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Sample records for equi em doente

  1. Rhodococcus equi foal pneumonia.

    PubMed

    Cohen, Noah D

    2014-12-01

    Pneumonia caused by Rhodococcus equi is an important cause of disease and death in foals. This article reviews current knowledge of the epidemiology, clinical signs, diagnosis, treatment, prevention, and control of R equi pneumonia in foals. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Update on Streptococcus equi subsp equi infections.

    PubMed

    Mallicote, Martha

    2015-04-01

    There are few diseases that ignite as much fervor among horse owners as strangles. Streptococcus equi subsp equi (strangles) infections frequently require the treating veterinarian to manage not only the clinical cases but also the biosecurity and provision of information to all involved parties. Although the disease is typically characterized by low mortality and high morbidity, restrictions of horse movement that result from appropriate quarantine procedures often frustrate the involved parties. The aims of this article are to provide clinically relevant information for diagnosis, treatment, and biosecurity management of strangles infection.

  3. Rhodococcus equi (Prescottella equi) vaccines; the future of vaccine development.

    PubMed

    Giles, C; Vanniasinkam, T; Ndi, S; Barton, M D

    2015-09-01

    For decades researchers have been targeting prevention of Rhodococcus equi (Rhodococcus hoagui/Prescottella equi) by vaccination and the horse breeding industry has supported the ongoing efforts by researchers to develop a safe and cost effective vaccine to prevent disease in foals. Traditional vaccines including live, killed and attenuated (physical and chemical) vaccines have proved to be ineffective and more modern molecular-based vaccines including the DNA plasmid, genetically attenuated and subunit vaccines have provided inadequate protection of foals. Newer, bacterial vector vaccines have recently shown promise for R. equi in the mouse model. This article describes the findings of key research in R. equi vaccine development and looks at alternative methods that may potentially be utilised.

  4. Streptococcus equi subspecies equi infection (strangles) in horses.

    PubMed

    Boyle, Ashley

    2011-03-01

    Streptococcus equi subspecies equi (strangles) is a highly contagious upper respiratory infection in horses. The infection is transmitted by inhalation or direct contact with mucopurulent discharge from an infective animal, resulting in fever, depression, and submandibular and retropharyngeal lymph node enlargement that can lead to respiratory distress. Complications include purpura hemorrhagica and metastatic abscessation. Control of outbreaks requires strict isolation protocols and hygiene measures. Detection of carriers is essential for preventing disease recurrence on a farm.

  5. Genetic Susceptibility to Rhodococcus equi.

    PubMed

    McQueen, C M; Dindot, S V; Foster, M J; Cohen, N D

    2015-01-01

    Rhodococcus equi pneumonia is a major cause of morbidity and mortality in neonatal foals. Much effort has been made to identify preventative measures and new treatments for R. equi with limited success. With a growing focus in the medical community on understanding the genetic basis of disease susceptibility, investigators have begun to evaluate the interaction of the genetics of the foal with R. equi. This review describes past efforts to understand the genetic basis underlying R. equi susceptibility and tolerance. It also highlights the genetic technology available to study horses and describes the use of this technology in investigating R. equi. This review provides readers with a foundational understanding of candidate gene approaches, single nucleotide polymorphism-based, and copy number variant-based genome-wide association studies, and next generation sequencing (both DNA and RNA). Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  6. Rhodococcus equi Infections in Dogs.

    PubMed

    Bryan, L K; Clark, S D; Díaz-Delgado, J; Lawhon, S D; Edwards, J F

    2017-01-01

    Five cases of Rhodococcus equi infection in dogs were identified from 2003 to 2014. Three of the dogs had severe, internal lesions attributable to R. equi that have not been previously described: endophthalmitis, endocarditis, and suppurative pleuropneumonia. Isolates from 4 of the dogs were analyzed by polymerase chain reaction for Rhodococcus virulence-associated plasmid (vap) genes. One isolate was vapA-positive, 2 lacked a virulence plasmid, and 1 carried the novel vapN-associated plasmid (pVAPN) recently characterized in bovine isolates. The pVAPN plasmid has not been described in isolates cultured from companion animals. Four of the dogs either were receiving immunosuppressive drugs or had endocrinopathies. R. equi has the potential to cause significant infections in dogs, and immunocompromised animals should be considered at risk for infection.

  7. Improved electroporation of Rhodococcus equi.

    PubMed

    Sekizaki, T; Tanoue, T; Osaki, M; Shimoji, Y; Tsubaki, S; Takai, S

    1998-02-01

    The condition of an electroporation method was re-evaluated for the introduction of foreign plasmid DNA into Rhodococcus equi. The method is based on an electroporation of the bacteria made competent by culturing in a broth containing glycine and by heat shock at 50 degrees C. Transformation of R. equi could be achieved with a chloramphenicol-resistant shuttle vector originating from Rhodococcus fascians at an efficiency of about 10(4) transformants/microgram DNA. The bacteria were also shown to become competent when they were incubated with a chemical transformation buffer prior to washing with an electroporation buffer.

  8. Streptococcus equi meningoencephalomyelitis in a foal.

    PubMed

    Finno, Carrie; Pusterla, Nicola; Aleman, Monica; Mohr, F Charles; Price, Teena; George, Jeanne; Holmberg, Tara

    2006-09-01

    A 4-month-old American Paint Horse colt was evaluated because of acute onset of ataxia, left-sided head tilt, and fever and a recently noticed heart murmur. Upper respiratory tract infection caused by Streptococcus equi subsp equi had been diagnosed at 3 months of age. Hematologic abnormalities included leukocytosis, mature neutrophilia, monocytosis, and mild anemia. Analysis of a CSF sample revealed high total protein concentration and total nucleated cell count; nucleated cells consisted mainly of degenerate neutrophils. Results of a real-time PCR assay were positive for S equi subsp equi, and a diagnosis of S equi subsp equi meningoencephalomyelitis was made. Treatment included administration of potassium penicillin and fluids, but the foal developed uroperitoneum and was subsequently euthanized. Postmortem examination revealed meningoencephalomyelitis, and S equi subsp equi was cultured from a brain aspirate. Additional findings included suppurative cystitis with rupture and neutrophilic myocarditis. Findings suggest that S equi subsp equi meningoencephalomyelitis should be considered in the differential diagnosis for foals with neurologic signs that have a history of strangles or exposure to affected horses.

  9. Treatment of Infections Caused by Rhodococcus equi.

    PubMed

    Giguère, Steeve

    2017-04-01

    Pneumonia caused by Rhodococcus equi remains an important cause of disease and death in foals. The combination of a macrolide (erythromycin, azithromycin, or clarithromycin) with rifampin remains the recommended therapy for foals with clinical signs of infection caused by R equi. Most foals with small, subclinical ultrasonographic pulmonary lesions associated with R equi recover without therapy, and administration of antimicrobial agents to these subclinically affected foals does not hasten lesion resolution relative to administration of a placebo. Resistance to macrolides and rifampin in isolates of R equi is increasing. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Molecular characterization of virulence genes of Streptococcus equi subsp. equi and Streptococcus equi subsp. zooepidemicus in equines

    PubMed Central

    Javed, R.; Taku, A. K.; Gangil, Rakhi; Sharma, R. K.

    2016-01-01

    Aim: The aim was to determine the occurrence of streptococci in equines in Jammu (R. S. Pura, Katra), characterization of Streptococci equi subsp. equi and Streptococcus equi subsp. zooepidemicus with respect to their virulence traits and to determine antibiotic sensitivity pattern of virulent Streptococcus isolates. Materials and Methods: A total of 96 samples were collected from both clinically affected animals (exhibiting signs of respiratory tract disease) and apparently healthy animals and were sent to laboratory. The organisms were isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and confirmed by cultural characteristics and biochemical tests. Molecular detection of Streptococcus was done directly from cultures using sodA and seM gene-based polymerase chain reaction (PCR). Antibiogram was performed against five antibiotics such as amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. Results: During this study, a total 40 streptococcal isolates were obtained out of which 2 isolates were of S. equi subsp. equi, 12 isolates were from S. equi subsp. zooepidemicus. In the PCR-based detection, we revealed amplicons of 235 bp and 679 bp for confirmation of sodA and seM gene, respectively. In antibiogram, two isolates of S. equi subsp. equi were found resistant to penicillin G, and all other isolates were found sensitive to amoxicillin and streptomycin. Conclusion: The majority of streptococcal infections was due to S. equi subsp. Zooepidemicus, and thus was recognized as a potential pathogen of diseases of equines besides S. equi subsp. equi. PMID:27651677

  11. Antimicrobial resistance in Rhodococcus equi.

    PubMed

    Cisek, Agata A; Rzewuska, Magdalena; Witkowski, Lucjan; Binek, Marian

    2014-01-01

    Rhodococcus equi is an important etiologic agent of respiratory- and non-respiratory tract infections, diseases of animals and humans. Therapy includes the use of various group of chemotherapeutic agents, however resistance acquirement is quite common. To date there is no preferred treatment protocol for infections caused by isolates resistant to macrolides and rifampicin. The resistance acquirement is a result of many molecular mechanisms, some of which include alterations in the cell envelope composition and structure, activity of the efflux pumps, enzymatic destruction or inactivation of antibiotics, and changes in the target site. This paper contains an overview of antimicrobial susceptibility of R. equi, and explains the possible molecular mechanisms responsible for antimicrobial resistance in this particular microorganism.

  12. Immunologically reactive proteins of Streptococcus equi.

    PubMed Central

    Timoney, J F; Trachman, J

    1985-01-01

    Immunologically reactive proteins in acid extracts and culture supernatants of Streptococcus equi were recognized through a combination of chromatographic and immunologic procedures. Both high- and low-molecular-weight components of each of these protein preparations were protective for mice and were, therefore, presumed to contain a variety of hydrolytic products or fragments of the M protein of S. equi. Convalescent horse sera that exhibited strong bactericidal activity for S. equi always reacted with polypeptides in the molecular weight range of 24,000 to 29,000, whereas preinfection sera did not. Rabbit antisera to affinity-purified S. equi protein also reacted with these polypeptides, as well as with a polypeptide of about 36,000 to 37,000 molecular weight. M protein in acid extract and culture supernatant did not cross-react in immunodiffusion, but rabbit antiserum to affinity-purified M protein from an acid extract of S. equi reacted strongly with culture supernatant proteins of approximate molecular weights of 67,000, 58,000, and 43,000. We suggest, therefore, that the M protein in culture supernatant is masked by other sequences that are removed by hot acid during preparation of acid extracts. Polypeptides common to acid extracts of S. equi and Streptococcus zooepidemicus were also identified. These polypeptides had molecular weights of about 55,000 and 31,000. Images PMID:3980091

  13. Quantitative aspects of fecal Rhodococcus (Corynebacterium) equi in foals.

    PubMed Central

    Takai, S; Ohkura, H; Watanabe, Y; Tsubaki, S

    1986-01-01

    Quantitative aspects of fecal Rhodococcus (Corynebacterium) equi in newborn foals for 12 weeks after birth were investigated on two horse breeding farms. R. equi was found in the feces of foals during week 1 of life. The greatest numbers of R. equi were present in the feces of foals during the first 8 weeks of their lives, which coincides with the age when foals are most liable to be exposed to R. equi. PMID:3700632

  14. Study of lysozyme resistance in Rhodococcus equi.

    PubMed

    Hébert, Laurent; Bidaud, Pauline; Goux, Didier; Benachour, Abdellah; Laugier, Claire; Petry, Sandrine

    2014-03-01

    Lysozyme is an important and widespread component of the innate immune response that constitutes the first line of defense against bacterial pathogens. The bactericidal effect of this enzyme relies on its capacity to hydrolyze the bacterial cell wall and also on a nonenzymatic mechanism involving its cationic antimicrobial peptide (CAMP) properties, which leads to membrane permeabilization. In this paper, we report our findings on the lysozyme resistance ability of Rhodococcus equi, a pulmonary pathogen of young foals and, more recently, of immunocompromised patients, whose pathogenic capacity is conferred by a large virulence plasmid. Our results show that (i) R. equi can be considered to be moderately resistant to lysozyme, (ii) the activity of lysozyme largely depends on its muramidase action rather than on its CAMP activity, and (iii) the virulence plasmid confers part of its lysozyme resistance capacity to R. equi. This study is the first one to demonstrate the influence of the virulence plasmid on the stress resistance capacity of R. equi and improves our understanding of the mechanisms enabling R. equi to resist the host defenses.

  15. Rhodococcus equi: an animal and human pathogen.

    PubMed Central

    Prescott, J F

    1991-01-01

    Recent isolations of Rhodococcus equi from cavitatory pulmonary disease in patients with AIDS have aroused interest among medical microbiologists in this unusual organism. Earlier isolations from humans had also been in immunosuppressed patients following hemolymphatic tumors or renal transplantation. This organism has been recognized for many years as a cause of a serious pyogranulomatous pneumonia of young foals and is occasionally isolated from granulomatous lesions in several other species, in some cases following immunosuppression. The last decade has seen many advances in understanding of the epidemiology, pathogenesis, diagnosis, treatment, and immunity to infection in foals. The particular susceptibility of the foal is not understood but can be explained in part by a combination of heavy challenge through the respiratory route coinciding with declining maternally derived antibody in the absence of fully competent foal cellular immune mechanisms. R. equi is largely a soil organism but is widespread in the feces of herbivores. Its growth in soil is considerably improved by simple nutrients it obtains from herbivore manure. About one-third of human patients who have developed R. equi infections had contact in some way with herbivores or their manure. Others may have acquired infection from contact with soil or wild bird manure. R. equi is an intracellular parasite, which explains the typical pyogranulomatous nature of R. equi infections, the predisposition to infection in human patients with defective cell-mediated immune mechanisms, and the efficacy of antimicrobial drugs that penetrate phagocytic cells. Images PMID:2004346

  16. Association of perinatal exposure to airborne Rhodococcus equi with risk of pneumonia caused by R equi in foals.

    PubMed

    Cohen, Noah D; Chaffin, M Keith; Kuskie, Kyle R; Syndergaard, Melissa K; Blodgett, Glenn P; Takai, Shinji

    2013-01-01

    To determine whether the concentrations of airborne virulent Rhodococcus equi in stalls housing foals during the first 2 weeks after birth are associated with subsequent development of R equi pneumonia in those foals. Air samples collected from foaling stalls and holding pens in which foals were housed during the first 2 weeks after birth. At a breeding farm in Texas, air samples (500 L each) were collected (January through May 2011) from stalls and pens in which 121 foals were housed on day 1 and on days 4, 7, and 14 after birth. For each sample, the concentration of airborne virulent R equi was determined with an immunoblot technique. The association between development of pneumonia and airborne R equi concentration was evaluated via random-effects Poisson regression analysis. Some air samples were not available for analysis. Of the 471 air samples collected from stalls that housed 121 foals, 90 (19%) contained virulent R equi. Twenty-four of 121 (20%) foals developed R equi pneumonia. Concentrations of virulent R equi in air samples from stalls housing foals that developed R equi pneumonia were significantly higher than those in samples from stalls housing foals that did not develop pneumonia. Accounting for disease effects, air sample concentrations of virulent R equi did not differ significantly by day after birth or by month of birth. Exposure of foals to airborne virulent R equi during the first 2 weeks after birth was significantly (and likely causally) associated with development of R equi pneumonia.

  17. Streptococcus equi subsp. zooepidemicus meningitis in Peru.

    PubMed

    Mori, Nicanor; Guevara, Jose M; Tilley, Drake H; Briceno, Jesus A; Zunt, Joseph R; Montano, Silvia M

    2013-02-01

    A 59-year-old man with a history of fever, unsteadiness, hemiparesis, motor aphasia and consciousness disturbance was hospitalized for Streptococcus equi subsp. zooepidemicus meningitis. He denied contact with farm animals, but had a practice of consuming unpasteurized goats' cheese from an uncertain source.

  18. Streptococcus equi subsp. zooepidemicus meningitis in Peru

    PubMed Central

    Guevara, Jose M.; Tilley, Drake H.; Briceno, Jesus A.; Zunt, Joseph R.; Montano, Silvia M.

    2013-01-01

    A 59-year-old man with a history of fever, unsteadiness, hemiparesis, motor aphasia and consciousness disturbance was hospitalized for Streptococcus equi subsp. zooepidemicus meningitis. He denied contact with farm animals, but had a practice of consuming unpasteurized goats’ cheese from an uncertain source. PMID:23105024

  19. Diversity of seM in Streptococcus equi subsp. equi isolated from strangles outbreaks.

    PubMed

    Libardoni, Felipe; Vielmo, Andréia; Farias, Luana; Matter, Letícia Beatriz; Pötter, Luciana; Spilki, Fernando Rosado; de Vargas, Agueda Castagna

    2013-03-23

    Strangles is the main upper respiratory tract disease of horses. There are currently no studies on the changes in alleles of the M protein gene (seM) in Brazilian isolates of Streptococcus equi ssp. equi (S. equi). This study aimed to analyze and differentiate molecularly S. equi isolates from equine clinical specimens from southern Brazil, between 1994 and 2010. seM alleles were analyzed in 47 isolates of S. equi obtained from clinical cases of strangles (15 Thoroughbred horses, 29 Crioulo breed horses and three Brasileiro de Hipismo--BH). seM alleles characterization was performed by comparing variable region sequences of the seM gene. The alleles were also phylogenetically grouped by Neighbor-joining analysis, which demonstrated the geographic distribution of those in properties from southern Brazil. Fifteen alleles of the gene seM were found among the 47 S. equi isolates analyzed. Among these, only one allele (seM-61), which was identified in seven isolates (14.9%), was found in the database PubMLST-seM. Within the new alleles, allele seM-115 was the most prevalent, having been found in 13 isolates (27.7%), followed by allele seM-117 in 10 isolates (21.3%). In the Brazilian horse population studied, there is greater diversity of M protein alleles in S. equi isolates compared to worldwide data deposited in PubMLST-seM. Among the 15 seM alleles identified, only one allele sequence was previously published. The alleles identification is important to control the disease by guiding selection of strains for the manufacture of commercial and autogenous vaccines. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. The Effectiveness of Anti-R. equi Hyperimmune Plasma against R. equi Challenge in Thoroughbred Arabian Foals of Mares Vaccinated with R. equi Vaccine

    PubMed Central

    Erganis, Osman; Sayin, Zafer; Hadimli, Hasan Huseyin; Sakmanoglu, Asli; Pinarkara, Yasemin; Ozdemir, Ozgur; Maden, Mehmet

    2014-01-01

    This study aimed to determine the effectiveness of a pregnant mare immunization of a Rhodococcus equi (R. equi) vaccine candidate containing a water-based nanoparticle mineral oil adjuvanted (Montanide IMS 3012) inactive bacterin and virulence-associated protein A (VapA), as well as the administration of anti-R. equi hyperimmune (HI) plasma against R. equi challenge in the mares' foals. The efficacy of passive immunizations (colostral passive immunity by mare vaccination and artificial passive immunity by HI plasma administration) was evaluated based on clinical signs, complete blood count, blood gas analysis, serological response (ELISA), interleukin-4 (IL-4) and interferon gamma (IFN-γ), total cell count of the bronchoalveolar lavage fluids (BALF) samples, reisolation rate of R. equi from BALF samples (CFU/mL), lung samples (CFU/gr), and lesion scores of the organs and tissue according to pathological findings after necropsy in the foals. The vaccination of pregnant mares and HI plasma administration in the foals reduced the severity of R. equi pneumonia and lesion scores of the organs and tissue by 3.54-fold compared to the control foals. This study thus indicates that immunization of pregnant mares with R. equi vaccine candidate and administration of HI plasma in mares' foals effectively protect foals against R. equi challenge. PMID:24982958

  1. The effectiveness of anti-R. equi hyperimmune plasma against R. equi challenge in thoroughbred Arabian foals of mares vaccinated with R. equi vaccine.

    PubMed

    Erganis, Osman; Sayin, Zafer; Hadimli, Hasan Huseyin; Sakmanoglu, Asli; Pinarkara, Yasemin; Ozdemir, Ozgur; Maden, Mehmet

    2014-01-01

    This study aimed to determine the effectiveness of a pregnant mare immunization of a Rhodococcus equi (R. equi) vaccine candidate containing a water-based nanoparticle mineral oil adjuvanted (Montanide IMS 3012) inactive bacterin and virulence-associated protein A (VapA), as well as the administration of anti-R. equi hyperimmune (HI) plasma against R. equi challenge in the mares' foals. The efficacy of passive immunizations (colostral passive immunity by mare vaccination and artificial passive immunity by HI plasma administration) was evaluated based on clinical signs, complete blood count, blood gas analysis, serological response (ELISA), interleukin-4 (IL-4) and interferon gamma (IFN- γ ), total cell count of the bronchoalveolar lavage fluids (BALF) samples, reisolation rate of R. equi from BALF samples (CFU/mL), lung samples (CFU/gr), and lesion scores of the organs and tissue according to pathological findings after necropsy in the foals. The vaccination of pregnant mares and HI plasma administration in the foals reduced the severity of R. equi pneumonia and lesion scores of the organs and tissue by 3.54-fold compared to the control foals. This study thus indicates that immunization of pregnant mares with R. equi vaccine candidate and administration of HI plasma in mares' foals effectively protect foals against R. equi challenge.

  2. Genotypic and phenotypic detection of efflux pump in Rhodococcus equi.

    PubMed

    Gressler, Letícia Trevisan; de Vargas, Agueda Castagna; da Costa, Mateus Matiuzzi; Pötter, Luciana; da Silveira, Bibiana Petri; Sangioni, Luis Antônio; de Avila Botton, Sônia

    2014-01-01

    The req_39680 gene, associated to a putative efflux system, was detected in 60% (54/90) of R. equi isolates by PCR. The phenotypic expression of efflux mechanism was verified in 20% of the isolates using ethidium bromide. For the first time, the expression of efflux mechanism was demonstrated in R. equi.

  3. Activity of 10 antimicrobial agents against intracellular Rhodococcus equi.

    PubMed

    Giguère, Steeve; Berghaus, Londa J; Lee, Elise A

    2015-08-05

    Studies with facultative intracellular bacterial pathogens have shown that evaluation of the bactericidal activity of antimicrobial agents against intracellular bacteria is more closely associated with in vivo efficacy than traditional in vitro susceptibility testing. The objective of this study was to determine the relative activity of 10 antimicrobial agents against intracellular Rhodococcus equi. Equine monocyte-derived macrophages were infected with virulent R. equi and exposed to erythromycin, clarithromycin, azithromycin, rifampin, ceftiofur, gentamicin, enrofloxacin, vancomycin, imipenem, or doxycycline at concentrations achievable in plasma at clinically recommended dosages in foals. The number of intracellular R. equi was determined 48h after infection by counting colony forming units (CFUs). The number of R. equi CFUs in untreated control wells were significantly higher than those of monolayers treated with antimicrobial agents. Numbers of R. equi were significantly lower in monolayers treated with enrofloxacin followed by those treated with gentamicin, and vancomycin, when compared to monolayers treated with other antimicrobial agents. Numbers of R. equi in monolayers treated with doxycycline were significantly higher than those of monolayers treated with other antimicrobial agents. Differences in R. equi CFUs between monolayers treated with other antimicrobial agents were not statistically significant. Enrofloxacin, gentamicin, and vancomycin are the most active drugs in equine monocyte-derived macrophages infected with R. equi. Additional studies will be needed to determine if these findings correlate with in vivo efficacy. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Phenotypic and genotypic characterization of Rhodococcus equi isolated from sputum.

    PubMed

    Silva, Paulo da; Santos, Adolfo Carlos Barreto; Sato, Daisy Nakamura; Silva, Jaqueline Otero; Medeiros, Marta Inês Cazentini; Carneiro, Ana Maria Machado; Leite, Sergio Roberto de Andrade; Leite, Clarice Queico Fujimura

    2012-01-01

    Rhodococcus equi is an opportunistic pathogen, causing rhodococcosis, a condition that can be confused with tuberculosis. Often, without identifying M. tuberculosis, physicians initiate empiric treatment for tuberculosis. R. equi and M. tuberculosis have different susceptibility to drugs. Identification of R. equi is based on a variety of phenotypic, chromatographic, and genotypic characteristics. This study aimed to characterize bacterial isolates from sputum samples suggestive of R. equi. The phenotypic identification included biochemical assays; thin-layer chromatography (TLC) and polymerase chain reaction (PCR) were used for genotypic identification. Among 78 Gram-positive and partially acid-fast bacilli isolated from the sputum of tuberculosis-suspected patients, 51 were phenotypically and genotypically characterized as R. equi based on literature data. Mycolic acid analysis showed that all suspected R. equi had compounds with a retention factor (R(f)) between 0.4-0.5. Genotypic characterization indicated the presence of the choE gene 959bp fragments in 51 isolates CAMP test positive. Twenty-two CAMP test negative isolates were negative for the choE gene. Five isolates presumptively identified as R. equi, CAMP test positive, were choE gene negative, and probably belonged to other bacterial species. The phenotypic and molecular techniques used constitute a good methodological tool to identify R. equi. Copyright © 2012 Elsevier Editora Ltda. All rights reserved.

  5. Pangenome and Phylogenomic Analysis of the Pathogenic Actinobacterium Rhodococcus equi

    PubMed Central

    Anastasi, Elisa; MacArthur, Iain; Scortti, Mariela; Alvarez, Sonsiray; Giguère, Steeve; Vázquez-Boland, José A.

    2016-01-01

    We report a comparative study of 29 representative genomes of the animal pathogen Rhodococcus equi. The analyses showed that R. equi is genetically homogeneous and clonal, with a large core genome accounting for ≈80% of an isolates’ gene content. An open pangenome, even distribution of accessory genes among the isolates, and absence of significant core–genome recombination, indicated that gene gain/loss is a main driver of R. equi genome evolution. Traits previously predicted to be important in R. equi physiology, virulence and niche adaptation were part of the core genome. This included the lack of a phosphoenolpyruvate:carbohydrate transport system (PTS), unique among the rhodococci except for the closely related Rhodococcus defluvii, reflecting selective PTS gene loss in the R. equi–R. defluvii sublineage. Thought to be asaccharolytic, rbsCB and glcP non-PTS sugar permease homologues were identified in the core genome and, albeit inefficiently, R. equi utilized their putative substrates, ribose and (irregularly) glucose. There was no correlation between R. equi whole-genome phylogeny and host or geographical source, with evidence of global spread of genomovars. The distribution of host-associated virulence plasmid types was consistent with the exchange of the plasmids (and corresponding host shifts) across the R. equi population, and human infection being zoonotically acquired. Phylogenomic analyses demonstrated that R. equi occupies a central position in the Rhodococcus phylogeny, not supporting the recently proposed transfer of the species to a new genus. PMID:27638249

  6. Comparative genomic analysis and phylogenetic position of Theileria equi

    USDA-ARS?s Scientific Manuscript database

    Transmission of arthropod-borne apicomplexan parasites that cause disease and result in death or persistent infection represents a major challenge to global human and animal health. First described in 1901 as Piroplasma equi, this re-emergent apicomplexan parasite was renamed Babesia equi and subseq...

  7. Genotypic and phenotypic detection of efflux pump in Rhodococcus equi

    PubMed Central

    Gressler, Letícia Trevisan; de Vargas, Agueda Castagna; da Costa, Mateus Matiuzzi; Pötter, Luciana; da Silveira, Bibiana Petri; Sangioni, Luis Antônio; de Avila Botton, Sônia

    2014-01-01

    The req_39680 gene, associated to a putative efflux system, was detected in 60% (54/90) of R. equi isolates by PCR. The phenotypic expression of efflux mechanism was verified in 20% of the isolates using ethidium bromide. For the first time, the expression of efflux mechanism was demonstrated in R. equi. PMID:25242956

  8. EquiX-A Search and Query Language for XML.

    ERIC Educational Resources Information Center

    Cohen, Sara; Kanza, Yaron; Kogan, Yakov; Sagiv, Yehoshua; Nutt, Werner; Serebrenik, Alexander

    2002-01-01

    Describes EquiX, a search language for XML that combines querying with searching to query the data and the meta-data content of Web pages. Topics include search engines; a data model for XML documents; search query syntax; search query semantics; an algorithm for evaluating a query on a document; and indexing EquiX queries. (LRW)

  9. Klossiella equi in the kidneys of a horse.

    PubMed

    Austin, R J; Dies, K H

    1981-05-01

    The protozoan, Klossiella equi was found in the kidneys of an aged Shetland mare raised in the Fredericton area of New Brunswick. This is the first published report of K. equi in a horse in Canada. The microscopic appearance of the parasite in the kidney is described. A brief discussion of other conditions seen in the horse is also presented.

  10. EquiX-A Search and Query Language for XML.

    ERIC Educational Resources Information Center

    Cohen, Sara; Kanza, Yaron; Kogan, Yakov; Sagiv, Yehoshua; Nutt, Werner; Serebrenik, Alexander

    2002-01-01

    Describes EquiX, a search language for XML that combines querying with searching to query the data and the meta-data content of Web pages. Topics include search engines; a data model for XML documents; search query syntax; search query semantics; an algorithm for evaluating a query on a document; and indexing EquiX queries. (LRW)

  11. Streptococcus equi subspecies equi in horses in Israel: seroprevalence and strain types

    PubMed Central

    Tirosh-Levy, S.; Blum, S. E.; Steward, K. F.; Waller, A. S.; Steinman, A.

    2016-01-01

    The purpose of this cross-sectional study was to determine the seroprevalence of Streptococcus equi in Israel, to monitor seropositive horses over time and to identify archived strains that were recovered from Israeli horses. A serological survey of 200 healthy horses on 20 farms throughout Israel was performed to detect recent exposure to S equi antigens A and C via indirect ELISA. Seroprevalence was 9.5 per cent (19/200) and positive horses were found in 30 per cent (6/20) of the farms. Sixteen horses that returned a positive serology result were retested three and six months later. Most (12/16) positive horses remained positive, which suggests the presence of animals with persistent infection. Molecular characterisation of S equi strains by sequencing of the SeM gene of 16 archived isolates of S equi that were recovered from clinical cases of strangles between 2008 and 2012 identified two strains: SeM-2 and SeM-28. PMID:27651915

  12. Comparative genomic analyses reveal a lack of a substantial signature of host adaptation in Rhodococcus equi ('Prescottella equi').

    PubMed

    Sangal, Vartul; Jones, Amanda L; Goodfellow, Michael; Sutcliffe, Iain C; Hoskisson, Paul A

    2014-08-01

    Rhodococcus equi ('Prescottella equi') is a pathogenic actinomycete primarily infecting horses but has emerged as an opportunistic human pathogen. We have sequenced the genome of the type strain of this species, R. equi strain C7(T) , and compared the genome with that of another foal isolate 103S and of a human isolate ATCC 33707. The R. equi strains are closely related to each other and yet distantly related to other rhodococci and Nocardia brasiliensis. The comparison of gene contents among R. equi strains revealed minor differences that could be associated with host adaptation from foals to humans, including the presence of a paa operon in the human isolate, which is potentially involved in pathogenesis. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  13. Molecular characterization of Rhodococcus equi isolates in equines

    PubMed Central

    Javed, Rabyia; Taku, A. K.; Sharma, R. K.; Badroo, Gulzaar Ahmed

    2017-01-01

    Aim: The aim was to determine the occurrence of Rhodococcus equi in equines and their environment in Jammu (R.S. Pura, Katra), molecular characterization and to determine the antibiotic resistance pattern of R. equi. Materials and Methods: A total of 96 nasopharyngeal swab samples were collected from equines. The organism was isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and was later confirmed by cultural characteristics and biochemical tests. Molecular detection of R. equi isolates was done by 16S rRNA gene amplification followed by virulence associated protein A (Vap A) gene amplification. Antibiogram was performed against five antibiotics, viz., amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. Results: During the study, 9 R. equi isolates were identified on the basis of cultural and biochemical tests. In the polymerase chain reaction based detection, 3 among the 9 rhodococcal isolates were positive for species-specific 16S rRNA gene and revealed amplicon of 450 bp for confirmation of 16S rRNA gene. None of the sample was found positive for Vap A gene. In antibiogram, R. equi isolates were found sensitive for amoxicillin, while some isolates were also found resistant to the most conventional antibiotic penicillin G. Conclusion: From this study, it was concluded that R. equi infection is prevalent in equines in Jammu region of India and the indiscriminate use of the antibiotics is leading toward the development of resistant strains of R. equi. PMID:28246441

  14. Molecular characterization of Rhodococcus equi isolates in equines.

    PubMed

    Javed, Rabyia; Taku, A K; Sharma, R K; Badroo, Gulzaar Ahmed

    2017-01-01

    The aim was to determine the occurrence of Rhodococcus equi in equines and their environment in Jammu (R.S. Pura, Katra), molecular characterization and to determine the antibiotic resistance pattern of R. equi. A total of 96 nasopharyngeal swab samples were collected from equines. The organism was isolated on Columbia nalidixic acid agar containing 5% sheep blood as well as on sheep blood agar and was later confirmed by cultural characteristics and biochemical tests. Molecular detection of R. equi isolates was done by 16S rRNA gene amplification followed by virulence associated protein A (Vap A) gene amplification. Antibiogram was performed against five antibiotics, viz., amoxicillin, penicillin G, streptomycin, rifampicin, and methicillin. During the study, 9 R. equi isolates were identified on the basis of cultural and biochemical tests. In the polymerase chain reaction based detection, 3 among the 9 rhodococcal isolates were positive for species-specific 16S rRNA gene and revealed amplicon of 450 bp for confirmation of 16S rRNA gene. None of the sample was found positive for Vap A gene. In antibiogram, R. equi isolates were found sensitive for amoxicillin, while some isolates were also found resistant to the most conventional antibiotic penicillin G. From this study, it was concluded that R. equi infection is prevalent in equines in Jammu region of India and the indiscriminate use of the antibiotics is leading toward the development of resistant strains of R. equi.

  15. Disseminated Rhodococcus equi infection in dromedary camels (Camelus dromedarius).

    PubMed

    Kinne, J; Madarame, H; Takai, S; Jose, S; Wernery, U

    2011-04-21

    Rhodococcus (R). equi, a recognized pathogen in horses, is emerging as a human opportunistic pathogen, especially in immunocompromized people. It affects also New World camelids, but there are no reports of R. equi infection in Old World camelids yet. Four cases of disseminated R. equi infection in adult breeding dromedaries occurred at one camel farm near Dubai within 16 months of each other. At necropsy the lungs were diffusely consolidated with large caseous areas. Histology revealed severe suppurative to necrotising pneumonia with multiple encapsulated abscesses. Immunohistochemistry enabled the detection of 15- to 17-kDa antigens (VapA) of R. equi in the lung sections. High numbers of R. equi were isolated from the lung lesions as well as from liver, spleen and mediastinal lymph nodes, indicative of septicaemia. The isolated strains were PCR-positive for the specific virulence plasmid (VapA-Gen) of R. equi, indicating virulent strains and containing an 85-kb type I plasmid. This is the first report of disseminated R. equi infection in Old World camelids. Since adult camels in general do not suffer from bacterial caused pneumonia (except tuberculosis), this is a new emerging disease for camels. Copyright © 2010 Elsevier B.V. All rights reserved.

  16. Pangenome and Phylogenomic Analysis of the Pathogenic Actinobacterium Rhodococcus equi.

    PubMed

    Anastasi, Elisa; MacArthur, Iain; Scortti, Mariela; Alvarez, Sonsiray; Giguère, Steeve; Vázquez-Boland, José A

    2016-10-23

    We report a comparative study of 29 representative genomes of the animal pathogen Rhodococcus equi The analyses showed that R. equi is genetically homogeneous and clonal, with a large core genome accounting for ≈80% of an isolates' gene content. An open pangenome, even distribution of accessory genes among the isolates, and absence of significant core-genome recombination, indicated that gene gain/loss is a main driver of R. equi genome evolution. Traits previously predicted to be important in R. equi physiology, virulence and niche adaptation were part of the core genome. This included the lack of a phosphoenolpyruvate:carbohydrate transport system (PTS), unique among the rhodococci except for the closely related Rhodococcus defluvii, reflecting selective PTS gene loss in the R. equi-R. defluvii sublineage. Thought to be asaccharolytic, rbsCB and glcP non-PTS sugar permease homologues were identified in the core genome and, albeit inefficiently, R. equi utilized their putative substrates, ribose and (irregularly) glucose. There was no correlation between R. equi whole-genome phylogeny and host or geographical source, with evidence of global spread of genomovars. The distribution of host-associated virulence plasmid types was consistent with the exchange of the plasmids (and corresponding host shifts) across the R. equi population, and human infection being zoonotically acquired. Phylogenomic analyses demonstrated that R. equi occupies a central position in the Rhodococcus phylogeny, not supporting the recently proposed transfer of the species to a new genus. © The Author(s) 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  17. An Adenoviral Vector Based Vaccine for Rhodococcus equi

    PubMed Central

    Giles, Carla; Ndi, Olasumbo; Barton, Mary D.; Vanniasinkam, Thiru

    2016-01-01

    Rhodococcus equi is a respiratory pathogen which primarily infects foals and is endemic on farms around the world with 50% mortality and 80% morbidity in affected foals. Unless detected early and treated appropriately the disease can be fatal. Currently, there is no vaccine available to prevent this disease. For decades researchers have endeavoured to develop an effective vaccine to no avail. In this study a novel human adenoviral vector vaccine for R. equi was developed and tested in the mouse model. This vaccine generated a strong antibody and cytokine response and clearance of R. equi was demonstrated following challenge. These results show that this vaccine could potentially be developed further for use as a vaccine to prevent R. equi disease in foals. PMID:27008624

  18. [Periprosthetic joint infection caused by Rhodococcus equi. Case report].

    PubMed

    Sallai, Imre; Péterfy, Nóra; Sanatkhani, Mohammad; Bejek, Zoltán; Antal, Imre; Prinz, Gyula; Kristóf, Katalin; Skaliczki, Gábor

    2017-07-01

    Rhodococcus equi is a rare pathogen in humans causing infections mostly in immunocompromised hosts. We present the first case of periprosthetic joint infection caused by Rhodococcus equi. An 88-year-old male patient was referred to our clinic with a history of fever and right hip pain. The patient had multiple hip surgeries including total joint arthroplasty and revision for aseptic loosening on the right side. He was immunocompetent, but his additional medical history was remarkable for diabetes mellitus, diabetic nephropathy and stroke with hemiplegia resulting in immobilization. Radiography showed stable components, joint aspirate yielded Rhodococcus equi. Irrigation and debridement was proposed, but the patient refused any surgical intervention. Therefore antibiotic therapy was administered. At the last follow-up the patient is free of complaints but the C-reactive protein level is still elevated. This case illustrates the possible role of Rhodococcus equi in medical device-associated infections. Orv Hetil. 2017; 158(27): 1071-1074.

  19. An Adenoviral Vector Based Vaccine for Rhodococcus equi.

    PubMed

    Giles, Carla; Ndi, Olasumbo; Barton, Mary D; Vanniasinkam, Thiru

    2016-01-01

    Rhodococcus equi is a respiratory pathogen which primarily infects foals and is endemic on farms around the world with 50% mortality and 80% morbidity in affected foals. Unless detected early and treated appropriately the disease can be fatal. Currently, there is no vaccine available to prevent this disease. For decades researchers have endeavoured to develop an effective vaccine to no avail. In this study a novel human adenoviral vector vaccine for R. equi was developed and tested in the mouse model. This vaccine generated a strong antibody and cytokine response and clearance of R. equi was demonstrated following challenge. These results show that this vaccine could potentially be developed further for use as a vaccine to prevent R. equi disease in foals.

  20. Susceptibility testing of Rhodococcus equi: An interlaboratory test.

    PubMed

    Riesenberg, Anne; Kaspar, Heike; Feßler, Andrea T; Werckenthin, Christiane; Schwarz, Stefan

    2016-10-15

    Due to the importance of antimicrobial susceptibility testing (AST) for veterinary diagnostics, a standardised protocol for AST of Rhodococcus equi by broth microdilution has recently been developed and approved by the Clinical and Laboratory Standards Institute (CLSI). The aim of the present study was to test this protocol in an interlaboratory comparative study for its fitness for use in routine laboratory diagnostics. All of the 18 participating laboratories determined the minimum inhibitory concentrations (MIC) of two R. equi strains against 24 antimicrobial agents. The modal MIC values were determined and the acceptable ranges were set as the modal MIC ±1 dilution step. The R. equi field strain Rh110 showed a slightly better performance than the type strain R. equi ATCC(®) 25729. For the different antimicrobial agents tested, the percentage of MIC values within the acceptable ranges varied from 75.9 to 100% for R. equi ATCC(®) 25729, and from 85.2 to 100% for R. equi Rh110. The most homogeneous MIC results (i.e. modal MIC ±1 dilution step) were obtained for oxacillin and vancomycin, while the most divergent results were seen with cefotaxime and ceftiofur. Using a success rate of at least 80% of the strain-specific MICs being within the acceptable ranges as an arbitrary cut-off, only one of the participating laboratories failed to reach this cut-off value for one of the two R. equi strains. Thus, we consider the new protocol fit for use in routine AST of R. equi. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Streptococcus zooepidemicus and Streptococcus equi evolution: the role of CRISPRs.

    PubMed

    Waller, Andrew S; Robinson, Carl

    2013-12-01

    The host-restricted bacterium Streptococcus equi is the causative agent of equine strangles, the most frequently diagnosed infectious disease of horses worldwide. The disease is characterized by abscessation of the lymph nodes of the head and neck, leading to significant welfare and economic cost. S. equi is believed to have evolved from an ancestral strain of Streptococcus zooepidemicus, an opportunistic pathogen of horses and other animals. Comparison of the genome of S. equi strain 4047 with those of S. zooepidemicus identified examples of gene loss due to mutation and deletion, and gene gain through the acquisition of mobile genetic elements that have probably shaped the pathogenic specialization of S. equi. In particular, deletion of the CRISPR (clustered regularly interspaced short palindromic repeats) locus in the ancestor of S. equi may have predisposed the bacterium to acquire and incorporate new genetic material into its genome. These include four prophages and a novel integrative conjugative element. The virulence cargo carried by these mobile genetic elements is believed to have shaped the ability of S. equi to cause strangles. Further sequencing of S. zooepidemicus has highlighted the diversity of this opportunistic pathogen. Again, CRISPRs are postulated to influence evolution, balancing the need for gene gain over genome stability. Analysis of spacer sequences suggest that these pathogens may be susceptible to a limited range of phages and provide further evidence of cross-species exchange of genetic material among Streptococcus pyogenes, Streptococcus agalactiae and Streptococcus dysgalactiae.

  2. Survival and replication of Rhodococcus equi in macrophages.

    PubMed Central

    Hondalus, M K; Mosser, D M

    1994-01-01

    Rhodococcus equi is a facultative intracellular bacterium of macrophages that can cause serious pneumonia in both young horses and immunocompromised people. Essential to understanding rhodococcus pathogenesis is a quantitative documentation of the intracellular events that follow macrophage phagocytosis of the organism. By using a bacterial immunofluorescence staining assay, we verified the intracellular survival and replicative potential of R. equi in both murine peritoneal macrophages and equine alveolar macrophages in vitro. Following an initial lag period of 6 to 12 h, the intracellular numbers of R. equi begin to rise, often reaching macrophage-compromising levels by 48 h. A quantitative determination of bacterial growth by a novel image analysis cytometry technique confirmed our fluorescence microscopic results. By 48 h postinfection, bacterial numbers had increased by more than fivefold, and the majority of infected macrophages in the monolayer contained 10 or more bacteria per cell. The intracellular organisms were viable, as evidenced by the ability to incorporate radiolabeled uracil. The use of these techniques has identified differences in the in vitro replicative capacities of a virulent strain and an avirulent strain of R. equi. A clinical isolate of R. equi expressing a 17-kDa virulence-associated plasmid-encoded antigen was able to survive and replicate within macrophages, whereas an avirulent, non-plasmid-containing strain replicated poorly. These results suggest that plasmid-encoded bacterial virulence factors may contribute to the ability of R. equi to replicate within its host cell, the macrophage. Images PMID:7927672

  3. Use of a bacteriophage lysin for barnyard decontamination of Streptococcus equi

    USDA-ARS?s Scientific Manuscript database

    Equine strangles is a highly contagious purulent lymphadenitis of the head and neck that is caused by Streptococcus equi (S. equi subsp. equi). Acute swelling and subsequent abscess formation is found in the submaxillary, submandibular, and retropharyngeal lymph nodes causing a “strangling” of the ...

  4. Development and application of loop-mediated isothermal amplification methods targeting the seM gene for detection of Streptococcus equi subsp. equi.

    PubMed

    Hobo, Seiji; Niwa, Hidekazu; Oku, Kazuomi

    2012-03-01

    Loop-mediated isothermal amplification (LAMP) constitutes a potentially valuable diagnostic tool for rapid diagnosis of contagious diseases. In this study, we developed a novel LAMP method (seM-LAMP) to detect the seM gene of Streptococcus equi subsp. equi (S. equi), the causative agent of strangles in equids. The seM-LAMP successfully amplified the target sequence of the seM gene at 63°C within 60 min. The sensitivity of the seM-LAMP was slightly lower than the 2nd reaction of the seM semi-nested PCR. To evaluate the species specificity of the seM-LAMP, we tested 100 S. equi and 189 non-S. equi strains. Significant amplification of the DNA originating from S. equi was observed within 60 min incubation, but no amplification of non-S. equi DNA occurred. The results were identical to those of seM semi-nested PCR. To investigate the clinical usefulness of the methods, the seM-LAMP and the seM semi-nested PCR were used to screen 590 nasal swabs obtained during an outbreak of strangles. Both methods showed that 79 and 511 swabs were S. equi positive and negative, respectively, and the results were identical to those of the culture examination. These results indicate that the seM-LAMP is potentially useful for the reliable routine diagnosis of Streptococcus equi subsp. equi infections.

  5. Rhodococcus equi: the many facets of a pathogenic actinomycete.

    PubMed

    Vázquez-Boland, José A; Giguère, Steeve; Hapeshi, Alexia; MacArthur, Iain; Anastasi, Elisa; Valero-Rello, Ana

    2013-11-29

    Rhodococcus equi is a soil-dwelling pathogenic actinomycete that causes pulmonary and extrapulmonary pyogranulomatous infections in a variety of animal species and people. Young foals are particularly susceptible and develop a life-threatening pneumonic disease that is endemic at many horse-breeding farms worldwide. R. equi is a facultative intracellular parasite of macrophages that replicates within a modified phagocytic vacuole. Its pathogenicity depends on a virulence plasmid that promotes intracellular survival by preventing phagosome-lysosome fusion. Species-specific tropism of R. equi for horses, pigs and cattle appears to be determined by host-adapted virulence plasmid types. Molecular epidemiological studies of these plasmids suggest that human R. equi infection is zoonotic. Analysis of the recently determined R. equi genome sequence has identified additional virulence determinants on the bacterial chromosome. This review summarizes our current understanding of the clinical aspects, biology, pathogenesis and immunity of this fascinating microbe with plasmid-governed infectivity. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Transcriptional changes are involved in phenotype switching in Streptococcus equi subspecies equi.

    PubMed

    Steward, Karen F; Robinson, Carl; Waller, Andrew S

    2016-04-01

    Phenotypic heterogeneity within a population of bacteria, through genetic or transcriptional variation, enables survival and persistence in challenging and changing environments. We report here that a recent clinical isolate of S. equi, strain 1691 (Se1691), yielded a mixture of reduced capsule and mucoid colonies on primary isolation when grown on colistin-oxolinic acid blood agar (COBA) streptococcal selective plates. Passaging colonies of Se1691, with a reduced capsule phenotype maintained this mixed phenotype. In contrast, passaging mucoid colonies fixed the mucoid phenotype, suggesting adaptive genetic or transcriptional changes in response to growth on artificial media. However, despite obvious phenotypic and transcriptional differences, there were no apparent differences in the genome sequences of Se1691 recovered from colonies with a mucoid or reduced capsule phenotype. We identified 105 differentially transcribed genes in the transcriptomes of reduced capsule and mucoid colonies. The reduced capsule phenotype was associated with a significant reduction in transcription of the has locus (SEQ_0269 Q = 0.0015, SEQ_0270 Q = 0.0015, SEQ_0271 Q = 0.0285) and the amount of hyaluronic acid on the surface of S. equi recovered from non-mucoid colonies (P = 0.017). Significant differences in the transcription of 21 surface and secreted proteins were also observed. Our data show that changes in the bacterial transcriptome are linked to the mixed colony phenotype of Se1691.

  7. Disseminated Rhodococcus equi infection in a patient with Hodgkin lymphoma.

    PubMed

    Mikić, Dragan; Djordjević, Zoran; Sekulović, Leposava; Kojić, Miroslav; Tomanović, Branka

    2014-03-01

    Rhodococcus (R) equi is an opportunistic, uncommon human pathogen that causes mainly infection in immunocompromised hosts. The disease is usually presented as subacute pneumonia that is mostly cavitary and sometimes bacteremic. We reported the extremly rare case of a 43-year-old woman with Hodgkin lymphoma, who developed R equi pulmonary infection after recieving multiple courses of chemotherapy. Secondary, the patient developed bacteremia, leading to sepsis and dissemination of R equi infection in many extrapulmonary sites. At addmission the patient was febrile, tachypnoic, tachycardic, hypotensive, with fa cial edema, splenomegaly, positive meningeal signs, left hemiparesis and paraparesis. Laboratory data included erythrocyte sedimentation rate (ESR) > 140 mm/h, C-reactive protein (CRP) 143.0 mg/L, red blood cells (RBC) 2.14 x 10(12)/L, whyite blood cells (WBC) 2.8 x 10(9)/L, lactate dehydrogenase (LDH) 706 U/L, serum albumin 26 g/L, sodium 127 mmol/L and potassium 2.7 mmol/L. Blood culture and culture of sputum and empyema were positive for R equi. Imaging studies demonstrated a large right cavitary pneumonia and abscess, empyema, pericarditis, mediastinal and intra-abdominal lymphadenopathy, brain and psoas abscesses, osteomyelitis and spondylodiscitis. The patient recovered completely after a 12-month treatment with combinations of parenteral and oral antibiotics (meropenem, vancomycin, teicoplanin, ciprofloxacin, rifampicin, macrolides etc), including drainage of abscesses and empyema. Eight years after completition of the treatment the patient was without recurrence of R equi infection and lymphoma. Since the eradication od R equi is very difficult, it is very important to make the diagnosis and initiate appropriate antibiotic therapy as soon as possible.

  8. Oxyuris equi: lack of efficacy in treatment with macrocyclic lactones.

    PubMed

    Wolf, Denis; Hermosilla, Carlos; Taubert, Anja

    2014-03-17

    Whilst anthelminthic resistance of small strongyles is well documented, anthelmintic failures against infections with Oxyuris equi have scarcely been published so far. We describe two cases of equine oxyurosis and the anthelminthic failure of macrocyclic lactones (moxidectin, ivermectin) resulting in persistent O. equi infections with continuous egg shedding. The horses were kept in two different herds in the federal state of Hessia, Germany. Herd A kept two geldings: an 8-year-old Welsh-Cob-Mix and a 7-year-old Haflinger. Herd B was composed of four animals: 2 Connemara-mares, 31 and 19 years old, one 18-year-old Connemara-gelding and a 27-year-old Norwegian Fjord mare. All animals had a case history of various anthelmintic treatments with macrocyclic lactones (moxidectin and ivermectin alternating irregulary) in 2010 and 2011, nonetheless, they continued to shed O. equi nematodes and eggs. Animals were treated anew with moxidectin by members of the institute and were continuously monitored on a daily base by adhesive tape samples. Follow-up examinations for the reappearance of eggs were performed for 30 days in Herd A and 57 days in Herd B. In total, recurrence of O. equi egg shedding was detected in three out of six horses within 1-4 weeks after treatment. In both herds accompanying horses sharing the same stable and paddock remained negative for detection of O. equi-eggs or worms throughout the whole observation period. This is the first report in Europe showing inefficacy of commercial ivermectin compounds and furthermore the first report at all documenting ineffectiveness of moxidectin compounds in the treatment of O. equi-infections in horses indicating a possible development of resistance or confirming an existing incomplete oxyuricidal efficacy. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Unusual Extrapulmonary Rhodococcus Equi Infection in a Kidney Transplant Patient.

    PubMed

    Varotti, Giovanni; Barabani, Caterina; Dodi, Ferdinando; Bertocchi, Massimo; Mondello, Rosalia; Cupo, Pierpaolo; Santori, Gregorio; Palombo, Domenico; Fontana, Iris

    2016-12-01

    Rhodococcus equi is a well-recognized pathogen in veterinary medicine that can also affect immuno-compromised human subjects. The most common clinical features in humans include necrotizing pneumonia with subacute pulmonary disease, progressive cough, chest pain and fever. We report a case of a 49-year-old kidney transplant patient who developed a Rhodococcus equi infection characterized by multiple abscesses of the soft tissues and muscles without any respiratory manifestation. Combining specific antibiotic therapy and surgical management of the abscesses without immunosuppression discontinuation led to a complete recovery of both patient and graft.

  10. Molecular characterization of Streptococcus equi subspecies equi isolated from an Ethiopian camel by ribotyping and PCR-ribotyping.

    PubMed

    Sechi, L A; Roger, F; Diallo, A; Yigezu, L M; Zanetti, S; Fadda, G

    1999-10-01

    The aim of this study was to characterize a S. equi subspecies equi strain isolated from an Ethiopian camel by different molecular techniques (Ribotyping and PCR-Ribotyping). We compared the results obtained with those generated from two strains of the Pasteur Collection. The ribotyping showed the highest power of differentiation, distinguishing between the strains analyzed, whereas PCR-Ribotyping was able only to differentiate the camel isolate but not the strains from the Pasteur Collection. The application of this technique will be very useful to establish a clonal relationship among equine and camelids strains and help the prevention and cure of the equine and camel pathology.

  11. Control of Bovicola Equi (Phthiraptera: Trichodectidae) with Dimilin and Permethrin

    USDA-ARS?s Scientific Manuscript database

    Lice are wingless ectoparasitic insects that can irritate and injure their hosts and transmit pathogens. Horses and ponies can be infested with a chewing louse, Bovicola equi (Denny) (Phthiraptera: Trichodectidae) that irritates the animals, creates skin lesions, causes hair loss, and generally redu...

  12. Radiologic features of Rhodococcus equi pneumonia in AIDS.

    PubMed

    Muntaner, L; Leyes, M; Payeras, A; Herrera, M; Gutierrez, A

    1997-01-01

    This report outlines the radiological features observed in three cases of Rhodococcus equi (R. equi) pneumonia in AIDS (acquired immunodeficiency syndrome) and reviews another 45 radiological reports published of this emerging opportunistic pneumonia in Human Immunodeficiency Virus (HIV) infected patients. The clinical signs in our three patients consisted in a subacute onset of respiratory symptoms and fever. A low lymphocyte count (< 200 cells/mm3), pulmonary infiltrates, and pleural effusion was present in all three cases. Cavitary pneumonia was observed in two patients, and pericardial effusion in another. In this series CD4 lymphocyte count < 200/mm3 was seen in 29 of the 48 patients (60.4%). All 48 patients had abnormal findings on chest radiographs. Abnormalities involved the upper lobes in 26 of the 48 patients (55%). Cavitation was reported in 37 of the 48 cases (77%). R. equi pneumonia may not be as the paucity of case reports suggest. Consequently, a cavitary pneumonia in HIV infected patients with a low CD4 lymphocyte count (< 200 mm3) with a subacute onset, an upper lobe predilection, and/or a poor response to conventional antibiotic therapy should be considered as suspect of R. equi infection.

  13. Amblyomma cajennense is an intrastadial biological vector of theileria equi

    USDA-ARS?s Scientific Manuscript database

    Background: The apicomplexan hemoprotozoan parasite Theileria equi is one of the etiologic agents causing equine piroplasmosis (EP), a disease of horses and their relatives that are endemic throughout large parts of the world. Prior to 2009 the United States had been considered to be free of this pa...

  14. Equi-surjective systems of linear operators and applications

    NASA Astrophysics Data System (ADS)

    Luc, D. T.; Minh, N. B.

    2008-01-01

    In this paper we study a system of linear operators between finite-dimensional Euclidean spaces. Emphasis is made on unbounded systems and sufficient conditions are established for their equi-surjectivity. An application is presented in which a system of approximate Jacobian matrices is used to obtain a parametric interior mapping theorem. A multiplier rule for vector problems is also derived.

  15. Acute osteomyelitis caused by Rhodococcus equi in an immunocompetent child.

    PubMed

    Sistla, Sujatha; Karthikeyan, Sivasangeetha; Biswas, Rakhi; Parija, Subhash Chandra; Patro, Dilip Kumar

    2009-01-01

    Rhodococcus equi is an unusual pathogen causing infections mostly in immunocompromised patients, particularly in those with human immunodeficiency virus (HIV). It has rarely been reported to affect immunocompetent hosts, where it usually presents as an isolated extrapulmonary lesion. We report a case of osteomyelitis caused by this organism in an immunocompetent host.

  16. Clinical and microbiological features of bacteremia with Streptococcus equi.

    PubMed

    Trell, Kristina; Nilson, Bo; Petersson, Ann-Cathrine; Rasmussen, Magnus

    2017-02-01

    Streptococcus equi (SE) rarely causes human infections. We identified 18 SE isolates from blood cultures. The focus of infection was unknown (n = 5), arthritis (n = 3), catheter-related (n = 2), pneumonia (n = 2), or other (n = 6). There were no fatalities. Several patients had animal contacts but there were no indications of clonal outbreaks.

  17. Streptococcus equi subsp. zooepidemicus Infections Associated with Guinea Pigs

    PubMed Central

    Young, Andrea; Levine, Seth J.; Garvin, Joseph P.; Brown, Susan; Turner, Lauren; Fritzinger, Angela; Gertz, Robert E.; Murphy, Julia M.; Vogt, Marshall; Beall, Bernard

    2015-01-01

    Streptococcus equi subsp. zooepidemicus is a known zoonotic pathogen. In this public health investigation conducted in Virginia, USA, in 2013, we identified a probable family cluster of S. zooepidemicus cases linked epidemiologically and genetically to infected guinea pigs. S. zooepidemicus infections should be considered in patients who have severe clinical illness and report guinea pig exposure. PMID:25531424

  18. Studies on the immunogenicity of Streptococcus equi vaccines in foals.

    PubMed Central

    Srivastava, S K; Barnum, D A

    1985-01-01

    The ability of either formalin-treated or heat-inactivated whole Streptococcus equi cell vaccines or partially purified M-protein of S. equi to give rise to protective antibody levels was studied in Standardbred foals by serological means. Two commercial preparations, i.e. a beta-propiolactone killed whole S. equi cell bacterin and a cell-free extract of S. equi cells were included in the study. The mean passive hemagglutination antibody titers (10 X log2) in sera of foals given either four doses of formalin-treated whole cell vaccine or an initial dose of formalin-treated followed by three doses of heat-inactivated vaccine with or without levamisole were significantly higher two weeks after the final dose. These passive hemagglutination antibody titers were higher in foals given formalin-treated whole cell vaccine (6.7 +/- 1.5) than given commercial bacterin (4.5 +/- 2.1). The passive hemagglutination antibody titers in all the groups decreased at 12 to 16 weeks after fourth dose of the vaccine. Foals given a commercial cell-free extract did not show a significant increase in passive hemagglutination antibody titers even up to four weeks after third dose. A group of six pony foals immunized with partially-purified M protein showed mean passive hemagglutination antibody titers lower than those observed in foals given whole cell vaccines. In a challenge experiment with S. equi, two of six foals vaccinated with partially-purified M-protein and all three controls developed clinical disease. The passive hemagglutination antibody of vaccinated foals increased after challenge, while at 28 days postchallenge the passive hemagglutination antibody titers of vaccinates and recovered controls were similar. PMID:4075235

  19. Epidemiological survey of Corynebacterium equi infections on five Ontario horse farms.

    PubMed Central

    Prescott, J F; Travers, M; Yager-Johnson, J A

    1984-01-01

    Corynebacterium equi was cultured from manure or soil on five horse-breeding farms in Ontario at monthly intervals on three occasions during the summer of 1982. The organism was widespread. Contamination by C. equi of the loafing paddock and pasture areas was significantly greater in a farm established 30 years than in two established for four and six years and there was a significant correlation between the C. equi burden in stables, paddocks and pastures and the length of use of the five farms for horses. In all farms, numbers of C. equi in pasture soil exceeded numbers in fresh manure, suggesting that environmental multiplication of the organism might occur. A farm with an endemic C. equi pneumonia problem differed significantly from the other four farms, where disease was not endemic, in the larger number of C. equi isolated in the stable area. By contrast the farm with a C. equi pasture soil burden significantly heavier than on all other farms had no deaths due to C. equi pneumonia. There was a correlation (r = 0.78, p = 0.061) between the number of cases of C. equi pneumonia on the farms and numbers of C. equi in the area of the stables, but not on the paddocks or pastures. About two-thirds of randomly chosen isolates from the farms belonged to the three capsular serotypes most commonly found in pneumonic foals. PMID:6713248

  20. Safety and immunogenicity of a live-attenuated auxotrophic candidate vaccine against the intracellular pathogen Rhodococcus equi.

    PubMed

    Lopez, A M; Townsend, H G G; Allen, A L; Hondalus, M K

    2008-02-13

    Rhodococcus equi causes serious pneumonia in neonatal foals and is an opportunistic pathogen of people with compromised cellular immunity. No effective vaccine against R. equi disease in foals is available. We tested the safety and immunogenicity of a live, fully attenuated riboflavin auxotrophic candidate vaccine strain of R. equi (R. equi rib-). We demonstrated that R. equi rib- is immunogenic and capable of inducing IFN-gamma responses in immunocompetent BALB/c mice, yet it is safe even in an immunocompromised SCID mouse infection model. Moreover, it protects immunocompetent mice against virulent R. equi challenge. In foals, R. equi rib- was likewise safe and stimulated serum R. equi-specific immune responses. A preliminary immunization strategy did not afford protection against virulent R. equi challenge and therefore, optimization of the vaccine formulation and or vaccination protocol will be necessary.

  1. Oral Administration of Electron-Beam Inactivated Rhodococcus equi Failed to Protect Foals against Intrabronchial Infection with Live, Virulent R. equi.

    PubMed

    Rocha, Joana N; Cohen, Noah D; Bordin, Angela I; Brake, Courtney N; Giguère, Steeve; Coleman, Michelle C; Alaniz, Robert C; Lawhon, Sara D; Mwangi, Waithaka; Pillai, Suresh D

    2016-01-01

    There is currently no licensed vaccine that protects foals against Rhodococcus equi-induced pneumonia. Oral administration of live, virulent R. equi to neonatal foals has been demonstrated to protect against subsequent intrabronchial challenge with virulent R. equi. Electron beam (eBeam)-inactivated R. equi are structurally intact and have been demonstrated to be immunogenic when administered orally to neonatal foals. Thus, we investigated whether eBeam inactivated R. equi could protect foals against developing pneumonia after experimental infection with live, virulent R. equi. Foals (n = 8) were vaccinated by gavaging with eBeam-inactivated R. equi at ages 2, 7, and 14 days, or gavaged with equal volume of saline solution (n = 4), and subsequently infected intrabronchially with live, virulent R. equi at age 21 days. The proportion of vaccinated foals that developed pneumonia following challenge was similar among the vaccinated (7/8; 88%) and unvaccinated foals (3/4; 75%). This vaccination regimen did not appear to be strongly immunogenic in foals. Alternative dosing regimens or routes of administration need further investigation and may prove to be immunogenic and protective.

  2. Efficacy of a Parapoxvirus ovis-based immunomodulator against equine herpesvirus type 1 and Streptococcus equi equi infections in horses.

    PubMed

    Ons, Ellen; Van Brussel, Leen; Lane, Stephen; King, Vickie; Cullinane, Ann; Kenna, Rachel; Lyons, Pamela; Hammond, Toni-Ann; Salt, Jeremy; Raue, Rudiger

    2014-10-10

    The efficacy of Zylexis®, an immunomodulator in horses based on inactivated Parapoxvirus ovis (iPPVO), was assessed using an equine herpesvirus type 1 (EHV-1) challenge model in the presence of a natural infection with Streptococcus equi equi (S. equi). Eleven horses were treated with iPPVO and twelve were kept as controls. Six horses were challenged with EHV-1 and commingled with the horses on study. Animals were dosed on Days -2, 0 (just before commingling) and Day 7. On Day 11 significantly less nasal discharge, enlarged lymph nodes, EHV-1 shedding and lower rectal temperatures were observed in the iPPVO-treated group. In addition, iPPVO-treated horses showed significantly fewer enlarged lymph nodes on Days 17 and 19, significantly less lower jaw swelling on Day 3 and significantly lower rectal temperatures on Days 12 and 13. Dyspnoea, depression and anorexia were only recorded for the control group. Following challenge seven out of 11 horses in the iPPVO treated group shed EHV-1 but on Days 11, 12, 13, 14, 15 and 16 quantitative virus detection in this group was significantly lower as compared to the controls. All animals shed S. equi but the percentage of animals with positive bacterial detection was lower in the iPPVO group than in the control group from Day 14 through Day 28. This difference was significant on Day 24. No injection site reactions or adverse events were observed. In conclusion, Zylexis administration is safe and reduced clinical signs and shedding related to both EHV-1 and S. equi infections. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Survival of foals with experimentally induced Rhodococcus equi infection given either hyperimmune plasma containing R. equi antibody or normal equine plasma.

    PubMed

    Perkins, Gillian A; Yeager, Amy; Erb, Hollis N; Nydam, Daryl V; Divers, Thomas J; Bowman, James L

    2002-01-01

    The purpose of this study was to determine if colostrum-deprived foals with experimentally induced Rhodococcus equi pneumonia have a decreased severity of the disease and decreased mortality rate when given hyperimmune (HI) R. equi antibody plasma (R. equi titer at least 100 % and virulence-associated protein A [VapA] at least 10000) prophylactically versus when given normal equine plasma (R. equi titer less than 20 % and VapA less than 160). Sixteen colostrum-deprived foals (R. equi titer less than 5 %) each received normal equine plasma in the first 24 hours of life (R. equi titer less than 20 %). At 14 days of age, six foals were given normal equine plasma and 10 foals were given HI plasma. All foals were subsequently infected intrabronchially with a pathogenic strain of R. equi (2.5 x 10 sup 8; organisms) at 21 days of age. Repeated physical examinations, weight measurements, complete blood cell counts, fibrinogen measurements, and thoracic radiographs (ventrodorsal and lateral) were performed to help determine the severity of the disease. Foals given HI plasma had significantly higher R. equi ELISA titers (42.4 %) than those given normal plasma (20.9 %) on the day of experimental infection. Mortality rates and severity of disease were statistically similar (P >.05) for the groups. Although none of the foals was treated with antibiotics, several with severe R. equi pneumonia recovered. Either HI or normal equine plasma administered to foals in the first few weeks of life caused no adverse effects and may be protective against R. equi, although the exact constituent responsible for protection is undetermined and requires further investigation.

  4. Amblyomma cajennense is an intrastadial biological vector of Theileria equi

    PubMed Central

    2013-01-01

    Background The apicomplexan hemoprotozoan parasite Theileria equi is one of the etiologic agents causing equine piroplasmosis, a disease of equines that is endemic throughout large parts of the world. Before 2009 the United States had been considered to be free of this parasite. Occasional cases had occurred but there was no evidence for endemic vector-borne transmission in the U.S. until a 2009 outbreak in Texas in which Dermacentor variabilis and Amblyomma cajennense were implicated as vectors. Although D. variabilis has previously been shown to be a competent laboratory vector, studies suggested A. cajennense was not a competent transstadial vector, even though the presence of this tick species on horses in South American is epidemiologicaly correlated with higher a prevalence of infection. In this study we tested the transstadial and intrastadial vector competence of D. variabilis and A. cajennense for T. equi. Methods A tick passaged T. equi strain from the Texas outbreak and ticks colonized from engorged females collected off horses on the outbreak ranch in Texas were used for these studies. Nymph or adult ticks were fed on infected horses and transmission fed on naïve horses. Infections were tracked with PCR and serology, dissected tick tissues were tested with PCR. Results A. cajennense transmitted T. equi intrastadially when adult ticks acquired infection by feeding on an infected horse, and transmitted to a naïve host on subsequent reattachment and feeding. D. variabilis failed to transmit in the same experiment. Transstadial transmission was not successful for either tick species. PCR on DNA isolated from eggs of females that had fed on an infected horse suggests that there is no transovarial passage of this parasite by either tick species. Conclusion This work confirms that ticks from the Texas population of A. cajennense are competent intrastadial vectors of T. equi. We propose that the most likely natural mode of transmission for this parasite

  5. Activity of Clarithromycin or Rifampin Alone or in Combination against Experimental Rhodococcus equi Infection in Mice

    PubMed Central

    Burton, Alexandra J.; Berghaus, Londa J.; Hondalus, Mary K.

    2015-01-01

    Treatment of mice with the combination of clarithromycin with rifampin resulted in a significantly lower number of Rhodococcus equi CFU in the organs of mice than treatment with either drug alone or placebo. There was no significant difference in the number of R. equi CFU between mice treated with clarithromycin monotherapy, rifampin monotherapy, or placebo. The combination of clarithromycin with rifampin conferred a clear advantage over either drug as monotherapy in this model of chronic R. equi infection. PMID:25824218

  6. Mouse lung infection model to assess Rhodococcus equi virulence and vaccine protection.

    PubMed

    González-Iglesias, Patricia; Scortti, Mariela; MacArthur, Iain; Hapeshi, Alexia; Rodriguez, Héctor; Prescott, John F; Vazquez-Boland, José A

    2014-08-06

    The pathogenic actinomycete Rhodococcus equi causes severe purulent lung infections in foals and immunocompromised people. Although relatively unsusceptible to R. equi, mice are widely used for in vivo studies with this pathogen. The most commonly employed mouse model is based on systemic (intravenous) infection and determination of R. equi burdens in spleen and liver. Here, we investigated the murine lung for experimental infection studies with R. equi. Using a 10(7)CFU intranasal challenge in BALB/c mice, virulent R. equi consistently survived in quantifiable numbers up to 10 days in the lungs whereas virulence-deficient R. equi bacteria were rapidly cleared. An internally controlled virulence assay was developed in which the test R. equi strains are co-inoculated and monitored in the same mouse. Isogenic R. equi bacteria lacking either the plasmid vapA gene or the entire virulence plasmid were compared using this competitive assay. Both strains showed no significant differences in in vivo fitness in the lung, indicating that the single loss of the virulence factor VapA was sufficient to account for the full attenuation seen in the absence of the virulence plasmid. To test the adequacy of the lung infection model for monitoring R. equi vaccine efficacy, BALB/c mice were immunized with live R. equi and challenged intranasally. Vaccination conferred protection against acute pulmonary challenge with virulent R. equi. Our data indicate that the murine lung infection model provides a useful tool for both R. equi virulence and vaccine studies. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Immunogenicity of an electron beam inactivated Rhodococcus equi vaccine in neonatal foals.

    PubMed

    Bordin, Angela I; Pillai, Suresh D; Brake, Courtney; Bagley, Kaytee B; Bourquin, Jessica R; Coleman, Michelle; Oliveira, Fabiano N; Mwangi, Waithaka; McMurray, David N; Love, Charles C; Felippe, Maria Julia B; Cohen, Noah D

    2014-01-01

    Rhodococcus equi is an important pathogen of foals that causes severe pneumonia. To date, there is no licensed vaccine effective against R. equi pneumonia of foals. The objectives of our study were to develop an electron beam (eBeam) inactivated vaccine against R. equi and evaluate its immunogenicity. A dose of eBeam irradiation that inactivated replication of R. equi while maintaining outer cell wall integrity was identified. Enteral administration of eBeam inactivated R. equi increased interferon-γ production by peripheral blood mononuclear cells in response to stimulation with virulent R. equi and generated naso-pharyngeal R. equi-specific IgA in newborn foals. Our results indicate that eBeam irradiated R. equi administered enterally produce cell-mediated and upper respiratory mucosal immune responses, in the face of passively transferred maternal antibodies, similar to those produced in response to enteral administration of live organisms (a strategy which previously has been documented to protect foals against intrabronchial infection with virulent R. equi). No evidence of adverse effects was noted among vaccinated foals.

  8. Rhodococcus equi Sepsis in a Renal Transplant Recipient: A Case Study

    PubMed Central

    Macken, Eline; de Jonge, Hylke; Van Caesbroeck, Daniël; Verhaegen, Jan; Van Kerkhoven, Dana; Van Wijngaerden, Eric; Kuypers, Dirk

    2015-01-01

    Abstract Rhodococcus equi is an unusual cause of infection in humans, but has emerged as an opportunistic pathogen among immunocompromised patients. Primary pulmonary involvement is the most common clinical presentation, although the spectrum of disease is broad. Diagnosing R. equi infections remains challenging, both from clinical and microbiological view, and no standard treatment has been established. In this report, we present a detailed case of a 57-year-old male renal transplant recipient who developed R. equi bacteremia with a concomitant Pneumocystis jirovecii pneumonia. We describe the clinical features of R. equi infections, highlight the importance of an early diagnosis, and briefly review treatment options for this rare infection. PMID:27500216

  9. Immunogenicity of an Electron Beam Inactivated Rhodococcus equi Vaccine in Neonatal Foals

    PubMed Central

    Bordin, Angela I.; Pillai, Suresh D.; Brake, Courtney; Bagley, Kaytee B.; Bourquin, Jessica R.; Coleman, Michelle; Oliveira, Fabiano N.; Mwangi, Waithaka; McMurray, David N.; Love, Charles C.; Felippe, Maria Julia B.; Cohen, Noah D.

    2014-01-01

    Rhodococcus equi is an important pathogen of foals that causes severe pneumonia. To date, there is no licensed vaccine effective against R. equi pneumonia of foals. The objectives of our study were to develop an electron beam (eBeam) inactivated vaccine against R. equi and evaluate its immunogenicity. A dose of eBeam irradiation that inactivated replication of R. equi while maintaining outer cell wall integrity was identified. Enteral administration of eBeam inactivated R. equi increased interferon-γ production by peripheral blood mononuclear cells in response to stimulation with virulent R. equi and generated naso-pharyngeal R. equi-specific IgA in newborn foals. Our results indicate that eBeam irradiated R. equi administered enterally produce cell-mediated and upper respiratory mucosal immune responses, in the face of passively transferred maternal antibodies, similar to those produced in response to enteral administration of live organisms (a strategy which previously has been documented to protect foals against intrabronchial infection with virulent R. equi). No evidence of adverse effects was noted among vaccinated foals. PMID:25153708

  10. Characterization of the arginine deiminase of Streptococcus equi subsp. zooepidemicus.

    PubMed

    Hong, Kyongsu

    2006-09-01

    Streptococcus equi subsp. zooepidemicus is an important cause of infectious diseases in horses and rarely humans. Little is known about the virulence factors or protective antigens of S. equi subsp. zooepidemicus. In the present study, I designed original primers based on an alignment of the gene sagp(arcA) from Streptococcus pyogenes encoding streptococcal acid glycoprotein-arginine deiminase (SAGP/AD) to amplify the S. equi subsp. zooepidemicus counterpart sequence by polymerase chain reaction, and I analyzed the sagp(arcA) gene of the organism. Using chromosomal walking steps, I identified a contiguous eight-gene locus involved in SAGP/AD production. Their open reading frames were found to share significant homologies and to correspond closely in molecular mass to previously sequenced arc genes of S. pyogenes, thus they were designated ahrC.2 (arginine repressor), arcR (CRP/FNR transcription regulator), sagp(arcA) (streptococcal acid glycoprotein-arginine deiminase), putative acetyltransferase gene, arcB (ornithine carbamyl transferase), arcD (arginine-ornithine antiporter), arcT (Xaa-His peptidase), and arcC (carbamate kinase). The SAGP homologue of S. equi subsp. zooepidemicus (SzSAGP), encoded by arcA gene of the bacteria (arcA(SZ)), was successfully expressed in Escherichia coli and purified to homogeneity. When in vitro growth inhibitory activity of the recombinant SzSAGP was tested against MOLT-3 cells, it inhibited the growth of the cells during the 3 days of culture in a dose-dependent manner, accompanied by the induction of apoptotic cell death. The recombinant protein also possessed AD activity. By immunoblot analysis using both anti-SzSAGP-SfbI(H8) and anti-SfbI(H8) sera, I was able to demonstrate that the SzSAGP protein is expressed on the streptococcal surface.

  11. Oral Administration of Electron-Beam Inactivated Rhodococcus equi Failed to Protect Foals against Intrabronchial Infection with Live, Virulent R. equi

    PubMed Central

    Rocha, Joana N.; Cohen, Noah D.; Bordin, Angela I.; Brake, Courtney N.; Giguère, Steeve; Coleman, Michelle C.; Alaniz, Robert C.; Lawhon, Sara D.; Mwangi, Waithaka; Pillai, Suresh D.

