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Sample records for avium isolates originating

  1. Relatedness of Mycobacterium avium subspecies hominissuis clinical isolates of human and porcine origins assessed by MLVA.

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    Leão, Célia; Canto, Ana; Machado, Diana; Sanches, Ilda Santos; Couto, Isabel; Viveiros, Miguel; Inácio, João; Botelho, Ana

    2014-09-17

    Mycobacterium avium subsp. hominissuis (MAH) is an important opportunistic pathogen, infecting humans and animals, notably pigs. Several methods have been used to characterize MAH strains. RFLP and PFGE typing techniques have been used as standard methods but are technically demanding. In contrast, the analysis of VNTR loci is a simpler, affordable and highly reliable PCR-based technique, allowing a numerical and reproductive digitalization of typing data. In this study, the analysis of Mycobacterium avium tandem repeats (MATRs) loci was adapted to evaluate the genetic diversity of epidemiological unrelated MAH clinical strains of human (n=28) and porcine (n=69) origins, collected from diverse geographical regions across mainland Portugal. These MAH isolates were found to be genetically diverse and genotypes are randomly distributed across the country. Some of the human strains shared identical VNTR profiles with porcine isolates. Our study shows that the VNTR genotyping using selected MATR loci is a useful analysis technique for assessing the genetic diversity of MAH isolates from Portugal. This typing method could be successfully applied in other countries toward the implementation of a worldwide open-access database of MATR-VNTR profiles of MAH isolates, allowing a better assessment of the global epidemiology traits of this important pathogenic species.

  2. Application of IS1311 locus 2 PCR-REA assay for the specific detection of ′Bison type′ Mycobacterium avium subspecies paratuberculosis isolates of Indian origin

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    Ajay Vir Singh

    2015-01-01

    Full Text Available Background & objectives: Of the three major genotypes of Mycobacterium avium subspecies paratuberculosis (MAP, ′Bison type′ is most prevalent genotype in the domestic livestock species of the country, and has also been recovered from patients suffering from Crohn′s disease. Recently, a new assay based on IS1311 locus 2 PCR- restriction endonuclease analysis (REA was designed to distinguish between ′Indian Bison type′ and non-Indian genotypes. The present study investigated discriminatory potential of this new assay while screening of a panel of MAP isolates of diverse genotypes and from different geographical regions. Methods: A total of 53 mycobacterial isolates (41 MAP and 12 mycobacterium other than MAP, three MAP genomic DNA and 36 MAP positive faecal DNA samples from different livestock species (cattle, buffaloes, goat, sheep and bison and geographical regions (India, Canada, USA, Spain and Portugal were included in the study. The extracted DNA samples (n=92 were analyzed for the presence of MAP specific sequences (IS900, ISMav 2 and HspX using PCR. DNA samples were further subjected to genotype differentiation using IS1311 PCR-REA and IS1311 L2 PCR-REA methods. Results: All the DNA samples (except DNA from non-MAP mycobacterial isolates were positive for all the three MAP specific sequences based PCRs. IS1311 PCR-REA showed that MAP DNA samples of Indian origin belonged to ′Bison type′. Whereas, of the total 19 non-Indian MAP DNA samples, 2, 15 and 2 were genotyped as ′Bison type′, ′Cattle type′ and ′Sheep type′, respectively. IS1311 L2 PCR-REA method showed different restriction profiles of ′Bison type′ genotype as compared to non-Indian DNA samples. Interpretation & conclusions: IS1311 L2 PCR-REA method successfully discriminated ′Indian Bison type′ from other non-Indian genotypes and showed potential to be future epidemiological tool and for genotyping of MAP isolates.

  3. Genotyping of Mycobacterium avium subsp. avium isolates from naturally infected lofts of domestic pigeons in Ahvaz by IS901 RFLP

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    Kaveh Parvandar Asadollahi

    2015-12-01

    Full Text Available Background and Objectives: Avian tuberculosis is one of the most important infections affecting most species of birds.Mycobacterium avium can not only infect all species of birds, but also infect some domesticated mammals.The most crucial aspect of control and eradication scheme is identification of infection sources and transmission routs. Mo- lecular techniques such as restriction fragment length polymorphism and pulse field gel electrophoresis have been shown to be much more discriminatory and suitable for use in the epidemiological study.Materials and Methods: Eighty suspected pigeons to avian tuberculosis based on their clinical signs, were subjected to the study. Forty Mycobacterium avium subsp. avium isolates out of a total of 51 identified isolates were subjected to the test.Results: IS901-RFLP using Pvu II was successfully conducted and produced 7 patterns. The majority of isolates (60% were RFLP type PI.1. This type was the most similar type to standard strain. However, all the patterns obtained in this study were different from the standard strain.Conclusion: The result of this study indicate that these isolates probably are limited to Khuzestan region. We recommend DNA fingerprinting differentiation of non tuberculous Mycobacteria particularly Mycobacterium avium complex isolated from infected birds and human to possibly find source of infections. Keywords: Mycobacterium avium, RFLP, polymorphism, avian tuberculosis

  4. Characterization of clinical and environmental Mycobacterium avium spp. isolates and their interaction with human macrophages.

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    Evelyn Guirado

    Full Text Available Members of the Mycobacterium avium complex (MAC are naturally occurring bacteria in the environment. A link has been suggested between M. avium strains in drinking water and clinical isolates from infected individuals. There is a need to develop new screening methodologies that can identify specific virulence properties of M. avium isolates found in water that predict a level of risk to exposed individuals. In this work we have characterized 15 clinical and environmental M. avium spp. isolates provided by the US Environmental Protection Agency (EPA to improve our understanding of the key processes involved in the binding, uptake and survival of these isolates in primary human macrophages. M. avium serovar 8 was predominant among the isolates studied. Different amounts and exposure of mannose-capped lipoarabinomannan (ManLAM and glycopeptidolipids (GPLs, both major mycobacterial virulence factors, were found among the isolates studied. Reference clinical isolate 104 serovar 1 and clinical isolates 11 and 14 serovar 8 showed an increased association with macrophages. Serum opsonization increased the cell association and survival at 2 h post infection for all isolates. However, only the clinical isolates 104 and 3 among those tested showed an increased growth in primary human macrophages. The other isolates varied in their survival in these cells. Thus we conclude that the amounts of cell envelope ManLAM and GPL, as well as GPL serovar specificity are not the only important bacterial factors for dictating the early interactions of M. avium with human macrophages.

  5. Host responses to persistent Mycobacterium avium subspecies paratuberculosis infection in surgically isolated bovine ileal segments.

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    Charavaryamath, Chandrashekhar; Gonzalez-Cano, Patricia; Fries, Patrick; Gomis, Susantha; Doig, Kimberley; Scruten, Erin; Potter, Andrew; Napper, Scott; Griebel, Philip J

    2013-02-01

    A lack of appropriate disease models has limited our understanding of the pathogenesis of persistent enteric infections with Mycobacterium avium subsp. paratuberculosis. A model was developed for the controlled delivery of a defined dose of M. avium subsp. paratuberculosis to surgically isolated ileal segments in newborn calves. The stable intestinal segments enabled the characterization of host responses to persistent M. avium subsp. paratuberculosis infections after a 9-month period, including an analysis of local mucosal immune responses relative to an adjacent uninfected intestinal compartment. M. avium subsp. paratuberculosis remained localized at the initial site of intestinal infection and was not detected by PCR in the mesenteric lymph node. M. avium subsp. paratuberculosis-specific T cell proliferative responses included both CD4 and γδ T cell receptor (γδTcR) T cell responses in the draining mesenteric lymph node. The levels of CD8(+) and γδTcR(+) T cells increased significantly (P avium subsp. paratuberculosis-specific tumor necrosis factor alpha (TNF-α) and gamma interferon secretion by lamina propria leukocytes was also significantly (P avium subsp. paratuberculosis infection. In conclusion, surgically isolated ileal segments provided a model system for the establishment of a persistent and localized enteric M. avium subsp. paratuberculosis infection in cattle and facilitated the analysis of M. avium subsp. paratuberculosis-specific changes in mucosal leukocyte phenotype and function. The accumulation of DC subpopulations in the lamina propria suggests that further investigation of mucosal DCs may provide insight into host responses to M. avium subsp. paratuberculosis infection and improve vaccine strategies to prevent M. avium subsp. paratuberculosis infection.

  6. Mycobacterium Avium subsp. paratuberculosis isolates induce in vitro granuloma formation and show successful survival phenotype, common anti-inflammatory and antiapoptotic responses within ovine macrophages regardless of genotype or host of origin.

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    Naiara Abendaño

    Full Text Available The analysis of the early macrophage responses, including bacterial growth within macrophages, represents a powerful tool to characterize the virulence of clinical isolates of Mycobcaterium avium susbp. paratuberculosis (Map. The present study represents the first assessment of the intracellular behaviour in ovine monocyte-derived macrophages (MDMs of Map isolates representing distinct genotypes (C, S and B, and isolated from cattle, sheep, goat, fallow deer, deer, and wild boar. Intracellular growth and survival of the selected isolates in ovine MDMs was assessed by quantification of CFUs inside of the host cells at 2 h p.i. (day 0 and 7 d p. i. using an automatic liquid culture system (Bactec MGIT 960. Variations in bacterial counts over 7 days from the baseline were small, in a range between 1.63 to 1.05-fold. After 7 d of infection, variations in the estimated log10 CFUs between all the tested isolates were not statistically significant. In addition, ovine MDMs exhibited enhanced anti-inflammatory, antiapoptotic and antidestructive responses when infected with two ovine isolates of distinct genotype (C and S or with two C-type isolates from distinct hosts (cattle and sheep; which correlated with the successful survival of these isolates within ovine MDMs. A second objective was to study, based on an in vitro granuloma model, latter stages of the infection by investigating the capacity of two Map isolates from cattle and sheep to trigger formation of microgranulomas. Upon 10 d p.i., both Map isolates were able to induce the formation of granulomas comparable to the granulomas observed in clinical specimens with respect to the cellular components involved. In summary, our results demonstrated that Map isolates from cattle, sheep, goats, deer, fallow-deer and wild boar were able not only to initiate but also to establish a successful infection in ovine macrophages regardless of genotype.

  7. Mycobacterium Avium subsp. paratuberculosis isolates induce in vitro granuloma formation and show successful survival phenotype, common anti-inflammatory and antiapoptotic responses within ovine macrophages regardless of genotype or host of origin.

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    Abendaño, Naiara; Tyukalova, Lyudmila; Barandika, Jesse F; Balseiro, Ana; Sevilla, Iker A; Garrido, Joseba M; Juste, Ramon A; Alonso-Hearn, Marta

    2014-01-01

    The analysis of the early macrophage responses, including bacterial growth within macrophages, represents a powerful tool to characterize the virulence of clinical isolates of Mycobcaterium avium susbp. paratuberculosis (Map). The present study represents the first assessment of the intracellular behaviour in ovine monocyte-derived macrophages (MDMs) of Map isolates representing distinct genotypes (C, S and B), and isolated from cattle, sheep, goat, fallow deer, deer, and wild boar. Intracellular growth and survival of the selected isolates in ovine MDMs was assessed by quantification of CFUs inside of the host cells at 2 h p.i. (day 0) and 7 d p. i. using an automatic liquid culture system (Bactec MGIT 960). Variations in bacterial counts over 7 days from the baseline were small, in a range between 1.63 to 1.05-fold. After 7 d of infection, variations in the estimated log10 CFUs between all the tested isolates were not statistically significant. In addition, ovine MDMs exhibited enhanced anti-inflammatory, antiapoptotic and antidestructive responses when infected with two ovine isolates of distinct genotype (C and S) or with two C-type isolates from distinct hosts (cattle and sheep); which correlated with the successful survival of these isolates within ovine MDMs. A second objective was to study, based on an in vitro granuloma model, latter stages of the infection by investigating the capacity of two Map isolates from cattle and sheep to trigger formation of microgranulomas. Upon 10 d p.i., both Map isolates were able to induce the formation of granulomas comparable to the granulomas observed in clinical specimens with respect to the cellular components involved. In summary, our results demonstrated that Map isolates from cattle, sheep, goats, deer, fallow-deer and wild boar were able not only to initiate but also to establish a successful infection in ovine macrophages regardless of genotype.

  8. First report of MIRU-VNTR genotyping of Mycobacterium avium subsp. paratuberculosis isolates from Egypt

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    A. Fawzy; Fayed, A.; Youssef, H; El-Sayed, A.; Zschöck, M.

    2016-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease, an economically important disease in ruminants worldwide. It was first isolated in Egypt in 2005. Since then, the pathogen has been detected in different Egyptian provinces. In order to trace the source of infection, genotyping using simple methods of high discriminatory power such as mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTR) were carried out in different co...

  9. Comparative transcriptional analysis of human macrophages exposed to animal and human isolates of Mycobacterium avium subspecies paratuberculosis with diverse genotypes.

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    Motiwala, Alifiya S; Janagama, Harish K; Paustian, Michael L; Zhu, Xiaochun; Bannantine, John P; Kapur, Vivek; Sreevatsan, Srinand

    2006-11-01

    Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease in animals and has been hypothesized to be associated with Crohn's disease in humans. Recently, M. avium subsp. paratuberculosis isolates recovered from Crohn's disease patients were shown to have limited diversity, implying the existence of human disease-associated genotypes and strain sharing with animals (A. H. Ghadiali et al., J. Clin. Microbiol. 42:5345-5348, 2004). To explore whether these genotypic differences or similarities among human and animal isolates translated to functionally significant attributes such as variance in host preference and/or difference in magnitude of infections, we performed a global scale analysis of M. avium subsp. paratuberculosis isolates that were representative of different genotypes and host species using DNA microarrays. Genome-wide characterization of the transcriptional changes was carried out using a human monocytic cell line (THP-1 cells) in response to different genotypes of M. avium subsp. paratuberculosis isolates recovered from various hosts. We identified several differentially expressed genes during early intracellular infection, including those involved in common canonical pathways such as NF-kappaB, interleukin-6 (IL-6), mitogen-activated protein kinase/extracellular signal-regulated kinase, and Jun N-terminal protein kinase signaling, as well as genes involved in T helper type 1 (Th1) responses (such as CCL5 ligand) and those that encode several proinflammatory cytokines and chemokine receptors. The cattle and human isolates of M. avium subsp. paratuberculosis, regardless of their short sequence repeat (SSR) genotype, induced similar global gene expression patterns in THP-1 cells. They differentially regulated genes necessary for cell survival without causing major alterations in proinflammatory genes. In contrast, the sheep isolates representing diverse SSR genotypes closely resembled the global gene expression pattern of an M

  10. MIRU-VNTR genotype diversity and indications of homoplasy in M. avium strains isolated from humans and slaughter pigs in Latvia.

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    Kalvisa, Adrija; Tsirogiannis, Constantinos; Silamikelis, Ivars; Skenders, Girts; Broka, Lonija; Zirnitis, Agris; Jansone, Inta; Ranka, Renate

    2016-09-01

    Diseases which are caused by non-tuberculous mycobacteria (NTM) are an increasing problem in the developed countries. In Latvia, one of the most clinically important members of NTM is Mycobacterium avium (M. avium), an opportunistic pathogen which has been isolated from several lung disease patients and tissue samples of slaughter pigs. This study was designed to characterize the genetic diversity of the M. avium isolates in Latvia and to compare the distribution of genotypic patterns among humans and pigs. Eleven (Hall and Salipante, 2010) clinical M. avium samples, isolated from patients of Center of Tuberculosis and Lung Diseases (years 2003-2010), and 32 isolates from pig necrotic mesenterial lymph nodes in different regions (years 2003-2007) were analyzed. The majority (42 of 43) of samples were identified as M. avium subsp. hominissuis; one porcine isolate belonged to M. avium subsp. avium. MIRU-VNTR genotyping revealed 13 distinct genotypes, among which nine genotype patterns, including M. avium subsp. avium isolate, were newly identified. IS1245 RFLP fingerprinting of 25 M. avium subsp. hominissuis samples yielded 17 different IS1245 RFLP patterns, allowing an efficient discrimination of isolates. Clusters of identical RFLP profiles were observed within host species, geographical locations and time frame of several years. Additional in silico analysis on simulated MIRU-VNTR genotype population datasets showed that the MIRU-VNTR pattern similarity could partly arise due to probabilistic increase of acquiring homoplasy among subpopulations, thus the similar MIRU-VNTR profiles of M. avium strains even in close geographical proximity should be interpreted with caution.

  11. Typing of Human Mycobacterium avium Isolates in Italy by IS1245-Based Restriction Fragment Length Polymorphism Analysis

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    Lari, Nicoletta; Cavallini, Michela; Rindi, Laura; Iona, Elisabetta; Fattorini, Lanfranco; Garzelli, Carlo

    1998-01-01

    All but 2 of 63 Mycobacterium avium isolates from distinct geographic areas of Italy exhibited markedly polymorphic, multibanded IS1245 restriction fragment length polymorphism (RFLP) patterns; 2 isolates showed the low-number banding pattern typical of bird isolates. By computer analysis, 41 distinct IS1245 patterns and 10 clusters of essentially identical strains were detected; 40% of the 63 isolates showed genetic relatedness, suggesting the existence of a predominant AIDS-associated IS1245 RFLP pattern. PMID:9817900

  12. Distribution of serotypes, IS901 and a 40 kDa protein in Mycobacterium avium complex strains isolated from man and animals in Denmark

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    Klausen, J.; Giese, Steen Bjørck; Fuursted, K.;

    1997-01-01

    The aim of the present study was to characterize all strains of the Mycobacterium avium complex isolated in Denmark in 1993. A total of 141 M. avium complex strains (86 from man, 38 from animals, and 17 from peat) were analysed by serotyping, ELISA specific for a 40 kDa protein, and IS901-specific...... prevalent serotypes among strains isolated from peat were serotype 4 (29%) and serotype 9 (18%). There was a concurrent appearance of IS901 and p40 in all strains. Only M. avium complex strains isolated from animals, and belonging to serotype 1 or serotype 2, contained the IS901/p40 markers. The different...

  13. Isolation and characterization of a novel bacteriophage against Mycobacterium avium subspecies paratuberculosis.

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    Basra, Simone; Anany, Hany; Brovko, Lioubov; Kropinski, Andrew M; Griffiths, Mansel W

    2014-10-01

    Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne's disease, has a doubling time of 24 hours, making rapid detection very difficult. Mycobacteriophages can be used in the detection of disease-causing mycobacteria such as MAP. Isolation and sequencing the genomes of lytic MAP bacteriophages are important preliminary steps towards designing phage-based rapid detection assays for this bacterium. A simple optimized protocol was developed to allow reproducible production of confluent growth of MAP on plates within four to six weeks of incubation at 30 °C. This protocol was applied to the screening of environmental and fecal samples for bacteriophages inhibiting the growth of MAP. As a result, a lytic phage, vB_MapS_FF47, was isolated from bovine feces. FF47 contains a double-stranded DNA genome ~48 kb in length with 73 protein coding sequences. It does not carry temperate or known virulence genes. This phage was shown to be most closely related to Mycobacterium phage Muddy, isolated in South Africa, and Gordonia phage GTE2; however, it could not infect any of the tested Gordonia, Rhodococcus, or Nocardia spp. that GTE2 could. The protocols that were developed for growth and phage isolation have potential applications in a high-throughput screening for compounds inhibiting the growth of MAP. This work describes the first time that a phage was isolated against M. paratuberculosis.

  14. Full genome sequence of a Danish isolate of Mycobacterium avium subspecies paratuberculosis, strain Ejlskov2007

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    Afzal, Mamuna; Abidi, Soad; Mikkelsen, Heidi

    , consisting of 4317 unique gene families. Comparison with M. avium paratuberculosis strain K10 revealed only 3436 genes in common (~70%). We have used GenomeAtlases to show conserved (and unique) regions along the Ejlskov2007 chromosome, compared to 2 other Mycobacterium avium sequenced genomes. Pan-genome...

  15. First report of MIRU-VNTR genotyping of Mycobacterium avium subsp. paratuberculosis isolates from Egypt.

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    Fawzy, A; Fayed, A; Youssef, H; El-Sayed, A; Zschöck, M

    2016-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne's disease, an economically important disease in ruminants worldwide. It was first isolated in Egypt in 2005. Since then, the pathogen has been detected in different Egyptian provinces. In order to trace the source of infection, genotyping using simple methods of high discriminatory power such as mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTR) were carried out in different countries. Until now there is no published information about MIRU-VNTR genotyping of MAP isolates in Egypt. To address that point, 100 faecal samples were collected and cultivated from 3 different suspected dairy farms. Fourteen isolates belonging to one farm were identified as MAP and subjected to genotyping using 8 different MIRU-VNTR loci PCRs. Two different genotypes were recognized based on size polymorphism observed in one locus (VNTR-7) that was confirmed by sequencing. Our work provides a preliminary basis of constructing a MIRU-VNTR genotyping database of MAP in Egypt.

  16. Isolation of Mycobacterium avium complex from bone marrow aspirates of AIDS patients in Brazil.

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    Barreto, J A; Palaci, M; Ferrazoli, L; Martins, M C; Suleiman, J; Lorenço, R; Ferreira, O C; Riley, L W; Johnson, W D; Galvão, P A

    1993-09-01

    Mycobacterium avium complex (MAC) infection has not been reported as a major opportunistic infection among patients with AIDS in Latin America or Africa. In this study, 125 AIDS patients who had persistent fever, anemia, and leukopenia were examined among 2628 AIDS patients admitted to Instituto de Infectologia Emilio Ribas between May 1990 and April 1992. From the bone marrow aspirates of the 125 patients, MAC was isolated from 23 (18.4%) and Mycobacterium tuberculosis was isolated from 9 (7.2%). Between 1985 and 1990, only 11 MAC isolations among 60,000 cultures obtained from human immunodeficiency virus-seronegative patients were documented in São Paulo. Hence, the minimal estimated rate of MAC infection in AIDS patients in this city was 23/2628, or 0.88%. These findings suggest that MAC infection is an important opportunistic infection, especially among a subset of patients with AIDS in Brazil who have clinical characteristics and risk activities similar to those associated with MAC infections in North America and Europe.

  17. Genetic diversity of Mycobacterium avium isolates recovered from clinical samples and from the environment: molecular characterization for diagnostic purposes.

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    Alvarez, Julio; García, Ignacio Gómez; Aranaz, Alicia; Bezos, Javier; Romero, Beatriz; de Juan, Lucía; Mateos, Ana; Gómez-Mampaso, Enrique; Domínguez, Lucas

    2008-04-01

    Isolation of Mycobacterium avium complex (MAC) organisms from clinical samples may occur in patients without clinical disease, making the interpretation of results difficult. The clinical relevance of MAC isolates from different types of clinical samples (n = 47) from 39 patients in different sections of a hospital was assessed by comparison with environmental isolates (n = 17) from the hospital. Various methods for identification and typing (commercial probes, phenotypic characteristics, PCR for detection of IS1245 and IS901, sequencing of the hsp65 gene, and pulsed-field gel electrophoresis) were evaluated. The same strain was found in all the environmental isolates, 21 out of 23 (91.3%) of the isolates cultured from urine samples, and 5 out of 19 (26.3%) isolates from respiratory specimens. This strain did not cause disease in the patients. Testing best characterized the strain as M. avium subsp. hominissuis, with the unusual feature that 81.4% of these isolates lacked the IS1245 element. Contamination of certain clinical samples with an environmental strain was the most likely event; therefore, characterization of the environmental mycobacteria present in health care facilities should be performed to discard false-positive isolations in nonsterile samples, mainly urine samples. Molecular techniques applied in this study demonstrated their usefulness for this purpose.

  18. AMPLIFIED FRAGMENT LENGTH POLYMORPHISM ANALYSIS OF MYCOBACTERIUM AVIUM COMPLEX ISOLATES RECOVERED FROM SOUTHERN CALIFORNIA

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    Fine-scale genotyping methods are necessary in order to identify possible sources of human exposure to opportunistic pathogens belonging to the Mycobacterium avium complex (MAC). In this study, amplified fragment length polymorphism (AFLP) analysis was evaluated for fingerprintin...

  19. Isolation of flavonoids from the heartwood and resin of Prunus avium and some preliminary biological investigations.

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    McNulty, James; Nair, Jerald J; Bollareddy, Endreddy; Keskar, Kunal; Thorat, Amol; Crankshaw, Denis J; Holloway, Alison C; Khan, Ghaznia; Wright, Gerard D; Ejim, Linda

    2009-12-01

    An investigation of the constituents in heartwood and resin of Prunus avium is reported. A mini-library of structurally diverse flavanones and flavones was screened for human cytochrome P450 1A1, 3A4 and 19 (aromatase) inhibition, and for antifungal activity against a panel of pathogenic fungi. The defensive role of these natural plant flavonoids as antifungal phytoalexins and phytoanticipins is discussed.

  20. Differential proteome analysis of Mycobacterium avium subsp. paratuberculosis grown in vitro and isolated from cases of clinical Johne's disease.

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    Weigoldt, Mathias; Meens, Jochen; Doll, Klaus; Fritsch, Isabel; Möbius, Petra; Goethe, Ralph; Gerlach, Gerald F

    2011-02-01

    Bovine Johne's disease (paratuberculosis), caused by Mycobacterium avium subspecies paratuberculosis, poses a significant economic problem to the beef and dairy industry worldwide. Despite its relevance, however, pathogenesis of Johne's disease is still only partially resolved. Since mycobacterial membrane proteins expressed during infection are likely to play an important role in pathogenesis, membrane-enriched fractions, namely mucosa-derived membranes (MDM) and culture-derived membranes (CDM), of M. avium subsp. paratuberculosis from three cows with clinical paratuberculosis were investigated. An initial analysis by 2D difference gel electrophoresis (2D DIGE) and MALDI-TOF-MS analysis revealed four differentially expressed proteins with only one predicted membrane protein. Due to this limited outcome, membrane preparations were subjected to a tube-gel trypsin digestion and investigated by using nanoflow-liquid-chromatography-coupled tandem MS. Based on this approach a total of 212 proteins were detected in MDM including 32 proteins of bovine origin; 275 proteins were detected in CDM; 59 % of MDM and CDM proteins were predicted to be membrane-associated. A total of 130 of the proteins were detected in both MDM and CDM and 48 predicted membrane proteins were detected in MDM from at least two cows. Four of these proteins were not detected in CDM, implying differential expression in the host. All membrane-associated proteins, especially the four identified as being differentially expressed, might be relevant targets for further analyses into the pathogenesis of bovine paratuberculosis.

  1. First isolation of mycobacterium avium subsp. Paratuberculosis from wild guanacos (Lama guanicoe) on Tierra del Fuego Island.

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    Salgado, M; Herthnek, D; Bölske, G; Leiva, S; Kruze, J

    2009-04-01

    The aim of this study was to search for Mycobacterium avium subsp. paratuberculosis (Map) infection in a free-ranging wild animal species in a region where Johnes's disease has yet to be reported and to classify Map isolates using a genomic typing method. Fecal samples were obtained from 501 wild guanacos (Lama guanicoe) from Tierra del Fuego Island, Chile, in August 2006. Samples were cultured using Herrold's egg yolk medium with and without mycobactin J. After 9 mo of incubation, suspected Map colonies showing mycobactin dependence were confirmed by real-time polymerase chain reaction (PCR) based on IS900 and F57. Isolates were further tested using IS1311 PCR with restriction endonuclease analysis in order to type the guanaco Map strains. Twenty-one of 501 (4.2%) animals were fecal culture-positive for Map; identity was confirmed by real-time PCR and isolates were classified as cattle-type. Most culture-positive animals were located in four contiguous geographic areas, and the infection was most commonly found among adult animals. Prevalence was higher in females (5.9%) than males (3.1%) but the difference was not statistically significant. This represents the first isolation of Map from a free-ranging wildlife species in Chile. It expands the geographic range of paratuberculosis and the diversity of wildlife species that can become infected with Map.

  2. Rapid susceptibility testing of Mycobacterium avium complex and Mycobacterium tuberculosis isolated from AIDS patients

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    Dhople, Arvind M.

    1994-01-01

    In ominous projections issued by both U.S. Public Health Service and the World Health Organization, the epidemic of HIV infection will continue to rise more rapidly worldwide than predicted earlier. The AIDS patients are susceptible to diseases called opportunistic infections of which tuberculosis and Mycobacterium avium complex (MAC) infection are most common. This has created an urgent need to uncover new drugs for the treatment of these infections. In the seventies, NASA scientists at Goddard Space Flight Center, Greenbelt, MD, had adopted a biochemical indicator, adenosine triphosphate (ATP), to detect presence of life in extraterrestrial space. We proposed to develop ATP assay technique to determine sensitivity of antibacterial compounds against MAC and M. tuberculosis.

  3. Isolated Endobronchial Mycobacterium avium Disease Associated with Lobar Atelectasis in an Immunocompetent Young Adult: A Case Report and Literature Review.

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    Kim, Hye In; Kim, Ji Won; Kim, Jun Young; Kim, Young Nam; Kim, Jin Hae; Jeong, Byeong-Ho; Chung, Myung Jin; Koh, Won-Jung

    2015-10-01

    The prevalence of lung diseases caused by nontuberculous mycobacteria (NTM) is increasing worldwide. Unlike pulmonary tuberculosis, endobronchial NTM diseases are very rare with the majority of cases reported in patients with human immunodeficiency virus infection and acquired immune deficiency syndrome. We reported a rare case of endobronchial Mycobacterium avium disease associated with lobar atelectasis in a young immunocompetent patient and reviewed the relevant iterature.

  4. Isolation of Mycobacterium avium subspecies paratuberculosis Reactive T-cells from Intestinal Biopsies of Crohn's Disease Patients

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    Crohn’s disease (CD) is a chronic granulomatous inflammation of the intestine. The etiology is still unknown. One hypothesis is that CD is caused by infection with Mycobacterium avium subspecies paratuberculosis (MAP) in genetically predisposed individuals. MAP causes a similar disease in ruminants,...

  5. Genome-wide sequence variations among Mycobacterium avium subspecies paratuberculosis.

    Directory of Open Access Journals (Sweden)

    Chung-Yi eHsu

    2011-12-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (M. ap, the causative agent of Johne’s disease (JD, infects many farmed ruminants, wildlife animals and humans. To better understand the molecular pathogenesis of these infections, we analyzed the whole genome sequences of several M. ap and M. avium subspecies avium (M. avium strains isolated from various hosts and environments. Using Next-generation sequencing technology, all 6 M. ap isolates showed a high percentage of homology (98% to the reference genome sequence of M. ap K-10 isolated from cattle. However, 2 M. avium isolates (DT 78 and Env 77 showed significant sequence diversity from the reference strain M. avium 104. The genomes of M. avium isolates DT 78 and Env 77 exhibited only 87% and 40% homology, respectively, to the M. avium 104 reference genome. Within the M. ap isolates, genomic rearrangements (insertions/deletions, Indels were not detected, and only unique single nucleotide polymorphisms (SNPs were observed among the 6 M. ap strains. While most of the SNPs (~100 in M. ap genomes were non-synonymous, a total of ~ 6000 SNPs were detected among M. avium genomes, most of them were synonymous suggesting a differential selective pressure between M. ap and M. avium isolates. In addition, SNPs-based phylo-genomic analysis showed that isolates from goat and Oryx are closely related to the cattle (K-10 strain while the human isolate (M. ap 4B is closely related to the environmental strains, indicating environmental source to human infections. Overall, SNPs were the most common variations among M. ap isolates while SNPs in addition to Indels were prevalent among M. avium isolates. Genomic variations will be useful in designing host-specific markers for the analysis of mycobacterial evolution and for developing novel diagnostics directed against Johne’s disease in animals.

  6. Mycobacterium avium subspecies paratuberculosis isolates from sheep and goats show reduced persistence in bovine macrophages than cattle, bison, deer and wild boar strains regardless of genotype.

    Science.gov (United States)

    Abendaño, Naiara; Sevilla, Iker A; Prieto, José Miguel; Garrido, Joseba M; Juste, Ramon A; Alonso-Hearn, Marta

    2013-05-03

    Assessment of the virulence of isolates of Mycobacterium avium subsp. paratuberculosis (Map) exhibiting distinct genotypes and isolated from different hosts may help to clarify the degree to which clinical manifestations of the disease in cattle can be attributed to bacterial or to host factors. The objective of this study was to test the ability of 10 isolates of Map representing distinct genotypes and isolated from domestic (cattle, sheep, and goat), and wildlife animal species (fallow deer, deer, wild boar, and bison) to enter and grow in bovine macrophages. The isolates were previously typed using IS1311 PCR followed by restriction endonuclease analysis into types C, S or B. Intracellular growth of the isolates in a bovine macrophage-like cell line (BoMac) and in primary bovine monocyte-derived macrophages (MDM) was evaluated by quantification of CFU numbers in the initial inoculum and inside of the host cells at 2h and 7 d p.i. using an automatic liquid culture system (Bactec MGIT 960). Individual data illustrated that growth was less variable in BoMac than in MDM cells. All the isolates from goat and sheep hosts persisted within BoMac cells in lower CFU numbers than the other tested isolates after 7 days of infection regardless of genotype. In addition, BoMac cells exhibited differential inflammatory, apoptotic and destructive responses when infected with a bovine or an ovine isolate; which correlated with the differential survival of these strains within BoMac cells. Our results indicated that the survival of the tested Map isolates within bovine macrophages is associated with the specific host from which the isolates were initially isolated.

  7. XML Genetic Structure of SSR1 & SSR2 loci from Iranian Mycobacterium Avium Subspecies Paratuberculosis Isolates by a Short Sequence Repeat Analysis Approach

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    Aida Chalesh (MSc

    2016-01-01

    Full Text Available Background and Objective: Paratuberculosis has been repeatedly reported from Iranian ruminant herds. The extrem fastidious nature of Mycobacterium avium subspecies paratuberculsos hinders genomic diversity studies of the pathogen. Short Sequence Repeat analysis is one of the genome-based approches recently developed to overcome this difficulty. In this study we describe the application of SSR genotyping on three Iranian MAP type strains plus the III & V vaccinal strain. Methods: All the bacteria were examined by PCR-F57 and PCR-IS900 experiments in order to authenticate their identity as MAP. SSR genotyping using SSR1 & SSR2 loci was conducted according to the Amonsin method. PCR amplicons were sequenced to guarantee the accuracy of findings. Results: At SSR1 locus two allels were identified, a larger allel of 770 bp and a smaller allel of 763 bp long. At SSR2 only a single allele, 800 bp long, was detected. Two Iranian bovine and ovine MAP isolates along with the vaccinal III & V strain shared a single SSR1/SSR2 pattern while a different SSR1/SSR2 was represented by the third (caprine Iranian MAP isolate. Conclusion: While finding a shared SSR type between the two Iranian MAP isolates and the III & V strain might represent a mutual ancestral background but this has to be assessed through further studies. Detection of two SSR genotypes between three Iranian type strains is likely a reflection of more MAP clones in Iran.

  8. Chlorine, Chloramine, Chlorine Dioxide, and Ozone Susceptibility of Mycobacterium avium

    OpenAIRE

    Taylor, Robert H; Falkinham, Joseph O.; Norton, Cheryl D.; LeChevallier, Mark W.

    2000-01-01

    Environmental and patient isolates of Mycobacterium avium were resistant to chlorine, monochloramine, chlorine dioxide, and ozone. For chlorine, the product of the disinfectant concentration (in parts per million) and the time (in minutes) to 99.9% inactivation for five M. avium strains ranged from 51 to 204. Chlorine susceptibility of cells was the same in washed cultures containing aggregates and in reduced aggregate fractions lacking aggregates. Cells of the more slowly growing strains wer...

  9. Isolation and detection of Mycobacterium avium subsp. paratuberculosis (MAP from cattle in Ireland using both traditional culture and molecular based methods

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    Douarre Pierre E

    2010-09-01

    Full Text Available Abstract Background Mycobacterium avium subsp. paratuberculosis (MAP causes a chronic gastroenteritis affecting many species. Johne's disease is one of the most widespread and economically important disease of ruminants. Since 1992 and the opening of the European market, the exposure and the transmission of MAP in cattle herds considerably increased. Improvements in diagnostic strategies for Ireland and elsewhere are urgently required. In total, 290 cattle from seven Irish herds with either a history or a strong likelihood of paratuberculosis infection were selected by a veterinary team over 2 years. Faecal samples (290 were collected and screened for MAP by a conventional culture method and two PCR assays. In order to further evaluate the usefulness of molecular testing, a nested PCR was also assessed. Results M. paratuberculosis was isolated and cultured from 23 faecal samples (7.9% on solid medium. From a molecular perspective, 105 faecal samples (36% were PCR positive for MAP specific DNA. A complete correlation (100% was observed between the results of both molecular targets (IS900 and ISMAP02. Sensitivity was increased by ~10% with the inclusion of a nested PCR for ISMAP02 (29 further samples were positive. When culturing and PCR were retrospectively compared, every culture positive faecal sample also yielded a PCR positive result for both targets. Alternatively, however not every PCR positive sample (n = 105, 36% produced a corresponding culture isolate. Interestingly though when analysed collectively at the herd level, the correlation between culture and PCR results was 100% (ie every herd which recorded at least 1 early PCR +ve result later yielded culture positive samples within that herd. Conclusion PCR on bovine faecal samples is a fast reliable test and should be applied routinely when screening for MAP within herds suspected of paratuberculosis. Nested PCR increases the threshold limit of detection for MAP DNA by approximately 10

  10. Sero-prevalence of bovine Johne's disease in buffaloes and cattle population of North India using indigenous ELISA kit based on native Mycobacterium avium subspecies paratuberculosis 'Bison type' genotype of goat origin.

    Science.gov (United States)

    Singh, S V; Singh, A V; Singh, R; Sharma, S; Shukla, N; Misra, S; Singh, P K; Sohal, J S; Kumar, H; Patil, P K; Misra, P; Sandhu, K S

    2008-09-01

    Present pilot study is the first attempt in the country to estimate sero-prevalence of Bovine Johne's disease (BJD) by screening cattle and buffaloes representing large population belonging to farmer's and farm herds in the home tracts (Uttar Pradesh (UP) and Punjab) of Hariana cattle and Murrah buffaloes in North India. Indigenous and in-house plate ELISA kit (using protoplasmic antigen from native Mycobacterium avium subsp. paratuberculosis 'Bison type' strain of goat origin), originally developed for goats and sheep was standardized in bovines and used for screening. For this study, 33 villages of south and west UP were randomly selected and surveyed from 2001 to 2003. There were 7943 farmer's families having 38,251 livestock, including cattle, buffaloes, goats and sheep (per family 4.8% livestock). Numerically, buffaloes and cattle were 54.7% and 22.1%, respectively. Serum samples were collected from 726 animals (4.2% of 16, 981 livestock with 4375 farmer's families) located in 33 randomly surveyed villages. Serum samples (699), submitted to Epidemiology Department of Veterinary College (Punjab Agricultural University, Ludhiana), in the year 2004 by farmer's and organized farm herds (Buffaloes, 372, Cattle, 327), were screened by this ELISA kit. Soluble protoplasmic antigen was prepared from Map (S 5) 'Bison type' strain isolated from a terminally sick goat with Johne's disease. Of the total 1425 bovine (Buffaloes and cattle) serum samples screened using indigenous ELISA kit, sero-prevalence of Johne's disease was 29.0% (28.6% in buffalo and 29.8% in cattle) in Northern India. State-wise sero-prevalence was 31.9% and 23.3% in UP and Punjab, respectively. In UP, of the 601 randomly sampled buffaloes, sero-prevalence was 40.3% (16.6% in young and 40.9% adults) and 25.5% (10.5% in young and 26.3% adults) in south and west UP, respectively. Of the 125 cattle screened, sero-prevalence was 42.6% (nil in young and 44.4% adults) and 30.0% (nil in young and 30.6% adults

  11. Aislamiento de Mycobacterium avium subsp. Paratuberculosis de fecas en rebaños lecheros infectados mediante el Método de Cornell modificado Isolation of Mycobacterium avium subsp. Paratuberculosis from bovine feces of infected dairy herds by the Cornell’s Method modified

    Directory of Open Access Journals (Sweden)

    J. P SOTO

    2002-01-01

    Full Text Available Con la finalidad de aumentar la tasa de aislamiento de Mycobacterium avium subsp. paratuberculosis (Map, a partir de muestras de fecas bovinas, se evaluó un nuevo procedimiento de descontaminación y cultivo de muestras fecales en 250 animales clínicamente sanos, provenientes de 14 rebaños infectados del sur de Chile. Para la descontaminación de las muestras previo al cultivo se utilizó una solución al 0.9% de cloruro de hexadecilpiridinio (HPC y una solución antibiótica con amfotericina B, vancomicina y ácido nalidíxico. Para el aislamiento del agente se utilizó el medio de Herrold con yema de huevo y micobactina J adicionado de una solución antibiótica similar a la empleada para la descontaminación de las muestras. En el 16% (40 de las muestras analizadas fue posible aislar Map con un 7.6% de contaminación con hongos sólo a partir de la octava semana de incubación. La identidad de las cepas aisladas fue confirmada en el 100% de los casos mediante PCR, utilizando partidores específicos para este agente (P90 y P91. La alta tasa de aislamiento, la especificidad del medio de cultivo y la baja tasa de contaminación de los cultivos, durante el prolongado período de incubación, hacen de este procedimiento una buena alternativa de diagnóstico de Paratuberculosis bovinaIn order to improve the isolation rate of Mycobacterium avium subsp. paratuberculosis (Map from bovine feces a new bacteriological procedure for decontamination and cultivation of fecal samples was evaluated in 250 samples collected from asymptomatic animals in 14 infected dairy herds in southern Chile. Before culture all samples were treated with a decontaminant solution containing 0.9% hexadecylpiridinium chloride and an antibiotic solution containing amphotericin B, vancomycin and nalidixic acid. Herrold Egg Yolk Medium (HEYM and mycobactin J supplemented with the same antibiotics mentioned above was used for the isolation of the agent. Map was isolated from 40

  12. Mycobacterium avium subsp. hominissuis Infection in Swine Associated with Peat Used for Bedding

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    Tone Bjordal Johansen

    2014-01-01

    Full Text Available Mycobacterium avium subsp. hominissuis is an environmental bacterium causing opportunistic infections in swine, resulting in economic losses. Additionally, the zoonotic aspect of such infections is of concern. In the southeastern region of Norway in 2009 and 2010, an increase in condemnation of pig carcasses with tuberculous lesions was seen at the meat inspection. The use of peat as bedding in the herds was suspected to be a common factor, and a project examining pigs and environmental samples from the herds was initiated. Lesions detected at meat inspection in pigs originating from 15 herds were sampled. Environmental samples including peat from six of the herds and from three peat production facilities were additionally collected. Samples were analysed by culture and isolates genotyped by MLVA analysis. Mycobacterium avium subsp. hominissuis was detected in 35 out of 46 pigs, in 16 out of 20 samples of peat, and in one sample of sawdust. MLVA analysis demonstrated identical isolates from peat and pigs within the same farms. Polyclonal infection was demonstrated by analysis of multiple isolates from the same pig. To conclude, the increase in condemnation of porcine carcasses at slaughter due to mycobacteriosis seemed to be related to untreated peat used as bedding.

  13. Chlorine, Chloramine, Chlorine Dioxide, and Ozone Susceptibility of Mycobacterium avium

    Science.gov (United States)

    Taylor, Robert H.; Falkinham, Joseph O.; Norton, Cheryl D.; LeChevallier, Mark W.

    2000-01-01

    Environmental and patient isolates of Mycobacterium avium were resistant to chlorine, monochloramine, chlorine dioxide, and ozone. For chlorine, the product of the disinfectant concentration (in parts per million) and the time (in minutes) to 99.9% inactivation for five M. avium strains ranged from 51 to 204. Chlorine susceptibility of cells was the same in washed cultures containing aggregates and in reduced aggregate fractions lacking aggregates. Cells of the more slowly growing strains were more resistant to chlorine than were cells of the more rapidly growing strains. Water-grown cells were 10-fold more resistant than medium-grown cells. Disinfectant resistance may be one factor promoting the persistence of M. avium in drinking water. PMID:10742264

  14. Comparação de duas técnicas de isolamento do Mycobacterium avium subsp. paratuberculosis em amostras de fezes de ovinos com suspeita clínica de paratuberculose Comparison of two techniques of isolation of Mycobacterium avium subsp. paratuberculosis in faecal samples of ovine with clinical suspicion of paratuberculosis

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    Ana Cláudia Coelho

    2009-05-01

    Full Text Available A paratuberculose é uma enterite crônica granulomatosa causada por Mycobacterium avium subsp. paratuberculosis que afeta principalmente os ruminantes. A cultura de bactérias a partir de amostras de fezes e tecidos constitui um dos métodos mais eficazes de diagnóstico, sendo ainda o único método disponível para obtenção de isolamentos e estirpes de micobactérias. Contudo, este método apresenta baixa sensibilidade e requer meses de incubação antes do crescimento de colônias. Neste estudo, utilizou-se a cultura fecal como método de diagnóstico em ovinos de diferentes raças portuguesas, com sinais compatíveis com a doença. Fez-se ainda a comparação entre os meios de cultura Löwenstein Jensen® com micobactina® J e o de Middlebrook® 7H11 com OADC®, utilizados no isolamento da bactéria. As percentagens de isolamento em cada um os meios foram de 2,0% (6/300 para Löwenstein Jensen® com micobactina J e 1,0% (3/300 para Middlebrook® 7H11/OADC. As três amostras positivas no meio de Middlebrook® 7H11/OADC também foram positivas no meio de Löwenstein Jensen® com micobactina J e nenhuma foi somente positiva no meio de Middlebrook® 7H11/OADC. Os resultados deste estudo sugerem que o meio de Löwenstein-Jensen® com micobactina® J é mais efetivo para a obtenção de estirpes ovinas em Portugal.Paratuberculosis is a chronic enteric disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis. Culture of bacteria from faeces and tissues samples constitutes one of the most effective methods of confirming the diagnosis of para-tuberculosis and the only method available to obtain strains of mycobacteria. However, this method is less sensitive and requires months of incubation before colony growth occurs. In this study, culture method was used on sheep faeces to diagnose paratuber-culosis in animals with compatible signs of the disease. A comparison of two culture media used to isolation was also investigated

  15. Lymphadenitis in children is caused by Mycobacterium avium hominissuis and not related to ‘bird tuberculosis’

    Science.gov (United States)

    de Haas, P. E. W.; Lindeboom, J. A.; Kuijper, E. J.; van Soolingen, D.

