mycoplasma hominis 2. Sensitivity test carried out in vitro on the same Mycoplasma strains confirmed the bactericidal activity of aminosidine. These experiments have therefore shown that the therapeutic spectrum of aminosidine, which includes Gram-positive, Gram-negative, acid-fast bacteria and some protozoa, may be enlarged to include the genus Mycoplasma. For this reason the antibiotic can find a useful application in the treatment of primary atypical neumonia. In addition, from a clinical research point of view, aminosidine may constitute a promising tool for the treatment of different illnesses in which Mycoplasma have been described as important co-factors of pathogenicity in man.

  16. The anti-mycoplasmal activity of aminosidine.

    PubMed

    Buogo, A; Zavaglio, V

    1975-12-01

    Some in vivo experiments of therapeutic activity in severe experimental infection (intracranial) in the mouse due to different strains of Mycoplasma proved the high efficacy of the antibiotic aminosidine when administered subcutaneously at the dose of 30 mg/kg and have also demonstrated that oral treatment is moderately effective against Mycoplasma hominis 2. Sensitivity test carried out in vitro on the same Mycoplasma strains confirmed the bactericidal activity of aminosidine. These experiments have therefore shown that the therapeutic spectrum of aminosidine, which includes Gram-positive, Gram-negative, acid-fast bacteria and some protozoa, may be enlarged to include the genus Mycoplasma. For this reason the antibiotic can find a useful application in the treatment of primary atypical neumonia. In addition, from a clinical research point of view, aminosidine may constitute a promising tool for the treatment of different illnesses in which Mycoplasma have been described as important co-factors of pathogenicity in man. PMID:1204831

  17. Mycoplasmas isolated from the respiratory tract of cattle and goats in Tanzania.

    PubMed

    Kusiluka, L J; Ojeniyi, B; Friis, N F; Kazwala, R R; Kokotovic, B

    2000-01-01

    A microbiological study of the mycoplasma flora in the respiratory tracts of cattle and goats in selected regions of Tanzania is described. In the examination of cattle, mycoplasmas were isolated from 60 (17.8%) of the 338 examined lung samples, 8 (47.1%) of the 17 lymph nodes, 4 (13.3%) of the 30 pleural fluid samples and 4 (3.9%) of the 103 nasal swabs examined. All the isolates were identified as Mycoplasma mycoides subsp. mycoides, Small Colony type except for one isolate from pleural fluid which was identified as Mycoplasma arginini. M. mycoides subsp. mycoides, Small Colony type was isolated from samples originating from Dodoma, Iringa, Mbeya, Morogoro and Shinyanga regions where outbreaks of contagious bovine pleuropneumonia had been reported. In the examination of goats, mycoplasmas were isolated from 54 (34.0%) of the 159 examined lung samples, 41 (18.1%) of the 226 nasal swabs and 4 (40.0%) of the 10 pleural fluid samples. The species demonstrated were Mycoplasma capricolum subsp. capripneumoniae, M. mycoides subsp. mycoides, Small Colony type Mycoplasma ovipneumoniae and M. Capricolum subsp. arginini. The isolation of M. capripneumoniae in the Coast and Morogoro regions confirmed the presence of contagious caprine pleuropneumonia in the regions.

  18. Isolation and identification of mycoplasmas from the nasal cavity of sheep.

    PubMed

    Brogden, K A; Rose, D; Cutlip, R C; Lehmkuhl, H D; Tully, J G

    1988-10-01

    Mycoplasmas isolated from the nasal cavity of sheep in a ram test station were examined to determine their identity and prevalence. Specimens were obtained for mycoplasmal culture in 1980, 1982, and 1983 from 558 sheep, and mycoplasmas were isolated from 630 specimens from 320 sheep (57.3%). The isolates were characterized and differentiated into groups on the basis of sensitivity to digitonin, fermentation of glucose, and hydrolysis of arginine. Isolates in some groups were further characterized by use of additional diagnostic media, and their identity was confirmed by agglutination or growth inhibition with antiserum prepared from reference mycoplasmas. Of the 320 sheep with mycoplasmas, 293 had Mycoplasma ovipneumoniae, 12 had M arginini, and 1 had M capricolum. Two sheep had Acholeplasma spp, and 3 sheep had unidentified Mycoplasma spp. The remaining 9 sheep had M ovipneumoniae in combination with Acholeplasma spp (n = 3), M arginini (n = 3), M capricolum (n = 2), and an unidentified Mycoplasma spp (n = 1). The biochemical reactions of the M ovipneumoniae from the 293 sheep were similar, but varied in the degree of growth and fermentation in the basal medium containing glucose. The high prevalence of M ovipneumoniae indicated that it may be commensal in the upper respiratory tract of healthy sheep.