    2016-01-01

    There is currently no licensed vaccine that protects foals against Rhodococcus equi–induced pneumonia. Oral administration of live, virulent R. equi to neonatal foals has been demonstrated to protect against subsequent intrabronchial challenge with virulent R. equi. Electron beam (eBeam)-inactivated R. equi are structurally intact and have been demonstrated to be immunogenic when administered orally to neonatal foals. Thus, we investigated whether eBeam inactivated R. equi could protect foals against developing pneumonia after experimental infection with live, virulent R. equi. Foals (n = 8) were vaccinated by gavaging with eBeam-inactivated R. equi at ages 2, 7, and 14 days, or gavaged with equal volume of saline solution (n = 4), and subsequently infected intrabronchially with live, virulent R. equi at age 21 days. The proportion of vaccinated foals that developed pneumonia following challenge was similar among the vaccinated (7/8; 88%) and unvaccinated foals (3/4; 75%). This vaccination regimen did not appear to be strongly immunogenic in foals. Alternative dosing regimens or routes of administration need further investigation and may prove to be immunogenic and protective. PMID:26828865

  12. Protective effects of passively transferred merozoite-specific antibodies against Theileria equi in horses with severe combined immunodeficiency

    USDA-ARS?s Scientific Manuscript database

    Theileria equi immune plasma was infused into young horses (foals) with severe combined immunodeficiency. Although all foals became infected following intravenous challenge with homologous T. equi merozoite stabilate, delayed time-to-peak parasitemia and enhanced survival occurred. Protective effect...

  13. Experimental Studies on the Pathogenesis of Corynebacterium equi Infection in Foals

    PubMed Central

    Prescott, J. F.; Johnson, J. A.; Markham, R. J. F.

    1980-01-01

    Four month-old foals were infected orally with 75 mL of a suspension of 5.0 × 108 Corynebacterium equi per mL. Two foals were killed after ten days and had scanty number of C. equi in the caeco-colic lymph nodes. No C. equi were recovered from the other two foals, killed 20 days after infection. No gross pathological change was detected in these four foals, although mild microscopic lesions were seen in the ileum of one foal. Results of lymphocyte blastogenesis using peripheral blood lymphocytes and C. equi antigens showed, however, that lymphocytes became sensitized to C. equi following this challenge. In a second experiment four month-old foals were given orally the same dose of organisms but on five consecutive days. Two foals were killed ten days after infection and showed mild histological changes in the large bowel mucosa and C. equi could be recovered from all intestinal lymph nodes cultured. In one of these foals moderate numbers of C. equi were present in the bronchial lymph node. Of the other two foals, one died after 22 days with severe ulcerative enterocolitis and intestinal lymphadenitis. Only one small pulmonary abscess was detected despite large numbers of C. equi in the lungs. The other foal developed similar intestinal changes and was euthanized 25 days after infection. No C. equi were detected in the lungs or bronchial lymph node. Lymphocyte blastogenesis in these animals showed a rapid rise in response to C. equi antigens. These studies suggest that C. equi pneumonia in foals does not always arise from an intestinal infection, that minor intestinal infection causes a cellular immune response and that massive exposure of the bowel over a sustained period is necessary to induce intestinal lesions. ImagesFig. 2.Fig. 3. PMID:7427776

  14. Combining two serological assays optimises sensitivity and specificity for the identification of Streptococcus equi subsp. equi exposure.

    PubMed

    Robinson, Carl; Steward, Karen F; Potts, Nicola; Barker, Colin; Hammond, Toni-ann; Pierce, Karen; Gunnarsson, Eggert; Svansson, Vilhjálmur; Slater, Josh; Newton, J Richard; Waller, Andrew S

    2013-08-01

    The detection of anti-Streptococcus equi antibodies in the blood serum of horses can assist with the identification of apparently healthy persistently infected carriers and the prevention of strangles outbreaks. The aim of the current study was to use genome sequencing data to develop an indirect enzyme linked immunosorbent assay (iELISA) that targets two S. equi-specific protein fragments. The sensitivity and specificity of the antigen A and antigen C iELISAs were compared to an SeM-based iELISA marketed by IDvet - diagnostic Vétérinaire (IDvet). Individually, each assay compromised specificity in order to achieve sufficient sensitivity (SeM iELISA had a sensitivity of 89.9%, but a specificity of only 77.0%) or sensitivity to achieve high specificity. However, combining the results of the antigen A and antigen C iELISAs permitted optimisation of both sensitivity (93.3%) and specificity (99.3%), providing a robust assay for the identification of horses exposed to S. equi.

  15. Specific immune responses are required to control parasitemia in Babesia equi infection.

    PubMed Central

    Knowles, D P; Kappmeyer, L S; Perryman, L E

    1994-01-01

    Horses possessing a normal immune system and spleen often control infection caused by Babesia equi. However, splenectomized horses are unable to control B. equi infection and usually succumb to the infection. To investigate the role of the spleen in the control of B. equi infection in the absence of specific immune responses, two 1-month-old foals with severe combined immunodeficiency (SCID) and two age-matched normal foals were inoculated with B. equi. The SCID foals became febrile seven days postinoculation and developed terminal parasitemias of 41 and 29%. The SCID foals had greater than 50% decreases in indices of total erythrocytes, packed-cell volumes, and hemoglobin concentrations. Both SCID foals were euthanized in extremis at 10 days postinoculation. As expected, the serum of the SCID foals lacked detectable antibodies to B. equi antigens. In contrast, the normal foals inoculated with B. equi produced detectable anti-erythrocyte-stage parasite antibodies by 7 days and controlled clinical disease by 12 days postinoculation. Although SCID foals lack functional T and B lymphocytes, they do possess complement, macrophages, granulocytes, and natural killer cells, as well as a spleen. Therefore, the data indicate that specific immune responses are required to control B. equi parasitemia but are not required for erythrocyte lysis in infected horses. Furthermore, the spleen is not able to control B. equi parasitemia in the absence of specific immune responses to parasite antigens. Images PMID:8168957

  16. Prevention of foal mortality due to Rhodococcus equi pneumonia on an endemically affected farm

    PubMed Central

    Prescott, John F.; Machang'u, Robert; Kwiecien, Jacek; Delaney, Kathy

    1989-01-01

    Over the course of one summer, foals on a horse breeding farm where Rhodococcus equi infection was endemic were examined clinically twice weekly for evidence of R. equi pneumonia. Examination usually commenced from the week of birth and continued for up to 14 weeks of age. Affected animals were treated with a variety of antimicrobial drugs and such treatment was often prolonged. For descriptive purposes, we regarded a foal as developing R. equi pneumonia if the rectal temperature rose above 39°C, the respiratory rate was over 40 per min, there were characteristic associated changes in respiratory sounds on auscultation, and the foal responded to antimicrobial treatment. In a group of 16 foals followed in this way, R. equi pneumonia developed in at least 14. Seven of these foals developed antibodies to equi factors as determined in a neutralizing antibody assay. Mean age of onset of these 14 foals was 3.4 weeks. Three foals developed the disease between 1.5-2.5 weeks of age, and 12 of 14 before four weeks of age. In previous years four to six foals died on the farm each summer of R. equi pneumonia; only one foal died of R. equi pneumonia on the farm during the summer of this study, and this foal did not form part of the study group. Early clinical recognition and treatment of R. equi pneumonia in foals on endemically affected farms may be an effective way to prevent deaths due to this infection. PMID:17423454

  17. Rhodococcus equi hyperimmune plasma decreases pneumonia severity after a randomised experimental challenge of neonatal foals.

    PubMed

    Sanz, M G; Loynachan, A; Horohov, D W

    2016-03-12

    Since a vaccine is not available against Rhodococcus equi, R equi-specific hyperimmune plasma (HIP) is commonly used, although its efficacy remains controversial. The objective of this study was to evaluate the ability of a commercially available HIP to prevent clinical rhodococcal pneumonia in neonatal foals after experimental challenge. British Veterinary Association.

  18. Molecular surveillance of Theileria equi and Anaplasma phagocytophilum infections in horses from Ukraine, Poland and Slovakia.

    PubMed

    Slivinska, Kateryna; Víchová, Bronislava; Werszko, Joanna; Szewczyk, Tomasz; Wróblewski, Zbigniew; Peťko, Branislav; Ragač, Ondrej; Demeshkant, Vitaliy; Karbowiak, Grzegorz

    2016-01-15

    A survey was undertaken to assess the prevalence of Theileria equi and Anaplasma phagocytophilum in some regions of Ukraine, Poland and Slovakia. Using a specific PCR assays, blood samples from 215 horses were tested. The prevalence of T. equi and A. phagocytophilum infection was 13.95% and 1.4%, respectively. BLAST analysis showed the isolates closest to the T. equi 18S rRNA and A. phagocytophilum msp4 gene sequences in GenBank with a similarity of ≥99%. No significant association was found between the T. equi PCR positivity and the age or sex of the horses. There was a significant association between the origin of horses and T. equi-PCR positivity. No significant association was found between the A. phagocytophilum-PCR positivity and the age, sex or origin. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Rhodococcus equi pericarditis in a patient living with HIV/AIDS.

    PubMed

    Gundelly, Praveen; Thornton, Alice; Greenberg, Richard N; McCormick, Malkanthie; Myint, Thein

    2014-01-01

    Rhodococcus equi, previously called Corynebacterium equi, is known to cause pneumonia in foals and swine. Although it was known to cause infection rarely in humans, R equi infection in humans has increased with the advent of HIV and increased use of immunosuppressants. We report a case of a 48-year-old male patient with newly diagnosed HIV/AIDS presenting with recurrent R equi bacteremia, pericardial effusion, and pericardial cyst. The infection was treated with drainage of the pericardial effusion and cyst and 2 weeks of intravenous vancomycin and 6 months of oral azithromycin and levofloxacin. Rhodococcus equi causes pericarditis and pericardial effusion. It can be effectively treated with debridement, drainage, and a prolonged course of antibiotics. In vitro antibiotic susceptibility should be checked as resistance to antibiotics can develop, especially if drainage is inadequate.

  20. Rhodococcus equi pneumonia in the foal--part 1: pathogenesis and epidemiology.

    PubMed

    Muscatello, Gary

    2012-04-01

    Rhodococcus equi pneumonia is a worldwide infectious disease of major concern to the equine breeding industry. The disease typically manifests in foals as pyogranulomatous bronchopneumonia, resulting in significant morbidity and mortality. Inhalation of aerosolised virulent R. equi from the environment and intracellular replication within alveolar macrophages are essential components of the pathogenesis of R. equi pneumonia in the foal. Recently documented evidence of airborne transmission between foals indicates the potential for an alternative contagious route of disease transmission. In the first of this two-part review, the complexity of the host, pathogen and environmental interactions that underpin R. equi pneumonia will be discussed through an exploration of current understanding of the epidemiology and pathogenesis of R. equi pneumonia in the foal. Copyright © 2011 Elsevier Ltd. All rights reserved.

  1. Streptococcus equi Detection Polymerase Chain Reaction Assay for Equine Nasopharyngeal and Guttural Pouch Wash Samples.

    PubMed

    Boyle, A G; Rankin, S C; Duffee, L; Boston, R C; Wheeler-Aceto, H

    2016-01-01

    Bacterial culture and polymerase chain reaction (PCR) assays for the detection of Streptococcus equi in nasopharyngeal washes (NPW) and guttural pouch lavage (GPL) samples have low sensitivity. In human diagnostics, processing of samples with flocked swabs has improved recovery rates of bacterial agents because of improved surface area and elution factors. For S. equi subsp. equi (S. equi) detection in NPW and GPL samples we hypothesized that: direct-PCR would be more reliable than flocked swab culture (FS culture); flocked swab PCR (FS-PCR) would be equivalent to direct-PCR; and FS culture would be more reliable than traditional culture. A total of 193 samples (134 NPW and 59 GPL) from 113 horses with either suspected S. equi infection, convalescing from a known S. equi infection, or asymptomatic horses screened for S. equi. Prospective study. Samples were submitted for S. equi direct-PCR. Using logistic regression, direct-PCR (gold standard) was compared to FS culture, traditional culture, and FS-PCR also performed. Direct-PCR was statistically more sensitive than FS-PCR, FS culture, and traditional culture (P < .001). All methods had sensitivities <70% relative to the direct-PCR. FS culture had a similar sensitivity relative to traditional culture. The odds of GPL samples being positive on direct-PCR (P = .030) and FS-PCR were greater than those for NPW samples (P = .021). Use of flocked swabs during laboratory preprocessing did not improve detection of S. equi via either PCR or bacterial culture from samples. Direct-PCR is the preferred method of detection of S. equi. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  2. Use of a bacteriophage lysin, PlyC, as an enzyme disinfectant against Streptococcus equi.

    PubMed

    Hoopes, J Todd; Stark, Caren J; Kim, Han Ah; Sussman, Daniel J; Donovan, David M; Nelson, Daniel C

    2009-03-01

    Streptococcus equi is the causative agent of the purulent infection equine strangles. This disease is transmitted through shedding of live bacteria from nasal secretions and abscess drainage or by contact with surfaces contaminated by the bacteria. Disinfectants are effective against S. equi, but inactivation by environmental factors, damage to equipment, and toxicity are of great concern. Bacteriophage-encoded lysins (cell wall hydrolases) have been investigated as therapeutic agents due to their ability to lyse susceptible gram-positive organisms. Here, we investigate the use of one lysin, PlyC, as a narrow-spectrum disinfectant against S. equi. This enzyme was active against >20 clinical isolates of S. equi, including both S. equi subsp. equi and S. equi subsp. zooepidemicus. Significantly, PlyC was 1,000 times more active on a per weight basis than Virkon-S, a common disinfecting agent, with 1 microg of enzyme able to sterilize a 10(8) CFU/ml culture of S. equi in 30 min. PlyC was subjected to a standard battery of tests including the Use Dilution Method for Testing Disinfectants and the Germicidal Spray Products Test. Results indicate that aerosolized PlyC can eradicate or significantly reduce the S. equi load on a variety of materials found on common stable and horse-related equipment. Additionally, PlyC was shown to retain full activity under conditions that mimic a horse stable, i.e., in the presence of nonionic detergents, hard water, or organic materials. We propose PlyC as the first protein-based, narrow-spectrum disinfectant against S. equi, which may augment or supplement the use of broad-spectrum disinfectants in barns and stables where equine strangles is prevalent.

  3. Sequence heterogeneity in the equi merozoite antigen gene (ema-1) of Theileria equi and development of an ema-1-specific TaqMan MGB assay for the detection of T. equi.

    PubMed

    Bhoora, Raksha; Quan, Melvyn; Matjila, Paul T; Zweygarth, Erich; Guthrie, Alan J; Collins, Nicola E

    2010-08-27

    Although a quantitative real-time PCR assay (qPCR) assay for the detection of Theileria equi has been developed and evaluated, it is possible that additional, as yet undetected 18S rRNA gene sequence variants may exist. A qPCR assay targeting a different gene, used in conjunction with the T. equi 18S rRNA qPCR assay, could assist in the detection of all T. equi genotypes in field samples. A T. equi ema-1-specific qPCR (Ueti et al., 2003) was tested on 107 South African field samples, 90 of which tested positive for T. equi antibody using the immuno-fluorescent antibody test (IFAT). The qPCR assay performed poorly, as T. equi was detected in only 67 of the 90 IFAT-positive field samples at quantification cycle (C(q)) values ranging from 27 to 39.95. Furthermore, a high C(q) value of 36.18 was obtained from DNA extracted from a South African in vitro-cultured T. equi WL isolate [1.38% parasitized erythrocytes (PE)] when a low C(q) value (indicative of a high T. equi DNA concentration) was expected. Approximately 600 bp of the ema-1 gene from 38 South African samples were sequenced and BLASTN analysis confirmed all sequences to be merozoite surface protein genes, with an identity of 87.1-100% to previously published T. equi ema-1 gene sequences. Alignment of the sequences revealed extensive sequence variations in the target regions of the primers and probes (Ueti et al., 2003), explaining the poor performance of the qPCR assay. Based on these observations, we developed a new TaqMan minor-groove binder (MGB) probe-based qPCR assay, targeting a more conserved region of the ema-1 gene. This assay was shown to be efficient and specific, and the detection limit, defined as the concentration at which 95% of T. equi-positive samples are detected, was determined to be 1.4 x 10(-4)% PE. The two ema-1 assays were compared by testing 41 South African field samples in parallel. The results suggested that the new assay was more sensitive than the original assay, as T. equi was

  4. Seroprevalence of Rhodococcus equi in horses in Israel.

    PubMed

    Tirosh-Levy, Sharon; Gürbilek, Sevil E; Tel, Osman Y; Keskin, Oktay; Steinman, Amir

    2017-06-26

    Rhodococcus equi is a common cause of pneumonia in foals and has extensive clinical, economic and possibly zoonotic consequences. This bacterium survives well in the environment and may be considered as normal flora of adult horses. Certain strains of this bacterium are extremely virulent in foals, and early identification and intervention is crucial for prognosis. Rhodococcus equi is endemic in many parts of the world and occasionally isolated in Israel. This study was designed to evaluate R. equi seroprevalence in adult horses in Israel to indirectly indicate the potential level of exposure of susceptible foals. Sera were collected from 144 horses during spring 2011 and from 293 horses during fall 2014, and the presence of antibodies against virulent R. equi was detected by enzyme-linked immunosorbent assay. Equine seroprevalence of R. equi was found to be 7.6% in 2011 and 5.1% in 2014. Only one farm had seropositive horses in 2011, whereas several farms had seropositive horses in 2014. No significant risk factors for seropositivity were found. Rhodococcus equi appears to be endemic in Israel. This is the first survey of R. equi in Israel that provides information on the epidemiology of this important bacterium.

  5. Theileria (Babesia) equi and Babesia caballi infections in horses in Galicia, Spain.

    PubMed

    Camacho, A T; Guitian, F J; Pallas, E; Gestal, J J; Olmeda, A S; Habela, M A; Telford, S R; Spielman, A

    2005-05-01

    The control of equine piroplasmosis is becoming increasingly important to maintain the international market open to the horse industry. The purpose of this study was to demonstrate the occurrence of equine piroplasmosis (Theileria equi and Babesia caballi) in Galicia, north-west Spain, and to compare haematological and serum biochemistry parameters between non-parasitaemic horses and horses parasitaemic with T. equi and B. caballi. Sixty serum samples (control group) were taken from healthy horses pastured on two farms, and examined for evidence of equine T. equi and B. caballi infection by indirect fluorescent antibody test (IFAT). Of the 60 samples, 24 (40%) and 17 (28.3%) samples were positive for T. equi and B. caballi, respectively. Twelve (20%) samples were positive for both parasites. Haematology and serum biochemistry were compared between controls and a series of 36 horses clinically affected by T. equi (25) or B. caballi (11). Compared with the healthy group, there was a 43% and 37% decrease in the haematocrit for T. equi and B. caballi infection, respectively. Parasitaemic horses presented an intense anaemia and serum biochemistry signs of liver damage. The anaemia was more severe in T. equi-infected than in B. caballi-infected horses. Our results suggest that equine piroplasmosis is widespread in the region and is a cause for concern.

  6. Diagnosis and prevalence of Theileria equi horses in western Mexico by nested PCR.

    PubMed

    Ayala-Valdovinos, Miguel Angel; Lemus-Flores, Clemente; Galindo-García, Jorge; Bañuelos-Pineda, Jacinto; Rodríguez-Carpena, Javier Germán; Sánchez-Chiprés, David; Duifhuis-Rivera, Theodor

    2017-02-01

    Theileria equi infection prevalence was calculated from 1000 blood samples obtained from apparently healthy horses in western Mexico. Samples were sent to the Animal Biotechnology Laboratory of the University of Guadalajara (Mexico) for T. equi diagnosis. Nested polymerase chain reaction (nPCR) was used as a diagnostic method to detect pathogen DNA. Using primers for the merozoite antigen-1 (EMA-1) gene, 19.70±2.47% of the horses (95% CI, 17.23-22.17%) tested positive for T. equi. There was no significant association between gender and T. equi infection. However, prevalence was higher among stabled horses (25.81%) than that among grazing horses (15.02%). The positivity rate was also higher among Quarter Horse (24.70%), Lusitano (35.90%), and Costa Rican Saddle Horse (47.37%) breeds than that among the other seven breeds investigated in this study. The percentage of T. equi infection was higher among adult horses (≥ 4years old, 25.05%) than that among colts and fillies (2-4years old, 15.48%), yearlings (1-2years old, 10.49%), and foals (<1year old, 10.34%). This is the first study of T. equi infection prevalence among horses in Mexico by nPCR . The results indicate that the equine piroplasmosis (EP) caused by T. equi is enzootic in western Mexico.

  7. Prevalence and molecular characterization of Theileria equi and Babesia caballi in jereed horses in Erzurum, Turkey.

    PubMed

    Guven, Esin; Avcioglu, Hamza; Deniz, Ahmet; Balkaya, İbrahim; Abay, Ugur; Yavuz, Şevki; Akyüz, Muzaffer

    2017-03-01

    Equine piroplasmosis (EP) is a hemoprotozoan tick-borne disease with worldwide distribution that is caused by Theileria equi and Babesia caballi. There are studies reporting the presence of equine piroplasmosis in Turkey but the situation in Erzurum is unknown. The aim of the current study was to determine the situation of equine piroplasmosis in jeered horses in Erzurum. Between April and August 2015, a total of 125 Arabian horse were examined and blood samples were collected. At the time of sampling, animals were also examined for tick infestations and clinical signs. Besides microscopic examination of Giemsastained blood smears, multiplex PCR performed with species specific primers partially amplifying the 18S rRNA gene of B. caballi and T. equi. During the microscopic examination of blood smears, T. equi piroplasms were found in 6 (4.8%) samples. In total, 11 (8.8%) T. equi DNA were detected with multiplex PCR. B. caballi piroplasms or DNA were not obtained. BLAST analysis of the sequenced T. equi samples (GenBank: KU921661-KU921667) indicated 98.8-100% identity to each other, and 100% similarity to T. equi isolates in South Africa, Iran, China, Sudan, India, Mongolia, Trinidad, Kenya, Spain, Serbia, Bosnia and Herzegovina and Turkey (Bursa). The results of our study indicate that T. equi occurs more frequently than B. caballi in the study area. To the authors' knowledge, this is the first report of the molecular detection of equine piroplasmosis in jeered horses in Erzurum, Turkey.

  8. Phenotypic mutants of the intracellular actinomycete Rhodococcus equi created by in vivo Himar1 transposon mutagenesis.

    PubMed

    Ashour, Joseph; Hondalus, Mary K

    2003-04-01

    Rhodococcus equi is a facultative intracellular opportunistic pathogen of immunocompromised people and a major cause of pneumonia in young horses. An effective live attenuated vaccine would be extremely useful in the prevention of R. equi disease in horses. Toward that end, we have developed an efficient transposon mutagenesis system that makes use of a Himar1 minitransposon delivered by a conditionally replicating plasmid for construction of R. equi mutants. We show that Himar1 transposition in R. equi is random and needs no apparent consensus sequence beyond the required TA dinucleotide. The diversity of the transposon library was demonstrated by the ease with which we were able to screen for auxotrophs and mutants with pigmentation and capsular phenotypes. One of the pigmentation mutants contained an insertion in a gene encoding phytoene desaturase, an enzyme of carotenoid biosynthesis, the pathway necessary for production of the characteristic salmon color of R. equi. We identified an auxotrophic mutant with a transposon insertion in the gene encoding a putative dual-functioning GTP cyclohydrolase II-3,4-dihydroxy-2-butanone-4-phosphate synthase, an enzyme essential for riboflavin biosynthesis. This mutant cannot grow in minimal medium in the absence of riboflavin supplementation. Experimental murine infection studies showed that, in contrast to wild-type R. equi, the riboflavin-requiring mutant is attenuated because it is unable to replicate in vivo. The mutagenesis methodology we have developed will allow the characterization of R. equi virulence mechanisms and the creation of other attenuated strains with vaccine potential.

  9. Phenotypic Mutants of the Intracellular Actinomycete Rhodococcus equi Created by In Vivo Himar1 Transposon Mutagenesis

    PubMed Central

    Ashour, Joseph; Hondalus, Mary K.

    2003-01-01

    Rhodococcus equi is a facultative intracellular opportunistic pathogen of immunocompromised people and a major cause of pneumonia in young horses. An effective live attenuated vaccine would be extremely useful in the prevention of R. equi disease in horses. Toward that end, we have developed an efficient transposon mutagenesis system that makes use of a Himar1 minitransposon delivered by a conditionally replicating plasmid for construction of R. equi mutants. We show that Himar1 transposition in R. equi is random and needs no apparent consensus sequence beyond the required TA dinucleotide. The diversity of the transposon library was demonstrated by the ease with which we were able to screen for auxotrophs and mutants with pigmentation and capsular phenotypes. One of the pigmentation mutants contained an insertion in a gene encoding phytoene desaturase, an enzyme of carotenoid biosynthesis, the pathway necessary for production of the characteristic salmon color of R. equi. We identified an auxotrophic mutant with a transposon insertion in the gene encoding a putative dual-functioning GTP cyclohydrolase II-3,4-dihydroxy-2-butanone-4-phosphate synthase, an enzyme essential for riboflavin biosynthesis. This mutant cannot grow in minimal medium in the absence of riboflavin supplementation. Experimental murine infection studies showed that, in contrast to wild-type R. equi, the riboflavin-requiring mutant is attenuated because it is unable to replicate in vivo. The mutagenesis methodology we have developed will allow the characterization of R. equi virulence mechanisms and the creation of other attenuated strains with vaccine potential. PMID:12670990

  10. Chloroquine inhibits Rhodococcus equi replication in murine and foal alveolar macrophages by iron-starvation.

    PubMed

    Gressler, Leticia T; Bordin, Angela I; McQueen, Cole M; Cohen, Noah D; de Vargas, Agueda Castagna

    2016-05-30

    Rhodococcus equi preferentially infects macrophages causing pyogranulomatous pneumonia in young foals. Both the vapA and rhbC genes are up-regulated in an iron (Fe)-deprived environment, such as that found within macrophages. Chloroquine (CQ) is a drug widely used against malaria that suppresses the intracellular availability of Fe in eukaryotic cells. The main objective of this study was to evaluate the ability of CQ to inhibit replication of virulent R. equi within murine (J774A.1) and foal alveolar macrophages (AMs) and to verify whether the mechanism of inhibition could be Fe-deprivation-dependent. CQ effect on R. equi extracellular survival and toxicity to J774A.1 were evaluated. R. equi survival within J774A.1 and foal AMs was evaluated under CQ (10 and 20μM), bovine saturated transferrin (bHTF), and bovine unsaturated transferrin (bATF) exposure. To explore the action mechanism of CQ, the superoxide anion production, the lysozyme activity, as well as the relative mRNA expression of vapA and rhbC were examined. CQ at≤20μM had no effect on R. equi extracellular multiplication and J774A.1 viability. Exposure to CQ significantly and markedly reduced survival of R. equi within J774A.1 and foal AMs. Treatment with bHTF did not reverse CQ effect on R. equi. Exposure to CQ did not affected superoxide anion production or lysozyme activity, however vapA and rhbC expression was significantly increased. Our results reinforce the hypothesis that intracellular availability of Fe is required for R. equi survival, and our initial hypothesis that CQ can limit replication of R. equi in J774A.1 and foal AMs, most likely by Fe starvation. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Identification of carriers of Streptococcus equi in a naturally infected herd.

    PubMed

    George, J L; Reif, J S; Shideler, R K; Small, C J; Ellis, R P; Snyder, S P; McChesney, A E

    1983-07-01

    During an outbreak of strangles in a population of research horses, 4 mares were identified as carriers of Streptococcus equi. Three of the mares had typical signs of strangles (severe regional lymphadenitis with or without rupture of abscessed lymph nodes). The 4th mare experienced episodes of serous to mucopurulent nasal discharge, but never had more than a mild degree of lymph node enlargement. Streptococcus equi was isolated from the abscessed lymph nodes and from nasopharyngeal swab specimens from the first 3 mares from 6 to 19 weeks after rupture of involved nodes. Streptococcus equi was isolated from the nasopharynx of the 4th mare on introduction into the herd and intermittently over the ensuing 6 months. During the 7th month, mare 4 was placed in isolation, where she continued to shed S equi for 4 more months. A complete physical examination during the 10th month, including radiography of the head and thorax, did not reveal any relevant abnormalities, but a pharyngeal swab specimen was culture-positive for S equi. This isolate was used to inoculate 2 yearling colts, which developed strangles and from which S equi was reisolated. Shedding of S equi by mare 4 ceased in the 11th month, and at necropsy 2 months later, S equi was not recovered from any organ or tissue. Corticosteroid administration 3 weeks prior to necropsy had induced neither shedding of the organism nor clinical signs of strangles. The study provided clinical, epidemiologic, and bacteriologic evidence to support the existence of a carrier state following natural infection with S equi.

  12. Acute phase proteins in Andalusian horses infected with Theileria equi.

    PubMed

    Rodríguez, Rocío; Cerón, José J; Riber, Cristina; Castejón, Francisco; Gómez-Díez, Manuel; Serrano-Rodríguez, Juan M; Muñoz, Ana

    2014-10-01

    Clinical and laboratory findings were determined in 23 Andalusian horses in southern Spain that were positive for Theileria equi by PCR, including 16 mares at pasture (group A1) and seven stabled stallions (group B1). Five healthy mares at pasture (group A2) and five stabled stallions (group B2), all of which were negative for T. equi in Giemsa stained blood smears and by PCR, were used as controls. The most frequent clinical signs were anorexia, anaemia, depression and icterus (group A1), along with loss of performance or failure to train and depression (group B1). Thrombocytopoenia was evident in 5/7 horses in group B1. Lower serum iron concentrations were observed in both diseased groups compared with their respective control groups. There were no significant differences in APP concentrations between diseased and control groups; all affected horses had APP concentrations within reference limits. Serum haptoglobin, serum amyloid A and plasma fibrinogen concentrations were higher than the reference limits in 5/23, 3/23 and 1/23 diseased horses, respectively. It was concluded that horses with theileriosis exhibited only a mild systemic inflammatory response.

  13. Efficacy of liposomal gentamicin against Rhodococcus equi in a mouse infection model and colocalization with R. equi in equine alveolar macrophages.

    PubMed

    Burton, Alexandra J; Giguère, Steeve; Berghaus, Londa J; Hondalus, Mary K; Arnold, Robert D

    2015-04-17

    Rhodococcus equi, a facultative intracellular pathogen and an important cause of pneumonia in foals, is highly susceptible to killing by gentamicin in vitro. However, gentamicin is not effective in vivo, due to its poor cellular penetration. Encapsulation of drugs in liposomes enhances cellular uptake. The objectives of this study were to compare liposomal gentamicin and free gentamicin with respect to their uptake by equine macrophages and intracellular colocalization with R. equi and to compare the efficacies of liposomal gentamicin, free gentamicin and clarithromycin with rifampin for the reduction of R. equi CFU in a mouse model of infection. After ex vivo exposure, a significantly higher mean (±SD) percentage of equine alveolar macrophages contained liposomal gentamicin (91.9±7.6%) as opposed to free gentamicin (16.8±12.5%). Intracellular colocalization of drug and R. equi, as assessed by confocal microscopy, occurred in a significantly higher proportion of cells exposed to liposomal gentamicin (81.2±17.8%) compared to those exposed to free gentamicin (10.4±8.7%). The number of R. equi CFU in the spleen was significantly lower in mice treated with liposomal gentamicin compared to that of mice treated with free gentamicin or to untreated control mice. Treatment with liposomal gentamicin also resulted in a significantly greater reduction in the number of R. equi CFU in the liver compared to treatment with clarithromycin in combination with rifampin. These results support further investigation of liposomal gentamicin as a new treatment for infections caused by R. equi. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Genomic analyses confirm close relatedness between Rhodococcus defluvii and Rhodococcus equi (Rhodococcus hoagii).

    PubMed

    Sangal, Vartul; Jones, Amanda L; Goodfellow, Michael; Hoskisson, Paul A; Kämpfer, Peter; Sutcliffe, Iain C

    2015-01-01

    Rhodococcus defluvii strain Ca11(T) was isolated from a bioreactor involved in extensive phosphorus removal. We have sequenced the whole genome of this strain, and our comparative genomic and phylogenetic analyses confirm its close relatedness with Rhodococcus equi (Rhodococcus hoagii) strains, which share >80 % of the gene content. The R. equi virulence plasmid is absent though most of the chromosomal R. equi virulence-associated genes are present in R. defluvii Ca11(T). These data suggest that although R. defluvii is an environmental organism, it has the potential to colonize animal hosts.

  15. Activity of clarithromycin or rifampin alone or in combination against experimental Rhodococcus equi infection in mice.

    PubMed

    Burton, Alexandra J; Giguère, Steeve; Berghaus, Londa J; Hondalus, Mary K

    2015-01-01

    Treatment of mice with the combination of clarithromycin with rifampin resulted in a significantly lower number of Rhodococcus equi CFU in the organs of mice than treatment with either drug alone or placebo. There was no significant difference in the number of R. equi CFU between mice treated with clarithromycin monotherapy, rifampin monotherapy, or placebo. The combination of clarithromycin with rifampin conferred a clear advantage over either drug as monotherapy in this model of chronic R. equi infection. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  16. Identification of Rhodococcus equi lipids recognized by host cytotoxic T lymphocytes

    PubMed Central

    Harris, Seth P.; Fujiwara, Nagatoshi; Mealey, Robert H.; Alperin, Debra C.; Naka, Takashi; Goda, Reina

    2010-01-01

    Immune adult horses have CD8+ cytotoxic T lymphocytes (CTLs) that recognize and lyse Rhodococcus equi-infected cells in an equine lymphocyte alloantigen (ELA)-A [classical major histocompatibility complex (MHC) class I]-unrestricted fashion. As protein antigens are MHC class I-restricted, the lack of restriction suggests that the bacterial antigens being recognized by the host are not proteins. The goals of this study were to test the hypothesis that these CTLs recognize unique R. equi cell-wall lipids related to mycobacterial lipids. Initial experiments showed that treatment of soluble R. equi antigen with broadly reactive proteases did not significantly diminish the ability of the antigen to stimulate R. equi-specific CTLs. R. equi-specific CTLs were also shown to lyse target cells (equine macrophages) pulsed with an R. equi lipid extract. Analysis of the R. equi lipid by TLC and MS (MALDI-TOF and ES) indicated that the extracted antigen consisted of three primary fractions: trehalose monomycolate (TMM), trehalose dimycolate (TDM) and cardiolipin (CL). ELA-A-mismatched cells pulsed with purified TMM and CL, but not the TDM fraction, were recognized and lysed by R. equi-specific CTLs. Because of their role in immune clearance and pathogenesis, transcription of the cytokines gamma interferon (IFN-γ) and interleukin-4 (IL-4) was also measured in response to R. equi lipids by using real-time PCR; elevated IFN-γ, but not IL-4, was associated with host clearance of the bacteria. The whole-cell R. equi lipid and all three R. equi lipid fractions resulted in marked increases in IFN-γ transcription, but no increase in IL-4 transcription. Together, these data support the hypothesis that immune recognition of unique lipids in the bacterial cell wall is an important component of the protective immune response to R. equi. The results also identify potential lipid antigens not previously shown to be recognized by CTLs in an important, naturally occurring actinomycete

  17. Evaluations of buparvaquone as a treatment for equine babesiosis (Babesia equi).

    PubMed

    Zaugg, J L; Lane, V M

    1989-05-01

    We evaluated the efficacy of buparvaquone in eliminating Babesia equi of European origin in carrier horses and in experimentally infected splenectomized ponies. When administered at the rate of 2.5 mg/kg of body weight, IM, 4 times at 96-hour intervals, buparvaquone was effective in eliminating B equi carrier infection in 1 horse. Such results could not be repeated at the same dosage or at 3.5 or 5 mg/kg, IM. Buparvaquone given at the rate of 4 to 6 mg/kg IV and/or IM was therapeutically effective in 4 of 5 acute B equi infections in splenectomized ponies. The treated ponies became carriers.

  18. A real-time impedance based method to assess Rhodococcus equi virulence.

    PubMed

    Miranda-CasoLuengo, Aleksandra A; Miranda-CasoLuengo, Raúl; Lieggi, Nora T; Luo, Haixia; Simpson, Jeremy C; Meijer, Wim G

    2013-01-01

    Rhodococcus equi is a facultative intracellular pathogen of macrophages and the causative agent of foal pneumonia. R. equi virulence is usually assessed by analyzing intracellular growth in macrophages by enumeration of bacteria following cell lysis, which is time consuming and does not allow for a high throughput analysis. This paper describes the use of an impedance based real-time method to characterize proliferation of R. equi in macrophages, using virulent and attenuated strains lacking the vapA gene or virulence plasmid. Image analysis suggested that the time-dependent cell response profile (TCRP) is governed by cell size and roundness as well as cytoxicity of infecting R. equi strains. The amplitude and inflection point of the resulting TCRP were dependent on the multiplicity of infection as well as virulence of the infecting strain, thus distinguishing between virulent and attenuated strains.

  19. A Real-Time Impedance Based Method to Assess Rhodococcus equi Virulence

    PubMed Central

    Lieggi, Nora T.; Luo, Haixia; Simpson, Jeremy C.; Meijer, Wim G.

    2013-01-01

    Rhodococcus equi is a facultative intracellular pathogen of macrophages and the causative agent of foal pneumonia. R. equi virulence is usually assessed by analyzing intracellular growth in macrophages by enumeration of bacteria following cell lysis, which is time consuming and does not allow for a high throughput analysis. This paper describes the use of an impedance based real-time method to characterize proliferation of R. equi in macrophages, using virulent and attenuated strains lacking the vapA gene or virulence plasmid. Image analysis suggested that the time-dependent cell response profile (TCRP) is governed by cell size and roundness as well as cytoxicity of infecting R. equi strains. The amplitude and inflection point of the resulting TCRP were dependent on the multiplicity of infection as well as virulence of the infecting strain, thus distinguishing between virulent and attenuated strains. PMID:23555995

  20. Immune Reconstitution Syndrome secondary to Rhodococcus equi infection in a patient with HIV and Burkitt's lymphoma.

    PubMed

    Darraj, Majid; Fainstein, Rachel; Kasper, Ken; Keynan, Yoav

    Immune Reconstitution Syndrome (IRIS) has been associated with a variety of infections in patients with human immunodeficiency virus (HIV). However, we are reporting the first case of IRIS secondary to Rhodococcus equi (R. equi) in a patient with HIV. We report the case of a 48-year-old male found to have HIV infection in the setting of Burkitt's lymphoma. While on anti-retroviral therapy and chemotherapy, he had developed IRIS secondary to R. equi that manifested as a cavitating pneumonia. This report outlines the successful management of the R. equi infection with the use of a combination of antibiotics, radiographic follow up and suppressive antibiotic while on chemotherapy. Copyright © 2016 King Saud Bin Abdulaziz University for Health Sciences. Published by Elsevier Ltd. All rights reserved.

  1. The correct name of the taxon that contains the type strain of Rhodococcus equi.

    PubMed

    Tindall, B J

    2014-01-01

    Based on a nomenclatural point of view, the name Rhodococcus equi is associated, as required by the Bacteriological Code, with a defined position, rank and circumscription. A search of the literature indicates that the name Rhodococcus equi (Magnusson 1923) Goodfellow and Alderson 1977 has also been shown to be a synonym of Corynebacterium equi Magnusson 1923, Corynebacterium hoagii (Morse 1912) Eberson 1918 and Nocardia restricta (Turfitt 1944) McClung 1974. Application of the rules of the Bacteriological Code together with the currently inferred taxonomic concept associated with the species bearing the name Rhodococcus equi indicates that this is not the correct name of this taxon and the use of that name in the context of a circumscription that includes the type strain of the species Corynebacterium hoagii is contrary to the Rules of the Code.

  2. Implementing the EQUiPPED Medication Management Program at 5 VA Emergency Departments.

    PubMed

    Vandenberg, Ann E; Stevens, Melissa; Echt, Katharina V; Hastings, S Nicole; Powers, James; Markland, Alayne; Hwang, Ula; Hung, William; Belbis, Stephanie; Vaughan, Camille P

    2016-04-01

    The Enhancing Quality of Prescribing Practices for Older Veterans Discharged From the Emergency Department (EQUiPPED) program aimed to reduce potentially inappropriate medication prescribing to older adults at 5 VAMCs.

  3. Corynebacterium equi: in vitro susceptibility to twenty-six antimicrobial agents.

    PubMed Central

    Woolcock, J B; Mutimer, M D

    1980-01-01

    The minimal concentrations of 26 antimicrobial agents required to inhibit growth of 100 isolates of Corynebacterium equi in vitro have been determined. The most active agents were penicillin G, doxycycline, erythromycin, lincomycin, and the aminoglycosides. PMID:7235683

  4. Novel transferable erm(46) determinant responsible for emerging macrolide resistance in Rhodococcus equi.

    PubMed

    Anastasi, Elisa; Giguère, Steeve; Berghaus, Londa J; Hondalus, Mary K; Willingham-Lane, Jennifer M; MacArthur, Iain; Cohen, Noah D; Roberts, Marilyn C; Vazquez-Boland, Jose A

    2015-12-01

    The objective of this study was to identify the molecular mechanism of macrolide resistance in the actinomycete Rhodococcus equi, a major equine pathogen and zoonotic agent causing opportunistic infections in people. Macrolide-resistant (n = 62) and macrolide-susceptible (n = 62) clinical isolates of R. equi from foals in the USA were studied. WGS of 18 macrolide-resistant and 6 macrolide-susceptible R. equi was performed. Representative sequences of all known macrolide resistance genes identified to date were used to search the genome assemblies for putative homologues. PCR was used to screen for the presence of the identified resistance determinant in the rest of the isolates. Mating experiments were performed to verify mobility of the gene. A novel erm gene, erm(46), was identified in all sequenced resistant isolates, but not in susceptible isolates. There was complete association between macrolide resistance and the presence of erm(46) as detected by PCR screening of all 124 clinical isolates of R. equi. Expression of erm(46) in a macrolide-susceptible strain of R. equi induced high-level resistance to macrolides, lincosamides and streptogramins B, but not to other classes of antimicrobial agents. Transfer of erm(46) to macrolide-susceptible R. equi was confirmed. The transfer frequency ranged from 3 × 10(-3) to 1 × 10(-2). This is the first molecular characterization of resistance to macrolides, lincosamides and streptogramins B in R. equi. Resistance was due to the presence of a novel erm(46) gene mobilizable likely by conjugation, which has spread among equine isolates of R. equi in the USA. © The Author 2015. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  5. Effects of opsonization of Rhodococcus equi on bacterial viability and phagocyte activation.

    PubMed

    Dawson, Dominic R; Nydam, Daryl V; Price, Christopher T; Graham, James E; Cynamon, Michael H; Divers, Thomas J; Felippe, Maria Julia B

    2011-11-01

    To investigate the effect of opsonization of Rhodococcus equi with R. equi-specific antibodies in plasma on bacterial viability and phagocyte activation in a cell culture model of infection. Neutrophils and monocyte-derived macrophages from 6 healthy 1-week-old foals and 1 adult horse. Foal and adult horse phagocytes were incubated with either opsonized or nonopsonized bacteria. Opsonization was achieved by use of plasma containing high or low concentrations of R. equi-specific antibodies. Phagocyte oxidative burst activity was measured by use of flow cytometry, and macrophage tumor necrosis factor (TNF)-α production was measured via an ELISA. Extracellular and intracellular bacterial viability was measured with a novel R. equi-luciferase construct that used a luminometer. Opsonized bacteria increased oxidative burst activity in adult horse phagocytes, and neutrophil activity was dependent on the concentration of specific antibody. Secretion of TNF-α was higher in macrophages infected with opsonized bacteria. Opsonization had no significant effect on bacterial viability in macrophages; however, extracellular bacterial viability was decreased in broth containing plasma with R. equi-specific antibodies, compared with viability in broth alone. The use of plasma enriched with specific antibodies for the opsonization of R. equi increased the activation of phagocytes and decreased bacterial viability in the extracellular space. Although opsonized R. equi increased TNF-α secretion and oxidative burst in macrophages, additional factors may be necessary for effective intracellular bacterial killing. These data have suggested a possible role of plasma antibody in protection of foals from R. equi pneumonia.

  6. The effect of bacterial dose and foal age at challenge on Rhodococcus equi infection.

    PubMed

    Sanz, M; Loynachan, A; Sun, L; Oliveira, A; Breheny, P; Horohov, D W

    2013-12-27

    While Rhodococcus equi remains the most common cause of subacute or chronic granulomatous bronchopneumonia in foals, development of a relevant model to study R. equi infection has proven difficult. The objective of this study was to identify a challenge dose of R. equi that resulted in slow progressive disease, spontaneous regression of lung lesions and age-dependent susceptibility. Foals less than one-week of age were challenged intratracheally using either 10(6), 10(5), 10(4), 10(3) or 10(2) cfu of R. equi. Two doses (10(3) cfu and 10(5) cfu) were used to challenge 2 and 3-week-old, and 3 and 6-week-old foals, respectively. Physical examination, thoracic ultrasound and blood work were performed. Foals were euthanized at the end of the study or when clinical signs of pneumonia developed. All foals were necropsied and their lung lesions scored. Foals challenged with low concentrations of R. equi developed slow progressive pneumonia and approximately 50% of the foals recovered spontaneously. Likewise, macroscopic (>1cm diameter) pyogranulomatous lesions were only observed when low doses of R. equi were used. Clinical pneumonia was not seen after low dose challenge in the 3-week-old foals or in the 6-week-old foals. This study demonstrates that the use of low doses of R. equi to challenge neonatal foals provides an improved model for studying this disease. Furthermore, susceptibility to R. equi infection was shown to diminish early in the foal's life, as has been reported in the field. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Relationship of mixed bacterial infection to prognosis in foals with pneumonia caused by Rhodococcus equi.

    PubMed

    Giguère, S; Jordan, L M I; Glass, K; Cohen, N D

    2012-01-01

    Isolation of multiple bacterial species is common in foals with Rhodococcus equi pneumonia. There is no association between isolation of other microorganisms and outcome. 155 foals with pneumonia caused by R. equi. Case records of foals diagnosed with R. equi pneumonia based on culture of the respiratory tract were reviewed at 2 referral hospitals (University of Florida [UF] and Texas A&M University [TAMU]). R. equi was cultured from a tracheobronchial aspirate (TBA) in 115 foals and from lung tissue in 38 foals. Survival was significantly higher at UF (71%; 70/99) than at TAMU (50%; 28/56). R. equi was significantly more likely to grow in pure cultures from samples obtained from foals at UF (55%; 54/99) than from foals at TAMU (23%; 13/56). Microorganisms cultured with R. equi included Gram-positive bacteria in 40, Gram-negative bacteria in 41, and fungi in 23 foals. The most common bacteria isolated were beta-hemolytic streptococci (n = 26) and Escherichia coli (n = 18). Mixed infections were significantly more likely to be encountered in TBA than in lung tissue. Only foals from which R. equi was cultured from a TBA were included in the analysis for association between mixed infection and outcome. After adjusting for the effect of hospital using multivariate logistic regression, mixed culture, mixed bacterial culture, Gram-positive bacteria, beta-hemolytic streptococci, Gram-negative bacteria, enteric Gram-negative bacteria, nonenteric Gram-negative bacteria, and fungi were not significantly associated with outcome. Isolation of multiple bacteria or fungi from a TBA along with R. equi does not negatively impact prognosis. Copyright © 2012 by the American College of Veterinary Internal Medicine.

  8. Use of Liposomal Gentamicin for Treatment of 5 Foals with Experimentally Induced Rhodococcus equi Pneumonia.

    PubMed

    Cohen, N D; Giguère, S; Burton, A J; Rocha, J N; Berghaus, L J; Brake, C N; Bordin, A I; Coleman, M C

    2016-01-01

    Adverse effects of, and bacterial resistance to, macrolides used to treat Rhodococcus equi infections have prompted search for clinically effective alternative antimicrobials. Liposomal gentamicin (LG) is effective against R. equi in vitro and decreases tissue concentrations of R. equi in experimentally infected mice. Effectiveness of LG treatment of foals with R. equi pneumonia, however, has not been described. Liposomal gentamicin is safe and effective for treating foals with R. equi pneumonia. Ten foals with experimentally induced R. equi pneumonia. Pilot treatment trial. Foals with pneumonia induced by intrabronchial instillation of R. equi were randomly allocated to receive either clarithromycin combined with rifampin (CLR + RIF) PO or LG IV, and followed by daily physical examinations and weekly thoracic ultrasonography and serum creatinine concentration determinations until the resolution of clinical signs. Treatment success was defined as the resolution of clinical signs and ultrasonographically identified pulmonary abscesses. All 10 foals were successfully treated. Two of 5 foals treated with LG developed azotemia within 1 week; LG was discontinued and treatment switched to CLR + RIF for these foals. None of the CLR + RIF treated foals developed azotemia. Liposomal gentamicin IV can be effective for treatment of R. equi pneumonia, but nephrotoxicity indicates that an alternative dosing interval or route (such as nebulization) will be needed before LG is adequately safe for clinical use. Larger comparative trials will be needed to evaluate the relative efficacy of a safer LG dosage regimen. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  9. In vitro synergy, pharmacodynamics, and postantibiotic effect of 11 antimicrobial agents against Rhodococcus equi.

    PubMed

    Giguère, Steeve; Lee, Elise A; Guldbech, Kristen M; Berghaus, Londa J

    2012-11-09

    There are no studies investigating interactions between clarithromycin or azithromycin and rifampin or other commonly used antimicrobial agents against virulent isolates of Rhodococcus equi. In addition, there is no published data on the postantibiotic effects (PAEs) and pharmacodynamics properties of antimicrobial agents against R. equi. The objectives were to assess in vitro interactions, pharmacodynamics, and PAEs of 11 antimicrobial agents belonging to various antimicrobial classes against R. equi. Antimicrobial agents investigated (erythromycin, clarithromycin, azithromycin, rifampin, amikacin, gentamicin, enrofloxacin, vancomycin, imipenem, ceftiofur, and doxycycline) were selected based on in vitro activity against large numbers of isolates of R. equi and frequency of use in foals or humans infected with R. equi. Three virulent strains of R. equi were evaluated by time-kill curves and checkerboard assays, and the postantibiotic effect was measured at 5×MIC. Only amikacin, gentamicin, enrofloxacin, and vancomycin were bactericidal against R. equi. Combinations including a macrolide (erythromycin, clarithromycin, azithromycin) and either rifampin or doxycycline, and the combination doxycycline-rifampin were synergistic. Combinations containing amikacin and erythromycin, clarithromycin, azithromycin, or rifampin and the combination gentamicin-rifampin were antagonistic. The PAEs of rifampin, erythromycin, clarithromycin, vancomycin, and doxycycline were relatively long with median values ranging between 4.5 and 6.5h. Azithromycin, gentamicin, and imipenem had intermediate PAEs ranging between 3.3 and 3.5h. Amikacin, enrofloxacin, and ceftiofur had shorter PAEs ranging between 1.3 and 2.1h. Gentamicin, amikacin, enrofloxacin, and doxycycline exhibited concentration-dependent activity whereas erythromycin, clarithromycin, azithromycin, rifampin, ceftiofur, imipenem, and vancomycin exhibited time-dependent activity against R. equi. Copyright © 2012 Elsevier B

  10. Demographic and environmental risk factors for infection by Theileria equi in 590 horses in Israel.

    PubMed

    Steinman, Amir; Zimmerman, Tal; Klement, Eyal; Lensky, Itamar M; Berlin, Dalia; Gottlieb, Yuval; Baneth, Gad

    2012-07-06

    The prevalence of Theileria equi infection as well as the environmental and demographic risk factors for infection was studied in 590 healthy horses from 46 farms in Israel. The prevalence of T. equi DNA was assessed using a polymerase chain reaction for a segment of the Theileria 18S rRNA gene. The overall prevalence was 26.4% (156/590). There was a significant geographical variation in the prevalence of T. equi infection, ranging from 9.3% (25/270) in the central lowlands to 81.7% (49/60) in the Golan Heights. The prevalence of T. equi infection was found to be significantly associated with management types with more horses with access to pasture being positive. Breed was identified as a risk factor for T. equi infection in a univariate analysis with relatively high infection rates in the Quarter horse and local breeds (41.1% and 36.3% respectively), while ponies and Arabian horses had a relatively low prevalence (10% and 9.1%, respectively). However, since a correlation between geographic location and breed was found, it is difficult to draw definite conclusions regarding this risk factor. Age and gender were not found as risk factors for T. equi infection in this study. The environmental variables that were significantly associated with positivity were relative humidity and minimum land surface temperature at day which both showed negative correlation with T. equi prevalence. In conclusion, Israel was found to be enzootic for T. equi infection, as indicated by the high sub-clinical infection rate, which differed between geographical areas.

  11. Molecular epidemiology of Rhodococcus equi in slaughtered swine, cattle and horses in Poland.

    PubMed

    Witkowski, Lucjan; Rzewuska, Magdalena; Takai, Shinji; Kizerwetter-Świda, Magdalena; Kita, Jerzy

    2016-05-27

    Rhodococcus equi is an emerging zoonotic presumably foodborne pathogen. Since the data on the worldwide prevalence of R. equi in meat animals are scarce, the present study aimed to investigate the molecular epidemiology of R. equi in swine, cattle and horse carcasses intended for human consumption in Poland. Totally 1028 lymph node samples were examined. R. equi was isolated from 26.6 % (105/395) swine and 1.3 % (3/234) bovine healthy submaxillary lymph nodes. In horses, R. equi was isolated only from 0.5 % (1/198) samples of middle tracheo-branchiales lymph node while no lymphocentrum retropharyngeum sample was positive (0/198). The purulent lesions were observed only in 0.8 % swine submaxillary lymph nodes samples (3/398) and in two of them R. equi was detected. All bovine and most of swine isolates (98.1 %) were vapB-positive. 87.9 % of swine isolates carried 95-kb type 5 plasmid, 3.7 % type 1 and plasmid types: 4, 7, 10, 11, 21, 31 were carried by a single isolate (0.9 %). All bovine isolates carried VAPB type 26. Single horse isolate was vapA-positive and carried plasmid VAPA 85-kb type I. The prevalence of vapB-positive R. equi in investigated healthy swine intended for human consumption was very high. Not only swine, but also even apparently healthy cattle or horse carcasses should be considered as a potential source of R. equi for humans, especially in countries where undercooked or raw beef or horsemeat is traditionally consumed.

  12. Unusual Outbreak of Clinical Mastitis in Dairy Sheep Caused by Streptococcus equi subsp. zooepidemicus

    PubMed Central

    Las Heras, Alfonso; Vela, Ana I.; Fernández, Elena; Legaz, Emilio; Domínguez, Lucas; Fernández-Garayzábal, Jose F.

    2002-01-01

    This work describes an outbreak of clinical mastitis affecting 13 of 58 lactating ewes due to Streptococcus equi subsp. zooepidemicus. S. equi subsp. zooepidemicus was isolated in pure culture from all milk samples. All the clinical isolates had identical biochemical profiles and antimicrobial susceptibility patterns and also exhibited indistinguishable macrorestriction patterns by pulsed-field gel electrophoresis, indicating that all cases of mastitis were produced by a single strain. PMID:11880454

  13. Unusual outbreak of clinical mastitis in dairy sheep caused by Streptococcus equi subsp. zooepidemicus.

    PubMed

    Las Heras, Alfonso; Vela, Ana I; Fernández, Elena; Legaz, Emilio; Domínguez, Lucas; Fernández-Garayzábal, Jose F

    2002-03-01

    This work describes an outbreak of clinical mastitis affecting 13 of 58 lactating ewes due to Streptococcus equi subsp. zooepidemicus. S. equi subsp. zooepidemicus was isolated in pure culture from all milk samples. All the clinical isolates had identical biochemical profiles and antimicrobial susceptibility patterns and also exhibited indistinguishable macrorestriction patterns by pulsed-field gel electrophoresis, indicating that all cases of mastitis were produced by a single strain.

  14. Crossiella equi sp. nov., isolated from equine placentas.

    PubMed

    Donahue, J M; Williams, N M; Sells, S F; Labeda, D P

    2002-11-01

    Over the course of the past decade, actinomycetes have been isolated from the placentas of horses diagnosed with nocardioform placentitis. The incidence of this infection has generally been low, with typically no more than 30 animals affected in most years, but the incidence increased through 1999, with placentas from 144 mares found to be infected. Approximately half of the cases result in loss of the foal. A typical actinomycete with branching mycelium was isolated from placental lesions, and a comparison of the sequence of the 16S rDNA gene against the public databases indicated a relationship to members of the suborder Pseudonocardineae. Phylogenetic analysis of representative isolates revealed a close relationship to Crossiella cryophila, and subsequent polyphasic comparisons determined that these isolates represent a novel species of Crossiella, for which the name Crossiella equi sp. nov. is proposed, with strain LDDC 22291-98(T) (= NRRL B-24104(T) = DSM 44580(T)) as the type strain.

  15. Purulent pericarditis and pneumonia caused by Streptococcus equi subsp. zooepidemicus.

    PubMed

    Held, Jürgen; Schmitz, Roland; van der Linden, Mark; Nührenberg, Thomas; Häcker, Georg; Neumann, Franz-Josef

    2014-02-01

    Purulent pericarditis is a life-threatening disease that usually manifests following bacteraemia or through spreading from an intrathoracic focus. Only a few cases of this disease have been reported with Lancefield group C streptococci as aetiological agents, and the primary focus in these infections remains unknown. We report a case of purulent pericarditis with septic and cardiogenic shock, caused by Streptococcus equi subsp. zooepidemicus (group C) in a 51-year-old patient. The pathogen was possibly contracted through contact with horses. Most probably, it caused initially pneumonia before spreading to the pericardium, either directly or via the bloodstream. A combined therapeutic approach, consisting of antibiotic therapy and repeated pericardial drainage, was necessary to ensure a clinical cure. After discharge, long-term follow-up for development of constrictive pericarditis is considered mandatory.

  16. Nitric oxide-mediated intracellular growth restriction of pathogenic Rhodococcus equi can be prevented by iron.

    PubMed

    von Bargen, Kristine; Wohlmann, Jens; Taylor, Gregory Alan; Utermöhlen, Olaf; Haas, Albert

    2011-05-01

    Rhodococcus equi is an intracellular pathogen which causes pneumonia in young horses and in immunocompromised humans. R. equi arrests phagosome maturation in macrophages at a prephagolysosome stage and grows inside a privileged compartment. Here, we show that, in murine macrophages activated with gamma interferon and lipopolysaccharide, R. equi does not multiply but stays viable for at least 24 h. Whereas infection control of other intracellular pathogens by activated macrophages is executed by enhanced phagosome acidification or phagolysosome formation, by autophagy or by the interferon-inducible GTPase Irgm1, none of these mechanisms seems to control R. equi infection. Growth control by macrophage activation is fully mimicked by treatment of resting macrophages with nitric oxide donors, and inhibition of bacterial multiplication by either activation or nitric oxide donors is annihilated by cotreatment of infected macrophages with ferrous sulfate. Transcriptional analysis of the R. equi iron-regulated gene iupT demonstrates that intracellular R. equi encounters iron stress in activated, but not in resting, macrophages and that this stress is relieved by extracellular addition of ferrous sulfate. Our results suggest that nitric oxide is central to the restriction of bacterial access to iron in activated macrophages.

  17. Induction of proinflammatory cytokines in human lung epithelial cells during Rhodococcus equi infection.

    PubMed

    Remuzgo-Martínez, Sara; Pilares-Ortega, Lilian; Alvarez-Rodríguez, Lorena; Aranzamendi-Zaldunbide, Maitane; Padilla, Daniel; Icardo, Jose Manuel; Ramos-Vivas, Jose

    2013-08-01

    Rhodococcus equi is an opportunistic human pathogen associated with immunosuppressed people. While the interaction of R. equi with macrophages has been comprehensively studied, little is known about its interactions with non-phagocytic cells. Here, we characterized the entry process of this bacterium into human lung epithelial cells. The invasion is inhibited by nocodazole and wortmannin, suggesting that the phosphatidylinositol 3-kinase pathway and microtubule cytoskeleton are important for invasion. Pre-incubation of R. equi with a rabbit anti-R. equi polyclonal antiserum resulted in a dramatic reduction in invasion. Also, the invasion process as studied by immunofluorescence and scanning electron microscopy indicates that R. equi make initial contact with the microvilli of the A549 cells, and at the structural level, the entry process was observed to occur via a zipper-like mechanism. Infected lung epithelial cells upregulate the expression of cytokines IL-8 and IL-6 upon infection. The production of these pro-inflammatory cytokines was significantly enhanced in culture supernatants from cells infected with non-mucoid plasmid-less strains when compared with cells infected with mucoid strains. These results demonstrate that human airway epithelial cells produce pro-inflammatory mediators against R. equi isolates.

  18. Nitric Oxide-Mediated Intracellular Growth Restriction of Pathogenic Rhodococcus equi Can Be Prevented by Iron▿

    PubMed Central

    von Bargen, Kristine; Wohlmann, Jens; Taylor, Gregory Alan; Utermöhlen, Olaf; Haas, Albert

    2011-01-01

    Rhodococcus equi is an intracellular pathogen which causes pneumonia in young horses and in immunocompromised humans. R. equi arrests phagosome maturation in macrophages at a prephagolysosome stage and grows inside a privileged compartment. Here, we show that, in murine macrophages activated with gamma interferon and lipopolysaccharide, R. equi does not multiply but stays viable for at least 24 h. Whereas infection control of other intracellular pathogens by activated macrophages is executed by enhanced phagosome acidification or phagolysosome formation, by autophagy or by the interferon-inducible GTPase Irgm1, none of these mechanisms seems to control R. equi infection. Growth control by macrophage activation is fully mimicked by treatment of resting macrophages with nitric oxide donors, and inhibition of bacterial multiplication by either activation or nitric oxide donors is annihilated by cotreatment of infected macrophages with ferrous sulfate. Transcriptional analysis of the R. equi iron-regulated gene iupT demonstrates that intracellular R. equi encounters iron stress in activated, but not in resting, macrophages and that this stress is relieved by extracellular addition of ferrous sulfate. Our results suggest that nitric oxide is central to the restriction of bacterial access to iron in activated macrophages. PMID:21383050

  19. Characterization of Pneumonia Due to Streptococcus equi subsp. zooepidemicus in Dogs▿

    PubMed Central

    Priestnall, Simon L.; Erles, Kerstin; Brooks, Harriet W.; Cardwell, Jacqueline M.; Waller, Andrew S.; Paillot, Romain; Robinson, Carl; Darby, Alistair C.; Holden, Matthew T. G.; Schöniger, Sandra

    2010-01-01

    Streptococcus equi subsp. zooepidemicus has been linked to cases of acute fatal pneumonia in dogs in several countries. Outbreaks can occur in kenneled dog populations and result in significant levels of morbidity and mortality. This highly contagious disease is characterized by the sudden onset of clinical signs, including pyrexia, dyspnea, and hemorrhagic nasal discharge. The pathogenesis of S. equi subsp. zooepidemicus infection in dogs is poorly understood. This study systematically characterized the histopathological changes in the lungs of 39 dogs from a large rehoming shelter in London, United Kingdom; the dogs were infected with S. equi subsp. zooepidemicus. An objective scoring system demonstrated that S. equi subsp. zooepidemicus caused pneumonia in 26/39 (66.7%) dogs, and most of these dogs (17/26 [65.4%]) were classified as severe fibrino-suppurative, necrotizing, and hemorrhagic. Three recently described superantigen genes (szeF, szeN, and szeP) were detected by PCR in 17/47 (36.2%) of the S. equi subsp. zooepidemicus isolates; however, there was no association between the presence of these genes and the histopathological score. The lungs of S. equi subsp. zooepidemicus-infected dogs with severe respiratory signs and lung pathology did however have significantly higher mRNA levels of the proinflammatory cytokines tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), and interleukin 8 (IL-8) than in uninfected controls, suggesting a role for an exuberant host immune response in the pathogenesis of this disease. PMID:20861329

  20. EquiFACS: The Equine Facial Action Coding System

    PubMed Central

    Wathan, Jen; Burrows, Anne M.; Waller, Bridget M.; McComb, Karen

    2015-01-01

    Although previous studies of horses have investigated their facial expressions in specific contexts, e.g. pain, until now there has been no methodology available that documents all the possible facial movements of the horse and provides a way to record all potential facial configurations. This is essential for an objective description of horse facial expressions across a range of contexts that reflect different emotional states. Facial Action Coding Systems (FACS) provide a systematic methodology of identifying and coding facial expressions on the basis of underlying facial musculature and muscle movement. FACS are anatomically based and document all possible facial movements rather than a configuration of movements associated with a particular situation. Consequently, FACS can be applied as a tool for a wide range of research questions. We developed FACS for the domestic horse (Equus caballus) through anatomical investigation of the underlying musculature and subsequent analysis of naturally occurring behaviour captured on high quality video. Discrete facial movements were identified and described in terms of the underlying muscle contractions, in correspondence with previous FACS systems. The reliability of others to be able to learn this system (EquiFACS) and consistently code behavioural sequences was high—and this included people with no previous experience of horses. A wide range of facial movements were identified, including many that are also seen in primates and other domestic animals (dogs and cats). EquiFACS provides a method that can now be used to document the facial movements associated with different social contexts and thus to address questions relevant to understanding social cognition and comparative psychology, as well as informing current veterinary and animal welfare practices. PMID:26244573

  1. Use of Serial Quantitative PCR of the vapA Gene of Rhodococcus equi in Feces for Early Detection of R. equi Pneumonia in Foals.

    PubMed

    Madrigal, R G; Shaw, S D; Witkowski, L A; Sisson, B E; Blodgett, G P; Chaffin, M K; Cohen, N D

    2016-01-01

    Current screening tests for Rhodococcus equi pneumonia in foals lack adequate accuracy for clinical use. Real-time, quantitative PCR (qPCR) for virulent R. equi in feces has not been systematically evaluated as a screening test. The objective of this study was to evaluate the accuracy of qPCR for vapA in serially collected fecal samples as a screening test for R. equi pneumonia in foals. One hundred and twenty-five foals born in 2011 at a ranch in Texas. Fecal samples were collected concurrently with thoracic ultrasonography (TUS) screening examinations at ages 3, 5, and 7 weeks. Affected (pneumonic) foals (n = 25) were matched by age and date-of-birth to unaffected (n = 25) and subclinical (ie, having thoracic TUS lesions but no clinical signs of pneumonia) foals (n = 75). DNA was extracted from feces using commercial kits and concentration of virulent R. equi in feces was determined by qPCR. Subsequently affected foals had significantly greater concentrations of vapA in feces than foals that did not develop pneumonia (unaffected and subclinical foals) at 5 and 7 weeks of age. Accuracy of fecal qPCR, however, was poor as a screening test to differentiate foals that would develop clinical signs of pneumonia from those that would remain free of clinical signs (including foals with subclinical pulmonary lesions attributed to R. equi) using receiver operating characteristic (ROC) methods. In the population studied, serial qPCR on feces lacked adequate accuracy as a screening test for clinical R. equi foal pneumonia. Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  2. Bronchopneumonia in wild boar (Sus scrofa) caused by Rhodococcus equi carrying the VapB type 8 plasmid.

    PubMed

    de Vargas, Agueda Castagna; Monego, Fernanda; Gressler, Letícia Trevisan; de Avila Botton, Sônia; Lazzari, Andrea Maria; da Costa, Mateus Matiuzzi; Ecco, Roselene; Ribeiro, Márcio Garcia; Lara, Gustavo Henrique Batista; Takai, Shinji

    2013-03-25

    Rhodococcus equi is associated with pyogranulomatous infections, especially in foals, and this bacterium has also emerged as a pathogen for humans, particularly immunocompromised patients. R. equi infections in pigs, wild boar (Sus scrofa) and humans are mainly due to strains carrying the intermediate virulence (VapB) plasmid. In Brazil, R. equi carrying the VapB type 8 plasmid is the most common type recovered from humans co-infected with the human immunodeficiency virus (HIV). R. equi infection in pigs and wild boar is restricted predominantly to the lymphatic system, without any reports of pulmonary manifestations. This report describes the microbiological and histopathological findings, and molecular characterization of R. equi in two bronchopneumonia cases in wild boar using PCR and plasmid profile analysis by digestion with restriction endonucleases. The histological findings were suggestive of pyogranulomatous infection, and the plasmid profile of both R. equi isolates enabled the characterization of the strains as VapB type 8. This is the first report of bronchopneumonia in wild boar due to R. equi. The detection of the VapB type 8 plasmid in R. equi isolates emphasize that wild boar may be a potential source of pathogenic R. equi strains for humans.

  3. Identification and characterization of a Streptococcus equi ssp. zooepidemicus immunogenic GroEL protein involved in biofilm formation.

    PubMed

    Yi, Li; Wang, Yang; Ma, Zhe; Lin, Hui-Xing; Xu, Bin; Grenier, Daniel; Fan, Hong-Jie; Lu, Cheng-Ping

    2016-04-18

    Streptococcus equi ssp. zooepidemicus (S. equi spp. zooepidemicus) is an opportunistic pathogen that causes major economic losses in the swine industry in China and is also a threat for human health. Biofilm formation by this bacterium has been previously reported. In this study, we used an immunoproteomic approach to search for immunogenic proteins expressed by biofilm-grown S. equi spp. zooepidemicus. Seventeen immunoreactive proteins were found, of which nine common immunoreactive proteins were identified in planktonic and biofilm-grown bacteria. The immunogenicity and protective efficacy of the S. equi spp. zooepidemicus immunoreactive GroEL chaperone protein was further investigated in mice. The protein was expressed in vivo and elicited high antibody titers following S. equi spp. zooepidemicus infections of mice. An animal challenge experiment with S. equi spp. zooepidemicus showed that 75% of mice immunized with the GroEL protein were protected. Using in vitro biofilm inhibition assays, evidence was obtained that the chaperonin GroEL may represent a promising target for the prevention and treatment of persistent S. equi spp. zooepidemicus biofilm infections. In summary, our results suggest that the recombinant GroEL protein, which is involved in biofilm formation, may efficiently stimulate an immune response, which protects against S. equi spp. zooepidemicus infections. It may therefore be a candidate of interest to be included in vaccines against S. equi spp. zooepidemicus infections.

  4. Genomic Evidence for the Evolution of Streptococcus equi: Host Restriction, Increased Virulence, and Genetic Exchange with Human Pathogens

    PubMed Central

    Paillot, Romain; Steward, Karen F.; Webb, Katy; Ainslie, Fern; Jourdan, Thibaud; Bason, Nathalie C.; Holroyd, Nancy E.; Mungall, Karen; Quail, Michael A.; Sanders, Mandy; Simmonds, Mark; Willey, David; Brooks, Karen; Aanensen, David M.; Spratt, Brian G.; Jolley, Keith A.; Maiden, Martin C. J.; Kehoe, Michael; Chanter, Neil; Bentley, Stephen D.; Robinson, Carl; Maskell, Duncan J.; Parkhill, Julian; Waller, Andrew S.

    2009-01-01

    The continued evolution of bacterial pathogens has major implications for both human and animal disease, but the exchange of genetic material between host-restricted pathogens is rarely considered. Streptococcus equi subspecies equi (S. equi) is a host-restricted pathogen of horses that has evolved from the zoonotic pathogen Streptococcus equi subspecies zooepidemicus (S. zooepidemicus). These pathogens share approximately 80% genome sequence identity with the important human pathogen Streptococcus pyogenes. We sequenced and compared the genomes of S. equi 4047 and S. zooepidemicus H70 and screened S. equi and S. zooepidemicus strains from around the world to uncover evidence of the genetic events that have shaped the evolution of the S. equi genome and led to its emergence as a host-restricted pathogen. Our analysis provides evidence of functional loss due to mutation and deletion, coupled with pathogenic specialization through the acquisition of bacteriophage encoding a phospholipase A2 toxin, and four superantigens, and an integrative conjugative element carrying a novel iron acquisition system with similarity to the high pathogenicity island of Yersinia pestis. We also highlight that S. equi, S. zooepidemicus, and S. pyogenes share a common phage pool that enhances cross-species pathogen evolution. We conclude that the complex interplay of functional loss, pathogenic specialization, and genetic exchange between S. equi, S. zooepidemicus, and S. pyogenes continues to influence the evolution of these important streptococci. PMID:19325880

  5. Air sampling in the breathing zone of neonatal foals for prediction of subclinical Rhodococcus equi infection.

    PubMed

    Chicken, C; Muscatello, G; Freestone, J; Anderson, G A; Browning, G F; Gilkerson, J R

    2012-03-01

    Disease caused by Rhodococcus equi is a significant burden to the horse breeding industry worldwide. Early detection of rhodococcal pneumonia, albeit important to minimise treatment costs, is difficult because of the insidious nature of the disease and the lack of definitive diagnostic tests. To investigate air sampling from the breathing zone of neonatal foals as a predictor of subsequent rhodococcal pneumonia. Air samples were collected from the breathing zone of 53 neonatal foals (age ≤10 days) and again at the time of routine ultrasonographic screening for R. equi pneumonia (age 1-2 months). Pneumonia was diagnosed ultrasonographically in 23% of foals. Virulent R. equi was detected in air from the breathing zone of 19% of neonatal foals and 45% of foals at age 1-2 months. There was no association between virulent R. equi in the breathing zone of foals and the subsequent ultrasonographic diagnosis of rhodococcal pneumonia. The median concentration of virulent R. equi in the breathing zone of both neonates (0 [range 0-4] colony-forming units [cfu]/250 l) and older foals (0 [range 0-3] cfu/250 l) was not significantly different from that in background air samples (0 [range 0-6] cfu/250 l). There was no difference in the concentration of virulent R. equi in the breathing zone of older foals that were diagnosed with rhodococcal pneumonia or clinically normal foals. Detection of virulent R. equi in air from the breathing zone was not a positive predictor of rhodococcal pneumonia in foals up to age ≤2 months. Selective culture of air samples from the breathing zone of young foals is not better at diagnosing rhodococcal pneumonia than early ultrasonographic screening. However, culture of air samples from the breathing zone of older foals remains a useful herd-based epidemiological tool. © 2012 EVJ Ltd.

  6. Oxidative stress and DNA damage in horses naturally infected with Theileria equi.

    PubMed

    Radakovic, M; Davitkov, D; Borozan, S; Stojanovic, S; Stevanovic, J; Krstic, V; Stanimirovic, Z

    2016-11-01

    The aim of this study was to determine the concentrations of oxidative stress parameters and DNA damage in horses infected by Theileria equi. Initial screening of 110 horses with duplex PCR enabled the selection of 30 infected horses with T. equi and 30 free of infection (control). Specimens from the 60 horses were further analysed by determining the following oxidative stress parameters: extent of haemolysis (EH), plasma free haemoglobin (PHb), catalase (CAT), Cu,Zn superoxide dismutase (SOD1), paraoxonase (PON1), nitrite (NO2(-)), total nitrate and nitrite (NOx), malondialdehyde (MDA) and free thiol groups (-SH). In addition, relative distribution of lactate dehydrogenase (LDH1-LDH5) activity and the DNA-damaging effects of T. equi infection were evaluated. Compared to control horses, horses infected with T. equi had significantly higher SOD1 activities (P <0.05) and PHb (P <0.01), NO2(-) (P <0.001), NOx (P <0.05) and MDA concentrations (P <0.001), and significantly lower EH (P <0.001), CAT (P <0.01) and PON1 (P <0.001) activities, and thiol group concentrations (P <0.05). The comet assay demonstrated significantly increased DNA damage in T. equi infected cells compared to non-infected cells (P <0.001). Infected horses had significantly increased LDH5 isoenzyme activities (P <0.05). There was higher production of ROS/RNS in T. equi-infected horses, which resulted in changes in osmotic fragility, damage to lipids, proteins and DNA, haemolysis and hepatocellular damage. Oxidative stress in horses naturally infected with T. equi could contribute to the pathogenesis of the infection. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Prescottia equi gen. nov., comb. nov.: a new home for an old pathogen.

    PubMed

    Jones, Amanda L; Sutcliffe, Iain C; Goodfellow, Michael

    2013-03-01

    The taxonomic status of Rhodococcus equi, originally isolated from foal specimens, has been the subject of discussion for a number of years. The chequered history of the taxon has prompted this polyphasic analysis of R. equi strains, close members of the genus Rhodococcus and representatives of other genera classified in the order Corynebacteriales, to establish the taxonomic position of this taxon. Thirty one R. equi strains, including the type strain, were examined for genotypic and numerical taxonomic properties. The resultant data are consistent with their classification in the order Corynebacteriales but the R. equi strains formed a distinct phyletic clade away from representatives of other members of the genus Rhodococcus in the 16S rRNA gene tree. Representatives of this clade shared their highest pairwise 16S rRNA gene sequence similarities with the type strain of Rhodococcus kunmingensis (95.2-98.1 %). However, the R. equi taxon was readily distinguished from R. kunmingensis and from the other members of the order Corynebacteriales using a combination of genotypic, chemotypic and phenotypic properties. On the basis of these data the R. equi strains are considered to represent a new genus. The name proposed for this taxon is Prescottia gen. nov., with Prescottia equi comb. nov. as the type species containing the type strain, C 7(T) (= ATCC 25729(T) = ATCC 6939(T) = CCUG 892(T) = CIP 54.72(T) = DSM 20307(T) = HAMBI 2061(T) = NBRC 14956(T) = JCM 1311(T) = JCM 3209(T) = LMG 18452(T) = NBRC 101255(T) = NCTC 1621(T) = NRRL B-16538(T) = VKM Ac-953(T)).