    2008-01-01

    Mycobacterium avium is the most commonly encountered mycobacterium species among non-Mycobacterium tuberculosis complex (nontuberculous mycobacteria) isolates worldwide and frequently causes lymphadenitis in children. During a multi-centre study in The Netherlands that was performed to determine the optimal treatment for mycobacterial lymphadenitis, concern was expressed in the media about the possible role of birds as sources of these M. avium infections, referred to as ‘bird tuberculosis.’ To examine the involvement of birds in mycobacterial lymphadenitis, 34 M. avium isolates from lymphadenitis cases were subjected to IS1245 restriction fragment length polymorphism (RFLP) typing. This genotyping method enables the distinction of the subspecies M. avium subsp. hominissuis and the ‘bird-type’ M. avium spp. avium. Highly variable RFLP patterns were found among the lymphadenitis M. avium isolates, and all belonged to the M. avium hominissuis subspecies. A relation to pet birds in the etiology of mycobacterial lymphadenitis could not be established, and the source of the infections may be environmental. PMID:18320245

  16. Mycobacterium avium complex (MAC isolated from AIDS patients and the criteria required for its implication in disease Complexo Mycobacterium avium (MAC isolado de pacientes com AIDS e os critérios exigidos para sua implicação em doença

    Directory of Open Access Journals (Sweden)

    David Jamil Hadad

    1995-10-01

    Full Text Available Before the AIDS pandemia, the Mycobacterium avium complex (MAC was responsible in most cases for the pneumopathies that attack patients with basic chronic pulmonary diseases such as emphysema and chronic bronchitis36. In 1981, with the advent of the acquired immunodeficiency syndrome (AIDS, MAC started to represent one of the most frequent bacterial diseases among AIDS patients, with the disseminated form of the disease being the major clinical manifestation of the infection8. Between January 1989 and February 1991, the Section of Mycobacteria of the Adolfo Lutz Institute, São Paulo, isolated MAC from 103 patients by culturing different sterile and no-sterile processed specimens collected from 2304 patients seen at the AIDS Reference and Training Center and/or Emilio Ribas Infectology Institute. Disseminated disease was diagnosed in 29 of those patients on the basis of MAC isolation from blood and/or bone marrow aspirate. The other 74 patients were divided into categories highly (5, moderately (26 and little suggestive of disease (43 according to the criteria of DAVIDSON (198910. The various criteria for MAC isolation from sterile and non-sterile specimens are discussed.Anterior a pandemia de AIDS, o Complexo Mycobacterium avium (MAC era responsável pela maioria das vezes, por pneumopatias acometendo pacientes com doença pulmonar crônica de base como enfisema e bronquite crônica36. Em 1981, com o advento da síndrome de imunodeficiência adquirida (SIDA, o MAC passou a representar uma das doenças bacterianas mais frequentes em pacientes com esta síndrome, sendo a doença disseminada a principal forma de manifestação clínica da infecção8. Entre Janeiro de 1989 e Fevereiro de 1991, no Setor de Micobactérias do Instituto Adolfo Lutz em São Paulo, o MAC foi isolado de 103 pacientes a partir do cultivo de diferentes espécimes estéreis e não estéreis processados, coletados de 2.304 pacientes atendidos no Centro de Referência e

  17. Distribution of IS901 in strains of Mycobacterium avium complex from swine by using IS901-detecting primers that discriminate between M. avium and Mycobacterium intracellulare.

    Science.gov (United States)

    Nishimori, K; Eguchi, M; Nakaoka, Y; Onodera, Y; Ito, T; Tanaka, K

    1995-08-01

    The presence of the mycobacterial insertion sequence IS901 was studied by PCR with reference strains of Mycobacterium avium complex; 122 veterinary strains of mycobacteria, mainly M. avium complex, isolated from swine; and 15 clinical strains. Four kinds of DNA extraction methods for PCR were compared. Use of the commercial extraction matrix allowed for the faster and easier preparation of PCR-amplifiable DNA than use of NaOH heating extraction or sodium dodecyl sulfate extraction of pretreated mycobacteria. It also provided more effective protection than boiling extraction against the destruction of DNA. Four reference strains of serovars 1 to 3 possessed IS901. Nine reference strains of serovars 1, 4 to 6, 8 to 11, and 21 possessed only IS901 insertion sites. A novel PCR product was found in the other reference strains of serovars 7, 12 to 17, 19, and 20 and two clinical strains of serovar 15. It is suggested that the primers that amplified the insertion portion of IS901 divided the M. avium complex into M. avium, Mycobacterium intracellulare, and other mycobacteria. None of the 110 strains of M. avium complex isolated from swine possessed IS901. It is suggested that the absence of IS901 might be characteristic of swine-derived strains of M. avium complex.

  18. Isolation of S-locus F-box alleles in Prunus avium and their application in a novel method to determine self-incompatibility genotype.

    Science.gov (United States)

    Vaughan, S P; Russell, K; Sargent, D J; Tobutt, K R

    2006-03-01

    This study characterises a series of 12 S-locus haplotype-specific F-box protein genes (SFB) in cherry (Prunus avium) that are likely candidates for the pollen component of gametophytic self-incompatibility in this species. Primers were designed to amplify 12 SFB alleles,including the introns present in the 50 untranslated region;sequences representing the S-alleles S1, S2, S3, S4, S40, S5,S6, S7, S10, S12, S13 and S16 were cloned and characterized. [The nucleotide sequences reported in this paper have been submitted to the EMBL/GenBank database under the following accession numbers: PaSFB1(AY805048), PaSFB2 (AY805049), PaSFB3 (AY805057),PaSFB4 (AY649872), PaSFB40 (AY649873), PaSFB5(AY805050), PaSFB6 (AY805051), PaSFB7 (AY805052),PaSFB10 (AY805053), PaSFB12 (AY805054), PaSFB13(AY805055), PaSFB16 (AY805056).] Though the coding regions of six of these alleles have been reported previously,the intron sequence has previously been reported only for S6. Analysis of the introns revealed sequence and length polymorphisms. A novel, PCR-based method to genotype cultivars and wild accessions was developed which combines fluorescently labelled primers amplifying the intron of SFB with similar primers for the first intron of S-RNase alleles. Intron length polymorphisms were then ascertained using a semi-automated sequencer. The convenience and reliability of this method for the determination of the self-incompatibility (SI) genotype was demonstrated both in sweet cherry cultivars representing alleles S1 to S16 and in individuals from a wild population encompassing S-alleles S17 to S22. This method will greatly expedite SI characterisation in sweet cherry and also facilitate large-scale studies of self-incompatibility in wild cherry and other Prunus populations.

  19. Characteristics of Phomopsis sp. isolates of plum trees origin

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    Živković Svetlana T.

    2007-01-01

    Full Text Available Twelve isolates of Phomopsis sp. were obtained from the branches and the trunk of plums (Prunus domestica L with decay symptoms in Valjevo, Ljig Koceljeva and Ub vicinity during 2004-2006. Morphological, pathogenic and growing characteristics were studied. Pathogen caused tissue necrosis of branches around the inoculate seats, and wrinkling and watering of plum fruits. All media were suitable for pathogen development, except prune agar. The best growth of isolates was at medium pH 5,5. The optimal temperature for growth and germination of pycnidiospores was 25°C.

  20. Mutation scan screening of Echinococcus granulosus isolates of Indian origin.

    Science.gov (United States)

    Bhattacharya, D; Pan, D; Bera, A K; Konar, A; Das, S K

    2008-08-01

    During the present investigation a total of forty Indian animal isolates were screened by single strand conformation polymorphism (SSCP) collected from sheep, goat, cattle and buffalo. The result of the study indicated that nuclear variants of Echinococcus granulosus were present in both small and large ruminants. SSCP phenotypes of AgB, intron of actin II and Hbx-2 have been deduced. Presence of nuclear variants due to mutation of E. granulosus has been discussed depending on hypotheses imparted earlier in literature. High polymophism of AgB demands further investigation because the gene is related with immune evasion and infectivity. This communication reports for the first time the comparative profile of Indian goat, sheep, cattle and buffalo isolates of E. granulosus complex.

  1. Lama glama con signología y lesiones compatibles con paratuberculosis causadas por Mycobacterium avium subespecie avium Lama glama with signology and lesion compatible with paratuberculosis and injuries caused by Mycobacterium avium subspecies avium

    Directory of Open Access Journals (Sweden)

    M.C Jorge

    2008-06-01

    Full Text Available Los camélidos sudamericanos (CS incluyen cuatro especies, guanaco, vicuña, alpaca y llama (Lama glama. En Argentina las llamas eran consideradas fauna y actualmente ganado, revalorizando su carne, fibra, cueros y pieles, también son un medio de subsistencia. Los CS son susceptibles a las enfermedades ocasionadas por micobacterias. El diagnóstico presuntivo se realiza por los signos clínicos y los hallazgos de necropsia y se confirma por técnicas bacteriológicas, moleculares e histopatología. El objetivo de este trabajo es describir un caso clínico con signos compatibles de paratuberculosis y el diagnóstico de laboratorio en una llama en cautiverio perteneciente a un zoológico de Olavarría, Provincia de Buenos Aires. En la necropsia se observaron lesiones granulomatosas en yeyuno, íleon y linfonodos mesentéricos compatibles con paratuberculosis, en los frotis directos y en la histopatología se observaron bacilos ácido-alcohol resistentes en cluster. Se confirmó la presencia de Mycobacterium avium subespecie avium por bacteriología y por PCR fue detectada la IS1245 característica de este agente, no detectando la IS900 correspondiente a Mycobacterium avium subespecie paratuberculosis. Esto permitió arribar al diagnóstico etiológico, combinando técnicas, de un caso de enteritis granulomatosa en llamas causado por Mycobacterium avium subespecie avium con signología y lesiones compatibles con paratuberculosis.Guanaco, vicuña, alpaca and llama (Lama glama are also known as Sudamerican camelids (SC. In Argentina llama was considered non profitable wildlife specie but now it is considered a mean for surviving because their meat, wool, leather and skin is valuable. SC are susceptible hosts of mycobacterial infections. A presumptive diagnosis is based on clinical and necropsy findings and is confirmed with bacterial isolation, molecular identification and histopathology. The objective of this publication is to describe a clinical

  2. Application of the C18-Carboxypropylbetaine Specimen Processing Method to Recovery of Mycobacterium avium subsp. paratuberculosis from Ruminant Tissue Specimens

    OpenAIRE

    Thornton, Charles G.; MacLellan, Kerry M.; Judith R Stabel; Carothers, Christine; Whitlock, Robert H.; Passen, Selvin

    2002-01-01

    The causative agent of Johne's disease is Mycobacterium avium subsp. paratuberculosis. This is a chronic, debilitating gastrointestinal disorder that affects ruminants and is responsible for significant economic loss. The specimen processing method that combines C18-carboxypropylbetaine (CB-18) treatment and lytic enzyme decontamination has been shown to improve the diagnosis of mycobacterioses. This processing method was applied to the isolation of M. avium subsp. paratuberculosis from rumin...

  3. Typing of clinical Mycobacterium avium complex strains cultured during a 2-year period in Denmark by using IS1245

    DEFF Research Database (Denmark)

    Bauer, Jeanett; Andersen, Åse B.; Askgaard, Dorthe;

    1999-01-01

    In the present study restriction fragment length polymorphism analyses with the recently described insertion sequence IS1245 as a probe was performed with clinical Mycobacterium avium complex strains cultured in Denmark during a 2-year period. The overall aim of the study was to disclose potential...... as potting soil) and veterinary samples were found to contain viable M avium isolates belonging to genotypes also found in humans....

  4. Virulence genotypes of Escherichia coli canine isolates from pyometra, cystitis and fecal origin.

    Science.gov (United States)

    Mateus, Luisa; Henriques, Sofia; Merino, Carolina; Pomba, Constança; Lopes da Costa, Luís; Silva, Elisabete

    2013-10-25

    Pyometra is the most common diestrual uterine disease of bitches. Escherichia coli is the most frequent bacterium isolated from the uterine content of pyometra uteri and it is associated with the most severe clinical signs, leading to endotoxemia and sepsis. In this study, canine E. coli isolates from pyometra (n=31), cystitis (n=23) and fecal (n=26) origin were compared regarding the prevalence of 23 potential virulence traits (15 virulence factor (VF) genes and 8 pathogenicity associated islands-PAIs), detected by PCR assays. Overall, there was a considerable overlap between pyometra, cystitis and fecal isolates regarding the phylogenetic grouping and virulence traits. Virulence traits more prevalent in pyometra than in cystitis and fecal isolates included two PAIs (PAI IV536 and PAI ICFT073) and three VF genes (sfa/focDE, fyuA and chuA). Regardless the isolates' origin, the average number of virulence traits per strain was higher in B2 than in the other phylogenetic groups (A, B1 and D). The prevalence of phylogenetic group B2 was significantly higher in pyometra (94%) than in cystitis (48%) and fecal (39%) isolates. In conclusion, pyometra isolates have a high potential of virulence and a broad virulence genotype, although being similar to a subset of cystitis and fecal isolates. This leads to the suggestion that cystitis and fecal isolates may be able to induce pyometra in receptive hosts.

  5. Prevalence and Antimicrobial Resistance of Salmonella Isolated from Animal-Origin Food Items in Gondar, Ethiopia

    Science.gov (United States)

    Garedew, Legesse; Alebachew, Zabishwork; Worku, Walelgn

    2016-01-01

    Salmonella has been found to be the major cause of foodborne diseases and a serious public health problem in the world, with an increasing concern for the emergence and spread of antimicrobial-resistant strains. A cross-sectional study was conducted between February 2014 and December 2015 on food items of animal origin to assess the prevalence and antimicrobial resistance profiles of Salmonella isolates using standard bacteriological methods. The overall prevalence rate of 5.5% was recorded from the total analyzed food items of animal origin. Salmonella isolates were detected from 12% of raw meat, 8% of minced meat, 2.9% of burger samples, 18% of raw eggs, and 6% of raw milk. Furthermore, antimicrobial susceptibility test identified 47.6% resistant Salmonella isolates, 28.6% intermediately sensitive isolates, and 23.8% susceptible isolates. Among Salmonella isolates tested, 42.6%, 28.6%, and 14.3% were found to be relatively resistant to tetracycline, sulfamethoxazole-trimethoprim, and ampicillin, respectively, while 9.5%–19% were intermediately resistant to tetracycline, amoxicillin, ampicillin, cephalothin, and nitrofurantoin. Therefore, our findings provide the prevalence and drug resistance of Salmonella from foods of animal origin and contribute information to scientists as well as public health researchers to minimize the prevalent and resistant foodborne Salmonella species in Ethiopia. PMID:28074185

  6. Prevalence and Antimicrobial Resistance of Salmonella Isolated from Animal-Origin Food Items in Gondar, Ethiopia.

    Science.gov (United States)

    Ejo, Mebrat; Garedew, Legesse; Alebachew, Zabishwork; Worku, Walelgn

    2016-01-01

    Salmonella has been found to be the major cause of foodborne diseases and a serious public health problem in the world, with an increasing concern for the emergence and spread of antimicrobial-resistant strains. A cross-sectional study was conducted between February 2014 and December 2015 on food items of animal origin to assess the prevalence and antimicrobial resistance profiles of Salmonella isolates using standard bacteriological methods. The overall prevalence rate of 5.5% was recorded from the total analyzed food items of animal origin. Salmonella isolates were detected from 12% of raw meat, 8% of minced meat, 2.9% of burger samples, 18% of raw eggs, and 6% of raw milk. Furthermore, antimicrobial susceptibility test identified 47.6% resistant Salmonella isolates, 28.6% intermediately sensitive isolates, and 23.8% susceptible isolates. Among Salmonella isolates tested, 42.6%, 28.6%, and 14.3% were found to be relatively resistant to tetracycline, sulfamethoxazole-trimethoprim, and ampicillin, respectively, while 9.5%-19% were intermediately resistant to tetracycline, amoxicillin, ampicillin, cephalothin, and nitrofurantoin. Therefore, our findings provide the prevalence and drug resistance of Salmonella from foods of animal origin and contribute information to scientists as well as public health researchers to minimize the prevalent and resistant foodborne Salmonella species in Ethiopia.

  7. Isolated Anomalous Origin of Left Pulmonary Artery From the Descending Aorta: An Embryologic Ambiguity.

    Science.gov (United States)

    Gnanappa, Ganesh Kumar; Laohachai, Karina; Orr, Yishay; Ayer, Julian

    2016-11-01

    Anomalous origin of a branch pulmonary artery from the aorta is a rare malformation, accounting for 0.12% of all congenital heart defects. Anomalous origin of the left pulmonary artery from the aorta (ALPA) constitutes a small proportion of these cases. ALPA has been reported to arise from the ascending aorta with various embryologic postulates. We report a case of isolated ALPA arising from the descending aorta in association with a patent ductus arteriosus, to emphasize its embryologic ambiguity.

  8. Multilocus sequence typing indicates diverse origins of invasive Candida tropicalis isolates in China

    Institute of Scientific and Technical Information of China (English)

    Fan Xin; Xiao Meng; Wang He; Zhang Li; Kong Fanrong; Lu Juan; Hu Zhidong

    2014-01-01

    Background According to data from the China Hospital Invasive Fungal Surveillance Net (CHIF-NET) 2010,Candida tropicalis (C.tropicalis) is the third most common pathogen causing invasive candidiasis.Moreover,the majority of fluconazole-resistant C.tropicalis isolates were from a single hospital.Therefore,a molecular epidemiological survey is necessary to investigate the genetic relatedness of C.tropicalis isolates in China.Methods In this study,48 C.tropicalis isolates causing invasive fungal infections from four tertiary hospitals in China were studied.All the isolates were identified by sequencing the internal transcribed spacer region.Antifungal susceptibility to triazoles,amphotericin B,and caspofungin was determined by the Clinical and Laboratory Standards Institute standard broth microdilution method.Multilocus sequence typing (MLST) was performed,and phylogenetic analysis was further performed by the eBURST and maximum parsimony (MP) methods to characterize the genetic relatedness of isolates.Results MLST discriminated 40 diploid sequence types (DSTs) among 48 isolates,including 36 novel DSTs,and the XYR1 gene showed the highest discriminatory power.The DSTs obtained from this study were compared with those of previously reported C.tropicalis isolates,and there was poor type alignment with regional strains.Nine groups and 11 singletons were identified by eBURST,whereas two groups and 10 subgroups were clustered by MP analysis.Generally,there were no obvious correlations between clonal clusters generated and the specimen source or hospital origin.Seven fluconazole-resistant isolates were confirmed and assigned to three distinguishable branches.Conclusions The results suggested diverse origins of invasive C.tropicalis isolates in China.Although most invasive C.tropicalis strains in the mainland of China were clustered with previously characterized Asian isolates,major C.tropicalis clusters identified in this study were genetically distinct from those of other

  9. Antimicrobial-producing wild lactococci isolated from artisanal and non-dairy origins

    NARCIS (Netherlands)

    Ayad, E.H.E.; Verheul, A.; Wouters, J.T.M.; Smit, G.

    2002-01-01

    Wild lactococcal strains with diverse properties isolated from dairy and non-dairy origins and characterised by their flavour-forming abilities were tested for antagonistic activities to assess their potential for use as new starter cultures. Thirty-two of seventy-nine strains examined inhibited the

  10. Genome Sequence of the "Indian Bison Type" Biotype of Mycobacterium avium subsp. paratuberculosis Strain S5.

    Science.gov (United States)

    Singh, Shoor Vir; Kumar, Naveen; Singh, Shree Narayan; Bhattacharya, Tapas; Sohal, Jagdip Singh; Singh, Pravin Kumar; Singh, Ajay Vir; Singh, Brajesh; Chaubey, Kundan Kumar; Gupta, Saurabh; Sharma, Nitu; Kumar, Shailesh; Raghava, Gajendra Pal Singh

    2013-01-01

    We report the 4.79-Mb genome sequence of the "Indian Bison Type" biotype of Mycobacterium avium subsp. paratuberculosis strain S5, isolated from a terminally sick Jamunapari goat at the CIRG (Central Institute for Research on Goats) farm in India. This draft genome will help in studying novelties of this biotype, which is widely distributed in animals and human beings in India.

  11. Cloning and molecular characterisation of resuscitation promoting factor-like gene from Mycobacterium avium subspecies avium

    Directory of Open Access Journals (Sweden)

    R Kavitha

    2016-01-01

    Full Text Available Purpose: Resuscitation promoting factor (Rpf-like gene of Mycobacterium avium subspecies paratuberculosis has been known to stimulate the growth of mycobacteria and enhances the recovery of replicating cells from non-replicating phases. The objective of the study was to produce recombinant rpf-like protein of M. avium subspecies avium protein for purification and physico-chemical characterisation. Materials and Methods: The identified rpf gene of M. avium subspecies avium was cloned, subcloned, sequenced and expressed in Escherichia coli expression system for the production of the recombinant protein. The expressed recombinant Rpf protein was confirmed by Western blot and the extract was purified to yield a pure recombinant protein. Results: An rpf-like gene of 675 bp size in the M. avium subspecies avium was identified. This gene was expressed and the recombinant Rpf weighed 65 kDa as confirmed by Western blot. The M. avium recombinant Rpf protein was extracted under denatured conditions and purified yielding a recombinant protein with >90% purity. Conclusions: Identification, cloning, sequencing and expression of a rpf-like gene from M. avium suggest that RpfA is present in this species also, which might be involved in reactivation phenomenon in this high-risk pathogen.

  12. Characterization of temperate phages infecting Clostridium difficile isolates of human and animal origins.

    Science.gov (United States)

    Sekulovic, Ognjen; Garneau, Julian R; Néron, Audrey; Fortier, Louis-Charles

    2014-04-01

    Clostridium difficile is a Gram-positive pathogen infecting humans and animals. Recent studies suggest that animals could represent potential reservoirs of C. difficile that could then transfer to humans. Temperate phages contribute to the evolution of most bacteria, for example, by promoting the transduction of virulence, fitness, and antibiotic resistance genes. In C. difficile, little is known about their role, mainly because suitable propagating hosts and conditions are lacking. Here we report the isolation, propagation, and preliminary characterization of nine temperate phages from animal and human C. difficile isolates. Prophages were induced by UV light from 58 C. difficile isolates of animal and human origins. Using soft agar overlays with 27 different C. difficile test strains, we isolated and further propagated nine temperate phages: two from horse isolates (ΦCD481-1 and ΦCD481-2), three from dog isolates (ΦCD505, ΦCD506, and ΦCD508), and four from human isolates (ΦCD24-2, ΦCD111, ΦCD146, and ΦCD526). Two phages are members of the Siphoviridae family (ΦCD111 and ΦCD146), while the others are Myoviridae phages. Pulsed-field gel electrophoresis and restriction enzyme analyses showed that all of the phages had unique double-stranded DNA genomes of 30 to 60 kb. Phages induced from human C. difficile isolates, especially the members of the Siphoviridae family, had a broader host range than phages from animal C. difficile isolates. Nevertheless, most of the phages could infect both human and animal strains. Phage transduction of antibiotic resistance was recently reported in C. difficile. Our findings therefore call for further investigation of the potential risk of transduction between animal and human C. difficile isolates.

  13. Isolation of restriction fragments containing origins of replication from complex genomes.

    Science.gov (United States)

    Mesner, Larry D; Hamlin, Joyce L

    2015-01-01

    The identification and isolation of origins of replication from mammalian genomes has been a demanding task owing to the great complexity of these genomes. However, two methods have been refined in recent years each of which allows significant enrichment of recently activated origins of replication from asynchronous cell cultures. In one of these, nascent strands are melted from the long template DNA, and the small, origin-centered strands are isolated on sucrose gradients. The second method involves the selective entrapment of bubble-containing fragments in gelling agarose and their subsequent recovery and isolation by molecular cloning. Libraries prepared by this method from Chinese hamster and human cells have been shown to be extremely pure, and provide a renewable resource of origins that can be used as probes on microarrays or sequenced by high-throughput techniques to localize them within the genomic source. The bubble-trapping method is described here for asynchronous mammalian cells that grow with reasonable doubling times and from which nuclear matrices can be reliably prepared. The method for nuclear matrix preparation and enrichment of replication intermediates is described in an accompanying chapter entitled "Purification of restriction fragments containing replication intermediates from mammalian cells for 2-D gel analysis" (Chapter 16 ).

  14. ERIC-PCR Genotyping of Some Campylobacter jejuni Isolates of Chicken and Human Origin in Egypt.

    Science.gov (United States)

    Ahmed, Heba A; El Hofy, Fatma I; Ammar, Ahmed M; Abd El Tawab, Ashraf A; Hefny, Ahmed A

    2015-12-01

    The public health importance of the genus Campylobacter is attributed to several species causing diarrhea in consumers. Poultry and their meat are considered the most important sources of human campylobacteriosis. In this study, 287 samples from chicken (131 cloacal swabs, 39 chicken skin, 78 chicken meat, and 39 cecal parts) obtained from retail outlets as well as 246 stool swabs from gastroenteritis patients were examined. A representative number of the biochemically identified Campylobacter jejuni isolates were identified by real-time PCR, confirming the identification of the isolates as C. jejuni. Genotyping of the examined isolates (n = 31) by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) revealed a high discriminatory index of ERIC-PCR (D = 0.948), dividing C. jejuni isolates of chicken and human origins into 18 profiles and four clusters. The 18 profiles obtained indicated the heterogeneity of C. jejuni. Dendrogram analysis showed that four clusters were generated; all human isolates fell into clusters I and III. These observations further support the existence of a genetic relationship between human and poultry isolates examined in the present study. In conclusion, the results obtained support the speculation that poultry and poultry meat have an important role as sources of infection in the acquisition of Campylobacter infection in humans.

  15. Characterization of Haemolysin of Staphylococcus aureus Isolated from Food of Animal Origin

    Directory of Open Access Journals (Sweden)

    Dwi Ariyanti

    2015-11-01

    Full Text Available Staphylococcus aureus is an important pathogen bacteria causing food poisoning and various infection in animals and humans. Haemolysin is one of the virulence factors of Staphylococcus aureus. The aims of the research were to characterize haemolysins of Staphylococcus aureus isolated from various food of animal origin, phenotypic- and genotypically. In the present study, eleven Staphylococcus aureus isolated from various food of animal origins from traditional markets and supermarkets in Yogyakarta, Sidoarjo, Jakarta, and Bandung were characterized for haemolysin, pheno- and genotypically. Characterization of haemolysin phenotypically based on haemolysis pattern of Staphylococcus aureus on sheep blood agar plate. Genes encoding hemolysin were amplified with specific primers by using polymerase chain reaction (PCR technique. The results of the studies showed that Staphylococcus aureus on sheep blood agar plates revealed an alpha haemolysis pattern (18,18%, beta haemolysis (27,27% and gamma haemolysis (54,55%. Based on amplification of the gene encoding haemolysin of Staphylococcus aureus with specific primers showed hla genes (81,81%, and hla combined with hlb genes (18,18%. The amplification of gene hla and hlb had a single amplicon with a size of approximately 534 bp and 833 bp, respectively. The haemolysin characteristics of Staphylococcus aureus from various food of animal origin could be used as important information to control staphylococcal food poisoning.Keywords : Staphylococcus aureus, haemolysin, PCR, food of animal origins

  16. Infectious hematopoietic necrosis virus: monophyletic origin of European isolates from North American genogroup M.

    Science.gov (United States)

    Enzmann, P J; Kurath, G; Fichtner, D; Bergmann, S M

    2005-09-23

    Infectious hematopoietic necrosis virus (IHNV) was first detected in Europe in 1987 in France and Italy, and later, in 1992, in Germany. The source of the virus and the route of introduction are unknown. The present study investigates the molecular epidemiology of IHNV outbreaks in Germany since its first introduction. The complete nucleotide sequences of the glycoprotein (G) and non-virion (NV) genes from 9 IHNV isolates from Germany have been determined, and this has allowed the identification of characteristic differences between these isolates. Phylogenetic analysis of partial G gene sequences (mid-G, 303 nucleotides) from North American IHNV isolates (Kurath et al. 2003) has revealed 3 major genogroups, designated U, M and L. Using this gene region with 2 different North American IHNV data sets, it was possible to group the European IHNV strains within the M genogroup, but not in any previously defined subgroup. Analysis of the full length G gene sequences indicated that an independent evolution of IHN viruses had occurred in Europe. IHN viruses in Europe seem to be of a monophyletic origin, again most closely related to North American isolates in the M genogroup. Analysis of the NV gene sequences also showed the European isolates to be monophyletic, but resolution of the 3 genogroups was poor with this gene region. As a result of comparative sequence analyses, several different genotypes have been identified circulating in Europe.

  17. Hepatitis B Virus Genotype D Isolates Circulating in Chapecó, Southern Brazil, Originate from Italy

    Science.gov (United States)

    Gusatti, Carolina Souza; Costi, Cintia; Halon, Maria Laura; Grandi, Tarciana; Medeiros, Arlete Ferrari Rech; Silva, Cláudia Maria Dornelles; Gomes, Selma Andrade; Silva, Marcia Susana Nunes; Niel, Christian; Rossetti, Maria Lucia Rosa

    2015-01-01

    Hepatitis B virus genotype A1 (HBV/A1), of African origin, is the most prevalent genotype in Brazil, while HBV/F predominates in the other South American countries. However, HBV/D is the most common in the three states of southern Brazil, where ‘islands’ of elevated prevalence, as Chapecó and other cities, have been described. In this study, 202 HBV chronic carriers attending in 2013 the viral hepatitis ambulatory of Chapecó, were investigated. In comparison with previous studies performed in the same ambulatory, a rapid aging of the HBV infected population was observed (mean age of the newly diagnosed patients increasing from 29.9 ± 10.3 years in 1996 to 44.4 ± 13.3 years in 2013), probably due to a singular vaccination schedule at Chapecó that included not only children but also adolescents. Phylogenetic and BLAST analyses (S region) classified 91 HBV isolates into genotypes A (n = 3) and D (n = 88). The majority of HBV/D isolates were closely related to D3 sequences. To understand the reasons for the absence or near absence of genotypes A and F, and how HBV/D was introduced in the south of Brazil, HBV/D infected patients were inquired about their genealogical and geographical origins. Forty-three (52%) patients have their four grandparents of Italian origin, vs. seven (8%) who have their four grandparents of Brazilian origin. At all, 65 out of 83 (78%) patients had at least one grandparent originating from Italy. Taking into consideration the fact that Italy is one of the few countries where subgenotype D3 is predominant, the results strongly suggested that HBV/D was introduced in Brazil through Italian immigration which culminated between 1870 and 1920. PMID:26275046

  18. Hepatitis B Virus Genotype D Isolates Circulating in Chapeco, Southern Brazil, Originate from Italy.

    Directory of Open Access Journals (Sweden)

    Carolina Souza Gusatti

    Full Text Available Hepatitis B virus genotype A1 (HBV/A1, of African origin, is the most prevalent genotype in Brazil, while HBV/F predominates in the other South American countries. However, HBV/D is the most common in the three states of southern Brazil, where 'islands' of elevated prevalence, as Chapecó and other cities, have been described. In this study, 202 HBV chronic carriers attending in 2013 the viral hepatitis ambulatory of Chapecó, were investigated. In comparison with previous studies performed in the same ambulatory, a rapid aging of the HBV infected population was observed (mean age of the newly diagnosed patients increasing from 29.9 ± 10.3 years in 1996 to 44.4 ± 13.3 years in 2013, probably due to a singular vaccination schedule at Chapecó that included not only children but also adolescents. Phylogenetic and BLAST analyses (S region classified 91 HBV isolates into genotypes A (n = 3 and D (n = 88. The majority of HBV/D isolates were closely related to D3 sequences. To understand the reasons for the absence or near absence of genotypes A and F, and how HBV/D was introduced in the south of Brazil, HBV/D infected patients were inquired about their genealogical and geographical origins. Forty-three (52% patients have their four grandparents of Italian origin, vs. seven (8% who have their four grandparents of Brazilian origin. At all, 65 out of 83 (78% patients had at least one grandparent originating from Italy. Taking into consideration the fact that Italy is one of the few countries where subgenotype D3 is predominant, the results strongly suggested that HBV/D was introduced in Brazil through Italian immigration which culminated between 1870 and 1920.

  19. Hepatitis B Virus Genotype D Isolates Circulating in Chapecó, Southern Brazil, Originate from Italy.

    Science.gov (United States)

    Gusatti, Carolina Souza; Costi, Cintia; Halon, Maria Laura; Grandi, Tarciana; Medeiros, Arlete Ferrari Rech; Silva, Cláudia Maria Dornelles; Gomes, Selma Andrade; Silva, Marcia Susana Nunes; Niel, Christian; Rossetti, Maria Lucia Rosa

    2015-01-01

    Hepatitis B virus genotype A1 (HBV/A1), of African origin, is the most prevalent genotype in Brazil, while HBV/F predominates in the other South American countries. However, HBV/D is the most common in the three states of southern Brazil, where 'islands' of elevated prevalence, as Chapecó and other cities, have been described. In this study, 202 HBV chronic carriers attending in 2013 the viral hepatitis ambulatory of Chapecó, were investigated. In comparison with previous studies performed in the same ambulatory, a rapid aging of the HBV infected population was observed (mean age of the newly diagnosed patients increasing from 29.9 ± 10.3 years in 1996 to 44.4 ± 13.3 years in 2013), probably due to a singular vaccination schedule at Chapecó that included not only children but also adolescents. Phylogenetic and BLAST analyses (S region) classified 91 HBV isolates into genotypes A (n = 3) and D (n = 88). The majority of HBV/D isolates were closely related to D3 sequences. To understand the reasons for the absence or near absence of genotypes A and F, and how HBV/D was introduced in the south of Brazil, HBV/D infected patients were inquired about their genealogical and geographical origins. Forty-three (52%) patients have their four grandparents of Italian origin, vs. seven (8%) who have their four grandparents of Brazilian origin. At all, 65 out of 83 (78%) patients had at least one grandparent originating from Italy. Taking into consideration the fact that Italy is one of the few countries where subgenotype D3 is predominant, the results strongly suggested that HBV/D was introduced in Brazil through Italian immigration which culminated between 1870 and 1920.

  20. Antibiotic resistances in Listeria monocytogenes and Salmonella enterica isolated from foods with animal origin

    Directory of Open Access Journals (Sweden)

    Baltasar Balsalobre Hernández

    2004-12-01

    Full Text Available Extensive use of antibiotics in both human and animal health and in cattle production has generated resistant microorganisms to common antibiotics. Resistances spread caused by human and animal therapeutic is well known, but we know poorly frecuency of resistant bacteria in foods with animal origin and destinated to human consumers. In this paper, sensitivity to nineteen antibiotics was investigated in Listeria monocytogenes and Salmonella enterica strains isolated from foods with animal origin, including fresh meat, hamburgers, fresh sausages, boiled ham and new-laid chicken eggs. The plate diffusion method of Bauer-Kirby was used.Listeria monocytogenes strains showed a very high sensitivity to all antibiotics checked, with the exception of one strain tetracycline resistant. In contrast, Salmonella enterica showed a high frecuency of resistances, in special to tetracycline, streptomycin, nalidixic acid, ticarcillin, ampicillin and chloramphenicol. Moreover, multi-resistance was a common phenomenon. Twenty percent of S. enterica strains were resistant to four or more antibiotics. Frecuency of resistances was higher in 4,5,12:i:-, Hadar, Typhimurium and Virchow serotypes.In conclusion, Salmonella enterica strains isolated from foods with animal origin and destinated to human consumers are usually resistant to several antibiotics. The significance of this observation and its potential health risk must be investigated.

  1. CD4 T Cell Dependent Colitis Exacerbation Following Re-Exposure of Mycobacterium avium ssp. paratuberculosis

    Science.gov (United States)

    Suwandi, Abdulhadi; Bargen, Imke; Pils, Marina C.; Krey, Martina; Zur Lage, Susanne; Singh, Anurag K.; Basler, Tina; Falk, Christine S.; Seidler, Ursula; Hornef, Mathias W.; Goethe, Ralph; Weiss, Siegfried

    2017-01-01

    Mycobacterium avium ssp. paratuberculosis (MAP) is the causative agent of Johne's disease (JD), a chronic inflammatory bowel disease of cattle characterized by intermittent to chronic diarrhea. In addition, MAP has been isolated from Crohn's disease (CD) patients. The impact of MAP on severity of clinical symptoms in JD as well as its role in CD are yet unknown. We have previously shown that MAP is able to colonize inflamed enteric tissue and to exacerbate the inflammatory tissue response (Suwandi et al., 2014). In the present study, we analyzed how repeated MAP administration influences the course of dextran sulfate sodium (DSS)-induced colitis. In comparison to mice exposed to DSS or MAP only, repeated exposure of DSS-treated mice to MAP (DSS/MAP) revealed a significantly enhanced clinical score, reduction of colon length as well as severe CD4+ T cell infiltration into the colonic lamina propria. Functional analysis identified a critical role of CD4+ T cells in the MAP-induced disease exacerbation. Additionally, altered immune responses were observed when closely related mycobacteria species such as M. avium ssp. avium and M. avium ssp. hominissuis were administered. These data reveal the specific ability of MAP to aggravate intestinal inflammation and clinical symptoms. Overall, this phenotype is compatible with similar disease promoting capabilites of MAP in JD and CD.

  2. Optimization of methods for the detection of Mycobacterium avium subsp. paratuberculosis in milk and colostrum of naturally infected dairy cows

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is primarily shed into the feces but it has also been isolated from the milk and colostrum of cows. Because of this, there exists concern about transfer of the organism from dam to calf and about the prevalence of MAP in the milk supply. The prevalen...

  3. Mycobacterium avium Complex Infection in a Patient with Sickle Cell Disease and Severe Iron Overload

    Directory of Open Access Journals (Sweden)

    Kamal Shemisa

    2014-01-01

    Full Text Available A 34-year-old female with sickle cell anemia (hemoglobin SS disease and severe iron overload presented to our institution with the subacute presentation of recurrent pain crisis, fever of unknown origin, pancytopenia, and weight loss. A CT scan demonstrated both lung and liver nodules concerning for granulomatous disease. Subsequent biopsies of the liver and bone marrow confirmed the presence of noncaseating granulomas and blood cultures isolated Mycobacterium avium complex MAC. Disseminated MAC is considered an opportunistic infection typically diagnosed in the immunocompromised and rarely in immunocompetent patients. An appreciable number of mycobacterial infection cases have been reported in sickle cell disease patients without immune dysfunction. It has been reported that iron overload is known to increase the risk for mycobacterial infection in vitro and in vivo studies. While iron overload is primarily known to cause end organ dysfunction, the clinical relationship with sickle cell disease and disseminated MAC infection has not been reported. Clinical iron overload is a common condition diagnosed in the sub-Saharan African population. High dietary iron, genetic defects in iron trafficking, as well as hemoglobinopathy are believed to be the etiologies for iron overload in this region. Patients with iron overload in this region were 17-fold more likely to die from Mycobacterium tuberculosis. Both experimental and clinical evidence suggest a possible link to iron overload and mycobacterial infections; however larger observational studies are necessary to determine true causality.

  4. Mycobacterium avium subsp. Paratuberculosis (MAP) as a modifying factor in Crohn's disease.

    LENUS (Irish Health Repository)

    Sibartie, Shomik

    2010-02-01

    Crohn\\'s disease (CD) is a multifactorial syndrome with genetic and environmental contributions. Mycobacterium avium subspecies paratuberculosis (MAP) has been frequently isolated from mucosal tissues of patients with CD but the cellular immune response to this bacterium has been poorly described. Our aim was to examine the influence of MAP on T-cell proliferation and cytokine responses in patients with inflammatory bowel disease (IBD).

  5. Antimicrobial susceptibility profiles of Staphylococcus aureus isolates classified according to their origin in a tertiary hospital in Korea.

    Science.gov (United States)

    Moon, Hee-Won; Kim, Hyun Jung; Hur, Mina; Yun, Yeo-Min

    2014-12-01

    We performed a comprehensive analysis on 3,594 Staphylococcus aureus isolates from routine culture during the last 4 years. The antimicrobial susceptibilities of the isolates were analyzed according to their origin and were compared based on the type of specimens. The proportion of methicillin-resistant Staphylococcus aureus (MRSA) in community-associated (CA), health care-associated, community onset (HACO), and health care-associated (HA) isolates were 33.0%, 54.3%, and 73.3%, respectively. The MRSA rate differed significantly between specimens, with the highest rate from urine in the CA and HACO isolates, whereas the highest rate from the respiratory tract was in the HA isolates. The monitoring of the MRSA rate in CA, HACO, and HA S aureus isolates would be valuable for surveillance. The elevated rates of MRSA in urinary specimens from CA and HCA isolates need to be addressed for infection control.

  6. Biological entities isolated from the stratosphere (22-27km): case for their space origin

    Science.gov (United States)

    Wainwright, Milton; Rose, Christopher E.; Baker, Alexander J.; Wickramasinghe, N. Chandra

    2013-09-01

    Biological entities were isolated at a height of between 22-27 km in the stratosphere. Sampling of this region was carried out in the UK in July 2013 using a relatively simple low-cost balloon-borne sampler carrying aseptically clean scanning electron microscope stubs onto which aerosols were directly captured. The entities varied from a presumptive colony of ultra-small bacteria to two unusual individual organisms - part of a diatom frustule and a 200 micron-sized particle mass interlaced with biological filaments. Biological entities of this nature have not previously been reported occurring in the stratosphere; their likely origin is discussed and we provide arguments to support our view that such biological entities may have arrived from space. The new data gives strong confirmation of the Hoyle-Wickramasinghe theory of cometary panspermia.

  7. Characterization and possible origins of isolated douglas fir stands on the Colorado Plateau

    Energy Technology Data Exchange (ETDEWEB)

    Spence, J.R. [National Park Service, Page, AZ (United States)

    1995-09-01

    The floristic composition of several isolated stands of Pseudotsuga menziesii on the Colorado Plateau is compared. All occurred between 1700-2000 meters, which was about 300-500 meters below typical lower limits for the species. Most stands showed evidence of reproduction and recruitment. A small widespread group of species existed that was common to all stands, but beta diversity was high. A preliminary analysis of possible origins of the stands suggested that they are relictual from the late Wisconsin or early Holocene. Bird-dispersed species were less prevalent than expected compared with existing high elevation conifer forests. These stands represent important resources for Quaternary research, but are extremely vulnerable to human-caused disturbances. It is recommended that they be provided protection from disturbance through appropriate management activities of the various land management agencies.

  8. Pre-Holocene Origin for the Coronopus navasii Disjunction: Conservation Implications from Its Long Isolation.

    Directory of Open Access Journals (Sweden)

    Sara Martín-Hernanz

    Full Text Available Integration of unexpected discoveries about charismatic species can disrupt their well-established recovery plans, particularly when this requires coordinate actions among the different governments responsible. The Critically Endangered Coronopus navasii (Brassicaceae was considered a restricted endemism to a few Mediterranean temporary ponds in a high mountain range of Southeast Spain, until a new group of populations were discovered 500 km North in 2006. Ten years after this finding, its management has not been accommodated due to limited information of the new populations and administrative inertia. In this study, DNA sequences and species distribution models are used to analyse the origin of the C. navasii disjunction as a preliminary step to reassess its recovery plan. Molecular results placed the disjunction during Miocene-Pleistocene (6.30-0.49 Mya, plastid DNA; 1.45-0.03 Mya, ribosomal DNA, which discards a putative human-mediated origin. In fact, the haplotype network and the low gene flow estimated between disjunct areas suggest long-term isolation. Dispersal is the most likely explanation for the disjunction as interpreted from the highly fragmented distribution projected to the past. Particularly, a northward dispersal from Southeast is proposed since C. navasii haplotype network is connected to the sister-group through the southern haplotype. Although the reassessment of C. navasii conservation status is more optimistic under the new extent of occurrence, its long-term survival may be compromised due to the: (1 natural fragmentation and rarity of the species habitat, (2 genetic isolation between the two disjunct areas, and (3 northward shift of suitable areas under future climate change scenarios. Several ex-situ and in-situ conservation measures are proposed for integrating Central East Spanish populations into the on-going recovery plan, which still only contemplates Southeast populations and therefore does not preserve the

  9. Mycobacterium avium restriction fragment lenght polymorphism-IS IS1245 and the simple double repetitive element polymerase chain reaction typing method to screen genetic diversity in Brazilian strains

    Directory of Open Access Journals (Sweden)

    Patrícia Carvalho de Sequeira

    2005-11-01

    Full Text Available Simple double repetitive element polymerase chain reaction (MaDRE-PCR and Pvu II-IS1245 restriction fragment length polymorphism (RFLP typing methods were used to type 41 Mycobacterium avium isolates obtained from 14 Aids inpatients and 10 environment and animals specimens identified among 53 mycobacteria isolated from 237 food, chicken, and pig. All environmental and animals strains showed orphan patterns by both methods. By MaDRE-PCR four patients, with multiple isolates, showed different patterns, suggesting polyclonal infection that was confirmed by RFLP in two of them. This first evaluation of MaDRE-PCR on Brazilian M. avium strains demonstrated that the method seems to be useful as simple and less expensive typing method for screening genetic diversity in M. avium strains on selected epidemiological studies, although with limitation on analysis identical patterns except for one band.