  19. Expression of functions by normal sheep alveolar macrophages and their alteration by interaction with Mycoplasma ovipneumoniae.

    PubMed

    Niang, M; Rosenbusch, R F; Lopez-Virella, J; Kaeberle, M L

    1997-10-31

    Normal sheep alveolar macrophages collected by bronchial lavage were exposed to live or heat-killed Mycoplasma ovipneumoniae organisms, and their capability to ingest Staphylococcus aureus and to elicit antibody-dependent cellular cytotoxicity against sensitized chicken red blood cells was tested. Controls consisted of non-infected macrophages in M199 medium. In addition, the effect of M. ovipneumoniae on expression of surface molecules on these sheep alveolar macrophages was determined. The percentage of S. aureus ingested by nontreated sheep alveolar macrophages was significantly higher than that of infected macrophages. Live mycoplasmas were more effective in suppressing the ingestion of S. aureus by these macrophages than killed mycoplasmas. Both live and killed mycoplasmas suppressed the cytolytic effect of the sheep alveolar macrophages to a similar degree. About 78% and 45% of the normal sheep alveolar macrophages had IgG and complement receptors, respectively. Infection of these macrophages with M. ovipneumoniae decreased significantly the expression of IgG receptors but had no effects on complement receptors. There were substantial increases in the expression of both MHC class I and class II by the mycoplasma-induced macrophages as compared with unstimulated macrophages. Live mycoplasmas were more effective in inducing expression of both classes than killed mycoplasmas. The results, taken together, suggest that M. ovipneumoniae induced alterations in macrophage activities and this may be a contributing factor in the pathogenesis of respiratory disease induced by the organism.

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

    PubMed

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

    2013-03-01

    Three wild American black vultures (Coragyps atratus) were presented to rehabilitation centers with swelling of multiple joints, including elbows, stifles, hocks, and carpal joints, and of the gastrocnemius tendons. Cytological examination of the joint fluid exudate indicated heterophilic arthritis. Radiographic examination in 2 vultures demonstrated periarticular soft tissue swelling in both birds and irregular articular surfaces with subchondral bone erosion in both elbows in 1 bird. Prolonged antibiotic therapy administered in 2 birds did not improve the clinical signs. Necropsy and histological examination demonstrated a chronic lymphoplasmacytic arthritis involving multiple joints and gastrocnemius tenosynovitis. Articular lesions varied in severity and ranged from moderate synovitis and cartilage erosion and fibrillation to severe synovitis, diffuse cartilage ulceration, subchondral bone loss and/or sclerosis, pannus, synovial cysts, and epiphyseal osteomyelitis. No walled bacteria were observed or isolated from the joints. However, mycoplasmas polymerase chain reactions were positive in at least 1 affected joint from each bird. Mycoplasmas were isolated from joints of 1 vulture that did not receive antibiotic therapy. Sequencing of 16S rRNA gene amplicons from joint samples and the mycoplasma isolate identified Mycoplasma corogypsi in 2 vultures and was suggestive in the third vulture. Mycoplasma corogypsi identification was confirmed by sequencing the 16S-23S intergenic spacer region of mycoplasma isolates. This report provides further evidence that M. corogypsi is a likely cause of arthritis and tenosynovitis in American black vultures. Cases of arthritis and tenosynovitis in New World vultures should be investigated for presence of Mycoplasma spp, especially M. corogypsi.

  1. Genomic characterization of symbiotic mycoplasmas from the stomach of deep-sea isopod bathynomus sp.

    PubMed

    Wang, Yong; Huang, Jiao-Mei; Wang, Shao-Lu; Gao, Zhao-Ming; Zhang, Ai-Qun; Danchin, Antoine; He, Li-Sheng

    2016-09-01

    Deep-sea isopod scavengers such as Bathynomus sp. are able to live in nutrient-poor environments, which is likely attributable to the presence of symbiotic microbes in their stomach. In this study we recovered two draft genomes of mycoplasmas, Bg1 and Bg2, from the metagenomes of the stomach contents and stomach sac of a Bathynomus sp. sample from the South China Sea (depth of 898 m). Phylogenetic trees revealed a considerable genetic distance to other mycoplasma species for Bg1 and Bg2. Compared with terrestrial symbiotic mycoplasmas, the Bg1 and Bg2 genomes were enriched with genes encoding phosphoenolpyruvate-dependent phosphotransferase systems (PTSs) and sodium-driven symporters responsible for the uptake of sugars, amino acids and other carbohydrates. The genome of mycoplasma Bg1 contained sialic acid lyase and transporter genes, potentially enabling the bacteria to attach to the stomach sac and obtain organic carbons from various cell walls. Both of the mycoplasma genomes contained multiple copies of genes related to proteolysis and oligosaccharide degradation, which may help the host survive in low-nutrient conditions. The discovery of the different types of mycoplasma bacteria in the stomach of this deep-sea isopod affords insights into symbiotic model of deep-sea animals and genomic plasticity of mycoplasma bacteria. PMID:27312602