  8. Ivermectin failure in the control of Oxyuris equi in a herd of ponies in France.

    PubMed

    Sallé, Guillaume; Cortet, Jacques; Koch, Christine; Gascogne, Thierry; Reigner, Fabrice; Cabaret, Jacques

    2016-10-15

    Drug resistance in equine gastro-intestinal parasitic nematodes has been reported throughout the world. While the focus is usually put on cyathostomins, observations of macrocylic lactone failure against Oxyuris equi have accumulated over the last decade. Here we report the failure of ivermectin in the control of O. equi in an experimental Welsh pony herd. In a first trial, 6 ponies previously drenched with moxidectin and showing patent O. equi infections were administered ivermectin and subsequently monitored for O. equi egg excretion over one month. This trial demonstrated a failure of ivermectin to control O. equi egg excretion as half of ponies demonstrated recurrent egg excretion in the peri-anal region during 21days after treatment. One year later, six female Welsh ponies drenched with moxidectin demonstrated signs of itching and scratching in their peri-anal region with worms being found transiently in fecal materials three weeks later. Ponies were allocated to three treatment groups, i.e. ivermectin, pyrantel embonate and fenbendazole and monitored for egg excretion over five weeks. Fenbendazole and pyrantel embonate broke ivermectin suboptimal efficacy as soon as 8 and 14days respectively after treatment, while egg excretion remained constant throughout the 41-day long trial in the ivermectin-treated ponies. This is the first report of ivermectin failure against O. equi in France. In the absence of critical efficacy test, it remains unclear whether true resistance is at stake or if these observations confound a constitutive suboptimal efficacy of ivermectin against O. equi. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Influence of Plasmid Type on the Replication of Rhodococcus equi in Host Macrophages

    PubMed Central

    Willingham-Lane, Jennifer M.; Berghaus, Londa J.; Giguère, Steeve

    2016-01-01

    ABSTRACT The soil-dwelling, saprophytic actinomycete Rhodococcus equi is a multihost, facultative intracellular pathogen of macrophages. When inhaled by susceptible foals, it causes severe bronchopneumonia. It is also a pathogen of pigs, which may develop submaxillary lymphadenitis upon exposure. R. equi isolates obtained from foals and pigs possess conjugative plasmids housing a pathogenicity island (PAI) containing a novel family of genes of unknown function called the virulence-associated protein or vap family. The PAI regions of the equine and swine plasmids differ in vap gene composition, with equine isolates possessing six vap genes, including the major virulence determinant vapA, while the PAIs of swine isolates house vapB and five other unique vap genes. Possession of the pVAPA-type virulence plasmid by equine isolates bestows the capacity for intramacrophage replication essential for disease development in vivo. Swine isolates of R. equi are largely unstudied. Here, we show that R. equi isolates from pigs, carrying pVAPB-type plasmids, are able to replicate in a plasmid-dependent manner in macrophages obtained from a variety of species (murine, swine, and equine) and anatomical locations. Similarly, equine isolates carrying pVAPA-type plasmids are capable of replication in swine macrophages. Plasmid swapping between equine and swine strains through conjugation did not alter the intracellular replication capacity of the parental strain, indicating that coevolution of the plasmid and chromosome is not crucial for this attribute. These results demonstrate that while distinct plasmid types exist among R. equi isolates obtained from equine and swine sources, this tropism is not determined by host species-specific intramacrophage replication capabilities. IMPORTANCE This work greatly advances our understanding of the opportunistic pathogen Rhodococcus equi, a disease agent of animals and immunocompromised people. Clinical isolates from diseased foals carry a

  10. Influence of Plasmid Type on the Replication of Rhodococcus equi in Host Macrophages.

    PubMed

    Willingham-Lane, Jennifer M; Berghaus, Londa J; Giguère, Steeve; Hondalus, Mary K

    2016-01-01

    The soil-dwelling, saprophytic actinomycete Rhodococcus equi is a multihost, facultative intracellular pathogen of macrophages. When inhaled by susceptible foals, it causes severe bronchopneumonia. It is also a pathogen of pigs, which may develop submaxillary lymphadenitis upon exposure. R. equi isolates obtained from foals and pigs possess conjugative plasmids housing a pathogenicity island (PAI) containing a novel family of genes of unknown function called the virulence-associated protein or vap family. The PAI regions of the equine and swine plasmids differ in vap gene composition, with equine isolates possessing six vap genes, including the major virulence determinant vapA, while the PAIs of swine isolates house vapB and five other unique vap genes. Possession of the pVAPA-type virulence plasmid by equine isolates bestows the capacity for intramacrophage replication essential for disease development in vivo. Swine isolates of R. equi are largely unstudied. Here, we show that R. equi isolates from pigs, carrying pVAPB-type plasmids, are able to replicate in a plasmid-dependent manner in macrophages obtained from a variety of species (murine, swine, and equine) and anatomical locations. Similarly, equine isolates carrying pVAPA-type plasmids are capable of replication in swine macrophages. Plasmid swapping between equine and swine strains through conjugation did not alter the intracellular replication capacity of the parental strain, indicating that coevolution of the plasmid and chromosome is not crucial for this attribute. These results demonstrate that while distinct plasmid types exist among R. equi isolates obtained from equine and swine sources, this tropism is not determined by host species-specific intramacrophage replication capabilities. IMPORTANCE This work greatly advances our understanding of the opportunistic pathogen Rhodococcus equi, a disease agent of animals and immunocompromised people. Clinical isolates from diseased foals carry a

  11. Corynebacterium equi Infections in Horses, 1958-1984: A Review of 131 Cases

    PubMed Central

    Zink, M. Christine; Yager, Julie A.; Smart, Nonie L.

    1986-01-01

    Of 131 cases of Corynebacterium equi infection in horses submitted for necropsy to the Ontario Veterinary College or Veterinary Laboratory Services, OMAF, Guelph, Ontario from 1958 to 1984, 115 were diagnosed as suppurative pneumonia, and of these 55 had associated ulcerative enterocolitis. Only five animals had intestinal involvement without pulmonary lesions. The remaining 11 cases included arthritis/cellulitis, skin abscesses and submandibular lymphadenitis. While the lung, intestine and associated lymph nodes yielded C. equi most frequently, in 21% of cases C. equi was also cultured from parenchymatous organs (spleen, liver or kidney) or blood. Corynebacterium equi infection accounted for 10% of all foals submitted for postmortem examination and 45% of all foals with pneumonia. Affected foals were one to four months of age. Submissions occurred between the months of May and August with a peak during July. There was a significantly greater prevalence of C. equi infection in Standardbreds when compared with other breeds. Of foals in this study, 36% were from farms which had had other horses succumb to this disease. Of the foals with pulmonary involvement, 21% did not have fever or clinical signs referable to the respiratory or gastrointestinal systems, findings which indicated that a large percentage of cases were subclinical. PMID:17422658

  12. Conjugal Transfer of a Virulence Plasmid in the Opportunistic Intracellular Actinomycete Rhodococcus equi

    PubMed Central

    Tripathi, V. N.; Harding, W. C.; Willingham-Lane, J. M.

    2012-01-01

    Rhodococcus equi is a facultative intracellular, Gram-positive, soilborne actinomycete which can cause severe pyogranulomatous pneumonia with abscessation in young horses (foals) and in immunocompromised people, such as persons with AIDS. All strains of R. equi isolated from foals and approximately a third isolated from humans contain a large, ∼81-kb plasmid which is essential for the intramacrophage growth of the organism and for virulence in foals and murine in vivo model systems. We found that the entire virulence plasmid could be transferred from plasmid-containing strains of R. equi (donor) to plasmid-free R. equi strains (recipient) at a high frequency and that plasmid transmission reestablished the capacity for intracellular growth in macrophages. Plasmid transfer required living cells and cell-to-cell contact and was unaffected by the presence of DNase, factors pointing to conjugation as the major means of genetic transfer. Deletion of a putative relaxase-encoding gene, traA, located in the proposed conjugative region of the plasmid, abolished plasmid transfer. Reversion of the traA mutation restored plasmid transmissibility. Finally, plasmid transmission to other Rhodococcus species and some additional related organisms was demonstrated. This is the first study showing a virulence plasmid transfer in R. equi, and it establishes a mechanism by which the virulence plasmid can move among bacteria in the soil. PMID:23042997

  13. The sensor kinase MprB is required for Rhodococcus equi virulence.

    PubMed

    MacArthur, Iain; Parreira, Valeria R; Lepp, Dion; Mutharia, Lucy M; Vazquez-Boland, José A; Prescott, John F

    2011-01-10

    Rhodococcus equi is a soil bacterium and, like Mycobacterium tuberculosis, a member of the mycolata. Through possession of a virulence plasmid, it has the ability to infect the alveolar macrophages of foals, resulting in pyogranulomatous bronchopneumonia. The virulence plasmid has an orphan two-component system (TCS) regulatory gene, orf8, mutation of which completely attenuates virulence. This study attempted to find the cognate sensor kinase (SK) of orf8. Annotation of the R. equi strain 103 genome identified 23 TCSs encoded on the chromosome, which were used in a DNA microarray to compare TCS gene transcription in murine macrophage-like cells to growth in vitro. This identified six SKs as significantly up-regulated during growth in macrophages. Mutants of these SKs were constructed and their ability to persist in macrophages was determined with one SK, MprB, found to be required for intracellular survival. The attenuation of the mprB- mutant, and its complementation, was confirmed in a mouse virulence assay. In silico analysis of the R. equi genome sequence identified an MprA binding box motif homologous to that of M. tuberculosis, on mprA, pepD, sigB and sigE. The results of this study also show that R. equi responds to the macrophage environment differently from M. tuberculosis. MprB is the first SK identified as required for R. equi virulence and intracellular survival. Copyright © 2010 Elsevier B.V. All rights reserved.

  14. Conjugal transfer of a virulence plasmid in the opportunistic intracellular actinomycete Rhodococcus equi.

    PubMed

    Tripathi, V N; Harding, W C; Willingham-Lane, J M; Hondalus, M K

    2012-12-01

    Rhodococcus equi is a facultative intracellular, Gram-positive, soilborne actinomycete which can cause severe pyogranulomatous pneumonia with abscessation in young horses (foals) and in immunocompromised people, such as persons with AIDS. All strains of R. equi isolated from foals and approximately a third isolated from humans contain a large, ~81-kb plasmid which is essential for the intramacrophage growth of the organism and for virulence in foals and murine in vivo model systems. We found that the entire virulence plasmid could be transferred from plasmid-containing strains of R. equi (donor) to plasmid-free R. equi strains (recipient) at a high frequency and that plasmid transmission reestablished the capacity for intracellular growth in macrophages. Plasmid transfer required living cells and cell-to-cell contact and was unaffected by the presence of DNase, factors pointing to conjugation as the major means of genetic transfer. Deletion of a putative relaxase-encoding gene, traA, located in the proposed conjugative region of the plasmid, abolished plasmid transfer. Reversion of the traA mutation restored plasmid transmissibility. Finally, plasmid transmission to other Rhodococcus species and some additional related organisms was demonstrated. This is the first study showing a virulence plasmid transfer in R. equi, and it establishes a mechanism by which the virulence plasmid can move among bacteria in the soil.

  15. Corynebacterium equi Infections in Horses, 1958-1984: A Review of 131 Cases.

    PubMed

    Zink, M C; Yager, J A; Smart, N L

    1986-05-01

    Of 131 cases of Corynebacterium equi infection in horses submitted for necropsy to the Ontario Veterinary College or Veterinary Laboratory Services, OMAF, Guelph, Ontario from 1958 to 1984, 115 were diagnosed as suppurative pneumonia, and of these 55 had associated ulcerative enterocolitis. Only five animals had intestinal involvement without pulmonary lesions. The remaining 11 cases included arthritis/cellulitis, skin abscesses and submandibular lymphadenitis. While the lung, intestine and associated lymph nodes yielded C. equi most frequently, in 21% of cases C. equi was also cultured from parenchymatous organs (spleen, liver or kidney) or blood. Corynebacterium equi infection accounted for 10% of all foals submitted for postmortem examination and 45% of all foals with pneumonia. Affected foals were one to four months of age. Submissions occurred between the months of May and August with a peak during July. There was a significantly greater prevalence of C. equi infection in Standardbreds when compared with other breeds. Of foals in this study, 36% were from farms which had had other horses succumb to this disease. Of the foals with pulmonary involvement, 21% did not have fever or clinical signs referable to the respiratory or gastrointestinal systems, findings which indicated that a large percentage of cases were subclinical.

  16. Assessment of theileria equi and babesia caballi infections in equine populations in Egypt by molecular, serological and hematological approaches

    USDA-ARS?s Scientific Manuscript database

    Background: Equine piroplasmosis caused by Theileria equi, Babesia caballi, or both, cause significant economic losses in the equine industry and remains uncontrolled in Egypt. Methods: T. equi and B. caballi infections were assessed in blood from 88 horses and 51 donkeys from different localities ...

  17. 75 FR 25234 - EquiPower Resources Management, LLC; Supplemental Notice That Initial Market-Based Rate Filing...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-07

    ... Energy Regulatory Commission EquiPower Resources Management, LLC; Supplemental Notice That Initial Market... supplemental notice in the above-referenced proceeding of EquiPower Resources Management, LLC's application for market-based rate authority, with an accompanying rate ] tariff, noting that such application includes...

  18. Protective Effects of Passively Transferred Merozoite-Specific Antibodies against Theileria equi in Horses with Severe Combined Immunodeficiency

    PubMed Central

    Kappmeyer, Lowell S.; Ueti, Massaro W.; Wagner, Bettina; Knowles, Donald P.

    2012-01-01

    Theileria equi immune plasma was infused into young horses (foals) with severe combined immunodeficiency. Although all foals became infected following intravenous challenge with homologous T. equi merozoite stabilate, delayed time to peak parasitemia occurred. Protective effects were associated with a predominance of passively transferred merozoite-specific IgG3. PMID:22038847

  19. Effect of prophylactic administration of hyperimmune plasma to prevent Rhodococcus equi infection on foals from endemically affected farms.

    PubMed

    Higuchi, T; Arakawa, T; Hashikura, S; Inui, T; Senba, H; Takai, S

    1999-11-01

    The effect on foals of prophylactic administration of hyperimmune plasma to prevent R. equi infection was investigated on three farms at which R. equi infection was endemic. Sixteen foals between 10 and 39 days of age were intravenously given 1-21 of hyperimmune plasma. ELISA antibody titres against R. equi were significantly increased and maintained at high levels for over 30 days in most of the recipient foals. The prevalence of R. equi infection was 6.3% (1/16) in the foals that received the immune plasma, and 26.3% (5/19) in the control foals not given the immune plasma on the three farms. For 2 years before and after this field trial on the three farms, 18 of 64 foals (28.1%) showed clinical signs of respiratory tract infection and four of them died of R. equi pneumonia. Heavy contamination of horses and their environment with virulent R. equi was detected by colony blotting, and plasmid profiles also suggested that foals on the three farms were constantly exposed to virulent R. equi. The results of this field trial support previous observations by some researchers that the administration of hyperimmune plasma to foals in the early days of life promotes prevention of R. equi infection on endemic farms; however, the mechanism of hyperimmune plasma protection remains unclear.

  20. Effect of hyperimmune plasma on the severity of pneumonia caused by Rhodococcus equi in experimentally infected foals.

    PubMed

    Caston, Stephanie S; McClure, Scott R; Martens, Ronald J; Chaffin, M Keith; Miles, Kristina G; Griffith, Ronald W; Cohen, Noah D

    2006-01-01

    This study evaluated the prophylactic effectiveness of hyperimmune plasma (HIP) as an aid in the prevention of pneumonia caused by experimental infection with Rhodococcus equi. Thirty neonatal foals were administered R. equi HIP or saline at 2 days of age and were infected with virulent R. equi at 7 days. All foals developed signs or symptoms of respiratory disease. Radiographic scores on day 28 and neutrophil concentrations on day 49 were significantly greater in control foals, and time to respiratory effort score of 2 or higher was significantly shorter for control foals. Three foals, all in the principal group, died or were euthanized before the end of the study, but there was no significant difference in mortality between groups. VapA titers were significantly greater in principal foals. Administration of R. equi HIP decreased the severity of radiographic lesions and prolonged time to increased respiratory effort due to R. equi-induced pneumonia.

  1. Efficacy of alphacypermetrin pour-on against natural Werneckiella equi infestation on donkeys (Equus asinus).

    PubMed

    Veneziano, Vincenzo; Neglia, Gianluca; Galietti, Alfredo; Rufrano, Domenico; Bassini, Andrea; Mariani, Ugo; Gokbulut, Cengiz

    2012-09-01

    The chewing louse Werneckiella equi is an ectoparasite of donkeys and other equids. Alphacypermethrin (ACYP) is a pyrethroid insecticide commonly used for the control of insects of veterinary and public health concerns. A trial was conducted to assess the efficacy of ACYP against W. equi on naturally infested donkeys. Parasitological investigations were performed on 13 animals. On day 0, the donkeys received ACYP pour-on at the manufacturer's recommended dose rate for cattle. Louse counts were performed on days -1, 7, 14, 21, 28, 35, 42, 49, and 56 at seven predilection sites on the skin of each donkey. ACYP was completely effective (100 %) from day 7, until the end of the study. ACYP was well tolerated by all animals as there were no observed clinical adverse reactions. ACYP was highly effective, safe, user-friendly, and considered to be suitable for the treatment of donkeys for W. equi infestation.

  2. Equid herpesvirus 1 and rhodococcus equi coinfection in a foal with bronchointerstitial pneumonia.

    PubMed

    Perez-Ecija, Alejandro; Mendoza, Francisco Javier; Estepa, José Carlos; Bautista, María José; Pérez, José

    2016-10-01

    A 2-month-old foal with septic shock and severe respiratory distress was referred to the Veterinary Teaching Hospital. Due to poor prognosis, the foal was euthanized. Histopathology showed lesions suggestive of Rhodococcus equi infection associated with a diffuse interstitial infiltrate of foamy macrophages and syncytial cells presenting large acidophilic intranuclear inclusion bodies, fibrin exudates and hyaline membranes. Bacteriological examination from lung and respiratory exudates confirmed R. equi infection, whilst immunohistochemistry and PCR yielded a positive result for Equid herpesvirus type 1 (EHV-1). Several etiologies have been proposed for bronchointerstitial pneumonia in foals, although a multifactorial origin for this lesional pattern could be possible. This work is the first one describing a combined EHV-1 and R. equi infection in a foal affected with bronchointerstitial pneumonia.

  3. Rhodococcus equi human clinical isolates enter and survive within human alveolar epithelial cells.

    PubMed

    Ramos-Vivas, J; Pilares-Ortega, L; Remuzgo-Martínez, S; Padilla, D; Gutiérrez-Díaz, J L; Navas-Méndez, J

    2011-05-01

    Rhodococcus equi is an emerging opportunistic human pathogen associated with immunosuppressed people, especially those infected with the human immunodeficiency virus (HIV). This pathogen resides primarily within lung macrophages of infected patients, which may explain in part its ability to escape normal pulmonary defense mechanisms. Despite numerous studies as a pulmonary pathogen in foals, where a plasmid seems to play an important role in virulence, information on the pathogenesis of this pathogen in humans is still scarce. In this study, fluorescence microscopy and vancomycin protection assays were used to investigate the ability of R. equi human isolates to adhere to and to invade the human alveolar epithelial cell line A549. Our findings indicate that some R. equi clinical strains are capable of adhering, entering and surviving within the alveolar cell line, which may contribute to the pathogen persistence in lung tissues. Copyright © 2011 Institut Pasteur. Published by Elsevier SAS. All rights reserved.

  4. Equid herpesvirus 1 and rhodococcus equi coinfection in a foal with bronchointerstitial pneumonia

    PubMed Central

    PEREZ-ECIJA, Alejandro; MENDOZA, Francisco Javier; ESTEPA, José Carlos; BAUTISTA, María José; PÉREZ, José

    2016-01-01

    A 2-month-old foal with septic shock and severe respiratory distress was referred to the Veterinary Teaching Hospital. Due to poor prognosis, the foal was euthanized. Histopathology showed lesions suggestive of Rhodococcus equi infection associated with a diffuse interstitial infiltrate of foamy macrophages and syncytial cells presenting large acidophilic intranuclear inclusion bodies, fibrin exudates and hyaline membranes. Bacteriological examination from lung and respiratory exudates confirmed R. equi infection, whilst immunohistochemistry and PCR yielded a positive result for Equid herpesvirus type 1 (EHV-1). Several etiologies have been proposed for bronchointerstitial pneumonia in foals, although a multifactorial origin for this lesional pattern could be possible. This work is the first one describing a combined EHV-1 and R. equi infection in a foal affected with bronchointerstitial pneumonia. PMID:27264610

  5. The Antiphagocytic Activity of SeM of Streptococcus equi Requires Capsule.

    PubMed

    Timoney, John F; Suther, Pranav; Velineni, Sridhar; Artiushin, Sergey C

    2014-01-01

    Resistance to phagocytosis is a crucial virulence property of Streptococcus equi (Streptococcus equi subsp. equi; Se), the cause of equine strangles. The contribution and interdependence of capsule and SeM to killing in equine blood and neutrophils were investigated in naturally occurring strains of Se. Strains CF32, SF463 were capsule and SeM positive, strains Lex90, Lex93 were capsule negative and SeM positive and strains Se19, Se1-8 were capsule positive and SeM deficient. Phagocytosis and killing of Se19, Se1-8, Lex90 and Lex93 in equine blood and by neutrophils suspended in serum were significantly (P ≤ 0.02) greater compared to CF32 and SF463. The results indicate capsule and SeM are both required for resistance to phagocytosis and killing and that the anti-phagocytic property of SeM is greatly reduced in the absence of capsule.

  6. Plasmid Profiles of Virulent Rhodococcus equi Strains Isolated from Infected Foals in Poland.

    PubMed

    Kalinowski, Marcin; Grądzki, Zbigniew; Jarosz, Łukasz; Kato, Kiyoko; Hieda, Yu; Kakuda, Tsutomu; Takai, Shinji

    2016-01-01

    Rhodococcus equi is an important bacterial pathogen in foals up to 6 months old, widespread in horse farms all over the world. It was found that only virulent R. equi strains expressing 15-17 kDa virulence-associated protein (VapA) and having large virulence plasmid of 85-90 kb containing vapA gene are pathogenic for horses. To date, 12 plasmid types have been reported in VapA positive strains from horses. There are no data concerning plasmid types of Polish field R. equi strains isolated from horses and horse farm environment. The aim of the study is to determine plasmid profiles of virulent R. equi strains isolated in Poland from dead foals as well as from soil samples taken from horse breeding farms. Plasmid profiles of 10 clinical strains derived from 8 farms and 11 environmental strains from 3 farms, confirmed as virulent by PCR, were compared with 12 reference strains containing the known plasmid size and type. Plasmid DNAs were analysed by digestion with the restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII for detailed comparison and estimation of plasmid sizes. The results of RFLP analysis revealed that all except one isolates used in the study are classified as VapA 85 kb type I plasmid. One strain harboured VapA 87 kb type I plasmid. This is the first report of plasmid types of Polish field R. equi strains. The results of our preliminary investigations on horse farms located in central and eastern Poland indicate that the virulent R. equi strains thus far isolated from diseased foals and horse farms environment represent a highly uniform plasmid pattern.

  7. Identification of genomic loci associated with Rhodococcus equi susceptibility in foals.

    PubMed

    McQueen, Cole M; Doan, Ryan; Dindot, Scott V; Bourquin, Jessica R; Zlatev, Zlatomir Z; Chaffin, M Keith; Blodgett, Glenn P; Ivanov, Ivan; Cohen, Noah D

    2014-01-01

    Pneumonia caused by Rhodococcus equi is a common cause of disease and death in foals. Although agent and environmental factors contribute to the incidence of this disease, the genetic factors influencing the clinical outcomes of R. equi pneumonia are ill-defined. Here, we performed independent single nucleotide polymorphism (SNP)- and copy number variant (CNV)-based genome-wide association studies to identify genomic loci associated with R. equi pneumonia in foals. Foals at a large Quarter Horse breeding farm were categorized into 3 groups: 1) foals with R. equi pneumonia (clinical group [N = 43]); 2) foals with ultrasonographic evidence of pulmonary lesions that never developed clinical signs of pneumonia (subclinical group [N = 156]); and, 3) foals without clinical signs or ultrasonographic evidence of pneumonia (unaffected group [N = 49]). From each group, 24 foals were randomly selected and used for independent SNP- and CNV-based genome-wide association studies (GWAS). The SNP-based GWAS identified a region on chromosome 26 that had moderate evidence of association with R. equi pneumonia when comparing clinical and subclinical foals. A joint analysis including all study foals revealed a 3- to 4-fold increase in odds of disease for a homozygous SNP within the associated region when comparing the clinical group with either of the other 2 groups of foals or their combination. The region contains the transient receptor potential cation channel, subfamily M, member 2 (TRPM2) gene, which is involved in neutrophil function. No associations were identified in the CNV-based GWAS. Collectively, these data identify a region on chromosome 26 associated with R. equi pneumonia in foals, providing evidence that genetic factors may indeed contribute to this important disease of foals.

  8. The Hydroxamate Siderophore Rhequichelin Is Required for Virulence of the Pathogenic Actinomycete Rhodococcus equi

    PubMed Central

    Coulson, Garry B.; Miranda-CasoLuengo, Aleksandra; Vázquez-Boland, José A.; Hondalus, Mary K.

    2012-01-01

    We previously showed that the facultative intracellular pathogen Rhodococcus equi produces a nondiffusible and catecholate-containing siderophore (rhequibactin) involved in iron acquisition during saprophytic growth. Here, we provide evidence that the rhbABCDE cluster directs the biosynthesis of a hydroxamate siderophore, rhequichelin, that plays a key role in virulence. The rhbC gene encodes a nonribosomal peptide synthetase that is predicted to produce a tetrapeptide consisting of N5-formyl-N5-hydroxyornithine, serine, N5-hydroxyornithine, and N5-acyl-N5-hydroxyornithine. The other rhb genes encode putative tailoring enzymes mediating modification of ornithine residues incorporated into the hydroxamate product of RhbC. Transcription of rhbC was upregulated during growth in iron-depleted medium, suggesting that it plays a role in iron acquisition. This was confirmed by deletion of rhbCD, rendering the resulting strain R. equi SID2 unable to grow in the presence of the iron chelator 2,2-dipyridyl. Supernatant of the wild-type strain rescued the phenotype of R. equi SID2. The importance of rhequichelin in virulence was highlighted by the rapid increase in transcription levels of rhbC following infection and the inability of R. equi SID2 to grow within macrophages. Unlike the wild-type strain, R. equi SID2 was unable to replicate in vivo and was rapidly cleared from the lungs of infected mice. Rhequichelin is thus a key virulence-associated factor, although nonpathogenic Rhodococcus species also appear to produce rhequichelin or a structurally closely related compound. Rhequichelin biosynthesis may therefore be considered an example of cooption of a core actinobacterial trait in the evolution of R. equi virulence. PMID:22966042

  9. Plasmid Profiles of Virulent Rhodococcus equi Strains Isolated from Infected Foals in Poland

    PubMed Central

    Kalinowski, Marcin; Grądzki, Zbigniew; Jarosz, Łukasz; Kato, Kiyoko; Hieda, Yu; Kakuda, Tsutomu; Takai, Shinji

    2016-01-01

    Rhodococcus equi is an important bacterial pathogen in foals up to 6 months old, widespread in horse farms all over the world. It was found that only virulent R. equi strains expressing 15–17 kDa virulence-associated protein (VapA) and having large virulence plasmid of 85–90 kb containing vapA gene are pathogenic for horses. To date, 12 plasmid types have been reported in VapA positive strains from horses. There are no data concerning plasmid types of Polish field R. equi strains isolated from horses and horse farm environment. The aim of the study is to determine plasmid profiles of virulent R. equi strains isolated in Poland from dead foals as well as from soil samples taken from horse breeding farms. Plasmid profiles of 10 clinical strains derived from 8 farms and 11 environmental strains from 3 farms, confirmed as virulent by PCR, were compared with 12 reference strains containing the known plasmid size and type. Plasmid DNAs were analysed by digestion with the restriction endonucleases BamHI, EcoRI, EcoT22I, and HindIII for detailed comparison and estimation of plasmid sizes. The results of RFLP analysis revealed that all except one isolates used in the study are classified as VapA 85 kb type I plasmid. One strain harboured VapA 87 kb type I plasmid. This is the first report of plasmid types of Polish field R. equi strains. The results of our preliminary investigations on horse farms located in central and eastern Poland indicate that the virulent R. equi strains thus far isolated from diseased foals and horse farms environment represent a highly uniform plasmid pattern. PMID:27074033

  10. The hydroxamate siderophore rhequichelin is required for virulence of the pathogenic actinomycete Rhodococcus equi.

    PubMed

    Miranda-Casoluengo, Raúl; Coulson, Garry B; Miranda-Casoluengo, Aleksandra; Vázquez-Boland, José A; Hondalus, Mary K; Meijer, Wim G

    2012-12-01

    We previously showed that the facultative intracellular pathogen Rhodococcus equi produces a nondiffusible and catecholate-containing siderophore (rhequibactin) involved in iron acquisition during saprophytic growth. Here, we provide evidence that the rhbABCDE cluster directs the biosynthesis of a hydroxamate siderophore, rhequichelin, that plays a key role in virulence. The rhbC gene encodes a nonribosomal peptide synthetase that is predicted to produce a tetrapeptide consisting of N(5)-formyl-N(5)-hydroxyornithine, serine, N(5)-hydroxyornithine, and N(5)-acyl-N(5)-hydroxyornithine. The other rhb genes encode putative tailoring enzymes mediating modification of ornithine residues incorporated into the hydroxamate product of RhbC. Transcription of rhbC was upregulated during growth in iron-depleted medium, suggesting that it plays a role in iron acquisition. This was confirmed by deletion of rhbCD, rendering the resulting strain R. equi SID2 unable to grow in the presence of the iron chelator 2,2-dipyridyl. Supernatant of the wild-type strain rescued the phenotype of R. equi SID2. The importance of rhequichelin in virulence was highlighted by the rapid increase in transcription levels of rhbC following infection and the inability of R. equi SID2 to grow within macrophages. Unlike the wild-type strain, R. equi SID2 was unable to replicate in vivo and was rapidly cleared from the lungs of infected mice. Rhequichelin is thus a key virulence-associated factor, although nonpathogenic Rhodococcus species also appear to produce rhequichelin or a structurally closely related compound. Rhequichelin biosynthesis may therefore be considered an example of cooption of a core actinobacterial trait in the evolution of R. equi virulence.

  11. Foal Monocyte-Derived Dendritic Cells Become Activated upon Rhodococcus equi Infection▿ †

    PubMed Central

    Flaminio, M. Julia B. F.; Nydam, Daryl V.; Marquis, Hélène; Matychak, Mary Beth; Giguère, Steeve

    2009-01-01

    Susceptibility of foals to Rhodococcus equi pneumonia is exclusive to the first few months of life. The objective of this study was to investigate the immediate immunologic response of foal and adult horse antigen-presenting cells (APCs) upon infection with R. equi. We measured the activation of the antigen-presenting major histocompatibility complex (MHC) class II molecule, costimulatory molecules CD40 and CD86, the cytokine interleukin-12 (IL-12), and the transcriptional factor interferon regulatory factor 1 (IRF-1) in monocyte-derived macrophages (mMOs) and dendritic cells (mDCs) of adult horses and foals of different ages (from birth to 3 months of age) infected with virulent R. equi or its avirulent, plasmid-cured derivative. Infection with virulent or avirulent R. equi induced (P ≤ 0.01) the expression of IL-12p35 and IL-12p40 mRNAs in foal mMOs and mDCs at different ages. This response was likely mediated by the higher (P = 0.008) expression of IRF-1 in foal mDCs at birth than in adult horse mDCs. R. equi infection promoted comparable expression of costimulatory molecules CD86 and CD40 in foal and adult horse cells. The cytokine and costimulatory response by foal mDCs was not accompanied by robust MHC class II molecule expression. These data suggest that foal APCs detect the presence of R. equi and respond with the expression of the Th1-inducing cytokine IL-12. Nevertheless, there seems to be a limitation to MHC class II molecule expression which we hypothesize may compromise the efficient priming of naïve effector cells in early life. PMID:19109450

  12. Efficacy of pyrantel pamoate and ivermectin paste formulations against naturally acquired Oxyuris equi infections in horses.

    PubMed

    Reinemeyer, Craig R; Prado, Julio C; Nichols, Eric C; Marchiondo, Alan A

    2010-07-15

    In recent years, numerous veterinary practitioners have reported anecdotal episodes in which anthelmintic treatment did not appear to deliver the expected efficacy against equine pinworms (Oxyuris equi). Anthelmintic resistance has not been demonstrated formally in equine pinworms, so a clinical study was designed to evaluate the efficacy of paste formulations of pyrantel pamoate or ivermectin against naturally acquired infections with O. equi. Twenty-one horses (>4 months to 15 years of age) with patent, naturally acquired pinworm infections were blocked by source of origin and allocated randomly to one of three treatment groups: horses (n=7) assigned to Group 1 were treated orally with pyrantel pamoate paste at a dosage of 13.2 mg/kg (2x label dosage), Group 2 horses (n=7) were untreated controls, and horses (n=7) assigned to Group 3 were treated orally with ivermectin paste at a dosage of 200 microg/kg. Fourteen days after treatment, horses were euthanatized, necropsied, and large intestinal contents were processed for recovery of adult pinworms. In addition, duplicate 1% aliquots of intestinal contents from the cecum, ventral colon, dorsal colon, and small colon were collected, preserved, and examined for recovery and enumeration of fourth-stage larval O. equi. Anthelmintic efficacy against pinworms was evaluated by comparing the post-treatment worm counts of Groups 1 and 3 to those of control animals. Mean numbers of O. equi adults recovered postmortem were significantly decreased by both pyrantel pamoate (P=0.0366) and ivermectin (P=0.0137) treatment, with respective efficacies of 91.2% and 96.0%. In addition, both products demonstrated >99% efficacy against fourth-stage O. equi larvae. The current study demonstrated acceptable adulticidal and larvicidal efficacy of both pyrantel pamoate and ivermectin paste formulations against O. equi and did not support the existence of macrocyclic lactone or pyrimidine resistance in the pinworm populations evaluated.

  13. Klossiella equi in a donkey--a first case report from Iran.

    PubMed

    Rezaie, A; Bahrami, S; Ansari, M

    2013-09-01

    Klossiella equi is the only known and rarely reported coccidian parasite of the renal paranchyma of equids. An aged male donkey (Equus asinus asinus) was submitted to necropsy department of veterinary hospital. In histopathological study of renal sections different developmental stages of parasite were observed. These stages were as follow: Trophozoites, microgametes, macrogametes, sporont, budding sporont, sporoblasts, free sporoblasts, mature sporoblast and sporocyst. Parasitic infection with K. equi was encountered in the donkey. According to literature review this is the first report of donkey klossiellosis in Iran.

  14. Natural Infection of the South American Tapir ( Tapirus terrestris ) by Theileria equi.

    PubMed

    Da Silveira, Alexandre Welzel; De Oliveira, Gustavo Gomes; Menezes Santos, Leandro; da Silva Azuaga, Lucas Bezerra; Macedo Coutinho, Claudia Regina; Echeverria, Jessica Teles; Antunes, Tamires Ramborger; do Nascimento Ramos, Carlos Alberto; Izabel de Souza, Alda

    2017-04-01

    Theileria equi is a tick-borne piroplasm considered endemic in equines in Brazil. The cohabitation of domestic and wild animals in areas of extensive cattle breeding favors the close contact between different species and the sharing of vectors and, consequently, pathogens. We report the natural infection of a young South American tapir ( Tapirus terrestris ) by T. equi in Mato Grosso do Sul, Brazil. Although it was not possible to associate the clinical and hematologic status of the animal with the infection by the protozoan parasite, our report represents an alert on the sharing of pathogens between domestic and wild animals.

  15. Transfer of the virulence-associated protein a-bearing plasmid between field strains of virulent and avirulent Rhodococcus equi.

    PubMed

    Stoughton, W; Poole, T; Kuskie, K; Liu, M; Bishop, K; Morrissey, A; Takai, S; Cohen, N

    2013-01-01

    Virulent and avirulent isolates of Rhodococcus equi coexist in equine feces and the environment and are a source of infection for foals. The extent to which plasmid transfer occurs among field strains is ill-defined and this information is important for understanding the epidemiology of R. equi infections of foals. To estimate the frequency of transfer of the virulence plasmid between virulent and avirulent strains of R. equi derived from foals and their environment. None. In vitro study; 5 rifampin-susceptible, virulent R. equi isolates obtained from clinically affected foals or air samples from a farm with a history of recurrent R. equi foal pneumonia were each mixed with 5 rifampin-resistant, avirulent isolates derived from soil samples, using solid medium, at a ratio of 10 donor cells (virulent) per recipient cell. Presumed transconjugates were detected by plating on media with rifampin and colony immunoblotting to detect the presence of the virulence-associated protein A. Three presumed transconjugates were detected among 2,037 recipient colonies, indicating an overall estimated transfer frequency of 0.15% (95% CI, 0.03–0.43%). All 3 transconjugates were associated with a single donor and 2 recipient strains. Genotyping and multiplex PCR of presumed transconjugates demonstrated transfer of the virulence-associated protein A-bearing plasmid between virulent and avirulent R. equi. Transfer of the virulence plasmid occurs with relatively high frequency. These findings could impact strategies to control or prevent R. equi through environmental management.

  16. Proposal to replace the illegitimate genus name Prescottia Jones et al. 2013 with the genus name Prescottella gen. nov. and to replace the illegitimate combination Prescottia equi Jones et al. 2013 with Prescottella equi comb. nov.

    PubMed

    Jones, Amanda L; Sutcliffe, Iain C; Goodfellow, Michael

    2013-06-01

    Recently we proposed that Rhodococcus equi (Magnusson 1923) Goodfellow and Alderson 1977 be transferred to a novel genus, Prescottia, as Prescottia equi comb. nov. However, in accordance with Principle 2 and Rule 51b(4) of the Bacteriological Code (1990 Revision), the bacterial genus name Prescottia Jones et al. 2013 is deemed illegitimate as this name has been used previously for a plant genus within the family Orchidaceae. Consequently, a new genus name, Prescottella gen. nov. is proposed for the bacterial taxon and a new combination Prescottella equi comb. nov. is proposed for the type species.

  17. Effects of Administration of Live or Inactivated Virulent Rhodococccus equi and Age on the Fecal Microbiome of Neonatal Foals

    PubMed Central

    Bordin, Angela I.; Suchodolski, Jan S.; Markel, Melissa E.; Weaver, Kaytee B.; Steiner, Jörg M.; Dowd, Scot E.; Pillai, Suresh; Cohen, Noah D.

    2013-01-01

    Background Rhodococcus equi is an important pathogen of foals. Enteral administration of live, virulent R. equi during early life has been documented to protect against subsequent intrabronchial challenge with R. equi, indicating that enteral mucosal immunization may be protective. Evidence exists that mucosal immune responses develop against both live and inactivated micro-organisms. The extent to which live or inactivated R. equi might alter the intestinal microbiome of foals is unknown. This is an important question because the intestinal microbiome of neonates of other species is known to change over time and to influence host development. To our knowledge, changes in the intestinal microbiome of foals during early life have not been reported. Thus, the purpose of this study was to determine whether age (during the first month of life) or administration of either live virulent R. equi (at a dose reported to protect foals against subsequent intrabronchial challenge, viz., 1×1010 colony forming units [CFU]) or inactivated virulent R. equi (at higher doses, viz., 2×1010 and 1×1011 [CFU]) altered the fecal microbiome of foals. Methodology/Principal Findings Fecal swab samples from 42 healthy foals after vaccination with low-dose inactivated R. equi (n = 9), high-dose inactivated R. equi (n = 10), live R. equi (n = 6), control with cholera toxin B (CTB, n = 9), and control without CTB (n = 8) were evaluated by 454-pyrosequencing of the 16S rRNA gene and by qPCR. No impact of treatment was observed among vaccinated foals; however, marked and significant differences in microbial communities and diversity were observed between foals at 30 days of age relative to 2 days of age. Conclusions The results suggest age-related changes in the fecal microbial population of healthy foals do occur, however, mucosal vaccination does not result in major changes of the fecal microbiome in foals. PMID:23785508

  18. Molecular and serological detection of Theileria equi and Babesia caballi in donkeys (Equus asinus) in Brazil.

    PubMed

    Machado, R Z; Toledo, C Z P; Teixeira, M C A; André, M R; Freschi, C R; Sampaio, P H

    2012-05-25

    Piroplasmosis in donkeys has been recognized as a serious problem of major economic importance, since the affected animals manifest loss of appetite and decreased working capacity. The present work is aimed at detecting infection or exposure of donkeys in São Paulo, Brazil to Theileria (T.) equi and Babesia (B.) caballi using molecular and serological approaches. EDTA-blood and serum samples were collected from 88 donkeys (Equus asinus). From 88 sampled donkeys, 65 (73.86%; 95% confidence interval, PI=63.41, 82.65) and 82 (93.2%; 95% confidence interval, PI=85.75, 97.46) animals showed IgG antibodies to T. equi (by ELISA) and B. caballi (by IFAT), respectively. Twenty-eight (31.81%; 95% confidence interval, PI=22.3, 42.61) and 18 (20.45%; 95% confidence interval, PI=12.6, 30.39) donkeys were positive to T. equi and B. caballi nested PCR assays, respectively. The results indicated that T. equi and B. caballi are prevalent among donkeys in Brazil.

  19. [Rhodococcus equi infection in AIDS patients: retrospective analysis of 13 patients in Argentina].

    PubMed

    Corti, Marcelo; Solari, Rubén; De Carolis, Luis; Palmieri, Omar; Rollet, Raquel; Shah, Haroun N

    2014-08-01

    Rhodococcus equi is a gram positive coccoid rod that causes pulmonary infections in immunosuppressed patients. We retrospectively analyzed epidemiological, clinical, microbiological, radiological, and immunological features as well as the outcomes of 13 AIDS patients with R. equi infection. Between January 1994 and December 2012, 13 patients attending the AIDS department of the Infectious Diseases reference hospital in Buenos Aires were diagnosed with R. equi infection. All were men, the median age was 27 years. At the time of diagnosis, the median of CD4+ T cell counts was 11 cells/μl Twelve patients presented pulmonary disease with isolation of the microorganism from sputum or bronchoalveolar lavage; in the other patient the diagnosis was postmortem with positive culture of cerebrospinal fluid. The most frequent clinical manifestations were fever, haemoptysis, and weight loss. The predominant radiological finding was lobe consolidation with cavitation. Nine patients died after a median survival of 5.5 months. In all of them, cultures persisted positive until the last admission. The other 4 patients did continue clinical follow-ups. The insidious course of R. equi disease and the difficulties in the isolation of the microorganism contribute to the delay in the diagnosis and to the high mortality rate of this opportunistic infection.

  20. Diversion of phagosome trafficking by pathogenic Rhodococcus equi depends on mycolic acid chain length

    PubMed Central

    Sydor, Tobias; Bargen, Kristine; Hsu, Fong-Fu; Huth, Gitta; Holst, Otto; Wohlmann, Jens; Becken, Ulrike; Dykstra, Tobias; Söhl, Kristina; Lindner, Buko; Prescott, John F; Schaible, Ulrich E; Utermöhlen, Olaf; Haas, Albert

    2013-01-01

    Rhodococcus equi is a close relative of Mycobacterium spp. and a facultative intracellular pathogen which arrests phagosome maturation in macrophages before the late endocytic stage. We have screened a transposon mutant library of R. equi for mutants with decreased capability to prevent phagolysosome formation. This screen yielded a mutant in the gene for β-ketoacyl-(acyl carrier protein)-synthase A (KasA), a key enzyme of the long-chain mycolic acid synthesizing FAS-II system. The longest kasA mutant mycolic acid chains were 10 carbon units shorter than those of wild-type bacteria. Coating of non-pathogenic E. coli with purified wild-type trehalose dimycolate reduced phagolysosome formation substantially which was not the case with shorter kasA mutant-derived trehalose dimycolate. The mutant was moderately attenuated in macrophages and in a mouse infection model, but was fully cytotoxic.Whereas loss of KasA is lethal in mycobacteria, R. equi kasA mutant multiplication in broth was normal proving that long-chain mycolic acid compounds are not necessarily required for cellular integrity and viability of the bacteria that typically produce them. This study demonstrates a central role of mycolic acid chain length in diversion of trafficking by R. equi. PMID:23078612

  1. Diversion of phagosome trafficking by pathogenic Rhodococcus equi depends on mycolic acid chain length.

    PubMed

    Sydor, Tobias; von Bargen, Kristine; Hsu, Fong-Fu; Huth, Gitta; Holst, Otto; Wohlmann, Jens; Becken, Ulrike; Dykstra, Tobias; Söhl, Kristina; Lindner, Buko; Prescott, John F; Schaible, Ulrich E; Utermöhlen, Olaf; Haas, Albert

    2013-03-01

    Rhodococcus equi is a close relative of Mycobacterium spp. and a facultative intracellular pathogen which arrests phagosome maturation in macrophages before the late endocytic stage. We have screened a transposon mutant library of R. equi for mutants with decreased capability to prevent phagolysosome formation. This screen yielded a mutant in the gene for β-ketoacyl-(acyl carrier protein)-synthase A (KasA), a key enzyme of the long-chain mycolic acid synthesizing FAS-II system. The longest kasA mutant mycolic acid chains were 10 carbon units shorter than those of wild-type bacteria. Coating of non-pathogenic E. coli with purified wild-type trehalose dimycolate reduced phagolysosome formation substantially which was not the case with shorter kasA mutant-derived trehalose dimycolate. The mutant was moderately attenuated in macrophages and in a mouse infection model, but was fully cytotoxic.Whereas loss of KasA is lethal in mycobacteria, R. equi kasA mutant multiplication in broth was normal proving that long-chain mycolic acid compounds are not necessarily required for cellular integrity and viability of the bacteria that typically produce them. This study demonstrates a central role of mycolic acid chain length in diversion of trafficking by R. equi. © 2012 Blackwell Publishing Ltd.

  2. Development of a loop-mediated isothermal amplification method for detecting virulent Rhodococcus equi.

    PubMed

    Kinoshita, Yuta; Niwa, Hidekazu; Higuchi, Tohru; Katayama, Yoshinari

    2016-09-01

    Rhodococcus equi is the most important causative bacterium of severe pneumonia in foals. We report herein the development of a specific loop-mediated isothermal amplification (LAMP) assay, which targets a gene encoding vapA for detecting virulent R. equi The detection limit of the LAMP assay was 10(4) colony forming units (CFU)/mL, which was equal to 10 CFU/reaction. The clinical efficacy of the LAMP assay was compared with those of 2 published PCR-based methods: nested PCR and quantitative real-time (q)PCR. Agreements between bacterial culture, which is the gold standard for detection of R. equi, and each of the 3 molecular tests were measured by calculating a kappa coefficient. The kappa coefficients of the LAMP (0.760), nested PCR (0.583), and qPCR (0.888) indicated substantial agreement, moderate agreement, and almost perfect agreement, respectively. Although the clinical efficacy of LAMP was not the best among the 3 methods tested, LAMP could be more easily introduced into less well-equipped clinics because it does not require special equipment (such as a thermocycler) for gene amplification. Veterinary practitioners could diagnose R. equi pneumonia more quickly by using LAMP and could use the results to select an appropriate initial treatment. © 2016 The Author(s).

  3. Structure of the virulence-associated protein VapD from the intracellular pathogen Rhodococcus equi.

    PubMed

    Whittingham, Jean L; Blagova, Elena V; Finn, Ciaran E; Luo, Haixia; Miranda-CasoLuengo, Raúl; Turkenburg, Johan P; Leech, Andrew P; Walton, Paul H; Murzin, Alexey G; Meijer, Wim G; Wilkinson, Anthony J

    2014-08-01

    Rhodococcus equi is a multi-host pathogen that infects a range of animals as well as immune-compromised humans. Equine and porcine isolates harbour a virulence plasmid encoding a homologous family of virulence-associated proteins associated with the capacity of R. equi to divert the normal processes of endosomal maturation, enabling bacterial survival and proliferation in alveolar macrophages. To provide a basis for probing the function of the Vap proteins in virulence, the crystal structure of VapD was determined. VapD is a monomer as determined by multi-angle laser light scattering. The structure reveals an elliptical, compact eight-stranded β-barrel with a novel strand topology and pseudo-twofold symmetry, suggesting evolution from an ancestral dimer. Surface-associated octyl-β-D-glucoside molecules may provide clues to function. Circular-dichroism spectroscopic analysis suggests that the β-barrel structure is preceded by a natively disordered region at the N-terminus. Sequence comparisons indicate that the core folds of the other plasmid-encoded virulence-associated proteins from R. equi strains are similar to that of VapD. It is further shown that sequences encoding putative R. equi Vap-like proteins occur in diverse bacterial species. Finally, the functional implications of the structure are discussed in the light of the unique structural features of VapD and its partial structural similarity to other β-barrel proteins.

  4. Analysis of equi-intensity curves and NU distribution of EAS

    NASA Astrophysics Data System (ADS)

    Tanahashi, G.

    1985-08-01

    The distribution of the number of muons in extensive air showers (EAS) and the equi-intensity curves of EAS are analyzed on the basis of Monte Carlo simulation of various cosmic ray composition and the interaction models. Problems in the two best combined models are discussed.

  5. Influence of Rhodococcus equi on the respiratory burst of resident alveolar macrophages from horses

    SciTech Connect

    Brumbaugh, G.W.

    1986-01-01

    Rhodococcus equi is the etiologic agent of a devastating pneumonia of sporadic incidence in foals. The purpose of this study was to evaluate the influence of R. equi on the superoxide anion production, measured spectrophotometrically as the reduction of cytochrome C, and hexose monophosphate shunt activity, measured by /sup 14/CO/sub 2/ liberation from /sup 14/C-1-D-glucose, of alveolar macrophages from horses. Alveolar macrophages were harvested from 6 anesthetized, healthy, light-breed, adult horses by bronchoalveolar lavage. Following a randomized complete block design, the suspension of cells was divided into aliquots of 10/sup 6/ viable alveolar macrophages which were exposed to 1, 10 or 100 g. of opsonized R. equi or opsonized zymosan A at 37 C for 2 hours. In this study the respiratory burst of equine alveolar macrophages was only evidenced by the hexose monophosphate shunt activity and superoxide anion was not coincidentally produced. Rhodococcus equi did not adversely affect that response. The insignificant superoxide anion production by the alveolar macrophages suggests that this may not be a significant oxygen metabolite in those cells.

  6. Detection of Babesia caballi and Babesia equi in Dermacentor nuttalli adult ticks.

    PubMed

    Battsetseg, B; Xuan, X; Ikadai, H; Bautista, J L; Byambaa, B; Boldbaatar, D; Battur, B; Battsetseg, G; Batsukh, Z; Igarashi, I; Nagasawa, H; Mikami, T; Fujisaki, K

    2001-04-01

    Ticks play an important role in human and veterinary medicine particularly due to their ability to transmit protozoan pathogens. In this study we have demonstrated that polymerase chain reaction (PCR) and nested PCR methods enabled detection of Babesia caballi and Babesia equi in field isolates of Dermacentor nuttalli adult ticks from Mongolia. Primers specific for 218 bp fragment merozoite antigen 1 (EMA-1) gene of B. equi successfully amplified products from all samples of D. nuttalli adult ticks while primers for the 430 bp fragment product from BC48 gene of B. caballi amplified products from seven of the 54 samples. Using PCR and nested PCR methods we have found mixed infections with B. equi and B. caballi in the tick vector. The amplified DNA fragment from D. nuttalli ticks was inserted into the EcoRV site of pBluescript SK and sequenced. The sequence of the 430 bp fragment was completely identical to the nucleotide sequence of the USDA strain of B. caballi. These results suggest that D. nuttalli may play an important role as a vector of both B. caballi and B. equi and also may be important in maintaining endemicity of equine piroplasmosis in Mongolia.

  7. Microscopic and Molecular Detection of Theileria (Babesia) Equi Infection in Equids of Kurdistan Province, Iran

    PubMed Central

    HABIBI, Gholamreza; ESMAEILNIA, Kasra; HABLOLVARID, Mohammad Hasan; AFSHARI, Asghar; ZAMEN, Mohsen; BOZORGI, Soghra

    2016-01-01

    Background: Equine piroplasmosis (EP) is the cause of persistent tick-borne infection with no symptoms, but the most important problem of EP is due to the persistent carrier state. Carrier animals to Babesia (Theileria) equi (Laveran 1901) and B. caballi (Nuttall, 1910) infestation could be identified by extremely sensitive PCR-based method. The purpose of this study was to identify the causative agents of equine piroplasmosis based on molecular and microscopic assays in equids from Kurdistan Province, Iran. Methods: Thirty one horse and mule blood samples were used with history of living in Kurdistan Province of Iran. The blood specimens were utilized for T. equi and B. caballi DNA identification by PCR and Giemsa stained smears for microscopic observation. Results: The results clearly showed the presence of B. (Theileria) equi DNA in 30 of 31 blood samples (96.77%), but the microscopic examination revealed the 3 of 31 positive Babesia like organisms in the red blood cells (9.67%). Conclusion: The obtained results demonstrated the presence of hidden B. (Theileria) equi infection in horses with previous habitance in Kurdistan Province of Iran. The carrier animals became a main source of infection and can transmit the disease. Therefore, hidden infection might be considered as a health threatening and limiting factor in animals used in therapeutic antisera research and production centers. PMID:27095973

  8. Molecular detection and prevalence of Theileria equi and Babesia caballi in horses of central Balkan.

    PubMed

    Davitkov, Darko; Vucicevic, Milos; Stevanovic, Jevrosima; Krstic, Vanja; Slijepcevic, Dajana; Glavinic, Uros; Stanimirovic, Zoran

    2016-03-01

    Equine piroplasmosis is significant tick-borne disease with wide distribution. The prevalence of equine piroplasmosis in Serbia, Montenegro and Bosnia and Herzegovina is unknown. In aim to obtain a first insight into the prevalence we performed molecular epidemiological study which included 142 horses, on seven locations in these three countries. We first performed PCR for the detection of a 450bp long section of the 18S rRNA of piroplasma-specific region. For all positive samples we have done multiplex PCR for the species detection. Species determination was further confirmed by sequencing PCR products of 10 randomly selected Theileria equi and all Babesia caballi samples. The overall prevalence rates in analysed region for T. equi and B. caballi were 22.5% and 2.1%, respectively. Possible risk factors (such as location, age, sex and activity) associated with PCR positivity were evaluated. Marked differences were found in prevalence between geographic areas. There was no significant association between positivity and age group. T. equi was more prevalent in females and farming horses. This is the first report on the molecular survey of T. equi and B. caballi in central Balkan. Further prevalence studies on definitive host and vectors in this region are necessary.

  9. Rhodococcus equi infection in HIV-positive subjects: a retrospective analysis of 24 cases.

    PubMed

    Arlotti, M; Zoboli, G; Moscatelli, G L; Magnani, G; Maserati, R; Borghi, V; Andreoni, M; Libanore, M; Bonazzi, L; Piscina, A; Ciammarughi, R

    1996-01-01

    Rhodococcus equi causes a rare infection in immunocompromised hosts. We describe 24 cases of infection in patients with AIDS-related complex (ARC)/acquired immunodeficiency syndrome (AIDS). Pneumonia was always the first manifestation of R. equi infection, but extrapulmonary involvement was also observed. The main sources of bacteria were sputum, bronchial washings and blood. The strains isolated were mainly susceptible to erythromycin, vancomycin, teicoplanin, rifampicin, imipenem and aminoglycosides. Initial treatment should involve an intravenously administered antibiotic combination therapy including imipenem or vancomycin or teicoplanin, followed by orally administered maintenance combination therapy. Drug combinations should be investigated for serum bactericidal activity in vitro. Surgery does not increase survival time and should only be performed in cases that do not respond to antibiotic treatment. Presumptive risks of infection (contact with horses or farm dust, or cohabiting with people affected by R. equi infection) were present in more than 50% of patients. This finding, and the frequency of bacteria in the sputum, are not sufficient proof of transmission between humans, but do suggest the need for respiratory isolation of patients affected by R. equi pneumonia.

  10. Assessing the Genotypic Differences between Strains of Corynebacterium pseudotuberculosis biovar equi through Comparative Genomics

    PubMed Central

    Ramos, Rommel T. J.; Veras, Adonney A. O.; Pinheiro, Kenny C.; Benevides, Leandro J.; Edman, Judy M.; Spier, Sharon J.; Azevedo, Vasco; Silva, Artur

    2017-01-01

    Seven genomes of Corynebacterium pseudotuberculosis biovar equi were sequenced on the Ion Torrent PGM platform, generating high-quality scaffolds over 2.35 Mbp. This bacterium is the causative agent of disease known as “pigeon fever” which commonly affects horses worldwide. The pangenome of biovar equi was calculated and two phylogenomic approaches were used to identify clustering patterns within Corynebacterium genus. Furthermore, other comparative analyses were performed including the prediction of genomic islands and prophages, and SNP-based phylogeny. In the phylogenomic tree, C. pseudotuberculosis was divided into two distinct clades, one formed by nitrate non-reducing species (biovar ovis) and another formed by nitrate-reducing species (biovar equi). In the latter group, the strains isolated from California were more related to each other, while the strains CIP 52.97 and 1/06-A formed the outermost clade of the biovar equi. A total of 1,355 core genes were identified, corresponding to 42.5% of the pangenome. This pangenome has one of the smallest core genomes described in the literature, suggesting a high genetic variability of biovar equi of C. pseudotuberculosis. The analysis of the similarity between the resistance islands identified a higher proximity between the strains that caused more severe infectious conditions (infection in the internal organs). Pathogenicity islands were largely conserved between strains. Several genes that modulate the pathogenicity of C. pseudotuberculosis were described including peptidases, recombination enzymes, micoside synthesis enzymes, bacteriocins with antimicrobial activity and several others. Finally, no genotypic differences were observed between the strains that caused the three different types of infection (external abscess formation, infection with abscess formation in the internal organs, and ulcerative lymphangitis). Instead, it was noted that there is a higher phenetic correlation between strains isolated at

  11. Minimum inhibitory concentrations of erythromycin and rifampin for Rhodococcus equi during the years 2007-2014.

    PubMed

    Fenton, Caitriona S; Buckley, Thomas C

    2015-01-01

    Rhodococcus equi is a gram positive, intracellular pathogen of foals worldwide. The aim of this study was to determine whether there was an increasing resistance occurring in Rhodococcus equi towards the antibiotics rifampin and erythromycin over a seven year period. The investigation was carried out with the use of E test strips (epsilometers) for rifampin and erythromycin in order to determine the Minimum Inhibitory Concentrations (MIC) values of Rhodococcus equi to these antibiotics. The main results of this study found that the mean MICs were higher for erythromycin than for rifampin for every year analysed apart from 2008. The results highlight that 75 % (6/8) of the mean MICs for erythromycin were above the threshold of susceptibility of 0.5 μg/ml and one of the yearly mean MICs for rifampin (2008) was above the level of ≤ 1 μg/ml. Two soil samples analysed had high MIC values of 2 μg/ml and 3 μg/ml for rifampin and erythromycin respectively. These samples can be said to have acquired resistance as they are above 1 μg/ml. The significance of these findings is that R. equi is already a problematic pathogen to treat and if the bacteria keeps gaining resistance to these antibiotics at rate that has been shown over the last decade, then a new form of treatment will have to be introduced. Further research into the genomics of Rhodococcus equi will, in time, shed more light on possible alternatives such as vaccines or new, more effective antimicrobials.

  12. Structure of the virulence-associated protein VapD from the intracellular pathogen Rhodococcus equi

    SciTech Connect

    Whittingham, Jean L.; Blagova, Elena V.; Finn, Ciaran E.; Luo, Haixia; Miranda-CasoLuengo, Raúl; Turkenburg, Johan P.; Leech, Andrew P.; Walton, Paul H.; Murzin, Alexey G.; Meijer, Wim G.; Wilkinson, Anthony J.

    2014-08-01

    VapD is one of a set of highly homologous virulence-associated proteins from the multi-host pathogen Rhodococcus equi. The crystal structure reveals an eight-stranded β-barrel with a novel fold and a glycine rich ‘bald’ surface. Rhodococcus equi is a multi-host pathogen that infects a range of animals as well as immune-compromised humans. Equine and porcine isolates harbour a virulence plasmid encoding a homologous family of virulence-associated proteins associated with the capacity of R. equi to divert the normal processes of endosomal maturation, enabling bacterial survival and proliferation in alveolar macrophages. To provide a basis for probing the function of the Vap proteins in virulence, the crystal structure of VapD was determined. VapD is a monomer as determined by multi-angle laser light scattering. The structure reveals an elliptical, compact eight-stranded β-barrel with a novel strand topology and pseudo-twofold symmetry, suggesting evolution from an ancestral dimer. Surface-associated octyl-β-d-glucoside molecules may provide clues to function. Circular-dichroism spectroscopic analysis suggests that the β-barrel structure is preceded by a natively disordered region at the N-terminus. Sequence comparisons indicate that the core folds of the other plasmid-encoded virulence-associated proteins from R. equi strains are similar to that of VapD. It is further shown that sequences encoding putative R. equi Vap-like proteins occur in diverse bacterial species. Finally, the functional implications of the structure are discussed in the light of the unique structural features of VapD and its partial structural similarity to other β-barrel proteins.

  13. Assessing the Genotypic Differences between Strains of Corynebacterium pseudotuberculosis biovar equi through Comparative Genomics.

    PubMed

    Baraúna, Rafael A; Ramos, Rommel T J; Veras, Adonney A O; Pinheiro, Kenny C; Benevides, Leandro J; Viana, Marcus V C; Guimarães, Luís C; Edman, Judy M; Spier, Sharon J; Azevedo, Vasco; Silva, Artur

    2017-01-01

    Seven genomes of Corynebacterium pseudotuberculosis biovar equi were sequenced on the Ion Torrent PGM platform, generating high-quality scaffolds over 2.35 Mbp. This bacterium is the causative agent of disease known as "pigeon fever" which commonly affects horses worldwide. The pangenome of biovar equi was calculated and two phylogenomic approaches were used to identify clustering patterns within Corynebacterium genus. Furthermore, other comparative analyses were performed including the prediction of genomic islands and prophages, and SNP-based phylogeny. In the phylogenomic tree, C. pseudotuberculosis was divided into two distinct clades, one formed by nitrate non-reducing species (biovar ovis) and another formed by nitrate-reducing species (biovar equi). In the latter group, the strains isolated from California were more related to each other, while the strains CIP 52.97 and 1/06-A formed the outermost clade of the biovar equi. A total of 1,355 core genes were identified, corresponding to 42.5% of the pangenome. This pangenome has one of the smallest core genomes described in the literature, suggesting a high genetic variability of biovar equi of C. pseudotuberculosis. The analysis of the similarity between the resistance islands identified a higher proximity between the strains that caused more severe infectious conditions (infection in the internal organs). Pathogenicity islands were largely conserved between strains. Several genes that modulate the pathogenicity of C. pseudotuberculosis were described including peptidases, recombination enzymes, micoside synthesis enzymes, bacteriocins with antimicrobial activity and several others. Finally, no genotypic differences were observed between the strains that caused the three different types of infection (external abscess formation, infection with abscess formation in the internal organs, and ulcerative lymphangitis). Instead, it was noted that there is a higher phenetic correlation between strains isolated at

  14. Fine structure of Babesia equi Laveran, 1901 within lymphocytes and erythrocytes of horses: an in vivo and in vitro study.

    PubMed

    Moltmann, U G; Mehlhorn, H; Schein, E; Rehbein, G; Voigt, W P; Zweygarth, E

    1983-02-01

    The development of Babesia equi (Piroplasmia) in the vertebrate host was studied by electron microscopy. The tick-borne sporozoites initiated an exoerythrocytic schizogony in lymphocytes in vivo and in vitro. In lymphocyte cultures the schizonts of B. equi lay as fissured bodies directly within the host cell forming numerous cytomeres. These cytomeres were bordered by a single membrane and contained polymorphous nuclei. Merozoite differentiation was initiated at several places by protrusions appearing at the periphery of the cytomeres. The cytoplasm of the schizont merged progressively into the young merozoites. The mature merozoites were pear-shaped (2.0 X 1.5 microns), bounded by a pellicle and had an apical complex without conoid. Numerous merozoites were observed entering erythrocytes in vitro. In vivo, lymphocytes containing B. equi schizonts were seen in capillaries within lymph nodes from the 12th day onwards after attachment of infected ticks. At the same time some erythrocytes were found that contained B. equi merozoites. Development of B. equi in the vertebrate host shows many similarities to the corresponding development of Theileria parasites. This makes a new discussion of the taxonomic position of B. equi necessary.

  15. An outbreak of Streptococcus equi subspecies zooepidemicus associated with consumption of fresh goat cheese

    PubMed Central

    Kuusi, Markku; Lahti, Elina; Virolainen, Anni; Hatakka, Maija; Vuento, Risto; Rantala, Leila; Vuopio-Varkila, Jaana; Seuna, Eija; Karppelin, Matti; Hakkinen, Marjaana; Takkinen, Johanna; Gindonis, Veera; Siponen, Kyosti; Huotari, Kaisa

    2006-01-01

    Background Streptococcus equi subspecies zooepidemicus is a rare infection in humans associated with contact with horses or consumption of unpasteurized milk products. On October 23, 2003, the National Public Health Institute was alerted that within one week three persons had been admitted to Tampere University Central Hospital (TaYS) because of S. equi subsp. zooepidemicus septicaemia. All had consumed fresh goat cheese produced in a small-scale dairy located on a farm. We conducted an investigation to determine the source and the extent of the outbreak. Methods Cases were identified from the National Infectious Disease Register. Cases were persons with S. equi subsp. zooepidemicus isolated from a normally sterile site who had illness onset 15.9-31.10.2003. All cases were telephone interviewed by using a standard questionnaire and clinical information was extracted from patient charts. Environmental and food specimens included throat swabs from two persons working in the dairy, milk from goats and raw milk tank, cheeses made of unpasteurized milk, vaginal samples of goats, and borehole well water. The isolates were characterized by ribotyping and pulsed-field gel electrophoresis (PFGE). Results Seven persons met the case definition; six had septicaemia and one had purulent arthritis. Five were women; the median age was 70 years (range 54–93). None of the cases were immunocompromized and none died. Six cases were identified in TaYS, and one in another university hospital in southern Finland. All had eaten goat cheese produced on the implicated farm. S. equi subsp. zooepidemicus was isolated from throat swabs, fresh goat cheese, milk tank, and vaginal samples of one goat. All human and environmental strains were indistinguishable by ribotyping and PFGE. Conclusion The outbreak was caused by goat cheese produced from unpasteurized milk. Outbreaks caused by S. equi subsp. zooepidemicus may not be detected if streptococcal strains are only typed to the group level. S

  16. Evaluation of Veterinary-Specific Interpretive Criteria for Susceptibility Testing of Streptococcus equi Subspecies with Trimethoprim-Sulfamethoxazole and Trimethoprim-Sulfadiazine

    PubMed Central

    Kanellos, Theo; Guardabassi, Luca; Boucher, Joseph

    2016-01-01

    ABSTRACT Antimicrobial susceptibility test results for trimethoprim-sulfadiazine with Streptococcus equi subspecies are interpreted based on human data for trimethoprim-sulfamethoxazole. The veterinary-specific data generated in this study support a single breakpoint for testing trimethoprim-sulfamethoxazole and/or trimethoprim-sulfadiazine with S. equi. This study indicates trimethoprim-sulfamethoxazole as an acceptable surrogate for trimethoprim-sulfadiazine with S. equi. PMID:27847375

  17. Evaluation of Veterinary-Specific Interpretive Criteria for Susceptibility Testing of Streptococcus equi Subspecies with Trimethoprim-Sulfamethoxazole and Trimethoprim-Sulfadiazine.

    PubMed

    Sadaka, Carmen; Kanellos, Theo; Guardabassi, Luca; Boucher, Joseph; Watts, Jeffrey L

    2017-01-01

    Antimicrobial susceptibility test results for trimethoprim-sulfadiazine with Streptococcus equi subspecies are interpreted based on human data for trimethoprim-sulfamethoxazole. The veterinary-specific data generated in this study support a single breakpoint for testing trimethoprim-sulfamethoxazole and/or trimethoprim-sulfadiazine with S. equi This study indicates trimethoprim-sulfamethoxazole as an acceptable surrogate for trimethoprim-sulfadiazine with S. equi. Copyright © 2016 Sadaka et al.

  18. Prevalence and genetic diversity of Rhodococcus equi in wild boars (Sus scrofa), roe deer (Capreolus capreolus) and red deer (Cervus elaphus) in Poland.

    PubMed

    Witkowski, Lucjan; Rzewuska, Magdalena; Cisek, Agata Anna; Chrobak-Chmiel, Dorota; Kizerwetter-Świda, Magdalena; Czopowicz, Michał; Welz, Mirosław; Kita, Jerzy

    2015-05-22

    Rhodococcus equi is now considered an emerging zoonotic pathogen. Sources and routes of human infection remain unclear but foodborne transmission seems to be the most probable way. Strains of pig or bovine type are most often isolated from human cases and moreover R. equi is present in submaxillary lymph nodes of apparently healthy pigs and wild boars intended for human consumption. The aim of this study was to estimate the prevalence of R. equi in submaxillary lymph nodes in wild boars, roe deer and red deer. Samples were collected from 936 animals and 27 R. equi strains were isolated, from 5.1 % of wild boars (23/452), 0.7 % of red deer (2/272) and 0.9 % of roe deer (2/212). Genetic diversity of all 27 isolates was studied using VspI-PFGE method, resulting in the detection of 25 PFGE patterns and four PFGE clusters. PFGE patterns of the isolates were compared with virulence plasmid types and no concordance was observed. R. equi was present in wild animal tissues and consumption of the game may be a potential source of R. equi infection for humans. To the authors' best knowledge, this is the first epidemiological report of R. equi prevalence in tissues of roe deer and red deer. However, risk associated with wild ruminant consumption seems marginal. Investigation of R. equi transmission between animals and humans based exclusively on types of virulence plasmids seems to be insufficient to identify sources of R. equi infection for people.

  19. Antibiotic failure in a renal transplant patient with Rhodococcus equi infection: an indication for surgical lobectomy.

    PubMed

    Ursales, A; Klein, J A; Beal, S G; Koch, M; Clement-Kruzel, S; Melton, L B; Spak, C W

    2014-12-01

    Rhodococcus equi is an animal pathogen that causes infrequent but challenging infections in immunocompromised individuals, few of which have been described in solid organ transplant recipients. Common clinical presentations include indolent cough, fever, and dyspnea, with necrotizing pneumonia and cavitation. We report a case of a dense right upper lung pneumonia with resultant R. equi bacteremia in a renal transplant recipient. Our patient initially responded to antibiotic treatment with resolution of bacteremia and clinical recovery, followed by interval progression in her right upper lobe consolidation on follow-up computed tomography scans. She underwent lobectomy for definitive therapy with resolution of symptoms. Lobectomy can be utilized in isolated infection after antibiotic failure with excellent clinical outcomes. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  20. Protection of foals against experimental Rhodococcus equi pneumonia by oral immunization.

    PubMed Central

    Chirino-Trejo, J M; Prescott, J F; Yager, J A

    1987-01-01

    Two groups of three one to three week old foals were immunized orally on four occasions over five weeks with two strains of Rhodococcus equi, a clinical isolate from a pneumonic foal and a laboratory passaged Congo red negative variant of this strain. Three nonimmunized foals of similar age acted as controls. Three weeks after the last immunization, all foals were challenged on five occasions over seven days by aerosol infection with about 10(10) of the pneumonic foal isolate on each occasion. Control foals became seriously ill and were euthanized. Immunization with either strain protected foals equally against the challenge, and resulted in rapid lung clearance. Oral immunization can thus protect foals against severe challenge with R. equi. The proteins associated with Congo red colony staining appear not to be involved in protective immunity. PMID:3453264

  1. Seroepidemiological survey of Rhodococcus equi infection in asymptomatic horses and donkeys from southeast Turkey.

    PubMed

    Tel, O Y; Arserim, N B; Keskin, O

    2011-12-01

    In order to assess the level of Rhodococcus equi infection in southeast Turkey, 679 sera from healthy foals and adult horses and 78 sera from donkeys were tested by indirect ELISA using a R. equi reference strain (ATCC 33701) as antigen. Eighty (11.7%) sera from horses and 9 (11.5%) sera from donkeys with titres >0.85 were positive. The prevalence of seropositive horses in Sanliurfa Province was higher than in Diyarbakir Province; 56 (13.9%) horses in Sanliurfa Province and 24 (8.7%) horses in Diyarbakir Province were defined as seropositive. In Sanliurfa Province 14.5% of female (n=343) and 10.1% of male (n = 59) horses tested were defined as seropositive, while in Diyarbakir Province more males (11.4%, n=114) were seropositive than females (6.7%, n=163). Horses 1 to 5 years of age were found to have the highest seropositivity rate in both provinces. A total of 78 sera from donkeys were investigated in Sanliurfa Province, of which 9 (11.5%) were positive by ELISA. Among the 9 positive sera, 6 (12.8%) were from donkeys 1-5 years old and 3 (13.6%) were from donkeys >5 years of age. No positive sera were found in donkeys less than 1 year old. Five (12.5%) sera of females and 4 (10.5%) sera of males tested were positive. These results indicate the existence of R. equi in the horse populations in Sanliurfa and Diyarbakir Provinces. Similar infection rates were found for donkeys in Sanliurfa. This suggests the importance of serological surveys to diagnose R. equi infection in the region and to prevent the zoonotic risk.

  2. Systemic and respiratory oxidative stress in the pathogenesis and diagnosis of Rhodococcus equi pneumonia.

    PubMed

    Crowley, J; Po, E; Celi, P; Muscatello, G

    2013-12-01

    Oxidative stress (OS) is most simply defined as an imbalance between oxidants and antioxidants. Oxidative stress has been suggested to play roles in various equine respiratory diseases and the significance of OS in the pathogenesis of Rhodococcus equi pneumonia is unknown. To measure and relate biomarkers of OS to lesions consistent with R. equi pneumonia. Case-control study. Various OS biomarkers were measured from blood and exhaled breath condensate (EBC) samples collected from 26 foals between 1 and 2 months of age (n = 12 cases and n = 14 controls) on 2 Thoroughbred farms endemically affected by R. equi pneumonia. Foals were defined as cases (positive) or controls (negative) based on ultrasonographic evidence of pulmonary abscessation (>15 mm in diameter). Haematology and biochemistry testing was also performed on blood samples collected from the foals. Comparison of biomarkers and key haematological and biochemical markers of inflammation between the groups was performed using 2 sample t tests. Derivatives of reactive oxygen metabolites (d-ROMs) were significantly greater in case foals than in control foals (P = 0.027) and the oxidative stress index (OSI) was higher in case foals (P = 0.014). Hydrogen peroxide (H2 O2 ) concentrations in EBC were significantly greater in case foals than in control foals (P = 0.002). Meanwhile, there were no significant differences in traditional measures of inflammation between the 2 groups. Measuring OS in both blood and EBC provided useful information in the early diagnosis of R. equi pneumonia. © 2013 EVJ Ltd.

  3. IcgA is a virulence factor of Rhodococcus equi that modulates intracellular growth.

    PubMed

    Wang, Xiaoguang; Coulson, Garry B; Miranda-Casoluengo, Aleksandra A; Miranda-Casoluengo, Raúl; Hondalus, Mary K; Meijer, Wim G

    2014-05-01

    Virulence of the intracellular pathogen Rhodococcus equi depends on a 21.3-kb pathogenicity island located on a conjugative plasmid. To date, the only nonregulatory pathogenicity island-encoded virulence factor identified is the cell envelope-associated VapA protein. Although the pathogenicity islands from porcine and equine R. equi isolates have undergone major rearrangements, the virR operon (virR-icgA-vapH-orf7-virS) is highly conserved in both, suggesting these genes play an important role in pathogenicity. VirR and VirS are transcriptional regulators controlling expression of pathogenicity island genes, including vapA. Here, we show that while vapH and orf7 are dispensable for intracellular growth of R. equi, deletion of icgA, formerly known as orf5, encoding a major facilitator superfamily transport protein, elicited an enhanced growth phenotype in macrophages and a significant reduction in macrophage viability, while extracellular growth in broth remained unaffected. Transcription of virS, located downstream of icgA, and vapA was not affected by the icgA deletion during growth in broth or in macrophages, showing that the enhanced growth phenotype caused by deletion of icgA was not mediated through abnormal transcription of these genes. Transcription of icgA increased 6-fold within 2 h following infection of macrophages and remained significantly higher 48 h postinfection compared to levels at the start of the infection. The major facilitator superfamily transport protein IcgA is the first factor identified in R. equi that negatively affects intracellular replication. Aside from VapA, it is only the second pathogenicity island-encoded structural protein shown to play a direct role in intracellular growth of this pathogenic actinomycete.