  10. Emphysematous pyometra secondary to Enterococcus avium infection in a dog.

    Science.gov (United States)

    Chang, An-Chi; Cheng, Ching-Chang; Wang, Hsien-Chi; Lee, Wei-Ming; Shyu, Ching-Lin; Lin, Cheng-Chung; Chen, Kuan-Sheng

    2016-06-16

    A 5-year-old female intact Mastiff dog was presented with a history of vaginal discharge for 1 day. Physical examination revealed a sanguineo-purulent vaginal discharge and systemic inflammatory response syndrome. Abdominal radiographs showed several dilated and gas- filled tubular loops. The differential diagnoses included emphysematous pyometra or small intestinal mechanical ileus. Surgical exploration of the abdomen demonstrated a severely dilated and gas-filled uterus, and emphysematous pyometra was confirmed. The patient's clinical signs resolved after ovariohysterectomy. Histopathology revealed mild endometrial cystic hyperplasia with infiltration of inflammatory cells in the superficial endometrial epithelia. Enterococcus avium, an α-hemolytic gram-positive coccus, was isolated from the uterus. This paper highlights the radiographic features of emphysematous pyometra and a pathogen that has never been reported to be associated with canine pyometra previously.

  11. Origins.

    Science.gov (United States)

    Online-Offline, 1999

    1999-01-01

    Provides an annotated list of resources dealing with the theme of origins of life, the universe, and traditions. Includes Web sites, videos, books, audio materials, and magazines with appropriate grade levels and/or subject disciplines indicated; professional resources; and learning activities. (LRW)

  12. Novel isolation strategy to deliver pure fetal-origin and maternal-origin mesenchymal stem cell (MSC) populations from human term placenta.

    Science.gov (United States)

    Patel, J; Shafiee, A; Wang, W; Fisk, N M; Khosrotehrani, K

    2014-11-01

    The placenta is an abundant source of mesenchymal stem/stromal cells (MSC). Although presumed of translationally-advantageous fetal origin, the literature instead suggests a high incidence of either contaminating or pure maternal MSC. Despite definitional criteria that MSC are CD34-, increasing evidence suggests that fetal MSC may be CD34 positive in vivo. We flow sorted term placental digests based on CD34+ expression and exploited differential culture media to isolate separately pure fetal and maternal MSC populations. This method has considerable translational implications, in particular to clinical trials underway with "placental" MSC of uncertain or decidual origin.

  13. Association between Mycobacterium avium subsp. paratuberculosis infection and culling in dairy cattle herds

    Directory of Open Access Journals (Sweden)

    R Arrazuría

    2014-01-01

    Full Text Available The present study was designed to analyse the causes for culling in dairy herds with different Mycobacterium avium subsp. paratuberculosis infection status and to compare these causes with those observed over the general dairy cattle population. During 2009, causes for culling were registered in two different groups of farms: (1 farms with seropositive cows for three consecutive years (2007-2009 but where Mycobacterium avium subsp. paratuberculosis has not been isolated from any of the fecal samples collected and (2 farms with Mycobacterium avium subsp. paratuberculosis seropositive cows for three consecutive years (2007-2009 and where the bacteria has been isolated from at least one fecal sample. Causes for animal loss were compared between both groups and between them and the general dairy cattle population by means of regression analysis. The distribution of culling reasons was different between infected herds (both bacteriologically positive and negative and the general population. The percentage of losses seemed to be higher in infected herds from the first parity on. The most remarkable difference among groups was observed in losses due to "death/urgent slaughter".

  14. Replication and long-term persistence of bovine and human strains of Mycobacterium avium subsp. paratuberculosis within Acanthamoeba polyphaga.

    Science.gov (United States)

    Mura, Manuela; Bull, Tim J; Evans, Hugh; Sidi-Boumedine, Karim; McMinn, Liz; Rhodes, Glenn; Pickup, Roger; Hermon-Taylor, John

    2006-01-01

    Free-living protists are ubiquitous in the environment and form a potential reservoir for the persistence of animal and human pathogens. Mycobacterium avium subsp. paratuberculosis is the cause of Johne's disease, a systemic infection accompanied by chronic inflammation of the intestine that affects many animals, including primates. Most humans with Crohn's disease are infected with this chronic enteric pathogen. Subclinical infection with M. avium subsp. paratuberculosis is widespread in domestic livestock. Infected animals excrete large numbers of robust organisms into the environment, but little is known about their ability to replicate and persist in protists. In the present study we fed laboratory cultures of Acanthamoeba polyphaga with bovine and human strains of M. avium subsp. paratuberculosis. Real-time PCR showed that the numbers of the pathogens fell over the first 4 to 8 days and recovered by 12 to 16 days. Encystment of the amoebic cultures after 4 weeks resulted in a 2-log reduction in the level of M. avium subsp. paratuberculosis, which returned to the original level by 24 weeks. Extracts of resection samples of human gut from 39 patients undergoing abdominal surgery were fed to cultures of A. polyphaga. M. avium subsp. paratuberculosis detected by nested IS900 PCR with amplicon sequencing and visualized by IS900 in situ hybridization and auramine-rhodamine staining was found in cultures derived from 13 of the patients and was still present in the cultures after almost 4 years of incubation. Control cultures were negative. M. avium subsp. paratuberculosis has the potential for long-term persistence in environmental protists.

  15. Genetic analysis of individual origins supports isolation of grizzly bears in the Greater Yellowstone Ecosystem

    Science.gov (United States)

    Haroldson, Mark A.; Schwartz, Charles; Kendall, Katherine C.; Gunther, Kerry A.; Moody, David S.; Frey, Kevin L.; Paetkau, David

    2010-01-01

    The Greater Yellowstone Ecosystem (GYE) supports the southernmost of the 2 largest remaining grizzly bear (Ursus arctos) populations in the contiguous United States. Since the mid-1980s, this population has increased in numbers and expanded in range. However, concerns for its long-term genetic health remain because of its presumed continued isolation. To test the power of genetic methods for detecting immigrants, we generated 16-locus microsatellite genotypes for 424 individual grizzly bears sampled in the GYE during 1983–2007. Genotyping success was high (90%) and varied by sample type, with poorest success (40%) for hair collected from mortalities found ≥1 day after death. Years of storage did not affect genotyping success. Observed heterozygosity was 0.60, with a mean of 5.2 alleles/marker. We used factorial correspondence analysis (Program GENETIX) and Bayesian clustering (Program STRUCTURE) to compare 424 GYE genotypes with 601 existing genotypes from grizzly bears sampled in the Northern Continental Divide Ecosystem (NCDE) (FST  =  0.096 between GYE and NCDE). These methods correctly classified all sampled individuals to their population of origin, providing no evidence of natural movement between the GYE and NCDE. Analysis of 500 simulated first-generation crosses suggested that over 95% of such bears would also be detectable using our 16-locus data set. Our approach provides a practical method for detecting immigration in the GYE grizzly population. We discuss estimates for the proportion of the GYE population sampled and prospects for natural immigration into the GYE.

  16. Improvement of the Original Isolation Procedure for Hormone Studies in Short-Time Culture

    Directory of Open Access Journals (Sweden)

    Mukadder Atmaca

    2005-01-01

    Full Text Available Earlier studies indicated that hormone responsiveness of cells and metabolic activity was lost during various of experimental procedure. In the light of this observation, I aimed to investigate to obtain optimal conditions for short time cultured hepatocytes and also to determine the type of test can be used to evaluate suitablity of hepatocytes for hormones studies. During the isolation period 50 IU/ml and 100 IU/ml collagenase were used. Adrenaline (10-6M was used to measure sensitivity of hepatocytes to hormones and glycogenolsis was measured at the end of 2hr incubation period. Adrenaline significantly increased gylcogenolysis (Control: 0.16±0.01 mg/2hr; Adrenaline: 0.30±0.01 mg/2hr only when the 50 IU/ml collagenase was used and the viability of the cells were over 95%. Viability tests were applied to hepatocytes that obtained by using 50 IU collagenase. Cellular glutathione, methylthiazoltetrazolium reduction, lactatedehdrogenase leakage, ATP level measured to determine viability following the attachment and incubation period. No differences were observed at the end of each period.Altogether, the present study indicated that membrane integrity and metabolic function of the hepatocytes can be improved by modifying slightly the original procedure of Reese and Byard.

  17. The nature and origin of Narrow Line AGN activity in a sample of isolated SDSS galaxies

    CERN Document Server

    Coziol, R; Plauchu-Frayn, I; Islas-Islas, J M; Ortega-Minakata, R A; Larios, D M Neri; Andernach, H

    2011-01-01

    We discuss the nature and origin of the nuclear activity observed in a sample of 292 SDSS narrow-emission-line galaxies, considered to have formed and evolved in isolation. All these galaxies are spiral like and show some kind of nuclear activity. The fraction of Narrow Line AGNs (NLAGNs) and Transition type Objects (TOs; a NLAGN with circumnuclear star formation) is relatively high, amounting to 64% of the galaxies. There is a definite trend for the NLAGNs to appear in early-type spirals, while the star forming galaxies and TOs are found in later-type spirals. We verify that the probability for a galaxy to show an AGN characteristic increases with the bulge mass of the galaxy (Torre-Papaqui et al. 2011), and find evidence that this trend is really a by-product of the morphology, suggesting that the AGN phenomenon is intimately connected with the formation process of the galaxies. Consistent with this interpretation, we establish a strong connection between the astration rate--the efficiency with which the ga...

  18. Origin of the excess IR luminosity in an apparently isolated LIRG

    Science.gov (United States)

    Cárdenas-Martínez, N.; Fuentes-Carrera, I.

    2016-06-01

    LIRGs and ULIRGs have long been recognized as one of the best laboratories to study the process of violent star formation in the Local Universe. In order for a star forming galaxy to emit at a LIRG level, it must have a very high star formation rate (SFR). In the local Universe, these high SFR are primarily triggered by galaxy-galaxy interactions or mergers. However, several authors show that roughly 50% of intermediate redshift LIRGs are disk galaxies without sign of recent interactions. The question arises regarding which processes could be originating such an enhanced SF. We present the extended kinematics of the ionized gas of an apparently isolated late-type galaxies displaying LIRG activity, as well as the morphological analysis from the UV to the MIR. We identify a morphological peculiarity which seems to be associated with double peaked profiles which could be related to two different kinematic components. We propose this to be a signature of an infalling satellite galaxy. CIG 302 might be an example of a minor interaction inducing very strong star formation in the local Universe.

  19. Mycobacterium avium subspecies impair dendritic cell maturation.

    Science.gov (United States)

    Basler, Tina; Brumshagen, Christina; Beineke, Andreas; Goethe, Ralph; Bäumer, Wolfgang

    2013-10-01

    Mycobacterium avium ssp. paratuberculosis (MAP) causes Johne's disease, a chronic, granulomatous enteritis of ruminants. Dendritic cells (DC) of the gut are ideally placed to combat invading mycobacteria; however, little is known about their interaction with MAP. Here, we investigated the interaction of MAP and the closely related M. avium ssp. avium (MAA) with murine DC and the effect of infected macrophages on DC maturation. The infection of DC with MAP or MAA induced DC maturation, which differed to that of LPS as maturation was accompanied by higher production of IL-10 and lower production of IL-12. Treatment of maturing DC with supernatants from mycobacteria-infected macrophages resulted in impaired DC maturation, leading to a semi-mature, tolerogenic DC phenotype expressing low levels of MHCII, CD86 and TNF-α after LPS stimulation. Though the cells were not completely differentiated they responded with an increased IL-10 and a decreased IL-12 production. Using recombinant cytokines we provide evidence that the semi-mature DC phenotype results from a combination of secreted cytokines and released antigenic mycobacterial components of the infected macrophage. Our results indicate that MAP and MAA are able to subvert DC function directly by infecting and indirectly via the milieu created by infected macrophages.

  20. Diversity of Virulence Factors Associated with West Australian Methicillin-Sensitive Staphylococcus aureus Isolates of Human Origin

    Directory of Open Access Journals (Sweden)

    Charlene Babra Waryah

    2016-01-01

    Full Text Available An extensive array of virulence factors associated with S. aureus has contributed significantly to its success as a major nosocomial pathogen in hospitals and community causing variety of infections in affected patients. Virulence factors include immune evading capsular polysaccharides, poly-N-acetyl glucosamine, and teichoic acid in addition to damaging toxins including hemolytic toxins, enterotoxins, cytotoxins, exfoliative toxin, and microbial surface components recognizing adhesive matrix molecules (MSCRAMM. In this investigation, 31 West Australian S. aureus isolates of human origin and 6 controls were analyzed for relative distribution of virulence-associated genes using PCR and/or an immunoassay kit and MSCRAMM by PCR-based typing. Genes encoding MSCRAMM, namely, Spa, ClfA, ClfB, SdrE, SdrD, IsdA, and IsdB, were detected in >90% of isolates. Gene encoding α-toxin was detected in >90% of isolates whereas genes encoding β-toxin and SEG were detectable in 50–60% of isolates. Genes encoding toxin proteins, namely, SEA, SEB, SEC, SED, SEE, SEH, SEI, SEJ, TSST, PVL, ETA, and ETB, were detectable in >50% of isolates. Use of RAPD-PCR for determining the virulence factor-based genetic relatedness among the isolates revealed five cluster groups confirming genetic diversity among the MSSA isolates, with the greatest majority of the clinical S. aureus (84% isolates clustering in group IIIa.

  1. [A case of environmental infection with pulmonary Mycobacterium avium complex disease from a residential bathroom of a patient suggested by variable-number tandem-repeat typing of Mycobacterium avium tandem repeat loci].

    Science.gov (United States)

    Taga, Shu; Niimi, Masaki; Kurokawa, Kazuhiro; Nakagawa, Taku; Ogawa, Kenji

    2012-05-01

    A 63-year-old woman was referred to our hospital because of bilateral infiltrations and nodular opacities in her chest radiograph taken in the mass radiography screening in September 2010. The chest computed tomography showed patchy infiltrations with bronchiectasis in the lower lung fields on both sides. She was diagnosed with pulmonary Mycobacterium avium complex (MAC) disease based on the bacteria recovered from the sputum and the bronchoalveolar lavage fluid. To elucidate an environmental MAC source, we investigated her home, and isolated M. avium and M. gordonae from the bathtub and shower tap, respectively, in her residential bathroom. Analysis of the hsp65-PRA variants digested with BamHI and some insertion sequences showed that the clinical strains recovered from sputum and strains from the bathtub were M. avium subsp. hominissuis. A dendrogram of the Mycobacterium avium tandem repeat loci variable-number tandem-repeat (MATR-VNTR) analysis of the MAC strains showed that the bathtub strains formed a polyclonal colonization, and that 1 of the 5 MATR-VNTR patterns was identical to the corresponding pattern of the sputum strain from the patient. In conclusion, we believe that the residential bathroom of the patient was the environmental source of her pulmonary MAC disease, as has been previously reported.

  2. Formulation of indomethacin emulsion using biopolymer of Prunus avium.

    Science.gov (United States)

    Verma, Shivangi; Dabral, Prashant; Rana, Vinod; Upadhaya, Kumud; Bhardwaj

    2012-03-01

    The aim of the investigation was to formulate Indomethacin Emulsion using Bio-polymer as Emulsifier. Different batches of emulsions were prepared by varying concentration of biopolymer prunus avium. Based evaluation of the prepared polymers, a conclusion can be drawn that in the Prunus avium bio-material can serve as a promising film forming agent for formulating various drug.

  3. Formulation of indomethacin emulsion using biopolymer of Prunus avium

    Directory of Open Access Journals (Sweden)

    Shivangi Verma

    2012-01-01

    Full Text Available The aim of the investigation was to formulate Indomethacin Emulsion using Bio-polymer as Emulsifier. Different batches of emulsions were prepared by varying concentration of biopolymer prunus avium. Based evaluation of the prepared polymers, a conclusion can be drawn that in the Prunus avium bio-material can serve as a promising film forming agent for formulating various drug.

  4. A Subinhibitory Concentration of Clarithromycin Inhibits Mycobacterium avium Biofilm Formation

    OpenAIRE

    2004-01-01

    Mycobacterium avium causes disseminated infection in immunosuppressed individuals and lung infection in patients with chronic lung diseases. M. avium forms biofilm in the environment and possibly in human airways. Antibiotics with activity against the bacterium could inhibit biofilm formation. Clarithromycin inhibits biofilm formation but has no activity against established biofilm.

  5. Comprehensive Insights in the Mycobacterium avium subsp. paratuberculosis Genome Using New WGS Data of Sheep Strain JIII-386 from Germany

    Science.gov (United States)

    Möbius, Petra; Hölzer, Martin; Felder, Marius; Nordsiek, Gabriele; Groth, Marco; Köhler, Heike; Reichwald, Kathrin; Platzer, Matthias; Marz, Manja

    2015-01-01

    Mycobacterium avium (M. a.) subsp. paratuberculosis (MAP)—the etiologic agent of Johne’s disease—affects cattle, sheep, and other ruminants worldwide. To decipher phenotypic differences among sheep and cattle strains (belonging to MAP-S [Type-I/III], respectively, MAP-C [Type-II]), comparative genome analysis needs data from diverse isolates originating from different geographic regions of the world. This study presents the so far best assembled genome of a MAP-S-strain: Sheep isolate JIII-386 from Germany. One newly sequenced cattle isolate (JII-1961, Germany), four published MAP strains of MAP-C and MAP-S from the United States and Australia, and M. a. subsp. hominissuis (MAH) strain 104 were used for assembly improvement and comparisons. All genomes were annotated by BacProt and results compared with NCBI (National Center for Biotechnology Information) annotation. Corresponding protein-coding sequences (CDSs) were detected, but also CDSs that were exclusively determined by either NCBI or BacProt. A new Shine–Dalgarno sequence motif (5′-AGCTGG-3′) was extracted. Novel CDSs including PE-PGRS family protein genes and about 80 noncoding RNAs exhibiting high sequence conservation are presented. Previously found genetic differences between MAP-types are partially revised. Four of ten assumed MAP-S-specific large sequence polymorphism regions (LSPSs) are still present in MAP-C strains; new LSPSs were identified. Independently of the regional origin of the strains, the number of individual CDSs and single nucleotide variants confirms the strong similarity of MAP-C strains and shows higher diversity among MAP-S strains. This study gives ambiguous results regarding the hypothesis that MAP-S is the evolutionary intermediate between MAH and MAP-C, but it clearly shows a higher similarity of MAP to MAH than to Mycobacterium intracellulare. PMID:26384038

  6. Illegitimate recombination: An efficient method for random mutagenesis in Mycobacterium avium subsp. hominissuis

    Directory of Open Access Journals (Sweden)

    Khattak Faisal

    2012-09-01

    Full Text Available Abstract Background The genus Mycobacterium (M. comprises highly pathogenic bacteria such as M. tuberculosis as well as environmental opportunistic bacteria called non-tuberculous mycobacteria (NTM. While the incidence of tuberculosis is declining in the developed world, infection rates by NTM are increasing. NTM are ubiquitous and have been isolated from soil, natural water sources, tap water, biofilms, aerosols, dust and sawdust. Lung infections as well as lymphadenitis are most often caused by M. avium subsp. hominissuis (MAH, which is considered to be among the clinically most important NTM. Only few virulence genes from M. avium have been defined among other things due to difficulties in generating M. avium mutants. More efforts in developing new methods for mutagenesis of M. avium and identification of virulence-associated genes are therefore needed. Results We developed a random mutagenesis method based on illegitimate recombination and integration of a Hygromycin-resistance marker. Screening for mutations possibly affecting virulence was performed by monitoring of pH resistance, colony morphology, cytokine induction in infected macrophages and intracellular persistence. Out of 50 randomly chosen Hygromycin-resistant colonies, four revealed to be affected in virulence-related traits. The mutated genes were MAV_4334 (nitroreductase family protein, MAV_5106 (phosphoenolpyruvate carboxykinase, MAV_1778 (GTP-binding protein LepA and MAV_3128 (lysyl-tRNA synthetase LysS. Conclusions We established a random mutagenesis method for MAH that can be easily carried out and combined it with a set of phenotypic screening methods for the identification of virulence-associated mutants. By this method, four new MAH genes were identified that may be involved in virulence.

  7. A 38-kilobase pathogenicity island specific for Mycobacterium avium subsp. paratuberculosis encodes cell surface proteins expressed in the host.

    Science.gov (United States)

    Stratmann, Janin; Strommenger, Birgit; Goethe, Ralph; Dohmann, Karen; Gerlach, Gerald-F; Stevenson, Karen; Li, Ling-Ling; Zhang, Qing; Kapur, Vivek; Bull, Tim J

    2004-03-01

    We have used representational difference analysis to identify a novel Mycobacterium avium subsp. paratuberculosis-specific ABC transporter operon (mpt), which comprises six open reading frames designated mptA to -F and is immediately preceded by two putative Fur boxes. Functional genomics revealed that the mpt operon is flanked on one end by a fep cluster encoding proteins involved in the uptake of Fe(3+) and on the other end by a sid cluster encoding non-ribosome-dependent heterocyclic siderophore synthases. Together these genes form a 38-kb M. avium subsp. paratuberculosis-specific locus flanked by an insertion sequence similar to IS1110. Expression studies using Western blot analyses showed that MptC is present in the envelope fraction of M. avium subsp. paratuberculosis. The MptD protein was shown to be surface exposed, using a specific phage (fMptD) isolated from a phage-peptide library, by differential screening of Mycobacterium smegmatis transformants. The phage fMptD-derived peptide could be used in a peptide-mediated capture PCR with milk from infected dairy herds, thereby showing surface-exposed expression of the MptD protein in the host. Together, these data suggest that the 38-kb locus constitutes an M. avium subsp. paratuberculosis pathogenicity island.

  8. Identification of Pasteurella multocida isolates of ruminant origin using polymerase chain reaction and their antibiogram study.

    Science.gov (United States)

    Kumar, P; Singh, V P; Agrawal, R K; Singh, S

    2009-04-01

    A total of 100 isolates of Pasteurella multocida from various ruminant species (cattle, buffalo and sheep) belonging to different parts of country were identified using Pasteurella multocida-PCR (PM-PCR) and capsular PCR assays. PM-PCR revealed an amplicon of approximately 460 bp in all the isolates tested. As regards capsular PCR, 36 of 38 cattle isolates and 30 of 34 buffalo isolates were found to belong to capsular serogroup B whereas rest of the cattle and buffalo isolates belonged to serogroup A of P. multocida. In case of sheep, a total of 26 out of 28 isolates were positive for serogroup A specific PCR while remaining 2 amplified a PCR product specific for serogroup F of P. multocida. All the isolates were subjected to antibiotic sensitivity testing using 17 different antibiotics. Enrofloxacin was found to be most potent antibiotic as it was effective against 94% of the isolates followed by ofloxacin (93%), chloramphenicol (93%), doxycycline (89%), tetracycline (86%) and ciprofloxacin (84%). Vancomycin, bacitracin and sulfadiazine were ineffective against P. multocida isolates showing 84%, 75% and 82% resistance, respectively. Further, the antibiogram also revealed the development of resistance against multiple drugs among various isolates of the organism.

  9. Prunus avium: nuclear DNA study in wild populations and sweet cherry cultivars.

    Science.gov (United States)

    Guarino, Carmine; Santoro, Simona; De Simone, Luciana; Cipriani, Guido

    2009-04-01

    The PCR-SSR technique was used to detect nuclear DNA diversity in five wild populations of Prunus avium from deciduous forests in Italy, Slovenia, and Croatia and 87 sweet cherry accessions from different geographical areas that have been maintained in the sweet cherry collection in Italy. This sweet cherry collection includes local accessions from the Campania Region as well as accessions from different countries. Twenty-eight microsatellites, previously developed in this species, generated polymorphic amplification products. Between 2 and 14 alleles were revealed for the polymorphic loci studied, with the expected heterozygosity ranging from 0.045 to 0.831. The total probability of identity was 56.94 x 10-18. A model-based Bayesian clustering analysis identified nine distinct gene pools in cultivated P. avium. The probability that wild populations were assigned to cultivated gene pools indicated that three gene pools accounted for the genomic origin of 53% of P. avium sampled. A dendrogram was generated using UPGMA (unweighted pair group method with arithmetic averages) based on Nei genetic distance analysis. This dendrogram classified most of the genotypes into one major group with an additional group of five accessions. The results indicate that this set of SSRs is highly informative, and they are discussed in terms of the implications for sweet cherry characterization.

  10. Genetic relationships between diploid and allotetraploid cherry species (Prunus avium, Prunus x gondouinii and Prunus cerasus).

    Science.gov (United States)

    Tavaud, M; Zanetto, A; David, J L; Laigret, F; Dirlewanger, E

    2004-12-01

    Prunus avium L. (diploid, AA, 2n=2x=16), Prunus cerasus L. (allotetraploid, AAFF, 2n=4x=32) species, and their hybrid Prunus x gondouinii Rehd., constitute the most widely cultivated cherry tree species. P. cerasus is supposed to be an hybrid species produced by the union of unreduced P. avium gametes and normal P. fruticosa gametes. A continuum of morphological traits between these three species makes their assignation difficult. The aim of this paper is to study the genetic relationships between tetraploid and diploid cherry species. In all, 114 genotypes belonging to these species were analyzed using 75 AFLP markers. The coordinates of these genotypes on the first axis of a correspondence analysis allowed us to clearly distinguish each species, to identify misclassifications and to assign unknown genotypes to one species. We showed that there are specific alleles in P. cerasus, which are not present in the A genome of P. avium and which probably come from the F genome of P. cerasus. The frequencies of each marker in the A and the F genomes were estimated in order to identify A and F specific markers. We discuss the utility of these specific markers for finding the origin of the A and F genomes in the allopolyploid species.

  11. Prevalence and diversity of enterotoxin genes with genetic background of Staphylococcus aureus isolates from different origins in China.

    Science.gov (United States)

    Chao, Guoxiang; Bao, Guangyu; Cao, Yongzhong; Yan, Wenguang; Wang, Yan; Zhang, Xiaorong; Zhou, Liping; Wu, Yantao

    2015-10-15

    Staphylococcal enterotoxins (SE) induce toxin-mediated diseases, such as food poisoning. In the present study, 568 isolates from different sources were tested for the prevalence of 18 SE genes and performed spa typing. In addition, we characterized the relationships between the distribution of SE genes and molecular clones based on multilocus sequence typing (MLST), spa and staphylococcal cassette chromosome mec (SCCmec) typing in selected 250 isolates. Approximately 54.40% of the isolates from different sources harbored one or more SE genes forming 120 distinct gene profiles. Seven genes, sea, seb, seg, seo, sem, seq, and sel were more frequently detected. The distributions of the SE genes among the isolates from human, animals, and foodborne origins were highly different with isolates from environments (P0.05). We identified two important gene clusters, sea-sek-seq, which is closely related to hospital-acquired (HA) methicillin-resistant Staphylococcus aureus (MRSA)-III, and the egc cluster, which accounts for nearly half of all genes. Approximately 71% isolates could be typed by spa, yielding 103 spa types, of which 18 spa types were primary types. In clonal complex (CC) 239, an important Asian HA-MRSA-III clone from humans, nearly all isolates harbored complete or partial sea-sek-seq cluster; the main spa types were t030 and t037. In CC630, an important new community-associated (CA) MRSA-V CC in China, only sporadic SE genes, three main spa types, t4549, t2196, and t377 were observed. The egc cluster coexisting with other genes was present in isolates of CC5, CC9, CC1281, CC1301, CC30 and sequence type (ST) 25, but completely absent in isolates of CC239, CC59, CC7, and CC88. The results illustrate the genetic clonal diversity and the identity of S. aureus isolates from different sources with respect to SE genes and highlight a correlation between SE genes or gene clusters and CCs, spa, and MRSA clones. The foodborne and human origin isolates were the main

  12. Direct identification and discernment of Mycobacterium avium and Mycobacterium intracellulare using a real-time RNA isothermal amplification and detection method.

    Science.gov (United States)

    Cui, Zhenling; Li, Yuanyuan; Cheng, Song; Yang, Hua; Lu, Junmei; Zhu, Honglei; Hu, Zhongyi

    2015-12-01

    The purpose of this work was to establish a real-time simultaneous amplification and testing method for identification and discernment of Mycobacterium avium and Mycobacterium intracellulare (SAT-MAC assay) and to evaluate the efficiency with which this method can detect isolated strains and clinical sputum specimens. The specific 16S rRNA sequences of M. avium and M. intracellulare were used as targets to design RNA probes and a reverse transcription primer containing T7 promoter. RNA isothermal amplification and real-time fluorescence detection were performed at 42 °C. SAT-MAC assay, culture tests on Lowenstein-Jensen (L-J) culture medium and PCR-sequencing were used to test the clinical isolated strains and sputum specimens. The limit of detection (LOD) of M. avium and M. intracellulare by SAT-MAC was found to be 30 CFU/mL and 20 CFU/mL. SAT-MAC showed high specificity in 21 species of mycobacteria standard strains and 5 species of non-mycobacteria bacteria. Using PCR-sequencing as the reference method, both rates of SAT-MAC assay for identifying M. avium and M. intracellulare from clinical isolates were 100% (259/259). Consistent with the results of L-J culture combined PCR-sequencing, the coincidence rate of SAT-MAC assay in clinical sputum specimens was 100% (369/369) for M. avium and 99.19% (366/369) for Mycobacterium intracellular. The SAT-MAC assay can identify and distinguish M. avium and M. intracellulare rapidly and accurately. It may be suitable for use in clinical microbiology laboratories.

  13. Population structure and diversity of the pathogenic fungus Aspergillus fumigatus isolated from different sources and geographic origins

    Directory of Open Access Journals (Sweden)

    Esperanza Duarte-Escalante

    2009-05-01

    Full Text Available Fifty-five clinical and environmental Aspergillus fumigatus isolates from Mexico, Argentina, France and Peru were analyzed to determine their genetic variability, reproductive system and level of differentiation using amplified fragment length polymorphism markers. The level of genetic variability was assessed by measuring the percentage of polymorphic loci, number of effective alleles, expected heterozygocity and by performing an association index test (I A. The degree of genetic differentiation and variation was determined using analysis of molecular variance at three levels. Using the paired genetic distances, a dendrogram was built to detect the genetic relationship among alleles. Finally, a network of haplotypes was constructed to determine the geographic relationship among them. The results indicate that the clinical isolates have greater genetic variability than the environmental isolates. The I A of the clinical and environmental isolates suggests a recombining population structure. The genetic differentiation among isolates and the dendrogram suggest that the groups of isolates are different. The network of haplotypes demonstrates that the majority of the isolates are grouped according to geographic origin.

  14. Effective heat inactivation of Mycobacterium avium subsp. paratuberculosis in raw milk contaminated with naturally infected feces.

    Science.gov (United States)

    Rademaker, Jan L W; Vissers, Marc M M; Te Giffel, Meike C

    2007-07-01

    The effectiveness of high-temperature, short holding time (HTST) pasteurization and homogenization with respect to inactivation of Mycobacterium avium subsp. paratuberculosis was evaluated quantitatively. This allowed a detailed determination of inactivation kinetics. High concentrations of feces from cows with clinical symptoms of Johne's disease were used to contaminate raw milk in order to realistically mimic possible incidents most closely. Final M. avium subsp. paratuberculosis concentrations varying from 10(2) to 3.5 x 10(5) cells per ml raw milk were used. Heat treatments including industrial HTST were simulated on a pilot scale with 22 different time-temperature combinations, including 60 to 90 degrees C at holding (mean residence) times of 6 to 15 s. Following 72 degrees C and a holding time of 6 s, 70 degrees C for 10 and 15 s, or under more stringent conditions, no viable M. avium subsp. paratuberculosis cells were recovered, resulting in >4.2- to >7.1-fold reductions, depending on the original inoculum concentrations. Inactivation kinetic modeling of 69 quantitative data points yielded an E(a) of 305,635 J/mol and an lnk(0) of 107.2, corresponding to a D value of 1.2 s at 72 degrees C and a Z value of 7.7 degrees C. Homogenization did not significantly affect the inactivation. The conclusion can be drawn that HTST pasteurization conditions equal to 15 s at > or =72 degrees C result in a more-than-sevenfold reduction of M. avium subsp. paratuberculosis.

  15. In Vitro Antimicrobial Susceptibility of Staphylococcus pseudintermedius Isolates of Human and Animal Origin

    Science.gov (United States)

    Wu, Max T.; Westblade, Lars F.; Robertson, Amy E.; Wallace, Meghan A.; Burd, Eileen M.; Hindler, Janet A.

    2016-01-01

    MIC results for 115 Staphylococcus intermedius group isolates are presented. Of these, 33% were methicillin resistant, among which 51.4% were susceptible to doxycycline, 29.7% to clindamycin, and 21.6% to trimethoprim-sulfamethoxazole. All of the isolates were susceptible to ceftaroline, daptomycin, linezolid, nitrofurantoin, quinupristin-dalfopristin, rifampin, tigecycline, and vancomycin. Of all the isolates, 82.6%, 67.8%, and 23.5% were susceptible to ciprofloxacin, erythromycin, and penicillin, respectively. No isolates harbored mupA or qacA/B genes, which suggested a lack of resistance to mupirocin or chlorhexidine. PMID:26962087

  16. Phenotypic characteristics of isolates of Aspergillus section Fumigati from different geographic origins and their relationships with genotypic characteristics

    Directory of Open Access Journals (Sweden)

    del Rocío Reyes-Montes María

    2011-05-01

    Full Text Available Abstract Background Epidemiological studies worldwide have shown that A. fumigatus exhibits important phenotypic and genotypic diversity, and these findings have been of great importance in improving the diagnosis and treatment of diseases caused by this fungus. However, few studies have been carried out related to the epidemiology of this fungus in Latin America. This study´s aim is to report on the epidemiology of the fungus by analyzing the phenotypic variability of Aspergillus section Fumigati isolates from different Latin American countries and the relationship between this variability, the geographical origin and genotypic characteristics. Methods We analyzed the phenotypic characteristics (macro- and micromorphology, conidial size, vesicles size, antifungal susceptibility and thermotolerance at 28, 37 and 48°C of A. section Fumigati isolates from Mexico (MX, Argentina (AR, Peru (PE and France (FR. The results were analyzed using analysis of variance (ANOVA and Tukey's multiple comparison test to detect significant differences. Two dendrograms among isolates were obtained with UPGMA using the Euclidean distance index. One was drawn for phenotypic data, and the other for phenotypic and genotypic data. A PCoA was done for shown isolates in a space of reduced dimensionality. In order to determine the degree of association between the phenotypic and genotypic characteristics AFLP, we calculated the correlation between parwise Euclidean distance matrices of both data sets with the nonparametric Mantel test. Results No variability was found in the macromorphology of the studied isolates; however, the micromorphology and growth rate showed that the PE isolates grew at a faster rate and exhibited the widest vesicles in comparison to the isolates from MX, AR and FR. The dendrogram constructed with phenotypic data showed three distinct groups. The group I and II were formed with isolates from PE and FR, respectively, while group III was formed

  17. Cooccurrence of free-living amoebae and nontuberculous Mycobacteria in hospital water networks, and preferential growth of Mycobacterium avium in Acanthamoeba lenticulata.

    Science.gov (United States)

    Ovrutsky, Alida R; Chan, Edward D; Kartalija, Marinka; Bai, Xiyuan; Jackson, Mary; Gibbs, Sara; Falkinham, Joseph O; Iseman, Michael D; Reynolds, Paul R; McDonnell, Gerald; Thomas, Vincent

    2013-05-01

    The incidence of lung and other diseases due to nontuberculous mycobacteria (NTM) is increasing. NTM sources include potable water, especially in households where NTM populate pipes, taps, and showerheads. NTM share habitats with free-living amoebae (FLA) and can grow in FLA as parasites or as endosymbionts. FLA containing NTM may form cysts that protect mycobacteria from disinfectants and antibiotics. We first assessed the presence of FLA and NTM in water and biofilm samples collected from a hospital, confirming the high prevalence of NTM and FLA in potable water systems, particularly in biofilms. Acanthamoeba spp. (genotype T4) were mainly recovered (8/17), followed by Hartmannella vermiformis (7/17) as well as one isolate closely related to the genus Flamella and one isolate only distantly related to previously described species. Concerning mycobacteria, Mycobacterium gordonae was the most frequently found isolate (9/17), followed by Mycobacterium peregrinum (4/17), Mycobacterium chelonae (2/17), Mycobacterium mucogenicum (1/17), and Mycobacterium avium (1/17). The propensity of Mycobacterium avium hospital isolate H87 and M. avium collection strain 104 to survive and replicate within various FLA was also evaluated, demonstrating survival of both strains in all amoebal species tested but high replication rates only in Acanthamoeba lenticulata. As A. lenticulata was frequently recovered from environmental samples, including drinking water samples, these results could have important consequences for the ecology of M. avium in drinking water networks and the epidemiology of disease due to this species.

  18. Separation of Salmonella typhimurium DT2 and DT135: Molecular characterization of isolates of avian origin

    DEFF Research Database (Denmark)

    Baggesen, Dorte Lau; Skov, M. N.; Brown, D. J.;

    1997-01-01

    characterization of 23 Danish isolates, 10 German isolates and the two type strains have subsequently been performed. With only minor exceptions, strains examined could be separated by combination of 0:5 agglutination, ribotyping, and PFGE typing into two major groups in conformity with their phage types...

  19. Clonal origin of aminoglycoside-resistant Citrobacter freundii isolates in a Danish county

    DEFF Research Database (Denmark)

    Norskov-Lauritsen, N.; Sandvang, Dorthe; Hedegaard, J.;

    2001-01-01

    During 1997, attention was drawn to an increased frequency of aminoglycoside-resistant Citrobacter freundii in a Danish county, when a total of 24 resistant C. freundii isolates was detected. In this study, 15 such isolates were typed by pulsed-field gel electrophoresis, riboprinting and partial...

  20. Neutralisation patterns among recent British and North American feline calicivirus isolates from different clinical origins.

    Science.gov (United States)

    Knowles, J O; Dawson, S; Gaskell, R M; Gaskell, C J; Harvey, C E

    1990-08-11

    The neutralisation patterns of 103 recent isolates of feline calicivirus from cats with chronic stomatitis or acute feline calicivirus disease, and from cats with neither oral nor respiratory disease were compared. There were no statistically significant differences between the proportions of isolates from each clinical source neutralised by individual feline calicivirus cat antisera. Different antisera showed widely differing degrees of cross reactivity; antisera to the most widely used vaccine strain F9 being the most cross reactive, neutralising 54 per cent of all the field isolates, and antisera to a field isolate LS015 the next most cross reactive, neutralising 29 per cent of the field isolates. However, the cross reactivity of antisera to early British isolates (A4, 68/40 and 69/1112) was much reduced (overall less than 10 per cent) whereas in the early 1970s 65 per cent of 117 field isolates from clinically normal cats were neutralised by A4 antiserum, and 40 per cent by each of 68/40 and 69/1112 antisera. This suggests a change in the spectrum of antigenicity among feline calicivirus isolates over the past 15 years. However, the cross reactivity of F9 antisera appeared to be similar to that in earlier studies. The relevance of these findings to vaccination is discussed.

  1. Activation of macrophages and interference with CD4+ T-cell stimulation by Mycobacterium avium subspecies paratuberculosis and Mycobacterium avium subspecies avium.

    Science.gov (United States)

    Zur Lage, Susanne; Goethe, Ralph; Darji, Ayub; Valentin-Weigand, Peter; Weiss, Siegfried

    2003-01-01

    Mycobacterium avium subspecies paratuberculosis (M. ptb) and M. avium subspecies avium (M. avium) are closely related but exhibit significant differences in their interaction with the host immune system. The macrophage line, J774, was infected with M. ptb and M. avium and analysed for cytokine production and stimulatory capacity towards antigen-specific CD4+ T cells. Under all conditions J774 cells were activated to produce proinflammatory cytokines. No influence on the expression of major histocompatibility complex (MHC) class II, intracellular adhesion molecule-1 (ICAM-1), B7.1, B7.2 or CD40 was found. However, the antigen-specific stimulatory capacity of J774 cells for a CD4+ T-cell line was significantly inhibited after infection with M. ptb, but not with M. avium. When a T-cell hybridoma expressing a T-cell receptor identical to that of the T-cell line was used, this inhibition was not observed, suggesting that costimulation which is essential for the CD4+ T-cell line is influenced by the pathogenic bacterium M. ptb.

  2. MYCOBACTERIUM AVIUM AS AN ACTUAL PATHOGEN OF HUMAN MYCOBACTERIOSIS

    Directory of Open Access Journals (Sweden)

    D. A. Starkova

    2013-01-01

    Full Text Available Abstract. Mycobacteriosis is an infectious disease of animals and humans caused by non-tuberculosis mycobacteria including M. avium complex. Despite the fact that the transmission of M. avium from human to human has not been proved, and mycobacteriosis has been sporadic, the number of cases of disseminated forms of disease caused by M. avium among HIV-positive patients during the last ten years was increasing. Limited knowledge about the structure of M. avium population in Russia and the lack of simple methods for the microbiological diagnosis make difficult the epidemiological monitoring of mycobacteriosis. This facilitates the use of modern, efficient molecular genetic methods for the species and subspecies identification and typing of M. avium. Thus, the detection of mobile element IS901, restriction fragment polymorphism analysis of hsp65 gene and IS1245 allow the detection and subspecies identification of M. avium. The study of genomic polymorphisms of bacterial strains for the assessment of M. avium population structure became feasible due to a complex of molecular techniques: VNTR-typing, IS1245-and IS1311-RFLP-typing.

  3. Gram-positive bacteria of marine origin: a numerical taxonomic study on Mediterranean isolates.

    Science.gov (United States)

    Ortigosa, M; Garay, E; Pujalte, M J

    1997-12-01

    A numerical taxonomic study was performed on 65 Gram-positive wild strains of heterotrophic, aerobic, marine bacteria, and 9 reference strains. The isolates were obtained from oysters and seawater sampled monthly over one year, by direct plating on Marine Agar. The strains were characterized by 96 morphological, biochemical, physiological and nutritional tests. Clustering yielded 13 phena at 0.62 similarity level (Sl coefficient). Only one of the seven phena containing wild isolates could be identified (Bacillus marinus). A pronounced salt requirement was found in most isolates.

  4. PRESENCE OF MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS IN ALPACAS (LAMA PACOS) INHABITING THE CHILEAN ALTIPLANO.

    Science.gov (United States)

    Salgado, Miguel; Sevilla, Iker; Rios, Carolina; Crossley, Jorge; Tejeda, Carlos; Manning, Elizabeth

    2016-03-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of paratuberculosis. The organism causes disease in both domestically managed and wild ruminant species. South American camelids have a long, shared history with indigenous people in the Andes. Over the last few decades, increasing numbers of alpacas were exported to numerous countries outside South America. No paratuberculosis surveillance has been reported for these source herds. In this study, individual fecal samples from 85 adult alpacas were collected from six separate herds in the Chilean Altiplano. A ParaTB mycobacterial growth indicator tube (MGIT) liquid culture of each individual fecal sample, followed by real-time polymerase chain reaction (PCR) protocol was used for confirmation. DNA extracts from a subset of confirmed MAP isolates were subjected to mycobacterial interspersed repetitive units-variable number of tandem repeats (MIRU-VNTR) typing. Fifteen alpaca were fecal culture test-positive. Five false-positive culture samples were negative on PCR analysis for Mycobacterium avium subsp. avium (MAA), Mycobacterium bovis (M. bovis), and the 16 S rDNA gene. Three MAP isolates subset-tested belonged to the same MIRU-VNTR type, showing four repeats for TR292 (locus 1) in contrast to the three repeats typical of the MAP reference strain K10. The number of repeats found in the remaining loci was identical to that of the K10 strain. It is not known how nor when MAP was introduced into the alpaca population in the Chilean Altiplano. The most plausible hypothesis to explain the presence of MAP in these indigenous populations is transmission by contact with infected domestic small ruminant species that may on occasion share pastures or range with alpacas. Isolation of this mycobacterial pathogen from such a remote region suggests that MAP has found its way beyond the confines of intensively managed domestic agriculture premises.

  5. Isolation, Identification, and Origin of Three Previously Unknown Congeners in Illicit Cocaine

    NARCIS (Netherlands)

    Ensing, Jacob; Hummelen, Jan C.

    1991-01-01

    Three previously unknown, overlooked, or perhaps wrongly identified impurities in illicit cocaine seized in the Netherlands Antilles are traced by various combinations of chromatographic and extraction methods. Once isolated using high-performance liquid chromatography, the compounds are identified

  6. Application of the C(18)-carboxypropylbetaine specimen processing method to recovery of Mycobacterium avium subsp. paratuberculosis from ruminant tissue specimens.