  2. Rapid imaging of mycoplasma in solution using Atmospheric Scanning Electron Microscopy (ASEM).

    PubMed

    Sato, Chikara; Manaka, Sachie; Nakane, Daisuke; Nishiyama, Hidetoshi; Suga, Mitsuo; Nishizaka, Takayuki; Miyata, Makoto; Maruyama, Yuusuke

    2012-01-27

    Mycoplasma is a genus of bacterial pathogen that causes disease in vertebrates. In humans, the species Mycoplasma pneumoniae causes 15% or more of community-acquired pneumonia. Because this bacterium is tiny, corresponding in size to a large virus, diagnosis using optical microscopy is not easy. In current methods, chest X-rays are usually the first action, followed by serology, PCR amplification, and/or culture, but all of these are particularly difficult at an early stage of the disease. Using Mycoplasma mobile as a model species, we directly observed mycoplasma in buffer with the newly developed Atmospheric Scanning Electron Microscope (ASEM). This microscope features an open sample dish with a pressure-resistant thin film window in its base, through which the SEM beam scans samples in solution, from below. Because of its 2-3μm-deep scanning capability, it can observe the whole internal structure of mycoplasma cells stained with metal solutions. Characteristic protein localizations were visualized using immuno-labeling. Cells were observed at low concentrations, because suspended cells concentrate in the observable zone by attaching to sialic acid on the silicon nitride (SiN) film surface within minutes. These results suggest the applicability of the ASEM for the study of mycoplasmas as well as for early-stage mycoplasma infection diagnosis. PMID:22226908

  3. Genomic characterization of symbiotic mycoplasmas from the stomach of deep-sea isopod bathynomus sp.

    PubMed

    Wang, Yong; Huang, Jiao-Mei; Wang, Shao-Lu; Gao, Zhao-Ming; Zhang, Ai-Qun; Danchin, Antoine; He, Li-Sheng

    2016-09-01

    Deep-sea isopod scavengers such as Bathynomus sp. are able to live in nutrient-poor environments, which is likely attributable to the presence of symbiotic microbes in their stomach. In this study we recovered two draft genomes of mycoplasmas, Bg1 and Bg2, from the metagenomes of the stomach contents and stomach sac of a Bathynomus sp. sample from the South China Sea (depth of 898 m). Phylogenetic trees revealed a considerable genetic distance to other mycoplasma species for Bg1 and Bg2. Compared with terrestrial symbiotic mycoplasmas, the Bg1 and Bg2 genomes were enriched with genes encoding phosphoenolpyruvate-dependent phosphotransferase systems (PTSs) and sodium-driven symporters responsible for the uptake of sugars, amino acids and other carbohydrates. The genome of mycoplasma Bg1 contained sialic acid lyase and transporter genes, potentially enabling the bacteria to attach to the stomach sac and obtain organic carbons from various cell walls. Both of the mycoplasma genomes contained multiple copies of genes related to proteolysis and oligosaccharide degradation, which may help the host survive in low-nutrient conditions. The discovery of the different types of mycoplasma bacteria in the stomach of this deep-sea isopod affords insights into symbiotic model of deep-sea animals and genomic plasticity of mycoplasma bacteria.

  4. Cryptosporidium hominis Is a Newly Recognized Pathogen in the Arctic Region of Nunavik, Canada: Molecular Characterization of an Outbreak

    PubMed Central

    Dixon, Brent; Dion, Réjean; Levesque, Benoît; Cantin, Philippe; Cédilotte, Lyne; Ndao, Momar; Proulx, Jean-François; Yansouni, Cedric P.