  4. IcgA Is a Virulence Factor of Rhodococcus equi That Modulates Intracellular Growth

    PubMed Central

    Wang, Xiaoguang; Coulson, Garry B.; Miranda-CasoLuengo, Aleksandra A.; Miranda-CasoLuengo, Raúl; Hondalus, Mary K.

    2014-01-01

    Virulence of the intracellular pathogen Rhodococcus equi depends on a 21.3-kb pathogenicity island located on a conjugative plasmid. To date, the only nonregulatory pathogenicity island-encoded virulence factor identified is the cell envelope-associated VapA protein. Although the pathogenicity islands from porcine and equine R. equi isolates have undergone major rearrangements, the virR operon (virR-icgA-vapH-orf7-virS) is highly conserved in both, suggesting these genes play an important role in pathogenicity. VirR and VirS are transcriptional regulators controlling expression of pathogenicity island genes, including vapA. Here, we show that while vapH and orf7 are dispensable for intracellular growth of R. equi, deletion of icgA, formerly known as orf5, encoding a major facilitator superfamily transport protein, elicited an enhanced growth phenotype in macrophages and a significant reduction in macrophage viability, while extracellular growth in broth remained unaffected. Transcription of virS, located downstream of icgA, and vapA was not affected by the icgA deletion during growth in broth or in macrophages, showing that the enhanced growth phenotype caused by deletion of icgA was not mediated through abnormal transcription of these genes. Transcription of icgA increased 6-fold within 2 h following infection of macrophages and remained significantly higher 48 h postinfection compared to levels at the start of the infection. The major facilitator superfamily transport protein IcgA is the first factor identified in R. equi that negatively affects intracellular replication. Aside from VapA, it is only the second pathogenicity island-encoded structural protein shown to play a direct role in intracellular growth of this pathogenic actinomycete. PMID:24549327

  5. Mutant prevention concentration and mutant selection window for 10 antimicrobial agents against Rhodococcus equi.

    PubMed

    Berghaus, Londa J; Giguère, Steeve; Guldbech, Kristen

    2013-10-25

    The objectives of this study were to determine the mutant prevention concentration (MPC), time above the MPC and mutant selection window for 10 antimicrobial agents against Rhodococcus equi and to determine if the combination of a macrolide with rifampin would decrease emergence of resistant mutants. Antimicrobial agents investigated (erythromycin, clarithromycin, azithromycin, rifampin, amikacin, gentamicin, enrofloxacin, vancomycin, imipenem, and doxycycline) were selected based on in vitro activity and frequency of use in foals or people infected with R. equi. Each antimicrobial agent or combination of agents was evaluated against four virulent strains of R. equi. MPC were determined using an agar plate assay. Pharmacodynamic parameters were calculated using published plasma and pulmonary pharmacokinetic variables. There was a significant (P<0.001) effect of the type of antimicrobial agent on the MPC. The MPC of clarithromycin (1.0 μg/ml) was significantly lower and the MPC of rifampin and amikacin (512 and 384 μg/ml, respectively) were significantly higher than that of all other antimicrobial agents tested. Combining erythromycin, clarithromycin, or azithromycin with rifampin resulted in a significant (P≤0.005) decrease in MPC and MPC/MIC ratio. When MIC and MPC were combined with pharmacokinetic variables, only gentamicin and vancomycin were predicted to achieve plasma concentrations above the MPC for any given periods of time. Only clarithromycin and the combination clarithromycin-rifampin were predicted to achieve concentrations in bronchoalveolar cells and pulmonary epithelial lining fluid above the MPC for the entire dosing interval. In conclusion, the combination of a macrolide with rifampin considerably decreases the emergence of resistant mutants of R. equi. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. MICs of 32 antimicrobial agents for Rhodococcus equi isolates of animal origin.

    PubMed

    Riesenberg, Anne; Feßler, Andrea T; Erol, Erdal; Prenger-Berninghoff, Ellen; Stamm, Ivonne; Böse, Reinhard; Heusinger, Anton; Klarmann, Dieter; Werckenthin, Christiane; Schwarz, Stefan

    2014-04-01

    The aim of this study was to determine the MICs of 32 antimicrobial agents for 200 isolates of Rhodococcus equi of animal origin by applying a recently described broth microdilution protocol, and to investigate isolates with distinctly elevated rifampicin MICs for the genetic basis of rifampicin resistance. The study included 200 R. equi isolates, including 160 isolates from horses and 40 isolates from other animal sources, from the USA and Europe. MIC testing of 32 antimicrobial agents or combinations thereof followed a recently published protocol. A novel PCR protocol for the joint amplification of the three rpoB regions in which rifampicin resistance-mediating mutations have been reported was applied to isolates with elevated rifampicin MICs. The amplicons were sequenced and screened for mutations. Susceptibility testing revealed a rather uniform distribution of MICs for most of the antimicrobial agents tested. The lowest MICs were seen for clarithromycin, rifampicin and imipenem. Six isolates (3%) exhibited distinctly higher MICs of rifampicin than the remaining 194 isolates. In five of these six isolates, single bp exchanges, which resulted in the amino acid exchanges Gln513Leu, Asp516Val, His526Asp or Ser531Leu, were detected in the rifampicin resistance-determining region 1 of the rpoB gene, with Gln513Leu representing a novel substitution for R. equi. This study shows the MIC distribution of 32 antimicrobial agents for a large collection of R. equi isolates of animal origin from two continents. Isolates that exhibited distinctly elevated MICs of rifampicin were only rarely detected.

  7. Prophage lysin Ply30 protects mice from Streptococcus suis and Streptococcus equi subsp. zooepidemicus infections.

    PubMed

    Tang, Fang; Li, Dezhi; Wang, Haojin; Ma, Zhe; Lu, Chengping; Dai, Jianjun

    2015-11-01

    Streptococcus suis and Streptococcus equi subsp. zooepidemicus are capable of infecting humans and various animals, causing significant problems for the worldwide swine industry. As antibiotic resistance has increased, lysosomal enzymes encoded by phages have shown potential for use against pathogenic bacteria. In this study, a novel bacteriophage lysin, Ply30, encoded by the S. suis prophage phi30c, was recombinantly expressed and purified. Ply30 showed high bacteriolysis activity on S. suis and S. equi subsp. zooepidemicus in vitro. The ratio of the optical density at 600 nm (OD600) with treatment versus the OD600 with no treatment for most tested S. suis and S. equi subsp. zooepidemicus strains decreased from 1 to <0.3 and <0.5, respectively, within 1 h. The results of plate viability assays showed that treated bacteria suffered a 1- to 2-log decrease in CFU within 1 h. The optimal concentration of Ply30 was 50 μg/ml, and the optimal pH was 7. Moreover, Ply30 maintained high activity over a wide pH range (pH 6 to 10). The MICs of Ply30 against Streptococcus strains ranged from 16 to 512 μg/ml. In vivo, a 2-mg dose of Ply30 protected 90% (9/10 mice) of mice from infection with S. equi subsp. zooepidemicus and 80% (8/10 mice) of mice from infection with S. suis. Seven days after lysin Ply30 treatment, bacterial loads were significantly decreased in all tested organs and blood compared with those at 1 h postinfection without Ply30 treatment. Ply30 showed in vitro and in vivo antimicrobial efficiency and protected mice against two kinds of bacterial infections, indicating that Ply30 may be an effective therapeutic against streptococci. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. Disseminated rhodococcus equi infection in HIV infection despite highly active antiretroviral therapy

    PubMed Central

    2011-01-01

    Background Rhodococcus equi (R.equi) is an acid fast, GRAM + coccobacillus, which is widespread in the soil and causes pulmonary and extrapulmonary infections in immunocompromised people. In the context of HIV infection, R.equi infection (rhodococcosis) is regarded as an opportunistic disease, and its outcome is influenced by highly active antiretroviral therapy (HAART). Case presentation We report two cases of HIV-related rhodococcosis that disseminated despite suppressive HAART and anti-rhodococcal treatment; in both cases there was no immunological recovery, with CD4+ cells count below 200/μL. In the first case, pulmonary rhodococcosis presented 6 months after initiation of HAART, and was followed by an extracerebral intracranial and a cerebral rhodococcal abscess 1 and 8 months, respectively, after onset of pulmonary infection. The second case was characterized by a protracted course with spread of infection to various organs, including subcutaneous tissue, skin, colon and other intra-abdominal tissues, and central nervous system; the spread started 4 years after clinical resolution of a first pulmonary manifestation and progressed over a period of 2 years. Conclusions Our report highlights the importance of an effective immune recovery, despite fully suppressive HAART, along with anti-rhodococcal therapy, in order to clear rhodococcal infection. PMID:22168333

  9. The Rhodococcus equi virulence protein VapA disrupts endolysosome function and stimulates lysosome biogenesis.

    PubMed

    Rofe, Adam P; Davis, Luther J; Whittingham, Jean L; Latimer-Bowman, Elizabeth C; Wilkinson, Anthony J; Pryor, Paul R

    2017-04-01

    Rhodococcus equi (R. equi) is an important pulmonary pathogen in foals that often leads to the death of the horse. The bacterium harbors a virulence plasmid that encodes numerous virulence-associated proteins (Vaps) including VapA that is essential for intracellular survival inside macrophages. However, little is known about the precise function of VapA. Here, we demonstrate that VapA causes perturbation to late endocytic organelles with swollen endolysosome organelles having reduced Cathepsin B activity and an accumulation of LBPA, LC3 and Rab7. The data are indicative of a loss of endolysosomal function, which leads cells to upregulate lysosome biogenesis to compensate for the loss of functional endolysosomes. Although there is a high degree of homology of the core region of VapA to other Vap proteins, only the highly conserved core region of VapA, and not VapD of VapG, gives the observed effects on endolysosomes. This is the first demonstration of how VapA works and implies that VapA aids R. equi survival by reducing the impact of lysosomes on phagocytosed bacteria. © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  10. A strain device imposing dynamic and uniform equi-biaxial strain to cultured cells.

    PubMed

    Sotoudeh, M; Jalali, S; Usami, S; Shyy, J Y; Chien, S

    1998-01-01

    The objective of this study is to design a new apparatus to allow the control of the magnitude and frequency of dynamic stretch applied uniformly to cells cultured on a silicon elastic membrane. The apparatus is designed to produce equi-biaxial dynamic stretches with area changes ranging from 0% to 55% and frequencies ranging from 0 to 2 Hz. Homogeneous finite strain analysis using triangles of markers was performed to compute the symmetric two-dimensional Lagrangian strain tensor on the membrane. Measurements of strain in both static and dynamic conditions showed that the shear component of the strain tensor (Erc) was near zero, and that there was no significant difference between radial (Err) and circumferential (Ecc) components, indicating the attainment of equi-biaxial strain. Bovine aortic endothelial cells were transiently transfected with a chimeric construct in which the luciferase reporter is driven by TPA-responsive elements (TRE). The transfected cells cultured on the membrane were stretched. The luciferase activity increased significantly only when the cells were stretched by 15% or more in area. Cells in different locations of the membrane showed similar induction of luciferase activities, confirming that strain is uniform and equi-biaxial across the membrane.

  11. A novel ROM compression architecture for DDFS utilizing the parabolic approximation of equi-section division.

    PubMed

    Jeng, Shiann-Shiun; Lin, Hsing-Chen; Lin, Chi-Huei

    2012-12-01

    In this paper, we propose the parabolic approximation of equi-section division (PAESD) utilizing the symmetry property and amplitude approximation of a sinusoidal waveform to design a direct digital frequency synthesizer (DDFS). The sinusoidal phase of a one-quarter period is divided into equi-sections. The proposed method utilizes the curvature equivalence to derive each parabolic curve function, and then the value of the error function between each parabolic curve function and the sinusoidal function is stored in an error-compensation ROM to reconstruct the real sinusoidal waveform. The upper/lower bound of the maximum error value stored in the error-compensation ROM is derived to determine the minimum required memory word length relative to the number of bits of the equi-sections. Thus, the minimum size of the total ROMs of the DDFS using the PAESD without error-compensation ROM is compressed to 544 bits; the total compression ratio, compared with the minimum size of the total ROMs of the DDFS using the basic look-up table (LUT), is approximately 843:1, achieved by consuming additional circuits [71 adaptive look-up tables (ALUTs), 3 digital signal processor (DSP) block 9-bit elements]. Consequently, the results show that the proposed ROM compression method can effectively achieve a better compression ratio than the state-of-the-art solutions without affecting the spectrum performance of an average spurious-free dynamic range (SFDR) of -85 dBc.

  12. Seroprevalence of Theileria equi and Babesia caballi in horses in Spain

    PubMed Central

    Montes Cortés, Maria Guadalupe; Fernández-García, José Luis; Habela Martínez-Estéllez, Miguel Ángel

    2017-01-01

    Equine piroplasmoses are enzootic parasitic diseases distributed worldwide with high incidence in tropical and subtropical regions. In Spain, there is insufficient epidemiological data about equine piroplasmoses. The main aim of the present study was therefore to estimate the prevalence of Theileria equi and Babesia caballi in five regions and obtain information about the risk factors. This study was conducted in the central and south-western regions of Spain, using indirect fluorescence antibody testing (IFAT) in 3,100 sera samples from apparently healthy horses of different ages, breeds, coat colours, genders and geographical locations. The overall seroprevalence was 52%, consisting of 44% seropositive for T. equi and 21% for B. caballi. There was a significant association between age (p < 0.0001), breed (p < 0.004), geographical location (p < 0.0001) and the seroprevalence, but neither the coat colour nor the gender was significantly associated with prevalence. In addition, it was proved that most of the geographic areas showed a moderate to high prevalence. The statistical κ value was used to compare the results obtained by the IFAT and the competitive enzyme-linked immunosorbent assay (cELISA) utilised to test some samples (n = 108) and showed a higher concordance for T. equi (κ = 0.68) than for B. caballi (κ = 0.22). Consequently, this revealed the importance of developing an appropriate technique to detect each haemoparasite. PMID:28497743

  13. Infestation of Werneckiella equi on Icelandic horses, characteristics of predilection sites and lice dermatitis.

    PubMed

    Larsen, K S; Eydal, M; Mencke, N; Sigurdsson, H

    2005-08-01

    Lice infestations on horses caused by the lice Werneckiella (Damalinia) equi and Haematopinus equi are observed worldwide. In this study, the distribution and clinical manifestations of lice on Icelandic horses were examined. Thirty-eight out of 93 animals (40.86%) were identified as infested with W. equi. Sixty-eight animals (73.12%) presented dermatological lesions associated with lice infestation, while only 32 of these animals presented lice. Six animals had no clinical signs although of being lice-positive, and 19 animals (20.43%) showed neither lice nor clinical manifestations. Lice burdens varied from animal to animal, and clinical manifestations occurred at all levels of infestation. Focal alopecia was the main clinical sign (83.78%) on lice-positive horses, while scaling and crusts occurred in 10.81% and 9.68% of the cases, respectively. Clinical signs present in the head and the neck/mane area were found to be an indication of lice infestation in horses.

  14. Increasing awareness of Rhodococcus equi pulmonary infection in the immunocompetent adult: a rare infection with poor prognosis.

    PubMed

    Herath, Samantha; Lewis, Christopher; Nisbet, Mitzi

    2013-12-13

    The aim of this case report and review is to increase awareness of this uncommon infection with Rhodococcus equi (R. equi), in immunocompetent adults. R. equi is a soil-dwelling Gram-positive bacillus that frequently causes infection in grazing livestock. Human infection is rare and mostly limited to the immunocompromised hosts. We present a case of pneumonia caused by R. equi infection in a 55-year-old male builder who presented with cough, dyspnoea and night sweats, initially suspected to have pulmonary tuberculosis. Following biopsy of the mediastinal lymph nodes, R. equi was cultured, which is usually not a contaminant. Despite extensive investigations a host immune defect was not identified. The patient recovered after three months of combination antibiotic treatment, initially with intravenous vancomycin and meropenem followed by oral clarithromycin and rifampicin. To further clarify this rare disease we did a literature review that identified 26 adult patients with R. equi infection, without an identified host immunosuppressive condition. In this cohort, the median age at presentation was 53 years and infection holds a strong male predominance 19 (73%). An environmental exposure (e.g. farming, horse breeder) was found in 13 (50%). Ten (38%) of these patients had pulmonary infection. All deaths 3 (12%) occurred in the patients had pulmonary infection. R.equi is an infection that is difficult to diagnose and carries a high mortality if prompt treatment is not established. It is important to realise the potential for this disease to be misdiagnosed as pulmonary tuberculosis or community acquired pneumonia. Clinical suspicion is important especially if an environmental exposure is suspected.

  15. Assessment of Theileria equi and Babesia caballi infections in equine populations in Egypt by molecular, serological and hematological approaches.

    PubMed

    Mahmoud, Mona S; El-Ezz, Nadia T Abu; Abdel-Shafy, Sobhy; Nassar, Somia A; El Namaky, Amira H; Khalil, Wagdy K B; Knowles, Don; Kappmeyer, Lowell; Silva, Marta G; Suarez, Carlos E

    2016-05-04

    Equine piroplasmosis (EP) caused by Theileria equi, Babesia caballi, or both, contributes to significant economic loss in the equine industry and remains uncontrolled in Egypt. This study focuses on surveying T. equi and B. caballi infections and hematological disorders in equine populations in Egypt. Theileria equi and B. caballi infections were assessed in blood from 88 horses and 51 donkeys in Egypt using light microscopy, indirect immunofluorescent antibody test (IFAT), nested PCR (nPCR), and competitive-ELISA (cELISA) assays. PCR products were examined for specificity by DNA sequencing. Hematological alterations were evaluated using a standard cell counter. Microscopic analysis revealed EP infection in 11.4% and 17.8% of horses and donkeys respectively. IFAT detected 23.9% and 17.0% infection of T. equi and B. caballi, respectively, in horses, and 31.4% of T. equi and B. caballi in donkeys. T. equi cELISA detected 14.8% and 23.5% positive horses and donkeys, respectively, but the B. caballi RAP-1-based cELISA failed to detect any positives, a result hypothesized to be caused by sequence polymorphism found in the rap-1 genes. Nested-PCR analysis identified 36.4% and 43.1% positive horses and donkeys, respectively for T. equi and it also identified 19.3% and 15.7% positive horses and donkeys, respectively for B. caballi. The overall EP incidence found in the population under study was relatively high and comparable regardless of the diagnostic method used (56.8% using nPCR and 48.9% using IFAT). Hematologic analysis revealed macrocytic hypochromic anemia and thrombocytopenia in all piroplasma-infected horses. The data confirm relatively high levels of EP, likely causing hematological abnormalities in equines in Egypt, and also suggest the need for an improved serological test to diagnose B. caballi infection in this region.

  16. Effects of age and macrophage lineage on intracellular survival and cytokine induction after infection with Rhodococcus equi.

    PubMed

    Berghaus, Londa J; Giguère, Steeve; Sturgill, Tracy L

    2014-07-15

    Rhodococcus equi, a facultative intracellular pathogen of macrophages, causes life-threatening pneumonia in foals and in people with underlying immune deficiencies. As a basis for this study, we hypothesized that macrophage lineage and age would affect intracellular survival of R. equi and cytokine induction after infection. Monocyte-derived and bronchoalveolar macrophages from 10 adult horses and from 10 foals (sampled at 1-3 days, 2 weeks, 1 month, 3 months, and 5 months of age) were infected ex vivo with virulent R. equi. Intracellular R. equi were quantified and mRNA expression of IL-1β, IL-4, IL-6, IL-8, IL-10, IL-12 p40, IL-18, IFN-γ, and TNF-α was measured. Intracellular replication of R. equi was significantly (P<0.001) greater in bronchoalveolar than in monocyte-derived macrophages, regardless of age. Regardless of the macrophage lineage, replication of R. equi was significantly (P=0.002) higher in 3-month-old foals than in 3-day old foals, 2-week-old foals, 1-month-old foals, and adult horses. Expression of IL-4 mRNA was significantly higher in monocyte-derived macrophages whereas expression of IL-6, IL-18, and TNF-α was significantly higher in bronchoalveolar macrophages. Induction of IL-1β, IL-10, IL-12 p40, and IL-8 mRNA in bronchoalveolar macrophages of 1-3-day old foals was significantly higher than in older foals or adult horses. Preferential intracellular survival of R. equi in bronchoalveolar macrophages of juvenile horses may play a role in the pulmonary tropism of the pathogen and in the window of age susceptibility to infection.

  17. Rhodoccocus Equi Pneumonia and Paradoxical Immune Reconstitution Inflammatory Syndrome in a Patient with Acquired Immune Deficiency Syndrome (AIDS)

    PubMed Central

    Zijoo, Ritika; Dirweesh, Ahmed; Karabulut, Nigahus

    2017-01-01

    Patient: Male, 47 Final Diagnosis: Rhodococcus equi pneumonia • paradoxical immune reconstitution inflammatory syndrome Symptoms: Cough • fever • shorthness of breath Medication: — Clinical Procedure: — Specialty: Infectious Diseases Objective: Rare co-existance of disease or pathology Background: Pulmonary infections are a major cause of mortality and morbidity in patients infected with human immunodeficiency virus (HIV) and can progress rapidly to respiratory failure and death without appropriate therapy. Herein, we present a rare case of an advanced HIV infection and Rhodoccocus equi (R. equi) pneumonia in a young male who had severe paradoxical immune reconstitution inflammatory syndrome (IRIS). Case Report: A 47-year-old nonsmoking Hispanic man with advanced HIV infection presented with severe acute necrotizing pneumonia secondary to R. equi. Although his initial response to antimicrobial therapy was optimal, he became symptomatic again in spite of continuation of antibiotics as he developed severe paradoxical IRIS 3 weeks after starting a new highly active anti-retroviral therapy (HAART). Conclusions: The diagnosis of IRIS remains challenging because of the wide variations in the clinical presentation and etiologies. In spite of its rarity as an opportunistic pathogen, we recommend that R. equi, an intracellular pathogen, be included in the differential list of pathogens associated with IRIS. PMID:28100903

  18. Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks.

    PubMed

    Fernandes, M C; Takai, S; Leite, D S; Pinto, J P A N; Brandão, P E; Santarém, V A; Listoni, F J P; Da Silva, A V; Ribeiro, M G

    2013-01-01

    The identification of pathogens of viral (Rotavirus, Coronavirus), parasitic (Toxocara spp.) and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi) origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5%) strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene) or intermediately virulent (vapB gene) profiles. Sixty-three (31.5%) strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%), A (25.4%) and B2 (22.2%). No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places.

  19. A 3-D airway epithelial cell and macrophage co-culture system to study Rhodococcus equi infection.

    PubMed

    Schwab, Ute; Caldwell, Shannon; Matychak, Mary-Beth; Felippe, Julia

    2013-07-15

    We developed a 3-D equine bronchial epithelial cell (BEC) culture that fully differentiates into ciliary beating and mucus producing cells. Using this system, we evaluated how mucus affects the phagocytic activity of macrophages. Adult horse monocyte-derived macrophages were incubated with Rhodococcus equi for 4h either in the mucus layer of in vitro generated airway epithelium or on collagen coated membranes. Using light and electron microscopy, we noted that the number of macrophages with intracellular bacteria, and the number of intracellular bacteria per macrophage were lower in the presence of mucus. TNFα measurements revealed that the presence of BECs promoted TNFα production by R. equi-infected macrophages; a decrease in TLR-2 (involved in R. equi recognition) and an increase in EGF-R (involved in mucin production) mRNA expression were also noted. Interestingly, when foal macrophages were added to foal BECs, we made the opposite observation, i.e. many macrophages were loaded with R. equi. Our in vitro bronchial system shows great potential for the identification of mechanisms how BECs and mucus play a role in phagocyte activation and bacterial clearance. Further studies using this system will show whether the airway environment in the foal responds differently to R. equi infection.

  20. Evaluation of a commercially available hyperimmune plasma product for prevention of naturally acquired pneumonia caused by Rhodococcus equi in foals.

    PubMed

    Giguère, Steeve; Gaskin, Jack M; Miller, Corey; Bowman, James L

    2002-01-01

    To determine efficacy of a commercially available hyperimmune plasma product for prevention of naturally acquired pneumonia caused by Rhodococcus equi in foals. Randomized clinical trial. 165 foals. Foals were randomly assigned to 1 of 2 groups (hyperimmune plasma or nontreated controls). Foals with failure of passive transfer (FPT) of immunity were treated with hyperimmune plasma and evaluated as a third group. Foals that received plasma were given 950 ml between 1 and 10 days of age and between 30 and 50 days of age. A tracheobronchial aspirate was obtained from foals with clinical signs of respiratory tract disease for bacteriologic culture. A significant difference in incidence of pneumonia caused by R equi in foals with adequate passive transfer was not detected between foals that received plasma (19.1%) and nontreated foals (30%). Of 13 foals without FPT that received plasma and developed pneumonia caused by R equi, 12 developed disease prior to administration of the second dose of hyperimmune plasma. Incidence of undifferentiated pneumonia of all causes was not different between groups. Intravenous administration of the commercially available hyperimmune plasma was safe, and the product contained high concentrations of anti-R equi antibodies. However, within this limited foal population, the difference in incidence of pneumonia caused by R equi observed between foals that received plasma and control foals was not significant.

  1. Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks

    PubMed Central

    Fernandes, M.C.; Takai, S.; Leite, D.S.; Pinto, J.P.A.N.; Brandão, P.E.; Santarém, V.A.; Listoni, F.J.P.; Da Silva, A.V.; Ribeiro, M.G.

    2013-01-01

    The identification of pathogens of viral (Rotavirus, Coronavirus), parasitic (Toxocara spp.) and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi) origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5%) strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene) or intermediately virulent (vapB gene) profiles. Sixty-three (31.5%) strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%), A (25.4%) and B2 (22.2%). No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places. PMID:24294244

  2. Detection of Theileria equi and Babesia caballi using microscopic and molecular methods in horses in suburb of Urmia, Iran.

    PubMed

    Malekifard, Farnaz; Tavassoli, Mousa; Yakhchali, Mohammad; Darvishzadeh, Reza

    2014-01-01

    Equine piroplasmosis is a severe disease of horses caused by the intra-erythrocyte protozoan, Theileria equi and Babesia caballi. The aim of this study was to identify equine piroplasmosis based on molecular and morphometrical features in horses in suburb of Urmia, West Azerbaijan province, Iran. From April to September 2011, a total number of 240 blood samples were collected randomly from horses of 25 villages. The specimens were transferred to the laboratory and the blood smears stained with Geimsa, and the morphological and biometrical data of parasite in any infected erythrocyte were considered. Extracted DNA from each blood sample was used in multiplex PCR in order to confirm the presence of B. caballi and T. equi. Microscopic observation on 240 blood smears determined that 15 (6.25%) and 5 (2.80%) samples were infected by T. equi and B. caballi, respectively. The mixed infections occurred in 2 (0.83%) samples. The results of the PCR assays showed 26 (10.83%), 14 (5.83%) and 4 (1.66%) were distinguished as T. equi, B. caballi and mixed infection, respectively. Differences in infection rates were statistically nonsignificant between male and female horses and among different age groups. Our findings indicated that T. equi and B. caballi were prevalent in horse population.

  3. Characterization of Rhodococcus equi isolates from submaxillary lymph nodes of wild boars (Sus scrofa), red deer (Cervus elaphus) and roe deer (Capreolus capreolus).

    PubMed

    Rzewuska, Magdalena; Witkowski, Lucjan; Cisek, Agata A; Stefańska, Ilona; Chrobak, Dorota; Stefaniuk, Elżbieta; Kizerwetter-Świda, Magdalena; Takai, Shinji

    2014-08-06

    Rhodococcus equi is a soil saprophyte and an opportunistic pathogen causing infections in animals, and rarely in humans. The presence of R. equi in tissues and faeces of some wild animal species was demonstrated previously. In this study we characterized R. equi isolates from submaxillary lymph nodes of free-living wild boars (n=23), red deer (n=2) and roe deer (n=2). This is the first description of R. equi strains isolated from tissues of the Cervidae. All isolates were initially recognized as R. equi based on the phenotypic properties. Their identification was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, detection of the choE gene and by sequence analysis of the 16S rRNA and rpoB genes. The presence of three plasmidic genes (traA, vapA and vapB) associated with R. equi virulence was investigated by PCR. In 16 wild boar isolates the traA and vapB genes were detected and they were located on virulence plasmids type 5, 7 or 11. The isolates from cervids and the remaining wild boar isolates were classified as avirulent based on a genotype traA(-)/vapA(-)B(-). In summary, these results confirm that wild boars can be a source of intermediately virulent R. equi strains, and indicate that red deer and roe deer can be a reservoir of avirulent R. equi strains. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Association of airborne concentration of virulent Rhodococcus equi with location (stall versus paddock) and month (January through June) on 30 horse breeding farms in central Kentucky.

    PubMed

    Cohen, Noah D; Kuskie, Kyle R; Smith, Jacqueline L; Slovis, Nathan M; Brown, Stuart E; Stepusin, Randolph S; Chaffin, M Keith; Takai, Shinji; Carter, Craig N

    2012-10-01

    To determine whether the concentration of airborne virulent Rhodococcus equi varied by location (stall vs paddock) and month on horse farms. Air samples from stalls and paddocks used to house mares and foals on 30 horse breeding farms in central Kentucky. Air samples from 1 stall and 1 paddock were obtained monthly from each farm from January through June 2009. Concentrations of airborne virulent R equi were determined via a modified colony immunoblot assay. Random-effects logistic regression was used to determine the association of the presence of airborne virulent R equi with location from which air samples were obtained and month during which samples were collected. Of 180 air samples, virulent R equi was identified in 49 (27%) and 13 (7%) obtained from stalls and paddocks, respectively. The OR of detecting virulent R equi in air samples from stalls versus paddocks was 5.2 (95% confidence interval, 2.1 to 13.1). Of 60 air samples, virulent R equi was identified in 25 (42%), 18 (30%), and 6 (10%) obtained from stalls during January and February, March and April, and May and June, respectively. The OR of detecting virulent R equi from stall air samples collected during May and June versus January and February was 0.22 (95% confidence interval, 0.08 to 0.63). Foals were more likely to be exposed to airborne virulent R equi when housed in stalls versus paddocks and earlier (January and February) versus later (May and June) during the foaling season.

  5. Prevalence of Streptococcus equi subsp. equi in horses and associated risk factors in the State of Rio Grande do Sul, Brazil.

    PubMed

    Libardoni, Felipe; Machado, Gustavo; Gressler, Letícia Trevisan; Kowalski, Ananda Paula; Diehl, Gustavo Nogueira; dos Santos, Lucila Carboneiro; Corbellini, Luis Gustavo; de Vargas, Agueda Castagna

    2016-02-01

    The aim of this study was to estimate the prevalence of equine strangles and to identify associated risk factors for this disease through a cross-sectional study of nasal swabs. Nasal swabs (n=1010) from healthy equines (absence of nasal discharge, lymphadenopathy and cough) from 341 farms were plated on 5% blood agar; of these horses, 24 were identified as positive for Streptococcus equi through isolation, PCR and DNA sequencing. The estimated prevalence for individual animals was 2.3%, and for herds, it was 5.86%. Statistical analysis identified the following as associated risk factors: the number of group events that were attended by the equines (PR: 1.06); the sharing of food containers (PR: 3.74); and at least one previous positive diagnosis of strangles on the farm (PR: 3.20). These results constitute an epidemiological contribution to the horse industry and may support measures for the future control of the disease.

  6. Age-related changes following in vitro stimulation with Rhodococcus equi of peripheral blood leukocytes from neonatal foals.

    PubMed

    Kachroo, Priyanka; Ivanov, Ivan; Seabury, Ashley G; Liu, Mei; Chowdhary, Bhanu P; Cohen, Noah D

    2013-01-01

    Rhodococcus equi is an intracellular bacterium primarily known as an equine pathogen that infects young foals causing a pyogranulomatuous pneumonia. The molecular mechanisms mediating the immune response of foals to R. equi are not fully elucidated. Hence, global genomic high-throughput tools like gene expression microarrays might identify age-related gene expression signatures and molecular pathways that contribute to the immune mechanisms underlying the inherent susceptibility of foals to disease caused by R. equi. The objectives of this study were 2-fold: 1) to compare the expression profiles at specific ages of blood leukocytes from foals stimulated with virulent R. equi with those of unstimulated leukocytes; and, 2) to characterize the age-related changes in the gene expression profile associated with blood leukocytes in response to stimulation with virulent R. equi. Peripheral blood leukocytes were obtained from 6 foals within 24 hours (h) of birth (day 1) and 2, 4, and 8 weeks after birth. The samples were split, such that half were stimulated with live virulent R. equi, and the other half served as unstimulated control. RNA was extracted and the generated cDNA was labeled with fluorescent dyes for microarray hybridizations using an equine microarray. Our findings suggest that there is age-related differential expression of genes involved in host immune response and immunity. We found induction of genes critical for host immunity against pathogens (MHC class II) only at the later time-points (compared to birth). While it appears that foals up to 8-weeks of age are able to initiate a protective inflammatory response against the bacteria, relatively decreased expression of various other immune-related genes points toward inherent diminished immune responses closer to birth. These genes and pathways may contribute to disease susceptibility in foals if infected early in life, and might thus be targeted for developing preventative or therapeutic strategies.

  7. Policies and Processes for Social Inclusion: Using EquiFrame and EquIPP for Policy Dialogue

    PubMed Central

    MacLachlan, Malcolm; Mannan, Hasheem; Huss, Tessy; Munthali, Alister; Amin, Mutamad

    2016-01-01

    The application of EquiFrame in the analysis of sexual and reproductive health policies by Ivanova et al to a new thematic area, their selection of only some of the Core Concepts of human rights in health service provision and the addition of new vulnerable groups relevant to the purpose of their analysis, are all very welcome developments. We also applaud their application of EquiFrame to policies in countries where it has not previously been used, along with their use of interviews with policy-makers to produce a deeper understanding of policy processes. We argue that clear justification for the inclusion of additional, or replacement of some exiting vulnerable groups within EquiFrame should be accompanied by clear definitions of such groups, along with the evidence-base that justifies their classification as a vulnerable or marginalised group. To illustrate the versatility of EquiFrame, we summarise a range of ways in which it has been used across a number of regions; including a brief Case Study of its use to develop the National Health Policy of Malawi. While EquiFrame focuses on policy content, we preview a new policy analysis tool – Equity and Inclusion in Policy Processes (EquIPP) – which assesses the extent of equity and inclusion in broader policy processes. Together, EquiFrame and EquIPP can be used to help governments and civil society ensure that policies are addressing the much stronger emphasis on social inclusion, now apparent in the Sustainable Development Goals (SDGs). PMID:26927591

  8. Affects of N-terminal variation in the SeM protein of Streptococcus equi on antibody and fibrinogen binding.

    PubMed

    Timoney, John F; DeNegri, Rafaela; Sheoran, Abhineet; Forster, Nathalie

    2010-02-10

    The clonal Streptococcus equi causes equine strangles, a highly contagious suppurative lymphadenopathy and rhinopharyngitis. An important virulence factor and vaccine component, the antiphagocytic fibrinogen binding SeM of S. equi is a surface anchored fibrillar protein. Two recent studies of N. American, Japanese and European isolates have revealed a high frequency of N-terminal amino acid variation in SeM of S. equi CF32 that suggests this region of the protein is subject to immunologic selection pressure. The aims of the present study were firstly to map regions of SeM reactive with convalescent equine IgG and IgA and stimulatory for lymph node cells and secondly to determine effects of N-terminal variation on the functionality of SeM. Variation did not significantly affect fibrinogen binding or susceptibility of S. equi to an opsonic equine serum. Linear epitopes reactive with convalescent IgG and mucosal IgA were concentrated toward the conserved center of SeM. However, IgA but not IgG from every horse reacted with at least one peptide that contained variable sequence. Lymph node cells (CD4+) from horses immunized with SeM were strongly responsive to a peptide (alphaalpha36-138) encoding the entire variable region. SeM (CF32) specific mouse Mab 04D11 which reacted strongly with this larger peptide but not with shorter peptides within that sequence reacted strongly with whole cells of S. equi CF32 but only weakly with cells of any of 14 isolates of S. equi expressing different variants of SeM. These results in combination suggest that N-terminal variation alters a conformational epitope of significance in mucosal IgA and systemic T cell responses but does not affect antibody mediated phagocytosis and killing. Copyright (c) 2009 Elsevier Ltd. All rights reserved.

  9. The Steroid Catabolic Pathway of the Intracellular Pathogen Rhodococcus equi Is Important for Pathogenesis and a Target for Vaccine Development

    PubMed Central

    van der Geize, R.; Grommen, A. W. F.; Hessels, G. I.; Jacobs, A. A. C.; Dijkhuizen, L.

    2011-01-01

    Rhodococcus equi causes fatal pyogranulomatous pneumonia in foals and immunocompromised animals and humans. Despite its importance, there is currently no effective vaccine against the disease. The actinobacteria R. equi and the human pathogen Mycobacterium tuberculosis are related, and both cause pulmonary diseases. Recently, we have shown that essential steps in the cholesterol catabolic pathway are involved in the pathogenicity of M. tuberculosis. Bioinformatic analysis revealed the presence of a similar cholesterol catabolic gene cluster in R. equi. Orthologs of predicted M. tuberculosis virulence genes located within this cluster, i.e. ipdA (rv3551), ipdB (rv3552), fadA6 and fadE30, were identified in R. equi RE1 and inactivated. The ipdA and ipdB genes of R. equi RE1 appear to constitute the α-subunit and β-subunit, respectively, of a heterodimeric coenzyme A transferase. Mutant strains RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, were impaired in growth on the steroid catabolic pathway intermediates 4-androstene-3,17-dione (AD) and 3aα-H-4α(3′-propionic acid)-5α-hydroxy-7aβ-methylhexahydro-1-indanone (5α-hydroxy-methylhexahydro-1-indanone propionate; 5OH-HIP). Interestingly, RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, also displayed an attenuated phenotype in a macrophage infection assay. Gene products important for growth on 5OH-HIP, as part of the steroid catabolic pathway, thus appear to act as factors involved in the pathogenicity of R. equi. Challenge experiments showed that RE1ΔipdAB could be safely administered intratracheally to 2 to 5 week-old foals and oral immunization of foals even elicited a substantial protective immunity against a virulent R. equi strain. Our data show that genes involved in steroid catabolism are promising targets for the development of a live-attenuated vaccine against R. equi infections. PMID:21901092

  10. The steroid catabolic pathway of the intracellular pathogen Rhodococcus equi is important for pathogenesis and a target for vaccine development.

    PubMed

    van der Geize, R; Grommen, A W F; Hessels, G I; Jacobs, A A C; Dijkhuizen, L

    2011-08-01

    Rhodococcus equi causes fatal pyogranulomatous pneumonia in foals and immunocompromised animals and humans. Despite its importance, there is currently no effective vaccine against the disease. The actinobacteria R. equi and the human pathogen Mycobacterium tuberculosis are related, and both cause pulmonary diseases. Recently, we have shown that essential steps in the cholesterol catabolic pathway are involved in the pathogenicity of M. tuberculosis. Bioinformatic analysis revealed the presence of a similar cholesterol catabolic gene cluster in R. equi. Orthologs of predicted M. tuberculosis virulence genes located within this cluster, i.e. ipdA (rv3551), ipdB (rv3552), fadA6 and fadE30, were identified in R. equi RE1 and inactivated. The ipdA and ipdB genes of R. equi RE1 appear to constitute the α-subunit and β-subunit, respectively, of a heterodimeric coenzyme A transferase. Mutant strains RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, were impaired in growth on the steroid catabolic pathway intermediates 4-androstene-3,17-dione (AD) and 3aα-H-4α(3'-propionic acid)-5α-hydroxy-7aβ-methylhexahydro-1-indanone (5α-hydroxy-methylhexahydro-1-indanone propionate; 5OH-HIP). Interestingly, RE1ΔipdAB and RE1ΔfadE30, but not RE1ΔfadA6, also displayed an attenuated phenotype in a macrophage infection assay. Gene products important for growth on 5OH-HIP, as part of the steroid catabolic pathway, thus appear to act as factors involved in the pathogenicity of R. equi. Challenge experiments showed that RE1ΔipdAB could be safely administered intratracheally to 2 to 5 week-old foals and oral immunization of foals even elicited a substantial protective immunity against a virulent R. equi strain. Our data show that genes involved in steroid catabolism are promising targets for the development of a live-attenuated vaccine against R. equi infections.

  11. Efficacy of buparvaquone as a therapeutic and clearing agent of Babesia equi of European origin in horses.

    PubMed

    Zaugg, J L; Lane, V M

    1992-08-01

    We evaluated the efficacy of buparvaquone in eliminating infection with Babesia equi of European origin in carrier horses and in splenectomized horses with experimentally induced acute infection. When administered at the rate of 5 mg/kg of body weight, IV, 4 times at 48-hour intervals, buparvaquone prompted rapid abatement of parasitemia. However, secondary and tertiary recrudescent parasitemias invariably returned with establishment of the carrier state. Buparvaquone, at the dosage evaluated, had transitory therapeutic efficacy against acute B equi infection in splenectomized horses, but was unable alone to clear carrier infection.

  12. Gallium maltolate as an alternative to macrolides for treatment of presumed Rhodococcus equi pneumonia in foals.

    PubMed

    Cohen, Noah D; Slovis, Nathan M; Giguère, Steeve; Baker, Samantha; Chaffin, M Keith; Bernstein, Lawrence R

    2015-01-01

    Macrolide-resistant isolates of Rhodococcus equi are emerging, prompting the search for clinically effective alternative antimicrobials. The proportion of foals with ultrasonographic evidence of pneumonia presumed to be caused by R. equi that had a successful outcome when administered gallium maltolate (GaM) PO would not be more than 10% inferior (ie, lower) than that of foals receiving standard treatment. Fifty-four foals with subclinical pulmonary abscesses among 509 foals at 6 breeding farms in Kentucky. Controlled, randomized, prospective noninferiority study. Foals with ultrasonographic lesions >1 cm in diameter (n = 54) were randomly allocated to receive per os either clarithromycin combined with rifampin (CLR+R) or GaM, and followed up for 28 days by daily physical inspections and weekly (n = 1 farm) or biweekly (n = 4 farms) thoracic ultrasound examinations by individuals unaware of treatment-group assignments. Treatment success was defined as resolution of ultrasonographically identified pulmonary abscesses within 28 days of initiating treatment. Noninferiority was defined as a 90% confidence interval for the observed difference in CLR+R minus GaM that was ≤10%. The proportion of GaM-treated foals that resolved (70%; 14/20) was similar to that of foals treated with CLR+R (74%; 25/34), but we failed to demonstrate noninferiority for GaM relative to CLR+R; however, GaM was noninferior to CLR+R treatment when results from a noncompliant farm were excluded. Gallium maltolate is not inferior to macrolides for treating foals with subclinical pneumonia. Use of GaM might reduce pressure for macrolide-resistance in R. equi. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  13. Complex unconstrained three-dimensional hand movement and constant equi-affine speed.

    PubMed

    Maoz, Uri; Berthoz, Alain; Flash, Tamar

    2009-02-01

    One long-established simplifying principle behind the large repertoire and high versatility of human hand movements is the two-thirds power law-an empirical law stating a relationship between local geometry and kinematics of human hand trajectories during planar curved movements. It was further generalized not only to various types of human movements, but also to motion perception and prediction, although it was unsuccessful in explaining unconstrained three-dimensional (3D) movements. Recently, movement obeying the power law was proved to be equivalent to moving with constant planar equi-affine speed. Generalizing such motion to 3D space-i.e., to movement at constant spatial equi-affine speed-predicts the emergence of a new power law, whose utility for describing spatial scribbling movements we have previously demonstrated. In this empirical investigation of the new power law, subjects repetitively traced six different 3D geometrical shapes with their hand. We show that the 3D power law explains the data consistently better than both the two-thirds power law and an additional power law that was previously suggested for spatial hand movements. We also found small yet systematic modifications of the power-law's exponents across the various shapes, which further scrutiny suggested to be correlated with global geometric factors of the traced shape. Nevertheless, averaging over all subjects and shapes, the power-law exponents are generally in accordance with constant spatial equi-affine speed. Taken together, our findings provide evidence for the potential role of non-Euclidean geometry in motion planning and control. Moreover, these results seem to imply a relationship between geometry and kinematics that is more complex than the simple local one stipulated by the two-thirds power law and similar models.

  14. Estimating the Sensitivity and Specificity of Real-Time Quantitative PCR of Fecal Samples for Diagnosis of Rhodococcus equi Pneumonia in Foals.

    PubMed

    Shaw, S D; Cohen, N D; Chaffin, M K; Blodgett, G P; Syndergaard, M; Hurych, D

    2015-01-01

    Real-time, quantitative PCR (qPCR) methods for detecting Rhodococcus equi in feces have been developed as a noninvasive, rapid diagnostic test for R. equi pneumonia, but have not been evaluated in a large population of foals. The objective of this study was to evaluate the clinical utility of fecal PCR as a diagnostic test for R. equi pneumonia in foals using receiver operating characteristic (ROC) methods. 186 foals born in 2011 at an R. equi-endemic ranch in Texas. Fecal samples were collected at the time of onset of clinical signs for pneumonic foals (n = 31). Foals with pneumonia were matched by age and birth date to healthy (n = 31) and subclinical (n = 124) control foals; fecal samples were collected from these controls. DNA was extracted from feces using commercial kits and concentration of virulent R. equi in feces was determined by qPCR. Concentration of R. equi in feces differed significantly (P < .05) among groups. The area under the ROC curve for fecal qPCR for diagnosis of R. equi pneumonia was 89% (95% CI, 83-99), with a sensitivity of 94% and specificity of 72%. qPCR of feces can be useful as an alternative to tracheobronchial aspiration for the diagnosis of R. equi in foals with clinical signs of pneumonia. Caution should be used in extrapolating results of this study to other populations because fecal concentration of R. equi might vary by geographic location or management practices. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  15. Inhibitory effect of allicin on the growth of Babesia and Theileria equi parasites.

    PubMed

    Salama, Akram Ahmed; AbouLaila, Mahmoud; Terkawi, Mohamad Alaa; Mousa, Ahmed; El-Sify, Ahmed; Allaam, Mahmoud; Zaghawa, Ahmed; Yokoyama, Naoaki; Igarashi, Ikuo

    2014-01-01

    Allicin is an active ingredient of garlic that has antibacterial, antifungal, antiviral, and antiprotozoal activity. However, the inhibitory effects of allicin on Babesia parasites have not yet been examined. In the present study, allicin was tested as a potent inhibitor against the in vitro growth of bovine and equine Babesia parasites and the in vivo growth of Babesia microti in a mouse model. The in vitro growth of Babesia bovis, Babesia bigemina, Babesia caballi, or Theileria equi was inhibited by allicin in a dose-dependent manner and had IC50 values of 818, 675, 470, and 742 μM, respectively. Moreover, allicin significantly inhibited (P < 0.001) invasion of B. bovis, B. bigemina, B. caballi, and T. equi into the host erythrocyte. Furthermore, mice treated with 30 mg/kg of allicin for 5 days significantly (P < 0.05) reduced the parasitemia of B. microti over the period of the study. To further examine the potential synergism of allicin with diminazene aceturate, growth inhibitory assays were performed in vitro and in vivo. Interestingly, combinations of diminazene aceturate with allicin synergistically potentiated its inhibitory effects in vitro and in vivo. These results indicate that allicin might be beneficial for the treatment of babesiosis, particularly when used in combination with diminazene aceturate.

  16. Lymphocyte stimulation response in horses against phytohaemagglutinin and M protein of Streptococcus equi using whole blood.

    PubMed Central

    Srivastava, S K; Barnum, D A

    1982-01-01

    Lymphocyte stimulation was observed in whole equine blood in the presence of phytohaemagglutinin and M protein extracted from a typical strain of Streptococcus equi. Blood samples were collected from several healthy horses and horse and pony foals and cultured in vitro with varying concentrations of phytohaemagglutinin and M protein for several days. Phytohaemagglutinin was found to induce lymphocyte stimulation in these animals. Highest mean stimulation indices in horse foals (49.3 +/- 24.4) and pony foals (54.7 +/- 32.0) were observed with 0.625 and 1.25 micrograms/mL phytohaemagglutinin, respectively, at either 72 or 96 hours of incubation. Significantly higher radioactive counts per minute in horse and pony foals were recorded in blood cultures incubated with 0.625 and 1.25 micrograms/mL phytohaemagglutinin. M protein induced a dose related stimulation response in adult horses. Maximum stimulation indices were observed against 125 micrograms/mL M protein at 96 hours. These stimulation indices were higher in adult horses (40.0 +/- 2.2) than observed in pony foals (14.4 +/- 15.7). Higher stimulation levels in adult horses indicated either nonspecific stimulation against M protein or previous exposure of these animals to S. equi. PMID:7074416

  17. Passive transfer of mucosal antibody to Streptococcus equi in the foal.

    PubMed Central

    Galan, J E; Timoney, J F; Lengemann, F W

    1986-01-01

    Passive transfer of mucosal antibody to Streptococcus equi was studied in foals during the first 2 months of life. Immunoglobulin G (IgG) and IgA antibodies were found in sera and nasal secretions of foals shortly after colostrum intake. Titers were highest 2 days after birth; IgG predominated in sera, and IgA predominated in nasal washes. Intragastrically administered 99mTc-labeled IgA was transported from the bloodstream to the nasal mucosa of a newborn foal within a few hours of colostrum intake. Western blot analysis of the specificities of colostral and serum antibodies showed that selective transfer of immunoglobulins of defined specificity did not occur. Antibodies from milk samples taken a month or more into lactation had different specificities than those of colostrum or serum samples. Acid-extracted M protein fragments of S. equi recognized by milk antibodies were the same as those recognized by IgG and IgA from nasopharyngeal mucus of horses recently recovered from strangles. We postulate that passive antibody protection of the foal is derived both by secretion of colostral immunoglobulins onto the nasopharyngeal mucosa and by immunoglobulins ingested in milk that directly coat the upper respiratory and oral mucosa during the first months of life. Images PMID:3531013

  18. Passive transfer of mucosal antibody to Streptococcus equi in the foal.

    PubMed

    Galan, J E; Timoney, J F; Lengemann, F W

    1986-10-01

    Passive transfer of mucosal antibody to Streptococcus equi was studied in foals during the first 2 months of life. Immunoglobulin G (IgG) and IgA antibodies were found in sera and nasal secretions of foals shortly after colostrum intake. Titers were highest 2 days after birth; IgG predominated in sera, and IgA predominated in nasal washes. Intragastrically administered 99mTc-labeled IgA was transported from the bloodstream to the nasal mucosa of a newborn foal within a few hours of colostrum intake. Western blot analysis of the specificities of colostral and serum antibodies showed that selective transfer of immunoglobulins of defined specificity did not occur. Antibodies from milk samples taken a month or more into lactation had different specificities than those of colostrum or serum samples. Acid-extracted M protein fragments of S. equi recognized by milk antibodies were the same as those recognized by IgG and IgA from nasopharyngeal mucus of horses recently recovered from strangles. We postulate that passive antibody protection of the foal is derived both by secretion of colostral immunoglobulins onto the nasopharyngeal mucosa and by immunoglobulins ingested in milk that directly coat the upper respiratory and oral mucosa during the first months of life.

  19. Detection of Babesia caballi and Theileria equi in Blood from Equines from Four Indigenous Communities in Costa Rica

    PubMed Central

    Posada-Guzmán, María Fernanda; Romero-Zúñiga, Juan José; Jiménez-Rocha, Ana Eugenia

    2015-01-01

    A cross-sectional study was carried out in four indigenous communities of Costa Rica to detect presence and prevalence of Babesia caballi and Theileria equi and to investigate factors associated with presence of these hemoparasites. General condition of horses (n = 285) was evaluated, and hematocrits and hemoglobin were determined from blood samples of 130 horses, which were also analyzed using blood smears, nested polymerase chain reaction (PCR), and immunosorbent assay (c-ELISA). The general condition of the horses (n = 285) in terms of their body and coat was between regular and poor, and hematocrit and hemoglobin average values were low (19% and 10.65 g/dL, resp.). Erythrocyte inclusions were observed in 32 (24.6%) of the samples. Twenty-six samples (20.0%) gave positive results for B. caballi and 60 (46.2%) for T. equi; 10 horses (7.7%) showed mixed infection, when analyzed by PCR. Using c-ELISA, it was found that 90 (69.2%) horses had antibodies against B. caballi and 115 (88.5%) against T. equi, while 81 (62.3%) showed mixed reactions. There were no factors associated with the presence of B. caballi and T. equi. These results contrast with results previously obtained in equines in the Central Valley of Costa Rica. PMID:26649225

  20. Effects of dihydroorotate dehydrogenase (DHODH) inhibitors on the growth of Theileria equi and Babesia caballi in vitro.

    PubMed

    Kamyingkird, Ketsarin; Cao, Shinuo; Tuvshintulga, Bumduuren; Salama, Akram; Mousa, Ahmed Abdelmoniem; Efstratiou, Artemis; Nishikawa, Yoshifumi; Yokoyama, Naoaki; Igarashi, Ikuo; Xuan, Xuenan

    2017-05-01

    Theileria equi and Babesia caballi are the causative agents of equine piroplasmosis (EP), which affects equine production in various parts of the world. However, a safe and effective drug is not currently available for treatment of EP. Dihydroorotate dehydrogenase (DHODH) is the fourth enzyme in the de novo pyrimidine synthesis pathway and has been known as a novel drug target for several apicomplexan protozoan parasites. In this study, we evaluated four DHODH inhibitors; atovaquone (ATV), leflunomide (LFN), brequinar (Breq), and 7-hydroxy-5-[1,2,4] triazolo [1,5,a] pyrimidine (TAZ) on the growth of T. equi and B. caballi in vitro and compared them to diminacene aceturate (Di) as the control drug. The growth of T. equi and B. caballi was significantly hindered by all inhibitors except TAZ. The half maximal inhibitory concentration (IC50) of ATV, LFN, Breq and Di against T. equi was approximately 0.028, 109, 11 and 40 μM, respectively, whereas the IC50 of ATV, LFN, Breq and Di against B. caballi was approximately 0.128, 193, 5.2 and 16.2 μM, respectively. Using bioinformatics and Western blot analysis, we showed that TeDHODH was similar to other Babesia parasite DHODHs, and confirmed that targeting DHODHs could be useful for the development of novel chemotherapeutics for treatment of EP.

  1. Inhibition of the in vitro growth of babesia bigemina, babesia caballi and theileria equi parasites by trifluralin analogues

    USDA-ARS?s Scientific Manuscript database

    Background: Bovine and equine babesiosis caused by Babesia bovis, B. bigemina and B. caballi, and equine theileriosis caused by Theileria equi are global tick borne hemoprotozoan diseases characterized by fever, anemia, weight losses and abortions. A common feature of these diseases are transition f...

  2. Diagnosis of theileria equi infections in horses in the Azores using cELISA and nested PCR

    USDA-ARS?s Scientific Manuscript database

    Equine piroplasmosis is a tick-borne disease of equids that is often caused by the parasite Theileria equi. We applied competitive ELISA (cELISA) and nested PCR diagnostic methods to detect this parasite in horses by screening 162 samples from mainland Portugal where the parasite is endemic, and 143...

  3. Genetic characterization of theileria equi infecting horses in North America: evidence for a limited source of U.S. introductions

    USDA-ARS?s Scientific Manuscript database

    Theileria equi is a tick-borne Apicomplexan hemoparasite that causes equine piroplasmosis (EP). This parasite has a worldwide distribution, but until recent outbreaks the United States has been considered to be free of EP. Maximum parsimony analysis of 18S rRNA gene sequences of North American T. eq...

  4. Theileria equi isolates vary in susceptibility to imidocarb dipropionate but demonstrate uniform in vitro susceptibility to a bumped kinase inhibitor

    USDA-ARS?s Scientific Manuscript database

    The apicomplexan hemoparasite Theileria equi is a causative agent of equine piroplasmosis, eradicated from the United States in 1988. However, recent outbreaks have sparked renewed interest in treatment options for infected horses. Imidocarb dipropionate is the current drug of choice, however variat...

  5. 75 FR 36456 - Channel America Television Network, Inc., EquiMed, Inc., Kore Holdings, Inc., Robotic Vision...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-25

    ... From the Federal Register Online via the Government Publishing Office SECURITIES AND EXCHANGE COMMISSION Channel America Television Network, Inc., EquiMed, Inc., Kore Holdings, Inc., Robotic Vision... concerning the securities of Robotic Vision Systems, Inc. (n/k/a Acuity Cimatrix, Inc.) because it has not...

  6. Transfer of the virulence-associated protein A-bearing plasmid between field strains of virulent and avirulent Rhodococcus equi

    USDA-ARS?s Scientific Manuscript database

    Virulent and avirulent isolates coexist in equine feces and the environment and serve as a source of infection for foals. The extent to which conjugative plasmid transfer occurs between these strains is unknown and is important for understanding the epidemiology of Rhodococcus equi infections of fo...

  7. Duddingtonia flagrans, Monacrosporium thaumasium and Pochonia chlamydosporia as possible biological control agents of Oxyuris equi and Austroxyuris finlaysoni.

    PubMed

    Braga, F R; Araújo, J V; Silva, A R; Araujo, J M; Carvalho, R O; Campos, A K; Tavela, A O; Ferreira, S R; Frassy, L N; Alves, C D F

    2010-03-01

    The action of four fungal isolates of the species Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34a) and Pochonia chlamydosporia (VC1 and VC4) on eggs of Oxyuris equi and Austroxyuris finlaysoni was evaluated in two assays (A and B). Eggs of O. equi (Test A) and A. finlaysoni (Test B) were plated on Petri dishes with 2% water-agar with grown fungal isolates and control without fungus. After 5, 10 and 15 days, 100 eggs were collected and classified according to the following parameters: type 1 effect, physiological and biochemical effect without morphological damage to the eggshell; type 2 effect, lytic effect with morphological alteration of the eggshell and embryo; and type 3 effect, lytic effect with morphological alteration of the eggshell and embryo, hyphal penetration and internal egg colonization. Pochonia chlamydosporia isolates VC1 and VC4 showed ovicidal activity for type 1, 2 and 3 effects on eggs of O. equi and eggs of A. finlaysoni. In vitro assays A and B showed that P. chlamydosporia had a negative influence on eggs of O. equi and A. finlaysoni and can be considered as a potential biological control agent of nematodes.

  8. Molecular characterization of a lipid-modified virulence-associated protein of Rhodococcus equi and its potential in protective immunity.

    PubMed Central

    Tan, C; Prescott, J F; Patterson, M C; Nicholson, V M

    1995-01-01

    Virulent strains of Rhodococcus equi produce plasmid-mediated 15- and 17-kDa proteins, which are thermoregulated and apparently surface-expressed. We demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) that R. equi produce three antigenically-related virulence-associated proteins, a diffuse 18-22-kDa, a 17.5-kDa and a 15-kDa protein. Phase partitioning of whole cells of R. equi strain 103 with Triton X-114 (TX-114) and labelling with [3H]-labelled palmitic acid showed that the two higher molecular weight proteins are hydrophobic and lipid modified. The 15-kDa protein did not partition into TX-114 and was not lipid modified. Cloning and expression of a fragment of the R. equi virulence plasmid in Escherichia coli showed that the three proteins were expressed from a single gene. Sequence analysis of this gene (designated vapA) revealed a 570-bp open reading frame encoding a polypeptide of 189 amino acids with a calculated molecular mass of 19,175 Da. The mature, nonlipid modified protein had a calculated mass of 16,246 Da. The 17.5- and 18-22-kDa forms of the protein are therefore due to lipid modification. No significant sequence homology of the vapA gene with other reported nucleotide sequences were found. Opsonization of virulent R. equi with an IgG1 mouse monoclonal antibody (MAb103) to the VapA protein significantly enhanced uptake in the murine macrophage cell line IC-21. Intraperitoneal injection of mice with Mab103 enhanced initial clearance from the liver of mice challenged intravenously with R. equi. Immunization of mice with the lipid-modified VapA purified by SDS-PAGE fractionation or with acetone precipitated VapA protein following TX-114 extraction resulted in significantly enhanced clearance from the liver and spleen following intravenous challenge. The VapA protein of R. equi appears therefore to be a protective immunogen. Images Fig. 1. Fig. 4. PMID:7704843

  9. Detection of Theileria equi and Babesia caballi infections in Venezuelan horses using Competitive-Inhibition ELISA and PCR.

    PubMed

    Rosales, Romel; Rangel-Rivas, Ariadna; Escalona, América; Jordan, Luis Segundo; Gonzatti, Mary Isabel; Aso, Pedro Maria; Perrone, Trina; Silva-Iturriza, Adriana; Mijares, Alfredo

    2013-09-01

    The focus of this study was the detection of equine piroplasmosis in Distrito Capital, Miranda, Aragua, Guárico and Apure States from Venezuela, using two methods: Competitive-Inhibition ELISA and multiplex PCR and the analysis of the possible differences in occurrence in relation to the primary purpose of the horses, which is related to varied degrees of exposure to tick. Antibody levels to Babesia caballi and Theileria equi were assessed in 694 equine serum samples using Competitive-Inhibition ELISA, while PCR assays were performed in 136 horses, using two sets of oligonucleotides to establish the presence of T. equi, B. caballi or both. The overall seroprevalence of equine piroplasmosis was 50.2%, antibodies to B. caballi were found in 161 horses (23.2%), whereas 97 (14.0%) were seropositive to T. equi and 90 (13.0%) were positives to both parasites (mixed infections). PCR determinations (n=136) showed a prevalence of 66.2%, distributed in 84 (61.8% positives) for T. equi and, 6 (4.4%) were positive to both parasites. The cELISA showed higher levels of prevalence of B. caballi and mixed infections, as compared to the PCR method. This discrepancy can be explained by the different parameters that are evaluated by each technique, PCR detect the parasite itself, while cELISA detects antibodies to the parasite. By PCR, the highest prevalence was found in Apure state, where 92.3% of the samples were positive to T. equi infections. In this locality, free grazing animals are used for livestock management. This high prevalence may be linked to the tick species present in that area. More epidemiological studies will be necessary to assess the epidemiological status of equine piroplasmosis in Venezuela. Published by Elsevier B.V.

  10. Seroprevalence and risk factors associated with Babesia caballi and Theileria equi infections in donkeys from Southern Italy.

    PubMed

    Piantedosi, D; D'Alessio, N; Di Loria, A; Di Prisco, F; Mariani, U; Neola, B; Santoro, M; Montagnaro, S; Capelli, G; Veneziano, V

    2014-12-01

    Equine piroplasmosis (EP) has been frequently described in donkeys in subtropical and tropical regions, but published data reflecting large scale surveys are very limited in Europe. The seroprevalence of Babesia caballi and Theileria equi was determined in a donkey population from Campania Region in Southern Italy using a commercial indirect fluorescent antibody test (IFAT), and the risk factors associated with the occurrence of the infection were assessed. Of 203 samples, the overall seroprevalence for EP was 57.1% (116/203), with 35.5% (72/203) for B. caballi and 44.3% (90/203) for T. equi. Co-infection was detected in 46 donkeys (22.6%). The distribution of IFAT antibody titres to B. caballi was: 1:80 (n= 67), 1:160 (n= 2), 1:320 (n= 3); while the distribution of IFAT antibody titres to T. equi was: 1:80 (n= 25), 1:160 (n= 42), 1:320 (n= 12), 1:640 (n= 8), 1:1280 (n= 3). All examined donkeys were asymptomatic, except one adult male (with a titre of 1:640 against T. equi) that showed clinical signs corresponding to the acute stage of EP, reported for the first time in Italy. The unique risk factor associated with a higher B. caballi seroprevalence was the presence of horses in the farms, while risk factors associated with a higher T. equi seroprevalence were poor body condition, presence of ruminants in the farms and milk production. The results indicate a high level of exposure in donkeys living in Southern Italy and suggest that donkeys may be an important reservoir of EP.

  11. An Invertron-Like Linear Plasmid Mediates Intracellular Survival and Virulence in Bovine Isolates of Rhodococcus equi

    PubMed Central

    Valero-Rello, Ana; Hapeshi, Alexia; Anastasi, Elisa; Alvarez, Sonsiray; Scortti, Mariela; Meijer, Wim G.; MacArthur, Iain

    2015-01-01

    We report a novel host-associated virulence plasmid in Rhodococcus equi, pVAPN, carried by bovine isolates of this facultative intracellular pathogenic actinomycete. Surprisingly, pVAPN is a 120-kb invertron-like linear replicon unrelated to the circular virulence plasmids associated with equine (pVAPA) and porcine (pVAPB variant) R. equi isolates. pVAPN is similar to the linear plasmid pNSL1 from Rhodococcus sp. NS1 and harbors six new vap multigene family members (vapN to vapS) in a vap pathogenicity locus presumably acquired via en bloc mobilization from a direct predecessor of equine pVAPA. Loss of pVAPN rendered R. equi avirulent in macrophages and mice. Mating experiments using an in vivo transconjugant selection strategy demonstrated that pVAPN transfer is sufficient to confer virulence to a plasmid-cured R. equi recipient. Phylogenetic analyses assigned the vap multigene family complement from pVAPN, pVAPA, and pVAPB to seven monophyletic clades, each containing plasmid type-specific allelic variants of a precursor vap gene carried by the nearest vap island ancestor. Deletion of vapN, the predicted “bovine-type” allelic counterpart of vapA, essential for virulence in pVAPA, abrogated pVAPN-mediated intramacrophage proliferation and virulence in mice. Our findings support a model in which R. equi virulence is conferred by host-adapted plasmids. Their central role is mediating intracellular proliferation in macrophages, promoted by a key vap determinant present in the common ancestor of the plasmid-specific vap islands, with host tropism as a secondary trait selected during coevolution with specific animal species. PMID:25895973

  12. An Invertron-Like Linear Plasmid Mediates Intracellular Survival and Virulence in Bovine Isolates of Rhodococcus equi.

    PubMed

    Valero-Rello, Ana; Hapeshi, Alexia; Anastasi, Elisa; Alvarez, Sonsiray; Scortti, Mariela; Meijer, Wim G; MacArthur, Iain; Vázquez-Boland, José A

    2015-07-01

    We report a novel host-associated virulence plasmid in Rhodococcus equi, pVAPN, carried by bovine isolates of this facultative intracellular pathogenic actinomycete. Surprisingly, pVAPN is a 120-kb invertron-like linear replicon unrelated to the circular virulence plasmids associated with equine (pVAPA) and porcine (pVAPB variant) R. equi isolates. pVAPN is similar to the linear plasmid pNSL1 from Rhodococcus sp. NS1 and harbors six new vap multigene family members (vapN to vapS) in a vap pathogenicity locus presumably acquired via en bloc mobilization from a direct predecessor of equine pVAPA. Loss of pVAPN rendered R. equi avirulent in macrophages and mice. Mating experiments using an in vivo transconjugant selection strategy demonstrated that pVAPN transfer is sufficient to confer virulence to a plasmid-cured R. equi recipient. Phylogenetic analyses assigned the vap multigene family complement from pVAPN, pVAPA, and pVAPB to seven monophyletic clades, each containing plasmid type-specific allelic variants of a precursor vap gene carried by the nearest vap island ancestor. Deletion of vapN, the predicted "bovine-type" allelic counterpart of vapA, essential for virulence in pVAPA, abrogated pVAPN-mediated intramacrophage proliferation and virulence in mice. Our findings support a model in which R. equi virulence is conferred by host-adapted plasmids. Their central role is mediating intracellular proliferation in macrophages, promoted by a key vap determinant present in the common ancestor of the plasmid-specific vap islands, with host tropism as a secondary trait selected during coevolution with specific animal species. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  13. Comparative Genomics of Rhodococcus equi Virulence Plasmids Indicates Host-Driven Evolution of the vap Pathogenicity Island.

    PubMed

    MacArthur, Iain; Anastasi, Elisa; Alvarez, Sonsiray; Scortti, Mariela; Vázquez-Boland, José A

    2017-05-01

    The conjugative virulence plasmid is a key component of the Rhodococcus equi accessory genome essential for pathogenesis. Three host-associated virulence plasmid types have been identified the equine pVAPA and porcine pVAPB circular variants, and the linear pVAPN found in bovine (ruminant) isolates. We recently characterized the R. equi pangenome (Anastasi E, et al. 2016. Pangenome and phylogenomic analysis of the pathogenic actinobacterium Rhodococcus equi. Genome Biol Evol. 8:3140-3148.) and we report here the comparative analysis of the virulence plasmid genomes. Plasmids within each host-associated type were highly similar despite their diverse origins. Variation was accounted for by scattered single nucleotide polymorphisms and short nucleotide indels, while larger indels-mostly in the plasticity region near the vap pathogencity island (PAI)-defined plasmid genomic subtypes. Only one of the plasmids analyzed, of pVAPN type, was exceptionally divergent due to accumulation of indels in the housekeeping backbone. Each host-associated plasmid type carried a unique PAI differing in vap gene complement, suggesting animal host-specific evolution of the vap multigene family. Complete conservation of the vap PAI was observed within each host-associated plasmid type. Both diversity of host-associated plasmid types and clonality of specific chromosomal-plasmid genomic type combinations were observed within the same R. equi phylogenomic subclade. Our data indicate that the overall strong conservation of the R. equi host-associated virulence plasmids is the combined result of host-driven selection, lateral transfer between strains, and geographical spread due to international livestock exchanges. © The Author(s) 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  14. Molecular characterization of Rhodococcus equi from horse-breeding farms by means of multiplex PCR for the vap gene family.

    PubMed

    Monego, Fernanda; Maboni, Franciele; Krewer, Cristina; Vargas, Agueda; Costa, Mateus; Loreto, Elgion

    2009-04-01

    This study evaluated the molecular characteristics of Rhodococcus equi isolates obtained from horses by a multiplex PCR assay that amplifies the vap gene family (vapA, -B, -C, -D, -E, -F, -G, and -H). A total of 180 R. equi isolates were studied from four different sources, namely healthy horse feces (112), soil (12), stalls (23), and clinical isolates (33) from horse-breeding farms. The technique was performed and confirmed by sequencing of amplified vap gene family controls. Thirty-two (17.8%) of the R. equi isolates were positive for the vapA gene and carried at least three other vap genes. All 147 isolates from equine feces, stalls, and soil failed to demonstrate any genes associated with virulence-inducing proteins. About 32 (97.0%) out of the 33 clinical equine isolates tested positive for the multiplex PCR assay for the vap gene family. They demonstrated six molecular profiles: 100% featured the vapA, vapD, and vapG genes, 86.6% vapF, 76.6% vapH, 43.3% vapC, 36.6% vapE, and none vapB. The most frequent molecular profile was vap A, -D, -F, G, and -H, where this profile was present in 37.5% of the strains. Moreover, there was no molecular epidemiological pattern for R. equi isolates that uniquely mapped to each horse-breeding farm studied. Our proposed technique allows the identification of eight members of the vap gene family (vapA, B, -C, -D, -E, -F, -G, and -H). It is a practical and efficient method of conducting clinical and epidemiological studies on R. equi isolates.

  15. Identification and mutagenesis by allelic exchange of choE, encoding a cholesterol oxidase from the intracellular pathogen Rhodococcus equi.

    PubMed

    Navas, J; González-Zorn, B; Ladrón, N; Garrido, P; Vázquez-Boland, J A

    2001-08-01

    The virulence mechanisms of the facultative intracellular parasite Rhodococcus equi remain largely unknown. Among the candidate virulence factors of this pathogenic actinomycete is a secreted cholesterol oxidase, a putative membrane-damaging toxin. We identified and characterized the gene encoding this enzyme, the choE monocistron. Its protein product, ChoE, is homologous to other secreted cholesterol oxidases identified in Brevibacterium sterolicum and Streptomyces spp. ChoE also exhibits significant similarities to putative cholesterol oxidases encoded by Mycobacterium tuberculosis and Mycobacterium leprae. Genetic tools for use with R. equi are poorly developed. Here we describe the first targeted mutagenesis system available for this bacterium. It is based on a suicide plasmid, a selectable marker (the aacC4 apramycin resistance gene from Salmonella), and homologous recombination. The choE allele was disrupted by insertion of the aacC4 gene, cloned in pUC19 and introduced by electroporation in R. equi. choE recombinants were isolated at frequencies between 10(-2) and 10(-3). Twelve percent of the recombinants were double-crossover choE mutants. The choE mutation was associated with loss of cooperative (CAMP-like) hemolysis with sphingomyelinase-producing bacteria (Listeria ivanovii). Functional complementation was achieved by expression of choE from pVK173-T, a pAL5000 derivative conferring hygromycin resistance. Our data demonstrate that ChoE is an important cytolytic factor for R. equi. The highly efficient targeted mutagenesis procedure that we used to generate choE isogenic mutants will be a valuable tool for the molecular analysis of R. equi virulence.

  16. Re-Emergence of the Apicomplexan Theileria equi in the United States: Elimination of Persistent Infection and Transmission Risk

    PubMed Central

    Ueti, Massaro W.; Mealey, Robert H.; Kappmeyer, Lowell S.; White, Stephen N.; Kumpula-McWhirter, Nancy; Pelzel, Angela M.; Grause, Juanita F.; Bunn, Thomas O.; Schwartz, Andy; Traub-Dargatz, Josie L.; Hendrickson, Amy; Espy, Benjamin; Guthrie, Alan J.; Fowler, W. Kent; Knowles, Donald P.

    2012-01-01

    Arthropod-borne apicomplexan pathogens that cause asymptomatic persistent infections present a significant challenge due to their life-long transmission potential. Although anti-microbials have been used to ameliorate acute disease in animals and humans, chemotherapeutic efficacy for apicomplexan pathogen elimination from a persistently infected host and removal of transmission risk is largely unconfirmed. The recent re-emergence of the apicomplexan Theileria equi in U.S. horses prompted testing whether imidocarb dipropionate was able to eliminate T. equi from naturally infected horses and remove transmission risk. Following imidocarb treatment, levels of T. equi declined from a mean of 104.9 organisms/ml of blood to undetectable by nested PCR in 24 of 25 naturally infected horses. Further, blood transfer from treated horses that became nested PCR negative failed to transmit to naïve splenectomized horses. Although these results were consistent with elimination of infection in 24 of 25 horses, T. equi-specific antibodies persisted in the majority of imidocarb treated horses. Imidocarb treatment was unsuccessful in one horse which remained infected as measured by nested PCR and retained the ability to infect a naïve recipient via intravenous blood transfer. However, a second round of treatment eliminated T. equi infection. These results support the utility of imidocarb chemotherapy for assistance in the control and eradication of this tick-borne pathogen. Successful imidocarb dipropionate treatment of persistently infected horses provides a tool to aid the global equine industry by removing transmission risk associated with infection and facilitating international movement of equids between endemic and non-endemic regions. PMID:22970295

  17. Clinical investigation on Theileria equi and Babesia caballi infections in Italian donkeys.

    PubMed

    Laus, Fulvio; Spaterna, Andrea; Faillace, Vanessa; Veronesi, Fabrizia; Ravagnan, Silvia; Beribé, Francesca; Cerquetella, Matteo; Meligrana, Marina; Tesei, Beniamino

    2015-04-28

    Interest in the welfare and diseases of donkeys is constantly increasing in several countries. Despite this, clinical research into donkeys needs to be in continual development since they show different reactions compared to horses in many conditions, including infectious diseases, and need specific clinical and therapeutic approaches. No reports are currently available on clinical and clinical pathology data regarding donkeys with natural piroplasms infection. Venous blood samples were taken from one hundred and thirty eight donkeys and underwent indirect fluorescent antibody test (IFAT) to detect IgG antibodies against Theileria equi and Babesia caballi and real-time polimerase chain reaction (PCR) to detect Babesia spp. and Theileria spp. Clinical examinations, haematological analyses and serum bilirubin evaluation were also performed and compared with positive or negative status. A seroprevalence of 40.6% and 47.8% was found for T. equi and B. caballi, respectively; double positivity was detected in 19.6% of the animals. PCR results showed that 17.4% of the animals tested positive for T.equi and 3.6% for B. caballi with no double positivity. Twelve donkeys (8.7%) had clinical signs consistent with chronic forms of the disease and no acute forms were detected. Fifty-eight donkeys had haematological and serum bilirubin alterations and 56 (96.6%) of them were IFAT and/or PCR positive. Changes in erythrocyte number, packed cell volume, hemoglobin concentration, mean corpuscular hemoglobin, platelets number and total bilirubin were significantly associated with positive and symptomatic animals. Nonspecific clinical presentation seems to be very common in donkeys and several clinical pathology alterations persist after natural infection. Therefore, apparently healthy donkeys can have masked but severe clinical pathology alterations. Acute forms are very seldom observed in donkeys. Clinical monitoring of chronically infected donkeys is recommended since such animals

  18. VapB type 8 plasmids in Rhodococcus equi isolated from the small intestine of pigs and comparison of selective culture media.