    Science.gov (United States)

    Thornton, Charles G; MacLellan, Kerry M; Stabel, Judith R; Carothers, Christine; Whitlock, Robert H; Passen, Selvin

    2002-05-01

    The causative agent of Johne's disease is Mycobacterium avium subsp. paratuberculosis. This is a chronic, debilitating gastrointestinal disorder that affects ruminants and is responsible for significant economic loss. The specimen processing method that combines C(18)-carboxypropylbetaine (CB-18) treatment and lytic enzyme decontamination has been shown to improve the diagnosis of mycobacterioses. This processing method was applied to the isolation of M. avium subsp. paratuberculosis from ruminant tissue samples. The BACTEC 12B liquid culture system was used but was supplemented with 1% egg yolk emulsion, 4 microg of mycobactin J, and 0.5% pyruvate (12B/EMP) for use in conjunction with this method. The final concentration of antibiotics used was 10 microg of vancomycin, 30 microg of amphotericin B, and 20 microg of nalidixic acid (VAN) per ml. A 7H10-based solid medium was also used that included mycobactin J, pyruvate, and VAN but excluded the egg yolk emulsion (7H10/MPV). Several M. avium subsp. paratuberculosis isolates were examined during the evaluation of this processing method. It was observed that treatment with lytic enzymes stimulated the growth of M. avium subsp. paratuberculosis; however, the growth of one isolate was markedly inhibited due to the presence of vancomycin. Subsequently, the vancomycin concentration in the VAN formulation was reduced to 2 microg/ml. A blinded panel of 60 previously characterized tissue samples from bovine and bison were then processed and analyzed by smear and culture. Historically, 31 and 37 specimens were classified as positive by histology and culture, respectively. The overall sensitivity and specificity of smear relative to culture following CB-18 processing were 97.6 and 89.5%, respectively. The 12B/EMP/VAN liquid culture system recovered M. avium subsp. paratuberculosis from 39 specimens, whereas 7H10/MPV and Herrold's egg yolk media recovered M. avium subsp. paratuberculosis from 26 and 16 specimens, respectively

  7. The Origin of Isolated Early-Type Galaxies: A Multiwavelength Study of Three Systems

    Science.gov (United States)

    Fanelli, Michael N.; Marcum, Pamela M.; Ashley, Trisha L.; Fuse, Christopher R.; O'Toole Appleby, Heather

    2017-01-01

    One major approach to understanding the zero-redshift galaxy population is to distinguish the processes driving passive (internal) evolution of stellar systems from interaction-induced changes. The study of the structure and star formation history of highly isolated systems defines the baseline for quiescent evolution. Isolated early-type galaxies (ETGs) are an especially interesting population, since most ETGs are found in cluster environments. Possible evolutionary paths for isolated ETGs are merged (formerly) compact groups or direct collapse early in cosmic time.As part of our long-term program to identify and explore highly isolated ETGs, we present panchromatic observations of the stars, neutral gas and dust in three highly isolated systems: Mrk 150, KIG 824 and SDSS J235021+141343, a subset of our initial isolated ETG sample. Our goal is to understand the evolutionary paths for these systems, in terms of their merger history, gas fueling and star formation rates. Each system presents diverse properties: Mrk 150 is optically blue and currently forming stars, while KIG 824 is red with no evidence for ongoing star formation; all possess neutral gas. Mrk 150 fits the class of luminous blue compact galaxies [M(V) = -19.2, B-V = 0.47] and possibly represents a link in the evolutionary chain leading to isolated, red and dead systems.We combine GALEX UV photometry, optical images and spectra from the Sloan Survey, IR photometry from 2MASS, WISE and IRAS along with neutral hydrogen observations from the GBT. These data permit us to describe their morphology, quantify the current high-mass star formation rate, and determine the gas and dust content. Derived properties help constrain the evolution of these particular systems in the larger context of ETGs evolving in void-like regions.

  8. Draft Genome Sequences of Six Ruminant Coxiella burnetii Isolates of European Origin

    DEFF Research Database (Denmark)

    Sidi-Boumedine, Karim; Ellis, Richard J.; Adam, Gilbert

    2014-01-01

    Coxiella burnetii is responsible for Q fever, a worldwide zoonosis attributed to the inhalation of aerosols contaminated by livestock birth products. Six draft genome sequences of European C. burnetii isolates from ruminants are presented here. The availability of these genomes will help in under...

  9. Pathogenicity of Phytophthora isolates originating from several woody hosts in Bulgaria and Poland

    Directory of Open Access Journals (Sweden)

    Lyubenova Aneta B.

    2016-09-01

    Full Text Available Our aim was to examine the virulence of eight Phytophthora isolates belonging to three species (Phytophthora cryptogea, Phytophthora plurivora and Phytophthora quercina obtained from diverse European ecosystems (in Bulgaria, Poland and Germany towards three forest tree hosts – English oak (Quercus robur L., Turkey oak (Quercus cerris L. and European beech (Fagus sylvatica L..

  10. Surveillance of Mycobacterium avium subsp. paratuberculosis in dairy herds

    NARCIS (Netherlands)

    Weber, M.F.

    2009-01-01

    In this thesis, the potential for improvements in surveillance of Mycobacterium avium subsp. paratuberculosis (Map) infection and paratuberculosis in dairy herds was investigated, leading to a reduction in surveillance costs whilst continuing to meet specific quality targets. In particular, differen

  11. The origin of the isolated population of the Faroe Islands investigated using Y chromosomal markers

    DEFF Research Database (Denmark)

    Jorgensen, Tove H; Buttenschön, Henriette N; Wang, August G;

    2004-01-01

    and the Norwegian, Swedish and Icelandic Y chromosomes but also some similarity with the Scottish and Irish Y chromosomes. Diversity measures and estimates of effective population sizes also suggest that the original gene pool of the settlers have been influenced by random genetic drift, thus complicating direct...

  12. Isolamento de cepas de Mycobacterium avium em búfalos abatidos para consumo Mycobacterium avium complex in water buffaloes slaughtered for consumption

    Directory of Open Access Journals (Sweden)

    José de Arimatéa Freitas

    2001-06-01

    Full Text Available Duas cepas micobacterianas, isoladas no parênquima pulmonar e linfonodo apical de búfalos abatidos para consumo, procedentes de criatórios localizados na Ilha de Marajó (PA e submetidas à identificação segundo ensaios recomendados para o gênero Mycobacterium, foram identificadas como pertencentes ao complexo Mycobacterium avium. Apresentaram-se considerações relativas à associação desses organismos com a Aids -- e o papel dos alimentos nessa associação --, discutindo-se o impacto que a condição de germes oportunistas das espécies desse complexo têm na pandemia do HIV, assim como o risco potencial representado pelas infecções produzidas nos animais.Two mycobacterium strains isolated from lung tissue and apical lymph nodes of slaughtered water buffaloes were biochemically analyzed and identified as Mycobacterium avium complex strains. Association between these microorganisms and the acquired immunodeficiency syndrome, and the potential risk posed by eating infected animals and their products, was discussed.

  13. Prevalence of quinolone resistance determinants in non-typhoidal Salmonella isolates from human origin in Extremadura, Spain.

    Science.gov (United States)

    Campos, Maria Jorge; Palomo, Gonzalo; Hormeño, Lorena; Herrera-León, Silvia; Domínguez, Lucas; Vadillo, Santiago; Píriz, Segundo; Quesada, Alberto

    2014-05-01

    Resistance to the quinolones nalidixic acid (NAL) and ciprofloxacin (CIP) and the occurrence of quinolone resistance determinants have been investigated in 300 non-typhoidal Salmonella from human origin, isolated in the years between 2004 and 2008, in 6 hospitals within Extremadura (Spain). Salmonella Enteritidis was the major serotype found among quinolone-resistant isolates, most of which were clustered by clonal analysis to a single clone, which presented D87 or S83 substitutions in GyrA. Eleven isolates presented the non-classical quinolone resistance phenotype (resistance to CIP and susceptibility to NAL), lacking mutations in the quinolone resistance determinant region of topoisomerase genes. Among them, one Salmonella Typhimurium isolate carried a qnrS1 gene in a low-molecular-weight plasmid, pQnrS1-HLR25, identical to plasmids previously found in the UK, Taiwan, and USA. The occurrence of this genetic element could represent a risk for the horizontal transmission of quinolone resistance among Enterobacteriaceae in the Iberian Peninsula.

  14. Human Origin Lactobacillus casei Isolated from Indonesian Infants Demonstrating Potential Characteristics as Probiotics in vitro

    Directory of Open Access Journals (Sweden)

    Widodo .

    2015-11-01

    Full Text Available Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} The aim of this experiment was to isolate and identify Lactic Acid Bacteria (LAB from infant faeces and subsequent evaluation of its potential probiotics. LAB was isolated from faeces of infants who consumed breast milk as the only source of diet on L-cysteine-supplemented MRS Agar, and incubated on 37oC for 48 hours. Colonies grew on this media were then identifi ed based on morphological, physiological and molecular approaches. Morphological and physiological identifi cations based on Gram staining, shape, motility, spore formation, catalase, CO2 and NH3 production, and the ability to grow on temperature at 10oC and 45oC. Molecular identifi cation based on the amplifi cation of 16S rRNA gene. The potential application of selected isolates for probiotics was evaluated based on the ability to grow on media with low pH and the addition of 0.5% bile salts, the ability to inhibit the growth of pathogenic Bacillus cereus and Eschericia coli, and in vitroadherence ability. On the basis of morphological, physiological and molecular analysis of 16S rRNA gene, it was concluded that the selected isolate 1AF was a strain of Lactobacillus casei. Evaluation of probiotic in vitro showed that 60.4% of cells were resistant

  15. Evidence for the existence of two new members of the family Chlamydiaceae and proposal of Chlamydia avium sp. nov. and Chlamydia gallinacea sp. nov.

    Science.gov (United States)

    Sachse, Konrad; Laroucau, Karine; Riege, Konstantin; Wehner, Stefanie; Dilcher, Meik; Creasy, Heather Huot; Weidmann, Manfred; Myers, Garry; Vorimore, Fabien; Vicari, Nadia; Magnino, Simone; Liebler-Tenorio, Elisabeth; Ruettger, Anke; Bavoil, Patrik M; Hufert, Frank T; Rosselló-Móra, Ramon; Marz, Manja

    2014-03-01

    The family Chlamydiaceae with the recombined single genus Chlamydia currently comprises nine species, all of which are obligate intracellular organisms distinguished by a unique biphasic developmental cycle. Anecdotal evidence from epidemiological surveys in flocks of poultry, pigeons and psittacine birds have indicated the presence of non-classified chlamydial strains, some of which may act as pathogens. In the present study, phylogenetic analysis of ribosomal RNA and ompA genes, as well as multi-locus sequence analysis of 11 field isolates were conducted. All independent analyses assigned the strains into two different clades of monophyletic origin corresponding to pigeon and psittacine strains or poultry isolates, respectively. Comparative genome analysis involving the type strains of currently accepted Chlamydiaceae species and the designated type strains representing the two new clades confirmed that the latter could be classified into two different species as their average nucleotide identity (ANI) values were always below 94%, both with the closest relative species and between themselves. In view of the evidence obtained from the analyses, we propose the addition of two new species to the current classification: Chlamydia avium sp. nov. comprising strains from pigeons and psittacine birds (type strain 10DC88(T); DSMZ: DSM27005(T), CSUR: P3508(T)) and Chlamydia gallinacea sp. nov. comprising strains from poultry (type strain 08-1274/3(T); DSMZ: DSM27451(T), CSUR: P3509(T)).

  16. Detection of mycobacteria, Mycobacterium avium subspecies, and Mycobacterium tuberculosis complex by a novel tetraplex real-time PCR assay.

    Science.gov (United States)

    Sevilla, Iker A; Molina, Elena; Elguezabal, Natalia; Pérez, Valentín; Garrido, Joseba M; Juste, Ramón A

    2015-03-01

    Mycobacterium tuberculosis complex, Mycobacterium avium, and many other nontuberculous mycobacteria are worldwide distributed microorganisms of major medical and veterinary importance. Considering the growing epidemiologic significance of wildlife-livestock-human interrelation, developing rapid detection tools of high specificity and sensitivity is vital to assess their presence and accelerate the process of diagnosing mycobacteriosis. Here we describe the development and evaluation of a novel tetraplex real-time PCR for simultaneous detection of Mycobacterium genus, M. avium subspecies, and M. tuberculosis complex in an internally monitored single assay. The method was evaluated using DNA from mycobacterial (n = 38) and nonmycobacterial (n = 28) strains, tissues spiked with different CFU amounts of three mycobacterial species (n = 57), archival clinical samples (n = 233), and strains isolated from various hosts (n = 147). The minimum detectable DNA amount per reaction was 50 fg for M. bovis BCG and M. kansasii and 5 fg for M. avium subsp. hominissuis. When spiked samples were analyzed, the method consistently detected as few as 100 to 1,000 mycobacterial CFU per gram. The sensitivity and specificity values for the panel of clinical samples were 97.5 and 100% using a verified culture-based method as the reference method. The assays performed on clinical isolates confirmed these results. This PCR was able to identify M. avium and M. tuberculosis complex in the same sample in one reaction. In conclusion, the tetraplex real-time PCR we designed represents a highly specific and sensitive tool for the detection and identification of mycobacteria in routine laboratory diagnosis with potential additional uses.

  17. Recent advances in isolation, synthesis, and evaluation of bioactivities of bispyrroloquinone alkaloids of marine origin.

    Science.gov (United States)

    Nijampatnam, Bhavitavya; Dutta, Shilpa; Velu, Sadanandan E

    2015-08-01

    The ocean continues to provide a plethora of unique scaffolds capable of remarkable biological applications. A large number of pyrroloiminoquinone alkaloids, including discorhabdins, epinardins, batzellines, makaluvamines, and veiutamine, have been isolated from various marine organisms. A class of pyrroloiminoquinone-related alkaloids, known as bispyrroloquinones, is the focus of this review article. This family of marine alkaloids, which contain an aryl substituted bispyrroloquinone ring system, includes three subclasses of alkaloids namely, wakayin, tsitsikammamines A-B, and zyzzyanones A-D. Both wakayin and the tsitsikammamines contain a tetracyclic fused bispyrroloiminoquinone ring system, while zyzzyanones contain a fused tricyclic bispyrroloquinone ring system. The unique chemical structures of these marine natural products and their diverse biological properties, including antifungal and antimicrobial activity, as well as the potent, albeit generally nonspecific and universal cytotoxicities, have attracted great interest of synthetic chemists over the past three decades. Tsitsikammamines, wakayin, and several of their analogs show inhibition of topoisomerases. One additional possible mechanism of anticancer activity of tsitsikammamines analogs that has been discovered recently is through the inhibition of indoleamine 2, 3-dioxygenase, an enzyme involved in tumoral immune resistance. This review discusses the isolation, synthesis, and evaluation of bioactivities of bispyrroloquinone alkaloids and their analogs.

  18. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

    Science.gov (United States)

    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-12-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent.

  19. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

    Science.gov (United States)

    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-01-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent. Images PMID:6094355

  20. The origin of the isolated population of the Faroe Islands investigated using Y chromosomal markers

    DEFF Research Database (Denmark)

    Jorgensen, Tove H; Buttenschön, Henriette N; Wang, August G;

    2004-01-01

    and the Norwegian, Swedish and Icelandic Y chromosomes but also some similarity with the Scottish and Irish Y chromosomes. Diversity measures and estimates of effective population sizes also suggest that the original gene pool of the settlers have been influenced by random genetic drift, thus complicating direct...... to analyse genetic diversity in the Faroese population and to compare this with the distribution of genotypes in the putative ancestral populations. Using a combination of genetic distance measures, assignment and phylogenetic analyses, we find a high degree of similarity between the Faroese Y chromosomes...

  1. Characterization of Biofilm Formation in [Pasteurella] pneumotropica and [Actinobacillus] muris Isolates of Mouse Origin.

    Science.gov (United States)

    Sager, Martin; Benten, W Peter M; Engelhardt, Eva; Gougoula, Christina; Benga, Laurentiu

    2015-01-01

    [Pasteurella] pneumotropica biotypes Jawetz and Heyl and [Actinobacillus] muris are the most prevalent Pasteurellaceae species isolated from laboratory mouse. However, mechanisms contributing to their high prevalence such as the ability to form biofilms have not been studied yet. In the present investigation we analyze if these bacterial species can produce biofilms in vitro and investigate whether proteins, extracellular DNA and polysaccharides are involved in the biofilm formation and structure by inhibition and dispersal assays using proteinase K, DNase I and sodium periodate. Finally, the capacity of the biofilms to confer resistance to antibiotics is examined. We demonstrate that both [P.] pneumotropica biotypes but not [A.] muris are able to form robust biofilms in vitro, a phenotype which is widely spread among the field isolates. The biofilm inhibition and dispersal assays by proteinase and DNase lead to a strong inhibition in biofilm formation when added at the initiation of the biofilm formation and dispersed pre-formed [P.] pneumotropica biofilms, revealing thus that proteins and extracellular DNA are essential in biofilm formation and structure. Sodium periodate inhibited the bacterial growth when added at the beginning of the biofilm formation assay, making difficult the assessment of the role of β-1,6-linked polysaccharides in the biofilm formation, and had a biofilm stimulating effect when added on pre-established mature biofilms of [P.] pneumotropica biotype Heyl and a majority of [P.] pneumotropica biotype Jawetz strains, suggesting that the presence of β-1,6-linked polysaccharides on the bacterial surface might attenuate the biofilm production. Conversely, no effect or a decrease in the biofilm quantity was observed by biofilm dispersal using sodium periodate on further biotype Jawetz isolates, suggesting that polysaccharides might be incorporated in the biofilm structure. We additionally show that [P.] pneumotropica cells enclosed in biofilms

  2. Multilocus sequence typing reveals that Bacillus cereus strains isolated from clinical infections have distinct phylogenetic origins.

    Science.gov (United States)

    Barker, Margaret; Thakker, Bishan; Priest, Fergus G

    2005-04-01

    Eight strains of Bacillus cereus isolated from bacteremia and soft tissue infections were assigned to seven sequence types (STs) by multilocus sequence typing (MLST). Two strains from different locations had identical STs. The concatenated sequences of the seven STs were aligned with 65 concatenated sequences from reference STs and a neighbor-joining tree was constructed. Two strains were distantly related to all reference STs. Three strains were recovered in a clade that included Bacillus anthracis, B. cereus and rare Bacillus thuringiensis strains while the other three strains were assigned to two STs that were more closely affiliated to most of the B. thuringiensis STs. We conclude that invasive B. cereus strains do not form a single clone or clonal complex of highly virulent strains.

  3. In vitro antibiotic susceptibility of Dutch Mycoplasma synoviae field isolates originating from joint lesions and the respiratory tract of commercial poultry

    NARCIS (Netherlands)

    Landman, W.J.M.; Mevius, D.J.; Veldman, K.T.; Feberwee, A.

    2008-01-01

    The in vitro susceptibility of 17 Dutch Mycoplasma synoviae isolates from commercial poultry to enrofloxacin, difloxacin, doxycycline, tylosin and tilmicosin was examined. Three isolates originated from joint lesions and 14 were from the respiratory tract. The type strain M. synoviae WVU 1853 was in

  4. Detection of a 640-bp deletion in the Aggregatibacter actinomycetemcomitans leukotoxin promoter region in isolates from an adolescent of Ethiopian origin

    Directory of Open Access Journals (Sweden)

    Rolf Claesson

    2015-04-01

    Full Text Available The expression of the leukotoxin of Aggregatibacter actinomycetemcomitans is regulated by the leukotoxin promoter. A 530-bp deletion or an 886-bp insertion sequence (IS element in this region has earlier been described in highly leukotoxic isolates. Here, we report on highly leukotoxic isolate with a 640-bp deletion, which was detected in an adolescent of Ethiopian origin.

  5. Biosynthetic origin of natural products isolated from marine microorganism-invertebrate assemblages.

    Science.gov (United States)

    Simmons, T Luke; Coates, R Cameron; Clark, Benjamin R; Engene, Niclas; Gonzalez, David; Esquenazi, Eduardo; Dorrestein, Pieter C; Gerwick, William H

    2008-03-25

    In all probability, natural selection began as ancient marine microorganisms were required to compete for limited resources. These pressures resulted in the evolution of diverse genetically encoded small molecules with a variety of ecological and metabolic roles. Remarkably, many of these same biologically active molecules have potential utility in modern medicine and biomedical research. The most promising of these natural products often derive from organisms richly populated by associated microorganisms (e.g., marine sponges and ascidians), and often there is great uncertainty about which organism in these assemblages is making these intriguing metabolites. To use the molecular machinery responsible for the biosynthesis of potential drug-lead natural products, new tools must be applied to delineate their genetic and enzymatic origins. The aim of this perspective is to highlight both traditional and emerging techniques for the localization of metabolic pathways within complex marine environments. Examples are given from the literature as well as recent proof-of-concept experiments from the authors' laboratories.

  6. Molecular characterization of Spanish Prunus avium plus trees

    Directory of Open Access Journals (Sweden)

    Javier Fernandez-Cruz

    2014-04-01

    Full Text Available Aim of the study: The Breeding Program of wild cherry (Prunus avium developed by Lourizán Forest Research Center (NW Spain, aims for the creation of the Main Breeding Population, that is formed by a large number of plus trees and for obtaining an Elite Population generated from controlled crosses of a number of plus trees selected by, at least, one trait of economic importance. The aim of this study was to genotype 131 accessions of Prunus avium plus trees, included in the breeding program.Area of study: The Prunus avium plus trees are located in the North, Northwest and Central Spain.Material and Methods: The Prunus avium plus trees were genotyped with nine microsatellites. Several genetic parameters were calculated. Genetic data were analyzed with STRUCTURE and the genetic distance between the plus trees were calculated.Main results: A total of 122 multilocus genotypes were detected. Several accessions with the same genotype were identified, which could be due to clonality or to labelling errors. The nine microsatellites are useful for identifying individuals because the combined probability of identity was low (PI = 5.19X10-9. Bayesian methods detected two genetic clusters in the sampled plus trees.Research highlights: The unique genotypes identified in this work are suitable for being included in the elite breeding population for economic traits.Keywords: Prunus avium; breeding program; microsatellite; genetic distance.

  7. A cDNA encoding a cold-induced glycine-rich RNA binding protein from Prunus avium expressed in embryonic axes.

    Science.gov (United States)

    Stephen, John R; Dent, Katherine C; Finch-Savage, William E

    2003-11-27

    A cDNA clone encoding a presumed full-length glycine-rich ribonucleic acid (RNA) binding protein was isolated from a lambda-ZAP Express cDNA library generated from primarily nondormant Prunus avium (wild cherry) embryonic axes. The cDNA, designated Pa-RRM-GRP1 (Prunus avium RNA recognition motif glycine-rich protein 1), contains a single N-terminal RNA recognition motif (RRM) and single C-terminal glycine-rich domain. The glycine-rich domain is unusually long at 91 amino acids, 58 of which are glycines. The 534-base pair (bp) open reading frame (ORF) of this clone encodes a 178-amino-acid polypeptide with a predicted molecular weight of 17.33 kDa and pI of 7.84. Comparative sequence alignment of Pa-RRM-GRP1 reveals extensive homology to known and presumed glycine-rich RNA binding proteins from angiosperms and gymnosperms. Genomic Southern blot analysis suggests that this gene exists as a single copy in P. avium. Expression of this gene in P. avium embryonic axes during low-temperature dormancy-breaking treatments was studied and found to be induced by cold (3 degrees C) using real-time PCR of total cDNA supported by Northern blot analysis of total RNA. Expression dropped during prolonged storage at 3 degrees C and was reduced to control levels by interruption of cold treatment by warming to 20 degrees C.

  8. Exploring the zoonotic potential of Mycobacterium avium subspecies paratuberculosis through comparative genomics.

    Directory of Open Access Journals (Sweden)

    James W Wynne

    Full Text Available A comparative genomics approach was utilised to compare the genomes of Mycobacterium avium subspecies paratuberculosis (MAP isolated from early onset paediatric Crohn's disease (CD patients as well as Johne's diseased animals. Draft genome sequences were produced for MAP isolates derived from four CD patients, one ulcerative colitis (UC patient, and two non-inflammatory bowel disease (IBD control individuals using Illumina sequencing, complemented by comparative genome hybridisation (CGH. MAP isolates derived from two bovine and one ovine host were also subjected to whole genome sequencing and CGH. All seven human derived MAP isolates were highly genetically similar and clustered together with one bovine type isolate following phylogenetic analysis. Three other sequenced isolates (including the reference bovine derived isolate K10 were genetically distinct. The human isolates contained two large tandem duplications, the organisations of which were confirmed by PCR. Designated vGI-17 and vGI-18 these duplications spanned 63 and 109 open reading frames, respectively. PCR screening of over 30 additional MAP isolates (3 human derived, 27 animal derived and one environmental isolate confirmed that vGI-17 and vGI-18 are common across many isolates. Quantitative real-time PCR of vGI-17 demonstrated that the proportion of cells containing the vGI-17 duplication varied between 0.01 to 15% amongst isolates with human isolates containing a higher proportion of vGI-17 compared to most animal isolates. These findings suggest these duplications are transient genomic rearrangements. We hypothesise that the over-representation of vGI-17 in human derived MAP strains may enhance their ability to infect or persist within a human host by increasing genome redundancy and conferring crude regulation of protein expression across biologically important regions.

  9. Effects of polysaccharide on chicks co-infected with Bordetella avium and Avian leukosis virus.

    Science.gov (United States)

    Guo, Fanxia; Xue, Cong; Wu, Cun; Zhao, Xue; Qu, Tinghe; He, Xiaohua; Guo, Zhongkun; Zhu, Ruiliang

    2014-08-30

    Chicks' co-infection with immunosuppressive virus and bacteria seriously threaten the development of the poultry industry. In this study, a model was established in which chicks were injected with either subgroup B ALV (ALV-B)+Bordetella avium (B. avium), or ALV-B+B. avium+Taishan Pinus massoniana pollen polysaccharide (TPPPS), or B. avium only, or B. avium+TPPPS. The data showed that the group injected with ALV-B and B. avium exhibited significant inhibition of the immune function and therefore increased pathogenicity compared with the group injected with B. avium-only. Application of TPPPS effectively alleviated immunosuppression, and body weights increased sharply in the TPPPS groups compared with non-TPPPS groups. To some extent, TPPPS may reduce the proliferation of ALV-B. These results suggest that Pinus pollen polysaccharides are beneficial treating co-infections with immunosuppressive virus and bacteria and therefore have potential for development into safe and effective immunoregulator.

  10. Discrimination and divergence among Lactobacillus plantarum-group (LPG) isolates with reference to their probiotic functionalities from vegetable origin.

    Science.gov (United States)

    Devi, Sundru Manjulata; Aishwarya, Subramanian; Halami, Prakash M

    2016-12-01

    The present study was aimed to evaluate the diversity and probiotic properties of Lactobacillus plantarum-group cultures from vegetable origin. First, genotypic diversity of L. plantarum (n=34) was achieved by PCR of Random Amplified Polymorphic DNA and recA gene-specific multiplex PCR. The isolates were segregated into five groups namely, Lactobacillus pentosus, Lactobacillus paraplantarum, Lactobacillus arizonensis, Lactobacillus plantarum subsp. plantarum and argentoratensis. Further discrimination was achieved by restriction fragment length polymorphism of probiotic adhesion genes viz.fbp, mub and msa gene. As determined by nucleotide sequence analysis and bioinformatics Pfam database, the putative Fbp protein had only one FBP domain, whereas Mub protein had 8-10 MUB domain repeats. However, L. pentosus (except CFR MFT9), L. plantarum subsp. argentoratensis (except CFR MFT5) and L. arizonensis (except CFR MFT2) isolates gave no amplicon for the tested marker genes. Selected cultures (n=15) showed tolerance to simulated digestive fluids (20-85%), exhibited auto-aggregation (10-77%), cellular hydrophobicity (12-78%), and broad spectrum of anti-microbial activity. Concurrently, high adherence capacity to mucin was achieved for L. plantarum subsp. plantarum (MCC 2974 and CFR MFT1) and L. paraplantarum (MTCC 9483, MCC 2977, MCC 2978), which had an additional MUB domain repeat.

  11. Experimental infection of Eurasian wild boar with Mycobacterium avium subsp. avium.

    Science.gov (United States)

    Garrido, J M; Vicente, J; Carrasco-García, R; Galindo, R C; Minguijón, E; Ballesteros, C; Aranaz, A; Romero, B; Sevilla, I; Juste, R; de la Fuente, J; Gortazar, C

    2010-07-29

    The Eurasian wild boar (Sus scrofa) is increasingly relevant as a host for several pathogenic mycobacteria. We aimed to characterize the first experimental Mycobacterium avium subsp. avium (MAA) infection in wild boar in order to describe the lesions and the immune response as compared to uninfected controls. Twelve 1-4-month-old wild boar piglets were housed in class III bio-containment facilities. Four concentrations of MAA suspension were used: 10, 10(2) and 10(4) mycobacteria (2 animals each, oropharyngeal route) and 2.5 x 10(6) mycobacteria (2 animals each by the oropharyngeal and nasal routes). No clinical signs were observed and pathology evidenced a low pathogenicity of this MAA strain for this particular host. Bacteriological and pathological evidence of successful infection after experimental inoculation was found for the group challenged with 2.5 x 10(6) mycobacteria. These four wild boar showed a positive IFN-gamma response to the avian PPD and the real-time RT-PCR data revealed that three genes, complement component C3, IFN-gamma and RANTES, were significantly down regulated in infected animals. These results were similar to those found in naturally and experimentally M. bovis-infected wild boar and may constitute biomarkers of mycobacterial infection in this species.

  12. Impact of the origin of sinus node artery on recurrence after pulmonary vein isolation in patients with paroxysmal atrial fibrillation

    Institute of Scientific and Technical Information of China (English)

    ZHANG Zhi-jun; CHEN Ke; TANG Ri-bo; gANG Cai-hua; Edmundo Patricio Lopes Lao; YAN Qian; HE Xiao-nan

    2013-01-01

    Background Major atrial coronary arteries,including the sinus node artery (SNA),were commonly found in the areas involved in atrial fibrillation (AF) ablation and could cause difficulties in achieving linear block at the left atrial (LA) roof.The SNA is a major atrial coronary artery of the atrial coronary circulation.This study aimed to determine impact of the origin of SNA on recurrence of AF after pulmonary vein isolation (PVI) in patients with paroxysmal AF.Methods Seventy-eight patients underwent coronary angiography for suspected coronary heart disease,followed by catheter ablation for paroxysmal AF.According to the origin of SNA from angiographic findings,they were divided into right SNA group (SNA originating from the right coronary artery) and left SNA group (SNA originating from the left circumflex artery).Guided by an electroanatomic mapping system,circumferential pulmonary vein ablation (CPVA) was performed in both groups and PVI was the procedural endpoint.All patients were followed up at 1,3,6,9 and 12 months post-ablation.Recurrence was defined as any episode of atrial tachyarrhythmias (ATAs),including AF,atrial flutter or atrial tachycardia,that lasted longer than 30 seconds after a blanking period of 3 months.Results The SNA originated from the right coronary artery in 34 patients (43.6%) and the left circumflex artery in 44 patients (56.4%).Freedom from AF and antiarrhythmic drugs (AADs) at 1 year was 67.9 % (53/78) for all patients.After 1 year follow-up,79.4% (27/34) in right SNA group and 59.1% (26/44) in left SNA group (P=0.042) were in sinus rhythm.On multivariate analysis,left atrium size (HR=1.451,95%CI:1.240-1.697,P <0.001) and a left SNA (HR=6.22,95%CI:2.01-19.25,P=0.002)were the independent predictors of AF recurrence.Conclusions The left SNA is more frequent in the patients with paroxysmal AF.After one year follow-up,the presence of a left SNA was identified as an independent predictor of AF recurrence after CPVA in

  13. Cryopreservation of Prunus avium L. embryogenic tissues.

    Science.gov (United States)

    Grenier-de March, Ghislaine; de Boucaud, Marie-Therese; Chmielarz, Pawel

    2005-01-01

    Embryogenic tissues from wild cherry (Prunus avium L.) were successfully cryopreserved by using a one-step freezing procedure. Cryoprotection consisted of a pretreatment on solid medium with increasing sucrose concentrations (0.25 M for 1 day, 0.5 M for 1 day, 0.75 M for 2 days, and 1.0 M for 3 days), followed by air desiccation to about 20 percent moisture content (fresh weight basis). This method was compared with a pretreatment on solid medium containing 5 percent DMSO and 2 percent proline, followed by immersion in a modified PVS2 cryoprotective solution. Pretreatment on solid medium with increasing concentrations of sucrose led to regrowth of frozen embryogenic tissues, and after 6 weeks of culture, growth was comparable to that of non-dehydrated and non-frozen tissues. By contrast, no regrowth was observed when embryogenic tissues were submitted to the solid/liquid pretreatment with DMSO/proline and a modified PVS2 solution.

  14. A single or multistage mycobacterium avium subsp. paratuberculosis subunit vaccine

    DEFF Research Database (Denmark)

    2014-01-01

    The present invention provides one or more immunogenic polypeptides for use in a preventive or therapeutic vaccine against latent or active infection in a human or animal caused by a Mycobacterium species, e.g. Mycobacterium avium subsp. paratuberculosis. Furthermore a single or multi-phase vaccine...... comprising the one or more immunogenic polypeptides is provided for administration for the prevention or treatment of infection with a Mycobacterium species, e.g. Mycobacterium avium subsp. paratuberculosis. Additionally, nucleic acid vaccines, capable of in vivo expression of the multi-phase vaccine...

  15. Sequence similarities of the capsid gene of Chilean and European isolates of infectious pancreatic necrosis virus point towards a common origin.

    Science.gov (United States)

    Mutoloki, Stephen; Evensen, Øystein

    2011-07-01

    The Chilean salmonid industry was developed by importing breeding materials, a practice still in effect due to deficits in the national supply of roe. Importation of breeding materials is often associated with the transmission of pathogens. The objectives of this study were to compare the infectious pancreatic necrosis virus (IPNV) isolates from Chile to those of European origin and to determine the diversity of the Chilean IPNV. The VP2 genes of IPNV from Chilean fish (whose eggs originated from Scotland, Iceland and Norway) were compared to isolates from fish in Norway and Ireland. The results show that the isolates are identical (97-99%) and cluster into one genogroup. Our findings support previous reports of association between the trade-in breeding materials and transmission of pathogens. Furthermore, our results demonstrate the genotypic diversity of Chilean IPNV isolates. These findings have important implications for IPNV disease diagnosis and control in Chile.

  16. Comparative genomics among Saccharomyces cerevisiae × Saccharomyces kudriavzevii natural hybrid strains isolated from wine and beer reveals different origins

    Directory of Open Access Journals (Sweden)

    Peris David

    2012-08-01

    Full Text Available Abstract Background Interspecific hybrids between S. cerevisiae × S. kudriavzevii have frequently been detected in wine and beer fermentations. Significant physiological differences among parental and hybrid strains under different stress conditions have been evidenced. In this study, we used comparative genome hybridization analysis to evaluate the genome composition of different S. cerevisiae × S. kudriavzevii natural hybrids isolated from wine and beer fermentations to infer their evolutionary origins and to figure out the potential role of common S. kudriavzevii gene fraction present in these hybrids. Results Comparative genomic hybridization (CGH and ploidy analyses carried out in this study confirmed the presence of individual and differential chromosomal composition patterns for most S. cerevisiae × S. kudriavzevii hybrids from beer and wine. All hybrids share a common set of depleted S. cerevisiae genes, which also are depleted or absent in the wine strains studied so far, and the presence a common set of S. kudriavzevii genes, which may be associated with their capability to grow at low temperatures. Finally, a maximum parsimony analysis of chromosomal rearrangement events, occurred in the hybrid genomes, indicated the presence of two main groups of wine hybrids and different divergent lineages of brewing strains. Conclusion Our data suggest that wine and beer S. cerevisiae × S. kudriavzevii hybrids have been originated by different rare-mating events involving a diploid wine S. cerevisiae and a haploid or diploid European S. kudriavzevii strains. Hybrids maintain several S. kudriavzevii genes involved in cold adaptation as well as those related to S. kudriavzevii mitochondrial functions.

  17. Tuberculosis in Birds: Insights into the Mycobacterium avium Infections

    Directory of Open Access Journals (Sweden)

    Kuldeep Dhama

    2011-01-01

    Full Text Available Tuberculosis, a List B disease of World Organization for Animal Health, caused by M. avium or M. genavense predominantly affects poultry and pet or captive birds. Clinical manifestations in birds include emaciation, depression and diarrhea along with marked atrophy of breast muscle. Unlike tuberculosis in animals and man, lesions in lungs are rare. Tubercular nodules can be seen in liver, spleen, intestine and bone marrow. Granulomatous lesion without calcification is a prominent feature. The disease is a rarity in organized poultry sector due to improved farm practices, but occurs in zoo aviaries. Molecular techniques like polymerase chain reaction combined with restriction fragment length polymorphism and gene probes aid in rapid identification and characterization of mycobacteria subspecies, and overcome disadvantages of conventional methods which are slow, labour intensive and may at times fail to produce precise results. M. avium subsp. avium with genotype IS901+ and IS1245+ causes infections in animals and human beings too. The bacterium causes sensitivity in cattle to the tuberculin test. The paper discusses in brief the M. avium infection in birds, its importance in a zoonotic perspective, and outlines conventional and novel strategies for its diagnosis, prevention and eradication in domestic/pet birds and humans alike.

  18. Investigación de Mycobacterium avium subsp. paratuberculosis en leche ultrapasteurizada para consumo humano Research of Mycobacterium avium subsp. paratuberculosis in ultrapasteurized milk for human consumption

    Directory of Open Access Journals (Sweden)

    G. G Magnano

    2009-12-01

    Full Text Available Actualmente se vincula al Mycobacterium avium subsp. paratuberculosis como potencial agente etiológico implicado en la enfermedad de Crohn en humanos. Una de las vías de ingreso sería a través de la ingestión de leche contaminada. El objetivo fue evaluar la presencia de Map en leche comercial homogeneizada y ultrapasteurizada para consumo humano en supermercados de la ciudad de Río Cuarto, Córdoba, Argentina. Se muestrearon 98 envases de 1 litro de leche entera homogeneizada y ultrapasteurizada de seis marcas comerciales. Previa descontaminación con el método de Cornell modificado, se sembraron en medio de cultivo Herrold con y sin micobactina. Todas las muestras fueron negativas. Como posibles causas de estos resultados se discuten: el origen de la leche y su probable muy baja carga de micobacterias, la eficacia de la pasteurización, el proceso en el laboratorio, entre otras.Currently, Mycobacterium avium subsp. paratuberculosis is linked to Crohn's disease in humans as a potential etiologic agent. One route of infection to be considered is by the ingestion of contaminated milk. The objective of the present work was to evaluate Map's presence in commercial homogenized and ultrapasteurized milk for human consumption in supermarkets in the city of Río Cuarto, Córdoba, Argentina. Ninety eight packages of 1 liter of entire, homogenized and ultrapasteurized milk of six commercial brands were sampled. After decontamination by the modified Cornell's method, the samples were cultured in Herrold's medium with and without micobactin. All samples were negative. Possible causes of this result such as the origin of the milk and its probable very low amount of micobacterias, the efficiency of the pasteurization, the processing in the laboratory, among others are here discussed.

  19. Genotypic diversity and virulence markers of Yersinia enterocolitica biotype 1A strains isolated from clinical and non-clinical origins.

    Science.gov (United States)

    Campioni, Fábio; Falcão, Juliana P

    2014-03-01

    Yersinia enterocolitica biotype 1A (B1A) strains are considered as non-pathogenic; however, some reports have identified some strains as the causal agents of infection. In South America, few studies molecularly characterized the strains of this biotype. This work typed 51 B1A strains isolated from clinical and non-clinical sources from Brazil and Chile by Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) to elucidate their genotypic diversity, and verify the distribution of 11 virulence markers by PCR. The strains were divided into two groups, ERIC-A and ERIC-B, clustered independently of their clinical or non-clinical origin. No differences were observed in the frequencies of the virulence markers between clinical and non-clinical strains. However, the genes ystB, hreP and myfA occurred exclusively in the strains of the group ERIC-A. Some clinical and non-clinical strains were clustered in the same genetic group and presented the same number of virulence markers, which might suggest the role of the environment and food as a potential source of infection for humans and animals. The results corroborate with the hypothesis that B1A strains are divided into two main clusters that differ in the frequency of some virulence markers, a fact observed for the first time in South American strains.

  20. Phylogenetic analysis of spring virema of carp virus reveals distinct subgroups with common origins for recent isolates in North America and the UK.

    Science.gov (United States)

    Miller, O; Fuller, F J; Gebreyes, W A; Lewbart, G A; Shchelkunov, I S; Shivappa, R B; Joiner, C; Woolford, G; Stone, D M; Dixon, P F; Raley, M E; Levine, J F

    2007-07-16

    Genetic relationships between 35 spring viremia of carp virus (SVCV) genogroup Ia isolates were determined based on the nucleotide sequences of the phosphoprotein (P) gene and glycoprotein (G) genes. Phylogenetic analysis based on P gene sequences revealed 2 distinct subgroups within SVCV genogroup Ia, designated SVCV Iai and Iaii, and suggests at least 2 independent introductions of the virus into the USA in 2002. Combined P- and G-sequence data support the emergence of SVCV in Illinois, USA, and in Lake Ontario, Canada, from the initial outbreak in Wisconsin, USA, and demonstrate a close genetic link to viruses isolated during routine import checks on fish brought into the UK from Asia. The data also showed a genetic link between SVCV isolations made in Missouri and Washington, USA, in 2004 and the earlier isolation made in North Carolina, USA, in 2002. However, based on the close relationship to a 2004 UK isolate, the data suggest than the Washington isolate represents a third introduction into the US from a common source, rather than a reemergence from the 2002 isolate. There was strong phylogenetic support for an Asian origin for 9 of 16 UK viruses isolated either from imported fish, or shown to have been in direct contact with fish imported from Asia. In one case, there was 100% nucleotide identity in the G-gene with a virus isolated in China.

  1. Detection of Mycobacterium avium subsp. paratuberculosis in Milk from Clinically Affected Cows by PCR and culture

    DEFF Research Database (Denmark)

    Giese, Steen Bjørck; Ahrens, Peter

    1999-01-01

    -negative on intestinal mucosa, but culture-positive in milk, and both faeces and milk were negative in culture and PCR from 2 cows. In conclusion the presence of M. a. paratuberculosis could be detected in raw milk by PCR but cultivation of milk was more sensitive in detecting the organism.......Milk and faecal samples from cows with clinical symptoms of paratuberculosis were examined for the presence of Mycobacterium avium subsp.paratuberculosis (M. a. paratuberculosis) by culture and PCR. M. a. paratuberculosis was isolated in varied numbers from faeces or intestinal mucosa in 8 of 11...... animals. In milk from 5 cows (all faecal culture-positive) we cultivated a few colonies of M. a. paratuberculosis (less than 100 CFU per mi). Milk samples from 2 cows were PCR-positive (both animals were faecal culture-positive, and 1 cow was milk culture positive). One cow was culture...

  2. Mycobacterium avium complex pulmonary disease: characteristics and treatment in an Irish patient cohort.

    LENUS (Irish Health Repository)

    Judge, EP

    2016-04-01

    The prevalence of Mycobacterium avium complex (MAC) pulmonary disease is increasing globally. However, reliable national and international data relating to its epidemiology and management is lacking. During the period 2003-2014, MAC was isolated from the pulmonary samples of 75 patients at the Irish Mycobacteria Reference Laboratory (IMRL). Most patients (42, 56%) had underlying pulmonary disease, and 37 (49%) had clinical\\/radiographic characteristics consistent with MAC pulmonary disease. However, only 18 patients (24%) fulfilled internationally accepted criteria for diagnosis\\/treatment of this disease. Treatment was started in 13 (72%) of these cases, which is similar to internationally published treatment rates. The diagnosis of significant MAC pulmonary disease can be difficult, and treatment is not always warranted even when diagnostic criteria are met.

  3. Variation in Fumonisin and Ochratoxin Production Associated with Differences in Biosynthetic Gene Content in Aspergillus niger and A. welwitschiae Isolates from Multiple Crop and Geographic Origins

    Science.gov (United States)

    Susca, Antonia; Proctor, Robert H.; Morelli, Massimiliano; Haidukowski, Miriam; Gallo, Antonia; Logrieco, Antonio F.; Moretti, Antonio

    2016-01-01

    The fungi Aspergillus niger and A. welwitschiae are morphologically indistinguishable species used for industrial fermentation and for food and beverage production. The fungi also occur widely on food crops. Concerns about their safety have arisen with the discovery that some isolates of both species produce fumonisin (FB) and ochratoxin A (OTA) mycotoxins. Here, we examined FB and OTA production as well as the presence of genes responsible for synthesis of the mycotoxins in a collection of 92 A. niger/A. welwitschiae isolates from multiple crop and geographic origins. The results indicate that (i) isolates of both species differed in ability to produce the mycotoxins; (ii) FB-nonproducing isolates of A. niger had an intact fumonisin biosynthetic gene (fum) cluster; (iii) FB-nonproducing isolates of A. welwitschiae exhibited multiple patterns of fum gene deletion; and (iv) OTA-nonproducing isolates of both species lacked the ochratoxin A biosynthetic gene (ota) cluster. Analysis of genome sequence data revealed a single pattern of ota gene deletion in the two species. Phylogenetic analysis suggest that the simplest explanation for this is that ota cluster deletion occurred in a common ancestor of A. niger and A. welwitschiae, and subsequently both the intact and deleted cluster were retained as alternate alleles during divergence of the ancestor into descendent species. Finally, comparison of results from this and previous studies indicate that a majority of A. niger isolates and a minority of A. welwitschiae isolates can produce FBs, whereas, a minority of isolates of both species produce OTA. The comparison also suggested that the relative abundance of each species and frequency of FB/OTA-producing isolates can vary with crop and/or geographic origin. PMID:27667988

  4. Modeling of Mycobacterium avium subsp. paratuberculosis in farm bulk tank milk

    DEFF Research Database (Denmark)

    Okura, Hisako; Nielsen, Søren Saxmose; Toft, Nils

    2012-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) in milk of bovine origin is suspected of being implicated Crohn’s disease in humans. Pasteurization is considered to reduce the concentration of MAP by at least 4 to 5 log10. This study aimed at estimating the level of MAP in milk at farm level...... super-shedders. At the prevalence of 15%, discarding milk from test positive cows would result in discarding 11% of milk and reduce the MAP level by 80%. The model was relatively simple yet capable of capturing true infection status and associated contributions from milk and feces. Further knowledge...

  5. Complete Genome Sequence of Micromonospora Strain L5, a Potential Plant-Growth-Regulating Actinomycete, Originally Isolated from Casuarina equisetifolia Root Nodules

    Energy Technology Data Exchange (ETDEWEB)

    Hirsch, A. M.; Alvarado, J.; Bruce, D.; Chertkov, O.; De Hoff, P. L.; Detter, J. C.; Fujishige, N. A.; Goodwin, L. A.; Han, J.; Han, S.; Ivanova, N.; Land, M. L.; Lum, M. R.; Milani-Nejad, N.; Nolan, M.; Pati, A.; Pitluck, S.; Tran, S. S.; Woyke, T.; Valdes, M.

    2013-08-29

    Micromonospora species live in diverse environments and exhibit a broad range of functions including antibiotic production, biocontrol, and ability to degrade complex polysaccharides. To learn more about these versatile actinomycetes, we sequenced the genome of strain L5, originally isolated from root nodules of an actinorhizal plant growing in Mexico.