    2016-01-01

    Background Cryptosporidium is a leading cause of childhood diarrhea in low-resource settings, and has been repeatedly associated with impaired physical and cognitive development. In May 2013, an outbreak of diarrhea caused by Cryptosporidium hominis was identified in the Arctic region of Nunavik, Quebec. Human cryptosporidiosis transmission was previously unknown in this region, and very few previous studies have reported it elsewhere in the Arctic. We report clinical, molecular, and epidemiologic details of a multi-village Cryptosporidium outbreak in the Canadian Arctic. Methodology/Principal Findings We investigated the occurrence of cryptosporidiosis using a descriptive study of cases with onset between April 2013 and April 2014. Cases were defined as Nunavik inhabitants of any age presenting with diarrhea of any duration, in whom Cryptosporidium oocysts were detected by stool microscopy in a specialised reference laboratory. Cryptosporidium was identified in stool from 51 of 283 individuals. The overall annual incidence rate (IR) was 420 / 100,000 inhabitants. The IR was highest among children aged less than 5 years (1290 /100,000 persons). Genetic subtyping for stool specimens from 14/51 cases was determined by DNA sequence analysis of the 60 kDa glycoprotein (gp60) gene. Sequences aligned with C. hominis subtype Id in all cases. No common food or water source of infection was identified. Conclusions/Significance In this first observed outbreak of human cryptosporidiosis in this Arctic region, the high IR seen is cause for concern about the possible long-term effects on growth and development of children in Inuit communities, who face myriad other challenges such as overcrowding and food-insecurity. The temporal and geographic distribution of cases, as well as the identification of C. hominis subtype Id, suggest anthroponotic rather than zoonotic transmission. Barriers to timely diagnosis delayed the recognition of human cryptosporidiosis in this remote

  5. Mycoplasmas in wild turkeys living in association with domestic fowl.

    PubMed

    Hoffman, R W; Luttrell, M P; Davidson, W R; Ley, D H

    1997-07-01

    One hundred and nineteen Merriam's wild turkeys (Meleagris gallopavo merriami) and 31 domestic chickens coexisting on a ranch in west-central Colorado (USA) were surveyed for mycoplasmosis by serologic and cultural methods. Although no clinical signs were apparent in any wild turkeys tested, 51 (43%) had positive rapid plate agglutination (RPA) reactions for M. gallisepticum (MG) and/or M. synoviae (MS); 37% of 56 adults and 48% of 63 subadults were classified as positive reactors to MG and/or MS. No turkeys tested in 1992 (n = 61) and 17 (29%) of 58 turkeys tested in 1993 were RPA-positive for M. meleagridis (MM). Hemagglutination inhibition (HI) test results were negative for MG, MS and MM as were most enzyme-linked immunosorbent assay (ELISA) test reactions (MG = 99%, MS = 93%, MM = 87%). Immunoblotting showed mild to moderate reactivity to MG proteins in 49% of 41 samples tested. Most chickens were strongly positive for MS by RPA (81%), HI (58%) and ELISA (87%); 48% also were positive for MG by RPA but all were MG-negative by HI and ELISA. No pathogenic mycoplasmas were isolated from either group of birds. Mycoplasma gallopavonis was commonly identified from the wild turkeys, and M. gallinaceum was isolated from both the chickens and wild turkeys. In a transmission study conducted in 1994, disease-free domestic turkeys failed to seroconvert when co-housed with wild turkeys from this population that were RPA-positive for MG. Collectively, the results of this study were inconclusive regarding the status of pathogenic mycoplasmas within this wild turkey population.

  6. Monoclonal antibodies that inhibit mitogenic activity of Mycoplasma pulmonis.

    PubMed Central

    Lapidot, Z; Siman-Tov, R; Naot, Y

    1995-01-01

    Previous studies have suggested a correlation between mitogenic, polyclonal activation of host lymphocytes and the respiratory tract inflammatory diseases induced by Mycoplasma pulmonis. This study describes the generation of monoclonal antibodies (MAbs) to M. pulmonis membrane antigens with different capacities to inhibit stimulation of cultured rat lymphocytes by mycoplasmal membranes and with variable effects on M. pulmonis growth. We show that the inhibitory effects exerted on mitogenesis by purified MAbs are inversely related to the effects of MAbs on M. pulmonis growth. Immunoblotting of electrophoretically separated membrane proteins, with both growth- and mitogenesis-inhibiting antibodies, revealed significant changes in the reactions obtained with both types of MAb following short exposure of membranes to heat. Growth-inhibiting MAbs strongly react with heat-labile antigenic complexes with molecular weights of 65,000 to 75,000. Inhibition of mitogenesis is mainly associated with recognition of membrane complexes of 84 to 113 kDa that exhibit disperse smears and variable heat sensitivities. Following brief heating of membranes, more distinct bands of 103, 90, and 84 kDa are obtained with MAbs that inhibit mitogenesis. Experiments with other mitogenic mycoplasma species and MAb 3.3.10.2, a potent inhibitor of mitogenesis reveal that whereas the antigenic epitope recognized by this antibody is present on unheated membranes from different mycoplasmas, with heated membranes the MAb yields reactions only with M. pulmonis and M. arthritidis. Our studies suggest that M. pulmonis mitogens are unique membrane complexes of variable molecular weights, highly susceptible to heat and less sensitive to reducing agents. PMID:7806349

  7. Development and Evaluation of a Novel Single-Nucleotide-Polymorphism Real-Time PCR Assay for Rapid Detection of Fluoroquinolone-Resistant Mycoplasma bovis▿ †

    PubMed Central

    Ben Shabat, M.; Mikula, I.; Gerchman, I.; Lysnyansky, I.