    PubMed

    Lara, G H B; Takai, S; Sasaki, Y; Kakuda, T; Listoni, F J P; Risseti, R M; de Morais, A B C; Ribeiro, M G

    2015-09-01

    The virulence-plasmid profile of Rhodococcus equi strains isolated from Suidae and humans is similar. Recent evidence suggests that the consumption of pork products contaminated with faeces might be a potential source of R. equi infections in humans, mainly to patients with rhodococcosis without history of contact with pigs or pig farms. This study investigated the virulence-associated genes (vapA and vapB) and plasmid profiles of R. equi among the 150 samples of small intestinal content obtained from slaughtered pigs. In addition, all samples were subjected to microbiological culture in conventional sheep blood agar and CAZ-NB, TCP and TVP selective media. A total of 40 (26·7%) of the samples recovered R. equi, with two samples recovering isolates harbouring the VapB type 8 plasmid. Among the 150 pigs sampled herein, CAZ-NB was considered the best selective medium for the isolation of R. equi from faeces. Our results provide evidence that the contamination of slaughtered pig carcasses with pathogenic R. equi might occur through faeces, representing a public health concern. Furthermore, this study is the first description of R. equi strains carrying the VapB plasmid in the gut of pigs. Intermediately virulent (VapB) is a common plasmid-type harboured by R. equi isolated from pigs and humans with AIDS. Curiously, humans with rhodococcosis usually have no history of contact with pigs or pig farms. Virulence-plasmid profile of 40 R. equi isolated among 150 small intestine content samples from pigs revelled two carrying isolates with the VapB type-8 plasmids. Moreover, comparison of three selective culture media shows that CAZ-NB was the best. Our results provide evidence that contamination of slaughtered pig carcasses with pathogenic R. equi might occur through faeces, representing a public health concern. Furthermore, R. equi carrying VapB type-8 plasmids types are described for the first time in the gut of the pig. © 2015 The Society for Applied

  19. Sequence heterogeneity in the 18S rRNA gene within Theileria equi and Babesia caballi from horses in South Africa.

    PubMed

    Bhoora, Raksha; Franssen, Linda; Oosthuizen, Marinda C; Guthrie, Alan J; Zweygarth, Erich; Penzhorn, Barend L; Jongejan, Frans; Collins, Nicola E

    2009-02-05

    A molecular epidemiological survey of the protozoal parasites that cause equine piroplasmosis was conducted using samples collected from horses and zebra from different geographical locations in South Africa. A total of 488 samples were tested for the presence of Theileria equi and/or Babesia caballi using the reverse line blot hybridization assay. Ten percent of the samples hybridized to the Theileria/Babesia genus-specific probe and not to the B. caballi or T. equi species-specific probes, suggesting the presence of a novel species or genotype. The small subunit of rRNA gene (18S; approximately 1600bp) was amplified and sequenced from 33 of these 488 samples. Sequences were compared with published sequences from the public sequence databases. Twelve distinct T. equi and six B. caballi 18S rRNA sequences were identified. Alignments demonstrated extensive sequence variation in the V4 hypervariable region of the 18S rRNA gene within T. equi. Sequence variation was also found in B. caballi 18S rRNA genes, although there was less variation than observed for T. equi. Phylogenetic analysis based on 18S rRNA gene sequences revealed three T. equi clades and two B. caballi clades in South Africa. The extent of sequence heterogeneity detected within T. equi and B. caballi 18S rRNA genes was unexpected since concerted evolution is thought to maintain homogeneity within repeated gene families, including rRNA genes, in eukaryotes. The findings reported here show that careful examination of variants of the 18S rRNA gene of T. equi and B. caballi is required prior to the development of molecular diagnostic tests to detect these parasites in horses. Species-specific probes must be in designed in regions of the gene that are both conserved within and unique to each species.

  20. Role of CD4+, CD8+ and double negative T-cells in the protection of SCID/beige mice against respiratory challenge with Rhodococcus equi.

    PubMed Central

    Ross, T L; Balson, G A; Miners, J S; Smith, G D; Shewen, P E; Prescott, J F; Yager, J A

    1996-01-01

    To evaluate the contributions of T-lymphocyte subsets in pulmonary immunity against Rhodococcus equi, C.B-17 SCID/beige mice were adoptively transferred with splenic lymphocytes from congenic BALB/c mice previously infected with R. equi. Spleen cells were enriched for either CD4+ or CD8+ populations before inoculation, Flow cytometry showed that each enriched population contained less than 0.5% cross contamination. Groups of adoptively transferred SCID/beige mice were sacrificed 6 and 13 d after intranasal infection with R. equi. Bacterial clearance was measured in the lungs, liver and spleen. Lesion development was assessed by gross and histopathological score and the fate of transferred cells assessed by flow cytometry and by immunohistochemistry. SCID/beige mice receiving either CD4+ or CD8+ T-cells were able to clear the infection better than control mice. On d 6 post-infection, bacterial numbers were significantly lower in the lungs of CD4+ transferred mice as compared to CD8+ mice. By d 13, both groups had cleared R. equi from all organs. CD4+ cells were however identified in the lung and spleen of CD8+ recipients at d 13 making conclusions about the role of CD8+ cells in R. equi clearance impossible. By contrast, no significant increases in CD8+ lymphocytes were observed in the organs of CD4+ recipients. All mice developed suppurative bronchopneumonia but lesions were most severe in the CD4+ group. Immunohistochemistry and flow cytometry confirmed that CD4+ and CD8+ cells had migrated to the lungs of adoptively transferred mice. Serum antibody against R, equi was not detected by ELISA in the recipients. SCID/beige mice receiving CD4-CD8- cells were unable to clear R. equi. The study supports the suggestion that CD4+ cells have a central role in R. equi clearance in mice. Images Figure 2. Figure 5. PMID:8809381

  1. Tracing outbreaks of Streptococcus equi infection (strangles) in horses using sequence variation in the seM gene and pulsed-field gel electrophoresis.

    PubMed

    Lindahl, Susanne; Söderlund, Robert; Frosth, Sara; Pringle, John; Båverud, Viveca; Aspán, Anna

    2011-11-21

    Strangles is a serious respiratory disease in horses caused by Streptococcus equi subspecies equi (S. equi). Transmission of the disease occurs by direct contact with an infected horse or contaminated equipment. Genetically, S. equi strains are highly homogenous and differentiation of strains has proven difficult. However, the S. equi M-protein SeM contains a variable N-terminal region and has been proposed as a target gene to distinguish between different strains of S. equi and determine the source of an outbreak. In this study, strains of S. equi (n=60) from 32 strangles outbreaks in Sweden during 1998-2003 and 2008-2009 were genetically characterized by sequencing the SeM protein gene (seM), and by pulsed-field gel electrophoresis (PFGE). Swedish strains belonged to 10 different seM types, of which five have not previously been described. Most were identical or highly similar to allele types from strangles outbreaks in the UK. Outbreaks in 2008/2009 sharing the same seM type were associated by geographic location and/or type of usage of the horses (racing stables). Sequencing of the seM gene generally agreed with pulsed-field gel electrophoresis profiles. Our data suggest that seM sequencing as a epidemiological tool is supported by the agreement between seM and PFGE and that sequencing of the SeM protein gene is more sensitive than PFGE in discriminating strains of S. equi. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. Purulent meningoventriculitis caused by Streptococcus equi subspecies zooepidemicus in a snow leopard (Panthera uncia).

    PubMed

    Yamaguchi, R; Nakamura, S; Hori, H; Kato, Y; Une, Y

    2012-01-01

    Streptococcus equi subspecies zooepidemicus (SEZ) is a zoonotic pathogen that causes respiratory tract infections in man and animals. SEZ infections are very rare in felids. This report describes purulent meningoventriculitis caused by SEZ in an approximately 16-year-old male snow leopard (Panthera uncia). The animal exhibited neurological signs and died 1 month after their onset. On necropsy examination, the surface blood vessels of the brain were swollen and there was an increased volume and turbidity of cerebrospinal fluid (CSF). Microscopically, suppurative inflammation accompanied by gram-positive cocci was observed in the meninges and near the ventricles. SEZ was isolated from the brain tissue and CSF. This is the first report of infection with SEZ in a felid other than a domestic cat. This animal had not had direct contact with horses, but it had been fed horse flesh that may have been the source of infection.

  3. Transmission of Streptococcus equi Subspecies zooepidemicus Infection from Horses to Humans

    PubMed Central

    Lindahl, Susanne B.; Suomala, Päivi; Karhukorpi, Jari; Vuorinen, Sakari; Koivula, Irma; Väisänen, Tia; Pentikäinen, Jaana; Autio, Tiina; Tuuminen, Tamara

    2013-01-01

    Streptococcus equi subspecies zooepidemicus (S. zooepidemicus) is a zoonotic pathogen for persons in contact with horses. In horses, S. zooepidemicus is an opportunistic pathogen, but human infections associated with S. zooepidemicus are often severe. Within 6 months in 2011, 3 unrelated cases of severe, disseminated S. zooepidemicus infection occurred in men working with horses in eastern Finland. To clarify the pathogen’s epidemiology, we describe the clinical features of the infection in 3 patients and compare the S. zooepidemicus isolates from the human cases with S. zooepidemicus isolates from horses. The isolates were analyzed by using pulsed-field gel electrophoresis, multilocus sequence typing, and sequencing of the szP gene. Molecular typing methods showed that human and equine isolates were identical or closely related. These results emphasize that S. zooepidemicus transmitted from horses can lead to severe infections in humans. As leisure and professional equine sports continue to grow, this infection should be recognized as an emerging zoonosis. PMID:23777752

  4. In vitro antimicrobial activity of gallium maltolate against virulent Rhodococcus equi.

    PubMed

    Coleman, Michelle; Kuskie, Kyle; Liu, Mei; Chaffin, Keith; Libal, Melissa; Giguère, Steeve; Bernstein, Lawrence; Cohen, Noah

    2010-11-20

    The objective of this study was to determine the in vitro antimicrobial activity of gallium maltolate (GaM) against Rhodococcus equi. A total of 98 virulent bacterial isolates from equine clinical cases were examined, of which 19 isolates were known to be resistant to macrolides and rifampin. Isolates were cultured with various concentrations of GaM and minimal inhibitory concentration (MIC) values were determined after 24 and 48 h. Both the MIC(50) and the MIC(90) after 24h of growth were 558 ng/mL (8 μM) and after 48 h of growth were 2230 ng/mL (32 μM). There were no apparent differences between MICs of macrolide-resistant and macrolide-susceptible isolates.

  5. Administration of commercial Rhodococcus equi specific hyperimmune plasma results in variable amounts of IgG against pathogenic bacteria in foals.

    PubMed

    Sanz, M G; Oliveira, A F; Page, A; Horohov, D W

    2014-11-15

    Rhodococcus equi is the most common cause of pneumonia in young foals. A vaccine is not available and the use of R equi-specific hyperimmune plasma (HIP) is common. Despite its widespread use, the efficacy of HIP in preventing disease remains controversial. The objectives of this study were (1) to evaluate the virulence associate protein A (VapA)-specific IgG and IgG subclasses in commercially available R equi HIP and (2) to evaluate serum VapA-specific IgG and IgG subclasses in foals following administration of commercial R equi HIP. Three different lots from four commercial R equi HIP were sampled. VapA-specific IgG and IgG subclasses were evaluated in all samples using an ELISA. Serum was collected from newborn foals either after commercial R equi HIP was administered (n=97) or not (n=70). Serum was also collected from each mare. Administration of HIP significantly (P<0.001) increased VapA-specific IgGs in recipient foals, however, there was a marked variation in VapA-specific IgGs in foals receiving the same product. VapA-specific IgGs were significantly different (P<0.001) between products and varied between lots, with coefficients of variation ranging from 17 to 123 per cent. These results may explain previously reported disparities in HIP efficacy. British Veterinary Association.

  6. Recovering data from historical collections: stratigraphic and spatial reconstruction of the outstanding carnivoran record from the Late Pleistocene Equi cave (Apuane Alps, Italy)

    NASA Astrophysics Data System (ADS)

    Ghezzo, Elena; Palchetti, Alessandro; Rook, Lorenzo

    2014-07-01

    Equi Terme is a hamlet located in northern Tuscany, in Apuan Alps regional Park. An outstanding fossil vertebrate collection housed in Florence is the result of excavations in the Equi cave and shelter during the period 1911-1919. This faunal assemblage (associated with Mousterian artefacts) may be correlated with the middle of MIS 3. All of the specimens recovered at Equi early in the last century were collected with attention to their stratigraphical positions. Detailed field annotation for nearly every specimen allowed us to organize them and attempt a stratigraphical and spatial reconstruction of the fossiliferous deposits. We present the results of the study of the spatial and stratigraphic distribution of the carnivoran species in the Equi cave and shelter, and re-evaluate the taphonomic agents of accumulation and the fossil distribution within the stratigraphic record. In particular, we evaluated the distribution of Panthera pardus, which, unusually for Europe, is abundant in the Equi cave assemblage. This analysis highlights the importance of the re-evaluation of historical collections and allows for future comparisons with data from more recent excavations at the Equi site. The analysis also provides an account of the distribution of carnivorans throughout the stratigraphic record. The constant presence and the predominance of leopards and wolves over lions and smaller carnivorans, allow for evaluations of their ethology and may be related to a short period of sediment accumulation.

  7. Development of EMA-2 recombinant antigen based enzyme-linked immunosorbent assay for seroprevalence studies of Theileria equi infection in Indian equine population.

    PubMed

    Kumar, Sanjay; Kumar, Rajender; Gupta, Ashok K; Yadav, Suresh C; Goyal, Sachin K; Khurana, Sandip K; Singh, Raj K

    2013-11-15

    Equine piroplasmosis is a tick-transmitted protozoan disease caused by Theileria equi and/or Babesia caballi. In the present study, we expressed a 53kDa protein from the truncated EMA-2 gene of T. equi (Indian strain) and developed EMA-2ELISA using this expressed protein. This ELISA is able to detect T. equi-specific antibodies in experimentally infected animals as early as 9 days post-infection. The assay developed was validated with the OIE recommended competitive ELISA (cELISA) on 120 serum samples and significant agreement (kappa=0.93) was observed between results of both the ELISAs which indicates suitability of EMA-2ELISA for use in sero-diagnosis. Diagnostic sensitivity and specificity of EMA-2ELISA - as compared with cELISA - were 0.97 and 0.96, respectively. Analysis of 5651 equine serum samples - collected during 2007-2012 from 12 states of India representing eight agro-climatic zones - by EMA-2ELISA revealed 32.65% seroprevalence of T. equi in India. In conclusion, the EMA-2ELISA developed using the T. equi EMA-2 recombinant protein as antigen for detecting T. equi-specific antibodies has good diagnostic potential for sero-epidemiological surveys.

  8. Quadruplex PCR assay for identification of Corynebacterium pseudotuberculosis differentiating biovar Ovis and Equi.

    PubMed

    Almeida, Sintia; Dorneles, Elaine M S; Diniz, Carlos; Abreu, Vinícius; Sousa, Cassiana; Alves, Jorianne; Carneiro, Adriana; Bagano, Priscilla; Spier, Sharon; Barh, Debmalya; Lage, Andrey P; Figueiredo, Henrique; Azevedo, Vasco

    2017-09-25

    Corynebacterium pseudotuberculosis is classified into two biovars, nitrate-negative biovar Ovis which is the etiologic agent of caseous lymphadenitis in small ruminants and nitrate-positive biovar Equi, which causes abscesses and ulcerative lymphangitis in equines. The aim of this study was to develop a quadruplex PCR assay that would allow simultaneous detection and biovar-typing of C. pseudotuberculosis. In the present study, genomes of C. pseudotuberculosis strains were used to identify the genes involved in the nitrate reduction pathway to improve a species identification three-primer multiplex PCR assay. The nitrate reductase gene (narG) was included in the PCR assay along with the 16S, rpoB and pld genes to enhance the diagnosis of the multiplex PCR at biovar level. A novel quadruplex PCR assay for C. pseudotuberculosis species and biovar identification was developed. The results of the quadruplex PCR of 348 strains, 346 previously well-characterized clinical isolates of C. pseudotuberculosis from different hosts (goats, sheep, horse, cattle, buffalo, llamas and humans), the vaccine strain 1002 and the type strain ATCC 19410(T), were compared to the results of nitrate reductase identification by biochemical test. The McNemar's Chi-squared test used to compare the two methods used for C. pseudotuberculosis biovar identification showed no significant difference (P = 0.75) [95% CI for odds ratio (0.16-6.14)] between the quadruplex PCR and the nitrate biochemical test. Concordant results were observed for 97.13% (338 / 348) of the tested strains and the kappa value was 0.94 [95% CI (0.90-0.98)]. The ability of the quadruplex assay to discriminate between C. pseudotuberculosis biovar Ovis and Equi strains enhances its usefulness in the clinical microbiology laboratory.

  9. Sequence heterogeneity in the 18S rRNA gene in Theileria equi from horses presented in Switzerland.

    PubMed

    Liu, Qin; Meli, Marina L; Zhang, Yi; Meili, Theres; Stirn, Martina; Riond, Barbara; Weibel, Beatrice; Hofmann-Lehmann, Regina

    2016-05-15

    A reverse line blot (RLB) hybridization assay was adapted and applied for equine blood samples collected at the animal hospital of the University of Zurich to determine the presence of piroplasms in horses in Switzerland. A total of 100 equine blood samples were included in the study. The V4 hypervariable region of the 18S rRNA gene was amplified by polymerase chain reaction and analyzed using the RLB assay. Samples from seven horses hybridized to a Theileria/Babesia genus-specific and a Theileria genus-specific probe. Of these, two hybridized also to the Theileria equi-specific probe. The other five positive samples did not hybridize to any of the species-specific probes, suggesting the presence of unrecognized Theileria variants or genotypes. The 18S rRNA gene of the latter five samples were sequenced and found to be closely related to T. equi isolated from horses in Spain (AY534822) and China (KF559357) (≥98.4% identity). Four of the seven horses that tested positive had a documented travel history (France, Italy, and Spain) or lived abroad (Hungary). The present study adds new insight into the presence and sequence heterogeneity of T. equi in Switzerland. The results prompt that species-specific probes must be designed in regions of the gene unique to T. equi. Of note, none of the seven positive horses were suspected of having Theileria infection at the time of presentation to the clinic. Clinicians should be aware of the possibility of equine piroplasma infections outside of endemic areas and in horses without signs of piroplasmosis.

  10. Sequential Evaluation of Dogs Naturally Infected with Ehrlichia canis, Ehrlichia chaffeensis, Ehrlichia equi, Ehrlichia ewingii, or Bartonella vinsonii

    PubMed Central

    Breitschwerdt, Edward B.; Hegarty, Barbara C.; Hancock, Susan I.

    1998-01-01

    Historically, disease manifestations in dogs seroreactive to Ehrlichia canis antigens by indirect immunofluorescent antibody testing have been attributed to infection with either E. canis or Ehrlichia ewingii. A 1996 study by Dawson and colleagues provided PCR evidence that healthy dogs from southeastern Virginia could be naturally infected with Ehrlichia chaffeensis. This observation stimulated us to determine which Ehrlichia spp. infected sick dogs that were referred to our hospital from the same region. Based upon PCR amplification with species-specific primers, sick dogs seroreactive to E. canis antigens were determined to be infected with four Ehrlichia species: E. canis, E. chaffeensis, E. equi, and E. ewingii. Coinfection with three Ehrlichia species (E. canis, E. ewingii, and E. equi) was documented for one dog. An additional canine pathogen presumed to be tick transmitted, Bartonella vinsonii subsp. berkhoffii, was identified in 7 of 12 dogs. Importantly, our results indicate that in naturally infected dogs, E. chaffeensis can cause severe disease manifestations that are clinically and serologically indistinguishable from disease manifestations of E. canis or E. ewingii. In addition, our findings support the efficacy of doxycycline for treatment of E. canis, E. equi, and E. ewingii infections but indicate that, based upon the persistence of E. chaffeensis DNA for 1 year following treatment, E. chaffeensis infection in dogs may be more refractory to doxycycline treatment. Undetected coinfection with Bartonella may also complicate the evaluation of treatment efficacy while resulting in disease manifestations that mimic ehrlichiosis. PMID:9705408

  11. A field survey for the seroprevalence of Theileria equi and Babesia caballi in donkeys from Nuu Division, Kenya.

    PubMed

    Oduori, David O; Onyango, Solomon C; Kimari, Joseph N; MacLeod, Ewan T

    2015-07-01

    Equine piroplasmosis is one of the most significant tick-borne disease of equids. The prevalence of this disease in donkeys of semi-arid Kenya remains largely unexplored. The primary objective of this study was to demonstrate the extent to which donkeys in Nuu division, Kenya have been exposed to the haemoprotozoans Babesia caballi and Theileria equi, the causative agents of equine piroplasmosis. The study also assessed the effect of age and sex on seroprevalence. A stratified sampling approach was used and three hundred and fourteen donkeys were sampled across nine sub-locations in Nuu division, Mwingi district. Serodiagnosis was via competitive inhibition enzyme linked immunosorbent assays (cELISA). The seroprevalence of T. equi was 81.2% (95% CI: 76.4-85.4). There was no significant difference in sub-location seropositivity, gender seropositivity or age related seropositivity. Antibodies against B. caballi were not detected (95% CI: 0-1.2). Findings from this study suggest that T. equi infection is endemic in Nuu division, Mwingi where it exists in a state of endemic stability. Existence of the infection should be communicated to animal health practitioners and donkey owning communities in the area.

  12. Treatment of natural infestations of the biting louse (Werneckiella equi) on horses using triflumuron, a benzoylurea derivative insect growth regulator.

    PubMed

    Lowden, Stewart; Gray, Stephen; Dawson, Kim

    2007-09-30

    The horse biting louse (Werneckiella equi) is a common global equine ectoparasite. To our knowledge, benzoyl(phenyl)urea insecticides (triflumuron, diflubenzuron) commonly used as sheep lousicides, have not been evaluated for efficacy against W. equi. The aim of this study was to determine louse control efficacy, general wellness and dermal safety following triflumuron application as a backline pour-on to horses. Two efficacy trials using 25 adult naturally infested lousy horses, and a dermal safety trial using 10 adult louse-free horses were conducted over a 14-month period. Lousy animals were selected by assessment of their lice status prior to treatment. For the efficacy trial, the triflumuron product was applied at a dose of 2.5mg triflumuron per kg bodyweight (1 mL product per 10 kg bodyweight). For the safety study, triflumuron was applied at a 3x clinical dose of 7.5 mg triflumuron per kg bodyweight (3 mL product per 10 kg bodyweight). In our first efficacy trial, 100% lousicidal efficacy was achieved by day 44 post-treatment. In our second trial, no lice were identified on horses by day 71 post-treatment. In the safety trial, no adverse effects were seen. Results of this study demonstrate that the off-label, experimental pour-on application of triflumuron at 2.5 mg/kg bodyweight is convenient, highly effective and safe (at 3x the clinical dose) for the treatment of the horse biting louse, W. equi.

  13. Development of ELISA test for determination of the level of antibodies against Rhodococcus equi in equine serum and colostrum.

    PubMed

    Witkowski, Lucjan; Kaba, Jarosław; Rzewuska, Magdalena; Nowicki, Mariusz; Szaluś-Jordanow, Olga; Kita, Jerzy

    2012-10-15

    Rhodococcus equi infection occurs worldwide and is one of the major causes of losing foals in the first six months of life. The application of serological tests in the diagnostics of rhodococcosis is limited, however they play a crucial role in immunological studies. The objective of this study was to develop and standardize ELISA test for the determination of the level of antibodies against Rhodococcus equi in equine serum and colostrum.Bacterial cell lysate was used as antigen. The test was standardized on 175 sera obtained from adult horses kept on rhodococcosis-free and endemic farms. Positive and negative control sera were used. The test detected IgG antibodies mainly against VapA protein, which was confirmed by Western blot analysis. The test was easy to perform, did not require inactivation of sera and had low well-to-well variation. The shelf life of antigen-coated ELISA plates was 21 days.The test allowed to reveal significant increase of R. equi-specific antibodies in both serum and colostrum in response to the vaccination (p<0.001). Therefore it can be applied to the evaluation of efficacy of immunization. Moreover, no statistically significant difference in the baseline antibody level in adult horses from rhodococcosis-free and endemic farm was revealed (α=0.05). Copyright © 2012 Elsevier B.V. All rights reserved.

  14. Unsaturated fatty acids as modulators of macrophage respiratory burst in the immune response against Rhodococcus equi and Pseudomonas aeruginosa.

    PubMed

    Adolph, Stephanie; Schoeniger, Axel; Fuhrmann, Herbert; Schumann, Julia

    In this paper, using the monocyte/macrophage cell line RAW264.7, we systematically investigate the impact of macrophage enrichment with unsaturated fatty acids on cellular radical synthesis. We found that the intracellular production of reactive nitrogen and oxygen intermediates depends on the activation status of the macrophages. For unstimulated macrophages PUFA enrichment resulted in an increase in cellular radical synthesis. For stimulated macrophages, instead, an impeding action of unsaturated fatty acids on the respiratory burst could be seen. Of particular importance, the impact of unsaturated fatty acids on the macrophage respiratory burst was also observed in RAW264.7 cells cocultivated with viable bacteria of the species Rhodococcus equi or Pseudomonas aeruginosa. PUFA supplementation of macrophages in the presence of R. equi or P. aeruginosa reduced the pathogen-stimulated synthesis of reactive nitrogen and oxygen intermediates. Furthermore, the unsaturated fatty acids were found to impede the expression of the myeloperoxidase gene and to reduce the activity of the enzyme. Hence, our data provide indications of a possible value of PUFA application to people suffering from chronic infections with R. equi and P. aeruginosa as a concomitant treatment to attenuate an excessive respiratory burst. Copyright © 2012 Elsevier Inc. All rights reserved.

  15. Development and evaluation of the internal-controlled real-time PCR assay for Rhodococcus equi detection in various clinical specimens

    PubMed Central

    STEFAŃSKA, Ilona; WITKOWSKI, Lucjan; RZEWUSKA, Magdalena; DZIECIĄTKOWSKI, Tomasz

    2015-01-01

    Rhodococcus equi is the causative agent of rhodococcosis in horses, resulting in significant morbidity and mortality in foals. This bacterium has also been isolated from a variety of animals and is being increasingly reported as a cause of infection in humans, mainly in immunosuppressed individuals. Laboratory diagnostics of R. equi infections based only on conventional microbiological methods shows low accuracy and can lead to misidentification. The objective of the study was to develop and evaluate a real-time PCR assay for direct detection of R. equi in various clinical specimens, including tissue samples. The species-specific region of the gene encoding R. equi cholesterol oxidase, choE, was used as a qPCR-target. The diagnostic applicability of the assay was confirmed by testing various tissue specimens obtained from horses with clinical signs of rhodoccocal infection and swine submaxillary lymph nodes. The rate of R. equi detection in clinical specimens by the developed assay was higher in comparison to the culture method (90% vs. 60.0% of positive samples) and conventional PCR (90.0% vs. 20.0% of positive samples). In case of 13 samples that were negative in the culture-based method, R. equi was detected by the developed assay. Only in one case, it gave negative result for culture-positive sample. The assay may provide a simple and rapid tool to complement the classical methods of R. equi detection based on culture and phenotypic identification of isolates, as the performed evaluation indicated a high specificity and accuracy of the results. PMID:26655770

  16. Development and evaluation of the internal-controlled real-time PCR assay for Rhodococcus equi detection in various clinical specimens.

    PubMed

    Stefańska, Ilona; Witkowski, Lucjan; Rzewuska, Magdalena; Dzieciątkowski, Tomasz

    2016-05-03

    Rhodococcus equi is the causative agent of rhodococcosis in horses, resulting in significant morbidity and mortality in foals. This bacterium has also been isolated from a variety of animals and is being increasingly reported as a cause of infection in humans, mainly in immunosuppressed individuals. Laboratory diagnostics of R. equi infections based only on conventional microbiological methods shows low accuracy and can lead to misidentification. The objective of the study was to develop and evaluate a real-time PCR assay for direct detection of R. equi in various clinical specimens, including tissue samples. The species-specific region of the gene encoding R. equi cholesterol oxidase, choE, was used as a qPCR-target. The diagnostic applicability of the assay was confirmed by testing various tissue specimens obtained from horses with clinical signs of rhodoccocal infection and swine submaxillary lymph nodes. The rate of R. equi detection in clinical specimens by the developed assay was higher in comparison to the culture method (90% vs. 60.0% of positive samples) and conventional PCR (90.0% vs. 20.0% of positive samples). In case of 13 samples that were negative in the culture-based method, R. equi was detected by the developed assay. Only in one case, it gave negative result for culture-positive sample. The assay may provide a simple and rapid tool to complement the classical methods of R. equi detection based on culture and phenotypic identification of isolates, as the performed evaluation indicated a high specificity and accuracy of the results.

  17. Novel description of force of infection and risk factors associated with Theileria equi in horses in Israel and in The Palestinian Authority.

    PubMed

    Aharonson-Raz, Karin; Rapoport, Adi; Hawari, Ibrahim M; Lensky, Itamar M; Berlin, Dalia; Zivotofsky, Doni; Klement, Eyal; Steinman, Amir

    2014-06-01

    The objectives of this study were to determine the force of infection (FOI) for Theileria equi in horses in Israel and to evaluate risk factors associated with seroprevalence a decade after it was last determined by PCR, in 2002. Using a commercial cELISA kit, we demonstrated a widespread and constant exposure to T. equi in Israel (110/216; 50.9%) and the Palestinian Authority (32/108; 29.6%). Owing to the paired samples collected from the same horses approximately one year apart, we were able to determine the FOI rate with which susceptible individuals become infected. Out of the 75 naïve horses in the first collection, four seroconverted during 10-16 months, demonstrating an FOI of 5% for that period. Similar results were obtained by calculating FOI using age-specific seroprevalence (4.2% per year). Housing management type was significantly associated with T. equi seroprevalence with 87.9% seropositivity in horses on pasture and 32.6% seropositivity in horses in stalls/yards. This strong association and the very high seroprevalence found in horses held on pasture, prompted stratification of data accordingly. Geographical location of horses in Israel showed a strong association with seroprevalence to T. equi ranging from 34.5% in central Israel to 80.8% in the northern part of the country. However, when analyzing this association only in horses held in stalls/yards, the lower seroprevalence was noted in the north. In addition, age was significantly associated with seroprevalence for T. equi only in horses held in stalls/yards (R(2)=0.94). Environmental variables were not found to be associated with seroprevalence for T. equi. Here, we report for the first time the FOI for T. equi in horses and highlight the influence housing management type has on the evaluation of risk factors associated with a vector-borne disease, perhaps leading to the discrepancies observed between studies throughout the world.

  18. Production of recombinant EMA-1 protein and its application for the diagnosis of Theileria equi using an enzyme immunoassay in horses from São Paulo State, Brazil.

    PubMed

    Baldani, Cristiane Divan; Hilario, Eduardo; Nakaghi, Andréa Cristina Higa; Bertolini, Maria Célia; Machado, Rosangela Zacarias

    2011-01-01

    The erythrocytic-stage surface protein, Equi Merozoite Antigen 1 (EMA-1), is a major candidate for the development of a diagnostic antigen for equine piroplasmosis. In order to establish an effective diagnostic method for practical use, the gene encoding the entire EMA-1 of Theileria equi Jaboticabal strain was cloned and expressed in Escherichia coli as a histidine-tagged protein (His6-EMA1). The expressed EMA-1 reacted with specific antibodies in Western blot and had an apparent molecular mass of 34 kDa which was largely consistent with its theoretical value. The nucleotide sequence of the EMA-1 gene of Jaboticabal strain was comparatively analyzed with other published sequences. The results indicated a high degree of homology with EMA-1 genes of all other strains isolated from various countries. The recombinant purified His6-EMA1 protein was tested in an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies anti-T. equi in horses. The ELISA clearly differentiated T. equi-infected from Babesia caballi-infected horse sera or normal horse sera. Field serum samples collected from horses in the State of São Paulo, Southeastern Brazil, were examined for the diagnosis of T. equi infection by ELISA. Of 170 samples analyzed, 95.88% (163/170) were positive for T. equi infection. These results suggest that the His6-EMA1 protein expressed in E. coli could be a reliable immunodiagnostic antigen for ELISA test and that T. equi infection is a serious concern in the State of São Paulo, Brazil.

  19. Prevalence of Streptococcus dysgalactiae subsp. equisimilis and S. equi subsp. zooepidemicus in a sample of healthy dogs, cats and horses.

    PubMed

    Acke, E; Midwinter, A C; Lawrence, K; Gordon, S J G; Moore, S; Rasiah, I; Steward, K; French, N; Waller, A

    2015-09-01

    To estimate the prevalence of β-haemolytic Lancefield group C streptococci in healthy dogs, cats and horses; to determine if frequent contact with horses was associated with isolation of these species from dogs and cats; and to characterise recovered S. equi subsp. zooepidemicus isolates by multilocus sequence typing. Oropharyngeal swabs were collected from 197 dogs and 72 cats, and nasopharyngeal swabs from 93 horses. Sampling was carried out at the Massey University Veterinary Teaching Hospital, on sheep and beef farms or on premises where horses were present. All animals were healthy and were categorised as Urban dogs and cats (minimal contact with horses or farm livestock), Farm dogs (minimal contact with horses) and Stable dogs and cats (frequent contact with horses). Swabs were cultured for β-haemolytic Streptococcus spp. and Lancefield group C streptococcal subspecies were confirmed by phenotypic and molecular techniques. Of the 197 dogs sampled, 21 (10.7 (95% CI= 4.0-25.4)%) tested positive for S. dysgalactiae subsp. equisimilis and 4 (2.0 (95% CI=0.7-5.5)%) tested positive for S. equi subsp. zooepidemicus. All these isolates, except for one S. dysgalactiae subsp. equisimilis isolate in an Urban dog, were from Stable dogs. S. dysgalactiae subsp. equisimilis was isolated from one Stable cat. Of the 93 horses, 22 (23.7 (95% CI=12.3-40.6)%) and 6 (6.5 (95% CI=2.8-14.1)%) had confirmed S. dysgalactiae subsp. equisimilis and S. equi subsp. zooepidemicus isolation respectively. Isolation of S. dysgalactiae subsp. equisimilis from dogs was associated with frequent contact with horses (OR=9.8 (95% CI=2.6-72.8)). Three different multilocus sequence type profiles of S. equi subsp. zooepidemicus that have not been previously reported in dogs were recovered. Subclinical infection or colonisation by S. equi subsp. zooepidemicus and S. dysgalactiae subsp. equisimilis occurs in dogs and further research on inter-species transmission and the pathogenic potential of these

  20. [Development and characterization of monoclonal Antibody against M-like protein of Streptococcus equi subsp. zooepidemicus from pig].

    PubMed

    Fan, Hong-jie; He, Shang; Lu, Cheng-ping

    2007-08-01

    Streptococcus equi subsp. zooepidemicus belongs to lancefield group C streptococcus, which can cause disease both in animals and humans. It has been associated with a wide variety of serious infections, including meningitis, pneumonia, septic arthritis and mastitis. The M like proteins on the surface of S. equi subsp. zooepidemicus have an antiphagocytic role analogous to that of group A streptococcal M proteins that are essential in establishing infection. In the present study, the M-like gene without partial signal peptide sequence was amplified from genomic DNA of S. equi subsp. zooepidemicus ATCC35246 strain isolated from pig by polymerase chain reaction (PCR). Then the amplified fragment was cloned in the proper orientation into the site between EcoR I and Xho I of pET32-a(+) via restriction endonuclease EcoR I and Xho I. The recombinant plasmid was verified by restriction endonuclease analysis and nucleotide sequencing, then transformed into E. coli BL21. An fusion protein was expressed in BL21 after induced by IPTG, SDS-PAGE analysis showed that the recombinant protein had a molecular weight of 60 kD, Western blotting showed a positive reaction with the antiserum against ATCC35246. To prepare the monoclonal antibodies (McAbs) against the M-like protein, 6-8 weeks old BABL/c mice were immunized endermicly with purified recombinant M-like protein by Ni-nitrilotriacetic acid affinity chromatography. Splenocytes from the immuniszed mice were fused with SP2/0 and indirect ELISA was used to screen hybridoma cells. 12 hybridoma cell lines secreting McAbs against M-like protein of Streptococcus equi subsp. zooepidemicus were generated, and indirect ELISA confirmed that these McAbs only reacted with M-like protein, but not reacted with other bacteria such as group A Streptococci, Streptococcus suis type 2, Streptococcus equi. The indirect ELISA titers of these 12 ascites McAbs were about 2.56 x 10(4) to 1.01 x 10(5) , and the subtype of these McAbs belong to IgG2b

  1. Vaccination of Mice with Virulence-Associated Protein G (VapG) Antigen Confers Partial Protection against Rhodococcus equi Infection through Induced Humoral Immunity

    PubMed Central

    Trevisani, Marcel M.; Hanna, Ebert S.; Oliveira, Aline F.; Cardoso, Silvia A.; Roque-Barreira, Maria C.; Soares, Sandro G.

    2017-01-01

    Rhodococcus equi is a facultative intracellular bacterium causing severe pyogranulomatous pneumonia, ulcerative enterocolitis, and mesenteric lymphadenopathy in foals aged less than 6 months. Less frequently, this pathogen affects various other species, such as pigs, cattle, cats, and even humans. Although rhodococcosis is treated with a combination of antimicrobial agents, resistance is developed in some cases, and thus, antimicrobial susceptibility must be monitored and managed. Considering these limitations of the current therapy and unavailability of a vaccine to prevent the disease, research is particularly focused on the development of an effective vaccine against rhodococcosis. Most vaccines undergoing development utilize the virulence-associated protein (Vap) A antigen, which was identified previously as a key virulence factor of R. equi. Nevertheless, other proteins, such as VapG, present in most virulent R. equi strains, are also encoded by vap genes located on the R. equi bacterial virulence plasmid. In the present study, we evaluated the effect of VapG immunization on the survival of R. equi-challenged mice. We used attenuated Salmonella as a carrier for VapG (Salmonella-vapG+), a procedure previously adopted to develop a VapA-based vaccine. We observed that vaccination with Salmonella-vapG+ induced both an increased IFN-γ, IL-12, and TNF-α production, and a decreased bacterial burden in organs of the R. equi-challenged mice. Nevertheless, Salmonella-vapG+ vaccination protected only 50% of the mice challenged with a lethal dose of R. equi. Interestingly, we observed an increased frequency of B cells in the spleen of Salmonella-vapG+-vaccinated mice and showed that Salmonella-vapG+-vaccinated R. equi-challenged B-cell-knockout mice did not reduce the bacterial burden. Given these results, we discussed the potential role of the humoral immune response induced by Salmonella-vapG+ vaccination in conferring protection against R. equi infection, as well as

  2. Vaccination of Mice with Virulence-Associated Protein G (VapG) Antigen Confers Partial Protection against Rhodococcus equi Infection through Induced Humoral Immunity.

    PubMed

    Trevisani, Marcel M; Hanna, Ebert S; Oliveira, Aline F; Cardoso, Silvia A; Roque-Barreira, Maria C; Soares, Sandro G

    2017-01-01

    Rhodococcus equi is a facultative intracellular bacterium causing severe pyogranulomatous pneumonia, ulcerative enterocolitis, and mesenteric lymphadenopathy in foals aged less than 6 months. Less frequently, this pathogen affects various other species, such as pigs, cattle, cats, and even humans. Although rhodococcosis is treated with a combination of antimicrobial agents, resistance is developed in some cases, and thus, antimicrobial susceptibility must be monitored and managed. Considering these limitations of the current therapy and unavailability of a vaccine to prevent the disease, research is particularly focused on the development of an effective vaccine against rhodococcosis. Most vaccines undergoing development utilize the virulence-associated protein (Vap) A antigen, which was identified previously as a key virulence factor of R. equi. Nevertheless, other proteins, such as VapG, present in most virulent R. equi strains, are also encoded by vap genes located on the R. equi bacterial virulence plasmid. In the present study, we evaluated the effect of VapG immunization on the survival of R. equi-challenged mice. We used attenuated Salmonella as a carrier for VapG (Salmonella-vapG+), a procedure previously adopted to develop a VapA-based vaccine. We observed that vaccination with Salmonella-vapG+ induced both an increased IFN-γ, IL-12, and TNF-α production, and a decreased bacterial burden in organs of the R. equi-challenged mice. Nevertheless, Salmonella-vapG+ vaccination protected only 50% of the mice challenged with a lethal dose of R. equi. Interestingly, we observed an increased frequency of B cells in the spleen of Salmonella-vapG+-vaccinated mice and showed that Salmonella-vapG+-vaccinated R. equi-challenged B-cell-knockout mice did not reduce the bacterial burden. Given these results, we discussed the potential role of the humoral immune response induced by Salmonella-vapG+ vaccination in conferring protection against R. equi infection, as well as

  3. TRPM2 SNP genotype previously associated with susceptibility to Rhodococcus equi pneumonia in Quarter Horse foals displays differential gene expression identified using RNA-Seq.

    PubMed

    McQueen, Cole M; Whitfield-Cargile, Canaan M; Konganti, Kranti; Blodgett, Glenn P; Dindot, Scott V; Cohen, Noah D

    2016-12-05

    Rhodococcus equi (R. equi) is an intracellular bacterium that affects young foals and immuno-compromised individuals causing severe pneumonia. Currently, the genetic mechanisms that confer susceptibility and/or resistance to R. equi are not fully understood. Previously, using a SNP-based genome-wide association study, we identified a region on equine chromosome 26 associated with culture-confirmed clinical pneumonia. To better characterize this region and understand the function of the SNP located within TRPM2 that was associated with R. equi pneumonia, we performed RNA-Seq on 12 horses representing the 3 genotypic forms of this SNP. We identified differentially expressed genes in the innate immune response pathway when comparing homozygous A allele horses with the AB and BB horses. Isoform analyses of the RNA-Seq data predicted the existence of multiple transcripts and provided evidence of differential expression at the TRPM2 locus. This finding is consistent with previously demonstrated work in human cell lines in which isoform-specific expression of TRPM2 was critical for cell viability. This work demonstrates that SNPs in TRPM2 are associated with differences in gene expression, suggesting that modulation of expression of this innate immune gene contributes to susceptibility to R. equi pneumonia.

  4. Evaluation of equine breeding farm management and preventative health practices as risk factors for development of Rhodococcus equi pneumonia in foals.

    PubMed

    Chaffin, M Keith; Cohen, Noah D; Martens, Ronald J

    2003-02-15

    To determine whether foal management practices, environmental management, and preventative health practices are risk factors for development of Rhodococcus equi pneumonia in foals. Prospective matched case-control study. 2,764 foals on 64 equine breeding farms with 9,991 horses. During 1997, participating veterinarians completed paired data collection forms for comparison; 1 for an affected farm (containing > or = 1 foal with pneumonia caused by R equi) and 1 for a control farm. Information collected pertained to stabling facilities, environmental management, foal husbandry, and preventative equine health practices. Matched farm data compared by use of conditional logistic regression indicated that personnel on affected farms were more likely to attend foal births, test foals for adequacy of passive immunity, administer plasma or other treatments to foals to supplement serum immunoglobulin concentrations, administer hyperimmune plasma prophylactically to foals, vaccinate mares and foals against Streptococcus equi infection, and use multiple anthelmintics in deworming programs. Affected farms were also more likely to have foals that developed other respiratory tract disorders and were approximately 4 times as likely to have dirt floors in stalls used for housing foals as were control farms. Rhodococcus equi pneumonia does not appear to be associated with poor farm management or a lack of attention to preventative health practices. Housing foals in stalls with dirt floors may increase the risk for development of R equi pneumonia.

  5. Genome and proteome analysis of phage E3 infecting the soil-borne actinomycete Rhodococcus equi.

    PubMed

    Salifu, Samson P; Valero-Rello, Ana; Campbell, Samantha A; Inglis, Neil F; Scortti, Mariela; Foley, Sophie; Vázquez-Boland, José A

    2013-02-01

    We report on the characterization and genomic analysis of bacteriophage E3 isolated from soil and propagating in Rhodococcus equi strains. Phage E3 has a circular genome of 142 563 bp and is the first Myoviridae reported for the genus Rhodococcus and for a non-mycobacterial actinomycete. Phylogenetic analyses placed E3 in a distinct Myoviridae clade together with Mycobacterium phages Bxz1 and Myrna. The highly syntenic genomes of this myoviridal group comprise vertically evolving core phage modules flanked by hyperplastic regions specific to each phage and rich in horizontally acquired DNA. The hyperplastic regions contain numerous tRNA genes in the mycobacteriophages which are absent in E3, possibly reflecting bacterial host-specific translation-related phage fitness constraints associated with rate-limiting tRNAs. A structural proteome analysis identified 28 E3 polypeptides, including 15 not previously known to be virion-associated proteins. The E3 genome and comparative analysis provide insight into short-term genome evolution and adaptive plasticity in tailed phages from the environmental microbiome. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  6. Characteristics of wettedness and equi-skin temperature line in the evaporative regulation region

    NASA Astrophysics Data System (ADS)

    Mochida, T.

    1983-07-01

    As a result of the analysis of physiological experimental data, the characteristics of the wettedness were clarified, i.e., the value of the wettedness is not constant but differs in accordance with the environmental humidity even when the skin temperature is the same, and it was shown that the evaporative heat loss from the skin surface is inversely proportional to the wetttedness. Based on the properties of the wetedness observed, a new thermal sensation chart in the evaporative regulation region was proposed as an index for evaluating the warmth or the coldness in the environment. The feature of the present chart is that the locus of the equal skin temperature appears as a curved line on the psychrometric chart and that the wettedness on the equi-skin temperature line is not constant but takes varying values. The curved equal skin temperature line means that the influence of the environmental humidity on thermal sensation becomes smaller as the humidity of the environmental humidity on thermal sensation becomes smaller as the humidity of the environment is lowered.

  7. Spall properties of Al 5083 plate fabricated using equi-channel angular pressing (ECAP) and rolling

    NASA Astrophysics Data System (ADS)

    Whelchel, R. L.; Thadhani, N. N.; Sanders, T. H.; Kecskes, L. J.; Williams, C. L.

    2014-05-01

    The spall strength and Hugoniot Elastic Limit (HEL) of aluminum alloy 5083 (Al 5083) are compared for plates fabricated using equi-channel angular pressing (ECAP) and rolling. Al 5083 is a light-weight and strain-hardenable aluminum alloy used for armor plating in military transport vehicles, thus requiring the highest achievable spall strength and HEL. Materials that were processed by ECAP displayed a highly refined grain structure with little texture and a large degree of plastic deformation, whereas subsequent rolling resulted in a textured microstructure with both grains and inclusions aligning along the rolling direction. The spall behavior of Al 5083 was determined using plate-impact gas-gun experiments with rear free surface velocity measurements for a variety of processing conditions involving both ECAP and rolling. The spall strength and HEL increased from that of the as-received material after processing with ECAP. Subsequent rolling further increased the HEL but reduced the spall strength. Rolling also resulted in directional dependence of the spall strength, with the lowest spall strength occurring for impact through the plate thickness and highest spall strength in the rolling direction. The trends in the spall behavior correlate with the size and preferential alignment of manganese dispersoids and iron and silicon rich inclusions that are evolved during processing.

  8. Identification of novel genes expressed during host infection in Streptococcus equi ssp. zooepidemicus ATCC35246.

    PubMed

    Ma, Zhe; Yu, Lei; Zhou, Hong; Liu, Tingting; Xu, Bin; Ma, Fang; Peng, Jie; Fan, Hongjie

    2015-02-01

    Infection with Streptococcus equi ssp. zooepidemicus (Streptococcus zooepidemicus, SEZ) can cause septicemia, meningitis, and mastitis in domesticated species. Identification of this organism's virulence factors is an effective way of clarifying its pathogenic mechanism. We employed in vivo-induced antigen technology (IVIAT) to find bacterial genes that were only expressed or upregulated in an infected host (IVI genes). Convalescent-phase sera from pigs infected with SEZ were pooled, adsorbed against in vitro antigens, and used to screen SEZ genomic expression libraries. This analysis identified 43 genes as IVI genes. Six of these 43 genes were verified via real-time PCR. Following the analysis, we were able to assign a putative function to 36 of the 43 proteins. These proteins included those involved in virulence and adaptation; formation of intermediary products; gene replication, transcription and expression; energy metabolism; transport and also various proteins of unknown function. The relationship between sagD gene and bacterial virulence was confirmed. This study provides new molecular data for the study of streptococcal disease in swine and is important for identifying the pathogenic mechanisms of SEZ.

  9. Design of a Novel Equi-Biaxial Stretcher for Live Cellular and Subcellular Imaging.

    PubMed

    Imsirovic, Jasmin; Wellman, Tyler J; Mondoñedo, Jarred R; Bartolák-Suki, Elizabeth; Suki, Béla

    2015-01-01

    Cells in the body experience various mechanical stimuli that are often essential to proper cell function. In order to study the effects of mechanical stretch on cell function, several devices have been built to deliver cyclic stretch to cells; however, they are generally not practical for live cell imaging. We introduce a novel device that allows for live cell imaging, using either an upright or inverted microscope, during the delivery of cyclic stretch, which can vary in amplitude and frequency. The device delivers equi-biaxial strain to cells seeded on an elastic membrane via indentation of the membrane. Membrane area strain was calibrated to indenter depth and the device showed repeatable and accurate delivery of strain at the scale of individual cells. At the whole cell level, changes in intracellular calcium were measured at different membrane area strains, and showed an amplitude-dependent response. At the subcellular level, the mitochondrial network was imaged at increasing membrane area strains to demonstrate that stretch can lead to mitochondrial fission in lung fibroblasts. The device is a useful tool for studying transient as well as long-term mechanotransduction as it allows for simultaneous stretching and imaging of live cells in the presence of various chemical stimuli.

  10. Design of a Novel Equi-Biaxial Stretcher for Live Cellular and Subcellular Imaging

    PubMed Central

    Mondoñedo, Jarred R.; Bartolák-Suki, Elizabeth; Suki, Béla

    2015-01-01

    Cells in the body experience various mechanical stimuli that are often essential to proper cell function. In order to study the effects of mechanical stretch on cell function, several devices have been built to deliver cyclic stretch to cells; however, they are generally not practical for live cell imaging. We introduce a novel device that allows for live cell imaging, using either an upright or inverted microscope, during the delivery of cyclic stretch, which can vary in amplitude and frequency. The device delivers equi-biaxial strain to cells seeded on an elastic membrane via indentation of the membrane. Membrane area strain was calibrated to indenter depth and the device showed repeatable and accurate delivery of strain at the scale of individual cells. At the whole cell level, changes in intracellular calcium were measured at different membrane area strains, and showed an amplitude-dependent response. At the subcellular level, the mitochondrial network was imaged at increasing membrane area strains to demonstrate that stretch can lead to mitochondrial fission in lung fibroblasts. The device is a useful tool for studying transient as well as long-term mechanotransduction as it allows for simultaneous stretching and imaging of live cells in the presence of various chemical stimuli. PMID:26466363

  11. Comparison of the Vitek MS and Bruker Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry Systems for Identification of Rhodococcus equi and Dietzia spp.

    PubMed

    de Alegría Puig, Carlos Ruiz; Pilares, Lilian; Marco, Francesc; Vila, Jordi; Martínez-Martínez, Luis; Navas, Jesús

    2017-07-01

    Rhodococcus equi causes pyogranulomatous pneumonia in domesticated animals and immunocompromised humans. Dietzia spp. are environmental bacteria that have rarely been associated with human infections. R. equi and Dietzia spp. are closely related actinomycetes. Phenotypic discrimination between R. equi and Dietzia on the basis of their Gram stain morphology and colony appearance is problematic. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is a fast, reliable, and cost-effective method for identification of a wide variety of microorganisms. We have evaluated the performance of Bruker Biotyper versus that of Vitek MS for identification of a collection of 154 isolates identified at the source as R. equi that includes isolates belonging to the genus Dietzia PCR amplification of the choE gene, encoding a cholesterol oxidase, and 16S rRNA sequencing were considered the reference methods for R. equi identification. Biotyper identified 131 (85.1%) of the 154 isolates at the species level, and this figure increased to 152 (98.7%) when the species cutoff was reduced from a score of ≥2.000 to ≥1.750. Vitek MS correctly identified at the species level 130 (84.4%) isolates as long as bacteria were extracted with ethanol but only 35 (22.7%) isolates when samples were prepared by direct extraction from colonies. The two systems allowed differentiation between R. equi and Dietzia spp., but identification of all Dietzia sp. isolates at the species level needed sequencing of the 16S rRNA gene. Copyright © 2017 American Society for Microbiology.

  12. Molecular characterization of Rhodococcus equi isolates from horses in Poland: pVapA characteristics and plasmid new variant, 85-kb type V.

    PubMed

    Witkowski, Lucjan; Rzewuska, Magdalena; Takai, Shinji; Chrobak-Chmiel, Dorota; Kizerwetter-Świda, Magdalena; Feret, Małgorzata; Gawryś, Marta; Witkowski, Maciej; Kita, Jerzy

    2017-01-26

    Rhodococcus equi is one of the most significant bacterial pathogens affecting foals up to 6 months of age worldwide. Rhodococcosis is present in Poland however information about molecular characterization of R. equi isolates is scarce. This study describes molecular characterization of Rhodococcus equi infection on 13 horse breeding farms in Poland between 2001 and 2012. Samples were collected by tracheobronchial aspiration from pneumonic foals or during necropsy. The R. equi isolates were genotyped by plasmid profiling and pulsed-field gel electrophoresis. Totally, 58 R. equi isolates were investigated. One isolate lost its plasmid. Among the 57 VapA-positive isolates, 48 contained 85-kb type I plasmid (82.8%), 8 contained 87-kb type I plasmid (13.8%). One isolate (1.7%) had a unique restriction cleavage pattern and the 2nd fragment of EcoRI digests of this plasmid DNA was about 2600 bases smaller than that of the 85 kb type I. This new plasmid variant was designated as the "85-kb type V". Among the 58 isolates typeable with VspI-PFGE, ten PFGE clusters were detected. The majority of foals were infected mostly with isolates of low genetic diversity. Most of clinical isolates of R. equi from foals in Poland contain pVapA 85-kb type I and 87-kb type I similarly to the other European countries and the United States. However, the new variant of pVapA 85-kb type V was identified. The chromosomal variability was detected among some of the investigated isolates and the presence of farm-specific isolates might be possible.

  13. Inhibition of the in vitro growth of Babesia bigemina, Babesia caballi and Theileria equi parasites by trifluralin analogues.

    PubMed

    G Silva, Marta; Knowles, Donald P; Antunes, Sandra; Domingos, Ana; Esteves, Maria A; Suarez, Carlos E

    2017-06-01

    Bovine and equine babesiosis caused by Babesia bovis, Babesia bigemina and Babesia caballi, along with equine theileriosis caused by Theileria equi are global tick-borne hemoprotozoan diseases characterized by fever, anemia, weight losses and abortions. A common feature of these diseases are transition from acute to chronic phases, in which parasites may persist in the hosts for life. Antiprotozoal drugs are important for managing infection and disease. Previous research demonstrated that trifluralin analogues, designated (TFLAs) 1-15, which specifically bind to regions of alpha-tubulin protein in plants and protozoan parasites, have the ability to inhibit the in vitro growth of B. bovis. The inhibitory activity of TFLAs 1-15 minus TFLA 5 was tested in vitro against cultured B. bigemina, B. caballi and T. equi. The four TFLAs with greatest inhibitory activity were then analyzed for hemolytic activity and toxicity against erythrocytes. All TFLAs tested in the study showed inhibitory effects against the three parasite species. TFLA 2, TFLA 11, TFLA 13 and TFLA 14 were the most effective inhibitors for the three species tested, with estimated IC50 between 5.1 and 10.1μM at 72h. The drug's solvent (DMSO/ethanol) did not statistically affect the growth of the parasites nor cause hemolysis. Also, TFLA 2, 13 and 14 did not cause statistically significant hemolytic activity on bovine and equine erythrocytes at 15μM, and TFLA 2, 11 and 13 had no detectable toxic effects on bovine and equine erythrocytes at 15μM, suggesting that these drugs do not compromise erythrocyte viability. The demonstrated ability of the trifluralin analogues to inhibit in vitro growth of Babesia spp. and Theileria equi, and their lack of toxic effects on erythrocytes supports further in vivo testing and eventually their development as novel alternatives for the treatment of babesiosis and theileriosis. Copyright © 2017 The Authors. Published by Elsevier GmbH.. All rights reserved.

  14. In-vivo therapeutic efficacy trial with artemisinin derivative, buparvaquone and imidocarb dipropionate against Babesia equi infection in donkeys.

    PubMed

    Kumar, Sanjay; Gupta, Ashok K; Pal, Yash; Dwivedi, Shailendra K

    2003-11-01

    The therapeutic efficacy of imidocarb, artesunate, arteether, buparvaquone and arteether+buparvaquone combination was evaluated against Babesia equi of Indian origin in splenectomised donkeys with experimentally induced acute infection. Efficacies of these drugs were tested by administering each drug or drug combination to groups of donkeys (having three donkeys each group). One group of donkey was kept as untreated control for comparing the results. Parasitaemia, haematology (WBC, RBC, PCV, granulocytes and haemoglobin), biochemical parameters (SAST, SALT, alkaline phosphatase, albumin/globulin ratio) were monitored at regular intervals. Individually, arteether and buparvaquone were found to have no parasite clearing efficacy and the treated animals died within 5-6 days after showing high parasitaemia and clinical symptoms of the disease. However, artesunate treated animals were able to restrict the parasite multiplication but only during the treatment period. Animals treated with imidocarb and arteether+buparvaquone combination were able to clear the parasite from the blood circulation after 2-5 days post-treatment (PT). After 55-58 days PT, recrudescence of B. equi parasite was observed in both these groups and a mean survival period of 66 days and 69 days, respectively, was recorded in these groups. Results of haemato-biochemical parameters had shown that imidocarb had deleterious effect on the liver function while on the other hand arteether+buparvaquone combination was found to be safe. This limited study indicates that arteether+buparvaquone combination could be a better choice than imidocarb for treating B. equi infection, but further trials are required in detail.

  15. Aortic graft infection and mycotic aneurysm with Streptococcus equi zooepidemicus: two cases with favorable outcome of antibiotic treatment.

    PubMed

    Altreuther, Martin; Lange, Conrad; Myhre, Hans Olav; Hannula, Raisa

    2013-02-01

    Infections with Streptococcus equi zooepidemicus are rare and are associated with contact with animals or animal products. There are very few reports about infected vascular grafts or aneurysms with this etiology. We present two patients. The first is a 77-year-old man with an infected bifurcated graft four years after an open operation for an abdominal aortic aneurysm (AAA). The second is a 72-year-old man with a symptomatic mycotic AAA, treated with endovascular aneurysm repair. Both received prolonged treatment with bactericidal antibiotics and responded well. Follow-up time at present is 5.5 years for the first, and 4.5 years for the second, patient.

  16. Haemorrhagic pneumonia in sled dogs caused by Streptococcus equi subsp. zooepidemicus - one fatality and two full recoveries: a case report

    PubMed Central

    2013-01-01

    In spite of yearly vaccination, outbreaks of canine infectious respiratory disease are periodically seen amongst domestic dogs. These infections compromise host defense mechanisms, and, when combined with other stressful events, allow opportunistic pathogens like Streptococcus equi subsp. zooepidemicus to create serious disease. Early recognition and treatment are tremendously important for a successful outcome in these cases. A polyvalent vaccine was given to 22 racing dogs three days after a competition, followed by two days of rest, and then the dogs were returned to regular training. Coughing was noticed among the dogs four days after immunisation. Three days after this outbreak one of the dogs was unusually silent and was found dead the next morning. Simultaneously two other dogs developed haemorrhagic expectorate, depression and dyspnea and were brought in to the veterinary hospital. Streptococcus equi subsp. zooepidemicus was isolated in pure culture from all three cases. They were treated and rehabilitated successfully, and won a sledge race three months later. This paper discusses the necropsy results, treatment regime, rehabilitation and the chronology of vaccination, stressful events and disease. PMID:24020788

  17. The Effect of Deformation Heating on Restoration and Constitutive Equation of a Wrought Equi-Atomic NiTi Alloy

    NASA Astrophysics Data System (ADS)

    Ahadi, A.; Karimi Taheri, A.; Karimi Taheri, K.; Sarraf, I. S.; Abbasi, S. M.

    2012-04-01

    In this study, a set of constitutive equation corrected for deformation heating is proposed for a near equi-atomic NiTi shape memory alloy using isothermal hot compression tests in temperature range of 700 to 1000 °C and strain rate of 0.001 to 1 s-1. In order to determine the temperature rise due to deformation heating, Abaqus simulation was employed and varied thermal properties were considered in the simulation. The results of hot compression tests showed that at low pre-set temperatures and high strain rates the flow curves exhibit a softening, while after correction of deformation heating the softening is vanished. Using the corrected flow curves, the power-law constitutive equation of the alloy was established and the variation of constitutive constants with strain was determined. Moreover, it was found that deformation heating introduces an average relative error of about 9.5% at temperature of 800 °C and strain rate of 0.1 s-1. The very good agreement between the fitted flow stress (by constitutive equation) and the measured ones indicates the accuracy of the constitutive equation in analyzing the hot deformation behavior of equi-atomic NiTi alloy.

  18. Ultrastructural description of a new chytrid genus of caecum anaerobe, Caecomyces equi gen. nov., sp. nov., assigned to the Neocallimasticaceae.

    PubMed

    Gold, J J; Heath, I B; Bauchop, T

    1988-01-01

    Vegetative and reproductive stages of Caecomyces equi gen. nov., sp. nov. isolated from the horse caecum were examined by light and electron microscopy. This organism, which is similar to isolates known as Sphaeromonas communis, produces uniflagellate, uninucleate zoospores whose perikinetosomal structures, i.e. circumflagellar ring, spur, struts and scoop, are similar in many respects to those described in species of Neocallimastix. Microtubular roots extend basally from the spur and associate with hydrogenosomes and the nucleus. Another group of microtubules radiates laterally in a fan-shaped array close to the plasmalemma. Zoospores encyst, shedding their flagella with basal bodies, and germinate to diglobular thalli. Either coralloid or bulbous rhizoids form in plant material, but only the latter in axenic culture. Incipient zoospores are produced from a multinucleate eucarpic thallus and devlop within cleavage vacuoles containing flagella. An isolate from the cow rumen was found to be similar to C. equi in morphology and zoospore ultrastructure. On the basis of zoospore ultrastructure, we assign the new genus to the Neocallimasticaceae of the order Spizellomycetales. Organisms previously described as Sphaeromonas communis and Piromonas communis are renamed Caecomyces communis and Piromyces communis and assigned to the same family.

  19. Differences in Rhodococcus equi Infections Based on Immune Status and Antibiotic Susceptibility of Clinical Isolates in a Case Series of 12 Patients and Cases in the Literature

    PubMed Central

    Suzuki, Yasuhiro; Ribes, Julie A.; Thornton, Alice

    2016-01-01

    Rhodococcus equi is an unusual zoonotic pathogen that can cause life-threatening diseases in susceptible hosts. Twelve patients with R. equi infection in Kentucky were compared to 137 cases reported in the literature. Although lungs were the primary sites of infection in immunocompromised patients, extrapulmonary involvement only was more common in immunocompetent patients (P < 0.0001). Mortality in R. equi-infected HIV patients was lower in the HAART era (8%) than in pre-HAART era (56%) (P < 0.0001), suggesting that HAART improves prognosis in these patients. Most (85–100%) of clinical isolates were susceptible to vancomycin, clarithromycin, rifampin, aminoglycosides, ciprofloxacin, and imipenem. Interestingly, there was a marked difference in susceptibility of the isolates to cotrimoxazole between Europe (35/76) and the US (15/15) (P < 0.0001). Empiric treatment of R. equi infection should include a combination of two antibiotics, preferably selected from vancomycin, imipenem, clarithromycin/azithromycin, ciprofloxacin, rifampin, or cotrimoxazole. Local antibiograms should be checked prior to using cotrimoxazole due to developing resistance. PMID:27631004

  20. Differences in Rhodococcus equi Infections Based on Immune Status and Antibiotic Susceptibility of Clinical Isolates in a Case Series of 12 Patients and Cases in the Literature.

    PubMed

    Gundelly, Praveen; Suzuki, Yasuhiro; Ribes, Julie A; Thornton, Alice

    2016-01-01

    Rhodococcus equi is an unusual zoonotic pathogen that can cause life-threatening diseases in susceptible hosts. Twelve patients with R. equi infection in Kentucky were compared to 137 cases reported in the literature. Although lungs were the primary sites of infection in immunocompromised patients, extrapulmonary involvement only was more common in immunocompetent patients (P < 0.0001). Mortality in R. equi-infected HIV patients was lower in the HAART era (8%) than in pre-HAART era (56%) (P < 0.0001), suggesting that HAART improves prognosis in these patients. Most (85-100%) of clinical isolates were susceptible to vancomycin, clarithromycin, rifampin, aminoglycosides, ciprofloxacin, and imipenem. Interestingly, there was a marked difference in susceptibility of the isolates to cotrimoxazole between Europe (35/76) and the US (15/15) (P < 0.0001). Empiric treatment of R. equi infection should include a combination of two antibiotics, preferably selected from vancomycin, imipenem, clarithromycin/azithromycin, ciprofloxacin, rifampin, or cotrimoxazole. Local antibiograms should be checked prior to using cotrimoxazole due to developing resistance.

  1. Composition and Diversity of the Fecal Microbiome and Inferred Fecal Metagenome Does Not Predict Subsequent Pneumonia Caused by Rhodococcus equi in Foals.

    PubMed

    Whitfield-Cargile, Canaan M; Cohen, Noah D; Suchodolski, Jan; Chaffin, M Keith; McQueen, Cole M; Arnold, Carolyn E; Dowd, Scot E; Blodgett, Glenn P

    2015-01-01

    In equids, susceptibility to disease caused by Rhodococcus equi occurs almost exclusively in foals. This distribution might be attributable to the age-dependent maturation of immunity following birth undergone by mammalian neonates that renders them especially susceptible to infectious diseases. Expansion and diversification of the neonatal microbiome contribute to development of immunity in the gut. Moreover, diminished diversity of the gastrointestinal microbiome has been associated with risk of infections and immune dysregulation. We thus hypothesized that varying composition or reduced diversity of the intestinal microbiome of neonatal foals would contribute to increased susceptibility of their developing R. equi pneumonia. The composition and diversity indices of the fecal microbiota at 3 and 5 weeks of age were compared among 3 groups of foals: 1) foals that subsequently developed R. equi pneumonia after sampling; 2) foals that subsequently developed ultrasonographic evidence of pulmonary abscess formation or consolidation but not clinical signs (subclinical group); and, 3) foals that developed neither clinical signs nor ultrasonographic evidence of pulmonary abscess formation or consolidation. No significant differences were found among groups at either sampling time, indicating absence of evidence of an influence of composition or diversity of the fecal microbiome, or predicted fecal metagenome, on susceptibility to subsequent R. equi pneumonia. A marked and significant difference identified between a relatively short interval of time appeared to reflect ongoing adaptation to transition from a milk diet to a diet including available forage (including hay) and access to concentrate fed to the mare.

  2. Experimental Rhodococcus equi and equine infectious anemia virus DNA vaccination in adult and neonatal horses: Effect of IL-12, dose, and route

    PubMed Central

    Mealey, R.H.; Stone, D.M.; Hines, M.T.; Alperin, D.C.; Littke, M.H.; Leib, S.R.; Leach, S.E.; Hines, S.A.

    2012-01-01

    Improving the ability of DNA-based vaccines to induce potent Type1/Th1 responses against intracellular pathogens in large outbred species is essential. Rhodoccocus equi and equine infectious anemia virus (EIAV) are two naturally occurring equine pathogens that also serve as important large animal models of neonatal immunity and lentiviral immune control. Neonates present a unique challenge for immunization due to their diminished immunologic capabilities and apparent Th2 bias. In an effort to augment R. equi- and EIAV-specific Th1 responses induced by DNA vaccination, we hypothesized that a dual promoter plasmid encoding recombinant equine IL-12 (rEqIL-12) would function as a molecular adjuvant. In adult horses, DNA vaccines induced R. equi- and EIAV-specific antibody and lymphoproliferative responses, and EIAV-specific CTL and tetramer-positive CD8+ T lymphocytes. These responses were not enhanced by the rEqIL-12 plasmid. In neonatal foals, DNA immunization induced EIAV-specific antibody and lymphoproliferative responses, but not CTL. The R. equi vapA vaccine was poorly immunogenic in foals even when co-administered with the IL-12 plasmid. It was concluded that DNA immunization was capable of inducing Th1 responses in horses; dose and route were significant variables, but rEqIL-12 was not an effective molecular adjuvant. Additional work is needed to optimize DNA vaccine-induced Th1 responses in horses, especially in neonates. PMID:17889970

  3. Lymphocytes and macrophages are infected by theileria equi, but T cells and B cells are not required to establish infection in vivo

    USDA-ARS?s Scientific Manuscript database

    Theileria equi has a biphasic life cycle in horses, with a period of intraleukocyte development followed by patent erythrocytic parasitemia that causes acute and sometimes fatal hemolytic disease. Unlike Theileria spp. that infect cattle (Theileria parva and Theileria annulata), the intraleukocyte s...

  4. Composition and Diversity of the Fecal Microbiome and Inferred Fecal Metagenome Does Not Predict Subsequent Pneumonia Caused by Rhodococcus equi in Foals

    PubMed Central

    Whitfield-Cargile, Canaan M.; Cohen, Noah D.; Suchodolski, Jan; Chaffin, M. Keith; McQueen, Cole M.; Arnold, Carolyn E.; Dowd, Scot E.; Blodgett, Glenn P.

    2015-01-01

    In equids, susceptibility to disease caused by Rhodococcus equi occurs almost exclusively in foals. This distribution might be attributable to the age-dependent maturation of immunity following birth undergone by mammalian neonates that renders them especially susceptible to infectious diseases. Expansion and diversification of the neonatal microbiome contribute to development of immunity in the gut. Moreover, diminished diversity of the gastrointestinal microbiome has been associated with risk of infections and immune dysregulation. We thus hypothesized that varying composition or reduced diversity of the intestinal microbiome of neonatal foals would contribute to increased susceptibility of their developing R. equi pneumonia. The composition and diversity indices of the fecal microbiota at 3 and 5 weeks of age were compared among 3 groups of foals: 1) foals that subsequently developed R. equi pneumonia after sampling; 2) foals that subsequently developed ultrasonographic evidence of pulmonary abscess formation or consolidation but not clinical signs (subclinical group); and, 3) foals that developed neither clinical signs nor ultrasonographic evidence of pulmonary abscess formation or consolidation. No significant differences were found among groups at either sampling time, indicating absence of evidence of an influence of composition or diversity of the fecal microbiome, or predicted fecal metagenome, on susceptibility to subsequent R. equi pneumonia. A marked and significant difference identified between a relatively short interval of time appeared to reflect ongoing adaptation to transition from a milk diet to a diet including available forage (including hay) and access to concentrate fed to the mare. PMID:26305682

  5. Comparative bioinformatics analysis of transcription factor genes suggests conservation of key regulatory domains among babesia bovis, B. microti and theileria equi.

    USDA-ARS?s Scientific Manuscript database

    Apicomplexa tick borne hemoparasites including B. bovis, B. microti, and Theileria equi are responsible for bovine and human babesiosis and equine theileriosis respectively. These neglected parasites of vast medical, epidemiological, and economic impact have complex life cycles in their vertebrate a...

  6. Experimental Rhodococcus equi and equine infectious anemia virus DNA vaccination in adult and neonatal horses: effect of IL-12, dose, and route.

    PubMed

    Mealey, R H; Stone, D M; Hines, M T; Alperin, D C; Littke, M H; Leib, S R; Leach, S E; Hines, S A

    2007-10-23

    Improving the ability of DNA-based vaccines to induce potent Type1/Th1 responses against intracellular pathogens in large outbred species is essential. Rhodoccocus equi and equine infectious anemia virus (EIAV) are two naturally occurring equine pathogens that also serve as important large animal models of neonatal immunity and lentiviral immune control. Neonates present a unique challenge for immunization due to their diminished immunologic capabilities and apparent Th2 bias. In an effort to augment R. equi- and EIAV-specific Th1 responses induced by DNA vaccination, we hypothesized that a dual promoter plasmid encoding recombinant equine IL-12 (rEqIL-12) would function as a molecular adjuvant. In adult horses, DNA vaccines induced R. equi- and EIAV-specific antibody and lymphoproliferative responses, and EIAV-specific CTL and tetramer-positive CD8+ T lymphocytes. These responses were not enhanced by the rEqIL-12 plasmid. In neonatal foals, DNA immunization induced EIAV-specific antibody and lymphoproliferative responses, but not CTL. The R. equi vapA vaccine was poorly immunogenic in foals even when co-administered with the IL-12 plasmid. It was concluded that DNA immunization was capable of inducing Th1 responses in horses; dose and route were significant variables, but rEqIL-12 was not an effective molecular adjuvant. Additional work is needed to optimize DNA vaccine-induced Th1 responses in horses, especially in neonates.

  7. Prognosis and clinical evaluation of infection caused by Rhodococcus equi in HIV-infected patients: a multicenter study of 67 cases.

    PubMed

    Torres-Tortosa, Manuel; Arrizabalaga, Julio; Villanueva, José L; Gálvez, Juan; Leyes, María; Valencia, M Eulalia; Flores, Juan; Peña, José M; Pérez-Cecilia, Elisa; Quereda, Carmen

    2003-06-01

    To assess the clinical characteristics and the factors that influenced the prognosis of patients with HIV and infection caused by Rhodococcus equi. Observational, multicenter study in 29 Spanish general hospitals. These hospitals comprised a total of 20,250 beds for acute patients and served a population of 9,716,880 inhabitants. All patients with HIV and diagnosed R equi infection until September 1998. During the study period, 19,374 cases of AIDS were diagnosed. Sixty-seven patients were included (55 male patients; mean +/- SD age, 31.7 +/- 5.8 years). At the time of diagnosis of R equi infection, the mean CD4+ lymphocyte count was 35/ micro L (range, 1 to 183/ micro L) and the stage of HIV infection was A3 in 10.4% of patients, B3 in 31.3%, C3 in 56.7%, and unknown in 1.5%. R equi was most commonly isolated in sputum (52.2%), blood cultures (50.7%), and samples from bronchoscopy (31.3%). Chest radiographic findings were abnormal in 65 patients (97%). Infiltrates were observed in all of them, with cavitations in 45 patients. The most active antibiotics against the strains isolated were vancomycin, amikacin, rifampicin, imipenem, ciprofloxacin, and erythromycin. After a mean follow-up of 10.7 +/- 12.8 months, 23 patients (34.3%) died due to causes related to R equi infection and 6 other patients showed evidence of progression of the infection. The absence of highly active antiretroviral therapy (HAART) was independently associated with mortality related to R equi infection (relative risk, 53.4; 95% confidence interval, 1.7 to 1,699). Survival of patients treated with HAART was much higher than that of patients who did not receive this therapy. Infection by R equi is an infrequent, opportunistic complication of HIV infection and occurs during advanced stages of immunodepression. In these patients, it leads to a severe illness that usually causes a bacteremic, cavitary pneumonia, although HAART can improve the prognosis.