  6. Fate of Mycobacterium avium subsp. paratuberculosis in Swiss Hard and Semihard Cheese Manufactured from Raw Milk

    OpenAIRE

    Spahr, U.; Schafroth, K.

    2001-01-01

    Raw milk was artificially contaminated with declumped cells of Mycobacterium avium subsp. paratuberculosis at a concentration of 104 to 105 CFU/ml and was used to manufacture model hard (Swiss Emmentaler) and semihard (Swiss Tisliter) cheese. Two different strains of M. avium subsp. paratuberculosis were tested, and for each strain, two model hard and semihard cheeses were produced. The survival of M. avium subsp. paratuberculosis cells was monitored over a ripening period of 120 days by plat...

  7. MLVA-16 typing of 295 marine mammal Brucella isolates from different animal and geographic origins identifies 7 major groups within Brucella ceti and Brucella pinnipedialis

    Directory of Open Access Journals (Sweden)

    Jacques Isabelle

    2009-07-01

    Full Text Available Abstract Background Since 1994, Brucella strains have been isolated from a wide range of marine mammals. They are currently recognized as two new Brucella species, B. pinnipedialis for the pinniped isolates and B. ceti for the cetacean isolates in agreement with host preference and specific phenotypic and molecular markers. In order to investigate the genetic relationships within the marine mammal Brucella isolates and with reference to terrestrial mammal Brucella isolates, we applied in this study the Multiple Loci VNTR (Variable Number of Tandem Repeats Analysis (MLVA approach. A previously published assay comprising 16 loci (MLVA-16 that has been shown to be highly relevant and efficient for typing and clustering Brucella strains from animal and human origin was used. Results 294 marine mammal Brucella strains collected in European waters from 173 animals and a human isolate from New Zealand presumably from marine origin were investigated by MLVA-16. Marine mammal Brucella isolates were shown to be different from the recognized terrestrial mammal Brucella species and biovars and corresponded to 3 major related groups, one specific of the B. ceti strains, one of the B. pinnipedialis strains and the last composed of the human isolate. In the B. ceti group, 3 subclusters were identified, distinguishing a cluster of dolphin, minke whale and porpoise isolates and two clusters mostly composed of dolphin isolates. These results were in accordance with published analyses using other phenotypic or molecular approaches, or different panels of VNTR loci. The B. pinnipedialis group could be similarly subdivided in 3 subclusters, one composed exclusively of isolates from hooded seals (Cystophora cristata and the two others comprising other seal species isolates. Conclusion The clustering analysis of a large collection of marine mammal Brucella isolates from European waters significantly strengthens the current view of the population structure of these two

  8. Short communication: isolation and phylogenetic analysis of an avian-origin H3N2 canine influenza virus in dog shelter, China.

    Science.gov (United States)

    Su, Shuo; Yuan, Ziguo; Chen, Jidang; Xie, Jiexiong; Li, Huatao; Huang, Zhen; Zhang, Minze; Du, Guohao; Chen, Zhongming; Tu, Liqing; Zou, Yufei; Miao, Junhao; Wang, Hui; Jia, Kun; Li, Shoujun

    2013-06-01

    A H3N2 canine influenza virus, A/canine/Guangdong/3/2011 (H3N2), was isolated from roaming dogs in rural China. Sequence and phylogenetic analysis of eight gene segments revealed that the A/canine/Guangdong/3/2011 (H3N2) was most similar to a recent H3N2 canine influenza virus isolated in cats from South Korea, which originated from an avian strain. To our knowledge, this is the first report of an avian-origin H3N2 CIV which was isolated from roaming dogs in China. The epidemiologic information provided herein suggests that continued study is required to determine if this virus could be established in the roaming dog population in rural China and pose potential threats to public health.

  9. Characterization of Listeria monocytogenes isolates of food and human origins from Brazil using molecular typing procedures and in vitro cell culture assays.

    Science.gov (United States)

    Bueno, Valter F; Banerjee, Pratik; Banada, Padmapriya P; José de Mesquita, Albenones; Lemes-Marques, Eneida G; Bhunia, Arun K

    2010-02-01

    The spreading of diseases through foods is a worldwide concern. Here, molecular and in vitro cell-culture assays were employed to characterize 63 Brazilian Listeria monocytogenes isolates (food, 47; clinical, 16). Serotype 4b was the most predominant (49%) followed by (1/2)b (30%), (1/2)a (10%), (1/2)c (6%), 3c (3%) and 3b (2%). Ribotyping yielded 17 ribopatterns, which were grouped into four phylogenetic clusters. Cluster A comprised of 39/63 isolates primarily of food origin, and clusters B, C and D contained both food and clinical isolates. Isolates were positive for virulence determinants prfA, hlyA and inlA: clinical isolates were more invasive to Caco-2 cells and expressed high levels of inlA transcripts than the food isolates. Highly invasive isolates also provoked more Ped-2E9 cells to die by apoptosis than the weakly-invasive strains. These data demonstrate a strong genetic relatedness among clinical and food isolates and suggest transmission of a subset of L. monocytogenes strains from food to humans.

  10. Transcriptional analysis of diverse strains Mycobacterium avium subspecies paratuberculosis in primary bovine monocyte derived macrophages.

    Science.gov (United States)

    Zhu, Xiaochun; Tu, Zheng J; Coussens, Paul M; Kapur, Vivek; Janagama, Harish; Naser, Saleh; Sreevatsan, Srinand

    2008-10-01

    In this study we analyzed the macrophage-induced gene expression of three diverse genotypes of Mycobacterium avium subsp. paratuberculosis (MAP). Using selective capture of transcribed sequences (SCOTS) on three genotypically diverse MAP isolates from cattle, human, and sheep exposed to primary bovine monocyte derived macrophages for 48 h and 120 h we created and sequenced six cDNA libraries. Sequence annotations revealed that the cattle isolate up-regulated 27 and 241 genes; the human isolate up-regulated 22 and 53 genes, and the sheep isolate up-regulated 35 and 358 genes, at the two time points respectively. Thirteen to thirty-three percent of the genes identified did not have any annotated function. Despite variations in the genes identified, the patterns of expression fell into overlapping cellular functions as inferred by pathway analysis. For example, 10-12% of the genes expressed by all three strains at each time point were associated with cell-wall biosynthesis. All three strains of MAP studied up-regulated genes in pathways that combat oxidative stress, metabolic and nutritional starvation, and cell survival. Taken together, this comparative transcriptional analysis suggests that diverse MAP genotypes respond with similar modus operandi for survival in the host.

  11. Ecology and genomic features of infection with Mycobacterium avium subspecies paratuberculosis in Egypt.

    Science.gov (United States)

    Amin, Adel S; Hsu, Chung-Yi; Darwish, Samah F; Ghosh, Pallab; AbdEl-Fatah, Eman M; Behour, Tahani S; Talaat, Adel M

    2015-04-01

    Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is the causative agent of paratuberculosis, or Johne's disease, in cattle, with potential involvement in cases of Crohn's disease in humans. Johne's disease is found worldwide and is economically important for both beef and dairy industries. In an effort to characterize this important infection in Egypt, we analysed the ecological and genomic features of recent isolates of M. paratuberculosis. In this report, we examined 26 Holstein dairy herds distributed throughout Egypt, from 2010 to 2013. Using PCR analysis of faecal samples, we estimated a mean herd-level prevalence of 65.4 %, with animal-level infection that reached a mean of 13.6 % among animals suffering from diarrhoea. Whole genome sequencing of field isolates identified numerous single nucleotide polymorphisms among field isolates relative to the standard M. paratuberculosis K10 genome. Interestingly, the virulence of M. paratuberculosis isolates from Egypt revealed diverse virulence phenotypes in the murine model of paratuberculosis, with significant differences in tissue colonization, particularly during the chronic stage of infection. Overall, our analysis confirmed that Johne's disease is a newly identified problem in Egypt and indicated that M. paratuberculosis has potentially diverse genotypes that impact its virulence. Further ecological mapping and genomic analysis of M. paratuberculosis will enhance our understanding of the transmission and evolutionary dynamics of this pathogen under natural field conditions.

  12. Mycobacterium avium complex organisms predominantly colonize in the bathtub inlets of patients' bathrooms.

    Science.gov (United States)

    Nishiuchi, Yukiko; Tamura, Aki; Kitada, Seigo; Taguri, Takahiro; Matsumoto, Sohkichi; Tateishi, Yoshitaka; Yoshimura, Mamiko; Ozeki, Yuriko; Matsumura, Narumi; Ogura, Hisashi; Maekura, Ryoji

    2009-05-01

    Medical treatment of pulmonary Mycobacterium avium complex (MAC) disease does not always provide curative effects and is frequently hampered by recurrence. This suggests the presence of a reservoir for MAC in the environment surrounding patients. We previously reported the recovery of MAC isolates from the residential bathrooms of outpatients. In the present study, to ascertain the colonizing sites and the possibility of an MAC reservoir in the bathrooms of patients, we tested the recovery and the genetic diversity of MAC isolates from 6 sites of specimens, including 2 additional sampling sites, inside the showerhead and the bathtub inlet, in the residential bathrooms of patients with pulmonary MAC disease. MAC isolates were recovered from 15 out of the 29 bathrooms (52%), including specimens from 14 bathtub inlets and 3 showerheads. Nearly half of these bathrooms (7/15) contained MAC strains that were identical or similar to their respective clinical isolates Additionally, in 5 out of 15 bathrooms, polyclonal colonization was revealed by pulsed-field gel electrophoresis. The results imply that colonization of MAC organisms in the bathrooms of MAC patients occurs predominantly in the bathtub inlets, and there is thus a risk of infection and/or reinfection for patients via use of the bathtub and other sites in the bathroom.

  13. Isolation and identification of lactid acid bacteria originated from king grass (Pennisetum purpureophoides) as candidate of probiotic for livestock

    OpenAIRE

    Santoso B; Maunatin A; Hariadi BT; Abubakar H

    2013-01-01

    A study was conducted to isolate and identify strain of lactic acid bacteria (LAB) isolated from king grass, and to determine their potential as candidate of probiotic for livestock. The LAB was isolated by culturing king grass extract in De Man, Rogosa and Sharpe (MRS) medium. The pure culture LAB was used to identify strain of bacteria using Analytical Profile Index (API) 50 CH kit. The result showed that the strain bacteria was identified as Lactobacillus plantarum. L. plantarum was able t...

  14. Short communication: Subtyping of Staphylococcus haemolyticus isolates from milk and corresponding teat apices to verify the potential teat-skin origin of intramammary infections in dairy cows.

    Science.gov (United States)

    Leroy, Frédéric; Van Coillie, Els; Braem, Gorik; Piessens, Veerle; Verbist, Bert; De Vuyst, Luc; De Vliegher, Sarne

    2015-11-01

    Coagulase-negative staphylococci (CNS) are a major cause of intramammary infections (IMI) in dairy cows and they colonize the teat skin. Staphylococcus haemolyticus, one of the more common CNS, has been identified as a highly versatile opportunistic species. The aim of the present study was to gain better insight into the adaptation of S. haemolyticus subtypes to the udder ecosystem with respect to IMI development. During a longitudinal observational study conducted over 13 mo on 6 Flemish dairy herds, S. haemolyticus isolates were recovered from milk and teat apices. A total of 44 S. haemolyticus isolates originating from milk (24 isolates) and teat apices (20 isolates) of 6 selected udder quarters were singled out and analyzed using a combined methodology of (GTG)5-PCR and amplified fragment length polymorphism (AFLP) fingerprinting to determine intraspecies differences. Combining both fingerprinting methods, 4 S. haemolyticus subtypes were obtained (I to IV). Subtypes I, II, and IV were recovered from both milk and teat apex samples and were found to be associated with persisting IMI. Subtype III, not apparently related to IMI, was isolated solely from teat apices and not from milk. In general, S. haemolyticus subtypes found in milk from infected quarters could be recovered from the corresponding teat apices, although the latter could be colonized with up to 3 different subtypes. Comparing subtypes from milk and teat apices indicates that the IMI-causing agent likely originates from the teat skin.

  15. Isolation and Genomic Characterization of a Duck-Origin GPV-Related Parvovirus from Cherry Valley Ducklings in China.

    Science.gov (United States)

    Chen, Hao; Dou, Yanguo; Tang, Yi; Zhang, Zhenjie; Zheng, Xiaoqiang; Niu, Xiaoyu; Yang, Jing; Yu, Xianglong; Diao, Youxiang

    2015-01-01

    A newly emerged duck parvovirus, which causes beak atrophy and dwarfism syndrome (BADS) in Cherry Valley ducks, has appeared in Northern China since March 2015. To explore the genetic diversity among waterfowl parvovirus isolates, the complete genome of an identified isolate designated SDLC01 was sequenced and analyzed in the present study. Genomic sequence analysis showed that SDLC01 shared 90.8%-94.6% of nucleotide identity with goose parvovirus (GPV) isolates and 78.6%-81.6% of nucleotide identity with classical Muscovy duck parvovirus (MDPV) isolates. Phylogenetic analysis of 443 nucleotides (nt) of the fragment A showed that SDLC01 was highly similar to a mule duck isolate (strain D146/02) and close to European GPV isolates but separate from Asian GPV isolates. Analysis of the left inverted terminal repeat regions revealed that SDLC01 had two major segments deleted between positions 160-176 and 306-322 nt compared with field GPV and MDPV isolates. Phylogenetic analysis of Rep and VP1 encoded by two major open reading frames of parvoviruses revealed that SDLC01 was distinct from all GPV and MDPV isolates. The viral pathogenicity and genome characterization of SDLC01 suggest that the novel GPV (N-GPV) is the causative agent of BADS and belongs to a distinct GPV-related subgroup. Furthermore, N-GPV sequences were detected in diseased ducks by polymerase chain reaction and viral proliferation was demonstrated in duck embryos and duck embryo fibroblast cells.

  16. Pathotypic and genotypic characterization of two Bangladeshi isolates of Newcastle disease virus of chicken and pigeon origin.

    Science.gov (United States)

    Nooruzzaman, M; Mazumder, A C; Khatun, S; Chowdhury, E H; Das, P M; Islam, M R

    2015-02-01

    Two Bangladeshi isolates of Newcastle disease virus (NDV), one from a chicken and one from a pigeon, were characterized in this study. Pathogenicity of the isolates was evaluated on the basis of intracerebral pathogenicity index (ICPI). Both the isolates were found to be of velogenic pathotype having ICPI of 1.83 and 1.51 for the chicken and pigeon isolate, respectively. Genotype of the isolates was determined by phylogenetic analysis based on partial F gene sequences. A 766-bp genome fragment spanning partial M and F gene was amplified by RT-PCR and sequenced. The first 354 bp of the coding region of F gene and corresponding deduced amino acid sequences (residues 1-118) of these two NDV isolates were aligned with that of other NDV strains retrieved from GenBank. A phylogenetic tree constructed from the alignment showed that the chicken isolate (BD-C162) belonged to the newly described genotype XIII and the pigeon isolate (BD-P01) to genotype VI. Both the chicken and pigeon isolates possessed a virulent-like fusion protein cleavage site (112) RRQKRF(117) .

  17. Mycobacterium avium Possesses Extracellular DNA that Contributes to Biofilm Formation, Structural Integrity, and Tolerance to Antibiotics.

    Science.gov (United States)

    Rose, Sasha J; Babrak, Lmar M; Bermudez, Luiz E

    2015-01-01

    Mycobacterium avium subsp. hominissuis is an opportunistic pathogen that is associated with biofilm-related infections of the respiratory tract and is difficult to treat. In recent years, extracellular DNA (eDNA) has been found to be a major component of bacterial biofilms, including many pathogens involved in biofilm-associated infections. To date, eDNA has not been described as a component of mycobacterial biofilms. In this study, we identified and characterized eDNA in a high biofilm-producing strain of Mycobacterium avium subsp. hominissuis (MAH). In addition, we surveyed for presence of eDNA in various MAH strains and other nontuberculous mycobacteria. Biofilms of MAH A5 (high biofilm-producing strain) and MAH 104 (reference strain) were established at 22°C and 37°C on abiotic surfaces. Acellular biofilm matrix and supernatant from MAH A5 7 day-old biofilms both possess abundant eDNA, however very little eDNA was found in MAH 104 biofilms. A survey of MAH clinical isolates and other clinically relevant nontuberculous mycobacterial species revealed many species and strains that also produce eDNA. RAPD analysis demonstrated that eDNA resembles genomic DNA. Treatment with DNase I reduced the biomass of MAH A5 biofilms when added upon biofilm formation or to an already established biofilm both on abiotic surfaces and on top of human pharyngeal epithelial cells. Furthermore, co-treatment of an established biofilm with DNase 1 and either moxifloxacin or clarithromycin significantly increased the susceptibility of the bacteria within the biofilm to these clinically used antimicrobials. Collectively, our results describe an additional matrix component of mycobacterial biofilms and a potential new target to help treat biofilm-associated nontuberculous mycobacterial infections.

  18. Sensitive and specific enzyme-linked immunosorbent assay for detecting serum antibodies against Mycobacterium avium subsp. paratuberculosis in fallow deer.

    Science.gov (United States)

    Prieto, José M; Balseiro, Ana; Casais, Rosa; Abendaño, Naiara; Fitzgerald, Liam E; Garrido, Joseba M; Juste, Ramon A; Alonso-Hearn, Marta

    2014-08-01

    The enzyme-linked immunosorbent assay (ELISA) is the diagnostic test most commonly used in efforts to control paratuberculosis in domestic ruminants. However, commercial ELISAs have not been validated for detecting antibodies against Mycobacterium avium subsp. paratuberculosis in wild animals. In this study, we compared the sensitivities and specificities of five ELISAs using individual serum samples collected from 41 fallow deer with or without histopathological lesions consistent with paratuberculosis. Two target antigenic preparations were selected, an ethanol-treated protoplasmic preparation obtained from a fallow deer M. avium subsp. paratuberculosis isolate (ELISAs A and B) and a paratuberculosis protoplasmic antigen (PPA3) (ELISAs C and D). Fallow deer antibodies bound to the immobilized antigens were detected by using a horseradish peroxidase (HRP)-conjugated anti-fallow deer IgG antibody (ELISAs A and C) or HRP-conjugated protein G (ELISAs B and D). A commercially available assay, ELISA-E, which was designed to detect M. avium subsp. paratuberculosis antibodies in cattle, sheep, and goats, was also tested. Although ELISAs A, C, and E had the same sensitivity (72%), ELISAs A and C were more specific (100%) for detecting fallow deer with lesions consistent with paratuberculosis at necropsy than was the ELISA-E (87.5%). In addition, the ELISA-A was particularly sensitive for detecting fallow deer in the latent stages of infection (62.5%). The antibody responses detected with the ELISA-A correlated with both the severity of enteric lesions and the presence of acid-fast bacteria in gut tissue samples. In summary, our study shows that the ELISA-A can be a cost-effective diagnostic tool for preventing the spread of paratuberculosis among fallow deer populations.

  19. Variable agreement among experts regarding Mycobacterium avium complex lung disease.

    Science.gov (United States)

    Marras, Theodore K; Prevots, D Rebecca; Jamieson, Frances B; Winthrop, Kevin L

    2015-02-01

    Data regarding many clinical aspects of pulmonary Mycobacterium avium complex (pMAC) are lacking. Guidelines rely substantially upon expert opinion, integrated through face-to-face meetings, variably weighting individual opinions. We surveyed North American non-tuberculous mycobacteria experts regarding clinical aspects of pMAC using Delphi methods. Nineteen of 26 invited experts (73%) responded, with extensive variability. Convergence could not be reached for most questions. Respondents described extensive uncertainty around specific issues. Findings underscore urgent need for more research.

  20. 欧洲甜樱桃研究进展及开发利用前景%Research Progress of Prunus avium and the Development Prospects

    Institute of Scientific and Technical Information of China (English)

    兰士波

    2012-01-01

    在汲取国内外相关研究成果及学术观点的基础上,详细阐述了欧洲甜樱桃的生物学和生态学特性、起源及地理分布,探讨了果实所含营养成分及其营养和医疗保健价值;分析了存量资源的保存现状、优良品种及砧木选育的研究进展;指出了资源开发利用过程中存在的不足,展望了欧洲甜樱桃系列产品综合开发和创新利用的广阔前景。旨在实现欧洲甜樱桃无公害、标准化栽培及资源的可持续利用,为推动功能性饮品、保健品及药品的产业化发展,提供理论依据和科技支撑。%This paper expounded systematically the biological and ecological characteristics of Prunus avium L.,and its the origin and geographic distribution on the basis of generalized the research production and science paper of P.avium at home and abroad in the recently years.We discussed the nutrition ingredient of P.avium and its nutrition and health function,analyzed the status quo of germplasm resources conservation,utilization and estimate,and the progress of variety breeding study.We pointed out the insufficiencies in exploitation and utilization,and expected the bright future for the development of series production of P.avium The aim was to carry out nuisanceless,standardization planting and sustainable utilization of P.avium resources,provided theory warranty and support of science and technology for promoted the industrialization development of functionality beverage,health products and material medica of P.avium.

  1. Coagulase-negative staphylococci (CoNS) isolated from ready-to-eat food of animal origin--phenotypic and genotypic antibiotic resistance.

    Science.gov (United States)

    Chajęcka-Wierzchowska, Wioleta; Zadernowska, Anna; Nalepa, Beata; Sierpińska, Magda; Łaniewska-Trokenheim, Łucja

    2015-04-01

    The aim of this work was to study the pheno- and genotypical antimicrobial resistance profile of coagulase negative staphylococci (CoNS) isolated from 146 ready-to-eat food of animal origin (cheeses, cured meats, sausages, smoked fishes). 58 strains were isolated, they were classified as Staphylococcus xylosus (n = 29), Staphylococcus epidermidis (n = 16); Staphylococcus lentus (n = 7); Staphylococcus saprophyticus (n = 4); Staphylococcus hyicus (n = 1) and Staphylococcus simulans (n = 1) by phenotypic and genotypic methods. Isolates were tested for resistance to erythromycin, clindamycin, gentamicin, cefoxitin, norfloxacin, ciprofloxacin, tetracycline, tigecycline, rifampicin, nitrofurantoin, linezolid, trimetoprim, sulphamethoxazole/trimethoprim, chloramphenicol, quinupristin/dalfopristin by the disk diffusion method. PCR was used for the detection of antibiotic resistance genes encoding: methicillin resistance--mecA; macrolide resistance--erm(A), erm(B), erm(C), mrs(A/B); efflux proteins tet(K) and tet(L) and ribosomal protection proteins tet(M). For all the tet(M)-positive isolates the presence of conjugative transposons of the Tn916-Tn1545 family was determined. Most of the isolates were resistant to cefoxitin (41.3%) followed by clindamycin (36.2%), tigecycline (24.1%), rifampicin (17.2%) and erythromycin (13.8%). 32.2% staphylococcal isolates were multidrug resistant (MDR). All methicillin resistant staphylococci harboured mecA gene. Isolates, phenotypic resistant to tetracycline, harboured at least one tetracycline resistance determinant on which tet(M) was most frequent. All of the isolates positive for tet(M) genes were positive for the Tn916-Tn1545 -like integrase family gene. In the erythromycin-resistant isolates, the macrolide resistance genes erm(C) or msr(A/B) were present. Although coagulase-negative staphylococci are not classical food poisoning bacteria, its presence in food could be of public health significance due to the possible spread of

  2. Effect of Prunus avium roots on river bank strong.

    Science.gov (United States)

    Bibalani, Ghassem Habibi; Bazhrang, Zia; Mohsenifar, Hani; Shibaei, Naeime; Joodi, Lila

    2008-04-15

    A pulling effect by side roots is one way in which roots help to side in-plane strong of a little depth soil mass. In contrast to the effect of vertically-enlarge roots, whereby soil is strengthened by an increase in its shear strength, the pulling effect strengthens the soil by increasing the tensile strength of the rooted soil zone. To verify whether or not a pulling effect exists in the root system of Prunus avium in the Roudsar, North Iran and to study the importance and size of this effect, a direct in situ test was led at a site in the Chaboksar Forests. The results from the site showed that, in the surface soil (0-30 cm), Side roots can provide a pull force of up to 490-712 N (Newtons) over a vertical cross-section area of 20-50 cm2, or an enhance in the pulling stability of the rooted soil by about 48.1%. The test results suggest that, together with the Prunus avium vertical roots, which keep the little depth rooted soil zone to the deep and more stable soil mass, the side roots of the Prunus avium, with their pulling effect, are able to make less against little depth instability in the forest slopes, such as little depth slide, to a certain degree.

  3. Antigenic Profiles of Recombinant Proteins from Mycobacterium avium subsp paratuberculosis in Sheep with Johne's Disease

    Science.gov (United States)

    Methods to improve the ELISA test to detect Mycobacterium avium subsp paratuberculosis have been explored over several years. Previously, selected recombinant proteins of M. avium subspecies paratuberculosis were found to be immunogenic in cattle with Johne’s disease. In the present study, antibo...

  4. Roles for Cell Wall Glycopeptidolipid in Surface Adherence and Planktonic Dispersal of Mycobacterium avium

    Science.gov (United States)

    The opportunistic pathogen Mycobacterium avium is a significant inhabitant of biofilms in drinking water distribution systems. M. avium expresses on its cell surface serovar-specific glycopeptidolipids (ssGPLs). Studies have implicated the core GPL in biofilm formation by M. aviu...

  5. Relationships between antimicrobial resistance, distribution of virulence factor genes and the origin of Trueperella pyogenes isolated from domestic animals and European bison (Bison bonasus).

    Science.gov (United States)

    Rzewuska, Magdalena; Czopowicz, Michał; Gawryś, Marta; Markowska-Daniel, Iwona; Bielecki, Wojciech

    2016-07-01

    Trueperella pyogenes is an opportunistic pathogen causing suppurative infections in livestock and wild animals. Although this bacterium is known for a long time, our knowledge about its pathogenicity is still insufficient. In this study the relationships between antimicrobial resistance profiles, distribution of virulence factor genes and the origin of T. pyogenes isolates were investigated. Isolates (n = 97) from various infections in domestic animals and European bison were studied. Minimal inhibitory concentrations of 12 antimicrobials were determined by a strip diffusion method, and PCR was used for detection of genes encoding seven putative virulence factors. All strains were susceptible to tested beta-lactams, and a statistically significant correlation between the resistance to enrofloxacin, tetracycline, macrolides, clindamycin, and a strain origin was found. The isolates from European bison were more susceptible than those from livestock, however the resistance to tetracycline and fluoroquinolones was observed. The plo and fimA genes were detected in all strains. There was no statistically significant association between the distribution of particular virulence factor genes and the type of infection, but the nanH, nanP and fimG genes were less frequently found in the isolates from European bison. The presence of three genes, nanP, nanH and cbpA, was found to be related to the resistance to tetracycline and ciprofloxacin. In conclusion, the resistance patterns of T. pyogenes were correlated with an isolate origin, but our findings did not allow to indicate which of the putative virulence factors may play a crucial role in the pathogenesis of particular types of T. pyogenes infection.

  6. Mycobacterium avium-intracellulare cellulitis occurring with septic arthritis after joint injection: a case report

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    Murdoch David M

    2007-02-01

    Full Text Available Abstract Background Cellulitis caused by Mycobacterium avium-intracellulare has rarely been described. Mycobacterium avium-intracellulare is a rare cause of septic arthritis after intra-articular injection, though the causative role of injection is difficult to ascertain in such cases. Case presentation A 57-year-old with rheumatoid arthritis treated with prednisone and azathioprine developed bilateral painful degenerative shoulder arthritis. After corticosteroid injections into both acromioclavicular joints, he developed bilateral cellulitis centered over the injection sites. Skin biopsy showed non-caseating granulomas, and culture grew Mycobacterium avium-intracellulare. Joint aspiration also revealed Mycobacterium avium-intracellulare infection. Conclusion Although rare, skin and joint infections caused by Mycobacterium avium-intracellulare should be considered in any immunocompromised host, particularly after intra-articular injection. Stains for acid-fast bacilli may be negative in pathologic samples even in the presence of infection; cultures of tissue specimens should always be obtained.

  7. Isolation and identification of lactid acid bacteria originated from king grass (Pennisetum purpureophoides as candidate of probiotic for livestock

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    Santoso B

    2013-06-01

    Full Text Available A study was conducted to isolate and identify strain of lactic acid bacteria (LAB isolated from king grass, and to determine their potential as candidate of probiotic for livestock. The LAB was isolated by culturing king grass extract in De Man, Rogosa and Sharpe (MRS medium. The pure culture LAB was used to identify strain of bacteria using Analytical Profile Index (API 50 CH kit. The result showed that the strain bacteria was identified as Lactobacillus plantarum. L. plantarum was able to survive in extreme condition at pH 2 and 0.3% bile salt. L. plantarum also survived against pathogenic bacteria i.e. Staphylococcus aureus, Escherechia coli and Salmonella thypi. It is concluded that L. plantarum isolated from king grass could potentially to be used as probiotic for livestock.

  8. Differential pathogenicity of five Streptococcus agalactiae isolates of diverse geographic origin in Nile tilapia (Oreochromis niloticus L.)

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    Streptococcus agalactiae is an emerging pathogen of fish and has caused significant morbidity amd mortality worldwide. The work in this study assessed whether pathogenic differences exist among isolates from different geographic locations. Nile tilapia (Oreochromis niloticus L.) were administered an...

  9. Endophytic bacteria in plant tissue culture: differences between easy- and difficult-to-propagate Prunus avium genotypes.

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    Quambusch, Mona; Pirttilä, Anna Maria; Tejesvi, Mysore V; Winkelmann, Traud; Bartsch, Melanie

    2014-05-01

    The endophytic bacterial communities of six Prunus avium L. genotypes differing in their growth patterns during in vitro propagation were identified by culture-dependent and culture-independent methods. Five morphologically distinct isolates from tissue culture material were identified by 16S rDNA sequence analysis. To detect and analyze the uncultivable fraction of endophytic bacteria, a clone library was established from the amplified 16S rDNA of total plant extract. Bacterial diversity within the clone libraries was analyzed by amplified ribosomal rDNA restriction analysis and by sequencing a clone for each identified operational taxonomic unit. The most abundant bacterial group was Mycobacterium sp., which was identified in the clone libraries of all analyzed Prunus genotypes. Other dominant bacterial genera identified in the easy-to-propagate genotypes were Rhodopseudomonas sp. and Microbacterium sp. Thus, the community structures in the easy- and difficult-to-propagate cherry genotypes differed significantly. The bacterial genera, which were previously reported to have plant growth-promoting effects, were detected only in genotypes with high propagation success, indicating a possible positive impact of these bacteria on in vitro propagation of P. avium, which was proven in an inoculation experiment.

  10. Cytokine responses of bovine macrophages to diverse clinical Mycobacterium avium subspecies paratuberculosis strains

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    Coussens Paul

    2006-02-01

    Full Text Available Abstract Background Mycobacterium avium subsp. paratuberculosis (MAP, the causative agent of Johne's disease (JD persistently infects and survives within the host macrophages. While it is established that substantial genotypic variation exists among MAP, evidence for the correlates that associate specific MAP genotypes with clinical or sub-clinical disease phenotypes is presently unknown. Thus we studied strain differences in intracellular MAP survival and host responses in a bovine monocyte derived macrophage (MDM system. Results Intracellular survival studies showed that a bovine MAP isolate (B1018 and a human MAP isolate (Hu6 persisted in relatively higher numbers when compared with a sheep MAP isolate (S7565 at 24-hr, 48-hr and 96-hr post infection (PI. MDMs stimulated with B1018 up-regulated IL-10 at the transcript level and down-regulated TNFα at the protein and transcript levels compared with stimulations by the S7565 and Hu6. MDMs infected with Hu6 showed a down regulatory pattern of IL-10 and TNFα compared to stimulations by S7565. Cells stimulated with B1018 and Hu6 had low levels of matrix metalloprotease-3 (MMP3 and high levels of tissue inhibitor of metalloprotease-1 (TIMP1 at 96-hr PI relative to MDMs stimulated by S7565. Conclusion Taken together, results suggest that the bovine (B1018 and the human (Hu6 MAP isolates lead to anti-inflammatory and anti-invasive pathways in the macrophage environment whereas the sheep (S7565 MAP isolate induces a pro-inflammatory pathway. Thus the infecting strain genotype may play a role in polarizing the host immune responses and dictate the clinicopathological outcomes in this economically important disease.

  11. Diagnosis and Molecular Characterization of Mycobacterium avium subsp. paratuberculosis from Dairy Cows in Colombia.

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    Fernández-Silva, J A; Abdulmawjood, A; Bülte, M

    2011-01-01

    The objective of this study was the serological, bacteriological and molecular diagnosis, as well as the molecular characterization of Mycobacterium avium subsp. paratuberculosis (Map) in adult cows of five Colombian dairy herds. Serum samples were tested by an indirect absorbed enzyme-linked immunosorbent assay (ELISA-C). All fecal samples were tested by pooled culture. After that, fecal samples of Map positive pools were tested individually by culture and polymerase chain reaction (PCR). In one herd, slurry and tissue samples from one animal were also taken and tested by PCR and culture. Map isolates were analyzed by the Multilocus Short Sequence Repeat (MLSSR) and the Mycobacterial Interspersed Repetitive Units-Variable Number of Tandem Repeats (MIRU-VNTR) methods. ELISA produced positive results in 1.8% (6/329) of the animals and 40% (2/5) of the herds. Four fecal, two tissue, and two slurry samples from a herd were Map positive by culture and PCR. MLSSR and MIRU-VNTR revealed two different strain profiles among eight Map isolates recovered. This study reports the first molecular characterization of Map in one dairy herd in Colombia, the limitations for individual diagnosis of subclinical Map infections in cattle, and the usefulness of pooled fecal samples and environmental sampling for Map diagnosis.

  12. Diagnosis and Molecular Characterization of Mycobacterium avium subsp. paratuberculosis from Dairy Cows in Colombia

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    J. A. Fernández-Silva

    2011-01-01

    Full Text Available The objective of this study was the serological, bacteriological and molecular diagnosis, as well as the molecular characterization of Mycobacterium avium subsp. paratuberculosis (Map in adult cows of five Colombian dairy herds. Serum samples were tested by an indirect absorbed enzyme–linked immunosorbent assay (ELISA-C. All fecal samples were tested by pooled culture. After that, fecal samples of Map positive pools were tested individually by culture and polymerase chain reaction (PCR. In one herd, slurry and tissue samples from one animal were also taken and tested by PCR and culture. Map isolates were analyzed by the Multilocus Short Sequence Repeat (MLSSR and the Mycobacterial Interspersed Repetitive Units-Variable Number of Tandem Repeats (MIRU-VNTR methods. ELISA produced positive results in 1.8% (6/329 of the animals and 40% (2/5 of the herds. Four fecal, two tissue, and two slurry samples from a herd were Map positive by culture and PCR. MLSSR and MIRU-VNTR revealed two different strain profiles among eight Map isolates recovered. This study reports the first molecular characterization of Map in one dairy herd in Colombia, the limitations for individual diagnosis of subclinical Map infections in cattle, and the usefulness of pooled fecal samples and environmental sampling for Map diagnosis.

  13. Diagnosis and Molecular Characterization of Mycobacterium avium subsp. paratuberculosis from Dairy Cows in Colombia

    Science.gov (United States)

    Fernández-Silva, J. A.; Abdulmawjood, A.; Bülte, M.

    2011-01-01

    The objective of this study was the serological, bacteriological and molecular diagnosis, as well as the molecular characterization of Mycobacterium avium subsp. paratuberculosis (Map) in adult cows of five Colombian dairy herds. Serum samples were tested by an indirect absorbed enzyme–linked immunosorbent assay (ELISA-C). All fecal samples were tested by pooled culture. After that, fecal samples of Map positive pools were tested individually by culture and polymerase chain reaction (PCR). In one herd, slurry and tissue samples from one animal were also taken and tested by PCR and culture. Map isolates were analyzed by the Multilocus Short Sequence Repeat (MLSSR) and the Mycobacterial Interspersed Repetitive Units-Variable Number of Tandem Repeats (MIRU-VNTR) methods. ELISA produced positive results in 1.8% (6/329) of the animals and 40% (2/5) of the herds. Four fecal, two tissue, and two slurry samples from a herd were Map positive by culture and PCR. MLSSR and MIRU-VNTR revealed two different strain profiles among eight Map isolates recovered. This study reports the first molecular characterization of Map in one dairy herd in Colombia, the limitations for individual diagnosis of subclinical Map infections in cattle, and the usefulness of pooled fecal samples and environmental sampling for Map diagnosis. PMID:21785685

  14. Molecular characterization, isolation, pathology and pathotyping of peafowl (Pavo cristatus) origin Newcastle disease virus isolates recovered from disease outbreaks in three states of India.

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    Desingu, Perumal Arumugam; Singh, Shambhu Dayal; Dhama, Kuldeep; Vinodhkumar, Obli Rajendran; Barathidasan, Rajamani; Malik, Yashpal Singh; Singh, Rajendra; Singh, Raj Kumar

    2016-12-01

    Disease outbreak investigations were carried out in three states of Northern India namely Haryana (Rewari), Uttar Pradesh (Noida) and Delhi, where a total of 110 Indian peafowls (Pavo cristatus) showed sudden onset of nervous signs and died within a period of two weeks during June, 2012. The F (fusion) gene-based RT-PCR detection of Newcastle disease virus (NDV) in affected tissues confirmed the presence of the virus. Three NDV isolates were selected (one from each area under investigation) and further characterized. They were found to be of virulent pathotype (velogenic NDV) based on both pathogenicity assays (MDT, ICPI and IVPI) and partial F gene sequence analysis. Additionally, the phylogenetic analysis revealed that the isolates belonged to the genotype VIIi and XIII of class II avian Paramyxovirus serotype1 (APMV-1) and related closely to new emerging sub-genotypes. This is the first report regarding the presence of the fifth panzootic vNDV genotype VIIi from India. In this scenario, extensive epidemiological studies are suggested for surveillance of NDV genotypes in wild birds and poultry flocks of the country along with adopting suitable prevention and control measures.

  15. Characterization of chromosomal qnrB and ampC alleles in Citrobacter freundii isolates from different origins.

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    Liao, Xiaoping; Fang, Liangxing; Li, Liang; Sun, Jian; Li, Xingping; Chen, Muya; Deng, Hui; Yang, Qiu'e; Li, Xue; Liu, Yahong

    2015-10-01

    The association of ESBLs (extended-spectrum beta-lactamases)/pAmpCs (plasmid-mediated AmpC β-lactamases) with PMQR (plasmid mediated quinolone resistance) in gram-negative bacteria has been of great concern. The present study was performed to characterize the diversity, gene location, genetic context, and evolution of ampC and qnrB alleles in isolates of Citrobacter freundii. Fifteen isolates of C. freundii were identified from a total of 788 isolates of Enterobacteriaceae derived from humans, animals, animal food products, and the environment between 2010 and 2012. Co-existence of qnrB/ΔqnrB with ampC was detected in all C. freundii isolates. Both ampC and qnrB genes were found to be located on the chromosome, but were distantly separated on the chromosome. Seven and six novel alleles were discovered for the 10 ampC and qnrB variants detected in this study, respectively. Phylogenetic analysis showed that the new alleles differed a little from the variants of ampC/qnrB previously described in this genus. The genetic context surrounding ampC genes was AmpR-AmpC-Blc-SugE. However, five different genetic contexts surrounding qnrB/ΔqnrB genes were observed, but they occurred in all cases between the pspF and sapA genes. Additionally, cloning experiments showed that the regions containing different qnrB alleles, even with different genetic contexts, contributed to the reduction of quinolone susceptibility. Our results showed that the chromosomal ampC and qnrB alleles are closely related to C. freundii. However, unlike ampC, qnrB alleles seemed to be related to the genetic contexts surrounding them. The evolution of these two genes in C. freundii isolates might be through different pathways.

  16. Infection Sources of a Common Non-tuberculous Mycobacterial Pathogen, Mycobacterium avium Complex

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    Nishiuchi, Yukiko; Iwamoto, Tomotada; Maruyama, Fumito

    2017-01-01

    Numerous studies have revealed a continuous increase in the worldwide incidence and prevalence of non-tuberculous mycobacteria (NTM) diseases, especially pulmonary Mycobacterium avium complex (MAC) diseases. Although it is not clear why NTM diseases have been increasing, one possibility is an increase of mycobacterial infection sources in the environment. Thus, in this review, we focused on the infection sources of pathogenic NTM, especially MAC. The environmental niches for MAC include water, soil, and dust. The formation of aerosols containing NTM arising from shower water, soil, and pool water implies that these niches can be infection sources. Furthermore, genotyping has shown that clinical isolates are identical to environmental ones from household tap water, bathrooms, potting soil, and garden soil. Therefore, to prevent and treat MAC diseases, it is essential to identify the infection sources for these organisms, because patients with these diseases often suffer from reinfections and recurrent infections with them. In the environmental sources, MAC and other NTM organisms can form biofilms, survive within amoebae, and exist in a free-living state. Mycobacterial communities are also likely to occur in these infection sources in households. Water distribution systems are a transmission route from natural water reservoirs to household tap water. Other infection sources include areas with frequent human contact, such as soil and bathrooms, indicating that individuals may carry NTM organisms that concomitantly attach to their household belongings. To explore the mechanisms associated with the global spread of infection and MAC transmission routes, an epidemiological population-wide genotyping survey would be very useful. A good example of the power of genotyping comes from M. avium subsp. hominissuis, where close genetic relatedness was found between isolates of it from European patients and pigs in Japan and Europe, implying global transmission of this bacterium

  17. Antimicrobial Resistance Profiles of Campylobacter spp. Isolated from Broiler Chicken Meat of Estonian, Latvian and Lithuanian Origin at Estonian Retail Level and from Patients with Severe Enteric Infections in Estonia.

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    Mäesaar, M; Kramarenko, T; Meremäe, K; Sõgel, J; Lillenberg, M; Häkkinen, L; Ivanova, M; Kovalenko, K; Hörman, A; Hänninen, M-L; Roasto, M

    2016-03-01

    The resistance patterns of Campylobacter spp. isolated from retail broiler chicken meat originating either from Estonia, Lithuania or Latvia collected in Estonia were determined. Additionally, in collaboration with the laboratories of several Estonian hospitals, antimicrobial susceptibility patterns were determined for Campylobacter isolates from patients with severe Campylobacter enteric infections. The isolates were identified at the species level by the PCR method. Respectively, 88.8% of the isolates were C. jejuni, and 11.2% were C. coli. In total, 126 Campylobacter isolates of broiler chicken meat and human origin were tested for minimal inhibitory concentrations (MICs) with the broth microdilution VetMIC(TH) method (National Veterinary Institute; Uppsala, Sweden) for a total of six antimicrobials. Resistance to one or more antimicrobials was detected in 62 (63.3%) of Campylobacter broiler chicken meat isolates and in 20 (71.4%) of human-origin isolates. Large proportions of the broiler chicken meat isolates were resistant to ciprofloxacin (60.2%). Multidrug resistance (i.e. to three or more unrelated antimicrobials) was detected in five (5.1%) C. jejuni isolates. Among the human isolates, 20 (71.4%) were resistant to fluoroquinolones, and two (7.1%) C. jejuni isolates exhibited multidrug resistance. The chicken meat isolates of Estonian origin were the most susceptible. However, a high proportion of fluoroquinolone-resistant C. jejuni isolates were found in Latvian and Lithuanian products. The results of this study indicate that the problems caused by the inappropriate use of antimicrobials extend beyond the country in which a food originates; therefore, both domestic and international interventions and agreements are required to implement common policies on antimicrobial usage and to minimize the emergence of Campylobacter drug resistance.

  18. Evaluation of Leishmania (Leishmania chagasi strains isolated from dogs originating from two visceral leishmaniasis-endemic areas in Brazil using multilocus enzyme electrophoresis

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    Carlos Eduardo Ribeiro Coutinho

    2011-10-01

    Full Text Available INTRODUCTION: Domestic dogs are the most important reservoir in the peridomestic transmission cycle of Leishmania (Leishmania chagasi. The genetic variability of subpopulations of this parasite circulating in dogs has not been thoroughly analyzed in Brazil, even though this knowledge has important implications in the clinical-epidemiological context. METHODS: The objective of this study was to evaluate and compare the phenotypic variability of 153 L. chagasi strains isolated from dogs originating from the municipalities of Rio de Janeiro (n = 57 and Belo Horizonte (n = 96, where the disease is endemic. Strains isolated only from intact skin were selected and analyzed by multilocus enzyme electrophoresis using nine enzyme systems (6PG, GPI, NH1 and NH2, G6P, PGM, MDH, ME, and IDHNADP. RESULTS: The electrophoretic profile was identical for all isolates analyzed and was the same as that of the L. chagasi reference strain (MHOM/BR/74/PP75. Phenetic analysis showed a similarity index of one for all strains, with the isolates sharing 100% of the characteristics analyzed. CONCLUSIONS: The results demonstrate that the L. chagasi populations circulating in dogs from Rio de Janeiro and Belo Horizonte belong to a single zymodeme.

  19. Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) isolates of swine origin form robust biofilms.

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    Nicholson, Tracy L; Shore, Sarah M; Smith, Tara C; Frana, Timothy S; Fraena, Timothy S

    2013-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) colonization of livestock animals is common and prevalence rates for pigs have been reported to be as high as 49%. Mechanisms contributing to the persistent carriage and high prevalence rates of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) strains in swine herds and production facilities have not been investigated. One explanation for the high prevalence of MRSA in swine herds is the ability of these organisms to exist as biofilms. In this report, the ability of swine LA-MRSA strains, including ST398, ST9, and ST5, to form biofilms was quantified and compared to several swine and human isolates. The contribution of known biofilm matrix components, polysaccharides, proteins and extracellular DNA (eDNA), was tested in all strains as well. All MRSA swine isolates formed robust biofilms similar to human clinical isolates. The addition of Dispersin B had no inhibitory effect on swine MRSA isolates when added at the initiation of biofilm growth or after pre-established mature biofilms formed. In contrast, the addition of proteinase K inhibited biofilm formation in all strains when added at the initiation of biofilm growth and was able to disperse pre-established mature biofilms. Of the LA-MRSA strains tested, we found ST398 strains to be the most sensitive to both inhibition of biofilm formation and dispersal of pre-formed biofilms by DNaseI. Collectively, these findings provide a critical first step in designing strategies to control or eliminate MRSA in swine herds.