    2010-01-01

    Monitoring of the susceptibility of Mycoplasma bovis field isolates to antibiotics is important for the appropriate choice of treatment. However, in vitro susceptibility testing of mycoplasmas is technically demanding and time-consuming, especially for clinical isolates, and is rarely performed in mycoplasma diagnostic laboratories. Thus, the development of methods allowing rapid real-time detection of resistant strains of M. bovis in clinical samples is a high priority for successful treatment. In this study, a novel TaqMan single-nucleotide-polymorphism (SNP) real-time PCR assay, which enables the rapid identification of M. bovis strains with different susceptibilities to fluoroquinolones, was developed and evaluated. The TaqMan SNP real-time PCR assay is based on the amplification of a 97-bp fragment of the parC quinolone resistance-determining region (QRDR) and allows the specific detection of four possible genotypes: GAC or GAT (susceptible to fluoroquinolones) and AAC or AAT (resistant to fluoroquinolones). Four TaqMan minor groove binder (MGB) probes identifying 1-base mismatches were designed and applied in a dual-probe assay with two reaction tubes. The TaqMan SNP real-time PCRs developed are highly specific for M. bovis, with a detection limit of 5 fg/μl (about 5 M. bovis genomes). In addition, all four SNP real-time PCR tests have almost the same efficiency (97.7% [GAC], 94% [AAC], 99.99% [GAT], and 98% [AAT]). Taken together, the data suggest that this SNP real-time PCR assay has potential as a routine diagnostic test for the detection of decreased susceptibility of M. bovis to fluoroquinolones. PMID:20534803

  8. MIB–MIP is a mycoplasma system that captures and cleaves immunoglobulin G

    PubMed Central

    Arfi, Yonathan; Minder, Laetitia; Di Primo, Carmelo; Le Roy, Aline; Ebel, Christine; Coquet, Laurent; Claverol, Stephane; Vashee, Sanjay; Jores, Joerg; Blanchard, Alain; Sirand-Pugnet, Pascal

    2016-01-01

    Mycoplasmas are “minimal” bacteria able to infect humans, wildlife, and a large number of economically important livestock species. Mycoplasma infections include a spectrum of clinical manifestations ranging from simple fever to fulminant inflammatory diseases with high mortality rates. These infections are mostly chronic, suggesting that mycoplasmas have developed means to evade the host immune response. Here we present and functionally characterize a two-protein system from Mycoplasma mycoides subspecies capri that is involved in the capture and cleavage of IgG. The first component, Mycoplasma Ig binding protein (MIB), is an 83-kDa protein that is able to tightly bind to the Fv region of a wide range of IgG. The second component, Mycoplasma Ig protease (MIP), is a 97-kDa serine protease that is able to cleave off the VH domain of IgG. We demonstrate that MIB is necessary for the proteolytic activity of MIP. Cleavage of IgG requires a sequential interaction of the different partners of the system: first MIB captures the IgG, and then MIP is recruited to the MIB–IgG complex, enabling protease activity. MIB and MIP are encoded by two genes organized in tandem, with homologs found in the majority of pathogenic mycoplasmas and often in multiple copies. Phylogenetic studies suggest that genes encoding the MIB–MIP system are specific to mycoplasmas and have been disseminated by horizontal gene transfer. These results highlight an original and complex system targeting the host immunoglobulins, playing a potentially key role in the immunity evasion by mycoplasmas. PMID:27114507

  9. MIB-MIP is a mycoplasma system that captures and cleaves immunoglobulin G.

    PubMed

    Arfi, Yonathan; Minder, Laetitia; Di Primo, Carmelo; Le Roy, Aline; Ebel, Christine; Coquet, Laurent; Claverol, Stephane; Vashee, Sanjay; Jores, Joerg; Blanchard, Alain; Sirand-Pugnet, Pascal

    2016-05-10

    Mycoplasmas are "minimal" bacteria able to infect humans, wildlife, and a large number of economically important livestock species. Mycoplasma infections include a spectrum of clinical manifestations ranging from simple fever to fulminant inflammatory diseases with high mortality rates. These infections are mostly chronic, suggesting that mycoplasmas have developed means to evade the host immune response. Here we present and functionally characterize a two-protein system from Mycoplasma mycoides subspecies capri that is involved in the capture and cleavage of IgG. The first component, Mycoplasma Ig binding protein (MIB), is an 83-kDa protein that is able to tightly bind to the Fv region of a wide range of IgG. The second component, Mycoplasma Ig protease (MIP), is a 97-kDa serine protease that is able to cleave off the VH domain of IgG. We demonstrate that MIB is necessary for the proteolytic activity of MIP. Cleavage of IgG requires a sequential interaction of the different partners of the system: first MIB captures the IgG, and then MIP is recruited to the MIB-IgG complex, enabling protease activity. MIB and MIP are encoded by two genes organized in tandem, with homologs found in the majority of pathogenic mycoplasmas and often in multiple copies. Phylogenetic studies suggest that genes encoding the MIB-MIP system are specific to mycoplasmas and have been disseminated by horizontal gene transfer. These results highlight an original and complex system targeting the host immunoglobulins, playing a potentially key role in the immunity evasion by mycoplasmas.