  8. The Pan-Genome of the Animal Pathogen Corynebacterium pseudotuberculosis Reveals Differences in Genome Plasticity between the Biovar ovis and equi Strains

    PubMed Central

    Soares, Siomar C.; Silva, Artur; Trost, Eva; Blom, Jochen; Ramos, Rommel; Carneiro, Adriana; Ali, Amjad; Santos, Anderson R.; Pinto, Anne C.; Diniz, Carlos; Barbosa, Eudes G. V.; Dorella, Fernanda A.; Aburjaile, Flávia; Rocha, Flávia S.; Nascimento, Karina K. F.; Guimarães, Luís C.; Almeida, Sintia; Hassan, Syed S.; Bakhtiar, Syeda M.; Pereira, Ulisses P.; Abreu, Vinicius A. C.; Schneider, Maria P. C.; Miyoshi, Anderson

    2013-01-01

    Corynebacterium pseudotuberculosis is a facultative intracellular pathogen and the causative agent of several infectious and contagious chronic diseases, including caseous lymphadenitis, ulcerative lymphangitis, mastitis, and edematous skin disease, in a broad spectrum of hosts. In addition, Corynebacterium pseudotuberculosis infections pose a rising worldwide economic problem in ruminants. The complete genome sequences of 15 C. pseudotuberculosis strains isolated from different hosts and countries were comparatively analyzed using a pan-genomic strategy. Phylogenomic, pan-genomic, core genomic, and singleton analyses revealed close relationships among pathogenic corynebacteria, the clonal-like behavior of C. pseudotuberculosis and slow increases in the sizes of pan-genomes. According to extrapolations based on the pan-genomes, core genomes and singletons, the C. pseudotuberculosis biovar ovis shows a more clonal-like behavior than the C. pseudotuberculosis biovar equi. Most of the variable genes of the biovar ovis strains were acquired in a block through horizontal gene transfer and are highly conserved, whereas the biovar equi strains contain great variability, both intra- and inter-biovar, in the 16 detected pathogenicity islands (PAIs). With respect to the gene content of the PAIs, the most interesting finding is the high similarity of the pilus genes in the biovar ovis strains compared with the great variability of these genes in the biovar equi strains. Concluding, the polymerization of complete pilus structures in biovar ovis could be responsible for a remarkable ability of these strains to spread throughout host tissues and penetrate cells to live intracellularly, in contrast with the biovar equi, which rarely attacks visceral organs. Intracellularly, the biovar ovis strains are expected to have less contact with other organisms than the biovar equi strains, thereby explaining the significant clonal-like behavior of the biovar ovis strains. PMID:23342011

  9. Rhodococcus equi pneumonia in foals: an assessment of the early diagnostic value of serum amyloid A and plasma fibrinogen concentrations in equine clinical practice.

    PubMed

    Passamonti, F; Vardi, D M; Stefanetti, V; Marenzoni, M L; Prato, S; Cévese, P; Coletti, M; Pepe, M; Casagrande Proietti, P; Olea-Popelka, F

    2015-02-01

    Early diagnosis and prevention of Rhodococcus equi pneumonia in foals represent important goals for equine clinicians. Recent protocols for diagnosis and treatment of Rhodococcosis in foals typically rely on a multimodal approach based on sonographic evidence suggestive of pyogranulomas, sonographic abscess scores and laboratory findings including plasma fibrinogen concentrations, blood biochemistry testing and platelet and leukocyte counts. The aim of this study was to assess the utility of weekly testing of serum amyloid A (SAA) and plasma fibrinogen concentrations in foals to achieve early diagnosis of R. equi pneumonia prior to the onset of clinical signs. This testing was used to simulate a clinically practical screening procedure and compared with thoracic ultrasonography performed in parallel. The present study suggests that SAA does not represent a reliable early marker of Rhodococcosis when plasma concentrations are tested weekly. However, when clinical signs of R. equi pneumonia are present, SAA concentrations may allow clinicians to obtain 'real-time' indications concerning both the progress of infection and the effectiveness of therapy. This study raises the possibility that plasma fibrinogen monitoring starting at 1 week of age and repeated on a weekly basis, could serve as a screening test allowing clinicians to identify foals as suspected of R. equi infection. Future investigations regarding both physiological plasma fibrinogen concentrations in foals as well as fibrinogen kinetics in foals affected with R. equi pneumonia, including the establishment of appropriate reference intervals for the test method employed in this study, will be necessary in order to clarify this possibility. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Vaccination of Mice with Salmonella Expressing VapA: Mucosal and Systemic Th1 Responses Provide Protection against Rhodococcus equi Infection

    PubMed Central

    Oliveira, Aline F.; Ruas, Luciana P.; Cardoso, Silvia A.; Soares, Sandro G.; Roque-Barreira, Maria-Cristina

    2010-01-01

    Conventional vaccines to prevent the pneumonia caused by Rhodococcus equi have not been successful. We have recently demonstrated that immunization with Salmonella enterica Typhimurium expressing the VapA antigen protects mice against R. equi infection. We now report that oral vaccination of mice with this recombinant strain results in high and persistent fecal levels of antigen-specific IgA, and specific proliferation of the spleen cells of immunized mice in response to the in vitro stimulation with R. equi antigen. After in vitro stimulation, spleen cells of immunized mice produce high levels of Th1 cytokines and show a prominent mRNA expression of the Th1 transcription factor T-bet, in detriment of the Th2 transcription factor GATA-3. Following R. equi challenge, a high H2O2, NO, IL-12, and IFN-γ content is detected in the organs of immunized mice. On the other hand, TNF-α and IL-4 levels are markedly lower in the organs of vaccinated mice, compared with the non-vaccinated ones. The IL-10 content and the mRNA transcription level of TGF-β are also higher in the organs of immunized mice. A greater incidence of CD4+ and CD8+ T cells and B lymphocytes is verified in vaccinated mice. However, there is no difference between vaccinated and non-vaccinated mice in terms of the frequency of CD4+CD25+Foxp3+ T cells. Finally, we show that the vaccination confers a long-term protection against R. equi infection. Altogether, these data indicate that the oral vaccination of mice with S. enterica Typhimurium expressing VapA induces specific and long-lasting humoral and cellular responses against the pathogen, which are appropriately regulated and allow tissue integrity after challenge. PMID:20072623

  11. A case-control study developing a model for predicting risk factors for high SeM-specific antibody titers after natural outbreaks of Streptococcus equi subsp equi infection in horses.

    PubMed

    Boyle, Ashley G; Smith, Meagan A; Boston, Raymond C; Stefanovski, Darko

    2017-06-15

    OBJECTIVE To develop a risk prediction model for factors associated with an SeM-specific antibody titer ≥ 3,200 in horses after naturally occurring outbreaks of Streptococcus equi subsp equi infection and to validate this model. DESIGN Case-control study. ANIMALS 245 horses: 57 horses involved in strangles outbreaks (case horses) and 188 healthy horses (control horses). PROCEDURES Serum samples were obtained from the 57 cases over a 27.5-month period after the start of outbreaks; serum samples were obtained once from the 188 controls. A Bayesian mixed-effects logistic regression model was used to assess potential risk factors associated with an antibody titer ≥ 3,200 in the case horses. A cutoff probability for an SeM-specific titer ≥ 3,200 was determined, and the model was externally validated in the control horses. Only variables with a 95% credibility interval that did not overlap with a value of 1 were considered significant. RESULTS 9 of 57 (6%) case horses had at least 1 titer ≥ 3,200, and 7 of 188 (3.7%) of control horses had a titer ≥ 3,200. The following variables were found to be significantly associated with a titer ≥ 3,200 in cases: farm size > 20 horses (OR, 0.11), history of clinically evident disease (OR, 7.92), and male sex (OR, 0.11). The model had 100% sensitivity but only 24% specificity when applied to the 188 control horses (area under the receiver operating characteristic curve = 0.62.) CONCLUSIONS AND CLINICAL RELEVANCE Although the Bayesian mixed-effects logistic regression model developed in this study did not perform well, it may prove useful as an initial screening tool prior to vaccination. We suggest that SeM-specific antibody titer be measured prior to vaccination when our model predicts a titer ≥ 3,200.

  12. Identification of Virulence-Associated Plasmids in Rhodococcus equi in Humans with and without Acquired Immunodeficiency Syndrome in Brazil

    PubMed Central

    Ribeiro, Márcio Garcia; Takai, Shinji; de Vargas, Agueda Castagna; Mattos-Guaraldi, Ana Luiza; Ferreira Camello, Thereza Cristina; Ohno, Ryoko; Okano, Hajime; da Silva, Aristeu Vieira

    2011-01-01

    Virulence of Rhodococcus equi strains from 20 humans in Brazil was investigated by using a polymerase chain reaction to characterize isolates as virulent (VapA), intermediately virulent (VapB), and avirulent. Nine isolates were obtained from human immunodeficiency virus (HIV)–positive patients, six from HIV-negative patients, and five from patients of unknown status. Five isolates were VapB positive, four were VapA positive, and eleven were avirulent. Among the nine isolates from HIV-positive patients, five contained VapB plasmids and two contained VapA plasmids. Five VapB-positive isolates had the type 8 virulence plasmid. Eleven of the patients had a history of contact with livestock and/or a farm environment, and none had contact with pigs. PMID:21896813

  13. Presumptive meningoencephalitis secondary to extension of otitis media/interna caused by Streptococcus equi subspecies zooepidemicus in a cat.

    PubMed

    Martin-Vaquero, Paula; da Costa, Ronaldo C; Daniels, Joshua B

    2011-08-01

    A 5-year-old castrated male domestic longhair cat was presented with neurological signs consistent with a central vestibular lesion and left Horner's syndrome. Computed tomography images revealed hyperattenuating, moderately contrast-enhancing material within the left tympanic bulla, most consistent with left otitis media/interna. Marked neutrophilic pleocytosis was identified on cerebrospinal fluid analysis. Streptococcus equi subspecies zooepidemicus (SEZ) was isolated from the cerebrospinal fluid. Intracranial extension of otitis media/interna is relatively infrequent in small animals. There are no reports of otitis media/interna caused by SEZ in dogs or cats. This is the first report of otitis media/interna and presumptive secondary meningoencephalitis caused by SEZ in a cat. Copyright © 2011 ISFM and AAFP. Published by Elsevier Ltd. All rights reserved.

  14. The horse pinworm (Oxyuris equi) in archaeology during the Holocene: Review of past records and new data.

    PubMed

    Dufour, Benjamin; Hugot, Jean-Pierre; Lepetz, Sébastien; Le Bailly, Matthieu

    2015-07-01

    This paper focuses on the horse pinworm, Oxyuris equi, in archaeology during the Holocene period, and presents an overview of past published occurrences, early mentions in texts, and new data from our paleoparasitology research. This original compilation shows that the most ancient record of the horse pinworm dates to ca. 2500 years before present (ybp) in Central Asia and to ca. 2020 ybp in Western Europe. It also shows that the parasite is not detected on the American continent until contemporary periods. The role of European migrations from 1492 (Christopher Columbus) is discussed to explain the transfer of the horse pinworm from the Old World to the Americas. The absence of any record of this parasite before ca. 2500 ybp in Eurasia could be explained by parasite ecology, unfavorable sampling and scarcity of horse archeological remains. For the Americas, the absence of horse for long periods can be an additional explanation for the absence of the parasite.

  15. Elucidation of simple pathways for reconstructive phase transitions using periodic equi-surfaces (PES) descriptors. II. The strontium disilicide transition

    NASA Astrophysics Data System (ADS)

    Leoni, Stefano; Nesper, Reinhard

    2003-01-01

    A short topological pathway relating two fundamental three-dimensional, three-connected networks as they are found in α-ThSi 2 and SrSi 2 is proposed. The approach exploits a Fourier function approach based on periodic nodal (PNS) and periodic equi-surface (PES) descriptors. Continuous functions are chosen to represent the networks in question, and a linear transition approach provides a transition model. On this basis, each network is fragmented into a collection of dumb-bells, and the transition is described in terms of a concerted reorientation of the latter. The centers of mass of the dumb-bells are shown to conserve the diamond topology at each stage of the transition, as an underlying motif of the whole process.

  16. Effect of nitrous oxide on bispectral index values at equi-minimum alveolar concentrations of sevoflurane and desflurane.

    PubMed

    Mishra, Rajeeb Kumar; Mahajan, Charu; Prabhakar, Hemanshu; Kapoor, Indu; Bithal, Parmod Kumar

    2017-06-01

    Bispectral index (BIS) values may be anaesthetic agent-specific, depending on their ability to suppress the electroencephalogram (EEG) signals. We carried out a prospective, randomised clinical trial to study the effect of nitrous oxide (N2O) on the BIS values at an equi-minimum alveolar concentration (MAC) of sevoflurane and desflurane. Sixty adult patients undergoing spine surgery were randomised into two groups; Group S (sevoflurane; n = 30) and Group D (desflurane; n = 30) for the maintenance of anaesthesia in oxygen and air or oxygen and N2O mixture (FiO2-0.4) (Stage 1). BIS and fraction of inspired and end-tidal concentration of agents were noted at 1.0 MAC. In Stage 2, air or N2O was discontinued and the other carrier gas was introduced. At steady state of this carrier gas, values were again noted as in Stage 1. Statistical analysis was performed using two-way analysis of variance followed by Bonferroni correction, and Student's t-test for paired data. P<0.05 was considered statistically significant. With air-oxygen as the carrier gas, sevoflurane and desflurane resulted in comparable BIS values (P = 0.44). With addition of 60% N2O, there was a significant increase in BIS values at 1.0 MAC for both the agents. Furthermore, higher BIS values were observed with sevoflurane compared to desflurane (P = 0.01). Sevoflurane and desflurane at equi-MAC concentration exert similar effect on BIS values when used with air-oxygen. N2O results in higher BIS values; this effect is more pronounced in combination with sevoflurane.

  17. Standardisation and comparison of serial dilution and single dilution enzyme linked immunosorbent assay (ELISA) using different antigenic preparations of the Babesia (Theileria) equi parasite.

    PubMed

    Kumar, Sanjay; Kumar, Yogesh; Malhotra, Dharam V; Dhar, Shruti; Nichani, Anil K

    2003-01-01

    Serial dilution and single dilution enzyme linked immunosorbent assays (ELISA) were standardised and their sensitivity and specificity were compared for serodiagnosis of Babesia equi infection. The antibody titres of 24 donkey sera of known identity were determined separately by serial dilution ELISA using three different B. equi antigens namely whole merozoite (WM), cell membrane (CM) and high speed supernatant (HSS). The ratios of the optical density (OD) of known positive and known negative sera at different serum dilutions were calculated and termed as the positive/negative (P/N) ratio. The coefficients of correlation (r) were calculated between the P/N ratios at different dilutions of sera and the log10 antibody titres of the same sera were ascertained by serial dilution ELISA. The highest value of 'r' was obtained at a serum dilution of 1:200. From log10 antibody titre of sera (y) and their P/N ratio at a dilution of 1:200 (x), regression equations (y = a + bx) were calculated separately for the three antigens. Test sera were diluted to 1:200, their OD were read in duplicate wells and were converted to the P/N ratio. Antibody titres were predicted from the P/N ratio using a regression equation separately for the three antigens. Titres obtained by both ELISAs were not significantly different from each other, thus confirming that single dilution ELISA could be successfully used to replace conventional serial dilution ELISA. The sensitivity, specificity and predictive value of single dilution ELISA was validated statistically using 42 B. equi disease-positive sera and 106 B. equi disease-negative sera. The WM antigen was found to be the most sensitive with a higher predictive value for negative test sera as compared to the CM or HSS antigens. Sera positive for other equine infections including Babesia caballi showed no cross-reaction with the three B. equi antigens in ELISA, thus the test was immunologically specific. Antibody titres of 109 unknown field donkey

  18. Rhodococcus defluvii sp. nov., isolated from wastewater of a bioreactor and formal proposal to reclassify [Corynebacterium hoagii] and Rhodococcus equi as Rhodococcus hoagii comb. nov.

    PubMed

    Kämpfer, P; Dott, W; Martin, K; Glaeser, S P

    2014-03-01

    A Gram-stain-positive, non-endospore-forming rod-shaped bacterium, strain Ca11(T), was isolated from a bioreactor with extensive phosphorus removal and was studied in detail for its taxonomic allocation. 16S rRNA gene sequence analysis revealed closest sequence similarity of the strain to type strains of [Corynebacterium hoagii] and Rhodococcus equi (98.9%), Rhodococcus koreensis and Rhodococcus wratislaviensis (both 98.4%), Rhodococcus opacus and Rhodococcus canchipurensis (both 98.0%) followed by Rhodococcus kunmingensis and Rhodococcus imtechensis (97.7%). Phylogenetic trees showed a distinct clustering of strain Ca11(T) with the type strains of [C. hoagii], R. equi, and R. kunmingensis separate to all other species of the genus Rhodococcus. The quinone system of strain Ca11(T) was composed of dihydrogenated menaquinones with 8 (major amount) as well as 7 and 6 isoprenoid units [MK-8(H2), MK-7(H2), MK-6(H2)]. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside, one unknown phospholipid and an unidentified glycolipid. The fatty acid profile was similar to that reported for R. equi and contained major amounts of C16:0, C18:1ω9c and 10-methyl C18:0, supporting the allocation of the strain to the genus Rhodococcus. Physiological and biochemical characterization and DNA-DNA hybridization with type strains of the most closely related species allowed clear phenotypic and genotypic differentiation of the isolate. On the basis of these results, strain Ca11(T) ( = DSM 45893(T) = LMG 27563(T)) represents a novel species of the genus Rhodococcus, with the proposed name Rhodococcus defluvii sp. nov. In addition, a polyphasic taxonomic analysis of [Corynebacterium hoagii] DSM 20295(T) and Rhodococcus equi DSM 20307(T) indicated that the two strains belong to the same species, for which the name Rhodococcus hoagii comb. nov. takes priority, according to the Rules of the

  19. Analysis of plasmid diversity in 96 Rhodococcus equi strains isolated in Normandy (France) and sequencing of the 87-kb type I virulence plasmid.

    PubMed

    Duquesne, Fabien; Hébert, Laurent; Sévin, Corinne; Breuil, Marie-France; Tapprest, Jackie; Laugier, Claire; Petry, Sandrine

    2010-10-01

    To characterize the potential epidemiological relationship between the origin of Rhodococcus equi strains and the type of their virulence plasmids, we performed a comparative analysis of virulence plasmid types encountered in 96 R. equi strains isolated from (1) autopsied horses, (2) organic samples (horse faeces, manure and straw) and (3) environmental samples. Our results revealed no clear epidemiological link between virulence plasmid type and the origin of R. equi strains isolated from horse-related environments. To understand this result, we determined the nucleotide sequence of the second most frequently isolated virulence plasmid type: a 87-kb type I (pVAPA116) plasmid and compared it with the previously sequenced (and most commonly encountered) 85-kb type I (pVAPA1037) plasmid. Our results show that the divergence between these two plasmids is mainly due to the presence of three allelic exchange loci, resulting in the deletion of two genes and the insertion of three genes in pVAPA116 compared with pVAPA1037. In conclusion, it appears that the divergence between the two sequenced rhodococcal virulence plasmids is not associated with the vap pathogenicity island and may result from an evolutionary process driven by a mobility-related invertase/resolvase invA-like gene. © 2010 ANSES. Journal compilation © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd.

  20. VirS, an OmpR/PhoB subfamily response regulator, is required for activation of vapA gene expression in Rhodococcus equi.

    PubMed

    Kakuda, Tsutomu; Hirota, Takuya; Takeuchi, Tatsuya; Hagiuda, Hirofumi; Miyazaki, Shiko; Takai, Shinji

    2014-10-03

    Rhodococcus equi is an important pulmonary pathogen in foals and in immunocompromised individuals. Virulent R. equi strains carry an 80-90 kb virulence plasmid that expresses the virulence-associated protein A (VapA). VapA expression is regulated by temperature and pH. The LysR-type transcriptional regulator, VirR, is involved in the regulation of the vapA gene. To examine the mechanism underlying transcriptional regulation of vapA, we characterized an R. equi mutant in which another putative transcriptional regulator encoded on the virulence plasmid, VirS, was deleted. Deletion of virS reduced vapA promoter activity to non-inducible levels. Complementary expression of VirS in the virS deletion mutant restored transcription at the PvapA promoter, even under non-inducing conditions (30°C and pH 8.0). In addition, VirS expression increased PvapA promoter activity in the absence of functional VirR. Further, transcription of the icgA operon containing virS was regulated by pH and temperature in the same manner as vapA. This study suggests that VirS is required for VapA expression and that regulation of PvapA-promoter activity may be achieved by controlling VirS expression levels.

  1. Targeted thrombolysis of tissue plasminogen activator and streptokinase with extracellular biosynthesis nanoparticles using optimized Streptococcus equi supernatant.

    PubMed

    Tadayon, Ateke; Jamshidi, Reza; Esmaeili, Akbar

    2016-03-30

    Extracellular biosynthesis of nanoparticles have many important advantages such as well dispersed in aqueous solutions, low energy requirements, ecofriendly, non-toxic, low-costs and non-flocculate. This technique have shown significant promise as targeted drug delivery applications. In this investigation, for the first time, we examine the efficacy of targeted therapeutic delivery with t-PA and SK immobilized to biosynthesis of nanoparticles (CuNP) by using Streptococcus equi strains isolated from the horses of Iran and their ability to produce metallic nanoparticles. Also we compared them with their chemical synthesis. The S. equi was screened for its ability to produce MNPs. The minimum size and shapes (23-89 nm) are presented in the formation with good dispersion and high stability. Response Surface methodology was applied for the optimized production of biological CuNPs. The growth factors like pH, temperature and incubation time was changed. The optimum conditions to obtain CuNPs were found with the culture conditions of pH 7.5 in 120 h at 35 °C. To determine some of MNPs structural properties UV-vis absorption spectrophotometer, FTIR, XRD and SEM has characterized. The results provided some parameters may impact on the formation of biological MNPs. Lastly, these MNPs were conjugated with t-PA and SK, as a drug carrier. In addition, effective thrombolysis with magnet-guided SiO2CuNPs-tPA-SK is demonstrated in rat embolism model where 18.6% of the regular t-PA dose and 15.78% of SK dose restored and 15-25 min reductions in blood clot lysis time were observed compared with runs with free t-PA and without magnet-guided and using the same drug dosage. The comparison between CuNPs with MNPs shows that thrombolysis had not been directed to the type of magnetic carrier under the magnetic guide. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. The arginine deiminase system facilitates environmental adaptability of Streptococcus equi ssp. zooepidemicus through pH adjustment.

    PubMed

    Xu, Bin; Yang, Xinyi; Zhang, Ping; Ma, Zhe; Lin, Huixing; Fan, Hongjie

    2016-06-01

    The arginine deiminase system (ADS) is a secondary metabolic system found in many different bacterial pathogens and it is often associated with virulence. Here, a systematic study of ADS functions in Streptococcus equi subsp. zooepidemicus (SEZ) was performed. Transcriptional levels of ADS operon genes were observed to be significantly increased when SEZ was grown under acidic conditions. We constructed arcA and arcD deletion mutants (SEZ ΔarcA and SEZ ΔarcD, respectively) and found that SEZ ΔarcA was unable to metabolize arginine and synthesize ammonia; however, arcD deletion resulted in an initial decrease in arginine consumption and ammonia production, followed by recovery to the levels of wild-type SEZ after 24 h of cultivation. Cell extracts of SEZ ΔarcA showed no arginine deiminase (AD) activity, whereas no difference in AD activity between SEZ ΔarcD and wild-type SEZ was observed. SEZ survival tests demonstrated a significant decrease in survival for SEZ ΔarcA, when compared with wild-type SEZ, under acidic conditions and in epithelial cells. These findings indicate that ADS in SEZ contributes to environmental adaptability via ammonia synthesis to reduce pH stress.

  3. Engineering S. equi subsp. zooepidemicus towards concurrent production of hyaluronic acid and chondroitin biopolymers of biomedical interest.

    PubMed

    Cimini, Donatella; Iacono, Ileana Dello; Carlino, Elisabetta; Finamore, Rosario; Restaino, Odile F; Diana, Paola; Bedini, Emiliano; Schiraldi, Chiara

    2017-12-01

    Glycosaminoglycans, such as hyaluronic acid and chondroitin sulphate, are not only more and more required as main ingredients in cosmeceutical and nutraceutical preparations, but also as active principles in medical devices and pharmaceutical products. However, while biotechnological production of hyaluronic acid is industrially established through fermentation of Streptococcus spp. and recently Bacillus subtilis, biotechnological chondroitin is not yet on the market. A non-hemolytic and hyaluronidase negative S. equi subsp. zooepidemicus mutant strain was engineered in this work by the addition of two E. coli K4 genes, namely kfoA and kfoC, involved in the biosynthesis of chondroitin-like polysaccharide. Chondroitin is the precursor of chondroitin sulphate, a nutraceutical present on the market as anti-arthritic drug, that is lately being studied for its intrinsic bioactivity. In small scale bioreactor batch experiments the production of about 1.46 ± 0.38 g/L hyaluronic acid and 300 ± 28 mg/L of chondroitin with an average molecular weight of 1750 and 25 kDa, respectively, was demonstrated, providing an approach to the concurrent production of both biopolymers in a single fermentation.

  4. Efficacy of a 2-dose regimen of a sustained release ceftiofur suspension in horses with Streptococcus equi subsp. zooepidemicus bronchopneumonia.

    PubMed

    McClure, S; Sibert, G; Hallberg, J; Bade, D

    2011-10-01

    The efficacy and safety of sustained release ceftiofur administered twice, 4 days apart, for treatment of horses with naturally acquired Streptococcus equi subsp. zooepidemicus (Strep. zoo.) pneumonia was evaluated in a multicenter, placebo-controlled, double-blinded, randomized clinical trial. The study included 373 horses (278 treated and 95 placebos) with naturally acquired pneumonia. Inclusion in the statistical analyses for treatment efficacy for Strep. zoo. required recovery of ≥10(4) CFU/mL of Strep. zoo. on the primary isolation plate which resulted in 201 cases (145 treated and 56 placebos) with confirmed Strep. zoo. pneumonia evaluable for treatment success. Therapeutic success was defined by clinical improvement of lower respiratory tract infection at 4 and 9 days after initial dosing, resolution of clinical signs by 15 days, and no recurrence by 25 days. Of the 278 treated horses, 239 (85.9%) completed the 25 day study without additional therapy compared to 50 of the 95 (53.6%) placebo horses. In confirmed Strep. zoo. cases, a clinical cure was achieved in 66.9% of 145 treated horses compared to 32.1% of 56 placebo horses (P = 0.0286). Two doses of sustained release ceftiofur suspension were effective and safe in the treatment of naturally acquired lower respiratory tract infection associated with Strep. zoo. in horses under field use conditions.

  5. Virulent and Vaccine Strains of Streptococcus equi ssp. zooepidemicus Have Different Influences on Phagocytosis and Cytokine Secretion of Macrophages.

    PubMed

    Jie, Peng; Zhe, Ma; Chengwei, Hua; Huixing, Lin; Hui, Zhang; Chengping, Lu; Hongjie, Fan

    2017-01-06

    Swine streptococcosis is a significant threat to the Chinese pig industry, and Streptococcus equi ssp. zooepidemicus (SEZ) is one of the major pathogens. SEZ ATCC35246 is a classical virulent strain, while SEZ ST171 is a Chinese attenuated vaccine strain. In this study, we employed stable isotope labeling by amino acids in cell culture and liquid chromatography-mass spectrometry (LC-MS) to determine the differential response of macrophages to infection by these two strains. Eighty-seven upregulated proteins and 135 downregulated proteins were identified. The proteomic results were verified by real-time polymerase chain reaction for 10 chosen genes and Western blotting for three proteins. All differentially abundant proteins were analyzed for their Gene Ontology and Kyoto Encyclopedia of Genes and Genomes annotations. Certain downregulated proteins were associated with immunity functions, and the upregulated proteins were related to cytomembrane and cytoskeleton regulation. The phagocytosis rate and cytokine genes transcription in Raw264.7 cells during SEZ ATCC35246 and ST171 infection were detected to confirm the bioinformatics results. These results showed that different effects on macrophage phagocytosis and cytokine expression might explain the different phenotypes of SEZ ATCC35246 and ST171 infection. This research provided clues to the mechanisms of host immunity responses to SEZ ST171and SEZ ATCC35246, which could identify potential therapy and vaccine development targets.

  6. A novel suicide shuttle plasmid for Streptococcus suis serotype 2 and Streptococcus equi ssp. zooepidemicus gene mutation

    PubMed Central

    Liu, Rui; Zhang, Ping; Su, Yiqi; Lin, Huixing; Zhang, Hui; Yu, Lei; Ma, Zhe; Fan, Hongjie

    2016-01-01

    The mariner-based Himar1 system has been utilized for creating mutant libraries of many Gram-positive bacteria. Streptococcus suis serotype 2 (SS2) and Streptococcus equi ssp. zooepidemicus (SEZ) are primary pathogens of swine that threaten the swine industry in China. To provide a forward-genetics technology for finding virulent phenotype-related genes in these two pathogens, we constructed a novel temperature-sensitive suicide shuttle plasmid, pMar4s, which contains the Himar1 system transposon, TnYLB-1, and the Himar1 C9 transposase from pMarA and the repTAs temperature-sensitive fragment from pSET4s. The kanamycin (Kan) resistance gene was in the TnYLB-1 transposon. Temperature sensitivity and Kan resistance allowed the selection of mutant strains and construction of the mutant library. The SS2 and SEZ mutant libraries were successfully constructed using the pMar4s plasmid. Inverse-Polymerase Chain Reaction (Inverse-PCR) results revealed large variability in transposon insertion sites and that the library could be used for phenotype alteration screening. The thiamine biosynthesis gene apbE was screened for its influence on SS2 anti-phagocytosis; likewise, the sagF gene was identified to be a hemolytic activity-related gene in SEZ. pMar4s was suitable for mutant library construction, providing more information regarding SS2 and SEZ virulence factors and illustrating the pathogenesis of swine streptococcosis. PMID:27256117

  7. The Equine Antimicrobial Peptide eCATH1 Is Effective against the Facultative Intracellular Pathogen Rhodococcus equi in Mice

    PubMed Central

    Schlusselhuber, Margot; Torelli, Riccardo; Martini, Cecilia; Leippe, Matthias; Cattoir, Vincent; Leclercq, Roland; Laugier, Claire; Grötzinger, Joachim; Sanguinetti, Maurizio

    2013-01-01

    Rhodococcus equi, the causal agent of rhodococcosis, is a major pathogen of foals and is also responsible for severe infections in immunocompromised humans. Of great concern, strains resistant to currently used antibiotics have emerged. As the number of drugs that are efficient in vivo is limited because of the intracellular localization of the bacterium inside macrophages, new active but cell-permeant drugs will be needed in the near future. In the present study, we evaluated, by in vitro and ex vivo experiments, the ability of the alpha-helical equine antimicrobial peptide eCATH1 to kill intracellular bacterial cells. Moreover, the therapeutic potential of the peptide was assessed in experimental rhodococcosis induced in mice, while the in vivo toxicity was evaluated by behavioral and histopathological analysis. The study revealed that eCATH1 significantly reduced the number of bacteria inside macrophages. Furthermore, the bactericidal potential of the peptide was maintained in vivo at doses that appeared to have no visible deleterious effects for the mice even after 7 days of treatment. Indeed, daily subcutaneous injections of 1 mg/kg body weight of eCATH1 led to a significant reduction of the bacterial load in organs comparable to that obtained after treatment with 10 mg/kg body weight of rifampin. Interestingly, the combination of the peptide with rifampin showed a synergistic interaction in both ex vivo and in vivo experiments. These results emphasize the therapeutic potential that eCATH1 represents in the treatment of rhodococcosis. PMID:23817377

  8. The impact of membrane lipid composition on macrophage activation in the immune defense against Rhodococcus equi and Pseudomonas aeruginosa.

    PubMed

    Schoeniger, Axel; Adolph, Stephanie; Fuhrmann, Herbert; Schumann, Julia

    2011-01-01

    Nutritional fatty acids are known to have an impact on membrane lipid composition of body cells, including cells of the immune system, thus providing a link between dietary fatty acid uptake, inflammation and immunity. In this study we reveal the significance of macrophage membrane lipid composition on gene expression and cytokine synthesis thereby highlighting signal transduction processes, macrophage activation as well as macrophage defense mechanisms. Using RAW264.7 macrophages as a model system, we identified polyunsaturated fatty acids (PUFA) of both the n-3 and the n-6 family to down-regulate the synthesis of: (i) the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α; (ii) the co-stimulatory molecule CD86; as well as (iii) the antimicrobial polypeptide lysozyme. The action of the fatty acids partially depended on the activation status of the macrophages. It is particularly important to note that the anti-inflammatory action of the PUFA could also be seen in case of infection of RAW264.7 with viable microorganisms of the genera R. equi and P. aeruginosa. In summary, our data provide strong evidence that PUFA from both the n-3 and the n-6 family down-regulate inflammation processes in context of chronic infections caused by persistent pathogens.

  9. Genomic and Functional Analyses of Rhodococcus equi Phages ReqiPepy6, ReqiPoco6, ReqiPine5, and ReqiDocB7 ▿

    PubMed Central

    Summer, E. J.; Liu, M.; Gill, J. J.; Grant, M.; Chan-Cortes, T. N.; Ferguson, L.; Janes, C.; Lange, K.; Bertoli, M.; Moore, C.; Orchard, R. C.; Cohen, N. D.; Young, R.

    2011-01-01

    The isolation and results of genomic and functional analyses of Rhodococcus equi phages ReqiPepy6, ReqiDocB7, ReqiPine5, and ReqiPoco6 (hereafter referred to as Pepy6, DocB7, Pine5, and Poco6, respectively) are reported. Two phages, Pepy6 and Poco6, more than 75% identical, exhibited genome organization and protein sequence likeness to Lactococcus lactis phage 1706 and clostridial prophage elements. An unusually high fraction, 27%, of Pepy6 and Poco6 proteins were predicted to possess at least one transmembrane domain, a value much higher than the average of 8.5% transmembrane domain-containing proteins determined from a data set of 36,324 phage protein entries. Genome organization and protein sequence comparisons place phage Pine5 as the first nonmycobacteriophage member of the large Rosebush cluster. DocB7, which had the broadest host range among the four isolates, was not closely related to any phage or prophage in the database, and only 23 of 105 predicted encoded proteins could be assigned a functional annotation. Because of the relationship of Rhodococcus to Mycobacterium, it was anticipated that these phages should exhibit some of the features characteristic of mycobacteriophages. Traits that were identified as shared by the Rhodococcus phages and mycobacteriophages include the prevalent long-tailed morphology and the presence of genes encoding LysB-like mycolate-hydrolyzing lysis proteins. Application of DocB7 lysates to soils amended with a host strain of R. equi reduced recoverable bacterial CFU, suggesting that phage may be useful in limiting R. equi load in the environment while foals are susceptible to infection. PMID:21097585

  10. VapA-specific IgG and IgG subclasses responses after natural infection and experimental challenge of foals with Rhodococcus equi.

    PubMed

    Sanz, M G; Villarino, N; Ferreira-Oliveira, A; Horohov, D W

    2015-03-15

    Rhodococcus equi is a common cause of pneumonia in young foals worldwide and has considerable economic effects on the global equine industry. Despite ongoing efforts, no vaccine is currently available to prevent rhodococaal pneumonia. This is due, in part, to an incomplete understanding of the protective immune response to this bacterium. While antibodies to VapA, a lipoprotein produced by virulent R. equi, are useful in differentiating antibody production in response to pathogenic versus non-pathogenic strains, the significance of the humoral response of foals to this lipoprotein remains poorly defined. The objectives of this study were to evaluate changes in VapA-specific IgG and IgG subclasses after exposure and infection of neonatal foals. Experimental foals included those challenged with R. equi at 1 (n=18), 2 (n=4) and 3 (n=6) weeks of age. Confirmed naturally infected (n=7) and not infected (n=3) foals were also included. All foals were bled 24h after birth and weekly thereafter for a period of 8 weeks. Antibody changes over time were evaluated. Following birth, VapA-specific IgGs significantly (p<0.05) decreased over time in all foals as a result of normal decay of passively transferred antibodies. Both VapA-specific IgGa and IgG(T) significantly increased (p<0.05) after experimental challenge, however, the rise in IgG(T) occurred earlier. Only a significant (p<0.05) increase in VapA-specific IgG(T) over time was seen after natural infection. Whether VapA-specific IgG(T) can be used to differentiate rhodococcal from other pneumonias requires further investigation under field conditions. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Evolution of the Rhodococcus equi vap pathogenicity island seen through comparison of host-associated vapA and vapB virulence plasmids.

    PubMed

    Letek, Michal; Ocampo-Sosa, Alain A; Sanders, Mandy; Fogarty, Ursula; Buckley, Tom; Leadon, Desmond P; González, Patricia; Scortti, Mariela; Meijer, Wim G; Parkhill, Julian; Bentley, Stephen; Vázquez-Boland, José A

    2008-09-01

    The pathogenic actinomycete Rhodococcus equi harbors different types of virulence plasmids associated with specific nonhuman hosts. We determined the complete DNA sequence of a vapB(+) plasmid, typically associated with pig isolates, and compared it with that of the horse-specific vapA(+) plasmid type. pVAPB1593, a circular 79,251-bp element, had the same housekeeping backbone as the vapA(+) plasmid but differed over an approximately 22-kb region. This variable region encompassed the vap pathogenicity island (PAI), was clearly subject to selective pressures different from those affecting the backbone, and showed major genetic rearrangements involving the vap genes. The pVAPB1593 PAI harbored five different vap genes (vapB and vapJ to -M, with vapK present in two copies), which encoded products differing by 24 to 84% in amino acid sequence from the six full-length vapA(+) plasmid-encoded Vap proteins, consistent with a role for the specific vap gene complement in R. equi host tropism. Sequence analyses, including interpolated variable-order motifs for detection of alien DNA and reconstruction of Vap family phylogenetic relationships, suggested that the vap PAI was acquired by an ancestor plasmid via lateral gene transfer, subsequently evolving by vap gene duplication and sequence diversification to give different (host-adapted) plasmids. The R. equi virulence plasmids belong to a new family of actinobacterial circular replicons characterized by an ancient conjugative backbone and a horizontally acquired niche-adaptive plasticity region.

  12. Selective pressure for allelic diversity in SeM of Streptococcus equi does not affect immunoreactive proteins SzPSe or Se18.9.

    PubMed

    Ijaz, Muhammad; Velineni, Sridhar; Timoney, John F

    2011-07-01

    Streptococcus equi, a clone or biovar of an ancestral Streptococcus zooepidemicus of Lancefield group C causes equine strangles, a highly contagious tonsillitis and lymphadenitis of the head and neck. At least 74 alleles based on N-terminal amino acid sequence of the anti-phagocytic SeM have been observed among isolates of S. equi from N. America, Europe and Japan. A d(N)/d(S) ratio of 5.93 for the 5' region of sem is indicative of positive selective pressure. The aim of this study was to determine whether variations in SeM were accompanied by variations in the surface exposed SzPSe and secreted Se18.9, both of which bind to equine tonsillar epithelium and, along with SeM, elicit strong nasopharyngeal IgA responses during convalescence. Sequences of genes for these proteins from 25 S. equi expressing 19 different SeM alleles isolated over 40 years in different countries were compared. No variation was observed in szpse, except for an Australian isolate with a deletion of a single repeat in the 3' end of the gene. Interestingly, only two SNP loci were detected in se18.9 compared to 93 and 55 in sem and szpse, respectively. The high frequency of nucleotide substitutions in szpse may be related to its mosaic structure since this gene in S. zooepidemicus exists in a variety of combinations of sequence segments and has a central hypervariable region that includes exogenous DNA sequence based on an atypical G-C percentage. In summary, the results of this study document very different responses of streptococcal genes for 3 immunoreactive proteins to selection pressure of the nasopharyngeal mucosal immune response. Copyright © 2011 Elsevier B.V. All rights reserved.

  13. First record of protozoan parasites, Tetrahymena rostrata and Callimastix equi from the edible oyster in Sundarbans region of West Bengal, India.

    PubMed

    Biswas, Tanima; Bandyopadhyay, Probir Kumar

    2016-09-01

    Several protozoan parasites have been found infecting the edible oysters, hence deteriorating the meat quality. Protozoan parasites such as, Tetrahymena rostrata and Callimastix equi infested the edible oyster in Sundarbans region, West Bengal, India, are first record from this region. Due to filter feeding habit of the organisms, oysters provides excellent ecological services in regard to efficient cleaning of infectious agents from surrounding water as a potential measure to improve water quality. However, these environmental benefits are associated with public heath risks from contaminated oysters intended for human consumption.

  14. Structural characterisation of the virulence-associated protein VapG from the horse pathogen Rhodococcus equi

    PubMed Central

    Okoko, Tebekeme; Blagova, Elena V.; Whittingham, Jean L.; Dover, Lynn G.; Wilkinson, Anthony J.

    2015-01-01

    Virulence and host range in Rhodococcus equi depends on the variable pathogenicity island of their virulence plasmids. Notable gene products are a family of small secreted virulence-associated proteins (Vaps) that are critical to intramacrophagic proliferation. Equine-adapted strains, which cause severe pyogranulomatous pneumonia in foals, produce a cell-associated VapA that is necessary for virulence, alongside five other secreted homologues. In the absence of biochemical insight, attention has turned to the structures of these proteins to develop a functional hypothesis. Recent studies have described crystal structures for VapD and a truncate of the VapA orthologue of porcine-adapted strains, VapB. Here, we crystallised the full-length VapG and determined its structure by molecular replacement. Electron density corresponding to the N-terminal domain was not visible suggesting that it is disordered. The protein core adopted a compact elliptical, anti-parallel β-barrel fold with β1–β2–β3–β8–β5–β6–β7–β4 topology decorated by a single peripheral α-helix unique to this family. The high glycine content of the protein allows close packing of secondary structural elements. Topologically, the surface has no indentations that indicate a nexus for molecular interactions. The distribution of polar and apolar groups on the surface of VapG is markedly uneven. One-third of the surface is dominated by exposed apolar side-chains, with no ionisable and only four polar side-chains exposed, giving rise to an expansive flat hydrophobic surface. Other surface regions are more polar, especially on or near the α-helix and a belt around the centre of the β-barrel. Possible functional significance of these recent structures is discussed. PMID:25746683

  15. Structural characterisation of the virulence-associated protein VapG from the horse pathogen Rhodococcus equi.

    PubMed

    Okoko, Tebekeme; Blagova, Elena V; Whittingham, Jean L; Dover, Lynn G; Wilkinson, Anthony J

    2015-08-31

    Virulence and host range in Rhodococcus equi depends on the variable pathogenicity island of their virulence plasmids. Notable gene products are a family of small secreted virulence-associated proteins (Vaps) that are critical to intramacrophagic proliferation. Equine-adapted strains, which cause severe pyogranulomatous pneumonia in foals, produce a cell-associated VapA that is necessary for virulence, alongside five other secreted homologues. In the absence of biochemical insight, attention has turned to the structures of these proteins to develop a functional hypothesis. Recent studies have described crystal structures for VapD and a truncate of the VapA orthologue of porcine-adapted strains, VapB. Here, we crystallised the full-length VapG and determined its structure by molecular replacement. Electron density corresponding to the N-terminal domain was not visible suggesting that it is disordered. The protein core adopted a compact elliptical, anti-parallel β-barrel fold with β1-β2-β3-β8-β5-β6-β7-β4 topology decorated by a single peripheral α-helix unique to this family. The high glycine content of the protein allows close packing of secondary structural elements. Topologically, the surface has no indentations that indicate a nexus for molecular interactions. The distribution of polar and apolar groups on the surface of VapG is markedly uneven. One-third of the surface is dominated by exposed apolar side-chains, with no ionisable and only four polar side-chains exposed, giving rise to an expansive flat hydrophobic surface. Other surface regions are more polar, especially on or near the α-helix and a belt around the centre of the β-barrel. Possible functional significance of these recent structures is discussed.

  16. Structure of Rhodococcus equi virulence-associated protein B (VapB) reveals an eight-stranded antiparallel β-barrel consisting of two Greek-key motifs

    SciTech Connect

    Geerds, Christina; Wohlmann, Jens; Haas, Albert; Niemann, Hartmut H.

    2014-06-18

    The structure of VapB, a member of the Vap protein family that is involved in virulence of the bacterial pathogen R. equi, was determined by SAD phasing and reveals an eight-stranded antiparallel β-barrel similar to avidin, suggestive of a binding function. Made up of two Greek-key motifs, the topology of VapB is unusual or even unique. Members of the virulence-associated protein (Vap) family from the pathogen Rhodococcus equi regulate virulence in an unknown manner. They do not share recognizable sequence homology with any protein of known structure. VapB and VapA are normally associated with isolates from pigs and horses, respectively. To contribute to a molecular understanding of Vap function, the crystal structure of a protease-resistant VapB fragment was determined at 1.4 Å resolution. The structure was solved by SAD phasing employing the anomalous signal of one endogenous S atom and two bound Co ions with low occupancy. VapB is an eight-stranded antiparallel β-barrel with a single helix. Structural similarity to avidins suggests a potential binding function. Unlike other eight- or ten-stranded β-barrels found in avidins, bacterial outer membrane proteins, fatty-acid-binding proteins and lysozyme inhibitors, Vaps do not have a next-neighbour arrangement but consist of two Greek-key motifs with strand order 41238567, suggesting an unusual or even unique topology.

  17. Comparison of equi-minimum alveolar concentration of sevoflurane and isoflurane on bispectral index values during both wash in and wash out phases: A prospective randomised study.

    PubMed

    Gupta, Madhu; Shri, Iti; Sakia, Prashant; Govil, Deepika

    2015-02-01

    At equal minimum alveolar concentration (MAC), volatile agents may produce different bispectral index (BIS) values especially at low BIS levels when the effect is volatile agent specific. The present study was performed to compare the BIS values produced by sevoflurane and isoflurane at equal MAC and thereby assessing which is a better hypnotic agent. Sixty American Society of Anaesthesiologists I and II patients undergoing elective mastoidectomy were divided into groups receiving either isoflurane or sevoflurane, and at equi-MAC. BIS value was measured during both wash in and wash out phase, keeping other parameters same. Statistical analysis was performed using the Friedman two-way analysis and Mann-Whitney U-test. A P < 0.05 was considered significant. BIS value was significantly lower with sevoflurane at all MAC values as compared to isoflurane, except in the beginning and at MAC awake. However, both the drugs proved to be cardiostable. At equi-MAC sevoflurane produces lower BIS values during wash in as well as wash out phase as compared to isoflurane, reflecting probably an agent specific effect and a deficiency in BIS algorithm for certain agents and their interplay.

  18. Comparison of equi-minimum alveolar concentration of sevoflurane and isoflurane on bispectral index values during both wash in and wash out phases: A prospective randomised study

    PubMed Central

    Gupta, Madhu; Shri, Iti; Sakia, Prashant; Govil, Deepika

    2015-01-01

    Background and Aims: At equal minimum alveolar concentration (MAC), volatile agents may produce different bispectral index (BIS) values especially at low BIS levels when the effect is volatile agent specific. The present study was performed to compare the BIS values produced by sevoflurane and isoflurane at equal MAC and thereby assessing which is a better hypnotic agent. Methods: Sixty American Society of Anaesthesiologists I and II patients undergoing elective mastoidectomy were divided into groups receiving either isoflurane or sevoflurane, and at equi-MAC. BIS value was measured during both wash in and wash out phase, keeping other parameters same. Statistical analysis was performed using the Friedman two-way analysis and Mann-Whitney U-test. A P < 0.05 was considered significant. Results: BIS value was significantly lower with sevoflurane at all MAC values as compared to isoflurane, except in the beginning and at MAC awake. However, both the drugs proved to be cardiostable. Conclusion: At equi-MAC sevoflurane produces lower BIS values during wash in as well as wash out phase as compared to isoflurane, reflecting probably an agent specific effect and a deficiency in BIS algorithm for certain agents and their interplay. PMID:25788739

  19. Analysis of anamnestic immune responses in adult horses and priming in neonates induced by a DNA vaccine expressing the vapA gene of Rhodococcus equi.

    PubMed

    Lopez, A Marianela; Hines, Melissa T; Palmer, Guy H; Knowles, Donald P; Alperin, Debra C; Hines, Stephen A

    2003-09-08

    Rhodococcus equi remains one of the most important pathogens of early life in horses, yet conventional vaccines to prevent rhodococcal pneumonia have not been successful. DNA vaccination offers an alternative to conventional vaccines with specific advantages for immunization of neonates. We developed a DNA vaccine expressing the vapA gene (pVR1055vapA) that induced an anamnestic response characterized by virulence associated protein A (VapA)-specific IgG antibodies in sera and bronchoalveolar lavage fluid (BALF) as well as VapA-specific proliferation of pulmonary lymphocytes when tested in adult ponies. In contrast, none of the adults receiving the control plasmid responded. To determine if pVR1055vapA induced VapA-specific responses in the foal, the targeted age group for vaccination against R. equi, 10 naïve foals were randomly assigned at birth to two groups of five. At 8-15 days of age (day 1), foals were vaccinated by intranasal and intradermal (i.d.) routes with either pVR1055vapA or the negative control pVR1055vapA_rev. All foals were DNA boosted at day 14 and protein boosted at day 30 with either recombinant VapA or recombinant CAT (control group). Prior to the protein boost, neither group developed VapA-specific immune responses. However, at day 45, two of the VR1055vapA-vaccinated foals had increased titers of VapA-specific IgGb, IgM and IgGa in the sera, and IgG in the BALF. The induction of the opsonizing isotypes IgGa and IgGb has been previously shown to be associated with protection against R. equi. No VapA-specific immune responses were detected in the control group. This study indicates that the DNA vaccine effectively stimulates anamnestic systemic and pulmonary immune responses in adult horses. The results in foals suggest that the DNA vaccine also primed a subset of immunized neonates. These data support further development and modification to produce a DNA vaccine to more effectively prime neonatal foals.

  20. Short report: Identification of virulence-associated plasmids in Rhodococcus equi in humans with and without acquired immunodeficiency syndrome in Brazil.

    PubMed

    Ribeiro, Márcio Garcia; Takai, Shinji; de Vargas, Agueda Castagna; Mattos-Guaraldi, Ana Luiza; Ferreira Camello, Thereza Cristina; Ohno, Ryoko; Okano, Hajime; Silva, Aristeu Vieira da

    2011-09-01

    Virulence of Rhodococcus equi strains from 20 humans in Brazil was investigated by using a polymerase chain reaction to characterize isolates as virulent (VapA), intermediately virulent (VapB), and avirulent. Nine isolates were obtained from human immunodeficiency virus (HIV)-positive patients, six from HIV-negative patients, and five from patients of unknown status. Five isolates were VapB positive, four were VapA positive, and eleven were avirulent. Among the nine isolates from HIV-positive patients, five contained VapB plasmids and two contained VapA plasmids. Five VapB-positive isolates had the type 8 virulence plasmid. Eleven of the patients had a history of contact with livestock and/or a farm environment, and none had contact with pigs.

  1. Transcriptional regulation by VirR and VirS of members of the Rhodococcus equi virulence-associated protein multigene family.

    PubMed

    Kakuda, Tsutomu; Miyazaki, Shiko; Hagiuda, Hirofumi; Takai, Shinji

    2015-08-01

    A virulence plasmid of Rhodococcus equi harbors the vap mutigene family. Here it is shown that transcription of vap gene family members other than vapA (vapD, vapE and vapG) is regulated by temperature and pH and abolished when either virS or virR is deleted. Expression of VirS in the absence of functional VirR was found to increase the transcription of vap genes to the amount expressed in the presence of VirR. These findings suggest that transcription of vap genes is regulated by VirS and that VirR is involved in the mechanism of transcriptional responses to temperature and pH. © 2015 The Societies and Wiley Publishing Asia Pty Ltd.

  2. Effect of Solid Die Equi-Channel Pressing Angle on β-Mg17Al12 Phase Morphology and Mechanical Behavior of AZ80 Mg alloy

    NASA Astrophysics Data System (ADS)

    Palai, Pabitra; Prabhu, N.; Kashyap, B. P.

    2017-02-01

    The effects of die channel angle (Φ) in hot ( 623 K) equi-channel angular pressing (ECAP) on microstructure, and tensile and compressive flow properties of AZ80 Mg alloy were investigated. Two solid ECAP dies, having Φ of (1) dual 60° and 120° in a single die and (2) 90° in another die, were designed for this purpose. Grain refinement with more than 40% reduction in average grain size along with submicron size second-phase β-precipitates was achieved after single-pass ECAP. A great variation in β-Mg17Al12 phase morphology with increasing flow stresses in tension and compression are found with decreasing value of angle Φ. There found an increasing effect on strain to failure with decrease in porosity and second-phase precipitate modification. However, there appears flow asymmetry between tension and compression with the latter exhibiting greater flow stress and strain to failure.

  3. The vapA co-expressed virulence plasmid gene vcgB (orf10) of the intracellular actinomycete Rhodococcus equi.

    PubMed

    Miranda-Casoluengo, Raúl; Miranda-Casoluengo, Aleksandra A; O'Connell, Enda P; Fahey, Ruth J; Boland, Clara A; Vázquez-Boland, Jose A; Meijer, Wim G

    2011-08-01

    The virulence plasmid of the pathogenic actinomycete Rhodococcus equi is essential for proliferation of this pathogen in macrophages and the development of disease. The pathogenicity island of this plasmid encodes a family of virulence-associated proteins (Vap), one of which (VapA) is a virulence factor. This paper describes the vcgAB operon (vapA co-expressed gene), located upstream of the vapA operon. Transcription of the vcgAB operon gave rise to transcripts with a half-life similar to those determined for other virulence plasmid genes (1.8 min). Transcription started at a promoter similar to the vapA promoter, and proceeded through an inefficient terminator into the downstream vcgC gene. In addition, vcgC is also transcribed from a promoter downstream of vcgB. The vcgAB and vapA operons were coordinately regulated by temperature and pH in a synergistic manner. The latter parameter only affected transcription at higher growth temperatures, indicating that temperature is the dominant regulatory signal. Transcription of the vcgAB operon increased 10-fold during the late exponential and stationary growth phases. Transcription was also upregulated during the initial hours following phagocytosis by phagocytic cells. In contrast to vcgA and vcgC, the vcgB gene is conserved in the porcine VapB-encoding plasmid, as well as in pathogenic mycobacteria. The coordinated regulation of vcgB and vapA, transcription of vcgB following phagocytosis and conservation of vcgB in pathogenic mycobacteria indicate a role for vcgB and the vcg genes in the virulence of R. equi.

  4. EquiNox2: A new method to measure NADPH oxidase activity and to study effect of inhibitors and their interactions with the enzyme.

    PubMed

    Derochette, Sandrine; Serteyn, Didier; Mouithys-Mickalad, Ange; Ceusters, Justine; Deby-Dupont, Ginette; Neven, Philippe; Franck, Thierry

    2015-11-01

    Excessive neutrophil stimulation and reactive oxygen species (ROS) production are involved in numerous human or horse pathologies. The modulation of the neutrophil NADPH oxidase (NOX) has a great therapeutic potential since this enzyme produces superoxide anion whose most of the other ROS derive. The measurement of NOX activity by cell-free systems is often used to test potential inhibitors of the enzyme. A major drawback of this technique is the possible interferences between inhibitors and the probe, ferricytochrome c, used to measure the activity. We designed the "EquiNox2", a new pharmacological tool, to determine the direct interaction of potential inhibitors with equine phagocytic NOX and their effect on the enzyme activity or assembly. This method consists in binding the membrane fractions of neutrophils containing flavocytochrome b558 or the entire complex, reconstituted in vitro from membrane and cytosolic fractions of PMNs, onto the wells of a microplate followed by incubation with potential inhibitors or drugs. After incubation, the excess of the drug is simply eliminated or washed prior measuring the activity of the reconstituted complex. This latter step avoid the risk of interference between the inhibitor and the revelation solution and can distinguish if inhibitors, strongly bound or not, could interfere with the assembly of the enzymatic complex or with its activity. The EquiNox2 was validated using diphenyliodonium chloride and Gp91ds-tat, two well-known inhibitors largely described for human NADPH oxidase. The present technique was used to study and understand better the effect of curcumin and its water-soluble derivative, NDS27, on the assembly and activity of NOX. We demonstrated that curcumin and NDS27 can strongly bind to the enzyme and prevents its assembly making these molecules good candidates for the treatment of horse or human pathologies implying an excessive activation of neutrophils.

  5. Validation and evaluation of VapA-specific IgG and IgG subclass enzyme-linked immunosorbent assays (ELISAs) to identify foals with Rhodococcus equi pneumonia.

    PubMed

    Sanz, M G; Oliveira, A F; Loynachan, A; Page, A; Svansson, V; Giguère, S; Horohov, D W

    2016-01-01

    Rhodococcus equi (Rhodococcus hoagii/Prescottella equi) is a common cause of foal pneumonia, but its diagnosis remains a challenge for equine veterinarians. While the VapA-specific (virulence-associated protein A) immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) has low sensitivity and specificity for detecting pneumonic foals, little is known about VapA-specific IgG subclasses. To evaluate the performance of VapA-specific ELISA for IgG and its subclasses IgGa, IgGb and IgG(T) in the early diagnosis of pneumonia caused by R. equi. Assay validation followed by assessment of diagnostic performance using archived samples from animals of known status. Serum samples from exposed (n = 125) and nonexposed adult horses (n = 10) and from experimentally challenged and naturally infected foals were used for ELISA validation. Post mortem and tissue culture records of the last 24 years from the Institute for Experimental Pathology at the University of Iceland in Keldur, Iceland laboratory were evaluated to confirm the absence of R. equi cases in Iceland. The diagnostic performance of VapA-specific IgG and its subclasses was evaluated using banked serum samples from pneumonic (n = 21) and healthy foals (n = 80). To evaluate each IgG assay, a cut-off value was selected based on receiver operating characteristic curve analysis and used to calculate sensitivity and specificity. The intra- and interassay coefficients of variation were calculated for each ELISA. Using sera from Iceland, where R. equi infection has not been reported, the VapA-specific IgG ELISA differentiated exposed from nonexposed horses. When used to identify infected foals, VapA-specific IgG, IgGa and IgGb had no diagnostic value. In contrast, IgG(T) had high sensitivity and specificity. Horses from Iceland are not exposed to VapA(+) R. equi and can serve as negative controls. VapA-specific IgG subclasses, with the exception of IgG(T), are poor predictors of disease. Further

  6. Analyses of lipid rafts, Toll-like receptors 2 and 4, and cytokines in foals vaccinated with Virulence Associated Protein A/CpG oligonucleotide vaccine against Rhodococcus equi.

    PubMed

    Kaur, Navjot; Townsend, Hugh; Lohmann, Katharina; Marques, Fernando; Singh, Baljit

    2013-12-15

    Rhodococcus equi establishes long-term pulmonary infection, survives in phagolysosomes of alveolar macrophages and causes pneumonia in foals. The failure of the foal to clear R. equi bacteria is believed to be due to its inability to produce IFN-γ and defects in Toll-like receptor(TLR) signaling. Lipid rafts sequester immune receptors such as TLRs and facilitate efficient cell signaling and therefore, a deficiency in accumulation of receptors in lipid rafts may result in failure to activate. We tested whether a Virulence Associated Protein A (VapA)/CpG vaccine against R. equi would impact the production of IL-10, IFN-γ and TNF-α in lung tissue and fluid samples, alter expression of TLR2 and TLR4 and alter their association with the lipid rafts in broncho-alveolar lavage (BAL) cells. Eight foals, 1–6 days of age, were vaccinated against R. equi followed by a booster at day 14 and challenged with R. equi (5 x 10(6) CFU/ml;10 ml) on day 28. This group was termed "vaccinated pre-challenge" before the infection and "vaccinated post-challenge" after the infection. A second group of foals (n = 7) was not vaccinated but challenged with R. equi on day 28 of the study. This group was termed "non-vaccinated pre-challenge" and after infection with R. equi was named "non-vaccinated post-challenged. We report adaptation of previous protocols to isolate plasma membrane fractions from BAL cells and identification of lipid raft fractions based on the presence of flotillin-1 and GM-1 and absence of transferrin receptor. TLR2 and TLR4 were restricted to plasma membrane fractions 7–9 of alveolar cells collected from vaccinated foals before and after the challenge. Western blots showed that vaccinated post-challenge foals had higher expression of TLR2 in their lung tissues compared to non-vaccinated pre-challenge foals. TNF- concentration was higher in BAL fluid collected from the vaccinated compared to the non-vaccinated foals on day 28. Lung tissue extracts collected on day 49

  7. Prevalence of Theileria equi and Babesia caballi as well as the identification of associated ticks in sympatric Grevy's zebras (Equus grevyi) and donkeys (Equus africanus asinus) in northern Kenya.

    PubMed

    Hawkins, Elaine; Kock, Richard; McKeever, Declan; Gakuya, Francis; Musyoki, Charles; Chege, Stephen M; Mutinda, Mathew; Kariuki, Edward; Davidson, Zeke; Low, Belinda; Skilton, Robert A; Njahira, Moses N; Wamalwa, Mark; Maina, Elsie

    2015-01-01

    The role of equine piroplasmosis as a factor in the population decline of the Grevy's zebra is not known. We determined the prevalence of Babesia caballi and Theileria equi in cograzing Grevy's zebras (Equus grevyi) and donkeys (Equus africanus asinus) in northern Kenya and identified the associated tick vectors. Blood samples were taken from 71 donkeys and 16 Grevy's zebras from March to May 2011. A nested PCR reaction using 18s ribosomal (r)RNA primers on 87 blood spots showed 72% (51/71; 95% confidence interval [CI] 60.4-81.0%) of donkeys and 100% (16/16; 95% CI, 77.3-100%) of Grevy's zebras were T. equi positive. No samples were positive for B. caballi. Sequence comparison using the National Center for Biotechnology Information's basic local alignment search tool identified homologous 18s rRNA sequences with a global geographic spread. The T. equi-derived sequences were evaluated using Bayesian approaches with independent Metropolis-coupled Markov chain Monte Carlo runs. The sequences clustered with those found in Sudan, Croatia, Mongolia, and the US, with statistical support greater than 80% for the two main clades. Hyalomma tick species were found on both donkeys and Grevy's zebras, whereas Rhipicephalus pulchellus was found exclusively on Grevy's zebras and Hyalomma marginatum rupfipes on donkeys. The prevalence of T. equi was 100% in Grevy's zebras and 72% in donkeys with common tick vectors identified. Our results suggest that donkeys and Grevy's zebras can be asymptomatic carriers and that piroplasmosis is endemic in the study area.

  8. Failure of a VapA/CpG oligodeoxynucleotide vaccine to protect foals against experimental Rhocococcus equi pneumonia despite induction of VapA-specific antibody and interferon-γ response

    PubMed Central

    Lohmann, Katharina L.; Lopez, A. Marianela; Manning, Stephen T.; Marques, Fernando J.; Brownlie, Robert; Allen, Andrew L.; Sangster, Anna E.; Mutwiri, George; Gerdts, Volker; Potter, Andrew; Townsend, Hugh G.G.

    2013-01-01

    We evaluated the immunogenic and protective potential of a recombinant VapA/CpG oligodeoxynucleotide (ODN) 2395 vaccine in neonatal foals undergoing experimental Rhodococcus equi challenge. Foals (n = 8) were vaccinated by intramuscular injection on days 1 and 15 of the study; control foals (n = 7) received a phosphate-buffered saline (PBS) solution. All foals were challenged by intrabronchial administration of 5 × 106R. equi 103+ on day 29. Bronchoalveolar lavages were done on days 15, 29, and 36 and total cell count, differential cell count, rVapA-stimulated cell proliferation and interferon (IFN)-γ mRNA expression determined. Clinical examination, complete blood (cell) counts, serology for VapA-specific antibodies, and culture of nasal and fecal swabs were done on days 1, 15, 29, 36, 43, and 50. Foals were humanely euthanized on day 50 and severity of pneumonia scored on a 4-point scale. Vaccination resulted in a significant increase in VapA-specific immunoglobulin (Ig) production, with total IgG and IgG(T) being increased by day 15. Expression of VapA-specific IFN-γ mRNA by BAL cells was increased in the vaccinated foals following challenge. Postmortem lung severity scores did not differ between groups. Two foals shed virulent R. equi in feces; however, real-time polymerase chain reaction (PCR) revealed the isolates to be different from the challenge strain. PMID:24101791

  9. Diversity of Streptococcus equi subsp. zooepidemicus strains isolated from the Spanish sheep and goat population and the identification, function and prevalence of a novel arbutin utilisation system.

    PubMed

    Steward, Karen F; Robinson, Carl; Holden, Matthew T G; Harris, Simon R; Ros, Ana Fernández; Pérez, Gema Chacón; Baselga, Rafael; Waller, Andrew S

    2017-08-01

    The zoonotic bacterium Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is a diverse, opportunistic pathogen that can cause mastitis in dairy sheep and goats. We used multilocus sequence typing (MLST) to define the genetic diversity of 60 isolates of S. zooepidemicus, which were recovered from sheep and goats in Spain between 2003 and 2010. We identify a novel clonal complex based on sequence type (ST), ST-236, which accounted for 39 of the 60 isolates. A representative ST-236 strain, S. zooepidemicus strain C7 (SzC7), was sequenced and interrogated for the presence of novel nutritional uptake or utilisation systems, the acquisition of which have previously been shown to be important for environmental adaptation in other streptococcal pathogens. A novel phosphoenolpyruvate sugar phosphotransferase system (PTS), which enabled the utilisation of arbutin, was identified. Functionality of the PTS was confirmed following deletion of the PTS from SzC7. Arbutin is found in multiple animal foodstuffs and we propose that the ability to utilise arbutin may have conferred a selective advantage to strains infecting animals, the diet of which contains this sugar. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Structure of Rhodococcus equi virulence-associated protein B (VapB) reveals an eight-stranded antiparallel β-barrel consisting of two Greek-key motifs.

    PubMed

    Geerds, Christina; Wohlmann, Jens; Haas, Albert; Niemann, Hartmut H

    2014-07-01

    Members of the virulence-associated protein (Vap) family from the pathogen Rhodococcus equi regulate virulence in an unknown manner. They do not share recognizable sequence homology with any protein of known structure. VapB and VapA are normally associated with isolates from pigs and horses, respectively. To contribute to a molecular understanding of Vap function, the crystal structure of a protease-resistant VapB fragment was determined at 1.4 Å resolution. The structure was solved by SAD phasing employing the anomalous signal of one endogenous S atom and two bound Co ions with low occupancy. VapB is an eight-stranded antiparallel β-barrel with a single helix. Structural similarity to avidins suggests a potential binding function. Unlike other eight- or ten-stranded β-barrels found in avidins, bacterial outer membrane proteins, fatty-acid-binding proteins and lysozyme inhibitors, Vaps do not have a next-neighbour arrangement but consist of two Greek-key motifs with strand order 41238567, suggesting an unusual or even unique topology.

  11. Policies and Processes for Social Inclusion: Using EquiFrame and EquIPP for Policy Dialogue Comment on "Are Sexual and Reproductive Health Policies Designed for All? Vulnerable Groups in Policy Documents of Four European Countries and Their Involvement in Policy Development".

    PubMed

    MacLachlan, Malcolm; Mannan, Hasheem; Huss, Tessy; Munthali, Alister; Amin, Mutamad

    2015-11-16

    The application of EquiFrame in the analysis of sexual and reproductive health policies by Ivanova et al to a new thematic area, their selection of only some of the Core Concepts of human rights in health service provision and the addition of new vulnerable groups relevant to the purpose of their analysis, are all very welcome developments. We also applaud their application of EquiFrame to policies in countries where it has not previously been used, along with their use of interviews with policy-makers to produce a deeper understanding of policy processes. We argue that clear justification for the inclusion of additional, or replacement of some exiting vulnerable groups within EquiFrame should be accompanied by clear definitions of such groups, along with the evidence-base that justifies their classification as a vulnerable or marginalised group. To illustrate the versatility of EquiFrame, we summarise a range of ways in which it has been used across a number of regions; including a brief Case Study of its use to develop the National Health Policy of Malawi. While EquiFrame focuses on policy content, we preview a new policy analysis tool - Equity and Inclusion in Policy Processes (EquIPP) - which assesses the extent of equity and inclusion in broader policy processes. Together, EquiFrame and EquIPP can be used to help governments and civil society ensure that policies are addressing the much stronger emphasis on social inclusion, now apparent in the Sustainable Development Goals (SDGs).

  12. Structural Definition of Trehalose 6-Monomycolates and Trehalose 6,6'-Dimycolates from the Pathogen Rhodococcus equi by Multiple-Stage Linear Ion-Trap Mass Spectrometry with Electrospray Ionization

    NASA Astrophysics Data System (ADS)

    Hsu, Fong-Fu; Wohlmann, Jens; Turk, John; Haas, Albert

    2011-12-01

    The cell wall of the pathogenic bacterium Rhodococcus equi ( R. equi) contains abundant trehalose monomycolate (TMM) and trehalose dimycolate (TDM), the glycolipids bearing mycolic acids. Here, we describe multiple-stage (MS n ) linear ion-trap (LIT) mass spectrometric approaches toward structural characterization of TMM and TDM desorbed as [M + Alk]+ (Alk = Na, Li) and as [M + X]- (X = CH3CO2, HCO2) ions by electrospray ionization (ESI). Upon MS n ( n = 2, 3, 4) on the [M + Alk]+ or the [M + X]- adduct ions of TMM and TDM, abundant structurally informative fragment ions are readily available, permitting fast assignment of the length of the meromycolate chain and of the α-branch on the mycolyl residues. In this way, structures of TMM and TDM isolated from pathogenic R. equi strain 103 can be determined. Our results indicate that the major TMM and TDM molecules possess 6, and/or 6'-mycolyl groups that consist of mainly C14 and C16 α-branches with meromycolate branches ranging from C18 to C28, similar to the structures of the unbound mycolic acids found in the cell envelope. Up to 60 isobaric isomers varying in chain length of the α-branch and of the meromycolate backbone were observed for some of the TDM species in the mixture. This mass spectrometric approach provides a direct method that affords identification of various TMM and TDM isomers in a mixture of which the complexity of this lipid class has not been previously reported using other analytical methods.

  13. Structural definition of trehalose 6-monomycolates and trehalose 6,6'-dimycolates from the pathogen Rhodococcus equi by multiple-stage linear ion-trap mass spectrometry with electrospray ionization

    PubMed Central

    Hsu, Fong-Fu; Wohlmann, Jens; Turk, John; Haas, Albert

    2014-01-01

    The cell wall of the pathogenic bacterium Rhodococcus equi (R. equi) contains abundant trehalose monomycolate (TMM) and trehalose dimycolate (TDM), the glycolipids bearing mycolic acids. Here, we describe multiple-stage (MSn) linear ion-trap (LIT) mass spectrometric approaches toward structural characterization of TMM and TDM desorbed as [M + Alk]+ (Alk = Na, Li) and as [M + X]- (X = CH3CO2, HCO2) ions by electrospray ionization (ESI). Upon MSn (n=2,3,4) on the [M + Alk]+ or the [M + X]− adduct ions of TMM and TDM, abundant structurally informative fragmentations are readily available, permitting fast assignment of the length of the meromycolate chain and of the α-branch on the mycolyl residues. In this way, structures of TMM and TDM isolated from pathogenic R. equi strain 103 can be determined. Our results indicate that the major TMM and TDM molecules possess 6, and/or 6’-mycolyl groups that consist of mainly C14 and C16 α-branches with meromycolate branches ranging from C18 to C28, similar to the structures of the unbound mycolic acids found in the cell envelope. Up to 60 isobaric isomers varying in chain length of the α-branch and of the meromycolate backbone were observed for some of the TDM species in the mixture. This mass spectrometric approach provides a direct method that affords identification of various TMM and TDM isomers in a mixture of which the complexity of this lipid class has not been previously reported using other analytical methods. PMID:21972013

  14. EMS Student Handbook.

    ERIC Educational Resources Information Center

    Ogle, Patrick

    This student guide is one of a series of self-contained materials for students enrolled in an emergency medical services (EMS) training program. Discussed in the individual sections of the guide are the following topics: the purpose and history of EMS professionals; EMS training, certification and examinations (national and state certification and…

  15. EM International. Volume 1

    SciTech Connect

    Not Available

    1993-07-01

    It is the intent of EM International to describe the Office of Environmental Restoration and Waste Management`s (EM`s) various roles and responsibilities within the international community. Cooperative agreements and programs, descriptions of projects and technologies, and synopses of visits to international sites are all highlighted in this semiannual journal. Focus on EM programs in this issue is on international collaboration in vitrification projects. Technology highlights covers: in situ sealing for contaminated sites; and remote sensors for toxic pollutants. Section on profiles of countries includes: Arctic contamination by the former Soviet Union, and EM activities with Germany--cooperative arrangements.

  16. Leveraging EMS and VPP

    DTIC Science & Technology

    2009-05-01

    Elements of EMS  International Standards Organization ( ISO ) 14001 , Environmental Management Systems  The Key Elements of EMS: - Policy - Planning...wingman-- ON and OFF duty Fully Conforming vs. Fully Implemented  “Fully Conforming”  Meets standards established in ISO 14001  ESOH council...e n c e Every airman looking out for his wingman-- ON and OFF duty EMS & VPP Commonalities Environmental Management System ISO 14001 : 2004 Voluntary

  17. EMS in the pueblos.

    PubMed

    Vigil, M A

    1994-02-01

    Imagine creating a movie by excerpting scenes from "Dances With Wolves," splicing it with footage from "Code 3" or "Emergency Response" and then flavoring the script with the mystery of a Tony Hillerman novel. A film producer would probably find it quite difficult to choreograph a finished product from such a compilation of material. To hundreds of Native American EMS providers, however, such a movie is played out every day in Indian country. And with this movie come some real-life problems, including trauma, which is the number-one cause of premature death among Native Americans. But a high trauma rate is just one of the challenges facing tribal EMS responders. There's also prolonged response and transport, the problems involved in maintaining the unique culture and standard of care, the challenges of tribal EMS administration and EMS education of Native American students, and the unsure future of Native American EMS. Beyond that, there's the fact that EMS is a s unique to each Indian reservation as are the cultures of the native peoples who reside on these lands. Yet while no two systems are alike, most tribal EMS providers face similar challenges.

  18. Deletion of the gene encoding the reductase component of 3-ketosteroid 9α-hydroxylase in Rhodococcus equi USA-18 disrupts sterol catabolism, leading to the accumulation of 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid and 1,4-androstadiene-3,17-dione.

    PubMed

    Yeh, Chin-Hsing; Kuo, Yung-Shun; Chang, Che-Ming; Liu, Wen-Hsiung; Sheu, Meei-Ling; Meng, Menghsiao

    2014-09-09

    The gene encoding the putative reductase component (KshB) of 3-ketosteroid 9α-hydroxylase was cloned from Rhodococcus equi USA-18, a cholesterol oxidase-producing strain formerly named Arthrobacter simplex USA-18, by PCR according to consensus amino acid motifs of several bacterial KshB subunits. Deletion of the gene in R. equi USA-18 by a PCR-targeted gene disruption method resulted in a mutant strain that could accumulate up to 0.58 mg/ml 1,4-androstadiene-3,17-dione (ADD) in the culture medium when 0.2% cholesterol was used as the carbon source, indicating the involvement of the deleted enzyme in 9α-hydroxylation of steroids. In addition, this mutant also accumulated 3-oxo-23,24-bisnorchola-1,4-dien-22-oic acid (Δ1,4-BNC). Because both ADD and Δ1,4-BNC are important intermediates for the synthesis of steroid drugs, this mutant derived from R. equi USA-18 may deserve further investigation for its application potential.

  19. EMS in Mauritius.

    PubMed

    Ramalanjaona, Georges; Brogan, Gerald X

    2009-02-01

    Mauritius lies in the southwest Indian Ocean about 1250 miles from the African coast and 500 miles from Madagascar. Mauritius (estimated population 1,230,602) became independent from the United Kingdom in 1968 and has one of the highest GDP per capita in Africa. Within Mauritius there is a well established EMS system with a single 999 national dispatch system. Ambulances are either publicly or privately owned. Public ambulances are run by the Government (SAMU). Megacare is a private subscriber only ambulance service. The Government has recently invested in new technology such as telemedicine to further enhance the role of EMS on the island. This article describes the current state of EMS in Mauritius and depicts its development in the context of Government effort to decentralise and modernise the healthcare system.

  20. EMS -- Error Message Service

    NASA Astrophysics Data System (ADS)

    Rees, P. C. T.; Chipperfield, A. J.; Draper, P. W.

    This document describes the Error Message Service, EMS, and its use in system software. The purpose of EMS is to provide facilities for constructing and storing error messages for future delivery to the user -- usually via the Starlink Error Reporting System, ERR (see SUN/104). EMS can be regarded as a simplified version of ERR without the binding to any software environment (e.g., for message output or access to the parameter and data systems). The routines in this library conform to the error reporting conventions described in SUN/104. A knowledge of these conventions, and of the ADAM system (see SG/4), is assumed in what follows. This document is intended for Starlink systems programmers and can safely be ignored by applications programmers and users.

  1. Assessment of the safety and immunogenicity of Rhodococcus equi-secreted proteins combined with either a liquid nanoparticle (IMS 3012) or a polymeric (PET GEL A) water-based adjuvant in adult horses and foals--identification of promising new candidate antigens.