  20. Molecular characterization of African swine fever virus isolates originating from outbreaks in the Russian Federation between 2007 and 2011.

    Science.gov (United States)

    Malogolovkin, Alexander; Yelsukova, Alexandra; Gallardo, Carmina; Tsybanov, Sodnom; Kolbasov, Denis

    2012-08-17

    African swine fever is one of the most important viral diseases of pigs and which caused significant economic damage on the pig production worldwide. Nowadays, it is still present on the African continent, in Transcaucasus countries (TCC), on Island of Sardinia and in Russia. Outbreaks of the disease have been reported in Russia for the last four years, affected especially the Southern Federal District of the country. Since 2010, a new outbreak area has been observed in the Northwestern Federal District. In order to study the evolution of African swine fever virus (ASFV) isolates, strains were collected in the Russian Federation from 2007 to 2011 and investigated by means of partial sequencing and fragment length polymorphism. In detail, 7 variable regions, namely B646L, E183L, I196L, B602L, I73R/I329R, I78R/I215L and KP86R were investigated. Phylogenetic analyses revealed 100% nucleotide identity of B646L and E183L gene sequences of all examined isolates. All isolates formed one genetic cluster within genotype II. Moreover, no amplified fragment length polymorphism (AFLP) was observed for B602L, I196L, I73R/I329R, and I78R/I215L genes. The flanking primers used to amplify the KP86R gene failed to amplify a product in all the isolates. The obtained data strongly suggests that only one ASFV virus variant caused the outbreaks from 2007 to 2011 in the territory of the Russian Federation.

  1. Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA isolates of swine origin form robust biofilms.

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    Tracy L Nicholson

    Full Text Available Methicillin-resistant Staphylococcus aureus (MRSA colonization of livestock animals is common and prevalence rates for pigs have been reported to be as high as 49%. Mechanisms contributing to the persistent carriage and high prevalence rates of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA strains in swine herds and production facilities have not been investigated. One explanation for the high prevalence of MRSA in swine herds is the ability of these organisms to exist as biofilms. In this report, the ability of swine LA-MRSA strains, including ST398, ST9, and ST5, to form biofilms was quantified and compared to several swine and human isolates. The contribution of known biofilm matrix components, polysaccharides, proteins and extracellular DNA (eDNA, was tested in all strains as well. All MRSA swine isolates formed robust biofilms similar to human clinical isolates. The addition of Dispersin B had no inhibitory effect on swine MRSA isolates when added at the initiation of biofilm growth or after pre-established mature biofilms formed. In contrast, the addition of proteinase K inhibited biofilm formation in all strains when added at the initiation of biofilm growth and was able to disperse pre-established mature biofilms. Of the LA-MRSA strains tested, we found ST398 strains to be the most sensitive to both inhibition of biofilm formation and dispersal of pre-formed biofilms by DNaseI. Collectively, these findings provide a critical first step in designing strategies to control or eliminate MRSA in swine herds.

  2. Genetic relatedness of Plasmodium falciparum isolates and the origin of allelic diversity at the merozoite surface protein-1 (MSP-1 locus in Brazil and Vietnam

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    Ferreira Marcelo U

    2003-07-01

    Full Text Available Abstract Background Despite the extensive polymorphism at the merozoite surface protein-1 (MSP-1 locus of Plasmodium falciparum, that encodes a major repetitive malaria vaccine candidate antigen, identical and nearly identical alleles frequently occur in sympatric parasites. Here we used microsatellite haplotyping to estimate the genetic distance between isolates carrying identical and nearly identical MSP-1 alleles. Methods We analyzed 28 isolates from hypoendemic areas in north-western Brazil, collected between 1985 and 1998, and 23 isolates obtained in mesoendemic southern Vietnam in 1996. MSP-1 alleles were characterized by combining PCR typing with allele-specific primers and partial DNA sequencing. The following single-copy microsatellite markers were typed : Polyα, TA42 (only for Brazilian samples, TA81, TA1, TA87, TA109 (only for Brazilian samples, 2490, ARAII, PfG377, PfPK2, and TA60. Results The low pair-wise average genetic distance between microsatellite haplotypes of isolates sharing identical MSP-1 alleles indicates that epidemic propagation of discrete parasite clones originated most identical MSP-1 alleles in parasite populations from Brazil and Vietnam. At least one epidemic clone propagating in Brazil remained relatively unchanged over more than one decade. Moreover, we found no evidence that rearrangements of MSP-1 repeats, putatively created by mitotic recombination events, generated new alleles within clonal lineages of parasites in either country. Conclusion Identical MSP-1 alleles originated from co-ancestry in both populations, whereas nearly identical MSP-1 alleles have probably appeared independently in unrelated parasite lineages.

  3. Phylogenetic analysis of NS5B gene of classical swine fever virus isolates indicates plausible Chinese origin of Indian subgroup 2.2 viruses.

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    Patil, S S; Hemadri, D; Veeresh, H; Sreekala, K; Gajendragad, M R; Prabhudas, K

    2012-02-01

    Twenty-three CSFV isolates recovered from field outbreaks in various parts of India during 2006-2009 were used for genetic analysis in the NS5B region (409 nts). Seventeen of these were studied earlier [16] in the 5'UTR region. Phylogenetic analysis indicated the continued dominance of subgroup 1.1 strains in the country. Detailed analysis of a subgroup 2.2 virus indicated the plausible Chinese origin of this subgroup in India and provided indirect evidence of routes of CSFV movement within South East Asia region.

  4. Mycobacterium avium subspecies induce differential expression of pro-inflammatory mediators in a murine macrophage model: evidence for enhanced pathogenicity of Mycobacterium avium subspecies paratuberculosis.

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    Basler, Tina; Geffers, Robert; Weiss, Siegfried; Valentin-Weigand, Peter; Goethe, Ralph

    2008-01-01

    Mycobacterium avium subspecies (ssp.) paratuberculosis (MAP) is the etiological agent of paratuberculosis, a chronic, non-treatable granulomatous enteritis of ruminants. MAP is the only mycobacterium affecting the intestinal tract, which is of interest since it is presently the most favoured pathogen linked to Crohn's disease (CD) in humans due to its frequent detection in CD tissues. MAP is genetically closely related to other M. avium ssp. such as M. avium ssp. avium (MAA) and M. avium ssp. hominissuis (MAH) which can cause mycobacteriosis in animals and immunocompromised humans. We have recently shown that murine macrophage cell lines represent suitable systems to analyse M. avium ssp. patho-mechanisms and could show that MAP, but not MAA, specifically inhibited the antigen-specific stimulatory capacity for CD4(+) T-cells. In the present study, we compared gene expression profiles of murine RAW264.7 macrophages in response to infections with MAP or MAA using murine high-density oligonucleotide Affymetrix microarrays. A comparison of MAP and MAA infection revealed 17 differentially expressed genes. They were expressed at a much lower level in MAP-infected macrophages than in MAA-infected macrophages. Among these were the genes for IL-1beta, IL-1alpha, CXCL2, PTGS2 (COX2), lipocalin (LCN2) and TNF, which are important pro-inflammatory factors. The microarray data were confirmed for selected genes by quantitative real-time reverse transcription PCR and, by protein array analyses and ELISA. Similar to MAA, infection with MAH also showed robust induction of IL-1beta, CXCL2, COX2, LCN2 and TNF. Taken together, our results from M. avium ssp.-infected murine macrophages provide evidence that MAP in contrast to MAA and MAH specifically suppresses the pro-inflammatory defence mechanisms of infected macrophages.

  5. Detection of Mycobacterium avium subspecies paratuberculosis of dairy cows in Bogor

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    Widagdo Sri Nugroho

    2009-12-01

    Full Text Available Johne’s disease (JD or partuberculosis is a chronic granulomatous enteritis in ruminants caused by infection of Mycobacterium avium paratuberculosis subspecies (MAP. The disease has been detected serologically in Indonesia. It’s potential to spread to other herds and could create great economic losses. The objectives of current study were to detect MAP in milk and faeces of dairy cows as well as to evaluate the association between farm management factors and presence of the bacteria in dairy cows in Bogor. The sample size was calculated using the formula to detect disease with the prevalence assumed to be 5% using 95% significant level. Milk and faeces samples were taken from 62 dairy cows which were suspected as suffering from MAP infection. Detection of MAP was done by isolation in Herrold’ egg yolk medium with mycobactin J (HEYMj, acid-fast bacilli Ziehl-Neelsen staining, PCR IS900 and F57. Biochemical test to confirm M. tuberculosis presence was also conducted. Fifteen isolates of Mycobacterium sp. were found from the faeces samples but not from the corresponding milk samples. However, conventional PCR conducted on the isolate as well as the milk samples, gave negative results. Biochemical test proved that all Mycobacterium sp. isolates were not M. tuberculosis. This study indicated the prevalence of MAP in Bogor was less than 5%. These findings should be continued by observational study to achieve the comprehensive information at the cattle and herd level. Bovine Tuberculosis monitoring should be done also to protect dairy herd and food safety for the community.

  6. Isolation of Novel Trypanosomatid, Zelonia australiensis sp. nov. (Kinetoplastida: Trypanosomatidae) Provides Support for a Gondwanan Origin of Dixenous Parasitism in the Leishmaniinae

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    Barratt, Joel; Kaufer, Alexa; Peters, Bryce; Craig, Douglas; Lawrence, Andrea; Roberts, Tamalee; Lee, Rogan; McAuliffe, Gary; Stark, Damien; Ellis, John

    2017-01-01

    The genus Leishmania includes approximately 53 species, 20 of which cause human leishmaniais; a significant albeit neglected tropical disease. Leishmaniasis has afflicted humans for millennia, but how ancient is Leishmania and where did it arise? These questions have been hotly debated for decades and several theories have been proposed. One theory suggests Leishmania originated in the Palearctic, and dispersed to the New World via the Bering land bridge. Others propose that Leishmania evolved in the Neotropics. The Multiple Origins theory suggests that separation of certain Old World and New World species occurred due to the opening of the Atlantic Ocean. Some suggest that the ancestor of the dixenous genera Leishmania, Endotrypanum and Porcisia evolved on Gondwana between 90 and 140 million years ago. In the present study a detailed molecular and morphological characterisation was performed on a novel Australian trypanosomatid following its isolation in Australia’s tropics from the native black fly, Simulium (Morops) dycei Colbo, 1976. Phylogenetic analyses were conducted and confirmed this parasite as a sibling to Zelonia costaricensis, a close relative of Leishmania previously isolated from a reduviid bug in Costa Rica. Consequently, this parasite was assigned the name Zelonia australiensis sp. nov. Assuming Z. costaricensis and Z. australiensis diverged when Australia and South America became completely separated, their divergence occurred between 36 and 41 million years ago at least. Using this vicariance event as a calibration point for a phylogenetic time tree, the common ancestor of the dixenous genera Leishmania, Endotrypanum and Porcisia appeared in Gondwana approximately 91 million years ago. Ultimately, this study contributes to our understanding of trypanosomatid diversity, and of Leishmania origins by providing support for a Gondwanan origin of dixenous parasitism in the Leishmaniinae. PMID:28081121

  7. Surviving within the amoebal exocyst: the Mycobacterium avium complex paradigm

    Directory of Open Access Journals (Sweden)

    Drancourt Michel

    2010-04-01

    Full Text Available Abstract Background Most of environmental mycobacteria have been previously demonstrated to resist free-living amoeba with subsequent increased virulence and resistance to antibiotics and biocides. The Mycobacterium avium complex (MAC comprises of environmental organisms that inhabit a wide variety of ecological niches and exhibit a significant degree of genetic variability. We herein studied the intra-ameobal location of all members of the MAC as model organisms for environmental mycobacteria. Results Type strains for M. avium, Mycobacterium intracellulare, Mycobacterium chimaera, Mycobacterium colombiense, Mycobacterium arosiense, Mycobacterium marseillense, Mycobacterium timonense and Mycobacterium bouchedurhonense were co-cultivated with the free-living amoeba Acanthamoeba polyphaga strain Linc-AP1. Microscopic analyses demonstrated the engulfment and replication of mycobacteria into vacuoles of A. polyphaga trophozoites. Mycobacteria were further entrapped within amoebal cysts, and survived encystment as demonstrated by subculturing. Electron microscopy observations show that, three days after entrapment into A. polyphaga cysts, all MAC members typically resided within the exocyst. Conclusions Combined with published data, these observations indicate that mycobacteria are unique among amoeba-resistant bacteria, in residing within the exocyst.

  8. Iodixanol Development of a Laboratory-Scale Technique to Monitor the Persistence of Mycobacterium avium subsp. paratuberculosis in Cheddar Cheese

    Directory of Open Access Journals (Sweden)

    J. A. Donaghy

    2003-01-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (Map is a potential human pathogen known to be present in raw milk from infected dairy herds. Current pasteurisation regimes do not totally inactivate Map resulting in the possibility of viable cells being present in pasteurised milk used for Cheddar cheese production. A laboratory-based method, ensuring strict safety precautions, was developed to manufacture 800-g Cheddar blocks, experimentally contaminated (postpasteurisation with two different strains of Map. The composition of the model Cheddar produced was consistent with commercial product. Syneresis of the cheese curd caused a 1 log10 concentration of Map numbers from milk to cheese for a strain isolated from pasteurised milk. The type strain NCTC 8578 did not show a similar concentration effect, but did however survive the Cheddar manufacturing process. A small percentage (<5% of the Map load for each strain was recovered in the whey fraction during the process.

  9. The white morphotype of Mycobacterium avium-intracellulare is common in infected humans and virulent in infection models.

    Science.gov (United States)

    Mukherjee, S; Petrofsky, M; Yaraei, K; Bermudez, L E; Cangelosi, G A

    2001-12-01

    Isolates of Mycobacterium avium-intracellulare (MAI) form multiple colony types named red-opaque, white-opaque, red-transparent (RT), and white-transparent (WT). The newly discovered WT morphotype is multidrug resistant relative to other variants in vitro. To determine whether the WT morphotype occurs in humans, 32 MAI-positive clinical samples from 2 sites were plated directly onto indicator agar without prior passage in vitro. WT was the predominant morphotype in 26 (81%) of these samples and was absent in only 2 samples. WT variants grew better than isogenic RT variants in mouse and human macrophage models of infection, and RT clones that passed through such systems underwent rapid shifts to the WT morphotype. The RT morphotype was heterogeneous with regard to infectivity. In summary, the white morphotype was common in humans and was favored in disease models. It may play an important role in the establishment and persistence of MAI infection.

  10. ORIGINAL ARTICLE: Antifungal Activity of Some Natural Essential Oils against Candida Species Isolated from Blood Stream Infection

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    Amit Kumar

    2012-01-01

    Full Text Available Background: Candida is a part of normal microflora of human body and exists as an opportunistic pathogen as it attacks immunocompromised patients. Aims and Objectives: Candida is the most commonly isolated organism from blood stream infections. Fluconazole is the major antifungal drug used for treatment of Candida. Resistance to fluconazole has been increasing in recent years so there should be search of some other alternative. To find out these alternatives, anti candidial activity of some natural essential oils was studied. Materials and Methods: In the present study nine Candida strains isolated from blood stream infections were collected from National Culture Collection of Pathogenic Fungi (N.C.C.P.F. P.G.I.M.E.R Chandigarh India and the antifungal activity of some natural essential oils such as lemongrass oil, coconut oil, almond oil and clove oil was checked by using agar diffusion method. Result: All oils have shown a significantly anti-candidal activity. However, the antifungal activity was maximum in lemongrass oil. Conclusion: Our study may help to design new chemotherapeutic strategies against Candidal infections.

  11. Modeling the effect of direct and indirect contamination of on-farm bulk tank milk with Mycobacterium avium subsp. paratuberculosis

    DEFF Research Database (Denmark)

    Okura, Hisako; Nielsen, Søren Saxmose; Toft, Nils

    2013-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) in milk of bovine origin is suspected of being implicated in Crohn's disease in humans. Milk can be contaminated via direct excretion of MAP in milk or indirectly via fecal contamination of the milk. This study aimed at estimating the level of MAP...... milk from test positive cows would result in discarding 11% of milk and reduce the MAP level by 80%. Due to poor sensitivity of the diagnostic test, removing test-positive cows would not further reduce the already low concentration of MAP and it would not guarantee the milk as MAP-free. The model...

  12. Epidemiology and Ecology of Opportunistic Premise Plumbing Pathogens: Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa

    Science.gov (United States)

    BACKGROUND: Legionella pneumophila, Mycobacterium avium, and Pseudomonas aeruginosa are opportunistic premise plumbing pathogens (OPPPs) that persist and grow in household plumbing, habitats they share with humans. Infections caused by these OPPPs involve individuals with preexis...

  13. New genetic evidence supports isolation and drift in the Ladin communities of the South Tyrolean Alps but not an ancient origin in the Middle East.

    Science.gov (United States)

    Thomas, Mark G; Barnes, Ian; Weale, Michael E; Jones, Abigail L; Forster, Peter; Bradman, Neil; Pramstaller, Peter P

    2008-01-01

    The Alps are one of the most significant geographical barriers in Europe and several isolated Swiss and Italian valleys retain the distinctive Ladin and Romansch languages, alongside the modern majority of Italian and German languages. Linguistically, Ladin belongs to the Romance languages, but some studies on mitochondrial DNA (mtDNA) variation have suggested a major Middle Eastern component to their genealogical origin. Furthermore, an observed high degree of within-population diversity has been interpreted as reflecting long-standing differentiation from other European populations and the absence of a major bottleneck in Ladin population history. To explore these issues further, we examined Y chromosome and mtDNA variation in two samples of Ladin speakers, two samples of German speakers and one sample of metropolitan Italian speakers. Our results (1) indicate reduced diversity in the Ladin-speaking and isolated German-speaking populations when compared to a sample of metropolitan Italian speakers, (2) fail to identify haplotypes that are rare in other European populations that other researchers have identified, and (3) indicate different Middle Eastern components to Ladin ancestry in different localities. These new results, in combination with Bayesian estimation of demographic parameters of interest (population size, population growth rate, and Palaeolithic/Neolithic admixture proportions) and phylogeographic analysis, suggest that the Ladin groups under study are small genetically isolated populations (subject to strong genetic drift), having a predominantly European ancestry, and in one locality, may have a greater Palaeolithic component to that ancestry than their neighbours.

  14. Effect of lipoarabinomannan from Mycobacterium avium subsp avium in Freund’s incomplete adjuvant on the immune response of cattle

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    S.B. Colavecchia

    2012-02-01

    Full Text Available The aim of the present study was to determine whether lipoarabinomannan (LAM, in combination with Freund’s incomplete adjuvant (FIA, was able to improve cell-mediated and antibody-mediated immune responses against ovalbumin (OVA in cattle. Twenty-three calves were assigned to four treatment groups, which were subcutaneously immunized with either OVA plus FIA, OVA plus FIA and LAM from Mycobacterium avium subsp avium, FIA plus LAM, or FIA alone. Lymphoproliferation, IFN-γ production and cell subpopulations on peripheral blood mononuclear cells before and 15 days after treatment were evaluated. Delayed hypersensitivity was evaluated on day 57. Specific humoral immune response was measured by ELISA. Inoculation with LAM induced higher levels of lymphoproliferation and IFN-γ production in response to ConA and OVA (P < 0.05. Specific antibody titers were similar in both OVA-immunized groups. Interestingly, our results showed that the use of LAM in vaccine preparations improved specific cell immune response evaluated by lymphoproliferation and IFN-γ production by at least 50 and 25%, respectively, in cattle without interfering with tuberculosis and paratuberculosis diagnosis.

  15. Mycobacterium avium subsp. paratuberculosis is not discerned in diabetes mellitus patients in Hyderabad, India.

    Science.gov (United States)

    Rani, Pittu Sandhya; Doddam, Sankara Narayana; Agrawal, Sashank; Hasnain, Seyed E; Sechi, Leonardo A; Kumar, Ashutosh; Ahmed, Niyaz

    2014-07-01

    Mycobacterium avium ssp. paratuberculosis (MAP) is an obligate intracellular pathogen. It causes chronic intestinal inflammation in ruminants known as Johne's disease and is associated with human Crohn's disease. Furthermore, association of MAP with other autoimmune diseases, such as type-1 diabetes, has been established in patients from Sardinia (Italy) which is a MAP endemic and genetically isolated region. Due to largest livestock population and consequently high MAP prevalence amidst a very high diabetes incidence in India, we sought to test this association on a limited number of patient samples from Hyderabad. Our results of ELISA with MAP lysate and MAP-specific protein MAP3738c as well as PCR/real-time PCR of MAP-specific sequences IS900 and/or f57 indicated that, in contrast to Sardinian diabetic patients, MAP infection in blood is not discerned in diabetic patients in Hyderabad. The association of a mycobacterial trigger with diabetes therefore could well be a population-specific phenomenon, highly dependent on genetic repertoire and the environment of susceptible populations. However, a larger study is needed in order to confirm this.

  16. LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis.

    Science.gov (United States)

    Trangoni, Marcos D; Gioffré, Andrea K; Cerón Cucchi, María E; Caimi, Karina C; Ruybal, Paula; Zumárraga, Martín J; Cravero, Silvio L

    2015-06-01

    In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR Green(TM) allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries.

  17. Bordetella avium Causes Induction of Apoptosis and Nitric Oxide Synthase in Turkey Tracheal Explant Cultures

    OpenAIRE

    Miyamoto, David M.; Ruff, Kristin; Beach, Nathan M.; Dorsey-Oresto, Angella; Masters, Isaac; Temple, Louise M.

    2011-01-01

    Bordetellosis is an upper respiratory disease of turkeys caused by Bordetella avium in which the bacteria attach specifically to ciliated respiratory epithelial cells. Little is known about the mechanisms of pathogenesis of this disease, which has a negative impact in the commercial turkey industry. In this study, we produced a novel explant organ culture system that was able to successfully reproduce pathogenesis of B. avium in vitro, using tracheal tissue derived from 26 day-old turkey embr...

  18. Infection by Mycobacterium avium intracellulare in AIDS: endoscopic duodenal appearance mimicking Whipple's disease.

    Science.gov (United States)

    Vázquez-Iglesias, J L; Yañez, J; Durana, J; Arnal, F

    1988-09-01

    We report the case of a 24-year-old woman who presented with diarrhea, weight loss and abdominal lymph node enlargement. A diagnosis of infection by Mycobacterium avium intracellulare with a clinical picture similar to Whipple's disease was established. The endoscopic study of the duodenum revealed multiple yellow nodules that became confluent in the second portion, entirely replacing the normal mucosa. These endoscopic findings have not been described previously in intestinal infection by Mycobacterium avium intracellulare.

  19. Maximum growth rate of Mycobacterium avium in continuous culture or chronically infected BALB/c mice.

    Science.gov (United States)

    McCarthy, C M; Taylor, M A; Dennis, M W

    1987-01-01

    Mycobacterium avium is a human pathogen which may cause either chronic or disseminated disease and the organism exhibits a slow rate of growth. This study provides information on the growth rate of the organism in chronically infected mice and its maximal growth rate in vitro. M. avium was grown in continuous culture, limited for nitrogen with 0.5 mM ammonium chloride and dilution rates that ranged from 0.054 to 0.153 h-1. The steady-state concentration of ammonia nitrogen and M. avium cells for each dilution rate were determined. The bacterial saturation constant for growth-limiting ammonia was 0.29 mM (4 micrograms nitrogen/ml) and, from this, the maximal growth rate for M. avium was estimated to be 0.206 h-1 or a doubling time of 3.4 h. BALB/c mice were infected intravenously with 3 x 10(6) colony-forming units and a chronic infection resulted, typical of virulent M. avium strains. During a period of 3 months, the number of mycobacteria remained constant in the lungs, but increased 30-fold and 8,900-fold, respectively, in the spleen and mesenteric lymph nodes. The latter increase appeared to be due to proliferation in situ. The generation time of M. avium in the mesenteric lymph nodes was estimated to be 7 days.

  20. Expression library immunization confers protection against Mycobacterium avium subsp. paratuberculosis infection.

    Science.gov (United States)

    Huntley, J F; Stabel, J R; Paustian, M L; Reinhardt, T A; Bannantine, J P

    2005-10-01

    Currently, paratuberculosis vaccines are comprised of crude whole-cell preparations of Mycobacterium avium subsp. paratuberculosis. Although effective in reducing clinical disease and fecal shedding, these vaccines have severe disadvantages as well, including seroconversion of vaccinated animals and granulomatous lesions at the site of vaccination. DNA vaccines can offer an alternative approach that may be safer and elicit more protective responses. In an effort to identify protective M. avium subsp. paratuberculosis sequences, a genomic DNA expression library was generated and subdivided into pools of clones (approximately 1,500 clones/pool). The clone pools were evaluated to determine DNA vaccine efficacy by immunizing mice via gene gun delivery and challenging them with live, virulent M. avium subsp. paratuberculosis. Four clone pools resulted in a significant reduction in the amount of M. avium subsp. paratuberculosis recovered from mouse tissues compared to mice immunized with other clone pools and nonvaccinated, infected control mice. One of the protective clone pools was further partitioned into 10 clone arrays of 108 clones each, and four clone arrays provided significant protection from both spleen and mesenteric lymph node colonization by M. avium subsp. paratuberculosis. The nucleotide sequence of each clone present in the protective pools was determined, and coding region functions were predicted by computer analysis. Comparison of the protective clone array sequences implicated 26 antigens that may be responsible for protection in mice. This study is the first study to demonstrate protection against M. avium subsp. paratuberculosis infection with expression library immunization.

  1. Polyphyletic origin of MERS coronaviruses and isolation of a novel clade A strain from dromedary camels in the United Arab Emirates

    Science.gov (United States)

    Lau, Susanna K P; Wernery, Renate; Wong, Emily Y M; Joseph, Sunitha; Tsang, Alan K L; Patteril, Nissy Annie Georgy; Elizabeth, Shyna K; Chan, Kwok-Hung; Muhammed, Rubeena; Kinne, Jöerg; Yuen, Kwok-Yung; Wernery, Ulrich; Woo, Patrick C Y

    2016-01-01

    Little is known regarding the molecular epidemiology of Middle East respiratory syndrome coronavirus (MERS-CoV) circulating in dromedaries outside Saudi Arabia. To address this knowledge gap, we sequenced 10 complete genomes of MERS-CoVs isolated from 2 live and 8 dead dromedaries from different regions in the United Arab Emirates (UAE). Phylogenetic analysis revealed one novel clade A strain, the first detected in the UAE, and nine clade B strains. Strain D998/15 had a distinct phylogenetic position within clade A, being more closely related to the dromedary isolate NRCE-HKU205 from Egypt than to the human isolates EMC/2012 and Jordan-N3/2012. A comparison of predicted protein sequences also demonstrated the existence of two clade A lineages with unique amino acid substitutions, A1 (EMC/2012 and Jordan-N3/2012) and A2 (D998/15 and NRCE-HKU205), circulating in humans and camels, respectively. The nine clade B isolates belong to three distinct lineages: B1, B3 and B5. Two B3 strains, D1271/15 and D1189.1/15, showed evidence of recombination between lineages B4 and B5 in ORF1ab. Molecular clock analysis dated the time of the most recent common ancestor (tMRCA) of clade A to March 2011 and that of clade B to November 2011. Our data support a polyphyletic origin of MERS-CoV in dromedaries and the co-circulation of diverse MERS-CoVs including recombinant strains in the UAE. PMID:27999424

  2. Phosphoenolpyruvate carboxykinase in cherry (Prunus avium L.) fruit during development.

    Science.gov (United States)

    Walker, Robert P; Battistelli, Alberto; Moscatello, Stefano; Chen, Zhi-Hui; Leegood, Richard C; Famiani, Franco

    2011-11-01

    In this study the abundance and location of phosphoenolpyruvate carboxykinase (PEPCK) was determined in the flesh and skin of the sweet cherry (Prunus avium L.) cultivar Durone Nero II during development. PEPCK was not present in young fruit but appeared in both tissues as the fruit increased in size. In these there was no net dissimilation of malic acid, which accounts for the bulk of their organic acid contents when PEPCK was present. To assist in understanding the function of PEPCK, the abundance of a number of other enzymes was determined. These enzymes were aspartate aminotransferase (AspAT), glutamine synthetase (GS), phosphoenolpyruvate carboxylase (PEPC), pyruvate, orthophosphate dikinase (PPDK), and ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco). A potential role for PEPCK in the regulation of pH and the utilization of malate in gluconeogenesis in the flesh and skin of cherries is presented.

  3. Influence of cultivar and processing on cherry (Prunus avium) allergenicity.

    Science.gov (United States)

    Primavesi, L; Brenna, O V; Pompei, C; Pravettoni, V; Farioli, L; Pastorello, E A

    2006-12-27

    Oral allergy syndrome is an immediate food allergic event that affects lips, mouth, and pharynx, is often triggered by fruits and vegetables, and may be associated with pollinosis. Here, we report on the allergenic pattern of different varieties of cherry (Prunus avium) and results obtained by applying several technological processes to the selected varieties. Whole cherries were submitted to chemical peeling, thermal treatment, and syruping processes, and the relative protein extracts were analyzed by in vitro (sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting analysis) and in vivo tests (skin prick test). Electrophoretic analyses demonstrated that there was no marked difference among cherry cultivars. Chemical peeling successfully removed Pru av 3, a lipid transfer protein (LTP) responsible for oral allergy syndrome in patients without pollinosis, leading to the industrial production of cherry hypoallergenic derivatives. Furthermore, the syruping process removed almost all allergenic proteins to whom patients with pollinosis are responsive. In vivo tests confirmed electrophoretic results.

  4. Disseminated Mycobacterium avium complex infection in an immunocompetent pregnant woman

    Directory of Open Access Journals (Sweden)

    Kim Woo

    2006-10-01

    Full Text Available Abstract Background Disseminated mycobacterium avium complex (MAC occurs mainly in immunocompromised hosts, which is associated with abnormal cellular immunity. Case presentation A 26-year-old pregnant woman presented with fever and general weakness. Miliary lung nodules were noted on chest X-ray. Under the impression of miliary tuberculosis, anti-tuberculosis medication was administered. However, the patient was not improved. Further work-up demonstrated MAC in the sputum and placenta. The patient was treated successfully with clarithromycin-based combination regimen. Conclusion This appears to be the first case of disseminated MAC in an otherwise healthy pregnant woman. Clinicians should be alert for the diagnosis of MAC infection in diverse clinical conditions.

  5. Evaluation of Mycobacterium avium subsp. paratuberculosis faecal culture protocols and media Avaliação de protocolos de cultivo fecal e meios para a cultura de Mycobacterium avium subsp. paratuberculosis

    Directory of Open Access Journals (Sweden)

    Paula Ristow

    2006-03-01

    Full Text Available Paratuberculosis is an important enteritis of ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map. The disease is officially considered exotic in Brazil, but recent serological surveys and the isolation of the agent suggest it may occur in our herds. The aim of this study was to evaluate three different formulations of Herrold's egg yolk agar with mycobactin J (HEYM and four faecal culture protocols considering their ability for Map growth as well as cost and ease of application. Three formulations of HEYM were inoculated with two suspensions of Map. Spiked faeces and naturally contaminated faecal samples were treated by the four faecal culture protocols. Centrifugation protocol and HEYM recommended by OIE showed the best results on the recovery of Map.A paratuberculose é uma importante enterite de ruminantes causada por Mycobacterium avium subsp. paratuberculosis (Map. A enfermidade é oficialmente considerada exótica no Brasil, mas inquéritos sorológicos recentes e o isolamento do agente etiológico sugerem que ela pode estar presente em nossos rebanhos. O objetivo do estudo foi avaliar três diferentes fórmulas do Ágar gema de ovo de Herrold suplementado com micobactina J (HEYM e quatro protocolos de cultura fecal quanto ao crescimento de Map, bem como custo e facilidade de implementação. Três fórmulas de HEYM foram inoculadas com duas suspensões de Map. Fezes contaminadas artificialmente e naturalmente com Map foram tratadas pelos quatro protocolos de cultura fecal. O protocolo da centrifugação e a fórmula de HEYM recomendada pela OIE demonstraram os melhores resultados quanto à recuperação de Map.

  6. A point mutation in the polymerase protein PB2 allows a reassortant H9N2 influenza isolate of wild-bird origin to replicate in human cells.

    Science.gov (United States)

    Hussein, Islam T.M.; Ma, Eric J.; Meixell, Brandt; Hill, Nichola J.; Lindberg, Mark S.; Albrecht , Randy A.; Bahl, Justin; Runstadler, Jonathan A.

    2016-01-01

    H9N2 influenza A viruses are on the list of potentially pandemic subtypes. Therefore, it is important to understand how genomic reassortment and genetic polymorphisms affect phenotypes of H9N2 viruses circulating in the wild bird reservoir. A comparative genetic analysis of North American H9N2 isolates of wild bird origin identified a naturally occurring reassortant virus containing gene segments derived from both North American and Eurasian lineage ancestors. The PB2 segment of this virus encodes 10 amino acid changes that distinguish it from other H9 strains circulating in North America. G590S, one of the 10 amino acid substitutions observed, was present in ~ 12% of H9 viruses worldwide. This mutation combined with R591 has been reported as a marker of pathogenicity for human pandemic 2009 H1N1 viruses. Screening by polymerase reporter assay of all the natural polymorphisms at these two positions identified G590/K591 and S590/K591 as the most active, with the highest polymerase activity recorded for the SK polymorphism. Rescued viruses containing these two polymorphic combinations replicated more efficiently in MDCK cells and they were the only ones tested that were capable of establishing productive infection in NHBE cells. A global analysis of all PB2 sequences identified the K591 signature in six viral HA/NA subtypes isolated from several hosts in seven geographic locations. Interestingly, introducing the K591 mutation into the PB2 of a human-adapted H3N2 virus did not affect its polymerase activity. Our findings demonstrate that a single point mutation in the PB2 of a low pathogenic H9N2 isolate could have a significant effect on viral phenotype and increase its propensity to infect mammals. However, this effect is not universal, warranting caution in interpreting point mutations without considering protein sequence context.

  7. Pollen-Mediated Gene Flow in Maize: Implications for Isolation Requirements and Coexistence in Mexico, the Center of Origin of Maize.

    Directory of Open Access Journals (Sweden)

    Baltazar M Baltazar

    Full Text Available Mexico, the center of origin of maize (Zea mays L., has taken actions to preserve the identity and diversity of maize landraces and wild relatives. Historically, spatial isolation has been used in seed production to maintain seed purity. Spatial isolation can also be a key component for a strategy to minimize pollen-mediated gene flow in Mexico between transgenic maize and sexually compatible plants of maize conventional hybrids, landraces, and wild relatives. The objective of this research was to generate field maize-to-maize outcrossing data to help guide coexistence discussions in Mexico. In this study, outcrossing rates were determined and modeled from eight locations in six northern states, which represent the most economically important areas for the cultivation of hybrid maize in Mexico. At each site, pollen source plots were planted with a yellow-kernel maize hybrid and surrounded by plots with a white-kernel conventional maize hybrid (pollen recipient of the same maturity. Outcrossing rates were then quantified by assessing the number of yellow kernels harvested from white-kernel hybrid plots. The highest outcrossing values were observed near the pollen source (12.9% at 1 m distance. The outcrossing levels declined sharply to 4.6, 2.7, 1.4, 1.0, 0.9, 0.5, and 0.5% as the distance from the pollen source increased to 2, 4, 8, 12, 16, 20, and 25 m, respectively. At distances beyond 20 m outcrossing values at all locations were below 1%. These trends are consistent with studies conducted in other world regions. The results suggest that coexistence measures that have been implemented in other geographies, such as spatial isolation, would be successful in Mexico to minimize transgenic maize pollen flow to conventional maize hybrids, landraces and wild relatives.

  8. Pollen-Mediated Gene Flow in Maize: Implications for Isolation Requirements and Coexistence in Mexico, the Center of Origin of Maize.

    Science.gov (United States)

    Baltazar, Baltazar M; Castro Espinoza, Luciano; Espinoza Banda, Armando; de la Fuente Martínez, Juan Manuel; Garzón Tiznado, José Antonio; González García, Juvencio; Gutiérrez, Marco Antonio; Guzmán Rodríguez, José Luis; Heredia Díaz, Oscar; Horak, Michael J; Madueño Martínez, Jesús Ignacio; Schapaugh, Adam W; Stojšin, Duška; Uribe Montes, Hugo Raúl; Zavala García, Francisco

    2015-01-01

    Mexico, the center of origin of maize (Zea mays L.), has taken actions to preserve the identity and diversity of maize landraces and wild relatives. Historically, spatial isolation has been used in seed production to maintain seed purity. Spatial isolation can also be a key component for a strategy to minimize pollen-mediated gene flow in Mexico between transgenic maize and sexually compatible plants of maize conventional hybrids, landraces, and wild relatives. The objective of this research was to generate field maize-to-maize outcrossing data to help guide coexistence discussions in Mexico. In this study, outcrossing rates were determined and modeled from eight locations in six northern states, which represent the most economically important areas for the cultivation of hybrid maize in Mexico. At each site, pollen source plots were planted with a yellow-kernel maize hybrid and surrounded by plots with a white-kernel conventional maize hybrid (pollen recipient) of the same maturity. Outcrossing rates were then quantified by assessing the number of yellow kernels harvested from white-kernel hybrid plots. The highest outcrossing values were observed near the pollen source (12.9% at 1 m distance). The outcrossing levels declined sharply to 4.6, 2.7, 1.4, 1.0, 0.9, 0.5, and 0.5% as the distance from the pollen source increased to 2, 4, 8, 12, 16, 20, and 25 m, respectively. At distances beyond 20 m outcrossing values at all locations were below 1%. These trends are consistent with studies conducted in other world regions. The results suggest that coexistence measures that have been implemented in other geographies, such as spatial isolation, would be successful in Mexico to minimize transgenic maize pollen flow to conventional maize hybrids, landraces and wild relatives.

  9. Isolation and identification of unique marker compounds from the Tasmanian poppy Papaver somniferum N. Implications for the identification of illicit heroin of Tasmanian origin.

    Science.gov (United States)

    Odell, Luke R; Skopec, Jana; McCluskey, Adam

    2008-03-05

    Tasmanian opium accounts for 25% of the world's legal supply of opium straw, and in 1998-99 sufficient numbers of flower pods (66,013) to manufacture ca 500 kg of heroin were stolen. Whilst the heroin signature program has been developed to determine the origin of heroin from other key producers, no such signature currently exists for Tasmanian derived heroin. Tasmanian poppies contain a unique alkaloid, oripavine, which is the source of 'marker' impurities in illicit heroin produced from Tasmanian poppy straw. Treatment of oripavine (500mg) under Thiboumery and Mohr heroin processing conditions, followed by simple evaporative workup afforded 613 mg of a dark orange residue, which upon extensive chromatographic purification yielded oripavine 3-acetate (2) 22 mg; 3-acetyl-N-acetyldesthebaine (3) 35 mg; 3-acetyl-6-methoxy-4,5-epoxyphenanthrene (4) 5.8 mg; 3,4-diacetyl-6-methoxyphenanthrene (5) 27 mg; and 3,4,6-methoxy-5-[2(N-methylacetamido)]ethylphenanthrene (6) 52 mg. Compounds (2-6) are derived from oripavine and are unique to heroin derived from the Tasmanian poppy Papaver somniferum N. Analysis of illicit heroin samples seized from Turkey, Pakistan, Columbia and Myanmar did not reveal any of the aforementioned marker compounds. We have, however, identified four of these marker compounds (3-6) in seized heroin samples from Australia suggesting that they are of Tasmanian origin. Complete details of the isolation and identification of these compounds are provided.

  10. Zoonotic aspects of Mycobacterium bovis and Mycobacterium avium-intracellulare complex (MAC).

    Science.gov (United States)

    Biet, Franck; Boschiroli, Maria Laura; Thorel, Marie Françoise; Guilloteau, Laurence A

    2005-01-01

    Pathogens that are transmitted between the environment, wildlife, livestock and humans represent major challenges for the protection of human and domestic animal health, the economic sustainability of agriculture, and the conservation of wildlife. Among such pathogens, the genus Mycobacterium is well represented by M. bovis, the etiological agent of bovine tuberculosis, M. avium ssp. paratuberculosis (Map) the etiological agent of Johne disease, M. avium ssp. avium (Maa) and in a few common cases by other emergent environmental mycobacteria. Epidemiologic surveys performed in Europe, North America and New Zealand have demonstrated the existence and importance of environmental and wildlife reservoirs of mycobacterial infections that limit the attempts of disease control programmes. The aim of this review is to examine the zoonotic aspects of mycobacteria transmitted from the environment and wildlife. This work is focused on the species of two main groups of mycobacteria classified as important pathogens for humans and animals: first, M. bovis, the causative agent of bovine tuberculosis, which belongs to the M. tuberculosis complex and has a broad host range including wildlife, captive wildlife, domestic livestock, non-human primates and humans; the second group examined, is the M. avium-intracellulare complex (MAC) which includes M. avium ssp. avium causing major health problems in AIDS patients and M. avium ssp. paratuberculosis the etiological agent of Johne disease in cattle and identified in patients with Crohn disease. MAC agents, in addition to a broad host range, are environmental mycobacteria found in numerous biotopes including the soil, water, aerosols, protozoa, deep litter and fresh tropical vegetation. This review examines the possible reservoirs of these pathogens in the environment and in wildlife, their role as sources of infection in humans and animals and their health impact on humans. The possibilities of control and management programmes for

  11. Relationship between Presence of Cows with Milk Positive for Mycobacterium avium subsp. paratuberculosis-Specific Antibody by Enzyme-Linked Immunosorbent Assay and Viable M. avium subsp. paratuberculosis in Dust in Cattle Barns

    NARCIS (Netherlands)

    Eisenberg, S.W.F.; Chuchaisangrat, R.; Nielen, M.; Koets, A.P.

    2013-01-01

    Paratuberculosis, or Johne’s disease, in cattle is caused by Mycobacterium avium subsp. paratuberculosis, which has recently been suspected to be transmitted through dust. This longitudinal study on eight commercial M. avium subsp. paratuberculosispositive dairy farms studied the relationship betwee

  12. Concomitant Mycobacterium avium infection and Hodgkin's disease in a lymph node from an HIV-negative child.

    Science.gov (United States)

    de Armas, Yaxsier; Capó, Virginia; González, Ida; Mederos, Lilian; Díaz, Raúl; de Waard, Jacobus H; Rodríguez, Alberto; García, Yarmila; Cabanas, Ricardo

    2011-03-01

    We report a case of an immunocompetent child with simultaneously an infection with Mycobacterium avium and Hodgkin's disease in a cervical lymph node. A positive PCR result for M. avium on a biopsy of the lymph node directed the definitive diagnosis for both etiologies and avoided a possible dissemination of this infection after chemotherapy was started.

  13. Assessing the effectiveness of low-pressure ultraviolet light for inactivating Mycobacterium avium complex (MAC) micro-organisms

    Science.gov (United States)

    Aims: To assess low-pressure ultraviolet light (LP-UV) inactivation kinetics of Mycobacterium avium complex (MAC) strains in a water matrix using collimated beam apparatus. Methods and Results: Strains of M. avium (n = 3) and Mycobacterium intracellulare (n = 2) were exposed t...

  14. Characterization of the fibronectin-attachment protein of Mycobacterium avium reveals a fibronectin-binding motif conserved among mycobacteria.

    Science.gov (United States)

    Schorey, J S; Holsti, M A; Ratliff, T L; Allen, P M; Brown, E J

    1996-07-01

    Mycobacterium avium is an intracellular pathogen and a major opportunistic infectious agent observed in patients with acquired immune deficiency syndrome (AIDS). Evidence suggests that the initial portal of infection by M. avium is often the gastrointestinal tract. However, the mechanism by which the M. avium crosses the epithelial barrier is unclear. A possible mechanism is suggested by the ability of M. avium to bind fibronectin, an extracellular matrix protein that is a virulence factor for several extracellular pathogenic bacteria which bind to mucosal surfaces. To further characterize fibronectin binding by M. avium, we have cloned the M. avium fibronectin-attachment protein (FAP). The M. avium FAP (FAP-A) has an unusually large number of Pro and Ala residues (40% overall) and is 50% identical to FAP of both Mycobacterium leprae and Mycobacterium tuberculosis. Using recombinant FAP-A and FAP-A peptides, we show that two non-continuous regions in FAP-A bind fibronectin. Peptides from these regions and homologous sequences from M. leprae FAP inhibit fibronectin binding by both M. avium and Mycobacterium bovis Bacillus Calmette-Guerin (BCG). These regions have no homology to eukaryotic fibronectin-binding proteins and are only distantly related to fibronectin-binding peptides of Gram-positive bacteria. Nevertheless, these fibronectin-binding regions are highly conserved among the mycobacterial FAPs, suggesting an essential function for this interaction in mycobacteria infection of their metazoan hosts.

  15. KatG protein: A novel marker for differential diagnosis of Myobacterium avium complex infection

    Directory of Open Access Journals (Sweden)

    Gupta K

    2010-01-01

    Full Text Available Purpose: Biochemical or nucleic acid based diagnostic techniques for MAC infection are unsatisfactory. This study aims to identify and evaluate M. avium secretory protein(s of diagnostic potential, so as to develop a rapid and simple method for diagnosis of MAC infection. Material and Methods: Initially, a specific protein band of ~80-85 kDa was recognised by differential immunoblotting; which was subjected to anion exchange column chromatography for purification of proteins. After fractionisation using SDS-PAGE and electroelution, blast search was carried out. Further immunoreactivity studies were done with M. avium and Mtb infected mice sera. Clinical utilisation of separated protein was evaluated by conducting indirect ELISA with serum samples from mycobacterial infected patients. Results: A specific 81.6 kDa protein, shown to be catalase-peroxidase protein (KatG by blast search was separated. Immunoreactivity studies of purified KatG proteins with mice sera confirmed it to be specific for M. avium infection. Indirect ELISA with patient samples further confirmed it to be M. avium infection specific. Conclusion: KatG protein is specifically recognised by MAC patients and can be used as a marker for simple and rapid ELISA based tests for differential diagnosis of M. avium infection.