  10. Pilot study to evaluate the role of Mycoplasma species in cat bite abscesses.

    PubMed

    Torres-Henderson, Camille; Hesser, Jeff; Hyatt, Doreene R; Hawley, Jennifer; Brewer, Melissa; Lappin, Michael R

    2014-12-01

    Mycoplasma species are common inhabitants of the feline oral cavity, and so likely contaminate many cat bite abscesses. The objectives of this study were to determine whether Mycoplasma species are common contaminants of cat bite abscesses and whether they are are associated with β-lactam-resistant clinical disease. Twenty-six privately owned cats with clinical evidence of an abscess suspected to be from a cat bite were included in the study. Samples from each cat were evaluated by aerobic and anaerobic culture, as well as Mycoplasma species culture and polymerase chain reaction (PCR). All cats were initially treated with appropriate wound management and were administered an antibiotic of the β-lactam class (amoxicillin, amoxicillin clavulanate or cefovecin sodium). Mycoplasma species DNA was amplified by PCR from 4/26 samples (15.4%); one of these cases was concurrently culture positive. Adequate DNA for sequencing was present for 2/4 positive PCR samples; one was most homologous with Mycoplasma felis, and the other was most homologous with Mycoplasma equigenitalium and Mycoplasma elephantis. Of the 26 cats, 25 responded to the initial treatment by day 7. The cat that failed initial treatment was positive for M equigenitalium or M elephantis DNA on days 0 and 12, and ultimately responded to administration of enrofloxacin and clindamycin. The results suggest that while Mycoplasma species can contaminate cat bite abscesses, routine wound management and β-lactam antibiotic therapy is adequate for treatment in most cases of abscess. However, as Mycoplasma species infections do not respond to β-lactam class antibiotic therapy, these organisms should be on the differential list for cats with abscesses that fail treatment with this antibiotic class. PMID:24643287

  11. Pilot study to evaluate the role of Mycoplasma species in cat bite abscesses.

    PubMed

    Torres-Henderson, Camille; Hesser, Jeff; Hyatt, Doreene R; Hawley, Jennifer; Brewer, Melissa; Lappin, Michael R

    2014-12-01

    Mycoplasma species are common inhabitants of the feline oral cavity, and so likely contaminate many cat bite abscesses. The objectives of this study were to determine whether Mycoplasma species are common contaminants of cat bite abscesses and whether they are are associated with β-lactam-resistant clinical disease. Twenty-six privately owned cats with clinical evidence of an abscess suspected to be from a cat bite were included in the study. Samples from each cat were evaluated by aerobic and anaerobic culture, as well as Mycoplasma species culture and polymerase chain reaction (PCR). All cats were initially treated with appropriate wound management and were administered an antibiotic of the β-lactam class (amoxicillin, amoxicillin clavulanate or cefovecin sodium). Mycoplasma species DNA was amplified by PCR from 4/26 samples (15.4%); one of these cases was concurrently culture positive. Adequate DNA for sequencing was present for 2/4 positive PCR samples; one was most homologous with Mycoplasma felis, and the other was most homologous with Mycoplasma equigenitalium and Mycoplasma elephantis. Of the 26 cats, 25 responded to the initial treatment by day 7. The cat that failed initial treatment was positive for M equigenitalium or M elephantis DNA on days 0 and 12, and ultimately responded to administration of enrofloxacin and clindamycin. The results suggest that while Mycoplasma species can contaminate cat bite abscesses, routine wound management and β-lactam antibiotic therapy is adequate for treatment in most cases of abscess. However, as Mycoplasma species infections do not respond to β-lactam class antibiotic therapy, these organisms should be on the differential list for cats with abscesses that fail treatment with this antibiotic class.