    PubMed

    Cauchard, S; Bertrand, F; Barrier-Battut, I; Jacquet, S; Laurentie, M; Barbey, C; Laugier, C; Deville, S; Cauchard, J

    2014-02-15

    Rhodococcus equi is the most common infectious cause of mortality in foals between 1 and 6 months of age. Because of an increase in the number of antibiotic-resistant strains, the optimization of a prophylactic strategy is a key factor in the comprehensive management of R. equi pneumonia. The objectives of this study were to assess the safety and immunogenicity of R. equi-secreted proteins (ReSP) co-administered with either the nanoparticular adjuvant Montanide™ IMS 3012 VG, or a new polymeric adjuvant Montanide™ PET GEL A, and to further investigate the most immunogenic proteins for subsequent immunization/challenge experiments in the development of a vaccine against rhodoccocal pneumonia. The approach involved two phases. The first phase aimed to investigate the safety of vaccination in six adult horses. The second phase aimed to determine the safety and immunogenicity of vaccination in twelve 3-week-old foals. We set out to develop a method based on ultrasound measurements for safety assessment in adult horses in order to evaluate any in situ changes at the injection site, in the skin or the underlying muscle, with quantitative and qualitative data revealing that administration of ReSP combined with the Pet Gel A adjuvant led to an increase in local inflammation, associated with 4- to 7-fold higher levels of anti-R. equi IgGa, IgGb and IgGT, compared to administration of ReSP associated with IMS 3012 adjuvant, but without any impact on animal demeanor. Investigations were then performed in foals with serological and clinical follow-up until 6 months of age. Interestingly, we observed in foals a much lower incidence of adverse local tissue reactions at the injection site than in adult horses, with transient and moderate swelling for the group that received ReSP combined with Pet Gel A. Immunized foals with Pet Gel A adjuvant exhibited a similar response in both IgGa and IgGT levels, but a lower response in IgGb levels, compared to adult horses, with a

  2. National EMS Research Agenda.

    PubMed

    Sayre, M R; White, L J; Brown, L H; McHenry, S D

    2002-01-01

    Now, more than ever before, the spirit of the emergency services professional is recognized by people everywhere. Individuals from every walk of life comprehend the reality of the job these professionals do each day. Placing the safety of others above their own is their acknowledged responsibility. Rescue and treatment of ill and injured patients are their purpose as well as their gratification. The men and women who provide prehospital care are well aware of the unpredictable nature of emergency medical services (EMS). Prehospital care is given when and where it is needed: in urban settings with vertical challenges and gridlock; in rural settings with limited access; in confined spaces; within entrapments; or simply in the street, exposed to the elements. Despite the challenges, EMS professionals rise to the occasion to do their best with the resources available. Despite more than 30 years of dedicated service by thousands of EMS professionals, academic researchers, and public policy makers, the nation's EMS system is treating victims of illness and injury with little or no evidence that the care they provide is optimal. A national investment in the EMS research infrastructure is necessary to overcome obstacles currently impeding the accumulation of essential evidence of the effectiveness of EMS practice. Funding is required to train new researchers and to help them establish their careers. Financial backing is needed to support the development of effective prehospital treatments for the diseases that drive the design of the EMS system, including injury and sudden cardiac arrest. Innovative strategies to make EMS research easier to accomplish in emergency situations must be implemented. Researchers must have access to patient outcome information in order to evaluate and improve prehospital care. New biomedical and technical advances must be evaluated using scientific methodology. Research is the key to maintaining focus on improving the overall health of the

  3. Phase evolution, microstructure and mechanical properties of equi-atomic substituted TiZrHfNiCu and TiZrHfNiCuM (M = Co, Nb) high-entropy alloys

    NASA Astrophysics Data System (ADS)

    Park, Hae Jin; Na, Young Sang; Hong, Sung Hwan; Kim, Jeong Tae; Kim, Young Seok; Lim, Ka Ram; Park, Jin Man; Kim, Ki Buem

    2016-07-01

    In this study, alloys with composition of equi-atomic substituted TiZrHfNiCu, TiZrHfNiCuCo, and TiZrHfNiCuNb high-entropy alloys (HEAs) were produced by suction casting method. The effects of addition elements on phase composition, microstructure and mechanical behaviors of the HEA were studied. The suction casted Ti20Zr20Hf20Ni20Cu20 HEA exhibits single C14 Laves phase (MgZn2-type) with fine homogeneous microstructure. When Co or Nb elements are added, morphologies are slightly modulated toward well-developed dendritic microstructure, phase constitutions are significantly changed from single Laves phase to mixed multi-phases as well as mechanical properties are also altered with increased plasticity and high strength. It is believed that modulated mechanical properties are mainly ascribed to the change of phase constitution and crystalline structure, together with the microstructural characteristics. This clearly reveals that the selection and addition of supplementary elements based on the formation rule for HEAs play an important role on the evolution of phase, microstructural morphology and mechanical properties of Ti20Zr20Hf20Ni20Cu20 HEA.

  4. Characterization of Streptococcus equi subsp. ruminatorum isolated from spotted hyenas (Crocuta crocuta) and plains zebras (Equus burchelli), and identification of a M-like protein (SrM) encoding gene.

    PubMed

    Speck, Stephanie; Höner, Oliver P; Wachter, Bettina; Fickel, Jörns

    2008-04-01

    Thirteen strains of Streptococcus equi subsp. ruminatorum from free-ranging spotted hyenas (Crocuta crocuta) and plains zebras (Equus burchelli) in Tanzania were characterized by biochemical and molecular-biological methods. Although the colony appearance of the S.e. ruminatorum wildlife strains differed from that of the S.e. ruminatorum type strain CECT 5772(T), all biochemical reactions of the wildlife strains were similar to those of the type strain. In addition, all wildlife strains produced hyaluronidase and were capable of hydrolysing arginine, three strains (23%) synthesized acetoin, but only eight strains (62%) produced acid from ribose. rep-PCR indicated that different clones of S.e. ruminatorum were distributed among the hyena and zebra populations in the study area. Identical rep-PCR patterns in hyena and zebra strains suggest that a direct transmission of S.e. ruminatorum between these species may occur. The presence of a M-like protein (SrM) gene was demonstrated in all S.e. ruminatorum strains including the type strain. Sequencing of the M-like protein gene revealed a hypervariable region within the deduced amino acid sequence. Most of the strains clustered with previously described strains based on the hypervariable region of the S.e. zooepidemicus SzP protein. Sequencing also demonstrated that identical SrM protein sequences were shared among S.e. ruminatorum strains from different host species.

  5. The EM Earthquake Precursor

    NASA Astrophysics Data System (ADS)

    Jones, K. B., II; Saxton, P. T.

    2013-12-01

    Many attempts have been made to determine a sound forecasting method regarding earthquakes and warn the public in turn. Presently, the animal kingdom leads the precursor list alluding to a transmission related source. By applying the animal-based model to an electromagnetic (EM) wave model, various hypotheses were formed, but the most interesting one required the use of a magnetometer with a differing design and geometry. To date, numerous, high-end magnetometers have been in use in close proximity to fault zones for potential earthquake forecasting; however, something is still amiss. The problem still resides with what exactly is forecastable and the investigating direction of EM. After the 1989 Loma Prieta Earthquake, American earthquake investigators predetermined magnetometer use and a minimum earthquake magnitude necessary for EM detection. This action was set in motion, due to the extensive damage incurred and public outrage concerning earthquake forecasting; however, the magnetometers employed, grounded or buried, are completely subject to static and electric fields and have yet to correlate to an identifiable precursor. Secondly, there is neither a networked array for finding any epicentral locations, nor have there been any attempts to find even one. This methodology needs dismissal, because it is overly complicated, subject to continuous change, and provides no response time. As for the minimum magnitude threshold, which was set at M5, this is simply higher than what modern technological advances have gained. Detection can now be achieved at approximately M1, which greatly improves forecasting chances. A propagating precursor has now been detected in both the field and laboratory. Field antenna testing conducted outside the NE Texas town of Timpson in February, 2013, detected three strong EM sources along with numerous weaker signals. The antenna had mobility, and observations were noted for recurrence, duration, and frequency response. Next, two

  6. EMS & the DEA.

    PubMed

    Beeson, Jeff; Ayres, Chris

    2010-01-01

    It's clear that EMS medical directors and management staff must be vigilant in their oversight of implementation, administration and monitoring of controlled substances within their agencies to best serve the public and avoid running afoul of investigation and incurring significant penalties. Those potentially affected by the need for individual registrations of both emergency vehicles and central inventory systems should carefully monitor upcoming developments in the interpretation od DEA regulations.

  7. Why do Emergency Medical Services (EMS) Professionals Leave EMS?

    PubMed

    Blau, Gary; Chapman, Susan A

    2016-12-01

    The objective was to determine why Emergency Medical Technician (EMT)-Basics and Paramedics leave the Emergency Medical Services (EMS) workforce. Data were collected through annual surveys of nationally registered EMT-Basics and Paramedics from 1999 to 2008. Survey items dealing with satisfaction with the EMS profession, likelihood of leaving the profession, and likelihood of leaving their EMS job were assessed for both EMT-Basics and Paramedics, along with reasons for leaving the profession. Individuals whose responses indicated that they were not working in EMS were mailed a special exit survey to determine the reasons for leaving EMS. The likelihood of leaving the profession in the next year was low for both EMT-Basics and Paramedics. Although overall satisfaction levels with the profession were high, EMT-Basics were significantly more satisfied than Paramedics. The most important reasons for leaving the profession were choosing to pursue further education and moving to a new location. A desire for better pay and benefits was a significantly more important reason for EMT-Paramedics' exit decisions than for EMT-Basics. Given the anticipated increased demand for EMS professionals in the next decade, continued study of issues associated with retention is strongly recommended. Some specific recommendations and suggestions for promoting retention are provided. Blau G , Chapman SA . Why do Emergency Medical Services (EMS) professionals leave EMS? Prehosp Disaster Med. 2016;31(Suppl. 1):s105-s111.

  8. [Experimental prospective cognitive study on the evaluation of connection of fatigue perception and depression among caregivers and team treating cancer patients receiving oral chemotherapy to systemic chemotherapy. Equi-Car 10 study].

    PubMed

    Barzelloni, Maria Luisa; Mogavero, Anna; Carnicelli, Pietro; Izzo, Rosanna; Crivaro, Valeria

    2012-01-01

    The Equi-Car 10 is an spontaneous, observational study. The objective was to assess the connection of fatigue perception and depression of caregivers, oncological team and oncological patients in oral chemotherapy to intravenous The study recruited 60 patients and 60 caregivers. The study was carried out on patients with lung, breast and colon cancer requiring chemotherapy, and family members were chosen as caregivers. Patients, caregivers and doctors filled in FACT-F and Zung questionnaires at the opening day of the medical record, after three and six cycles of chemotherapy, and three months after the end of the treatment. The Zung depression and Functional Assessment of Cancer therapy-Fatigue (FACT-F) showed: (a) on patients treated with intravenous therapy showed significantly higher levels of fatigue and depression than those of patients treated with oral therapy, even when the latter were treated for advanced disease; (b) on caregivers of patients treated with systemically administered therapies showed elevated levels of Fatigue and depression which proved to be significantly higher than those of caregivers of patients treated with oral therapies, even when treated for advanced disease; (c) on three oncologist has highlighted low levels of Fatigue and depression, for both types of patients, higher for patients with systemic therapy related to the management of side effects. The results showed that cancer patients and caregivers have high levels of fatigue and depression both related to the disease stage and to the mode of drug administration. It is necessary to provide cancer patients and caregivers with appropriate psychological support and preventive programs for secondary and relapse prevention.

  9. Identified EM Earthquake Precursors

    NASA Astrophysics Data System (ADS)

    Jones, Kenneth, II; Saxton, Patrick

    2014-05-01

    Many attempts have been made to determine a sound forecasting method regarding earthquakes and warn the public in turn. Presently, the animal kingdom leads the precursor list alluding to a transmission related source. By applying the animal-based model to an electromagnetic (EM) wave model, various hypotheses were formed, but the most interesting one required the use of a magnetometer with a differing design and geometry. To date, numerous, high-end magnetometers have been in use in close proximity to fault zones for potential earthquake forecasting; however, something is still amiss. The problem still resides with what exactly is forecastable and the investigating direction of EM. After a number of custom rock experiments, two hypotheses were formed which could answer the EM wave model. The first hypothesis concerned a sufficient and continuous electron movement either by surface or penetrative flow, and the second regarded a novel approach to radio transmission. Electron flow along fracture surfaces was determined to be inadequate in creating strong EM fields, because rock has a very high electrical resistance making it a high quality insulator. Penetrative flow could not be corroborated as well, because it was discovered that rock was absorbing and confining electrons to a very thin skin depth. Radio wave transmission and detection worked with every single test administered. This hypothesis was reviewed for propagating, long-wave generation with sufficient amplitude, and the capability of penetrating solid rock. Additionally, fracture spaces, either air or ion-filled, can facilitate this concept from great depths and allow for surficial detection. A few propagating precursor signals have been detected in the field occurring with associated phases using custom-built loop antennae. Field testing was conducted in Southern California from 2006-2011, and outside the NE Texas town of Timpson in February, 2013. The antennae have mobility and observations were noted for

  10. EPA LABORATORIES IMPLEMENT EMS PROGRAM

    EPA Science Inventory

    This paper highlights the breadth and magnitude of carrying out an effective Environmental Management System (EMS) program at the U.S. EPA's research and development laboratories. Federal research laboratories have unique operating challenges compared to more centralized industr...

  11. Busca de estruturas em grandes escalas em altos redshifts

    NASA Astrophysics Data System (ADS)

    Boris, N. V.; Sodrã©, L., Jr.; Cypriano, E.

    2003-08-01

    A busca por estruturas em grandes escalas (aglomerados de galáxias, por exemplo) é um ativo tópico de pesquisas hoje em dia, pois a detecção de um único aglomerado em altos redshifts pode por vínculos fortes sobre os modelos cosmológicos. Neste projeto estamos fazendo uma busca de estruturas distantes em campos contendo pares de quasares próximos entre si em z Â3 0.9. Os pares de quasares foram extraídos do catálogo de Véron-Cetty & Véron (2001) e estão sendo observados com os telescópios: 2,2m da University of Hawaii (UH), 2,5m do Observatório de Las Campanas e com o GEMINI. Apresentamos aqui a análise preliminar de um par de quasares observado nos filtros i'(7800 Å) e z'(9500 Å) com o GEMINI. A cor (i'-z') mostrou-se útil para detectar objetos "early-type" em redshifts menores que 1.1. No estudo do par 131046+0006/J131055+0008, com redshift ~ 0.9, o uso deste método possibilitou a detecção de sete objetos candidatos a galáxias "early-type". Num mapa da distribuição projetada dos objetos para 22 < i' < 25 observou-se que estas galáxias estão localizadas próximas a um dos quasares e há indícios de que estejam aglomeradas dentro de um área de ~ 6 arcmin2. Se esse for o caso, estes objetos seriam membros de uma estrutura em grande escala. Um outro argumento em favor dessa hipótese é que eles obedecem uma relação do tipo Kormendy (raio equivalente X brilho superficial dentro desse raio), como a apresentada pelas galáxias elípticas em z = 0.

  12. The European Mobile System (EMS)

    NASA Technical Reports Server (NTRS)

    Jongejans, A.; Rogard, R.; Mistretta, I.; Ananasso, F.

    1993-01-01

    The European Space Agency is presently procuring an L band payload in order to promote a regional European L band system coping with the specific needs of the European market. The payload, and the two communications systems to be supported, are described below. The potential market for EMS in Europe is discussed.

  13. Cosmic muon induced EM showers in NO$\

    SciTech Connect

    Yadav, Nitin; Duyang, Hongyue; Shanahan, Peter; Mishra, Sanjib; Bhuyan, Bipul

    2016-11-15

    Here, the NuMI Off-Axis ve Appearance (NOvA) experiment is a ne appearance neutrino oscillation experiment at Fermilab. It identifies the ne signal from the electromagnetic (EM) showers induced by the electrons in the final state of neutrino interactions. Cosmic muon induced EM showers, dominated by bremsstrahlung, are abundant in NOvA far detector. We use the Cosmic Muon- Removal technique to get pure EM shower sample from bremsstrahlung muons in data. We also use Cosmic muon decay in flight EM showers which are highly pure EM showers.The large Cosmic-EM sample can be used, as data driven method, to characterize the EM shower signature and provides valuable checks of the simulation, reconstruction, particle identification algorithm, and calibration across the NOvA detector.

  14. Correlation of the NBME advanced clinical examination in EM and the national EM M4 exams.

    PubMed

    Hiller, Katherine; Miller, Emily S; Lawson, Luan; Wald, David; Beeson, Michael; Heitz, Corey; Morrissey, Thomas; House, Joseph; Poznanski, Stacey

    2015-01-01

    Since 2011 two online, validated exams for fourth-year emergency medicine (EM) students have been available (National EM M4 Exams). In 2013 the National Board of Medical Examiners offered the Advanced Clinical Examination in Emergency Medicine (EM-ACE). All of these exams are now in widespread use; however, there are no data on how they correlate. This study evaluated the correlation between the EM-ACE exam and the National EM M4 Exams. From May 2013 to April 2014 the EM-ACE and one version of the EM M4 exam were administered sequentially to fourth-year EM students at five U.S. medical schools. Data collected included institution, gross and scaled scores and version of the EM M4 exam. We performed Pearson's correlation and random effects linear regression. 305 students took the EM-ACE and versions 1 (V1) or 2 (V2) of the EM M4 exams (281 and 24, respectively) [corrected].The mean percent correct for the exams were as follows: EM-ACE 74.9 (SD-9.82), V1 83.0 (SD-6.39), V2 78.5 (SD-7.70) [corrected]. Pearson's correlation coefficient for the V1/EM-ACE was 0.53 (0.43 scaled) and for the V2/EM-ACE was 0.58 (0.41 scaled) [corrected]. The coefficient of determination for V1/ EM-ACE was 0.73 and for V2/EM-ACE 0.71 (0.65 and .49 for scaled scores) [ERRATUM]. The R-squared values were 0.28 and 0.30 (0.18 and 0.13 scaled), respectively [corrected]. There was significant cluster effect by institution. There was moderate positive correlation of student scores on the EM-ACE exam and the National EM M4 Exams.

  15. Complaints against an EMS system.

    PubMed

    Colwell, Christopher B; Pons, Peter T; Pi, Randy

    2003-11-01

    Complaints against Emergency Medical Services (EMS) agencies represent a concerning and potentially time-consuming problem for all involved in the delivery of prehospital emergency medical care. The objective of this study was to identify the source of complaints against an EMS system to help focus quality and performance improvement and customer service efforts. We conducted a retrospective review of complaints filed against a busy urban EMS agency over a 6-year period. All complaints were included, totaled by season and by year, and categorized by originator and nature of the complaint. A total of 286 complaints were registered during the 6-year period, with an average of 48 per year and 9.3 per 10,000 responses. The most common originators of complaints were patients (53%) followed by medical personnel (19%) and family members or friends (12%). Rude behavior accounted for 23% of the complaints registered, followed by technical skills (20%), transport problems (18%), and loss of belongings (13%). The identification of areas of dissatisfaction will allow focused quality and performance improvement programs directed at customer service and risk management.

  16. Cosmic muon induced EM showers in NO$$\

    DOE PAGES

    Yadav, Nitin; Duyang, Hongyue; Shanahan, Peter; ...

    2016-11-15

    Here, the NuMI Off-Axis ve Appearance (NOvA) experiment is a ne appearance neutrino oscillation experiment at Fermilab. It identifies the ne signal from the electromagnetic (EM) showers induced by the electrons in the final state of neutrino interactions. Cosmic muon induced EM showers, dominated by bremsstrahlung, are abundant in NOvA far detector. We use the Cosmic Muon- Removal technique to get pure EM shower sample from bremsstrahlung muons in data. We also use Cosmic muon decay in flight EM showers which are highly pure EM showers.The large Cosmic-EM sample can be used, as data driven method, to characterize the EMmore » shower signature and provides valuable checks of the simulation, reconstruction, particle identification algorithm, and calibration across the NOvA detector.« less

  17. The Empathic Operating System (emOS)

    DTIC Science & Technology

    2016-06-15

    The Empathic Operating System ( emOS ) Physiological measurements have typically been limited to expensive and cumbersome clinical research equipment...Report: The Empathic Operating System ( emOS ) Report Title Physiological measurements have typically been limited to expensive and cumbersome clinical...C-0043 Proposal number: 62850-LS-DRP Project title: “The Empathic Operating System ( emOS )” Contract performance period: January 1, 2016 – April 7

  18. DOE/EM Criticality Safety Needs Assessment

    SciTech Connect

    Westfall, Robert Michael; Hopper, Calvin Mitchell

    2011-02-01

    The issue of nuclear criticality safety (NCS) in Department of Energy Environmental Management (DOE/EM) fissionable material operations presents challenges because of the large quantities of material present in the facilities and equipment that are committed to storage and/or material conditioning and dispositioning processes. Given the uncertainty associated with the material and conditions for many DOE/EM fissionable material operations, ensuring safety while maintaining operational efficiency requires the application of the most-effective criticality safety practices. In turn, more-efficient implementation of these practices can be achieved if the best NCS technologies are utilized. In 2002, DOE/EM-1 commissioned a survey of criticality safety technical needs at the major EM sites. These needs were documented in the report Analysis of Nuclear Criticality Safety Technology Supporting the Environmental Management Program, issued May 2002. Subsequent to this study, EM safety management personnel made a commitment to applying the best and latest criticality safety technology, as described by the DOE Nuclear Criticality Safety Program (NCSP). Over the past 7 years, this commitment has enabled the transfer of several new technologies to EM operations. In 2008, it was decided to broaden the basis of the EM NCS needs assessment to include not only current needs for technologies but also NCS operational areas with potential for improvements in controls, analysis, and regulations. A series of NCS workshops has been conducted over the past years, and needs have been identified and addressed by EM staff and contractor personnel. These workshops were organized and conducted by the EM Criticality Safety Program Manager with administrative and technical support by staff at Oak Ridge National Laboratory (ORNL). This report records the progress made in identifying the needs, determining the approaches for addressing these needs, and assimilating new NCS technologies into EM

  19. EMS provider determinations of necessity for transport and reimbursement for EMS response, medical care, and transport: combined resource document for the National Association of EMS Physicians position statements.

    PubMed

    Millin, Michael G; Brown, Lawrence H; Schwartz, Brian

    2011-01-01

    With increasing demands for emergency medical services (EMS), many EMS jurisdictions are utilizing EMS provider-initiated nontransport policies as a method to offload potentially nonemergent patients from the EMS system. EMS provider determination of medical necessity, resulting in nontransport of patients, has the potential to avert unnecessary emergency department visits. However, EMS systems that utilize these policies must have additional education for the providers, a quality improvement process, and active physician oversight. In addition, EMS provider determination of nontransport for a specific situation should be supported by evidence in the peer-reviewed literature that the practice is safe. Further, EMS systems that do not utilize these programs should not be financially penalized. Payment for EMS services should be based on the prudent layperson standard. EMS systems that do utilize nontransport policies should be appropriately reimbursed, as this represents potential cost savings to the health care system.

  20. School Budget Hold'em Facilitator's Guide

    ERIC Educational Resources Information Center

    Education Resource Strategies, 2012

    2012-01-01

    "School Budget Hold'em" is a game designed to help school districts rethink their budgeting process. It evolved out of Education Resource Strategies' (ERS) experience working with large urban districts around the country. "School Budget Hold'em" offers a completely new approach--one that can turn the budgeting process into a long-term visioning…

  1. EM Cep: The Be Star

    NASA Astrophysics Data System (ADS)

    Kochiashvili, N.; Kochiashvili, I.; Natsvlishvili, R.; Vardosanidze, M.; Beradze, S.

    2017-07-01

    On the basis of UBVR photometric data, obtained in the Abastumani Observatory during 1991-1999, very interesting and unusual flare of EM Cep has been revealed. Duration of the flare was over two hours. We estimated the percentage of brightness increase during the flare and brightness decrease of the corresponding anti- flare and the minimum amount of the lost mass during this event. We have solved the light curves of the star using the Wilson-Devinney code. But the resulting fraction of calculated brightness of the companion star was not in accordance with spectral data. Then we decided to check the idea of a pulsating single star using new spectral data. Together with our Buyrakan colleagues we obtained and analyzed spectra of the star. We could not find spectral lines of a companion star or any traces of the radial velocities using this data. Hence, we concluded that we need the higher resolution spectra for final resolution of the matter. On the basis of the latest spectral data of Bulgarian astronomers they concluded that EM Cep is a single star. This makes it possible to suggest, that the question of stellar pulsation could be solved using additional photometric observations.

  2. The EM algorithm in medical imaging.

    PubMed

    Kay, J

    1997-03-01

    This article outlines the statistical developments that have taken place in the use of the EM algorithm in emission and transmission tomography during the past decade or so. We discuss the statistical aspects of the modelling of the projection data for both the emission and transmission cases and define the relevant probability models. This leads to the use of the method of maximum likelihood as a means of estimating the relevant unknown parameters within a given region of a patient's body and to the use of the EM algorithm to compute the reconstruction. Various different types of EM algorithm are discussed, including the SAGE algorithms of Fessler and Hero. The limitations of the EM algorithm, per se, are covered and the need for regularization is stressed. A number of different methods for penalizing the likelihood are described and a number of algorithms for the computation of the penalized EM reconstruction are discussed.

  3. Side mounted EMS for aluminium scrap melters

    SciTech Connect

    Eidem, M.; Tallbaeck, G.; Hanley, P.J.

    1996-10-01

    Normally the electromagnetic stirrer (EMS) is placed below the furnace. However it has recently been found that the EMS can also be placed at the side of the furnace, still giving good stirring. This makes it possible to install EMS on most existing furnaces. The side-mounted EMS is compared with the standard bottom-mounted stirrer with respect to installation, melting time and flow pattern in the melt. The major conclusion is that a side-mounted EMS is practical and will give about as good a performance as the bottom-mounted. Melting time estimates are based upon 3-D fluid flow and heat transfer predictions in combination with a simplified scrap melting theory. Predicted melting times are in fair agreement with operational data for mechanically stirred and electromagnetically bottom stirred furnaces.

  4. EM International, July 1994, Volume 2

    SciTech Connect

    Not Available

    1994-10-01

    The Office of Environmental Management (EM) at the Department of Energy (DOE) is seeking out and leveraging foreign technology, data, and resources in keeping with EM`s mandate to protect public health and the environment through the safe and cost-effective remediation of the Department`s nuclear weapons sites. EM works closely with foreign governments, industry, and universities to obtain innovative environmental technologies, scientific and engineering expertise, and operations experience that will support EM`s objectives. Where appropriate, these international resources are used to manage the more urgent risks at our sites, secure a safe workplace, help build consensus on critical issues, and strengthen our technology development program. Through international agreements EM engages in cooperative exchange of information, technology, and individuals. Currently, we are managing agreements with a dozen countries in Europe, Latin America, and Asia. These agreements focus on environmental restoration, waste management, transportation of radioactive wastes, and decontamination and decommissioning. This publication contains the following articles: in situ remediation integrated program; in-situ characterization and inspection of tanks; multimedia environmental pollutant assessment system (MEPAS); LLNL wet oxidation -- AEA technology. Besides these articles, this publication covers: EU activities with Russia; technology transfer activities; and international organization activities.

  5. Project X RFQ EM Design

    SciTech Connect

    Romanov, Gennady; Hoff, Matthew; Li, Derun; Staples, John; Virostek, Steve; /LBNL

    2012-05-09

    Project X is a proposed multi-MW proton facility at Fermi National Accelerator Laboratory (FNAL). The Project X front-end would consist of an H- ion source, a low-energy beam transport (LEBT), a CW 162.5 MHz radio-frequency quadrupole (RFQ) accelerator, and a medium-energy beam transport (MEBT). Lawrence Berkeley National Laboratory (LBNL) and FNAL collaboration is currently developing the designs for various components in the Project X front end. This paper reports the detailed EM design of the CW 162.5 MHz RFQ that provides bunching of the 1-10 mA H- beam with acceleration from 30 keV to 2.1 MeV.

  6. NEIC Environmental Management System (EMS) Policy

    EPA Pesticide Factsheets

    National Enforcement Investigations Center (NEIC) Environmental Management System (EMS) Policy. Identification and management of actual and potential environmental impacts of operations and decisions for the purpose of continual improvement of performance

  7. EMS offshore. A new horizon for paramedics.

    PubMed

    Mallard, A S

    1991-10-01

    The difficulty in getting medical aid to offshore drilling platforms can be a source of life-threatening delays. Recently, some companies have charted new waters by actually stationing EMS crews on their rigs.

  8. How good can cryo-EM become?

    SciTech Connect

    Glaeser, Robert M.

    2015-12-30

    The suddenness with which single-particle cryo-electron microscopy (cryo-EM) has emerged as a method for determining high-resolution structures of biological macromolecules invites the questions, how much better can this technology get, and how fast is that likely to happen? While we can rightly celebrate the maturation of cryo-EM as a high-resolution structure-determination tool, I believe there still are many developments to look forward to.

  9. Reassessing training levels for prehospital EMS personnel.

    PubMed

    Briese, G L

    1983-01-01

    One of the major questions confronting prehospital care services today concerns determining the appropriate level of training for EMS personnel that will provide the most cost effective systems. Unfortunately there are no studies which assess this problem. Various communities have modified or expanded the roles of prehospital personnel beyond the traditional training of EMTs and paramedics. Continuing education and skills maintenance are ongoing problems faced by all EMS systems, which have been addressed in various ways by individual locales.

  10. SOFIS FTS EM test results

    NASA Astrophysics Data System (ADS)

    Soucy, Marc-Andre A.; Levesque, Luc E.; Tanii, Jun; Kawashima, Takahiro; Nakajima, Hideaki

    2003-04-01

    The Solar Occultation FTS for Inclined-orbit Satellite (SOFIS) is a solar occultation Fourier transform spectrometer developed by the Ministry of the Environment (MOE) in Japan for the Global Change Observation Mission-A1 (GCOM-A1) satellite. GCOM-A1 will be placed in a 650 km non-sun-synchronous orbit, with an inclination angle of 69 degrees. ABB-Bomem is a sub-contractor of NTSpace (NEC-Toshiba Space) for the design and manufacturing of the FTS Engineering Model of SOFIS. SOFIS measures the vertical profile of the atmospheric constituents with 0.2 cm-1 spectral resolution for the spectral range covering 3-13 μm. The atmospheric vertical resolution of SOFIS is 1 km. The target of SOFIS measurements is a global distribution of O3, HNO3, NO2, N2O, CH4, H2O, CO2, CFC-11, CFC-12, ClONO2, aerosol extinction, atmospheric pressure and temperature. NTSpace in Japan is the prime contractor of SOFIS. The spectrometer is an adapted version of the classical Michelson interferometer using an optimized optical layout and moving retro-reflectors. A solid-state laser diode operating at 1550 nm is used as metrology source of the interferometer. Its highly folded optical design results in a high performance instrument with a compact size. SOFIS FTS implements high performance control techniques to achieve outstanding speed stability of the moving mechanism. This paper describes the test activities of the SOFIS-FTS Engineering Model (EM) and preliminary results. The performances of the FTS are presented in terms of key parameters like signal-to-noise ratio, modulation efficiency and stability. Spectra acquired are shown and test methodology and analyses are presented. Lessons learned during assembly, integration and testing are described as well as improvements planned to be implemented in the Flight Model.

  11. EMS adaptation for climate change

    NASA Astrophysics Data System (ADS)

    Pan, C.; Chang, Y.; Wen, J.; Tsai, M.

    2010-12-01

    The purpose of this study was to find an appropriate scenario of pre-hospital transportation of an emergency medical service (EMS) system for burdensome casualties resulting from extreme climate events. A case of natural catastrophic events in Taiwan, 88 wind-caused disasters, was reviewed and analyzed. A sequential-conveyance method was designed to shorten the casualty transportation time and to promote the efficiency of ambulance services. A proposed mobile emergency medical center was first constructed in a safe area, but nearby the disaster area. The Center consists of professional medical personnel who process the triage of incoming patients and take care of casualties with minor injuries. Ambulances in the Center were ready to sequentially convey the casualties with severer conditions to an assigned hospital that is distant from the disaster area for further treatment. The study suggests that if we could construct a spacious and well-equipped mobile emergency medical center, only a small portion of casualties would need to be transferred to distant hospitals. This would reduce the over-crowding problem in hospital ERs. First-line ambulances only reciprocated between the mobile emergency medical center and the disaster area, saving time and shortening the working distances. Second-line ambulances were highly regulated between the mobile emergency medical center and requested hospitals. The ambulance service of the sequential-conveyance method was found to be more efficient than the conventional method and was concluded to be more profitable and reasonable on paper in adapting to climate change. Therefore, additional practical work should be launched to collect more precise quantitative data.

  12. Filter banks and the EM algorithm

    SciTech Connect

    Mair, B.A.; Carroll, R.B.; Anderson, J.M.M.

    1996-12-31

    In this paper, we present a wavelet based modification of the ML-EM algorithm for reconstructing positron emission tomography images. By using the filter bank implementation of the wavelet transform, this algorithm has the flexibility to incorporate a priori information, while maintaining the same computational complexity as the standard ML-EM algorithm. Thus, it has a significant computational advantage over usual Bayesian methods. It differs from recent wavelet-based Bayesian methods as it achieves {open_quotes}regularization{close_quotes} by an adaptive, wavelet-based method of thresholding which minimizes Stein`s Unbiased Estimate of Risk. The basic method consists of applying Donoho and Johnstone`s SureShrink wavelet denoising of the Poisson data, and then applying the standard ML-EM algorithm to the denoised data. A more elaborate method is discussed in which a wavelet denoising step is inserted after each EM iteration. This technique differs from previous smoothing techniques applied to the ML-EM algorithm since it is able to recover edges in discontinuous images.

  13. Processing of Cryo-EM Movie Data.

    PubMed

    Ripstein, Z A; Rubinstein, J L

    2016-01-01

    Direct detector device (DDD) cameras dramatically enhance the capabilities of electron cryomicroscopy (cryo-EM) due to their improved detective quantum efficiency (DQE) relative to other detectors. DDDs use semiconductor technology that allows micrographs to be recorded as movies rather than integrated individual exposures. Movies from DDDs improve cryo-EM in another, more surprising, way. DDD movies revealed beam-induced specimen movement as a major source of image degradation and provide a way to partially correct the problem by aligning frames or regions of frames to account for this specimen movement. In this chapter, we use a self-consistent mathematical notation to explain, compare, and contrast several of the most popular existing algorithms for computationally correcting specimen movement in DDD movies. We conclude by discussing future developments in algorithms for processing DDD movies that would extend the capabilities of cryo-EM even further. © 2016 Elsevier Inc. All rights reserved.

  14. 7 CFR 1945.35 - Special EM loan training.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... workshop and a test. (c) Objective. The basic objective of this training program is to keep State and... kit, will be used for the EM loan training meetings, and any subsequent EM loan training...

  15. 7 CFR 1945.35 - Special EM loan training.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... workshop and a test. (c) Objective. The basic objective of this training program is to keep State and... kit, will be used for the EM loan training meetings, and any subsequent EM loan training...

  16. Integrating Green Purchasing Into Your Environmental Management System (EMS)

    EPA Pesticide Factsheets

    The goal of this report is to help Federal facilities integrate green purchasing into their EMS. The intended audience includes those tasked with implementing an EMS, reducing environmental impacts, meeting green purchasing requirements.

  17. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 2 2010-01-01 2010-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52...

  18. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 2 2014-01-01 2014-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52 ...

  19. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 2 2013-01-01 2013-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52 ...

  20. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 10 Energy 2 2012-01-01 2012-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52 ...

  1. 10 CFR Appendixes E-M to Part 52 - [Reserved

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 2 2011-01-01 2011-01-01 false E Appendixes E-M to Part 52 Energy NUCLEAR REGULATORY COMMISSION (CONTINUED) LICENSES, CERTIFICATIONS, AND APPROVALS FOR NUCLEAR POWER PLANTS Appendixes E-M to Part 52 ...

  2. Risk Communication Within the EM Program

    SciTech Connect

    Edelson, M.

    2003-02-26

    The U.S. Department of Energy Environmental Management program (EM) conducts the most extensive environmental remediation effort in the world. The annual EM budgets have exceeded $6,000,000,000 for approximately ten years and EM has assumed responsibility for the cleanup of the largest DOE reservations (i.e., at Hanford, Washington, Aiken, South Carolina, and Idaho Falls, Idaho) as well as the facilities at Rocky Flats, Colorado and in Ohio. Each of these sites has areas of extensive radioactive and chemical contamination, numerous surplus facilities that require decontamination and removal, while some have special nuclear material that requires secure storage. The EM program has been criticized for being ineffective (1) and has been repeatedly reorganized to address perceived shortcomings. The most recent reorganization was announced in 2001 to become effective at the beginning of the 2003 Federal Fiscal Year (i.e., October 2002). It was preceded by a ''top to bottom'' review (TTBR) of the program (2) that identified several deficiencies that were to be corrected as a result of the reorganization. One prominent outcome of the TTBR was the identification of ''risk reduction'' as an organizing principle to prioritize the activities of the new EM program. The new program also sought to accelerate progress by identifying a set of critical activities at each site that could be accelerated and result in more rapid site closure, with attendant risk, cost, and schedule benefits. This paper investigates how the new emphasis on risk reduction in the EM program has been communicated to EM stakeholders and regulators. It focuses on the Rocky Flats Environmental Technology Site (RFETS) as a case study and finds that there is little evidence for a new emphasis on risk reduction in EM communications with RFETS stakeholders. Discussions between DOE and RFETS stakeholders often refer to ''risk,'' but the word serves as a placeholder for other concepts. Thus ''risk'' communication

  3. Structural Composites With Tuned EM Chirality

    DTIC Science & Technology

    2014-12-23

    AFRL-OSR-VA-TR-2015-0018 STRUCTURAL COMPOSITES WITH TUNED EM CHIRALITY Siavouche Nemat Nasser UNIVERSITY OF CALIFORNIA SAN DIEGO Final Report 12/23...REPORT Grant/Contract  Title:        STRUCTURAL  COMPOSITES  WITH  TUNED  EM   CHIRALITY     Grant  No.:  FA9550-­‐09-­‐1...structural   composites  with   tunable   chiral   elements   has   produced   some   impressive   results   in   the

  4. Lessons Learned: Integrating EMS and Environmental Compliance Auditing

    DTIC Science & Technology

    2009-05-06

    audit an EMS against the – ISO 14001 :2004 Standard – Installation’s own commitments and procedures established under their EMS (i.e. are they doing...what they said they would do?) – Army Policy • Findings are documented in a Word report (external audits) • ISO 14001 conformance is determined by...regulatory requirements • EMS Findings – Conformance: management system conforms to ISO 14001 standard – Major Nonconformance: EMS is missing element

  5. Do earthquakes generate EM signals?

    NASA Astrophysics Data System (ADS)

    Walter, Christina; Onacha, Stephen; Malin, Peter; Shalev, Eylon; Lucas, Alan

    2010-05-01

    study areas, large swarms of earthquakes were located very close to the electromagnetic coils. This abstract focuses on the data from the Wairakei area. Preliminary data analysis has been carried out by band pass filtering and removing of the harmonics of the 50 Hz power line frequency. The initial results clearly show that electromagnetic signals accompany the seismic P and S waves (coseismic signal). Further data analysis involves the extraction of the seismoelectric signal generated at the onset of the earthquake and at interfaces from the coseismic signal and other ‘noise' sources. This processing step exhibits a major challenge in seismoelectric data processing. Unlike in other studies we measured the EM field and the seismic field at one location. Therefore the seismoelectric wave travelling at the speed of light cannot be determined as easily in the arrival times as when an array of coils is used. This makes the determination of the origin time much more difficult. Hence other processing techniques need to be explored.

  6. EMS response to an airliner crash.

    PubMed

    Dasgupta, Shuvra; French, Simone; Williams-Johnson, Jean; Hutson, Rhonda; Hart, Nicole; Wong, Mark; Williams, Eric; Espinosa, Kurdell; Maycock, Celeste; Edwards, Romayne; McCartney, Trevor; Cawich, Shamir; Crandon, Ivor

    2012-06-01

    This report of an aircraft crash at a major airport in Kingston, Jamaica examines the response of the local Emergency Medical Services (EMS). Factors that impacted the response are discussed, and the need for more disaster simulation exercises is highlighted. The objective of this case report was to document the response of EMS personnel to the crash of American Airlines Flight 331, and to utilize the information to examine and improve the present protocol. While multiple errors can occur during a mass-casualty event, these can be reduced by frequent simulation exercises during which various personnel practice and learn designated roles. Efficient triage, proper communication, and knowledge of the roles are important in ensuring the best possible outcome. While the triage system and response of the EMS personnel were effective for this magnitude of catastrophe, more work is needed in order to meet predetermined standards. Ways in which this can be overcome include: (1) hosting more disaster simulation exercises; (2) encouraging more involvement with first responders; and (3) strengthening the links in the local EMS system. Vigorous public education must be instituted and maintained.

  7. Navigating 3D electron microscopy maps with EM-SURFER.

    PubMed

    Esquivel-Rodríguez, Juan; Xiong, Yi; Han, Xusi; Guang, Shuomeng; Christoffer, Charles; Kihara, Daisuke

    2015-05-30

    The Electron Microscopy DataBank (EMDB) is growing rapidly, accumulating biological structural data obtained mainly by electron microscopy and tomography, which are emerging techniques for determining large biomolecular complex and subcellular structures. Together with the Protein Data Bank (PDB), EMDB is becoming a fundamental resource of the tertiary structures of biological macromolecules. To take full advantage of this indispensable resource, the ability to search the database by structural similarity is essential. However, unlike high-resolution structures stored in PDB, methods for comparing low-resolution electron microscopy (EM) density maps in EMDB are not well established. We developed a computational method for efficiently searching low-resolution EM maps. The method uses a compact fingerprint representation of EM maps based on the 3D Zernike descriptor, which is derived from a mathematical series expansion for EM maps that are considered as 3D functions. The method is implemented in a web server named EM-SURFER, which allows users to search against the entire EMDB in real-time. EM-SURFER compares the global shapes of EM maps. Examples of search results from different types of query structures are discussed. We developed EM-SURFER, which retrieves structurally relevant matches for query EM maps from EMDB within seconds. The unique capability of EM-SURFER to detect 3D shape similarity of low-resolution EM maps should prove invaluable in structural biology.

  8. The association between EMS workplace safety culture and safety outcomes.

    PubMed

    Weaver, Matthew D; Wang, Henry E; Fairbanks, Rollin J; Patterson, Daniel

    2012-01-01

    Prior studies have highlighted wide variation in emergency medical services (EMS) workplace safety culture across agencies. To determine the association between EMS workplace safety culture scores and patient or provider safety outcomes. We administered a cross-sectional survey to EMS workers affiliated with a convenience sample of agencies. We recruited these agencies from a national EMS management organization. We used the EMS Safety Attitudes Questionnaire (EMS-SAQ) to measure workplace safety culture and the EMS Safety Inventory (EMS-SI), a tool developed to capture self-reported safety outcomes from EMS workers. The EMS-SAQ provides reliable and valid measures of six domains: safety climate, teamwork climate, perceptions of management, working conditions, stress recognition, and job satisfaction. A panel of medical directors, emergency medical technicians and paramedics, and occupational epidemiologists developed the EMS-SI to measure self-reported injury, medical errors and adverse events, and safety-compromising behaviors. We used hierarchical linear models to evaluate the association between EMS-SAQ scores and EMS-SI safety outcome measures. Sixteen percent of all respondents reported experiencing an injury in the past three months, four of every 10 respondents reported an error or adverse event (AE), and 89% reported safety-compromising behaviors. Respondents reporting injury scored lower on five of the six domains of safety culture. Respondents reporting an error or AE scored lower for four of the six domains, while respondents reporting safety-compromising behavior had lower safety culture scores for five of the six domains. Individual EMS worker perceptions of workplace safety culture are associated with composite measures of patient and provider safety outcomes. This study is preliminary evidence of the association between safety culture and patient or provider safety outcomes.

  9. The association between EMS workplace safety culture and safety outcomes

    PubMed Central

    Weaver, Matthew D.; Wang, Henry E.; Fairbanks, Rollin J.; Patterson, Daniel

    2012-01-01

    Objective Prior studies have highlighted wide variation in EMS workplace safety culture across agencies. We sought to determine the association between EMS workplace safety culture scores and patient or provider safety outcomes. Methods We administered a cross-sectional survey to EMS workers affiliated with a convenience sample of agencies. We recruited these agencies from a national EMS management organization. We used the EMS Safety Attitudes Questionnaire (EMS-SAQ) to measure workplace safety culture and the EMS Safety Inventory (EMS-SI), a tool developed to capture self-reported safety outcomes from EMS workers. The EMS-SAQ provides reliable and valid measures of six domains: safety climate, teamwork climate, perceptions of management, perceptions of working conditions, stress recognition, and job satisfaction. A panel of medical directors, paramedics, and occupational epidemiologists developed the EMS-SI to measure self-reported injury, medical errors and adverse events, and safety-compromising behaviors. We used hierarchical linear models to evaluate the association between EMS-SAQ scores and EMS-SI safety outcome measures. Results Sixteen percent of all respondents reported experiencing an injury in the past 3 months, four of every 10 respondents reported an error or adverse event (AE), and 90% reported safety-compromising behaviors. Respondents reporting injury scored lower on 5 of the 6 domains of safety culture. Respondents reporting an error or AE scored lower for 4 of the 6 domains, while respondents reporting safety-compromising behavior had lower safety culture scores for 5 of 6 domains. Conclusions Individual EMS worker perceptions of workplace safety culture are associated with composite measures of patient and provider safety outcomes. This study is preliminary evidence of the association between safety culture and patient or provider safety outcomes. PMID:21950463

  10. Accuracy of EMS Trauma Transport Destination Plans in North Carolina.

    PubMed

    Moss, Chailee; Cowden, Christopher S; Atterton, Laurie Meyer; Arasaratnam, Meredith H; Fernandez, Antonio R; Evarts, Jeff S; Barrier, Brian; Lerner, E Brooke; Mann, N Clay; Lohmeier, Chad; Shofer, Frances S; Brice, Jane H

    2015-01-01

    Abstract Objective. Planning for time-sensitive injury may allow emergency medical services (EMS) systems to more accurately triage patients meeting accepted criteria to facilities most capable of providing life-saving treatment. In 2010, North Carolina (NC) implemented statewide Trauma Triage and Destination Plans (TTDPs) in all 100 of North Carolina's county-defined EMS systems. Each system was responsible for identifying the specific destination hospitals with appropriate resources to treat trauma patients. We sought to characterize the accuracy of their hospital designations. Methods. In this cross-sectional study, we collected TTDPs for each county-defined EMS system, including their assigned hospital capabilities (i.e., trauma center or community hospital). We conducted a survey with each EMS system to determine how their TTDP was constructed and maintained, as well as with each TTDP-designated hospital to verify their capabilities. We determined the accuracy of the EMS assigned hospital designations by comparing them to the hospital's reported capabilities. Results. The 100 NC EMS systems provided 380 designations for 112 hospitals. TTDPs were created by EMS administrators and medical directors, with only 55% of EMS systems engaging a hospital representative in the plan creation. Compared to the actual hospital capabilities, 97% of the EMS TTDP designations were correct. Twelve hospital designations were incorrect and the majority (10) overestimated hospital capabilities. Of the 100 EMS systems, 7 misclassified hospitals in their TTDP. EMS systems that did not verify their local hospitals' capabilities during TTDP development were more likely to incorrectly categorize a hospital's capabilities (p = 0.001). Conclusions. A small number of EMS systems misclassified hospitals in their TTDP, but most plans accurately reflected hospital capabilities. Misclassification occurred more often in systems that did not consult local hospitals prior to developing their

  11. What Do They Want from Us? A Survey of EM Program Directors on EM Application Criteria

    PubMed Central

    King, Kevin; Kass, Dara

    2017-01-01

    Introduction Although a relatively young specialty, emergency medicine (EM) is popular among medical students and is one of the most competitive large specialties. Consequently, students increasingly seek more opportunity to differentiate themselves from their colleagues by pursuing more clerkships at the cost of taking out additional loans: this despite the fact that those who match in EM typically do so in their top three choices. We sought to ascertain what factors EM program directors seek in their typical candidate. Methods We recruited EM program directors via the Council of Emergency Medicine Residency Directors email listserv to participate in an anonymous survey regarding the United States Medical Licensing Examination (USMLE), the number of standardized letters of evaluation (SLOE), and the number of EM rotations during the fourth year. Results 135 respondents completed the anonymous survey: 59% of respondents stated their program did not have a minimum USMLE Step 1 score, but 39% reported a minimum score of 210 or higher; 95% of programs do not require Step 2 to grant an interview, but 46% require it to place the student on the rank list; 80% require only one EM rotation to grant an interview and none require more than two; 95% of programs will accept two SLOEs for both application and rank list placement. Conclusion For the typical EM applicant, there is likely little benefit to performing more than two rotations and obtaining more than two SLOEs. Students can defer USMLE Step 2 but must complete it by the time rank lists are due. Our study was limited by the anonymity of the survey, and comments by the respondents revealed the questions did not account for some nuances programs apply to their application review process. PMID:28116023

  12. The E-MS Algorithm: Model Selection with Incomplete Data.

    PubMed

    Jiang, Jiming; Nguyen, Thuan; Rao, J Sunil

    2015-04-04

    We propose a procedure associated with the idea of the E-M algorithm for model selection in the presence of missing data. The idea extends the concept of parameters to include both the model and the parameters under the model, and thus allows the model to be part of the E-M iterations. We develop the procedure, known as the E-MS algorithm, under the assumption that the class of candidate models is finite. Some special cases of the procedure are considered, including E-MS with the generalized information criteria (GIC), and E-MS with the adaptive fence (AF; Jiang et al. 2008). We prove numerical convergence of the E-MS algorithm as well as consistency in model selection of the limiting model of the E-MS convergence, for E-MS with GIC and E-MS with AF. We study the impact on model selection of different missing data mechanisms. Furthermore, we carry out extensive simulation studies on the finite-sample performance of the E-MS with comparisons to other procedures. The methodology is also illustrated on a real data analysis involving QTL mapping for an agricultural study on barley grains.

  13. The E-MS Algorithm: Model Selection with Incomplete Data

    PubMed Central

    Jiang, Jiming; Nguyen, Thuan; Rao, J. Sunil

    2014-01-01

    We propose a procedure associated with the idea of the E-M algorithm for model selection in the presence of missing data. The idea extends the concept of parameters to include both the model and the parameters under the model, and thus allows the model to be part of the E-M iterations. We develop the procedure, known as the E-MS algorithm, under the assumption that the class of candidate models is finite. Some special cases of the procedure are considered, including E-MS with the generalized information criteria (GIC), and E-MS with the adaptive fence (AF; Jiang et al. 2008). We prove numerical convergence of the E-MS algorithm as well as consistency in model selection of the limiting model of the E-MS convergence, for E-MS with GIC and E-MS with AF. We study the impact on model selection of different missing data mechanisms. Furthermore, we carry out extensive simulation studies on the finite-sample performance of the E-MS with comparisons to other procedures. The methodology is also illustrated on a real data analysis involving QTL mapping for an agricultural study on barley grains. PMID:26783375

  14. Applying Recursive EM to Scene Segmentation

    NASA Astrophysics Data System (ADS)

    Bachmann, Alexander

    In this paper a novel approach for the interdependent task of multiple object tracking and scene segmentation is presented. The method partitions a stereo image sequence of a dynamic 3-dimensional (3D) scene into its most prominent moving groups with similar 3D motion. The unknown set of motion parameters is recursively estimated using an iterated extended Kalman filter (IEKF) which will be derived from the expectation-maximization (EM) algorithm. The EM formulation is used to incorporate a probabilistic data association measure into the tracking process. In a subsequent segregation step, each image point is assigned to the object hypothesis with maximum a posteriori (MAP) probability. Within the association process, which is implemented as labeling problem, a Markov Random Field (MRF) is used to express our expectations on spatial continuity of objects.

  15. Intense EM filamentation in relativistic hot plasmas

    NASA Astrophysics Data System (ADS)

    Hu, Qiang-Lin; Chen, Zhong-Ping; Mahajan, Swadesh M.

    2017-03-01

    Through 2D particle-in-cell (PIC) simulations, we demonstrate that the nature of filamentation of a high intensity electromagnetic (EM) pulse propagating in an underdense plasma, is profoundly affected at relativistically high temperatures. The "relativistic" filaments are sharper, are dramatically extended (along the direction of propagation), and live much longer than their lower temperature counterparts. The thermally boosted electron inertia is invoked to understand this very interesting and powerful phenomenon.

  16. Cryo-EM: Spinning the Micelles Away.

    PubMed

    Singh, Satinder K; Sigworth, Fred J

    2015-09-01

    Structural characterization of integral membrane proteins (MPs) demands that the samples be pure, monodisperse, and stable. Detergents are required to extract MPs from the lipid bilayer in which they reside and to stabilize them for downstream biophysical analyses. Some of the best MP-stabilizing detergents pose problems for cryo-EM studies, but in this issue of Structure, Hauer et al. (2015) now offer a solution called GraDeR.

  17. Leukocyte Recognition Using EM-Algorithm

    NASA Astrophysics Data System (ADS)

    Colunga, Mario Chirinos; Siordia, Oscar Sánchez; Maybank, Stephen J.

    This document describes a method for classifying images of blood cells. Three different classes of cells are used: Band Neutrophils, Eosinophils and Lymphocytes. The image pattern is projected down to a lower dimensional sub space using PCA; the probability density function for each class is modeled with a Gaussian mixture using the EM-Algorithm. A new cell image is classified using the maximum a posteriori decision rule.

  18. TandEM: Titan and Enceladus mission

    NASA Astrophysics Data System (ADS)

    Coustenis, A.; Atreya, S. K.; Balint, T.; Brown, R. H.; Dougherty, M. K.; Ferri, F.; Fulchignoni, M.; Gautier, D.; Gowen, R. A.; Griffith, C. A.; Gurvits, L. I.; Jaumann, R.; Langevin, Y.; Leese, M. R.; Lunine, J. I.; McKay, C. P.; Moussas, X.; Müller-Wodarg, I.; Neubauer, F.; Owen, T. C.; Raulin, F.; Sittler, E. C.; Sohl, F.; Sotin, C.; Tobie, G.; Tokano, T.; Turtle, E. P.; Wahlund, J.-E.; Waite, J. H.; Baines, K. H.; Blamont, J.; Coates, A. J.; Dandouras, I.; Krimigis, T.; Lellouch, E.; Lorenz, R. D.; Morse, A.; Porco, C. C.; Hirtzig, M.; Saur, J.; Spilker, T.; Zarnecki, J. C.; Choi, E.; Achilleos, N.; Amils, R.; Annan, P.; Atkinson, D. H.; Bénilan, Y.; Bertucci, C.; Bézard, B.; Bjoraker, G. L.; Blanc, M.; Boireau, L.; Bouman, J.; Cabane, M.; Capria, M. T.; Chassefière, E.; Coll, P.; Combes, M.; Cooper, J. F.; Coradini, A.; Crary, F.; Cravens, T.; Daglis, I. A.; de Angelis, E.; de Bergh, C.; de Pater, I.; Dunford, C.; Durry, G.; Dutuit, O.; Fairbrother, D.; Flasar, F. M.; Fortes, A. D.; Frampton, R.; Fujimoto, M.; Galand, M.; Grasset, O.; Grott, M.; Haltigin, T.; Herique, A.; Hersant, F.; Hussmann, H.; Ip, W.; Johnson, R.; Kallio, E.; Kempf, S.; Knapmeyer, M.; Kofman, W.; Koop, R.; Kostiuk, T.; Krupp, N.; Küppers, M.; Lammer, H.; Lara, L.-M.; Lavvas, P.; Le Mouélic, S.; Lebonnois, S.; Ledvina, S.; Li, J.; Livengood, T. A.; Lopes, R. M.; Lopez-Moreno, J.-J.; Luz, D.; Mahaffy, P. R.; Mall, U.; Martinez-Frias, J.; Marty, B.; McCord, T.; Menor Salvan, C.; Milillo, A.; Mitchell, D. G.; Modolo, R.; Mousis, O.; Nakamura, M.; Neish, C. D.; Nixon, C. A.; Nna Mvondo, D.; Orton, G.; Paetzold, M.; Pitman, J.; Pogrebenko, S.; Pollard, W.; Prieto-Ballesteros, O.; Rannou, P.; Reh, K.; Richter, L.; Robb, F. T.; Rodrigo, R.; Rodriguez, S.; Romani, P.; Ruiz Bermejo, M.; Sarris, E. T.; Schenk, P.; Schmitt, B.; Schmitz, N.; Schulze-Makuch, D.; Schwingenschuh, K.; Selig, A.; Sicardy, B.; Soderblom, L.; Spilker, L. J.; Stam, D.; Steele, A.; Stephan, K.; Strobel, D. F.; Szego, K.; Szopa, C.; Thissen, R.; Tomasko, M. G.; Toublanc, D.; Vali, H.; Vardavas, I.; Vuitton, V.; West, R. A.; Yelle, R.; Young, E. F.

    2009-03-01

    TandEM was proposed as an L-class (large) mission in response to ESA’s Cosmic Vision 2015-2025 Call, and accepted for further studies, with the goal of exploring Titan and Enceladus. The mission concept is to perform in situ investigations of two worlds tied together by location and properties, whose remarkable natures have been partly revealed by the ongoing Cassini-Huygens mission. These bodies still hold mysteries requiring a complete exploration using a variety of vehicles and instruments. TandEM is an ambitious mission because its targets are two of the most exciting and challenging bodies in the Solar System. It is designed to build on but exceed the scientific and technological accomplishments of the Cassini-Huygens mission, exploring Titan and Enceladus in ways that are not currently possible (full close-up and in situ coverage over long periods of time). In the current mission architecture, TandEM proposes to deliver two medium-sized spacecraft to the Saturnian system. One spacecraft would be an orbiter with a large host of instruments which would perform several Enceladus flybys and deliver penetrators to its surface before going into a dedicated orbit around Titan alone, while the other spacecraft would carry the Titan in situ investigation components, i.e. a hot-air balloon (Montgolfière) and possibly several landing probes to be delivered through the atmosphere.

  19. TandEM: Titan and Enceladus mission

    USGS Publications Warehouse

    Coustenis, A.; Atreya, S.K.; Balint, T.; Brown, R.H.; Dougherty, M.K.; Ferri, F.; Fulchignoni, M.; Gautier, D.; Gowen, R.A.; Griffith, C.A.; Gurvits, L.I.; Jaumann, R.; Langevin, Y.; Leese, M.R.; Lunine, J.I.; McKay, C.P.; Moussas, X.; Muller-Wodarg, I.; Neubauer, F.; Owen, T.C.; Raulin, F.; Sittler, E.C.; Sohl, F.; Sotin, Christophe; Tobie, G.; Tokano, T.; Turtle, E.P.; Wahlund, J.-E.; Waite, J.H.; Baines, K.H.; Blamont, J.; Coates, A.J.; Dandouras, I.; Krimigis, T.; Lellouch, E.; Lorenz, R.D.; Morse, A.; Porco, C.C.; Hirtzig, M.; Saur, J.; Spilker, T.; Zarnecki, J.C.; Choi, E.; Achilleos, N.; Amils, R.; Annan, P.; Atkinson, D.H.; Benilan, Y.; Bertucci, C.; Bezard, B.; Bjoraker, G.L.; Blanc, M.; Boireau, L.; Bouman, J.; Cabane, M.; Capria, M.T.; Chassefiere, E.; Coll, P.; Combes, M.; Cooper, J.F.; Coradini, A.; Crary, F.; Cravens, T.; Daglis, I.A.; de Angelis, E.; De Bergh, C.; de Pater, I.; Dunford, C.; Durry, G.; Dutuit, O.; Fairbrother, D.; Flasar, F.M.; Fortes, A.D.; Frampton, R.; Fujimoto, M.; Galand, M.; Grasset, O.; Grott, M.; Haltigin, T.; Herique, A.; Hersant, F.; Hussmann, H.; Ip, W.; Johnson, R.; Kallio, E.; Kempf, S.; Knapmeyer, M.; Kofman, W.; Koop, R.; Kostiuk, T.; Krupp, N.; Kuppers, M.; Lammer, H.; Lara, L.-M.; Lavvas, P.; Le, Mouelic S.; Lebonnois, S.; Ledvina, S.; Li, Ji; Livengood, T.A.; Lopes, R.M.; Lopez-Moreno, J. -J.; Luz, D.; Mahaffy, P.R.; Mall, U.; Martinez-Frias, J.; Marty, B.; McCord, T.; Salvan, C.M.; Milillo, A.; Mitchell, D.G.; Modolo, R.; Mousis, O.; Nakamura, M.; Neish, Catherine D.; Nixon, C.A.; Mvondo, D.N.; Orton, G.; Paetzold, M.; Pitman, J.; Pogrebenko, S.; Pollard, W.; Prieto-Ballesteros, O.; Rannou, P.; Reh, K.; Richter, L.; Robb, F.T.; Rodrigo, R.; Rodriguez, S.; Romani, P.; Bermejo, M.R.; Sarris, E.T.; Schenk, P.; Schmitt, B.; Schmitz, N.; Schulze-Makuch, D.; Schwingenschuh, K.; Selig, A.; Sicardy, B.; Soderblom, L.; Spilker, L.J.; Stam, D.; Steele, A.; Stephan, K.; Strobel, D.F.; Szego, K.; Szopa,

    2009-01-01

    TandEM was proposed as an L-class (large) mission in response to ESA's Cosmic Vision 2015-2025 Call, and accepted for further studies, with the goal of exploring Titan and Enceladus. The mission concept is to perform in situ investigations of two worlds tied together by location and properties, whose remarkable natures have been partly revealed by the ongoing Cassini-Huygens mission. These bodies still hold mysteries requiring a complete exploration using a variety of vehicles and instruments. TandEM is an ambitious mission because its targets are two of the most exciting and challenging bodies in the Solar System. It is designed to build on but exceed the scientific and technological accomplishments of the Cassini-Huygens mission, exploring Titan and Enceladus in ways that are not currently possible (full close-up and in situ coverage over long periods of time). In the current mission architecture, TandEM proposes to deliver two medium-sized spacecraft to the Saturnian system. One spacecraft would be an orbiter with a large host of instruments which would perform several Enceladus flybys and deliver penetrators to its surface before going into a dedicated orbit around Titan alone, while the other spacecraft would carry the Titan in situ investigation components, i.e. a hot-air balloon (Montgolfi??re) and possibly several landing probes to be delivered through the atmosphere. ?? Springer Science + Business Media B.V. 2008.

  20. The US DOE EM international program

    SciTech Connect

    Elmetti, Rosa R.; Han, Ana M.; Roach, Jay A.

    2013-07-01

    The U.S. Department of Energy (DOE) Office of Environmental Management (EM) conducts international collaboration activities in support of U.S. policies and objectives regarding the accelerated risk reduction and remediation of environmental legacy of the nations' nuclear weapons program and government sponsored nuclear energy research. The EM International Program supported out of the EM Office of the Associate Principal Deputy Assistant Secretary pursues collaborations with foreign government organizations, educational institutions and private industry to assist in identifying technologies and promote international collaborations that leverage resources and link international experience and expertise. In fiscal year (FY) 2012, the International Program awarded eight international collaborative projects for work scope spanning waste processing, groundwater and soil remediation, deactivation and decommissioning (D and D) and nuclear materials disposition initiatives to seven foreign organizations. Additionally, the International Program's scope and collaboration opportunities were expanded to include technical as well as non-technical areas. This paper will present an overview of the on-going tasks awarded in FY 2012 and an update of upcoming international activities and opportunities for expansion into the remainder of FY 2013 and beyond. (authors)

  1. Symptoms and reason for a medical visit in lung cancer patients.

    PubMed

    Gonzalez-Barcala, Francisco Javier; Falagan, José Antonio; Garcia-Prim, Jose Maria; Valdes, Luis; Carreira, Jose Martin; Pose, Antonio; Canive, Juan Carlos; Anton, Diana; Garcia-Sanz, Maria Teresa; Puga, Amalia; Temes, Enrique; Lopez-Lopes, Rafael

    2014-01-01

    Contexto: A Os doentes com o diagnóstico de cancro do pulmão estão habitualmente sintomáticos no momento do diagnóstico, sendo comum que o médico ou o doente não associem essa sintomatologia com a eventualidade de um tumor maligno. Objectivos: Este estudo teve como objectivo a análise dos sintomas de doentes com cancro do pulmão e sua relação com as características pessoais ou com a doença oncológica. Material e Métodos: Foi levado a cabo um estudo retrospectivo englobando todos os doentes com o diagnóstico de cancro do pulmão na Região de Saúde de Pontevedra (Espanha) ao longo de um período de três anos. São analisados os sintomas de apresentação do doente, o motivo de consulta e a concordância entre ambos ou com quaisquer factores correlacionados. Resultados: Foram incluídos no estudo 358 doentes, com uma média etária de 68,7 anos, sendo 87% dos doentes do sexo masculino. Os sintomas iniciais mais comuns foram sintomas constitucionais em 30,4% dos casos, tosse em 20,9% e dor torácica, descrita por 12% dos doentes. O motivo de consulta mais frequente foi dispneia em 22,1% dos doentes, um achado acidental em 15,4% dos doentes e hemoptise em 12,8%. Observou-se uma associação moderada (coeficiente de correlação = 0,495) entre os sintomas iniciais e o motivo de consulta. Conclusões: Uma elevada percentagem de doentes com um diagnóstico de cancro do pulmão apresentou sintomas associados com o tumor no momento do diagnóstico, mesmo num estadio inicial da doença.

  2. 2. HI PAR (ACQUISITION RADAR) TOWER AND ENLISTED MEN (EM) ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    2. HI PAR (ACQUISITION RADAR) TOWER AND ENLISTED MEN (EM) BARRACKS WITH RADAR ATTACHED. - Nike Hercules Missile Battery Summit Site, Battery Control Administration & Barracks Building, Anchorage, Anchorage, AK

  3. The State of Emergency Medical Services (EMS) Systems in Africa.

    PubMed

    Mould-Millman, Nee-Kofi; Dixon, Julia M; Sefa, Nana; Yancey, Arthur; Hollong, Bonaventure G; Hagahmed, Mohamed; Ginde, Adit A; Wallis, Lee A

    2017-02-23

    Introduction Little is known about the existence, distribution, and characteristics of Emergency Medical Services (EMS) systems in Africa, or the corresponding epidemiology of prehospital illness and injury.

  4. [Hypofractionation in locally advanced breast cancer: "flash" scheme].

    PubMed

    Padilha, Marisa; Gonçalves, Sara; Fardilha, Carlos; Melo, Gilberto; Miranda, Cristina; Alves, Paula

    2013-01-01

    Introdução: O carcinoma da mama é uma das principais causas de morte no nosso país. No Serviço de Radioterapia do Instituto Português de Oncologia de Coimbra de Coimbra utilizamos, desde há mais de 30 anos, um esquema de hipofraccionamento de radioterapia, denominado “Flash”, como opção terapêutica em doentes idosos ou com baixo Performance Status, portadores de carcinoma da mama localmente avançado ou com estádios IIb ou IV, com intenção neoadjuvante ou paliativa. Objectivos: Avaliar a resposta ao tratamento, nomeadamente sobrevivência global aos três anos, resposta local e toxicidades aguda e crónica, no grupo de doentes seleccionados submetidos a esquema de hipofraccionamento, em estudo retrospectivo. Metodologia: Entre Janeiro de 2006 e Dezembro de 2008, um total de 83 doentes com diagnóstico de Carcinoma da Mama Localmente Avançado ou com estádios IIb ou IV, foi submetido a “Flash” mamário. A dose de radioterapia prescrita foi de 13Gy / 2Fr / 3 dias (em 23 doentes - 27,7%) e 26Gy / 4Fr / 2,5 semanas (em 60 doentes - 72,3%), com fotões de 4 MV, sobre a mama afectada. Foi avaliada sobrevivência global segundo o método de Kaplan-Meier. A análise estatística foi efectuada através da aplicação SPSS, versão 17.0 e os testes estatísticos foram avaliados ao nível de significância de 5%. Resultados: 80 doentes (96,4%) que efectuaram “Flash” mamário eram do género feminino, com idades compreendidas entre os 59 e os 93 anos (idade média 80,72 + 5,87 anos) e Performance Status (Karnosfsky: 0 - 100) entre 90 e 50%. Em 72 doentes (86,7%) o diagnóstico histológico foi Carcinoma Ductal Invasivo. A cirurgia após a realização do “Flash” Mamário foi realizada em 44 doentes (53%) após evidência de resposta local à radioterapia, sendo a Mastectomia Radical Modificada a técnica cirúrgica mais frequente. Efectuou-se o diagnóstico de metastização óssea em 10 doentes (12%), sendo que a taxa de sobrevivência global foi

  5. Recruitment and retention in rural and urban EMS: results from a national survey of local EMS directors.

    PubMed

    Freeman, Victoria A; Slifkin, Rebecca T; Patterson, P Daniel

    2009-01-01

    Maintaining an adequate staff is a challenge for rural emergency medical services (EMS) providers. This national survey of local EMS directors finds that rural EMS are more likely to be freestanding, that is, not affiliated with other public services, to employ only emergency medical technician-basics (EMT-Bs), and to be all volunteer. Rural EMS directors are more likely than urban ones to report that they are not currently fully staffed. The most common barriers to recruitment of EMTs in both urban and rural areas include unwillingness of community members to volunteer and lack of certified EMTs in the area. In rural areas, barriers to EMT training were noted more often than in urban areas as was the lack of employer support for employee volunteers. Similar rural training barriers affected retention of staff. Rural respondents reported that they lose staff to burnout and to difficulty in meeting continuing education requirements. Among rural respondents, those who direct all-volunteer EMS were the most likely to report recruitment and retention problems. The results suggest areas for further study including how volunteer EMS agencies can transition to paid agencies, how to bring EMS education to rural areas, and how EMS can work with other agencies to ensure EMS viability.

  6. Evaluation of functional recovery by motor functional independence measure test of elderly after hip fracture in Serbia.

    PubMed

    Radosavljevic, Natasa; Nikolic, Dejan; Lazovic, Milica; Radosavljevic, Zoran; Jeremic, Aleksandar

    2014-01-01

    Introdução: o objectivo do estudo é a avaliação do nível de independência funcional através da aplicação da escala de Medida da Independência Funcional (Functional Independence Measure [FIM]) em doentes com idade superior a 65 anos, após fractura da anca.Material e Métodos: Foram estudados 203 doentes após fractura da anca, aplicando a escala MIF em 3 momentos: admissão do doente no hospital (Período 1), no momento da alta (Período 2) e 3 meses após a alta (Período 3); os doentes foram englobados em 3 grupos etários: Grupo65-74 , Grupo75-84 e Grupo+85 e em dois grupos, consoante o Índice de Gravidade (IG): grupo 0-1,99 (IG1) e grupo ≥ 2 (IG2).Resultados: No grupo de doentes com idêntico IG, observou-se um aumento dos valores da MIF no Período 2 e 3 em ambos osgéneros e nas primeiras duas classes etárias, ao passo que em doentes acima dos 85 anos, com IG mais elevado, observámos uma variação não significativa dos valores da MIF entre o momento da alta hospitalar e 3 meses após a alta.Discussão: A melhoria mais significativa da MIF foi obtida em doentes do sexo feminino no primeiro e terceiro grupos etários e com IG mais elevado.Conclusão: O género não constitui um factor preditivo significativo da recuperação da independência funcional medida através da aplicação da escala MIF em doentes com fractura da anca, embora a MIF no momento da admissão seja um bom indicador de recuperação funcional em doentes do sexo feminino em certos grupos etários (primeiro e terceiro grupos etários).

  7. Communication - An Effective Tool for Implementing ISO 14001/EMS

    SciTech Connect

    Rachel Damewood; Bowen Huntsman

    2004-04-01

    The Idaho National Engineering and Environmental Laboratory (INEEL) received ISO 14001/EMS certification in June 2002. Communication played an effective role in implementing ISO 14001/EMS at the INEEL. This paper describes communication strategies used during the implementation and certification processes. The INEEL achieved Integrated Safety Management System (ISMS) and Voluntary Protection Program (VPP) Star status in 2001. ISMS implemented a formal process to plan and execute work. VPP facilitated worker involvement by establishing geographic units at various facilities with employee points of contact and management champions. The INEEL Environmental Management System (EMS) was developed to integrate the environmental functional area into its ISMS and VPP. Since the core functions of ISMS, VPP, and EMS are interchangeable, they were easy to integrate. Communication is essential to successfully implement an EMS. (According to ISO 14001 requirements, communication interacts with 12 other elements of the requirements.) We developed communication strategies that integrated ISMS, VPP, and EMS. For example, the ISMS, VPP, and EMS Web sites communicated messages to the work force, such as “VPP emphasizes the people side of doing business, ISMS emphasizes the system side of doing business, and EMS emphasizes the systems to protect the environment; but they all define work, identify and analyze hazards, and mitigate the hazards.” As a result of this integration, the work force supported and implemented the EMS. In addition, the INEEL established a cross-functional communication team to assist with implementing the EMS. The team included members from the Training and Communication organizations, VPP office, Pollution Prevention, Employee and Media Relations, a union representative, facility environmental support, and EMS staff. This crossfunctional team used various communication strategies to promote our EMS to all organization levels and successfully implemented EMS

  8. Improving Emergency Medical Services (EMS) in the United States Through Improved and Centralized Federal Coordination

    DTIC Science & Technology

    2011-03-01

    other emergencies. 4 • Public Awareness of the EMS Discipline. The public at large is not aware of the EMS structure in its community or at a higher...regard for its imperative role in community service. For this reason, placing EMS under the USFA at the federal level would cause EMS to still be...service to include EMS is appropriate, and in some cases, it is not. Not all communities have a dedicated fire service, which can make EMS provision

  9. EM threat analysis for wireless systems.

    SciTech Connect

    Burkholder, R. J. (Ohio State University Electroscience Laboratory); Mariano, Robert J.; Schniter, P. (Ohio State University Electroscience Laboratory); Gupta, I. J. (Ohio State University Electroscience Laboratory)

    2006-06-01

    Modern digital radio systems are complex and must be carefully designed, especially when expected to operate in harsh propagation environments. The ability to accurately predict the effects of propagation on wireless radio performance could lead to more efficient radio designs as well as the ability to perform vulnerability analyses before and after system deployment. In this report, the authors--experts in electromagnetic (EM) modeling and wireless communication theory--describe the construction of a simulation environment that is capable of quantifying the effects of wireless propagation on the performance of digital communication.

  10. Association between EMS Question Bank Completion and Passing Rates on the EMS Certification Examination.

    PubMed

    Clemency, Brian; Martin-Gill, Christian; Rall, Nicole; May, Paul; Lubin, Jeffrey; Cooley, Craig; Van Dillen, Christine; Silvestri, Salvatore; Portela, Roberto; Cooney, Derek; Knutsen, Christian; March, Juan

    2017-01-01

    A board review question bank was created to assist candidates in their preparation for the 2015 EMS certification examination. We aimed to describe the development of this question bank and evaluate its successes in preparing candidates to obtain EMS subspecialty board certification. An online question bank was developed by 13 subject matter experts who participated as item writers, representing eight different EMS fellowship programs. The online question bank consisted of four practice tests, with each of the tests comprised of 100 questions. The number of candidates who participated in and completed the question bank was calculated. The passing rate among candidates who completed the question bank was calculated and compared to the publicly reported statistics for all candidates. The relationship between candidates' performance on the question bank and subspecialty exam pass rates was determined. A total of 252 candidates took at least one practice test and, of those, 225 candidates completed all four 100-question practice tests. The pass rate on the 2015 EMS certification exam was 79% (95%CI 74-85%) among candidates who completed the question bank, which is 12% higher than the overall pass rate (p = 0.003). Candidates' performance on the question bank was positively associated with overall success on the exam (X(2) = 75.8, p < 0.0001). Achieving a score of ≥ 70% on the question bank was associated with a higher likelihood of passing the exam (OR = 17.8; 95% CI: 8.0-39.6). Completing the question bank program was associated with improved pass rates on the EMS certification exam. Strong performance on the question bank correlated with success on the exam.

  11. Light curve analysis of southern eclipsing binary EM Car

    NASA Astrophysics Data System (ADS)

    Ćiçek, C.; Bulut, I.; Bulut, A.

    2017-02-01

    In this study, ASAS light curve of the eclipsing binary EM Car (Sp = O8V, P = 3.4 days) has been analyzed using the Wilson-Devinney method. The light curve analyses have found that EM Car is a detached eclipsing binary system with small eccentric orbit

  12. Genetic-based EM algorithm for learning Gaussian mixture models.

    PubMed

    Pernkopf, Franz; Bouchaffra, Djamel

    2005-08-01

    We propose a genetic-based expectation-maximization (GA-EM) algorithm for learning Gaussian mixture models from multivariate data. This algorithm is capable of selecting the number of components of the model using the minimum description length (MDL) criterion. Our approach benefits from the properties of Genetic algorithms (GA) and the EM algorithm by combination of both into a single procedure. The population-based stochastic search of the GA explores the search space more thoroughly than the EM method. Therefore, our algorithm enables escaping from local optimal solutions since the algorithm becomes less sensitive to its initialization. The GA-EM algorithm is elitist which maintains the monotonic convergence property of the EM algorithm. The experiments on simulated and real data show that the GA-EM outperforms the EM method since: 1) We have obtained a better MDL score while using exactly the same termination condition for both algorithms. 2) Our approach identifies the number of components which were used to generate the underlying data more often than the EM algorithm.