  16. Immunoreactivity of protein tyrosine phosphatase A (PtpA) in sera from sheep infected with Mycobacterium avium subspecies paratuberculosis.

    Science.gov (United States)

    Gurung, Ratna B; Begg, Douglas J; Purdie, Auriol C; Bach, Horacio; Whittington, Richard J

    2014-07-15

    Evasion of host defense mechanisms and survival inside infected host macrophages are features of pathogenic mycobacteria including Mycobacterium avium subspecies paratuberculosis, the causative agent of Johne's disease in ruminants. Protein tyrosine phosphatase A (PtpA) has been identified as a secreted protein critical for survival of mycobacteria within infected macrophages. The host may mount an immune response to such secreted proteins. In this study, the humoral immune response to purified recombinant M. avium subsp. paratuberculosis PtpA was investigated using sera from a cohort of sheep infected with M. avium subsp. paratuberculosis and compared with uninfected healthy controls. A significantly higher level of reactivity to PtpA was observed in sera collected from M. avium subspecies paratuberculosis infected sheep when compared to those from uninfected healthy controls. PtpA could be a potential candidate antigen for detection of humoral immune responses in sheep infected with M. avium subspecies paratuberculosis.

  17. Genome-wide analysis of the emerging infection with Mycobacterium avium subspecies paratuberculosis in the Arabian camels (Camelus dromedarius.

    Directory of Open Access Journals (Sweden)

    Pallab Ghosh

    Full Text Available Mycobacterium avium subspecies paratuberculosis (M. ap is the causative agent of paratuberculosis or Johne's disease (JD in herbivores with potential involvement in cases of Crohn's disease in humans. JD is spread worldwide and is economically important for both beef and dairy industries. Generally, pathogenic ovine strains (M. ap-S are mainly found in sheep while bovine strains (M. ap-C infect other ruminants (e.g. cattle, goat, deer, as well as sheep. In an effort to characterize this emerging infection in dromedary/Arabian camels, we successfully cultured M. ap from several samples collected from infected camels suffering from chronic, intermittent diarrhea suggestive of JD. Gene-based typing of isolates indicated that all isolates belong to sheep lineage of strains of M. ap (M. ap-S, suggesting a putative transmission from infected sheep herds. Screening sheep and goat herds associated with camels identified the circulation of this type in sheep but not goats. The current genome-wide analysis recognizes these camel isolates as a sub-lineage of the sheep strain with a significant number of single nucleotide polymorphisms (SNPs between sheep and camel isolates (∼1000 SNPs. Such polymorphism could represent geographical differences among isolates or host adaptation of M. ap during camel infection. To our knowledge, this is the first attempt to examine the genomic basis of this emerging infection in camels with implications on the evolution of this important pathogen. The sequenced genomes of M. ap isolates from camels will further assist our efforts to understand JD pathogenesis and the dynamic of disease transmission across animal species.

  18. 鸡肉空肠弯曲杆菌的分离鉴定及耐药性分析%Isolation, identification and antimicrobial resistance analysis of Campylobacter jejuni isolates originated from chicken

    Institute of Scientific and Technical Information of China (English)

    韩新锋; 刘书亮; 张晓利; 陈荀; 侯小刚

    2012-01-01

    目的 分离和收集食源性空肠弯曲杆菌的优势流行株,了解市场上鸡肉中空肠弯曲杆菌的污染和药物敏感性情况.方法 根据GB/T4789.9-2003、SN0175-2010、ISO10272-1∶2006、FDA/BAM∶2001、USDA/FSIS∶1998等方法优化组合出一套对动物性食品源空肠弯曲杆菌有效的分离鉴定方法,从市场上鸡肉中分离疑似空肠弯曲杆菌,并进一步用生化试验及PCR方法进行准确鉴定.采用微量肉汤稀释法测定了所分离的空肠弯曲杆菌对22种常见抗菌药物(组合)的最低抑菌浓度值(MIC).结果与结论 根据培养特征、形态特征、生化试验和PCR结果准确鉴定出21株空肠弯曲杆菌,其污染率为11.5%.86%生化试验阳性的菌株经PCR鉴定为阳性.将空肠弯曲杆菌标准菌株和随机选出的分离株的16S rDNA扩增序列进行比对,二者同源率为100%.药敏试验表明,14.3%的分离菌株对氨苄西林具有耐药性;大多数菌株对头孢类抗生素MIC值相对较高;42.9%的菌株对萘啶酸具有耐药性;所有分离株对环丙沙星、四环素、红霉素、氯霉素、庆大霉素、链霉素不耐药.%According to the standards of GB/T4789. 9-2003, SN0175-2010, ISO10272-1:2006, FDA/BAM : 2001 and USDA/FSIS : 1998, a series of effective isolation and identification methods was developed for Campylobacter jejuni of chicken origin. By using these methods, suspected Campylobacter jejuni was firstly isolated from chicken in the market of Ya'an City, Sichuan Province, and then biochemical test and PCR method were used to further confirm the isolation strains. Combining the results of cultural characteristics, morphologic characteristics, biochemical test and PCR results, 21 isolates were accurately identified as Campylobacter jejuni, with which the contamination incidence was 11. 5% (21/183). The 86% of the strains identified by biochemical test were further confirmed by PCR. The 16S rDNA sequence comparison of the

  19. Detecting local establishment strategies of wild cherry (Prunus avium L.

    Directory of Open Access Journals (Sweden)

    Gregorius Hans-Rolf

    2006-10-01

    Full Text Available Abstract Backround P. avium, a pioneer tree species that colonizes early forest successional stages, is assumed to require an effective strategy allowing stably repeatable rounds of local establishment, dispersal and local extinction. Consequently, the early replacement of cherry by climax tree species makes the establishment of several local generations very unlikely, especially in central European continuous cover forests. This has to be seen in connection with the mixed reproduction system involving asexual reproduction as a complementary adaptational strategy. Tests of the local establishment of wild cherry must therefore consider the possibility of first generation establishment via seedling recruitment potentially followed by an asexual generation (root suckering. Successful establishment can therefore be determined only among adult individuals with the option of detecting vegetative reproduction at these stages. To test the implied suggestion about local establishment strategies of wild cherry, nuclear microsatellites were used to analyse patterns of asexual propagation among adult stages that have been subjected to one of two major types of forest management. These management types, the historical "coppice with standards system" (CWS and the "high forest system" (HFS, can be reasonably assumed to have affected the reproduction system of P. avium. Results Clear differences were found in the reproduction pattern between two stands representing the two forest management types: 1 Clonal propagation is observed in both management systems, but with a distinctly higher frequency in the CWS. Hence, sexual recruitment as a first local generation is followed by a second asexual generation in both, whereas in the CWS there is evidence for an additional clonal generation. 2 The estimation of amounts of clonal reproduction critically depends on the assumptions about multilocus gene associations. This is revealed by the application of newly developed

  20. Over-expression of the PaAP1 gene from sweet cherry (Prunus avium L.) causes early flowering in Arabidopsis thaliana.

    Science.gov (United States)

    Wang, Jing; Zhang, Xiaoming; Yan, Guohua; Zhou, Yu; Zhang, Kaichun

    2013-02-15

    A homologue of SQUAMOSA/APETALA1, designated PaAP1, was isolated from Prunus avium by reverse transcription-PCR (RT-PCR). The full length of PaAP1 cDNA is 753 bp, and it codes for a polypeptide of 250 amino acid residues. Sequence comparison revealed that PaAP1 belongs to the MADS-box gene family. Phylogenetic analysis indicated that PaAP1 shared the highest identity with SQUA/AP1 homologues from Prunus serrulata. Real-time fluorescence quantitative PCR analysis showed that PaAP1 was expressed at high levels in petal, sepal, style, and flower buds, which was slightly different from the expression pattern of AP1 of Arabidopsis thaliana. To characterize the functions of PaAP1, we assessed Arabidopsis transformed with 35S::PaAP1. A total of 8 transgenic T(1) lines with an early flowering phenotype were obtained, and a 3:1 segregation ratio of flowering time was observed in the T(2) generation of 4 lines. This study provides the first functional analysis of an SQUA/AP1 homolog from P. avium and suggests that PaAP1 is potentially useful for shortening the juvenile period in sweet cherry.

  1. Evaluation of a bovine antibody test for diagnosing Mycobacterium avium complex in patients with cystic fibrosis

    DEFF Research Database (Denmark)

    Qvist, Tavs; Pressler, Tacjana; Katzenstein, Terese L;

    2017-01-01

    INTRODUCTION: The aim of this study was to test a commercial bovine enzyme-linked immunosorbent assay for investigating antibody activity against Mycobacterium avium complex. METHODS: All patients at the Copenhagen Cystic Fibrosis (CF) Center who had culture for nontuberculous mycobacteria...... before and after culture conversion was performed in case patients. RESULTS: Out of 286 included subjects, six had clinical M. avium complex pulmonary disease at the time of sera sampling. These patients presented with higher antibody test values (P-value ... at ruling out pulmonary disease. Screening sera from patients with CF could guide clinicians to focus attention on patients at higher risk of M. avium complex pulmonary disease. Pediatr Pulmonol. 2016; 9999:XX-XX. © 2016 Wiley Periodicals, Inc....

  2. Isolation of a monoclonal antibody that recognizes the origin binding domain of JCV, but not SV40, large T-antigen.

    Science.gov (United States)

    Grubman, Shelley A; Shin, Jong; Phelan, Paul J; Gong, Aaron; Can, Hande; Dilworth, Ryan; Kini, Sandeep Kuntadi; Gagnon, David; Archambault, Jacques; Meinke, Gretchen; Bohm, Andrew; Jefferson, Douglas M; Bullock, Peter A

    2016-10-01

    Within immunocompromised populations, the JC polyomavirus is the cause of the often-fatal disease Progressive Multifocal Leukoencephalopathy (PML). JC virus encodes a protein, termed T-antigen (T-ag), which is essential for its replication and pathogenicity. Previous studies of JCV T-ag have, in general, used antibodies raised against SV40 T-ag. Unfortunately, SV40 T-ag is also detected in humans and therefore there have been concerns about cross-reactivity. To address this issue, we have isolated a monoclonal antibody that binds to the JCV, but not the SV40, T-ag origin-binding domain (OBD). Furthermore, the region on the surface of the JCV T-ag OBD that is recognized by the "anti-JCV OBD mAb" has been mapped. We also demonstrate that the "anti-JCV OBD mAb" will be a useful reagent for standard techniques (e.g., Westerns blots and ELISAs). Finally, we note that additional monoclonal Abs that are specific for the T-ags encoded by the other human polyomaviruses could be generated by adopting the approach described herein.

  3. Genomic and phylogenetic analyses of an adenovirus isolated from a corn snake (Elaphe guttata) imply a common origin with members of the proposed new genus Atadenovirus.

    Science.gov (United States)

    Farkas, Szilvia L; Benko, Mária; Elo, Péter; Ursu, Krisztina; Dán, Adám; Ahne, Winfried; Harrach, Balázs

    2002-10-01

    Approximately 60% of the genome of an adenovirus isolated from a corn snake (Elaphe guttata) was cloned and sequenced. The results of homology searches showed that the genes of the corn snake adenovirus (SnAdV-1) were closest to their counterparts in members of the recently proposed new genus ATADENOVIRUS: In phylogenetic analyses of the complete hexon and protease genes, SnAdV-1 indeed clustered together with the atadenoviruses. The characteristic features in the genome organization of SnAdV-1 included the presence of a gene homologous to that for protein p32K, the lack of structural proteins V and IX and the absence of homologues of the E1A and E3 regions. These characteristics are in accordance with the genus-defining markers of atadenoviruses. Comparison of the cleavage sites of the viral protease in core protein pVII also confirmed SnAdV-1 as a candidate member of the genus ATADENOVIRUS: Thus, the hypothesis on the possible reptilian origin of atadenoviruses (Harrach, Acta Veterinaria Hungarica 48, 484-490, 2000) seems to be supported. However, the base composition of DNA sequence (>18 kb) determined from the SnAdV-1 genome showed an equilibrated GC content of 51%, which is unusual for an atadenovirus.

  4. Selection and characterisation of lactic acid bacteria isolated from different origins for ensilingRobinia pseudoacacia and Morus alba L. leaves

    Institute of Scientific and Technical Information of China (English)

    NI Kui-kui; YANG Hui-xiao; HUA Wei; WANG Yan-ping; PANG Hui-li

    2016-01-01

    The objective of this study was to isolate lactic acid bacteria (LAB) strains from different origins and to select the best strains for ensilingRobinia pseudoacacia (RB) and Morus albaL. (MB)leaves. The LAB strains were inoculated into the extracted liquid obtained from RB and MB leaves to evaluate the fermentation products. 11 LAB strains were selected for further experiments based on the highest products of lactic or acetic acid, including 1 strain of Weissela confusa, 2 ofLactobacilus reuteri and 8 ofLactobacilus plantarum.The API 50 CH fermentation experiment indicated that al of the selected 11 LAB strains utilised most of the carbohydrates. Al the strains grew at temperatures between 10 and 45°C and at a pH of 3.5 to 4.5; however,L. reuteri F7 and F8 tolerated a pH as low as 3.0. Al 11 LAB strains showed antibacterial activity against Listeria monocytogens,Escherichia coil,Salmonelasp. and Acetobacter pasteurianus; however, after excluding the effect of organic acids, only F7 and F8 stil exhibited antibacterial activity. The present study indicated that the selected 11 LAB strains could be used to prepare silages of RB and MB leaves, especialyL. reuteri F7 and F8.

  5. Volatile emissions from Mycobacterium avium subsp. paratuberculosis mirror bacterial growth and enable distinction of different strains.

    Directory of Open Access Journals (Sweden)

    Phillip Trefz

    Full Text Available Control of paratuberculosis in livestock is hampered by the low sensitivity of established direct and indirect diagnostic methods. Like other bacteria, Mycobacterium avium subsp. paratuberculosis (MAP emits volatile organic compounds (VOCs. Differences of VOC patterns in breath and feces of infected and not infected animals were described in first pilot experiments but detailed information on potential marker substances is missing. This study was intended to look for characteristic volatile substances in the headspace of cultures of different MAP strains and to find out how the emission of VOCs was affected by density of bacterial growth. One laboratory adapted and four field strains, three of MAP C-type and one MAP S-type were cultivated on Herrold's egg yolk medium in dilutions of 10(-0, 10(-2, 10(-4 and 10(-6. Volatile substances were pre-concentrated from the headspace over the MAP cultures by means of Solid Phase Micro Extraction (SPME, thermally desorbed from the SPME fibers and separated and identified by means of GC-MS. Out of the large number of compounds found in the headspace over MAP cultures, 34 volatile marker substances could be identified as potential biomarkers for growth and metabolic activity. All five MAP strains could clearly be distinguished from blank culture media by means of emission patterns based on these 34 substances. In addition, patterns of volatiles emitted by the reference strain were significantly different from the field strains. Headspace concentrations of 2-ethylfuran, 2-methylfuran, 3-methylfuran, 2-pentylfuran, ethyl acetate, 1-methyl-1-H-pyrrole and dimethyldisulfide varied with density of bacterial growth. Analysis of VOCs emitted from mycobacterial cultures can be used to identify bacterial growth and, in addition, to differentiate between different bacterial strains. VOC emission patterns may be used to approximate bacterial growth density. In a perspective volatile marker substances could be used to

  6. Crystal structure of sulfotransferase STF9 from Mycobacterium avium.

    Science.gov (United States)

    Hossain, Md Murad; Moriizumi, Yuuji; Tanaka, Shotaro; Kimura, Makoto; Kakuta, Yoshimitsu

    2012-02-01

    Sulfotransferases catalyze the sulfate conjugation of a wide variety of endogenous and exogenous molecules. Human pathogenic mycobacteria produce numerous sulfated molecules including sulfolipids which are well related to the virulence of several strains. The genome of Mycobacterium avium encodes eight putative sulfotransferases (stf1, stf4-stf10). Among them, STF9 shows higher similarity to human heparan sulfate 3-O-sulfotransferase isoforms than to the bacterial STs. Here, we determined the crystal structure of sulfotransferase STF9 in complex with a sulfate ion and palmitic acid at a resolution of 2.6 Å. STF9 has a spherical structure utilizing the classical sulfotransferase fold. STF9 exclusively possesses three N-terminal α-helices (α1, α2, α3) parallel to the 3'-phosphoadenosine-5'-phosphosulfate (PAPS) binding motif. The sulfate ion binds to the PAPS binding structural motif and the palmitic acid molecule binds in the deep cleft of the predicted substrate binding site suggesting the nature of endogenous acceptor substrate of STF9 resembles palmitic acid. The substrate binding site is covered by a flexible loop which may have involvement in endogenous substrate recognition. Based on the mutational study (Hossain et al., Mol Cell Biochem 350:155-162; 2011) and structural resemblance of STF9-sulfate ion-palmitic acid complex to the hHS3OST3 complex with PAP (3'-phosphoadenosine-5'-phosphate) and an acceptor sugar chain, Glu170 and Arg96 are appeared to be catalytic residues in STF9 sulfuryl transfer mechanism.

  7. Ascorbic acid metabolism during sweet cherry (Prunus avium) fruit development.

    Science.gov (United States)

    Liang, Dong; Zhu, Tingting; Ni, Zhiyou; Lin, Lijin; Tang, Yi; Wang, Zhihui; Wang, Xun; Wang, Jin; Lv, Xiulan; Xia, Hui

    2017-01-01

    To elucidate metabolism of ascorbic acid (AsA) in sweet cherry fruit (Prunus avium 'Hongdeng'), we quantified AsA concentration, cloned sequences involved in AsA metabolism and investigated their mRNA expression levels, and determined the activity levels of selected enzymes during fruit development and maturation. We found that AsA concentration was highest at the petal-fall period (0 days after anthesis) and decreased progressively during ripening, but with a slight increase at maturity. AsA did nevertheless continue to accumulate over time because of the increase in fruit fresh weight. Full-length cDNAs of 10 genes involved in the L-galactose pathway of AsA biosynthesis and 10 involved in recycling were obtained. Gene expression patterns of GDP-L-galactose phosphorylase (GGP2), L-galactono-1, 4-lactone dehydrogenase (GalLDH), ascorbate peroxidase (APX3), ascorbate oxidase (AO2), glutathione reductase (GR1), and dehydroascorbate reductase (DHAR1) were in accordance with the AsA concentration pattern during fruit development, indicating that genes involved in ascorbic acid biosynthesis, degradation, and recycling worked in concert to regulate ascorbic acid accumulation in sweet cherry fruit.

  8. Control of Mycobacterium avium subsp. paratuberculosis infection in agricultural species.

    Science.gov (United States)

    Kennedy, D J; Benedictus, G

    2001-04-01

    Paratuberculosis or Johne's disease is a chronic intestinal disease caused by Mycobacterium avium subsp. paratuberculosis, which continues to spread in agricultural species. Control of paratuberculosis is challenging and should not be underestimated. Due to the long incubation period of the infection, disease is largely subclinical in domesticated livestock. Hence, direct effects on animal productivity and welfare are often masked and may appear insufficient to justify large investments in control programmes by individual farmers, livestock industries or governments. Furthermore, in some countries the main effects of the disease are indirect, resulting from the impact of market discrimination against herds and flocks known to be infected, or from the control measures enforced to reduce transmission. In such circumstances, producers may be unwilling to co-operate with surveillance that may detect infection in herds or flocks. As control programmes are rarely successful in eliminating the infection from a herd or flock in the short term without an aggressive and costly programme, financial and community support assists producers to deal with the challenge. Successful prevention and control depends on animal health authorities and livestock industries acquiring a good understanding of the nature and epidemiology of infection, and of the application of tools for diagnosis and control. Building support for control programmes under the leadership of the affected livestock industries is critical, as programmes are unlikely to be successful without ongoing political will, supported by funding for research, surveillance and control.

  9. Description of a Novel Adhesin of Mycobacterium avium Subsp. paratuberculosis

    Directory of Open Access Journals (Sweden)

    Mariana Noelia Viale

    2014-01-01

    Full Text Available The binding and ingestion of Mycobacterium avium subsp. paratuberculosis (MAP by host cells are fibronectin (FN dependent. In several species of mycobacteria, a specific family of proteins allows the attachment and internalization of these bacteria by epithelial cells through interaction with FN. Thus, the identification of adhesion molecules is essential to understand the pathogenesis of MAP. The aim of this study was to identify and characterize FN binding cell wall proteins of MAP. We searched for conserved adhesins within a large panel of surface immunogenic proteins of MAP and investigated a possible interaction with FN. For this purpose, a cell wall protein fraction was obtained and resolved by 2D electrophoresis. The immunoreactive spots were identified by MALDI-TOF MS and a homology search was performed. We selected elongation factor Tu (EF-Tu as candidate for further studies. We demonstrated the FN-binding capability of EF-Tu using a ligand blot assay and also confirmed the interaction with FN in a dose-dependent manner by ELISA. The dissociation constant of EF-Tu was determined by surface plasmon resonance and displayed values within the μM range. These data support the hypothesis that this protein could be involved in the interaction of MAP with epithelial cells through FN binding.

  10. Complete Remission of Minimal Change Disease Following an Improvement of Lung Mycobacterium avium Infection.

    Science.gov (United States)

    Yamashiro, Aoi; Uchida, Takahiro; Ito, Seigo; Oshima, Naoki; Oda, Takashi; Kumagai, Hiroo

    A 46-year-old woman suddenly developed peripheral edema. Her massive proteinuria, hypoproteinemia, and renal biopsy findings yielded the diagnosis of minimal change disease (MCD). In addition, lung Mycobacterium avium infection was diagnosed according to a positive culture of her bronchoalveolar lavage fluid. The lung lesion was improved by anti-nontuberculous mycobacteria therapy. Surprisingly, her proteinuria also gradually decreased and she attained complete remission of MCD without any immunosuppressive therapy. She has subsequently remained in complete remission. We herein report an interesting case of MCD with lung Mycobacterium avium infection, suggesting a causal relationship among infection, immune system abnormality, and MCD/nephrotic syndrome.

  11. A novel multi-antigen virally vectored vaccine against Mycobacterium avium subspecies paratuberculosis.

    Directory of Open Access Journals (Sweden)

    Tim J Bull

    Full Text Available BACKGROUND: Mycobacterium avium subspecies paratuberculosis causes systemic infection and chronic intestinal inflammation in many species including primates. Humans are exposed through milk and from sources of environmental contamination. Hitherto, the only vaccines available against Mycobacterium avium subspecies paratuberculosis have been limited to veterinary use and comprised attenuated or killed organisms. METHODS: We developed a vaccine comprising a fusion construct designated HAV, containing components of two secreted and two cell surface Mycobacterium avium subspecies paratuberculosis proteins. HAV was transformed into DNA, human Adenovirus 5 (Ad5 and Modified Vaccinia Ankara (MVA delivery vectors. Full length expression of the predicted 95 kDa fusion protein was confirmed. PRINCIPAL FINDINGS: Vaccination of naïve and Mycobacterium avium subspecies paratuberculosis infected C57BL/6 mice using DNA-prime/MVA-boost or Ad5-prime/MVA-boost protocols was highly immunogenic resulting in significant IFN-gamma ELISPOT responses by splenocytes against recombinant vaccine antigens and a range of HAV specific peptides. This included strong recognition of a T-cell epitope GFAEINPIA located near the C-terminus of the fusion protein. Antibody responses to recombinant vaccine antigens and HAV specific peptides but not GFAEINPIA, also occurred. No immune recognition of vaccine antigens occurred in any sham vaccinated Mycobacterium avium subspecies paratuberculosis infected mice. Vaccination using either protocol significantly attenuated pre-existing Mycobacterium avium subspecies paratuberculosis infection measured by qPCR in spleen and liver and the Ad5-prime/MVA-boost protocol also conferred some protection against subsequent challenge. No adverse effects of vaccination occurred in any of the mice. CONCLUSIONS/SIGNIFICANCE: A range of modern veterinary and clinical vaccines for the treatment and prevention of disease caused by Mycobacterium avium

  12. PD-L2 induction on dendritic cells exposed to Mycobacterium avium downregulates BCG-specific T cell response.

    Science.gov (United States)

    Mendoza-Coronel, Elizabeth; Camacho-Sandoval, Rosa; Bonifaz, Laura C; López-Vidal, Yolanda

    2011-01-01

    The exposure to certain species of Nontuberculous Mycobacteria (NTM) can modulate the immune response induced by Mycobacterium bovis BCG. Mycobacterium avium has been postulated as a weak inducer of dendritic cell (DC) maturation. However, how the DC exposure to M. avium could contribute to the modulation of a BCG-specific CD4+ T cell response and the molecules involved remain unknown. Here, we exposed bone marrow-derived DCs (BMDCs) to M. avium either prior to exposure to BCG or as a unique stimulus. We found that M. avium induces high expression of PD-L2 (B7-DC) in BMDCs. This was dependent on IL-10 production through the TLR2-p38 MAPK signaling pathway. Exposure to M. avium prior to BCG results in BMDCs that do not express co-stimulatory molecules and pro-inflammatory cytokines, while the expression of PD-L2 and IL-10 was maintained. BMDCs exposed to M. avium impaired the activation of BCG-specific T cells through the PD-1: PD-L interaction. This suggests that a M. avium-induced phenotype in DCs might be implicated in the induction of mechanisms of tolerance that could impact the T cell response induced by BCG vaccination.

  13. Bioactive components of Prunus avium L. black gold (red cherry) and Prunus avium L. stark gold (white cherry) juices, wines and vinegars.

    Science.gov (United States)

    Budak, Nilgün H

    2017-01-01

    Cherries are one of the most popular fruits, characterized by attractive colour, firmness, appearance and delicious tastes. Cherries are consumed fresh as well as in jams, wine, dried, candy and other processed products. Cherries vary in antioxidant properties and phenolic substances. The aim of the study was to determine the effects of ethanol and acetic acid fermentation on total antioxidant activities and phenolic substances of cherry juice. Total investigation of solids, pH, soluble solids, phenolic substances, ORAC and TEAC of Prunus avium L. cherry juices, macerated cherries wine, and vinegars were analyzed. All samples had 300.1-854.79 mg GAE/L of total phenolic contents, and 6.62-17.97 µmol/mL of ORAC values, and 1.5-5.5 mmol/mL of TEAC. Chlorogenic acid was present in the highest amount P. avium L. black gold vinegar.

  14. [Isolation and Identification of a Quail-origin H9N2 Subtype of The Influenza Virus and Its Biologic Characterization].

    Science.gov (United States)

    Yu, Yang; Si, Weiying; Yuan, Zhuangchuan; Yan, Yan; Zhou, Jiyong

    2016-01-01

    A quail-origin subtype of the influenza virus was isolated from a human-infecting H7N9 subtype of the avian influenza virus found in a live poultry market and was given the name A/Quail/Hangzhou/1/ 2013 (H9N2). We analyzed the whole genome of this virus and its biologic characteristics. Sequence analyses suggested that the: HA and NS genes belonged to a CK/BJ/1/94-like lineage; NA, NP, PA and PB1 genes belonged to a SH/F/98-like lineage; M and PB2 genes belonged to a G1-like lineage. Analyses of key amino acids showed that the cleavage site in HA protein was PSRSSR ↓ GL, and that the HA protein had a human receptor-binding site with Leu226. Deletion of amino acids 69 - 73 was detected in the stalk of NA protein, the M2 protein had an Asn31 mutation, and the NS1 protein had two mutations at Ser42, Ala149. The intravenous pathogenicity of this virus was 0.36. A study in chickens suggested that all inoculated birds shed the virus from the trachea and cloaca on the third day post-infection (p. i. ) until 11 days. All chickens that had direct contact shed the virus on the second day p. i. until 8 days. Results of virus reisolation suggested that lung and tracheal tissues could shed the virus in 5 days, whereas the other organs could shed the virus in 3 days. These results suggest that this virus strain is H9N2 subtype LPAIV, whose lineage is prevalent in mainland China. This research provides evidence on how to monitor and prevent the H9N2 subtype of the avian influenza virus.

  15. Examination of Mycobacterium avium subspecies paratuberculosis mixed genotype infections in dairy animals using a whole genome sequencing approach.

    Science.gov (United States)

    Davidson, Fraser W; Ahlstrom, Christina; De Buck, Jeroen; Whitney, Hugh G; Tahlan, Kapil

    2016-01-01

    Many pathogenic mycobacteria are known to cause severe disease in humans and animals. M. avium subspecies paratuberculosis (Map) is the causative agent of Johne's disease-a chronic wasting disease affecting ruminants such as cattle and sheep, responsible for significant economic losses in the dairy and beef industries. Due to the lack of treatment options or effective vaccines, mitigating losses can be difficult. In addition, the early stages of Map infection may occur in asymptomatic hosts that continue to shed viable bacteria in their faeces, leading to the infection of other healthy animals. Using multi-locus short sequence repeat (ML-SSR) analysis we previously reported that individual Johne's positive dairy cattle from farms across the island of Newfoundland were infected by Map with multiple SSR-types simultaneously. The occurrence of multiple mixed genotype infections has the potential to change pathogen and disease dynamics as well as reduce the efficacy of treatments and vaccines. Therefore, we conducted whole genome sequencing (WGS) and single nucleotide polymorphism (SNP) analysis on a subset of these isolates for a more in-depth examination. We also implemented a PCR assay using two discriminatory SNPs and demonstrated the incidence of a mixed infection by three genotypically diverse Map isolates in a single animal. In addition, results show that WGS and SNP analysis can provide a better understanding of the relationship between Map isolates from individual and different animals. In the future such studies on the occurrence of mixed genotype infections could potentially lead to the identification of variable pathogenicity of different genotypes and allow for better tracking of Map isolates for epidemiological studies.

  16. Mycobacterium avium-intracellulare infection during HIV disease. Persisting problems

    Directory of Open Access Journals (Sweden)

    Roberto Manfredi

    2008-12-01

    Full Text Available Still in the era of combined antiretroviral therapy, late recognition of HIV disease or lack of sufficient immune recovery pose HIV-infected patients at risk to develop opportunistic infections by nontuberculous mycobacteria (NTM, which are environmental organisms commonly retrieved in soil and superficial waters.Among these microorganisms, the most frequent is represented by Mycobacterium avium complex (MAC. Health care professionals who face HIV-infected patients should suspect disseminated mycobacterial disease when a deep immunodeficiency is present, (a CD4+ lymphocyte count below 50 cells/μL often associated with constitutional signs and symptoms, and non-specific laboratory abnormalities. Mycobacterial culture of peripheral blood is a reliable technique for diagnosing disseminated disease. Among drugs active against NTM, as well as some anti-tubercular compounds, the rifampin derivative rifabutin, and some novel fluoroquinolones, the availability of macrolides, has greatly contributed to improve both prophylaxis and treatment outcome of disseminated MAC infections. Although multiple questions remain about which regimens may be regarded as optimal, general recommendations can be expressed on the ground of existing evidences.Treatment should begin with associated clarithromycin (or azithromycin, plus ethambutol and rifabutin (with the rifabutin dose depending on other concomitant medications that might result in drug-drug interactions.A combined three-drug regimen is preferred for patients who cannot be prescribed an effective antiretroviral regimen immediately. Patients with a CD4+ lymphocyte count below 50 cells/μL, who do not have clinical evidence of active mycobacterial disease, should receive a primary prophylaxis with either clarithromycin or azithromycin, with or without rifabutin.

  17. Interaction between Mycobacterium avium subsp. paratuberculosis and environmental protozoa

    Directory of Open Access Journals (Sweden)

    Rowe Michael T

    2006-07-01

    Full Text Available Abstract Background Interactions between Mycobacterium avium subsp. paratuberculosis (Map and free-living protozoa in water are likely to occur in nature. The potential impact of ingestion of Map by two naturally occurring Acanthamoeba spp. on this pathogen's survival and chlorine resistance was investigated. Results Between 4.6 and 9.1% of spiked populations of three Map strains (NCTC 8578, B2 and ATCC 19698, which had been added at a multiplicity of infection of 10:1, were ingested by Acanthamoeba castellanii CCAP 1501/1B and A. polyphaga CCAP 1501/3B during co-culture for 3 h at 25°C. Map cells were observed to be present within the vacuoles of the amoebae by acid-fast staining. During extended co-culture of Map NCTC 8578 at 25°C for 24 d with both A. castellanii and A. polyphaga Map numbers did not change significantly during the first 7 days of incubation, however a 1–1.5 log10 increase in Map numbers was observed between days 7 and 24 within both Acanthamoeba spp. Ingested Map cells were shown to be more resistant to chlorine inactivation than free Map. Exposure to 2 μg/ml chlorine for 30 min resulted in a log10 reduction of 0.94 in ingested Map but a log10 reduction of 1.73 in free Map (p Conclusion This study demonstrated that ingestion of Map by and survival and multiplication of Map within Acanthamoeba spp. is possible, and that Map cells ingested by amoebae are more resistant to inactivation by chlorine than free Map cells. These findings have implications with respect to the efficacy of chlorination applied to Map infected surface waters.

  18. The relative frequency of Mycobacterium tuberculosis and Mycobacterium avium infections in HIV positive patients, Ahvaz, Iran

    Institute of Scientific and Technical Information of China (English)

    Khosravi AD; Alavi SM; Hashemzade M; Abasi E; Seghatoleslami S

    2012-01-01

    Objective:To estimate the prevalence of Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium avium (M. avium) infections in HIV-positive patients suspected to have pulmonary and extrapulmonary mycobacterial co-infection using PCR technique. Methods:Totally 50 samples comprising sputum, pleural fluid and CSF taken from HIV positive patients suspected to have mycobacterial infection, were processed. The demographic information and results of acid fast staining and culture were recorded for each patient. The PCR for detecting of M. tuberculosis comprised of specific primers targeting IS6110 gene sequence. For detecting of M. avium, PCR with primers that amplifies the mig gene were used. Results:From 50 samples processed, 45 were sputum (90%), 3 pleural fluid (6%) and 2 CSF (4%). In total, 8 (16%) were culture positive, 7 had positive acid fast staining (14%) and 13 samples (26%) were positive using PCR technique. All the positive samples were sputum and belonged to patients with pulmonary infection. Of these, 9 were positive for M. tuberculosis (69.2%) and 4 were identified as M. avium (30.8%), which 2 out of 13 positive samples showed mixed infections by both mycobacteria. Conclusions:The PCR shows the highest detection rate (26%) of mycobacteria compared with culture and acid fast staining. The majority of infections were with M. tuberculosis (18%) and this shows the importance of this mycobacterial co-infection in HIV positive patients in the region of study.

  19. Bordetella avium causes induction of apoptosis and nitric oxide synthase in turkey tracheal explant cultures.

    Science.gov (United States)

    Miyamoto, David M; Ruff, Kristin; Beach, Nathan M; Stockwell, Stephanie B; Dorsey-Oresto, Angella; Masters, Isaac; Temple, Louise M

    2011-09-01

    Bordetellosis is an upper respiratory disease of turkeys caused by Bordetella avium in which the bacteria attach specifically to ciliated respiratory epithelial cells. Little is known about the mechanisms of pathogenesis of this disease, which has a negative impact in the commercial turkey industry. In this study, we produced a novel explant organ culture system that was able to successfully reproduce pathogenesis of B. avium in vitro, using tracheal tissue derived from 26 day-old turkey embryos. Treatment of the explants with whole cells of B. avium virulent strain 197N and culture supernatant, but not lipopolysaccharide (LPS) or tracheal cytotoxin (TCT), specifically induced apoptosis in ciliated cells, as shown by annexin V and TUNEL staining. LPS and TCT are known virulence factors of Bordetella pertussis, the causative agent of whooping cough. Treatment with whole cells of B. avium and LPS specifically induced NO response in ciliated cells, shown by uNOS staining and diaphorase activity. The explant system is being used as a model to elucidate specific molecules responsible for the symptoms of bordetellosis.

  20. Kirsipuu (Prunus avium) : [luuletused] / R. W. Stedingh ; tlk. ja saatesõna: Jüri Talvet

    Index Scriptorium Estoniae

    Stedingh, R. W.

    2003-01-01

    Sisu: Kirsipuu (Prunus avium) ; Rubus spectabilis ; Rododendron (Rhododendron macrophyllum) ; Lysuchitum americanum ; Tulp (Tulipa gesneriana) ; Kanada hani (Branta canadensis) ; Metsorava pärastlõuna (Sciurus carolinensis) ; Ohakalind (Spinus tristis) ; Shakespeare'i mälestusmärk (kogust "Stanley pargi süit")

  1. Sensitive detection of Myobacterium avium subsp paratuberculosis in bovine semen by real-time PCR

    NARCIS (Netherlands)

    Herthnek, D.; Englund, S.; Willemsen, P.T.J.; Bolske, G.

    2006-01-01

    Aims: To develop a fast and sensitive protocol for detection of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine semen and to make a critical evaluation of the analytical sensitivity. Methods and Results: Processed semen was spiked with known amounts of MAP. Semen from different bulls as

  2. THE EFFECT OF TEMPERATURE ON THE GROWTH OF MYCOBACTERIUM AVIUM COMPLEX (MAC) ORGANISMS

    Science.gov (United States)

    MAC organisms are able to grow, persist, and colonize in water distribution systems and may amplify in hospital hot water systems. This study examined the response of MAC organisms (M. avium, M. intracellulare, and MX) to a range of temperatures commonly associated with drinking...

  3. Detection of Mycobacterium avium subspecies paratuberculosis in Drinking Water and Biofilms Using Quantitative PCR

    Science.gov (United States)

    Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne’s disease in domestic animals and has been implicated in Crohn’s disease in humans. This bacterium is a slow growing, gram-positive, acid-fast organism which can be difficult to culture from the environment. For ...

  4. IL-32 expression in the airway epithelial cells of patients with Mycobacterium avium complex lung disease.

    NARCIS (Netherlands)

    Bai, X.; Ovrutsky, A.R.; Kartalija, M.; Chmura, K.; Kamali, A.; Honda, J.R.; Oberley-Deegan, R.E.; Dinarello, C.A.; Crapo, J.D.; Chang, L.Y.; Chan, E.D.

    2011-01-01

    Lung disease due to Mycobacterium avium complex (MAC) organisms is increasing. A greater understanding of the host immune response to MAC organisms will provide a foundation to develop novel therapies for these recalcitrant infections. IL-32 is a newly described pro-inflammatory cytokine that enhanc

  5. Detection of Mycobacterium avium subsp. paratuberculosis in milk from clinically affected cows by PCR and culture

    DEFF Research Database (Denmark)

    Giese, Steen Bjørck; Ahrens, Peter

    2000-01-01

    Milk and faeces samples from cows with clinical symptoms of paratuberculosis were examined for the presence of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) by culture and PCR. M. paratuberculosis was cultivated in variable numbers from faeces or intestinal mucosa in eight of 11...

  6. Genetic analysis of Mycobacterium avium complex strains used for producing purified protein derivatives

    NARCIS (Netherlands)

    Semret, M.; Bakker, D.; Smart, N.; Olsen, I.; Haslov, K.; Behr, M.A.

    2006-01-01

    For over a century, purified protein derivatives (PPD) have been used to detect mycobacterial infections in humans and livestock. Among these, reagents to detect infections by Mycobacterium avium complex organisms have been produced, but the utility of these reagents has not been clearly established

  7. Antibodies Induced by Lipoarabinomannan in Bovines: Characterization and Effects on the Interaction between Mycobacterium avium Subsp. paratuberculosis and Macrophages In Vitro

    Directory of Open Access Journals (Sweden)

    Ana Jolly

    2011-01-01

    Full Text Available Lipoarabinomannan (LAM is a major glycolipidic antigen on the mycobacterial envelope. The aim of this study was to characterize the humoral immune response induced by immunization with a LAM extract in bovines and to evaluate the role of the generated antibodies in the in vitro infection of macrophages with Mycobacterium avium subsp. paratuberculosis (MAP. Sera from fourteen calves immunized with LAM extract or PBS emulsified in Freund's Incomplete Adjuvant and from five paratuberculosis-infected bovines were studied. LAM-immunized calves developed specific antibodies with IgG1 as the predominant isotype. Serum immunoglobulins were isolated and their effect was examined in MAP ingestion and viability assays using a bovine macrophage cell line. Our results show that the antibodies generated by LAM immunization significantly increase MAP ingestion and reduce its intracellular viability, suggesting an active role in this model.

  8. Antibodies Induced by Lipoarabinomannan in Bovines: Characterization and Effects on the Interaction between Mycobacterium Avium Subsp. Paratuberculosis and Macrophages In Vitro

    Science.gov (United States)

    Jolly, Ana; Colavecchia, Silvia Beatriz; Fernández, Bárbara; Fernández, Eloy; Mundo, Silvia Leonor

    2011-01-01

    Lipoarabinomannan (LAM) is a major glycolipidic antigen on the mycobacterial envelope. The aim of this study was to characterize the humoral immune response induced by immunization with a LAM extract in bovines and to evaluate the role of the generated antibodies in the in vitro infection of macrophages with Mycobacterium avium subsp. paratuberculosis (MAP). Sera from fourteen calves immunized with LAM extract or PBS emulsified in Freund's Incomplete Adjuvant and from five paratuberculosis-infected bovines were studied. LAM-immunized calves developed specific antibodies with IgG1 as the predominant isotype. Serum immunoglobulins were isolated and their effect was examined in MAP ingestion and viability assays using a bovine macrophage cell line. Our results show that the antibodies generated by LAM immunization significantly increase MAP ingestion and reduce its intracellular viability, suggesting an active role in this model. PMID:21772964

  9. PARATUBERCULOSIS AND FOOD OF ANIMAL ORIGIN

    Directory of Open Access Journals (Sweden)

    T.A. Sarli

    2013-02-01

    Full Text Available Mycobacterium avium ssp. paratubercolosis (MAP is the causative agent of Johne’s disease (or paratubercolosis,a chronic infectious enteritis in cattle, sheep and goats. Infected animals shed viable MAP in their milk, faeces and semen. MAP may have a role in development of Crohn’s disease,a chronic inflammatory bowel disease in humans, via the consumption of contamined milk and milk products, meat and contamined water supplies. For some authors pasteurization is not sufficient to kill all MAP cells present in milk and it has been cultured from raw or pasteurizated milk and isolated from cheese. MAP has not isolated from retail beef to date, although limited testing has been carried out. Probably MAP may be involved in other chronic diseases like Type-1 Diabetes. Which is the possible public health consequence of periodically use by susceptible individuals is uncertain.

  10. Mycobacterium avium subsp. paratuberculosis survival during fermentation of soured milk products detected by culture and quantitative real time PCR methods.

    Science.gov (United States)

    Klanicova, B; Slana, I; Roubal, P; Pavlik, I; Kralik, P

    2012-07-02

    Mycobacterium avium paratuberculosis (MAP), etiological agent of paratuberculosis in ruminants, is able to survive extreme conditions like very low pH (stomach), high temperature (pasteurization) or low temperature (refrigerated storage). Cheese, infant powder milk, cream and other milk and dairy products might thus be considered as possible sources of MAP for humans. The aim of this study was to investigate the survival of two MAP field isolates during fermentation of three different types of soured milk products (SMP; yogurt, acidophilus milk and kefir) under laboratory conditions. Pasteurized MAP-free milk was artificially contaminated with 10(6)MAPcells/mL and survival and absolute numbers of MAP were monitored during fermentation (4 or 16 h) and after six weeks of storage at 4°C by culture and quantitative real time PCR (qPCR). Viability of MAP was determined by culture using Herrold's egg yolk medium and Middlebrook 7H10 with antibiotics, supplemented with Mycobactin J and incubated at 37°C for up to 12 weeks. The absolute numbers of MAP were quantified by previously published qPCR assays targeting F57 and IS900 loci in MAP genome. We herein confirm that MAP can survive pH reduction, however, longer exposure to pH below 4 in SMP seems to be critical because it inhibits growth. Therefore, it is suggested that probiotic cultures that can decrease pH below 4 during fermentation could provide better inactivation of MAP in SMP.

  11. Effects of nutritional and ambient oxygen condition on biofilm formation in Mycobacterium avium subsp. hominissuis via altered glycolipid expression

    Science.gov (United States)

    Totani, Takahiro; Nishiuchi, Yukiko; Tateishi, Yoshitaka; Yoshida, Yutaka; Kitanaka, Hiromi; Niki, Mamiko; Kaneko, Yukihiro; Matsumoto, Sohkichi

    2017-01-01

    Mycobacterium avium subsp. hominissuis (MAH) is the major causative agent of nontuberculous mycobacteriosis, the representative case of environment-related infectious diseases the incidence of which is increasing in industrialized countries. MAH is found in biofilm in drinking water distribution system and residential environments. We investigated the effect of gaseous and nutritional conditions, and the role of glycopeptidolipids (GPLs) on biofilm-like pellicle formation in MAH. Pellicle formation was observed under 5% oxygen in Middlebrook 7H9 broth containing 0.2% glycerol and 10% albumin-dextrose-catalase enrichment but not under normoxia or in nutrient-poor media. An analysis of 17 environmental isolates revealed that hypoxia (5% oxygen) preferentially enhanced pellicle formation both in plastic plates and in glass tubes, compared with hypercapnia (5% carbon dioxide). Wild-type strains (WT) developed much thicker pellicles than GPL-deficient rough mutants (RM). WT bacterial cells distributed randomly and individually in contrast to that RM cells positioned linearly in a definite order. Exogenous supplementation of GPLs thickened the pellicles of RM, resulting in a similar morphological pattern to WT. These data suggest a significant implication of eutrophication and hypoxia in biofilm-like pellicle formation, and a functional role of GPLs on development of pellicles in MAH. PMID:28155911

  12. In vitro cytokine profiles and viability of different human cells treated with whole cell lysate of Mycobacterium avium subsp. paratuberculosis

    Directory of Open Access Journals (Sweden)

    Rani Pittu

    2012-09-01

    Full Text Available Abstract Mycobacterium avium subsp. paratuberculosis (MAP is a zoonotic pathogen, a very slow growing bacterium which is difficult to isolate and passage in conventional laboratory culture. Although its association with Johne’s disease or paratuberculosis of cattle is well established, it has been only putatively linked to Crohn’s disease in humans. Further, MAP has been recently suggested to be a trigger for other autoimmune diseases such as type-1 diabetes mellitus (T1DM. Recently, some studies have indicated that exposure to MAP is associated with elevated levels of antibodies against MAP lysate although the exact mechanism and significance of the same remains unclear. Further, the cytokine profiles relevant in MAP associated diseases of humans and their exact role in the pathophysiology are not clearly known. We performed in vitro cytokine analyses after exposing different cultured human cells to the whole cell lysate of MAP and found that MAP lysate induces secretion of cytokines IL-1β, IL-6, IL-8, IL-10 and TNF-α by human peripheral blood mononuclear cells (PBMCs. Also, it induces secretion of IL-8 by cultured human stomach adenocarcinoma cells (AGS and PANC-1(human pancreatic carcinoma cell line cells. We also found that MAP lysate induced cytotoxicity in PANC-1cells. Collectively, these results provide a much needed base-line data set of cytokines broadly signifying a MAP induced cellular response by human cells.