  12. MIB-MIP is a mycoplasma system that captures and cleaves immunoglobulin G.

    PubMed

    Arfi, Yonathan; Minder, Laetitia; Di Primo, Carmelo; Le Roy, Aline; Ebel, Christine; Coquet, Laurent; Claverol, Stephane; Vashee, Sanjay; Jores, Joerg; Blanchard, Alain; Sirand-Pugnet, Pascal

    2016-05-10

    Mycoplasmas are "minimal" bacteria able to infect humans, wildlife, and a large number of economically important livestock species. Mycoplasma infections include a spectrum of clinical manifestations ranging from simple fever to fulminant inflammatory diseases with high mortality rates. These infections are mostly chronic, suggesting that mycoplasmas have developed means to evade the host immune response. Here we present and functionally characterize a two-protein system from Mycoplasma mycoides subspecies capri that is involved in the capture and cleavage of IgG. The first component, Mycoplasma Ig binding protein (MIB), is an 83-kDa protein that is able to tightly bind to the Fv region of a wide range of IgG. The second component, Mycoplasma Ig protease (MIP), is a 97-kDa serine protease that is able to cleave off the VH domain of IgG. We demonstrate that MIB is necessary for the proteolytic activity of MIP. Cleavage of IgG requires a sequential interaction of the different partners of the system: first MIB captures the IgG, and then MIP is recruited to the MIB-IgG complex, enabling protease activity. MIB and MIP are encoded by two genes organized in tandem, with homologs found in the majority of pathogenic mycoplasmas and often in multiple copies. Phylogenetic studies suggest that genes encoding the MIB-MIP system are specific to mycoplasmas and have been disseminated by horizontal gene transfer. These results highlight an original and complex system targeting the host immunoglobulins, playing a potentially key role in the immunity evasion by mycoplasmas. PMID:27114507

  13. Performance of PCR-based and Bioluminescent assays for mycoplasma detection.

    PubMed

    Falagan-Lotsch, Priscila; Lopes, Talíria Silva; Ferreira, Nívea; Balthazar, Nathália; Monteiro, Antônio M; Borojevic, Radovan; Granjeiro, José Mauro

    2015-11-01

    Contaminated eukaryotic cell cultures are frequently responsible for unreliable results. Regulatory entities request that cell cultures must be mycoplasma-free. Mycoplasma contamination remains a significant problem for cell cultures and may have an impact on biological analysis since they affect many cell parameters. The gold standard microbiological assay for mycoplasma detection involves laborious and time-consuming protocols. PCR-based and Bioluminescent assays have been considered for routine cell culture screening in research laboratories since they are fast, easy and sensitive. Thus, the aim of this work is to compare the performance of two popular commercial assays, PCR-based and Bioluminescent assays, by assessing the level of mycoplasma contamination in cell cultures from Rio de Janeiro Cell Bank (RJCB) and also from customers' laboratories. The results obtained by both performed assays were confirmed by scanning electron microscopy. In addition, we evaluated the limit of detection of the PCR kit under our laboratory conditions and the storage effects on mycoplasma detection in frozen cell culture supernatants. The performance of both assays for mycoplasma detection was not significantly different and they showed very good agreement. The Bioluminescent assay for mycoplasma detection was slightly more dependable than PCR-based due to the lack of inconclusive results produced by the first technique, especially considering the ability to detect mycoplasma contamination in frozen cell culture supernatants. However, cell lines should be precultured for four days or more without antibiotics to obtain safe results. On the other hand, a false negative result was obtained by using this biochemical approach. The implementation of fast and reliable mycoplasma testing methods is an important technical and regulatory issue and PCR-based and Bioluminescent assays may be good candidates. However, validation studies are needed. PMID:26296900

  14. Rapid imaging of mycoplasma in solution using Atmospheric Scanning Electron Microscopy (ASEM)

    SciTech Connect

    Sato, Chikara; Manaka, Sachie; Nakane, Daisuke; Nishiyama, Hidetoshi; Suga, Mitsuo; Nishizaka, Takayuki; Miyata, Makoto; Maruyama, Yuusuke

    2012-01-27

    Highlights: Black-Right-Pointing-Pointer Mycoplasma mobile was observed in buffer with the Atmospheric Scanning Electron Microscope. Black-Right-Pointing-Pointer Characteristic protein localizations were visualized using immuno-labeling. Black-Right-Pointing-Pointer M. mobile attached to sialic acid on the SiN film surface within minutes. Black-Right-Pointing-Pointer Cells were observed at low concentrations. Black-Right-Pointing-Pointer ASEM should promote study and early-stage diagnosis of mycoplasma. -- Abstract: Mycoplasma is a genus of bacterial pathogen that causes disease in vertebrates. In humans, the species Mycoplasma pneumoniae causes 15% or more of community-acquired pneumonia. Because this bacterium is tiny, corresponding in size to a large virus, diagnosis using optical microscopy is not easy. In current methods, chest X-rays are usually the first action, followed by serology, PCR amplification, and/or culture, but all of these are particularly difficult at an early stage of the disease. Using Mycoplasma mobile as a model species, we directly observed mycoplasma in buffer with the newly developed Atmospheric Scanning Electron Microscope (ASEM). This microscope features an open sample dish with a pressure-resistant thin film window in its base, through which the SEM beam scans samples in solution, from below. Because of its 2-3 {mu}m-deep scanning capability, it can observe the whole internal structure of mycoplasma cells stained with metal solutions. Characteristic protein localizations were visualized using immuno-labeling. Cells were observed at low concentrations, because suspended cells concentrate in the observable zone by attaching to sialic acid on the silicon nitride (SiN) film surface within minutes. These results suggest the applicability of the ASEM for the study of mycoplasmas as well as for early-stage mycoplasma infection diagnosis.