  13. CryoEM at IUCrJ: a new era

    PubMed Central

    Subramaniam, Sriram; Kühlbrandt, Werner; Henderson, Richard

    2016-01-01

    In this overview, we briefly outline recent advances in electron cryomicroscopy (cryoEM) and explain why the journal IUCrJ, published by the International Union of Crystallography, could provide a natural home for publications covering many present and future developments in the cryoEM field. PMID:26870375

  14. 7 CFR 759.6 - EM to be made available.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... natural disaster has occurred in a county, resulting in severe physical losses. If the FSA Administrator determines that such a natural disaster has occurred, then EM can be made available to eligible farmers for... AGRICULTURE SPECIAL PROGRAMS DISASTER DESIGNATIONS AND NOTIFICATIONS § 759.6 EM to be made available. (a)...

  15. 7 CFR 759.6 - EM to be made available.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... natural disaster has occurred in a county, resulting in severe physical losses. If the FSA Administrator determines that such a natural disaster has occurred, then EM can be made available to eligible farmers for... AGRICULTURE SPECIAL PROGRAMS DISASTER DESIGNATIONS AND NOTIFICATIONS § 759.6 EM to be made available. (a)...

  16. [PET/CT with 18F-Fluorocholine in Patients with Prostatic Cancer in Biochemical Recurrence].

    PubMed

    Lapa, Paula; Silva, Rodolfo; Saraiva, Tiago; Figueiredo, Arnaldo; Ferreira, Rui; Costa, Gracinda; Lima, João Pedroso

    2016-03-01

    Introdução: No carcinoma da próstata, é frequente, após terapêutica com intuito curativo, ocorrer recidiva bioquímica. O objectivo deste trabalho foi avaliar o impacto da PET/CT com fluorocolina-F18 no restadiamento e orientação destes doentes e analisar a influência, da estratificação de risco, dos valores do PSA e da terapêutica de supressão hormonal, na sensibilidade da técnica. Material e Métodos: Análise retrospectiva de 107 doentes com carcinoma da próstata em recidiva bioquímica que realizaram PET/CT com fluorocolina-F18 no nosso hospital, entre dezembro de 2009 e maio de 2014. Resultados: A sensibilidade global foi de 63,2% sendo 80,0% quando PSA > 2 ng/mL. Foi possível identificar doença à distância em 28% dos doentes. A sensibilidade aumentou de 40,0% em doentes de risco baixo e intermédio para 55,2% em doentes de alto risco. Sem terapêutica de supressão hormonal, a sensibilidade foi de 61,8% enquanto no grupo sob essa terapêutica, foi de 67,7%. Discussão: A PET/CT com fluorocolina-F18 forneceu informações relevantes, mesmo em doentes com baixos valores do PSA, contudo, com incremento significativo da sensibilidade nos doentes com PSA >2 ng/mL. A sensibilidade foi superior nos doentes de alto risco comparativamente com os de risco baixo e intermédio, contudo, sem uma diferença estatisticamente significativa. A terapêutica de supressão hormonal parece não influenciar a captação de Fluorocolina-F18 nos doentes resistentes à castração. Conclusões: Neste estudo, a PET/CT com fluorocolina-F18 apresentou bons resultados no restadiamento de doentes com carcinoma da próstata em recidiva bioquímica, distinguindo entre doença loco-regional e sistémica, informação com importantes consequências na definição da estratégia terapêutica.

  17. The Post-LBV Supernova 2001em

    NASA Astrophysics Data System (ADS)

    Van Dyk, Schuyler D.; Chornock, R.; Filippenko, A. V.; Foley, R. J.; Lewin, W. H. G.; Li, W.; Panagia, N.; Pooley, D.; Stockdale, C. J.; Weiler, K. W.

    2009-12-01

    The supernova (SN) 2001em in UGC 11794 was classified early as Type Ib/c, i.e., as one arising from a hydrogen-stripped star. As part of a radio survey with the Very Large Array of SNe Ib/c at late times (Stockdale et al. 2003, BAAS, 35, 1346), SN 2001em was detected as a highly luminous radio source ˜2 years after explosion. The SN was also subsequently discovered with Chandra to be a very luminous X-ray source. The properties of both the radio and X-ray emission are more characteristic of the Type II-narrow (IIn) SNe, where the SN shock is interacting with dense, massive circumstellar matter, resulting in bright radio synchrotron emission and thermal bremsstrahlung from the interaction region. In fact, SN 2001em has shown to have spectroscopically transformed to a SN IIn. The premise that this might indicate an off-axis gamma-ray burst has been presented (Granot & Ramirez-Ruiz 2004, ApJ, 609, L9) and later, rather convincingly, refuted (e.g., Schinzel et al. 2009, ApJ, 691, 1380). Chugai & Chevalier (2006, ApJ, 641, 1051) have interpreted the spectral transformation and radio/X-ray emission as the SN shock overtaking the detached hydrogen envelope of the progenitor star, which was shed in a superwind episode many years prior to explosion. Chevalier (2007, RMxAC, 30, 41) has further pointed to the required mass-loss rate in the event being equivalent to what would occur in the eruption of a luminous blue variable (LBV). The optical (ground-based and HST) and radio/X-ray data, together with mid-infrared Spitzer observations, tend to support this scenario of a very massive star that experienced a powerful LBV outburst prior to explosion. Such an event may not be unique, with possible parallels in the cases of SNe 2005bf and 2006jc, and may provide valuable new information about massive stellar evolution.

  18. Refinement of atomic models in high resolution EM reconstructions using Flex-EM and local assessment.

    PubMed

    Joseph, Agnel Praveen; Malhotra, Sony; Burnley, Tom; Wood, Chris; Clare, Daniel K; Winn, Martyn; Topf, Maya

    2016-05-01

    As the resolutions of Three Dimensional Electron Microscopic reconstructions of biological macromolecules are being improved, there is a need for better fitting and refinement methods at high resolutions and robust approaches for model assessment. Flex-EM/MODELLER has been used for flexible fitting of atomic models in intermediate-to-low resolution density maps of different biological systems. Here, we demonstrate the suitability of the method to successfully refine structures at higher resolutions (2.5-4.5Å) using both simulated and experimental data, including a newly processed map of Apo-GroEL. A hierarchical refinement protocol was adopted where the rigid body definitions are relaxed and atom displacement steps are reduced progressively at successive stages of refinement. For the assessment of local fit, we used the SMOC (segment-based Manders' overlap coefficient) score, while the model quality was checked using the Qmean score. Comparison of SMOC profiles at different stages of refinement helped in detecting regions that are poorly fitted. We also show how initial model errors can have significant impact on the goodness-of-fit. Finally, we discuss the implementation of Flex-EM in the CCP-EM software suite. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  19. Refinement of atomic models in high resolution EM reconstructions using Flex-EM and local assessment

    PubMed Central

    Joseph, Agnel Praveen; Malhotra, Sony; Burnley, Tom; Wood, Chris; Clare, Daniel K.; Winn, Martyn; Topf, Maya

    2016-01-01

    As the resolutions of Three Dimensional Electron Microscopic reconstructions of biological macromolecules are being improved, there is a need for better fitting and refinement methods at high resolutions and robust approaches for model assessment. Flex-EM/MODELLER has been used for flexible fitting of atomic models in intermediate-to-low resolution density maps of different biological systems. Here, we demonstrate the suitability of the method to successfully refine structures at higher resolutions (2.5–4.5 Å) using both simulated and experimental data, including a newly processed map of Apo-GroEL. A hierarchical refinement protocol was adopted where the rigid body definitions are relaxed and atom displacement steps are reduced progressively at successive stages of refinement. For the assessment of local fit, we used the SMOC (segment-based Manders’ overlap coefficient) score, while the model quality was checked using the Qmean score. Comparison of SMOC profiles at different stages of refinement helped in detecting regions that are poorly fitted. We also show how initial model errors can have significant impact on the goodness-of-fit. Finally, we discuss the implementation of Flex-EM in the CCP-EM software suite. PMID:26988127

  20. EMS incident management: emergency medical logistics.

    PubMed

    Maniscalco, P M; Christen, H T

    1999-01-01

    If you had to get x amount of supplies to point A or point B, or both, in 10 minutes, how would you do it? The answer lies in the following steps: 1. Develop a logistics plan. 2. Use emergency management as a partner agency for developing your logistics plan. 3. Implement a push logistics system by determining what supplies/medications and equipment are important. 4. Place mass casualty/disaster caches at key locations for rapid deployment. Have medication/fluid caches available at local hospitals. 5. Develop and implement command caches for key supervisors and managers. 6. Anticipate the logistics requirements of a terrorism/tactical violence event based on a community threat assessment. 7. Educate the public about preparing a BLS family disaster kit. 8. Test logistics capabilities at disaster exercises. 9. Budget for logistics needs. 10. Never underestimate the importance of logistics. When logistics support fails, the EMS system fails.

  1. A new preparedness policy for EMS logistics.

    PubMed

    Lee, Seokcheon

    2017-03-01

    Response time in emergency medical services (EMS) is defined as the interval for an ambulance to arrive the scene after receipt of a 911 call. When several ambulances are available upon the receipt of a new call, a decision of selecting an ambulance has to be made in an effort to reduce response time. Dispatching the closest unit available is commonly used in practice; however, recently the Preparedness policy was designed that is in a simplistic form yet being capable of securing a long-term efficiency. This research aims to improve the Preparedness policy, resolving several critical issues inherent in the current form of the policy. The new Preparedness policy incorporates a new metric of preparedness based on the notion of centrality and involves a tuning parameter, weight on preparedness, which has to be appropriately chosen according to operational scenario. Computational experiment shows that the new policy significantly improves the former policy robustly in various scenarios.

  2. Online EM with weight-based forgetting.

    PubMed

    Celaya, Enric; Agostini, Alejandro

    2015-05-01

    In the online version of the EM algorithm introduced by Sato and Ishii ( 2000 ), a time-dependent discount factor is introduced for forgetting the effect of the old estimated values obtained with an earlier, inaccurate estimator. In their approach, forgetting is uniformly applied to the estimators of each mixture component depending exclusively on time, irrespective of the weight attributed to each unit for the observed sample. This causes an excessive forgetting in the less frequently sampled regions. To address this problem, we propose a modification of the algorithm that involves a weight-dependent forgetting, different for each mixture component, in which old observations are forgotten according to the actual weight of the new samples used to replace older values. A comparison of the time-dependent versus the weight-dependent approach shows that the latter improves the accuracy of the approximation and exhibits much greater stability.

  3. DOE EM industry programs robotics development

    SciTech Connect

    Staubly, R.; Kothari, V.

    1997-12-01

    The Office of Science and Technology (OST) manages an aggressive program for RD&D, as well as testing and evaluation for the U.S. Department of Energy`s (DOE) Environmental Management (EM) organization. The goal is to develop new and improved environmental restoration and waste management technologies to clean up the inventory of the DOE weapons complex faster, safer, and cheaper than is possible with currently available technologies. OST has organized technology management activities along focus teams for each major problem area. There are currently five focus areas: decontamination and decommissioning, tanks, subsurface contaminants, mixed waste, and plutonium. In addition, OST is pursuing research and development (R&D) that cuts across these focus areas by having applications in two or more focus areas. Currently, there are three cross-cutting programs: the robotics technology development; characterization, monitoring, and sensor technologies; and efficient separations and processing.

  4. Epidemiology of Emergency Medical Services (EMS) Utilization in Four Indian Emergency Departments.

    PubMed

    Wijesekera, Olindi; Reed, Amanda; Chastain, Parker S; Biggs, Shauna; Clark, Elizabeth G; Kole, Tamorish; Chakrapani, Anoop T; Ashish, Nandy; Rajhans, Prasad; Breaud, Alan H; Jacquet, Gabrielle A

    2016-12-01

    Introduction Without a universal Emergency Medical Services (EMS) system in India, data on the epidemiology of patients who utilize EMS are limited. This retrospective chart review aimed to quantify and describe the burden of disease and patient demographics of patients who arrived by EMS to four Indian emergency departments (EDs) in order to inform a national EMS curriculum.

  5. Best Practices for Managing Large CryoEM Facilities

    PubMed Central

    Alewijnse, Bart; Ashton, Alun W.; Chambers, Melissa G.; Chen, Songye; Cheng, Anchi; Ebrahim, Mark; Eng, Edward; Hagen, Wim J. H.; Koster, Abraham J.; López, Claudia S.; Lukoyanova, Natalya; Ortega, Joaquin; Renault, Ludovic; Reyntjens, Steve; Rice, William J.; Scapin, Giovanna; Schrijver, Raymond; Siebert, Alistair; Stagg, Scott M.; Grum-Tokars, Valerie; Wright, Elizabeth R.; Wu, Shenping; Yu, Zhiheng; Zhou, Z. Hong; Carragher, Bridget; Potter, Clinton S.

    2017-01-01

    This paper provides an overview of the discussion and presentations from the Workshop on the Management of Large CryoEM Facilities held at the New York Structural Biology Center, New York, NY on February 6–7, 2017. A major objective of the workshop was to discuss best practices for managing cryoEM facilities. The discussions were largely focused on supporting single-particle methods for cryoEM and topics included: user access, assessing projects, workflow, sample handling, microscopy, data management and processing, and user training. PMID:28827185

  6. Environmental Education and Development Division (EM-522). Annual report, Fiscal year 1993

    SciTech Connect

    Not Available

    1993-12-31

    The Environmental Education and Development Division (EM-522) is one of three divisions within the Office of Technology Integration and Environmental Education and Development (EM-52) in Environmental Restoration and Waste Management`s (EM`s) Office of Technology Development (EM-50). The primary design criterion for EM-522 education activities is directly related to meeting EM`s goal of environmental compliance on an accelerated basis and cleanup of the 1989 inventory of inactive sites and facilities by the year 2019. Therefore, EM-522`s efforts are directed specifically toward stimulating knowledge and capabilities to achieve the goals of EM while contributing to DOE`s overall goal of increasing scientific, mathematical, and technical literacy and competency. This report discusses fiscal year 1993 activities.

  7. Transforming EMS Compliance at the Utah National Guard with an Automated Tool

    DTIC Science & Technology

    2009-05-01

    14001 & EMS 16 ISO 14001 EMS Requirements ISO14001 Requirement Condition WebEMIS™ 4.1 General Requirements None Uploads and stores EO 13423, EO...top level management with metrics and performance measures based on media WebEMIS vs. ISO 14001 & EMS 17 ISO 14001 EMS Requirements ISO14001 ...14001 EMS Requirements ISO14001 Requirement Condition WebEMIS 4.4 Implementation and Operation 4.4.1 Resources, Roles, Responsibility and Authority

  8. Recent developments in the CCP-EM software suite

    PubMed Central

    Burnley, Tom

    2017-01-01

    As part of its remit to provide computational support to the cryo-EM community, the Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) has produced a software framework which enables easy access to a range of programs and utilities. The resulting software suite incorporates contributions from different collaborators by encapsulating them in Python task wrappers, which are then made accessible via a user-friendly graphical user interface as well as a command-line interface suitable for scripting. The framework includes tools for project and data management. An overview of the design of the framework is given, together with a survey of the functionality at different levels. The current CCP-EM suite has particular strength in the building and refinement of atomic models into cryo-EM reconstructions, which is described in detail. PMID:28580908

  9. E.M. and Hadronic Shower Simulation with FLUKA

    SciTech Connect

    Battistoni, G.; Fasso, A.; Ferrari, A.; Ranft, J.; Rubbia, A.; Sala, P.R.; /INFN, Milan /SLAC /CERN /Siegen U. /Zurich, ETH

    2005-10-03

    A description of the main features of e.m. and hadronic shower simulation models used in the FLUKA code is summarized and some recent applications are discussed. The general status of the FLUKA project is also reported.

  10. Recent developments in the CCP-EM software suite.

    PubMed

    Burnley, Tom; Palmer, Colin M; Winn, Martyn

    2017-06-01

    As part of its remit to provide computational support to the cryo-EM community, the Collaborative Computational Project for Electron cryo-Microscopy (CCP-EM) has produced a software framework which enables easy access to a range of programs and utilities. The resulting software suite incorporates contributions from different collaborators by encapsulating them in Python task wrappers, which are then made accessible via a user-friendly graphical user interface as well as a command-line interface suitable for scripting. The framework includes tools for project and data management. An overview of the design of the framework is given, together with a survey of the functionality at different levels. The current CCP-EM suite has particular strength in the building and refinement of atomic models into cryo-EM reconstructions, which is described in detail.

  11. DOE-EM Science of Safety Robotics Challenge

    SciTech Connect

    Rimando, Rodrigo; Watts, Alex; Bobbitt, John; McLaughlin, Doug; Quigley, Morgan; Gladwell, Scott; McLoughlin, Mike; Kinnamon, Tony; Garcia, Joe; Ansari, Alex; Voyles, Richard; Chambers, David; Pryor, Mitch; Workman, Theresa; Mehling, Joshua; Browning, Kimberly; Deuel, Jake; Profitt, Bryan; Reibold, Marty

    2016-09-20

    During the week of August 22nd, 2016, over 150 technologists, stakeholders, and Department of Energy, Office of Environmental Management workers, met at DOE’s Portsmouth Gaseous Diffusion Plant in Ohio, for the EM Science of Safety Robotics Challenge.

  12. Near-atomic-resolution cryo-EM for molecular virology.

    PubMed

    Hryc, Corey F; Chen, Dong-Hua; Chiu, Wah

    2011-08-01

    Electron cryo-microscopy (cryo-EM) is a technique in structural biology that is widely used to solve the three-dimensional structures of macromolecular assemblies, close to their biological and solution conditions. Recent improvements in cryo-EM and single-particle reconstruction methodologies have led to the determination of several virus structures at near-atomic resolution (3.3 - 4.6 Å). These cryo-EM structures not only resolve the Cα backbones and side-chain densities of viral capsid proteins, but also suggest functional roles that the protein domains and some key amino acid residues play. This paper reviews the recent advances in near-atomic-resolution cryo-EM for probing the mechanisms of virus assembly and morphogenesis.

  13. EMS: A framework for data acquisition and analysis

    NASA Astrophysics Data System (ADS)

    Nogiec, J. M.; Sim, J.; Trombly-Freytag, K.; Walbridge, D.

    2001-08-01

    The Extensible Measurement System (EMS) is a universal Java framework for building data analysis and test systems. The objective of the EMS project is to replace a multitude of different existing systems with a single expandable system, capable of accommodating various test and analysis scenarios and varying algorithms. The EMS framework is based on component technology, graphical assembly of systems, introspection and flexibility to accommodate various data processing and data acquisition components. Core system components, common to many application domains, have been identified and designed together with the domain-specific components for the measurement of accelerator magnets. The EMS employs several modern technologies and the result is a highly portable, configurable, and potentially distributed system, with the capability of parallel signal data processing, parameterized test scripting, and run-time reconfiguration.

  14. EmPOWER Maryland - Leveraging relationships and experience

    EPA Pesticide Factsheets

    DHCD has gained a deep understanding of the needs of Maryland’s low-income residents and therefore was well-positioned to implement the low-income components of the broader EmPOWER Maryland energy efficiency initiative.

  15. Classification of ASASSN-17em/AT2017cts

    NASA Astrophysics Data System (ADS)

    Bersier, David

    2017-04-01

    We obtained a spectrum of the candidate supernova ASASSN-17em/AT2017cts (ATEL #10241), using the SPRAT spectrograph mounted on the robotic 2m Liverpool Telescope at the Roque de los Muchachos observatory (La Palma).

  16. Classification of ASASSN-17em/AT2017cts

    NASA Astrophysics Data System (ADS)

    Bersier, David

    2017-04-01

    We obtained a spectrum of the candidate supernova ASASSN-17em/AT2017cts (ATEL #10241), using the SPRAT spectrograph mounted on the robotic 2m Liverpool Telescope at the Roque de los Muchachos observatory (La Palma).

  17. Analysis of NSWC Ocean EM Observatory Test Data

    DTIC Science & Technology

    2016-09-01

    Analysis of NSWC Ocean EM Observatory test data: final report J. Bradley Nelson Date: September 2016 Contract #: NICOP - N62909-15...From - To) 09/01/2016 Technical Report #2 July 2015 - September 2016 4. TITLE AND SUBTITLE Sa. CONTRACT NUMBER Analysis of NSWC Ocean EM Observatory...distribution is unlimited 13. SUPPLEMENTARY NOTES 14. ABSTRACT NSWC-Carderock is leading a defense/academia group to instrument the South Florida Ocean

  18. NASA EM Followup of LIGO-Virgo Candidate Events

    NASA Technical Reports Server (NTRS)

    Blackburn, Lindy L.

    2011-01-01

    We present a strategy for a follow-up of LIGO-Virgo candidate events using offline survey data from several NASA high-energy photon instruments aboard RXTE, Swift, and Fermi. Time and sky-location information provided by the GW trigger allows for a targeted search for prompt and afterglow EM signals. In doing so, we expect to be sensitive to signals which are too weak to be publicly reported as astrophysical EM events.

  19. Orion EM-1 Booster Preps - Aft Skirt Preps/Painting

    NASA Image and Video Library

    2016-10-31

    The right hand aft skirt for NASA's Space Launch System (SLS) rocket has been refurbished and painted and is in a drying cell in a support building at the Hangar AF facility at Cape Canaveral Air Force Station in Florida. The space shuttle-era aft skirt will be used on the right hand booster of the SLS for Exploration Mission 1 (EM-1). NASA is preparing for EM-1, deep space missions, and the Journey to Mars.

  20. Orion EM-1 Booster Preps - Aft Skirt Preps/Painting

    NASA Image and Video Library

    2016-10-28

    A technician with Orbital ATK, prime contractor for the Space Launch System (SLS) Booster, preps a section of the right hand aft skirt for primer and paint in a support building at the Hangar AF facility at Cape Canaveral Air Force Station in Florida. The space shuttle-era aft skirt will be used on the right hand booster of NASA's SLS rocket for Exploration Mission 1 (EM-1). NASA is preparing for EM-1, deep space missions, and the Journey to Mars.

  1. Orion EM-1 Booster Preps - Aft Skirt Preps/Painting

    NASA Image and Video Library

    2016-10-29

    The right hand aft skirt for NASA's Space Launch System (SLS) rocket has been painted and is in a drying cell in a support building at the Hangar AF facility at Cape Canaveral Air Force Station in Florida. The space shuttle-era aft skirt will be used on the right hand booster of NASA's Space Launch System rocket for Exploration Mission 1 (EM-1). NASA is preparing for EM-1, deep space missions, and the Journey to Mars.

  2. EM Telemetry Tool for Deep Well Drilling Applications

    SciTech Connect

    Jeffrey M. Gabelmann

    2005-11-15

    This final report discusses the successful development and testing of a deep operational electromagnetic (EM) telemetry system, produced under a cooperative agreement with the United States Department of Energy's National Energy Technology Laboratory. This new electromagnetic telemetry system provides a wireless communication link between sensors deployed deep within oil and gas wells and data acquisition equipment located on the earth's surface. EM based wireless telemetry is a highly appropriate technology for oil and gas exploration in that it avoids the need for thousands of feet of wired connections. In order to achieve the project performance objectives, significant improvements over existing EM telemetry systems were made. These improvements included the development of new technologies that have improved the reliability of the communications link while extending operational depth. A key element of the new design is the incorporation of a data-fusion methodology which enhances the communication receiver's ability to extract very weak signals from large amounts of ambient environmental noise. This innovative data-fusion receiver based system adapts advanced technologies, not normally associated with low-frequency communications, and makes them work within the harsh drilling environments associated with the energy exploration market. Every element of a traditional EM telemetry system design, from power efficiency to reliability, has been addressed. The data fusion based EM telemetry system developed during this project is anticipated to provide an EM tool capability that will impact both onshore and offshore oil and gas exploration operations, for conventional and underbalanced drilling applications.

  3. STEME: efficient EM to find motifs in large data sets.

    PubMed

    Reid, John E; Wernisch, Lorenz

    2011-10-01

    MEME and many other popular motif finders use the expectation-maximization (EM) algorithm to optimize their parameters. Unfortunately, the running time of EM is linear in the length of the input sequences. This can prohibit its application to data sets of the size commonly generated by high-throughput biological techniques. A suffix tree is a data structure that can efficiently index a set of sequences. We describe an algorithm, Suffix Tree EM for Motif Elicitation (STEME), that approximates EM using suffix trees. To the best of our knowledge, this is the first application of suffix trees to EM. We provide an analysis of the expected running time of the algorithm and demonstrate that STEME runs an order of magnitude more quickly than the implementation of EM used by MEME. We give theoretical bounds for the quality of the approximation and show that, in practice, the approximation has a negligible effect on the outcome. We provide an open source implementation of the algorithm that we hope will be used to speed up existing and future motif search algorithms.

  4. STEME: efficient EM to find motifs in large data sets

    PubMed Central

    Reid, John E.; Wernisch, Lorenz

    2011-01-01

    MEME and many other popular motif finders use the expectation–maximization (EM) algorithm to optimize their parameters. Unfortunately, the running time of EM is linear in the length of the input sequences. This can prohibit its application to data sets of the size commonly generated by high-throughput biological techniques. A suffix tree is a data structure that can efficiently index a set of sequences. We describe an algorithm, Suffix Tree EM for Motif Elicitation (STEME), that approximates EM using suffix trees. To the best of our knowledge, this is the first application of suffix trees to EM. We provide an analysis of the expected running time of the algorithm and demonstrate that STEME runs an order of magnitude more quickly than the implementation of EM used by MEME. We give theoretical bounds for the quality of the approximation and show that, in practice, the approximation has a negligible effect on the outcome. We provide an open source implementation of the algorithm that we hope will be used to speed up existing and future motif search algorithms. PMID:21785132

  5. A History and Informal Assessment of the <em>Slacker Astronomyem> Podcast

    NASA Astrophysics Data System (ADS)

    Price, Aaron; Gay, Pamela; Searle, Travis; Brissenden, Gina

    Slacker Astronomyem> is a weekly podcast that covers a recent astronomical news event or discovery. The show has a unique style consisting of irreverent, over-the-top humor combined with a healthy dose of hard science. According to our demographic analysis, the combination of this style and the unique podcasting distribution mechanism allows the show to reach audiences younger and busier than those reached via traditional channels. We report on the successes and challenges of the first year of the show, and provide an informal assessment of its role as a source for astronomical news and concepts for its approximately 15,500 weekly listeners.

  6. A HF EM installation allowing simultaneous whole body and deep local EM hyperthermia.

    PubMed

    Mazokhin, V N; Kolmakov, D N; Lucheyov, N A; Gelvich, E A; Troshin, I I

    1999-01-01

    The structure and main features of a HF EM installation based upon a new approach for creating electromagnetic fields destined for whole body (WBH) and deep local (DLH) hyperthermia are discussed. The HF EM field, at a frequency of 13.56 MHz, is created by a coplanar capacity type applicator positioned under a distilled water filled bolus that the patient is lying on. The EM energy being released directly in the deep tissues ensures effective whole body heating to required therapeutic temperatures of up to 43.5 degrees C, whereas the skin temperature can be maintained as low as 39-40.5 degrees C. For DLH, the installation is equipped with additional applicators and a generator operating at a frequency of 40.68 MHz. High efficiency of the WBH applicator makes it possible to carry out the WBH procedure without any air-conditioning cabin. Due to this, a free access to the patient's body during the WBH treatment is provided and a simultaneous WBH/DLH or WBH/LH procedure by means of additional applicators is possible. Controllable power output in the range of 100-800 W at a frequency of 13.56 MHz and 50-350 W at a frequency of 40.68 MHz allows accurate temperature control during WBH, DLH and WBH/DLH procedures. SAR patterns created by the WBH and DLH applicators in a liquid muscle phantom and measured by means of a non-perturbing E-dipole are investigated. The scattered EM field strength measured in the vicinity of the operating installation during the WBH, DLH and WBH/DLH procedures does not exceed security standards. Examples of temperature versus time graphs in the course of WBH, DLH and WBH/DLH procedures in clinics are presented. The installation is successfully used in leading oncological institutions of Russia and Belarus, though combined WBH/DLH procedures are evidently more complicated and demand thorough planning and temperature measurements to avoid overheating.

  7. Airborne EM for mine infrastructure planning

    NASA Astrophysics Data System (ADS)

    Wijns, Chris

    2016-08-01

    Airborne electromagnetic (AEM) surveys with near-surface vertical resolution provide rapid and comprehensive coverage of a mine site ahead of infrastructure planning. In environments of sufficient electrical conductivity contrast, the data will map variations in the depth to bedrock, providing guidance for expected excavation depths for solid building foundations, or mine pre-strip volumes. Continuous coverage overcomes the severe areal limitation of relying only on drilling and test pits. An AEM survey in northern Finland illustrates the success of this approach for guiding the placement of a mine crusher and related infrastructure. The cost of the EM data collection and interpretation is insignificant in comparison to the US$300 million capital cost of the mine infrastructure. This environment of shallow glacial cover challenges the limits of AEM resolution, yet analysis of subsequently collected three-dimensional (3D) surface seismic data and actual pre-strip excavation depths reinforces the predictive, but qualitative, mapping capability of the AEM. It also highlights the need to tune the modelling via petrophysics for the specific goal of the investigation, and exposes the limitations of visual drill core logging.

  8. Databases and archiving for cryoEM

    PubMed Central

    Patwardhan, Ardan; Lawson, Catherine L.

    2017-01-01

    Cryo-EM in structural biology is currently served by three public archives – EMDB for 3DEM reconstructions, PDB for models built from 3DEM reconstructions and EMPIAR for the raw 2D image data used to obtain the 3DEM reconstructions. These archives play a vital role for both the structural community and the wider biological community in making the data accessible so that results may be reused, reassessed and integrated with other structural and bioinformatics resources. The important role of the archives is underpinned by the fact that many journals mandate the deposition of data to PDB and EMDB on publication. The field is currently undergoing transformative changes where on the one hand high-resolution structures are becoming a routine occurrence while on the other hand electron tomography is enabling the study of macromolecules in the cellular context. Concomitantly the archives are evolving to best serve their stakeholder communities. In this chapter we describe the current state of the archives, resources available for depositing, accessing, searching, visualising and validating data, on-going community-wide initiatives and opportunities and challenges for the future. PMID:27572735

  9. DOE EM industry programs robotics development

    SciTech Connect

    Staubly, R.; Kothari, V.

    1998-12-31

    The Office of Science and Technology (OST) manages an aggressive program for RD and D, as well as testing and evaluation for the Department of Energy`s (DOE`s) Environmental Management (EM) organization. The goal is to develop new and improved environmental restoration and waste management technologies to clean up the inventory of the DOE weapons complex faster, safer, and cheaper than is possible with currently available technologies. Robotic systems reduce worker exposure to the absolute minimum, while providing proven, cost-effective, and, for some applications, the only acceptable technique for addressing challenging problems. Development of robotic systems for remote operations occurs in three main categories: tank waste characterization and retrieval; decontamination and dismantlement; and characterization, mapping, and inspection systems. In addition, the Federal Energy Technology Center (FETC) has some other projects which fall under the heading of supporting R and D. The central objective of all FETC robotic projects is to make robotic systems more attractive by reducing costs and health risks associated with the deployment of robotic technologies in the cleanup of the nuclear weapons complex. This will be accomplished through development of robots that are cheaper, faster, safer, and more reliable, as well as more straightforward to modify/adapt and more intuitive to operate with autonomous capabilities and intelligent controls that prevent accidents and optimize task execution.

  10. Analysis of EM dataset with several sensor configurations obtained by the loop-loop EM survey on magnetic anomalies

    NASA Astrophysics Data System (ADS)

    CHOI, J.; Yi, M. J.; Sasaki, Y.; Son, J.; Nam, M. J.

    2015-12-01

    Most of mineral mines in Korea are located in rugged mountain area embedding small-scale anomalies. Loop-loop EM survey system can be a better choice for exploring those mines because no ground contact is required and portable loops are freely positioned. Survey design is very important for detecting small amount of mineral deposits efficiently and spatial limits of survey lines should be considered. Along a same survey line, surveys with different separations between a transmitter and a receiver are applicable. EM responses are calculated in a layered-earth model embedding magnetic anomalies and analyses considering electric conductivity and magnetic permeability are made for the loop-loop EM survey data. Combining EM dataset with multi-frequency and multi-separation slightly enhanced a reconstructed image. Loop-loop EM survey using PROMOIS system was conducted on a small magnetite mine. Inversion with and without considering magnetic permeability was conducted for EM data with multi-frequency and multi-separation between a transmitter and a receiver.

  11. Image segmentation by EM-based adaptive pulse coupled neural networks in brain magnetic resonance imaging.

    PubMed

    Fu, J C; Chen, C C; Chai, J W; Wong, S T C; Li, I C

    2010-06-01

    We propose an automatic hybrid image segmentation model that integrates the statistical expectation maximization (EM) model and the spatial pulse coupled neural network (PCNN) for brain magnetic resonance imaging (MRI) segmentation. In addition, an adaptive mechanism is developed to fine tune the PCNN parameters. The EM model serves two functions: evaluation of the PCNN image segmentation and adaptive adjustment of the PCNN parameters for optimal segmentation. To evaluate the performance of the adaptive EM-PCNN, we use it to segment MR brain image into gray matter (GM), white matter (WM) and cerebrospinal fluid (CSF). The performance of the adaptive EM-PCNN is compared with that of the non-adaptive EM-PCNN, EM, and Bias Corrected Fuzzy C-Means (BCFCM) algorithms. The result is four sets of boundaries for the GM and the brain parenchyma (GM+WM), the two regions of most interest in medical research and clinical applications. Each set of boundaries is compared with the golden standard to evaluate the segmentation performance. The adaptive EM-PCNN significantly outperforms the non-adaptive EM-PCNN, EM, and BCFCM algorithms in gray mater segmentation. In brain parenchyma segmentation, the adaptive EM-PCNN significantly outperforms the BCFCM only. However, the adaptive EM-PCNN is better than the non-adaptive EM-PCNN and EM on average. We conclude that of the three approaches, the adaptive EM-PCNN yields the best results for gray matter and brain parenchyma segmentation.

  12. FitEM2EM--tools for low resolution study of macromolecular assembly and dynamics.

    PubMed

    Frankenstein, Ziv; Sperling, Joseph; Sperling, Ruth; Eisenstein, Miriam

    2008-01-01

    Studies of the structure and dynamics of macromolecular assemblies often involve comparison of low resolution models obtained using different techniques such as electron microscopy or atomic force microscopy. We present new computational tools for comparing (matching) and docking of low resolution structures, based on shape complementarity. The matched or docked objects are represented by three dimensional grids where the value of each grid point depends on its position with regard to the interior, surface or exterior of the object. The grids are correlated using fast Fourier transformations producing either matches of related objects or docking models depending on the details of the grid representations. The procedures incorporate thickening and smoothing of the surfaces of the objects which effectively compensates for differences in the resolution of the matched/docked objects, circumventing the need for resolution modification. The presented matching tool FitEM2EMin successfully fitted electron microscopy structures obtained at different resolutions, different conformers of the same structure and partial structures, ranking correct matches at the top in every case. The differences between the grid representations of the matched objects can be used to study conformation differences or to characterize the size and shape of substructures. The presented low-to-low docking tool FitEM2EMout ranked the expected models at the top.

  13. Colloidal Oatmeal <em>(Avena Sativa)em> Improves Skin Barrier Through Multi-Therapy Activity.

    PubMed

    Ilnytska, Olha; Kaur, Simarna; Chon, Suhyoun; Reynertson, Kurt A; Nebus, Judith; Garay, Michelle; Mahmood, Khalid; Southall, Michael D

    2016-06-01

    Oats (Avena sativa) are a centuries-old topical treatment for a variety of skin barrier conditions, including dry skin, skin rashes, and eczema; however, few studies have investigated the actual mechanism of action for the skin barrier strengthening activity of colloidal oatmeal. Four extracts of colloidal oatmeal were prepared with various solvents and tested in vitro for skin barrier related gene expression and activity. Extracts of colloidal oatmeal were found to induce the expression of genes related to epidermal differentiation, tight junctions and lipid regulation in skin, and provide pH-buffering capacity. Colloidal oatmeal boosted the expression of multiple target genes related to skin barrier, and resulted in recovery of barrier damage in an in vitro model of atopic dermatitis. In addition, an investigator-blinded study was performed with 50 healthy female subjects who exhibited bilateral moderate to severe dry skin on their lower legs. Subjects were treated with a colloidal oatmeal skin protectant lotion. Clinically, the colloidal oatmeal lotion showed significant clinical improvements in skin dryness, moisturization, and barrier. Taken together, these results demonstrate that colloidal oatmeal can provide clinically effective benefits for dry and compromised skin by strengthening skin barrier.

    <em>J Drugs Dermatolem>. 2016;15(6):684-690.

  14. Emergency medical service (EMS): A unique flight environment

    NASA Technical Reports Server (NTRS)

    Shively, R. Jay

    1993-01-01

    The EMS flight environment is unique in today's aviation. The pilots must respond quickly to emergency events and often fly to landing zones where they have never been before . The time from initially receiving a call to being airborne can be as little as two to three minutes. Often the EMS pilot is the only aviation professional on site, they have no operations people or other pilots to aid them in making decisons. Further, since they are often flying to accident scenes, not airports, there is often complete weather and condition information. Therefore, the initial decision that the pilot must make, accepting or declining a flight, can become very difficult. The accident rate of EMS helicopters has been relatively high over the past years. NASA-Ames research center has taken several steps in an attempt to aid EMS pilots in their decision making and situational awareness. A preflight risk assessment system (SAFE) was developed to aid pilots in their decision making, and was tested at an EMS service. The resutls of the study were promising and a second version incorporating the lessons learned is under development. A second line of research was the development of a low cost electronic chart display (ECD). This is a digital map display to help pilots maintain geographical orientation. Another thrust was undertaken in conjunction with the Aviation Safety Reporting System (ASRS). This involved publicizing the ASRS to EMS pilots and personnel, and calling each of the reporters back to gather additional information. This paper will discuss these efforts and how they may positively impact the safety of EMS operations.

  15. Emergency medical service (EMS): A unique flight environment

    NASA Technical Reports Server (NTRS)

    Shively, R. Jay

    1993-01-01

    The EMS flight environment is unique in today's aviation. The pilots must respond quickly to emergency events and often fly to landing zones where they have never been before . The time from initially receiving a call to being airborne can be as little as two to three minutes. Often the EMS pilot is the only aviation professional on site, they have no operations people or other pilots to aid them in making decisons. Further, since they are often flying to accident scenes, not airports, there is often complete weather and condition information. Therefore, the initial decision that the pilot must make, accepting or declining a flight, can become very difficult. The accident rate of EMS helicopters has been relatively high over the past years. NASA-Ames research center has taken several steps in an attempt to aid EMS pilots in their decision making and situational awareness. A preflight risk assessment system (SAFE) was developed to aid pilots in their decision making, and was tested at an EMS service. The resutls of the study were promising and a second version incorporating the lessons learned is under development. A second line of research was the development of a low cost electronic chart display (ECD). This is a digital map display to help pilots maintain geographical orientation. Another thrust was undertaken in conjunction with the Aviation Safety Reporting System (ASRS). This involved publicizing the ASRS to EMS pilots and personnel, and calling each of the reporters back to gather additional information. This paper will discuss these efforts and how they may positively impact the safety of EMS operations.

  16. Degradation of Benzodiazepines after 120 Days of EMS Deployment

    PubMed Central

    McMullan, Jason T.; Jones, Elizabeth; Barnhart, Bruce; Denninghoff, Kurt; Spaite, Daniel; Zaleski, Erin; Silbergleit, Robert

    2014-01-01

    Introduction EMS treatment of status epilepticus improves outcomes, but the benzodiazepine best suited for EMS use is unclear, given potential high environmental temperature exposures. Objective To describe the degradation of diazepam, lorazepam, and midazolam as a function of temperature exposure and time over 120 days of storage on active EMS units. Methods Study boxes containing vials of diazepam, lorazepam, and midazolam were distributed to 4 active EMS units in each of 2 EMS systems in the southwestern United States during May–August 2011. The boxes logged temperature every minute and were stored in EMS units per local agency policy. Two vials of each drug were removed from each box at 30-day intervals and underwent high-performance liquid chromatography to determine drug concentration. Concentration was analyzed as mean (and 95%CI) percent of initial labeled concentration as a function of time and mean kinetic temperature (MKT). Results 192 samples were collected (2 samples of each drug from each of 4 units per city at 4 time-points). After 120 days, the mean relative concentration (95%CI) of diazepam was 97.0% (95.7–98.2%) and of midazolam was 99.0% (97.7–100.2%). Lorazepam experienced modest degradation by 60 days (95.6% [91.6–99.5%]) and substantial degradation at 90 days (90.3% [85.2-95.4%]) and 120 days (86.5% [80.7–92.3%]). Mean MKT was 31.6°C (95%CI 27.1–36.1). Increasing MKT was associated with greater degradation of lorazepam, but not midazolam or diazepam. Conclusions Midazolam and diazepam experienced minimal degradation throughout 120 days of EMS deployment in high-heat environments. Lorazepam experienced significant degradation over 120 days and appeared especially sensitive to higher MKT exposure. PMID:24548058

  17. Resource Document: Coordination of Pediatric Emergency Care in EMS Systems.

    PubMed

    Remick, Katherine; Gross, Toni; Adelgais, Kathleen; Shah, Manish I; Leonard, Julie C; Gausche-Hill, Marianne

    2017-01-01

    Citing numerous pediatric-specific deficiencies within Emergency Medical Services (EMS) systems, the Institute of Medicine (IOM) recommended that EMS systems appoint a pediatric emergency care coordinator (PECC) to provide oversight of EMS activities related to care of children, to promote the integration of pediatric elements into day-to-day services as well as local and/or regional disaster planning, and to promote pediatric education across all levels of EMS providers. A systematic review of the literature was undertaken to describe the evidence for pediatric coordination across the emergency care continuum. The search strategy was developed by the investigators in consultation with a medical librarian and conducted in OVID, Medline, PubMed, Embase, Web of Science, and CINAHL databases from January 1, 1983 to January 1, 2016. All research articles that measured a patient-related or system-related outcome associated with pediatric coordination in the setting of emergency care, trauma, or disaster were included. Opinion articles, commentaries, and letters to the editors were excluded. Three investigators independently screened citations in a hierarchical manner and abstracted data. Of 149 identified titles, nine were included in the systematic review. The nine articles included one interventional study, five surveys, and three consensus documents. All articles favored the presence of pediatric coordination. The interventional study demonstrated improved documentation, clinical management, and staff awareness of high priority pediatric areas. The current literature supports the identification of pediatric coordination to facilitate the optimal care of children within EMS systems. In order for EMS systems to provide high quality care to children, pediatric components must be integrated into all aspects of care including day-to-day operations, policies, protocols, available equipment and medications, quality improvement efforts, and disaster planning. This systematic

  18. The EM-POGO: A simple, absolute velocity profiler

    NASA Astrophysics Data System (ADS)

    Terker, S. R.; Sanford, T. B.; Dunlap, J. H.; Girton, J. B.

    2013-01-01

    Electromagnetic current instrumentation has been added to the Bathy Systems, Inc. POGO transport sondes to produce a free-falling absolute velocity profiler called EM-POGO. The POGO is a free-fall profiler that measures a depth-averaged velocity using GPS fixes at the beginning and end of a round trip to the ocean floor (or a preset depth). The EM-POGO adds a velocity profile determined from measurements of motionally induced electric fields generated by the ocean current moving through the vertical component of the Earth's magnetic field. In addition to providing information about the vertical structure of the velocity, the depth-dependent measurements improve transport measurements by correcting for the non-constant fall-rate. Neglecting the variable fall rate results in errors O (1 cm s-1). The transition from POGO to EM-POGO included electrically isolating the POGO and electric-field-measuring circuits, installing a functional GPS receiver, finding a pressure case that provided an optimal balance among crush-depth, price and size, and incorporating the electrodes, electrode collar, and the circuitry required for the electric field measurement. The first EM-POGO sea-trial was in July 1999. In August 2006 a refurbished EM-POGO collected 15 absolute velocity profiles; relative and absolute velocity uncertainty was ˜1cms-1 and 0.5-5 cm s-1, respectively, at a vertical resolution of 25 m. Absolute velocity from the EM-POGO compared to shipboard ADCP measurements differed by ˜ 1-2 cm s-1, comparable to the uncertainty in absolute velocity from the ADCP. The EM-POGO is thus a low-cost, easy to deploy and recover, and accurate velocity profiler.

  19. International Space Station (ISS) Emergency Mask (EM) Development

    NASA Technical Reports Server (NTRS)

    Toon, Katherine P.; Hahn, Jeffrey; Fowler, Michael; Young, Kevin

    2011-01-01

    The Emergency Mask (EM) is considered a secondary response emergency Personal Protective Equipment (PPE) designed to provide respiratory protection to the International Space Station (ISS) crewmembers in response to a post-fire event or ammonia leak. The EM is planned to be delivered to ISS in 2012 to replace the current air purifying respirator (APR) onboard ISS called the Ammonia Respirator (AR). The EM is a one ]size ]fits ]all model designed to fit any size crewmember, unlike the APR on ISS, and uses either two Fire Cartridges (FCs) or two Commercial Off-the-Shelf (COTS) 3M(Trademark). Ammonia Cartridges (ACs) to provide the crew with a minimum of 8 hours of respiratory protection with appropriate cartridge swap ]out. The EM is designed for a single exposure event, for either post ]fire or ammonia, and is a passive device that cannot help crewmembers who cannot breathe on their own. The EM fs primary and only seal is around the wearer fs neck to prevent a crewmember from inhaling contaminants. During the development of the ISS Emergency Mask, several design challenges were faced that focused around manufacturing a leak free mask. The description of those challenges are broadly discussed but focuses on one key design challenge area: bonding EPDM gasket material to Gore(Registered Trademark) fabric hood.

  20. Breaking Cryo-EM Resolution Barriers to Facilitate Drug Discovery.

    PubMed

    Merk, Alan; Bartesaghi, Alberto; Banerjee, Soojay; Falconieri, Veronica; Rao, Prashant; Davis, Mindy I; Pragani, Rajan; Boxer, Matthew B; Earl, Lesley A; Milne, Jacqueline L S; Subramaniam, Sriram

    2016-06-16

    Recent advances in single-particle cryoelecton microscopy (cryo-EM) are enabling generation of numerous near-atomic resolution structures for well-ordered protein complexes with sizes ≥ ∼200 kDa. Whether cryo-EM methods are equally useful for high-resolution structural analysis of smaller, dynamic protein complexes such as those involved in cellular metabolism remains an important question. Here, we present 3.8 Å resolution cryo-EM structures of the cancer target isocitrate dehydrogenase (93 kDa) and identify the nature of conformational changes induced by binding of the allosteric small-molecule inhibitor ML309. We also report 2.8-Å- and 1.8-Å-resolution structures of lactate dehydrogenase (145 kDa) and glutamate dehydrogenase (334 kDa), respectively. With these results, two perceived barriers in single-particle cryo-EM are overcome: (1) crossing 2 Å resolution and (2) obtaining structures of proteins with sizes < 100 kDa, demonstrating that cryo-EM can be used to investigate a broad spectrum of drug-target interactions and dynamic conformational states.

  1. Refinement of Atomic Structures Against cryo-EM Maps.

    PubMed

    Murshudov, G N

    2016-01-01

    This review describes some of the methods for atomic structure refinement (fitting) against medium/high-resolution single-particle cryo-EM reconstructed maps. Some of the tools developed for macromolecular X-ray crystal structure analysis, especially those encapsulating prior chemical and structural information can be transferred directly for fitting into cryo-EM maps. However, despite the similarities, there are significant differences between data produced by these two techniques; therefore, different likelihood functions linking the data and model must be used in cryo-EM and crystallographic refinement. Although tools described in this review are mostly designed for medium/high-resolution maps, if maps have sufficiently good quality, then these tools can also be used at moderately low resolution, as shown in one example. In addition, the use of several popular crystallographic methods is strongly discouraged in cryo-EM refinement, such as 2Fo-Fc maps, solvent flattening, and feature-enhanced maps (FEMs) for visualization and model (re)building. Two problems in the cryo-EM field are overclaiming resolution and severe map oversharpening. Both of these should be avoided; if data of higher resolution than the signal are used, then overfitting of model parameters into the noise is unavoidable, and if maps are oversharpened, then at least parts of the maps might become very noisy and ultimately uninterpretable. Both of these may result in suboptimal and even misleading atomic models.

  2. Application of the EM algorithm to radiographic images.

    PubMed

    Brailean, J C; Little, D; Giger, M L; Chen, C T; Sullivan, B J

    1992-01-01

    The expectation maximization (EM) algorithm has received considerable attention in the area of positron emitted tomography (PET) as a restoration and reconstruction technique. In this paper, the restoration capabilities of the EM algorithm when applied to radiographic images is investigated. This application does not involve reconstruction. The performance of the EM algorithm is quantitatively evaluated using a "perceived" signal-to-noise ratio (SNR) as the image quality metric. This perceived SNR is based on statistical decision theory and includes both the observer's visual response function and a noise component internal to the eye-brain system. For a variety of processing parameters, the relative SNR (ratio of the processed SNR to the original SNR) is calculated and used as a metric to compare quantitatively the effects of the EM algorithm with two other image enhancement techniques: global contrast enhancement (windowing) and unsharp mask filtering. The results suggest that the EM algorithm's performance is superior when compared to unsharp mask filtering and global contrast enhancement for radiographic images which contain objects smaller than 4 mm.

  3. Topical Treatment With an Agent Disruptive to <em>P. acnesem> Biofilm Provides Positive Therapeutic Response: Results of a Randomized Clinical Trial.

    PubMed

    Bernhardt, Michael J; Myntti, Matthew F

    2016-06-01

    The traditional disease model of acne has been one of follicular plugging due to 'sticky epithelial cells' associated with increased sebum production with deep follicular anaerobic conditions favoring <em>P. acnesem>- generated inflammation. <em>P. acnesem> biofilms have been found more frequently in patients with acne than controls. Biofilms are genetically coded to create adhesion to the pilosebaceous unit followed by production of a mucopolysaccharide coating capable of binding to lipid surfaces. Traditional therapies for acne have involved mixtures of oral and topical antibiotics admixed with topical keratolytics and retinoids, which are aimed at traditional bacterial reduction as well as downregulating the inflammatory cascade. These approaches are limited by side effect and compliance/tolerability issues. As the <em>P. acnesem> biofilm may, in fact, be the instigator of this process, we studied the use of a topical agent designed to reduce the <em>P. acnesem> biofilm to see if reducing the biofilm would be therapeutically efficacious. We present data of a proprietary topical non-prescription agent with a novel pharmaco mechanism designed to attack the biofilm produced by <em>P. acnesem>. Our data shows a decrease of inflammatory lesions by 44% and non-inflammatory lesions by 32% after 12 weeks and also provided for a meaningful improvement in the quality of life of the patients in the study. These improvements were achieved with a product that was not associated with burning, chafing, irritation, or erythema, which can be seen with topical treatments. It is apparent from this study that by addressing the biofilm which protects the <em>P. acnesem> bacteria through the use of the Acne Gel, the incidence of acne symptoms can be greatly reduced, while having no negative impacts on the patients' skin (ClinicalTrials.gov registry number NCT02404285).

    <em>J Drugs Dermatol. em>2016;15(6):677-683.

  4. [Epidemiological and Demographic Characteristics of Patients with Head and Neck Tumours in the Northern Portugal: Impact on Survival].

    PubMed

    Estêvão, Roberto; Santos, Tiago; Ferreira, Ana; Machado, Anabela; Fernandes, João; Monteiro, Eurico

    2016-10-01

    Introdução: Em Portugal não existem, ainda, estudos epidemiológicos e demográficos realizados em doentes portadores de tumores da cabeça e pescoço. Os objectivos desta análise são descrever os aspectos epidemiológicos e demográficos dos doentes portadores de tumores malignos da cabeça e do pescoço, referenciados ao serviço de Otorrinolaringologia de um centro oncológico do norte de Portugal e avaliar o impacto destas características na sobrevivência. Material e Métodos: Estudo retrospectivo descritivo, dos doentes referenciados entre janeiro de 2011 e dezembro de 2013 ao Serviço de Otorrinolaringologia do Instituto Português de Oncologia do Porto. Foram incluídos 566 doentes. As variáveis analisadas foram a localização anatómica dos tumores, o estadiamento, o género e a idade dos doentes, o distrito de proveniência, os hábitos etílicos e tabágicos, o nível educacional, a profissão, o estado civil e a estrutura familiar.Resultados: Os 566 doentes estudados correspondem a 498 homens e 68 mulheres, com idade média de 58,1 ± 12,2 anos. A maioria (80,5%) foi referenciada com doença em estadio avançado (III e IV). Verificamos hábitos etílicos moderados a excessivos em 78% dos doentes e 69% eram fumadores. A grande maioria dos doentes (82,3%) tinha apenas o ensino básico. Os doentes com tumores da hipofaringe e com hábitos etílicos excessivos foram referenciados em estadios mais avançados e apresentaram pior sobrevivência (p < 0,001). Os doentes casados foram referenciados em estadios mais precoces (p = 0,020) e os doentes sem apoio familiar apresentaram pior sobrevivência (p = 0,030). Discussão: Os dados epidemiológicos encontrados estão de acordo com a literatura internacional. A elevada taxa de doentes referenciados em estádio avançado pode ser atribuída ao atraso na procura de cuidados médicos ou a factores inerentes ao sistema de referenciação. A vigilância e o suporte familiar têm um papel

  5. Where There is No EMS: Lay Providers in Emergency Medical Services Care - EMS as a Public Health Priority.

    PubMed

    Debenham, Sierra; Fuller, Matthew; Stewart, Matthew; Price, Raymond R

    2017-08-11

    By 2030, road traffic accidents are projected to be the fifth leading cause of death worldwide, with 90% of these deaths occurring in low- and middle-income countries (LMICs). While high-quality, prehospital trauma care is crucial to reduce the number of trauma-related deaths, effective Emergency Medical Systems (EMS) are limited or absent in many LMICs. Although lay providers have long been recognized as the front lines of informal trauma care in countries without formal EMS, few efforts have been made to capitalize on these networks. We suggest that lay providers can become a strong foundation for nascent EMS through a four-fold approach: strengthening and expanding existing lay provider training programs; incentivizing lay providers; strengthening locally available first aid supply chains; and using technology to link lay provider networks. Debenham S , Fuller M , Stewart M , Price RR . Where there is no EMS: lay providers in Emergency Medical Services care - EMS as a public health priority. Prehosp Disaster Med. 2017;32(6):1-3.

  6. EMS Stretcher “Misadventures” in a Large, Urban EMS System: A Descriptive Analysis of Contributing Factors and Resultant Injuries

    PubMed Central

    Goodloe, Jeffrey M.; Crowder, Christopher J.; Arthur, Annette O.; Thomas, Stephen H.

    2012-01-01

    Purpose. There is a paucity of data regarding EMS stretcher-operation-related injuries. This study describes and analyzes characteristics associated with undesirable stretcher operations, with or without resultant injury in a large, urban EMS agency. Methods. In the study agency, all stretcher-related “misadventures” are required to be documented, regardless of whether injury results. All stretcher-related reports between July 1, 2009 and June 30, 2010 were queried in retrospective analysis, avoiding Hawthorne effect in stretcher operations. Results. During the year studied, 129,110 patients were transported. 23 stretcher incidents were reported (0.16 per 1,000 transports). No patient injury occurred. Four EMS providers sustained minor injuries. Among contributing aspects, the most common involved operations surrounding the stretcher-ambulance safety latch, 14/23 (60.9%). From a personnel injury prevention perspective, there exists a significant relationship between combative patients and crew injury related to stretcher operation, Fisher's exact test 0.048. Conclusions. In this large, urban EMS system, the incidence of injury related to stretcher operations in the one-year study period is markedly low, with few personnel injuries and no patient injuries incurred. Safety for EMS personnel and patients could be advanced by educational initiatives that highlight specific events and conditions contributing to stretcher-related adverse events. PMID:22606379

  7. EMS-induced cytomictic variability in safflower (Carthamus tinctorius L.).

    PubMed

    Srivastava, P; Kumar, G

    2011-01-01

    Seeds of safflower (Carthamus tinctorius L.) were subjected to three treatment durations (3h, 5h and 7h) of 0.5 % Ethyl Methane Sulphonate (EMS). Microsporogenesis was carried out in the control as well as in the treated materials. EMS treated plants showed interesting feature of partial inter-meiocyte chromatin migration through channel formation, beak formation or direct cell fusion. Another interesting feature noticed during the study was the fusion among tetrads due to wall dissolution. The phenomenon of cytomixis was recorded at nearly all the stages of microsporogenesis connecting from a few to several meiocytes. Other abnormalities such as laggards, precocious movement, bridge and non-disjunction of chromosomes were also recorded but in very low frequencies. The phenomenon of cytomixis increased along with the increase in treatment duration of EMS. Cells with these types of cytomictic disturbances may probably result in uneven formation of gametes or zygote, heterogenous sized pollen grains or even loss of fertility in future.

  8. Pirmasis lietuvi\\vskas Suvalkijos žemėlapis

    NASA Astrophysics Data System (ADS)

    Girkus, Romualdas; Lukoševičius, Viktoras

    2009-01-01

    Po ilgo paie\\vskos laikotarpio, dr. N. Lietuvninkaitei padedant, KTU bibliotekos Retų knygų skyriaus kartografiniame archyve aptiktas 1915 m. JAV išleistas B. K. Balučio M 1:252 000 Suvalkijos žemėlapis. Lietuvių išeivijos instituto B. K. Balučio fonde žemėlapio sudarymo medžiagos nerasta. Paie\\vska atrodė bevilti\\vska dėl tarpukario Lietuvi\\vskosios enciklopedijos klaidinančios informacijos, kad prasidėjus Pirmajam pasauliniam karui žemėlapis nebuvo išleistas, tad aprašomas ir publikuojamas pirmą kartą.

  9. Recent technical advancements enabled atomic resolution CryoEM

    NASA Astrophysics Data System (ADS)

    Xueming, Li

    2016-01-01

    With recent breakthroughs in camera and image processing technologies single-particle electron cryo-microscopy (CryoEM) has suddenly gained the attention of structural biologists as a powerful tool able to solve the atomic structures of biological complexes and assemblies. Compared with x-ray crystallography, CryoEM can be applied to partially flexible structures in solution and without the necessity of crystallization, which is especially important for large complexes and assemblies. This review briefly explains several key bottlenecks for atomic resolution CryoEM, and describes the corresponding solutions for these bottlenecks based on the recent technical advancements. The review also aims to provide an overview about the technical differences between its applications in biology and those in material science. Project supported by Tsinghua-Peking Joint Center for Life Sciences, China.

  10. Analyses of Subnanometer Resolution Cryo-EM Density Maps

    PubMed Central

    Baker, Matthew L.; Baker, Mariah R.; Hryc, Corey F.; DiMaio, Frank

    2011-01-01

    Today, electron cryomicroscopy (cryo-EM) can routinely achieve subnanometer resolutions of complex macromolecular assemblies. From a density map, one can extract key structural and functional information using a variety of computational analysis tools. At subnanometer resolution, these tools make it possible to isolate individual subunits, identify secondary structures, and accurately fit atomic models. With several cryo-EM studies achieving resolutions beyond 5 Å, computational modeling and feature recognition tools have been employed to construct backbone and atomic models of the protein components directly from a density map. In this chapter, we describe several common classes of computational tools that can be used to analyze and model subnanometer resolution reconstructions from cryo-EM. A general protocol for analyzing subnanometer resolution density maps is presented along with a full description of steps used in analyzing the 4.3 Å resolution structure of Mm-cpn. PMID:20888467

  11. Lietuvos Teritorijos Gravimetrinio Žemėlapio Tikslumo Vertinimas

    NASA Astrophysics Data System (ADS)

    Birvydienė, Rosita; Krikštaponis, Boleslovas; Obuchovski, Romuald; Paršeliūnas, Eimuntas; Petroškevičius, Petras; Šlikas, Dominykas

    2010-01-01

    Remiantis matavimų, atliktų Lietuvos gravimetrinio tinklo punktuose, duomenimis, įvertintas Lietuvos teritorijos 1:200 000 mastelio Bouguer anomalijų gravimetrinio žemėlapio tikslumas. Vertinimui naudota 686 gravimetrinių punktų sunkio pagreičio reikšmės, kurių vidutinė kvadratinė paklaida neviršija 9 μGal. Nustatytos teritorijos, kuriose sunkio pagreitis, apskaičiuotas pagal žemėlapį, turi sistemingąsias paklaidas. Gautas vidutinis skirtumas tarp žemėlapyje taikomos Potsdamo sunkio sistemos ir IGSN 71 sistemos, apibrėžtos Lietuvos gravimetriniame tinkle atliktais absoliučiaisiais gravimetriniais matavimais.

  12. Longitudinal information and radiation damage in EM calorimetry

    SciTech Connect

    Green, D.

    1993-02-05

    The SCC radiation field is higher than that encountered by previous hadron collider detectors. In particular, the electromagnetic (EM) calorimeter compartment sees the highest radiation dose. Since an EM calorimeter also makes the most precise energy measurement, special care must be lavished on this part of a calorimeter. Previous studies have concentrated on Monte Carlo examinations of 2 longitudinal compartments within the EM which can alleviate radiation damage. Recently, it was realized that a ``shower maximum`` detector, such as exists in CDF, also contains information of the conversion point of an electromagnetic shower. As such, it can potentially be used in a fashion analogous to the longitudinal compartments, although it is not designed to be optimized for this role.

  13. Geospatial Analysis of Pediatric EMS Run Density and Endotracheal Intubation

    PubMed Central

    Hansen, Matthew; Loker, William; Warden, Craig

    2016-01-01

    Introduction The association between geographic factors, including transport distance, and pediatric emergency medical services (EMS) run clustering on out-of-hospital pediatric endotracheal intubation is unclear. The objective of this study was to determine if endotracheal intubation procedures are more likely to occur at greater distances from the hospital and near clusters of pediatric calls. Methods This was a retrospective observational study including all EMS runs for patients less than 18 years of age from 2008 to 2014 in a geographically large and diverse Oregon county that includes densely populated urban areas near Portland and remote rural areas. We geocoded scene addresses using the automated address locator created in the cloud-based mapping platform ArcGIS, supplemented with manual address geocoding for remaining cases. We then use the Getis-Ord Gi spatial statistic feature in ArcGIS to map statistically significant spatial clusters (hot spots) of pediatric EMS runs throughout the county. We then superimposed all intubation procedures performed during the study period on maps of pediatric EMS-run hot spots, pediatric population density, fire stations, and hospitals. We also performed multivariable logistic regression to determine if distance traveled to the hospital was associated with intubation after controlling for several confounding variables. Results We identified a total of 7,797 pediatric EMS runs during the study period and 38 endotracheal intubations. In univariate analysis we found that patients who were intubated were similar to those who were not in gender and whether or not they were transported to a children’s hospital. Intubated patients tended to be transported shorter distances and were older than non-intubated patients. Increased distance from the hospital was associated with reduced odds of intubation after controlling for age, sex, scene location, and trauma system entry status in a multivariate logistic regression. The

  14. The photon: EM fields, electrical potentials, and AC charge

    NASA Astrophysics Data System (ADS)

    Meulenberg, A.; Hudgins, W. R.; Penland, R. F.

    2015-09-01

    Photons are here considered to be resonant oscillations (solitons) in four dimensions (space/time) of an undefined `field' otherwise generally existing at a local energy minimum. The photons' constituent EM fields result in elevated energy, and therefore potentials, within that field. It is in the context of the standing waves of and between photons that the EM fields and potentials lead to a description of alternating (AC) `currents' (of some form) of unquantized alternating `charge' (of some sort). The main topic of this paper is the alternating charge.

  15. Geospatial Analysis of Pediatric EMS Run Density and Endotracheal Intubation.

    PubMed

    Hansen, Matthew; Loker, William; Warden, Craig

    2016-09-01

    The association between geographic factors, including transport distance, and pediatric emergency medical services (EMS) run clustering on out-of-hospital pediatric endotracheal intubation is unclear. The objective of this study was to determine if endotracheal intubation procedures are more likely to occur at greater distances from the hospital and near clusters of pediatric calls. This was a retrospective observational study including all EMS runs for patients less than 18 years of age from 2008 to 2014 in a geographically large and diverse Oregon county that includes densely populated urban areas near Portland and remote rural areas. We geocoded scene addresses using the automated address locator created in the cloud-based mapping platform ArcGIS, supplemented with manual address geocoding for remaining cases. We then use the Getis-Ord Gi spatial statistic feature in ArcGIS to map statistically significant spatial clusters (hot spots) of pediatric EMS runs throughout the county. We then superimposed all intubation procedures performed during the study period on maps of pediatric EMS-run hot spots, pediatric population density, fire stations, and hospitals. We also performed multivariable logistic regression to determine if distance traveled to the hospital was associated with intubation after controlling for several confounding variables. We identified a total of 7,797 pediatric EMS runs during the study period and 38 endotracheal intubations. In univariate analysis we found that patients who were intubated were similar to those who were not in gender and whether or not they were transported to a children's hospital. Intubated patients tended to be transported shorter distances and were older than non-intubated patients. Increased distance from the hospital was associated with reduced odds of intubation after controlling for age, sex, scene location, and trauma system entry status in a multivariate logistic regression. The locations of intubations were

  16. A constrained EM algorithm for independent component analysis.

    PubMed

    Welling, M; Weber, M

    2001-03-01

    We introduce a novel way of performing independent component analysis using a constrained version of the expectation-maximization (EM) algorithm. The source distributions are modeled as D one-dimensional mixtures of gaussians. The observed data are modeled as linear mixtures of the sources with additive, isotropic noise. This generative model is fit to the data using constrained EM. The simpler "soft-switching" approach is introduced, which uses only one parameter to decide on the sub- or supergaussian nature of the sources. We explain how our approach relates to independent factor analysis.

  17. Low Bandwidth Vocoding using EM Sensor and Acoustic Signal Processing

    SciTech Connect

    Ng, L C; Holzrichter, J F; Larson, P E

    2001-10-25

    Low-power EM radar-like sensors have made it possible to measure properties of the human speech production system in real-time, without acoustic interference [1]. By combining these data with the corresponding acoustic signal, we've demonstrated an almost 10-fold bandwidth reduction in speech compression, compared to a standard 2.4 kbps LPC10 protocol used in the STU-III (Secure Terminal Unit, third generation) telephone. This paper describes a potential EM sensor/acoustic based vocoder implementation.

  18. Speaker verification using combined acoustic and EM sensor signal processing

    SciTech Connect

    Ng, L C; Gable, T J; Holzrichter, J F

    2000-11-10

    Low Power EM radar-like sensors have made it possible to measure properties of the human speech production system in real-time, without acoustic interference. This greatly enhances the quality and quantity of information for many speech related applications. See Holzrichter, Burnett, Ng, and Lea, J. Acoustic. SOC. Am . 103 ( 1) 622 (1998). By combining the Glottal-EM-Sensor (GEMS) with the Acoustic-signals, we've demonstrated an almost 10 fold reduction in error rates from a speaker verification system experiment under a moderate noisy environment (-10dB).

  19. Orion EM-1 Booster Preps - Aft Skirt Preps/Painting

    NASA Image and Video Library

    2016-10-28

    A paint technician with Orbital ATK, prime contractor for the Space Launch System (SLS) Booster, uses an air gun to apply paint to the right hand aft skirt for NASA’s SLS rocket inside a support building at the Hangar AF facility at Cape Canaveral Air Force Station. The space shuttle-era aft skirt, was inspected and resurfaced to prepare it for primer and paint. The aft skirt will be used on the right hand booster of the SLS rocket for Exploration Mission 1 (EM-1). NASA is preparing for EM-1, deep-space missions, and the journey to Mars.

  20. Orion EM-1 Booster Preps - Aft Skirt Preps/Painting

    NASA Image and Video Library

    2016-10-28

    Technicians with Orbital ATK, prime contractor for the Space Launch System (SLS) Booster, prepare the right hand aft skirt for NASA’s SLS rocket for primer and painting inside a support building at the Hangar AF facility at Cape Canaveral Air Force Station in Florida. The space shuttle-era aft skirt, was inspected and resurfaced and will be primed and painted for use on the right hand booster of the SLS rocket for Exploration Mission 1 (EM-1). NASA is preparing for EM-1, deep-space missions, and the journey to Mars.

  1. Orion EM-1 Booster Preps - Aft Skirt Preps/Painting

    NASA Image and Video Library

    2016-10-28

    Technicians with Orbital ATK, prime contractor for the Space Launch System (SLS) Booster, prepare a paint mixture for the right hand aft skirt for NASA’s SLS in a support building at the Hangar AF facility at Cape Canaveral Air Force Station in Florida. The space shuttle-era aft skirt, was inspected and resurfaced, and will be primed and painted for use on the right hand booster of the SLS rocket for Exploration Mission 1 (EM-1). NASA is preparing for EM-1, deep-space missions, and the Journey to Mars.

  2. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA's Kennedy Space Center in Florida, Lockheed Martin technicians secure a protective cover around the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) for its move to a clean room. The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  3. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA’s Kennedy Space Center in Florida, Lockheed Martin technicians move the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) into a clean room. The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  4. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA’s Kennedy Space Center in Florida, the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) is in a clean room with protective walls secured around it. The adapter will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  5. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA’s Kennedy Space Center in Florida, Lockheed Martin technicians move the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) toward a clean room. The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  6. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA’s Kennedy Space Center in Florida, a Lockheed Martin technician secures a protective cover around the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) for its move to a clean room The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  7. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA’s Kennedy Space Center in Florida, the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) is being moved to a clean room. The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  8. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA’s Kennedy Space Center in Florida, a protective cover is installed around the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) for its move to a clean room. The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  9. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA’s Kennedy Space Center in Florida, Lockheed Martin technicians secure a protective cover around the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) for its move to a clean room. The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  10. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA’s Kennedy Space Center in Florida, Lockheed Martin technicians are preparing the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) for the move into a clean room. The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  11. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA's Kennedy Space Center in Florida, Lockheed Martin technicians secure a protective cover around the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) for its move to a clean. The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  12. Orion EM-1 Crew Module Adapter Move to Clean Room

    NASA Image and Video Library

    2016-11-29

    Inside the Neil Armstrong Operations and Checkout Building high bay at NASA’s Kennedy Space Center in Florida, Lockheed Martin technicians begin to move the Orion crew module adapter (CMA) for Exploration Mission 1 (EM-1) to a clean room. The CMA will undergo propellant and environmental control and life support system tube installation and welding. The adapter will connect the Orion crew module to the European Space Agency-provided service module. The Orion spacecraft will launch atop NASA’s Space Launch System rocket on EM-1, its first deep space mission, in late 2018.

  13. A emissão em 8mm e as bandas de Merrill-Sanford em estrelas carbonadas

    NASA Astrophysics Data System (ADS)

    de Mello, A. B.; Lorenz-Martins, S.

    2003-08-01

    Estrelas carbonadas possuem bandas moleculares em absorção no visível e, no infravermelho (IR) as principais características espectrais se devem a emissão de grãos. Recentemente foi detectada a presença de bandas de SiC2 (Merrill-Sanford, MS) em emissão sendo atribuída à presença de um disco rico em poeira. Neste trabalho analisamos uma amostra de 14 estrelas carbonadas, observadas no telescópio de 1.52 m do ESO em 4 regiões espectrais diferentes, a fim de detectar as bandas de MS em emissão. Nossa amostra é composta de estrelas que apresentam além da emissão em 11.3 mm, outra em 8 mm. Esta última emissão, não usual nestes objetos, tem sido atribuída ou a moléculas de C2H2, ou a um composto sólido ainda indefinido. A detecção de emissões de MS e aquelas no IR, simultaneamente, revelaria um cenário mais complexo que o habitualmente esperado para os ventos destes objetos. No entanto como primeiro resultado, verificamos que as bandas de Merrill-Sanford encontram-se em absorção, não revelando nenhuma conexão com a emissão a 8 mm. Assim, temos duas hipóteses: (a) a emissão a 8 mm se deve à molécula C2H2 ou (b) essa emissão é resultado da emissão térmica de grãos. Testamos a segunda hipótese modelando a amostra com grãos não-homogêneos de SiC e quartzo, o qual emite em aproximadamente 8mm. Este grão seria produzido em uma fase evolutiva anterior a das carbonadas (estrelas S) e por terem uma estrutura cristalina são destruídos apenas na presença de campos de radiação ultravioleta muito intensos. Os modelos para os envoltórios utilizam o método de Monte Carlo para descrever o problema do transporte da radiação. As conclusões deste trabalho são: (1) as bandas de Merrill-Sanford se encontram em absorção, sugerindo um cenário usual para os ventos das estrelas da amostra; (2) neste cenário, a emissão em 8 mm seria resultado de grãos de quartzo com mantos de SiC, indicando que o quartzo poderia sobreviver a fase

  14. Item Parameter Estimation via Marginal Maximum Likelihood and an EM Algorithm: A Didactic.

    ERIC Educational Resources Information Center

    Harwell, Michael R.; And Others

    1988-01-01

    The Bock and Aitkin Marginal Maximum Likelihood/EM (MML/EM) approach to item parameter estimation is an alternative to the classical joint maximum likelihood procedure of item response theory. This paper provides the essential mathematical details of a MML/EM solution and shows its use in obtaining consistent item parameter estimates. (TJH)

  15. An EM System with Dynamic Multi-Axis Transmitter and Tensor Gradiometer Receiver

    DTIC Science & Technology

    2011-06-01

    FINAL REPORT An EM System With Dynamic Multi-Axis Transmitter and Tensor Gradiometer Receiver SERDP Project MR-1534 JUNE 2011 David C...2006- 2010 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER An EM System with Dynamic Multi-Axis transmitter and Tensor Gradiometer W91 2HQ-06-C-0050...239.18 EM Tensor Gradiometer SERDP MM-1532 i Contents Contents

  16. Causes of Death in an Acute Psychiatric Inpatient Unit of a Portuguese General Hospital.

    PubMed

    Barbosa, Sofia; Sequeira, Márcia; Castro, Sara; Manso, Rita; Klut Câmara, Catarina; Trancas, Bruno; Borja-Santos, Nuno; Maia, Teresa

    2016-08-01

    Introdução: Os doentes afectos de patologia psiquiátrica apresentam maior risco de morte, tanto por causas naturais como não naturais. Este estudo avalia as causas de morte de todos os doentes de uma unidade de internamento de agudos de Psiquiatria num hospital geral em Portugal, ao longo de dezasseis anos (de 1998 a 2013). Material e Métodos: Vinte e um doentes morreram na unidade de internamento de doentes agudos entre 1998 e 2013 (média 1,3 por ano). As características demográficas, os diagnósticos médicos e psiquiátricos foram recolhidos através de um estudo retrospectivo que consistiu na análise dos processos clínicos da amostra selecionada. Os doentes transferidos para outras enfermarias durante o internamento não foram incluídos no estudo. Resultados: As doenças do sistema circulatório foram as causas de morte mais prevalentes, ocorrendo em 2/3 dos doentes, incluindo embolismo pulmonar (n = 6), acidente vascular cerebral (n = 3), arritmia cardíaca (n = 2), enfarte agudo do miocárdio (n = 1), rutura de aneurisma da aorta abdominal (n = 1) e insuficiência cardíaca (n = 1). Dois doentes morreram de pneumonia e em quatro casos a causa de morte foi indeterminada. Apenas um caso de suicídio foi registado. Discussão: As doenças do aparelho circulatório foram as causas de morte mais frequentes nesta unidade de agudos. O suicídio em doentes internados, apesar de constituir um evento raro, é uma realidade que comporta consequências complexas para os profissionais de saúde, familiares e restantes doentes, devendo permanecer como foco de prevenção continuada. Conclusão: Os estudos de mortalidade são importantes para determinar a qualidade dos cuidados de saúde e criar recomendações para medidas preventivas.

  17. Texas Hold 'em Online Poker: A Further Examination

    ERIC Educational Resources Information Center

    Hopley, Anthony A. B.; Dempsey, Kevin; Nicki, Richard

    2012-01-01

    Playing Texas Hold 'em Online Poker (THOP) is on the rise. However, there is relatively little research examining factors that contribute to problem gambling in poker players. The aim of this study was to extend the research findings of Hopley and Nicki (2010). The negative mood states of depression, anxiety and stress were found to be linked to…

  18. Texas Hold 'em Online Poker: A Further Ex