  15. First report of furuncular myiasis caused by the larva of botfly, Dermatobia hominis, in a Taiwanese traveler

    PubMed Central

    Hu, Je-Ming; Wang, Chih-Chien; Chao, Li-Lian; Lee, Chung-Shinn; Shih, Chien-Ming

    2013-01-01

    A case of furuncular myiasis was reported for the first time in a 29-year-old young Taiwanese traveler returning from an ecotourism in Peru. Furuncle-like lesions were observed on the top of his head and he complained of crawling sensations within his scalp. The invasive larva of botfly, Dermatobia hominis, was extruded from the furuncular lesion of the patient. Awareness of cutaneous myiasis for clinicians should be considered for a patient who has a furuncular lesion and has recently returned from a botfly-endemic area. PMID:23620844

  16. First report of furuncular myiasis caused by the larva of botfly, Dermatobia hominis, in a Taiwanese traveler.

    PubMed

    Hu, Je-Ming; Wang, Chih-Chien; Chao, Li-Lian; Lee, Chung-Shinn; Shih, Chien-Ming

    2013-03-01

    A case of furuncular myiasis was reported for the first time in a 29-year-old young Taiwanese traveler returning from an ecotourism in Peru. Furuncle-like lesions were observed on the top of his head and he complained of crawling sensations within his scalp. The invasive larva of botfly, Dermatobia hominis, was extruded from the furuncular lesion of the patient. Awareness of cutaneous myiasis for clinicians should be considered for a patient who has a furuncular lesion and has recently returned from a botfly-endemic area. PMID:23620844

  17. First report of furuncular myiasis caused by the larva of botfly, Dermatobia hominis, in a Taiwanese traveler.

    PubMed

    Hu, Je-Ming; Wang, Chih-Chien; Chao, Li-Lian; Lee, Chung-Shinn; Shih, Chien-Ming

    2013-03-01

    A case of furuncular myiasis was reported for the first time in a 29-year-old young Taiwanese traveler returning from an ecotourism in Peru. Furuncle-like lesions were observed on the top of his head and he complained of crawling sensations within his scalp. The invasive larva of botfly, Dermatobia hominis, was extruded from the furuncular lesion of the patient. Awareness of cutaneous myiasis for clinicians should be considered for a patient who has a furuncular lesion and has recently returned from a botfly-endemic area.

  18. Mycoplasma felis pleuritis in two show-jumper horses.

    PubMed

    Hoffman, A M; Baird, J D; Kloeze, H J; Rosendal, S; Bell, M

    1992-04-01

    Mycoplasma felis was identified as the cause of acute pleuritis in 2 show-jumping horses. The pleural exudate was proteinaceous, contained large numbers of neutrophils, and had a markedly increased lactate concentration. M. felis was isolated in pure culture from pleural fluid. Rising serum antibody titers to M. felis as well as a precipitous decline in titers to equine influenza virus were demonstrated in both horses. Pleural effusion in both horses and a pneumothorax detected in one of the horses resolved following a single drainage of pleural fluid and intravenous fluid, antibiotic, and analgesic therapy. PMID:1623728

  19. [Natural and experimental infections of lambs with Mycoplasma ovipneumoniae].

    PubMed

    Bocklisch, H; Pfützner, H; Zepezauer, V

    1989-01-01

    Mycoplasma (M.) ovipneumoniae was isolated pure or mixed with bacteria from 47 lungs of lambs of 14 in 22 tested flocks. M. ovipneumoniae was obtained as pure culture in cases of mild bronchopneumonia. Experimental intratracheal or intranasal infection caused several days of rising body temperature above 39.7 degrees C. Nasal discharge, coughing, and dyspnea did not occur. M. ovipneumoniae was successfully re-isolated from nasal swabs, beginning 2 d from infection. Lobular catarrhal bronchopneumonia was established by postmortem examinations, 10-14 d from infection, and M. ovipneumoniae was re-isolated from the lungs. Histological patterns of lungs were characterised by interstitial cell reactions.

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

    PubMed

    Cookson, K C; Shivaprasad, H L

    1994-01-01

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