  13. A Comparative Study of Detection of Bordetella avium Antibodies in Turkeys by ELISA, SPAT, and AGID Test

    OpenAIRE

    TÜRKYILMAZ, Süheyla; TÜRKYILMAZ, Kenan; KAYA, Osman

    2006-01-01

    The aims of this study were to develop a serum plate agglutination test (SPAT) antigen and agar gel immunodiffusion (AGID) test antigen for the serological detection of turkeys that have been exposed to Bordetella avium; to compare the sensitivity of commercial enzyme-linked immunosorbent assay (ELISA) with SPAT, and AGID test, and to survey B. avium antibodies in turkey flocks in Aydın, Turkey. For these purposes, serum samples collected from 300 turkeys were examined by ELISA, SPAT, and AGI...

  14. Palatal Actinomycosis and Kaposi Sarcoma in an HIV-Infected Subject with Disseminated Mycobacterium avium-intracellulare Infection

    Directory of Open Access Journals (Sweden)

    Yuria Ablanedo-Terrazas

    2012-01-01

    Full Text Available Actinomyces and Mycobacterium avium-intracellulare are facultative intracellular organisms, members of the bacterial order actinomycetales. Although Actinomyces can behave as copathogen when anatomic barriers are compromised, its coinfection with Mycobacterium avium-intracellulare has not previously been reported. We present the first reported case of palatal actinomycosis co-infection with disseminated MAC, in an HIV-infected subject with Kaposi sarcoma and diabetes. We discuss the pathogenesis of the complex condition of this subject.

  15. Evaluation of the MycoAKT latex agglutination test for rapid diagnosis of Mycobacterium avium complex infections.

    Science.gov (United States)

    Olano, J P; Holmes, H; Woods, G L

    1998-01-01

    Rapid diagnosis of Mycobacterium avium complex (MAC) bacteremia is important for management of patients with the acquired immunodeficiency syndrome who have disseminated MAC. The purpose of this study was to determine the reliability of the MycoAKT latex agglutination test for direct detection of MAC in positive mycobacterial blood cultures. First, colonies of isolates of previously identified mycobacteria, including 35 MAC, were tested. Of the 55 isolates evaluated, 33 were identified as MAC by the latex test, including 31 of the known MAC and 2 M. chelonae (sensitivity, 88.6%; specificity, 90.0%). Second, broth from 20 ESP II and 20 BACTEC 12B bottles seeded with isolates of MAC were tested. Aliquots from 19 (95%) ESP II cultures and 16 (80%) 12B cultures were positive by the latex test. In phase 3, broth from 115 signal-positive ESP II blood cultures were tested by latex agglutination. Forty-three subcultures from these bottles grew mycobacteria (41 MAC and 2 Mycobacterium tuberculosis complex); the remainder grew no organisms. Broth from 40 of the blood cultures (39 that grew MAC and 1 from which no organisms were recovered) were latex positive; thus, the sensitivity, specificity, and positive and negative predictive values of the latex test for direct identification of MAC in ESP II blood cultures were 95.1, 98.6, 97.5, and 97.3%, respectively. The mean time to detection of MAC was 14.6 days (range, 6-34 days) with the direct latex test, compared with 18.3 days (range, 9-36 days) with subculture and probe (p < 0.05).

  16. Efficacy of Various Pasteurization Time-Temperature Conditions in Combination with Homogenization on Inactivation of Mycobacterium avium subsp. paratuberculosis in Milk

    OpenAIRE

    Grant, Irene R.; Williams, Alan G.; Rowe, Michael T.; Muir, D. Donald

    2005-01-01

    The effect of various pasteurization time-temperature conditions with and without homogenization on the viability of Mycobacterium avium subsp. paratuberculosis was investigated using a pilot-scale commercial high-temperature, short-time (HTST) pasteurizer and raw milk spiked with 101 to 105 M. avium subsp. paratuberculosis cells/ml. Viable M. avium subsp. paratuberculosis was cultured from 27 (3.3%) of 816 pasteurized milk samples overall, 5 on Herrold's egg yolk medium and 22 by BACTEC cult...

  17. Factors affecting branch wound occlusion and associated decay following pruning – a case study with wild cherry (Prunus avium L.

    Directory of Open Access Journals (Sweden)

    Jonathan Sheppard

    2016-11-01

    Full Text Available Pruning wild cherry (Prunus avium L. is a common silvicultural practice carried out to produce valuable timber at a veneer wood quality. Sub-optimal pruning treatments can permit un-occluded pruning wounds to develop devaluing decay. The aim of this study is to determine relevant branch, tree and pruning characteristics affecting the occlusion process of pruning wounds. Important factors influencing occlusion time for an optimised pruning treatment for valuable timber production utilising wild cherry are derived. 85 artificially pruned branches originating from ten wild cherry trees were retrospectively analysed. Branch stub length, branch diameter and radial stem increment during occlusion were found to be significant predictors for occlusion time. From the results it could be concluded that for the long term success of artificial pruning of wild cherry it is crucial to (i keep branch stubs short (while avoiding damage to the branch collar, (ii to enable the tree to maintain significant radial growth after pruning, (iii to avoid large pruning wounds (>2.5 cm by removing steeply angled and fast growing branches at an early stage.

  18. Antibiotic resistances in Listeria monocytogenes and Salmonella enterica isolated from foods with animal origin Resistencias a antibióticos en Listeria monocytogenes y Salmonella enterica aislados de alimentos de origen animal

    OpenAIRE

    Baltasar Balsalobre Hernández; Joaquín Hernández-Godoy

    2004-01-01

    Extensive use of antibiotics in both human and animal health and in cattle production has generated resistant microorganisms to common antibiotics. Resistances spread caused by human and animal therapeutic is well known, but we know poorly frecuency of resistant bacteria in foods with animal origin and destinated to human consumers. In this paper, sensitivity to nineteen antibiotics was investigated in Listeria monocytogenes and Salmonella enterica strains isolated from foods with animal orig...

  19. Unexpected distribution of the fluoroquinolone-resistance gene qnrB in Escherichia coli isolates from different human and poultry origins in Ecuador.

    Science.gov (United States)

    Armas-Freire, Paulina I; Trueba, Gabriel; Proaño-Bolaños, Carolina; Levy, Karen; Zhang, Lixin; Marrs, Carl F; Cevallos, William; Eisenberg, Joseph N S

    2015-06-01

    Fluoroquinolone resistance can be conferred through chromosomal mutations or by the acquisition of plasmids carrying genes such as the quinolone resistance gene (qnr). In this study, 3,309 strains of commensal Escherichia coli were isolated in Ecuador from: (i) humans and chickens in a rural northern coastal area (n = 2368, 71.5%) and (ii) chickens from an industrial poultry operation (n = 827, 25%). In addition, 114 fluoroquinolone-resistant strains from patients with urinary tract infections who were treated at three urban hospitals in Quito, Ecuador were analyzed. All of the isolates were subjected to antibiotic susceptibility screening. Fluoroquinolone-resistant isolates (FRIs) were then screened for the presence of qnrB genes. A significantly higher phenotypic resistance to fluoroquinolones was determined in E. coli strains from chickens in both the rural area (22%) and the industrial operation (10%) than in strains isolated from humans in the rural communities (3%). However, the rates of qnrB genes in E. coli isolates from healthy humans in the rural communities (11 of 35 isolates, 31%) was higher than in chickens from either the industrial operations (3 of 81 isolates, 6%) or the rural communities (7 of 251 isolates, 2.8%). The occurrence of qnrB genes in human FRIs obtained from urban hospitals was low (1 of 114 isolates, 0.9%). These results suggested that the qnrB gene is more widely distributed in rural settings, where antibiotic usage is low, than in urban hospitals and industrial poultry operations. The role of qnrB in clinical resistance to fluoroquinolones is thus far unknown.

  20. New isolates of carnation Italian ringspot virus differ from the original one by having replication-associated proteins with a typical tombusvirus-like N-terminus and by inducing peroxisome- rather than mitochondrion-derived multivesicular bodies.

    Science.gov (United States)

    Koenig, Renate; Lesemann, Dietrich-Eckhardt; Pfeilstetter, Ernst

    2009-01-01

    Five new isolates of carnation Italian ringspot virus (CIRV) from cherry trees, Gypsophila and surface water differ from the original carnation isolate (CIRV-car) and also from Pelargonium necrotic spot virus (PelNSV) by having an ORF 1/ORF1-RT with a typical tombusvirus-like 5'end and by inducing the formation of peroxisome- rather than mitochondrion-derived multivesicular bodies (MVBs). This supports with natural isolates earlier conclusions reached by others with artificially produced hybrid viruses that the 5'end of ORF 1 determines from which organelle the MBVs will be derived. CIRV-car might have resulted from a natural recombination event with genome elements of a PelNSV-like virus.

  1. Comparative analysis of the susceptibility to biocides and heavy metals of extended-spectrum β-lactamase-producing Escherichia coli isolates of human and avian origin, Germany.

    Science.gov (United States)

    Deus, Daniela; Krischek, Carsten; Pfeifer, Yvonne; Sharifi, Ahmad Reza; Fiegen, Ulrike; Reich, Felix; Klein, Guenter; Kehrenberg, Corinna

    2017-02-08

    A total of 174 extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolates collected from humans (n=140) and healthy broiler chickens (n = 34) was included in the study. The MIC values of alkyl diaminoethyl glycin hydrochloride, benzethonium chloride, benzalkonium chloride, chlorhexidine, acriflavine, copper sulfate, silver nitrate and zinc chloride were determined by the broth microdilution method. Significant differences in MIC distributions were found between human and avian isolates and between CTX-M-, SHV- and TEM-type ESBL E. coli for chlorhexidine, silver nitrate, zinc chloride and copper sulfate by statistical analysis. Isolates with reduced susceptibility were investigated for the presence and localization of tolerance-mediating genes by PCR analysis and Southern blotting. The genes emrE, mdfA, sugE(c), cueO, copA, zntA and zitB were commonly present in isolates with elevated MICs, while the genes qacE∆1, qacF, qacH, sugE(p), cusC and pcoA, were less prevalent. In several isolates, a plasmid localization of the genes qacE∆1, qacF, qacH and sugE(p) on large plasmids >20 kb was detected.

  2. Disseminated mycobacteriosis manifesting as paraplegia in two Parma wallabies (Macropus parma) naturally exposed to Mycobacterium avium.

    Science.gov (United States)

    Robveille, Cynthia; Albaric, Olivier; Gaide, Nicolas; Abadie, Jérome

    2015-11-01

    Two captive female Parma wallabies (Macropus parma) died after a history of flaccid paraplegia. On postmortem examination, granulomatous and suppurative osteomyelitis involving the left ischium and the lumbosacral region, with meningeal extension at the cauda equina, and caseonecrotic mastitis were the most significant changes. Multiple small nodules in the liver and spleen, and an enlargement of some lymph nodes with central caseous necrosis were also observed. Microscopically, a disseminated granulomatous inflammation with numerous multinucleate giant cells was seen. Numerous acid-fast bacilli were detected in macrophages, in multinucleated giant cells, and free in the central necrosis and suppurative exudate. After culture, polymerase chain reaction assays were carried out to detect the 65-kDa heat shock protein (Hsp65) and insertion sequences (IS)1245 and IS900. The causative agent was identified as Mycobacterium avium subsp. avium.

  3. Mycobacterium avium subspecies paratuberculosis infects and multiplies in enteric glial cells

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To establish the role of enteric glial cells duringinfection with Mycobacterium avium subspeciesparatuberculosis (MAP) in Crohn's disease.METHODS: In order to establish the role of enteric glial cells during infection with M. avium subspecies paratuberculosis (MAP) in Crohn's disease, Map adhesion experiments on enteric glial cells were performed as well as expression analysis of Map sigma factors during infection.RESULTS: In this study, for the first time, we found a high affinity of MAP to enteric glial cells and we analyzed the expression of MAP sigma factors under different conditions of growth.CONCLUSION: The fact that Map showed a high affinity to the glial cells raises concerns about the complicated etiology of the Crohn's disease. Elucidation of the mechanisms whereby inflammation alters enteric neural control of gut functions may lead to novel treatments for Crohn's disease.

  4. The thioamides methimazole and thiourea inhibit growth of M. avium Subspecies paratuberculosis in culture.

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    Robert J Greenstein

    Full Text Available BACKGROUND: Thyrotoxicosis is conceptualized as an "autoimmune" disease with no accepted infectious etiology. There are increasingly compelling data that another "autoimmune" affliction, Crohn disease, may be caused by Mycobacterium avium subspecies paratuberculosis (MAP. Like M. tb, MAP is systemic. We hypothesized that some cases of thyrotoxicosis may be initiated by a MAP infection. Because other thioamides treat tuberculosis, leprosy and M. avium complex, we hypothesized that a mode of action of some thioamide anti-thyrotoxicosis medications may include MAP growth inhibition. METHODS: The effect of the thioamides, thiourea, methimazole and 6-propo-2-thiouracil (6-PTU were studied in radiometric Bactec culture, on ten strains of three mycobacterial species (six of MAP, two of M. avium and two of M. tb. complex. Data are presented as "cumulative growth index," (cGI or "percent decrease in cumulative GI" (%-DeltacGI. PRINCIPAL FINDINGS: Methimazole was the most effective thioamide at inhibiting MAP growth. At 128microg/ml: MAP UCF-4; 65%-DeltacGI & MAP ATCC 19698; 90%-DeltacGI. Thiourea inhibited MAP "Ben" maximally; 70%-DeltacGI. Neither methimazole nor thiourea inhibited M. avium or M. tb. at the doses tested. 6-PTU has no inhibition on any strain studied, although a structurally analogous control, 5-PTU, was the most inhibitory thioamide tested. SIGNIFICANCE: We show inhibition of MAP growth by the thioamides, thiourea and methimazole in culture. These data are compatible with the hypothesis that these thioamides may have anti-prokaryotic in addition to their well-established eukaryotic actions in thyrotoxic individuals.

  5. Microaggregate-associated protein involved in invasion of epithelial cells by Mycobacterium avium subsp. hominissuis

    OpenAIRE

    Babrak, Lmar; Danelishvili, Lia; Sasha J. Rose; Bermudez, Luiz E.

    2015-01-01

    The environmental opportunistic pathogen Mycobacterium avium subsp hominissuis (MAH), a member of the nontuberculous mycobacteria (NTM) cluster, causes respiratory as well as disseminated disease in patients such as those with chronic respiratory illnesses or AIDS. Currently, there is no effective method to prevent NTM respiratory infections. The formation of mycobacterial microaggregates comprises of phenotypic changes that lead to efficient adherence and invasion of the respiratory mucosa i...

  6. Mycobacterium avium complex augments macrophage HIV-1 production and increases CCR5 expression

    OpenAIRE

    1998-01-01

    Infection with HIV-1 results in pronounced immune suppression and susceptibility to opportunistic infections (OI). Reciprocally, OI augment HIV-1 replication. As we have shown for Mycobacterium avium complex (MAC) and Pneumocystis carinii, macrophages infected with opportunistic pathogens and within lymphoid tissues containing OI, exhibit striking levels of viral replication. To explore potential underlying mechanisms for increased HIV-1 replication associated with coinfection, blood monocyte...

  7. Inhibitory activity of lipid fractions myobacterium avium complex against macrophage respiratory burst

    OpenAIRE

    Shimizu, Toshiaki; 冨岡, 治明

    1998-01-01

    To explore possible mechanisms of the resistance of Mycobacterium avium complex (MAC) intracellular parasites to the antimicrobial activity of macrophages (MΦs), effects of the lipid components of these parasites on the MΦ respiratory burst were investigated. In this study, the MΦ respiratory burst was measured by luminoldependent chemiluminescence generated through the peroxidase-mediated halogenation reaction in murine peritoneal MΦs in response to phorbol myristate acetate (PMA) triggering...

  8. Molecular epidemiology and antimicrobial susceptibility of Campylobacter jejuni and Campylobacter coli isolates of poultry, swine, and cattle origin collected from slaughterhouses in Hungary.

    Science.gov (United States)

    Schweitzer, Nóra; Dán, Ádám; Kaszanyitzky, Éva; Samu, Péterné; Tóth, Ádám György; Varga, János; Damjanova, Ivelina

    2011-06-01

    Campylobacter spp. are the most common cause of bacterial enteritis in Hungary, and the aim of this study was to identify the distribution, genotypes, and antimicrobial susceptibility of Campylobacter species in the most important food-producing animals at the time of slaughter during 2008 and 2009. Of 1,110 samples, 266 were identified as Campylobacter coli (23.9%) and 143 as C. jejuni (12.9%) by real-time PCR. Resistance to enrofloxacin-ciprofloxacin and nalidixic acid was significant, especially in C. jejuni (73.3%) and C. coli (77.2%) from broilers. Higher erythromycin (P = 0.043) and tetracycline (P = 1.865e-14) resistance rates were found among C. coli isolates (9.7 and 74.1%, respectively) than among C. jejuni isolates (3.1 and 36.6%, respectively). A total of 47 fla short variable region sequences were identified among 73 selected C. coli and C. jejuni isolates, with 35 fla types detected only once. At the nucleotide level, fla types A66 and A21 were the most common. Using the pulsed-field gel electrophoresis method, 66% of strains exhibited unique profiles after Sma I digestion. Forty-two isolates assigned to 18 Sma I clusters were further typed by Kpn I, and of these, 24 were assigned to 10 Kpn I clusters. For isolates in five Kpn I clusters, epidemiological links were observed. Stable C. jejuni and C. coli clones were detected, indicating that further studies involving broiler and human isolates need to be conducted to elucidate the importance of these stable clones in human infections.

  9. Radioprotection of Swiss albino mice by Prunus avium with special reference to hematopoietic system.

    Science.gov (United States)

    Sisodia, Rashmi; Singh, Smita; Mundotiya, Chaturbhuj; Meghnani, Ekta; Srivastava, Preeti

    2011-01-01

    Prunus avium (family Rosaceae) has been used ethnomedicinally for the treatment of many diseases,but its radioprotective efficacy has hardly been explored. Presence of high anthocyanin content and phenolic compound with good antioxidative capacity has been reported by researchers. Its radioprotective effect against 5, 7, 10, and 12 Gygamma radiation was evaluated by 30 day survival assay. Regression analysis yielded LD(50/30) 5.81 and 9.43Gy for irradiated only and (P. avium fruit extract) PAE + radiation groups, respectively. The dose reduction factor was computed as 1.62. For biochemical and hematological studies, Swiss albino mice were divided into four groups: (i) control (vehicle treated), (ii) PAE treated (450 mg kg/day for 15 consequetive days), (iii) irradiated (5 Gy), and (4) PAE + irradiated. The irradiation of animals resulted in a significant elevation of lipid peroxidation and depletion in glutathione and protein levels in blood serum and spleen, which could be significantly checked by administration of PAE. Radiation-induced deficit in blood sugar, cholesterol, and hematological constituents could also be modulated by supplementation of PAE before and after irradiation. The possible prophylactic and therapeutic action noted by P. avium against radiation induced metabolic disorders may be due to synergistic action of various antioxidants, minerals, vitamins, etc., present in the fruit. Further mechanistic studies aimed at identifying the role of major ingredients in the extract are needed.

  10. Acute toxicity effects of Prunus avium fruit extract and selection of optimum dose against radiation exposure.

    Science.gov (United States)

    Sisodia, Rashmi; Sharma, K; Singh, Smita

    2009-01-01

    The objective of the study was to evaluate the acute toxicity of different doses of the methanolic extract of the fruit pulp of Prunus avium (family Rosaceae), which is used ethno-medicinally for the treatment of various diseases, and to find out the optimal dose of Prunus avium extract against 10 Gy gamma-radiation exposure. To test acute toxicity in mice, different doses of PAE (Prunus avium fruit extract) were given orally for 15 consecutive days, after which the animals were observed for another 15 days; the LD50/15 of the methanolic extract was calculated to be 4.947 gm/kg body weight (b.wt). In optimum dose selection against radiation exposure, oral administration of 450 mg/kg b.wt/d of PAE for 15 consecutive days before exposure to 10 Gy of gamma-radiation was found to afford maximum protection in terms of body weight and survivability of the mice in comparison to other doses.

  11. The Effect of Mycobacterium avium Complex Infections on Routine Mycobacterium bovis Diagnostic Tests

    Directory of Open Access Journals (Sweden)

    Claire Barry

    2011-01-01

    Full Text Available Bovine tuberculosis (bTB is diagnosed in naturally infected populations exposed to a wide variety of other pathogens. This study describes the cell-mediated immune responses of cattle exposed to Mycobacterium avium subspecies paratuberculosis (Map and Mycobacterium avium subspecies avium with particular reference to routine antefmortem Mycobacterium bovis diagnostic tests. The IFN-γ released in response to stimulated blood was found to peak later in the Map-exposed group and was more sustained when compared to the Maa-exposed group. There was a very close correlation between the responses to the purified protein derivatives (PPD used for stimulation (PPDa, PPDb, and PPDj with PPDa and PPDj most closely correlated. On occasion, in the Map-infected cattle, PPDb-biased responses were seen compared to PPDa suggesting that some Map-infected cattle could be misclassified as M. bovis infected using this test with these reagents. This bias was not seen when PPDj was used. SICCT results were consistent with the respective infections and all calves would have been classed skin test negative.

  12. Contemporary pollen flow, characterization of the maternal ecological neighbourhood and mating patterns in wild cherry (Prunus avium L.).

    Science.gov (United States)

    Cottrell, J E; Vaughan, S P; Connolly, T; Sing, L; Moodley, D J; Russell, K

    2009-08-01

    Conversion of lowland woodland to agricultural land and resulting fragmentation in Britain has been ongoing since Neolithic times. To counteract this decline, plantations of native species, often based on non-British planting stock, have been established. This may ultimately be detrimental to the integrity of the native gene pool. We explore the genetic and ecological factors influencing the success of components of the local pollen pool, including the effect of a non-native planting on an ancient woodland population of wild cherry. Wild cherry exhibits gametophytic self-incompatibility (GSI) and vegetative reproduction, both of which may be determinants of paternal success. The majority (61%) of the successful pollen originated from within the study site with a maximum pollen transfer distance of 694 m. There was a distinct departure from random mating, with over half the successful pollen originating from trees which occur within 100 m of the mother tree. Self-incompatibility, clonality, tree size and proximity to the mother tree were all found to influence paternal success. Kinship of pollen gametes within a maternal progeny was highest when a mother tree was surrounded by a large number of ramets of a single, compatible clone consisting of large, adult trees. Although the contribution from the non-native plantation is currently low, it is likely that this will increasingly contribute to the progeny of the adjacent ancient population as it matures. The results clearly show that in self-incompatible species, such as P. avium, close neighbours may be pollinated by very different components of the local pollen pool.

  13. Isolation and characterization of a distinct duck-origin goose parvovirus causing an outbreak of duckling short beak and dwarfism syndrome in China.

    Science.gov (United States)

    Chen, Shilong; Wang, Shao; Cheng, Xiaoxia; Xiao, Shifeng; Zhu, Xiaoli; Lin, Fengqiang; Wu, Nanyang; Wang, Jinxiang; Huang, Meiqing; Zheng, Min; Chen, Shaoying; Yu, Fusong

    2016-09-01

    Many mule duck and Cherry Valley duck flocks in different duck-producing regions of China have shown signs of an apparently new disease designated "short beak and dwarfism syndrome" (SBDS) since 2015. The disease is characterized by dyspraxia, weight loss, a protruding tongue, and high morbidity and low mortality rates. In order to characterize the etiological agent, a virus designated SBDSV M15 was isolated from allantoic fluid of dead embryos following serial passage in duck embryos. This virus causes a cytopathic effect in duck embryo fibroblast (DEF) cells. Using monoclonal antibody diagnostic assays, the SBDSV M15 isolate was positive for the antigen of goose parvovirus but not Muscovy duck parvovirus. A 348-bp (2604-2951) VP1gene fragment was amplified, and its sequence indicated that the virus was most closely related to a Hungarian GPV strain that was also isolated from mule ducks with SBDS disease. A similar disease was reproduced by inoculating birds with SBDSV M15. Together, these data indicate that SBDSV M15 is a GPV-related parvovirus causing SBDS disease and that it is divergent from classical GPV isolates.

  14. Comparison of cefoxitin disk diffusion test and mecA gene PCR results for methicillin resistance detection in Staphylococcus intermedius group isolates from canine origin in Brazil.

    Science.gov (United States)

    Penna, Bruno; Rabello, Renata F; Lilenbaum, Walter

    2014-01-01

    The study evaluated cefoxitin disk diffusion tests breakpoints and their correlation to mecA gene PCR results for detecting Methicillin-resistant Staphylococcus intermedius Group (MRSP) isolates from dogs in Brazil. Agreement using proposed breakpoint (resistant ≤ 30 mm) was encouraging. The current study reinforces that an epidemiological breakpoint can be established to predict presence of MRSP.

  15. Prevalence of genes for enterotoxins, toxic shock syndrome toxin 1 and exfoliative toxin among clinical isolates of Staphylococcus pseudintermedius from canine origin.

    Science.gov (United States)

    Yoon, Jang W; Lee, Gi-Jong; Lee, So-Young; Park, Chul; Yoo, Jong-Hyun; Park, Hee-Myung

    2010-10-01

    A total of 74 Staphylococcus pseudintermedius strains were isolated from the 99 clinical cases of canine pyoderma or chronic otitis in our veterinary teaching hospital during May 2006-February 2008. In this study, we examined the genetic distribution of staphylococcal pyogenic toxins such as staphylococcal enterotoxins A (sea), B (seb), C (sec), D (sed), E (see), and toxic shock syndrome toxin 1 (tst) as well as the previously characterized S. intermedius exfoliative toxin (siet) among those isolates. The polymerase chain reaction analyses with the toxin gene-specific primers revealed that 18 (24.3%) of 74 S. pseudintermedius isolates carried the sec genes, but none of the sea, seb, sed, see and tst genes. Further DNA sequencing analysis of the amplified sec genes revealed that they all belonged to the canine type C staphylococcal enterotoxin (SEC(canine) ) whose superantigenic activity has been demonstrated. In addition to the sec(canine) genes, our polymerase chain reaction results showed that all the 74 isolates carried the siet gene. Since both SEC(canine) and SIET toxins are known to be biologically active, it would be interesting to investigate how those toxins are involved in the pathogenesis of the canine diseases by S. pseudintermedius such as pyoderma or chronic otitis.

  16. Complete genome sequence of the biofilm-forming Curtobacterium sp. strain BH-2-1-1, isolated from lettuce (Lactuca sativa) originating from a conventional field in Norway.

    Science.gov (United States)

    Dees, Merete Wiken; Brurberg, May Bente; Lysøe, Erik

    2016-12-01

    Here, we present the 3,795,952 bp complete genome sequence of the biofilm-forming Curtobacterium sp. strain BH-2-1-1, isolated from conventionally grown lettuce (Lactuca sativa) from a field in Vestfold, Norway. The nucleotide sequence of this genome was deposited into NCBI GenBank under the accession CP017580.

  17. Variation in fumonisin and ochratoxin production associated with differences in biosynthetic gene content in Aspergillus niger and A. welwitschiae isolates from multiple crop and geographic origins

    Science.gov (United States)

    The fungi Aspergillus niger and A. welwitschiae are morphologically indistinguishable species used for industrial fermentation and for food and beverage production. The fungi also occur widely on food crops. Concerns about their safety have arisen with the discovery that some isolates of both specie...

  18. Molecular characterization of enterohemorrhagic Escherichia coli hemolysin gene (EHEC-hlyA)-harboring isolates from cattle reveals a diverse origin and hybrid diarrheagenic strains.

    Science.gov (United States)

    Askari Badouei, Mahdi; Morabito, Stefano; Najafifar, Arash; Mazandarani, Emad

    2016-04-01

    In the present study we investigated the occurrence of Escherichia coli strains harboring the gene encoding enterohemorrhagic E. coli hemolysin (EHEC-HlyA) in cattle and the association of this gene with various diarrheagenic E. coli (DEC) pathotypes. First, the bovine E. coli isolates were screened for EHEC-hlyA gene by PCR, and then they were characterized for the phylogenetic groups and the presence of the major virulence genes of different DEC pathotypes. In total, 25 virulence gene profiles were observed in 54 EHEC-hlyA+ isolates that reflect a considerable heterogeneity. The EHEC-hlyA+ strains were mostly associated with EHEC (72%), while only 7.4% were enteropathogenic E. coli (EPEC). We also showed the presence of estA gene of enterotoxigenic E. coli (ETEC) in 6 isolates (11.1%). Interestingly, two of the estA+ strains showed hybrid pathotypes with one carrying eae/estA (EPEC/ETEC), and the other one stx2/astA/estA (EHEC/ETEC). None of the isolates were related to enteroaggregative E. coli (EAggEC), enteroinvasive E. coli (EIEC), and necrotoxigenic E. coli (NTEC). The EHEC-plasmid encoded genes occurred in seven different combinations with EHEC-hlyA/saa/subA/espP being the most prevalent (46.3%). All stx-/eae+ strains carried O island 57 (OI-57) molecular marker(s) that may indicate these to be the progenitors of EHEC or strains losing stx. The most prevalent phylogroup was B1 (61.1%), but the most heterogeneous strains including the hybrid strains belonged to A phylogroup. Overall, our results indicate that cattle EHEC-hlyA encoding E. coli isolates consist of diverse diarrheagenic strains with the possible existence of hybrid pathotypes. Future studies are required to clarify the evolutionary aspects and clinical significance of these strains in humans and domestic animals.

  19. Characterization of Cronobacter spp. isolated from food of plant origin and environmental samples collected from farms and from supermarkets in the Czech Republic.

    Science.gov (United States)

    Vojkovska, Hana; Karpiskova, Renata; Orieskova, Maria; Drahovska, Hana

    2016-01-18

    The Cronobacter genus (previously known as Enterobacter sakazakii) comprises seven species (Cronobacter sakazakii, Cronobacter malonaticus, Cronobacter muytjensii, Cronobacter turicensis, Cronobacter dublinensis, Cronobacter universalis and Cronobacter condimenti)which cause serious infections in neonates and immunocompromised people.Most of the documented outbreaks of these bacteria have been associated with consumption of contaminated powdered infant formula. The plant environment is considered to be the natural habitat of these bacteria. Therefore, a total number of 563 samples of vegetables, fruit, water and environmental swabs were collected from local farms and supermarkets in the Czech Republic and investigated for the presence of Cronobacter spp. The obtained 45 isolates (8.0%) were further characterized by phenotyping (antimicrobial resistance, capsule and pigment production) and genotyping (fusA sequencing,MLST, PCR-serotyping) methods. Most of the Cronobacter isolates (42.2%) were identified as C. sakazakii, followed by C. turicensis (31.1%), C. dublinensis (22.2%), C. malonaticus (2.2%) and C. universalis (2.2%). The 25 identified sequence types, out of which 17 were unique for only one strain, indicated a high diversity of strains. C. sakazakii sequence type 4 (ST 4), which has been associated with many cases of meningitis, was isolated only in one case. A strong association of C. turicensis and C. dublinensis with the plant environment can be deduced from our results.

  20. Resposta imune específica de bovinos experimentalmente sensibilizados com inóculos inativados de Mycobacterium bovis e Mycobacterium avium Specific immune response of cattle to experimental sensibilization by inactivated Mycobacterium bovis and Mycobacterium avium

    Directory of Open Access Journals (Sweden)

    Robson F.C. Almeida

    2006-12-01

    Full Text Available O diagnóstico presuntivo da tuberculose bovina é baseado na análise da resposta imune celular a antígenos micobacterianos. Procedeu-se à simulação experimental de sensibilização por Mycobacterium bovis e Mycobacterium avium inativados em bovinos a fim de acompanhar a resposta imune a partir do teste cervical comparativo e da evolução da produção específica de interferon-gama, além de identificar a interferência de reações inespecíficas por M. avium nos resultados dos testes. Verificou-se que os animais desencadearam resposta de hipersensibilidade tardia contra os bacilos inativados, e que ambos os testes diagnósticos da tuberculose bovina foram eficientes na identificação dos animais sensibilizados com M. bovis e na discriminação das reações geradas pela inoculação dos bovinos com M. avium.The presumptive diagnosis of bovine tuberculosis is based on analysis of the immune response to micobacterial antigens. This experimental simulation of sensibilization by Mycobacterium bovis and Mycobacterium avium in cattle aimed to verify the immune response by both the cervical comparative test and the evolution of the specific production of gamma-interferon, and also to identify interference of unspecified reactions by M. avium on the test results. The results support that the experimental animals started a response of delayed hypersensitivity to the inactivated bacilli, and that both diagnostic tests for bovine tuberculosis were efficient for the identification of animals sensitized with M. bovis and for discrimination of reactions generated by inoculation of cattle with M. avium.

  1. Cloning and characterization of genomic DNA sequences of four self-incompatibility alleles in sweet cherry ( Prunus avium L.).

    Science.gov (United States)

    Wünsch, A; Hormaza, J I

    2004-01-01

    Gametophytic self-incompatibility (GSI) in sweet cherry is determined by a locus S with multiple alleles. In the style, the S-locus codifies for an allele-specific ribonuclease ( S-RNase) that is involved in the rejection of pollen that carries the same S allele. In this work we report the cloning and genomic DNA sequence analysis including the 5' flanking regions of four S-RNases of sweet cherry ( Prunus avium L., Rosaceae). DNA from the cultivars Ferrovia, Pico Colorado, Taleguera Brillante and Vittoria was amplified through PCR using primers designed in the conserved sequences of sweet cherry S-RNases. Two alleles were amplified for each cultivar and three of them correspond to three new S-alleles named S23, S24 and S25 present in 'Pico Colorado', 'Vittoria' and 'Taleguera Brillante' respectively. To confirm the identity of the amplified fragments, the genomic DNA of these three putative S-RNases and the allele S12 amplified in the cultivar Ferrovia were cloned and sequenced. The nucleotide and deduced amino-acid sequences obtained contained the structural features of rosaceous S-RNases. The isolation of the 5'-flanking sequences of these four S-RNases revealed a conserved putative TATA box and high similarity among them downstream from that sequence. However, similarity was low compared with the 5'-flanking regions of S-RNases from the Maloideae. S6- and S24-RNase sequences are highly similar, and most amino-acid substitutions among these two RNases occur outside the rosaceous hypervariable region (RHV), but within another highly variable region. The confirmation of the different specificity of these two S-RNases would help elucidate which regions of the S-RNase sequences play a role in S-pollen specific recognition.

  2. Linfadenite granulomatosa em suínos: linfonodos afetados e diagnóstico patológico da infecção causada por agentes do Complexo Mycobacterium avium Granulomatous lymphadenitis in swine: lymph nodes affected and pathologic diagnosis of the infection caused by Mycobacterium avium Complex agents

    Directory of Open Access Journals (Sweden)

    Nelson Morés

    2007-01-01

    Full Text Available Realizou-se um estudo com o objetivo de avaliar a distribuição de lesões granulomatosas nos linfonodos das carcaças de suínos abatidos e de comparar os métodos de isolamento do Complexo Mycobacterium avium (MAC, coloração de Ziehl-Neelsen (ZN, exames histopatológico e de imunohisto-química (IHQ para o diagnóstico da linfadenite granulomatosa causada por micobactérias do MAC. Foram utilizadas 431 amostras de linfonodos colhidos de 394 carcaças de suínos abatidos em 12 frigoríficos da Região Sul do Brasil, com o Serviço de Inspeção Federal (SIF. Os linfonodos que apresentavam lesões granulomatosas foram submetidos aos exames histológicos, ZN e IHQ com anticorpo monoclonal produzido com extrato celular de M. avium. A concordância entre os exames foi medida pelo teste Kappa, com nível de confiança de 95%. O exame macroscópico realizado pelo SIF identifica corretamente 90,3% das lesões granulomatosas, quando comparado com o exame histológico e a maioria das carcaças (92,5 apresentam lesões apenas nos linfonodos cadeia alimentar. O exame histológico confirmou a presença de lesões granulo-matosas em 90,3% dos linfonodos. As concordâncias entre os exames histopatológico e coloração de ZN (Kappa: 0,342 e de IHQ e o isolamento do MAC (Kappa: 0,102 foram baixas, porém alta entre os exames de IHQ e histológico com a presença de granulomas típicos nos linfonodos (Kappa: 0,973. O exame de IHQ associado ao exame histopatológico mostrou-se eficiente na identificação das lesões de linfadenite granulomatosa causadas pelo MAC.A study to evaluate the distribution of granulomatous lesions in the lymph nodes of swine carcasses was accomplished. The main objective was to compare the methods of isolation of mycobacteria of the Mycobacterium avium Complex (MAC, Ziehl-Neelsen (ZN staining, histopathological examination and immunohistochemistry (IHC for the diagnosis of granulomatous lymphadenitis caused by MAC. A total of 431

  3. Genetic relatedness among Campylobacter jejuni serotyped isolates of diverse origin as determined by numerical analysis of amplified fragment length polymorphism (AFLP) profiles

    DEFF Research Database (Denmark)

    Siemer, B.L.; Harrington, C.S.; Nielsen, E.M.;

    2004-01-01

    health. The remaining 30 groups contained isolates from humans, chickens and associated food products, cattle, sheep, turkeys, ostriches and/or dogs. Strains assigned to serotypes 2, 6/7, 11 and 12 formed major clusters at the 77.6% S-level. Most other serotypes did not form homogeneous clusters...... of the Study: The genetic relatedness among all 'Penner' serotypes of C. jejuni is assessed by AFLP analysis. In addition, further evidence of epidemic and host-specific clones of C. jejuni is provided....

  4. Phylogenetic analysis of Pomacea canaliculata isolates collected from rice fields in different origins of China by combined mitochondrial 12S and 16S genes.

    Science.gov (United States)

    Li, Xiao-Yan; Bian, Qing-Qing; Zhao, Guang-Hui

    2015-02-01

    To study the genetic relationships of Pomacea canaliculata collected from rice fields in China, the mitochondrial (mt) 12S and 16S of 9 P. canaliculata isolates from 5 southern provinces in China were sequenced and analyzed. The intra-specific sequence variations of P. canaliculata were 0-1.1% for 12S and 0--0.6% for 16S, while the inter-specific variations among common Pomacea species in mt 12S and 16S were 3.0-11.7% and 2.3-10.1%, respectively. Phylogenetic analysis based on combined sequences of mt 12S and 16S revealed complex genetic structure of P. canaliculata in China. Two phylogenetic groups of P. canaliculata were indicated in China with one group sistered to P. canaliculata isolates from USA, and two groups were even found in the same province. The phylogenetic relationships of Pomacea spp. also could be effectively inferred by combined sequences of mt 12S and 16S. These findings provided basic information for further study of population genetics and diffusion pattern of P. canaliculata in China as well as in the world.

  5. Antiplasmodial, cytotoxic activities and characterization of a new naturally occurring quinone methide pentacyclic triterpenoid derivative isolated from Salacia leptoclada Tul. (Celastraceae) originated from Madagascar

    Institute of Scientific and Technical Information of China (English)

    Fatiany Pierre Ruphin; Robijaona Baholy; Randrianarivo Emmanuel; Raharisololalao Amelie; Marie-Therese Martin; Ngbolua Koto-te-Nyiwa

    2013-01-01

    Objective:To validate scientifically the traditional use of Salacia leptoclada Tul. (Celastraceae) (S. leptoclada) and to isolate and elucidate the structure of the biologically active compound. Methods:Bioassay-guided fractionation of the acetonic extract of the stem barks of S. leptoclada was carried out by a combination of chromatography technique and biological experiments in viro using Plasmodium falciparum and P388 leukemia cell lines as models. The structure of the biologically active pure compound was elucidated by 1D and 2D NMR spectroscopy and mass spectrometry. Results:Biological screening of S. leptoclada extracts resulted in the isolation of a pentacyclic triterpenic quinone methide. The pure compound exhibited both in vitro a cytotoxic effect on murine P388 leukemia cells with IC50 value of (0.041±0.020) µg/mL and an antiplasmodial activity against the chloroquine-resistant strain FC29 of Plasmodium falciparum with an IC50 value of (0.052±0.030) µg/mL. Despite this interesting anti-malarial property of the lead compound, the therapeutic index was weak (0.788). In the best of our knowledge, the quinone methide pentacyclic triterpenoid derivative compound is reported for the first time in S. leptoclada. Conclusions:The results suggest that furthers studies involving antineoplastic activity is needed for the development of this lead compound as anticancer drug.

  6. Optimizing surveillance for South American origin influenza A viruses along the United States Gulf Coast through genomic characterization of isolates from blue-winged teal (Anas discors)

    Science.gov (United States)

    Ramey, Andy M.; Walther, Patrick; Link, Paul Karl; Poulson, Rebecca L.; Wilcox, Benjamin R.; Newsome, George M.; Spackman, Erica; Brown, J.; Stallknecht, David E.

    2016-01-01

    Relative to research focused on intercontinental viral exchange between Eurasia and North America, less attention has been directed towards understanding the redistribution of influenza A viruses (IAVs) by wild birds between North America and South America. In this study, we genomically characterized 45 viruses isolated from blue-winged teal (Anas discors) along the Texas and Louisiana Gulf Coast during March of 2012 and 2013, coincident with northward migration of this species from Neotropical wintering areas to breeding grounds in the United States and Canada. No evidence of South American lineage genes were detected in IAVs isolated from blue-winged teal supporting restricted viral gene flow between the United States and southern South America. However, it is plausible that blue-winged teal redistribute IAVs between North American breeding grounds and wintering areas throughout the Neotropics, including northern South America, and that viral gene flow is limited by geographical barriers further south (e.g. the Amazon Basin). Surveillance for the introduction of IAVs from Central America and northern South America into the United States may be further optimized through genomic characterization of viruses resulting from coordinated, concurrent sampling efforts targeting blue-winged teal and sympatric species throughout the Neotropics and along the United States Gulf Coast.

  7. The adipose tissue of origin influences the biological potential of human adipose stromal cells isolated from mediastinal and subcutaneous fat depots

    Directory of Open Access Journals (Sweden)

    Camilla Siciliano

    2016-09-01

    Full Text Available Indirect evidence suggests that adipose tissue-derived stromal cells (ASCs possess different physiological and biological variations related to the anatomical localization of the adipose depots. Accordingly, to investigate the influence of the tissue origin on the intrinsic properties of ASCs and to assess their response to specific stimuli, we compared the biological, functional and ultrastructural properties of two ASC pools derived from mediastinal and subcutaneous depots (thoracic compartment by means of supplements such as platelet lysate (PL and FBS. Subcutaneous ASCs exhibited higher proliferative and clonogenic abilities than mediastinal counterpart, as well as increased secreted levels of IL-6 combined with lower amount of VEGF-C. In contrast, mediastinal ASCs displayed enhanced pro-angiogenic and adipogenic differentiation properties, increased cell diameter and early autophagic processes, highlighted by electron microscopy. Our results further support the hypothesis that the origin of adipose tissue significantly defines the biological properties of ASCs, and that a homogeneric function for all ASCs cannot be assumed.

  8. Detection and organization of atrazine-degrading genetic potential of seventeen bacterial isolates belonging to divergent taxa indicate a recent common origin of their catabolic functions

    DEFF Research Database (Denmark)

    El Azhari, Najoi

    2007-01-01

    A collection of 17 atrazine-degrading bacteria isolated from soils was studied to determine the composition of the atrazine-degrading genetic potential (i.e. trzN, trzD and atz) and the presence of IS1071. The characterization of seven new atrazine-degrading bacteria revealed for the first time...... the trzN-atzBC gene composition in Gram-negative bacteria such as Sinorhizobium sp. or Polaromonas sp. Three main atrazine-degrading gene combinations (i) trzN–atzBC, (ii) atzABC–trzD and (iii) atzABCDEF were observed. The atz and trz genes were often located on plasmids, suggesting that plasmid...

  9. Immunogenicity of Mycobacterium avium subsp. paratuberculosis specific peptides for inclusion in a subunit vaccine against paratuberculosis

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Tollefsen, S.; Olsen, I.;

    efficacies. The main problem with available vaccines is their interference with surveillance and diagnosis of bovine tuberculosis and paratuberculosis. Our ultimate aim is to develop a subunit vaccine consisting of selected MAP peptides, which allow differentiation of infected from vaccinated animals. Here......Paratuberculosis in ruminants is caused by an infection with Mycobacterium avium subspecies paratuberculosis (MAP) and is a chronic disease characterized by granulomatous enteritis. Available vaccines against paratuberculosis consist of variations of whole bacteria with adjuvant showing various...... and are immunogenic. In the near future, a panel of selected peptides will be tested for efficacy as a vaccine against paratuberculosis with calves or goats experimentally infected with MAP....

  10. Cutaneous gallium uptake in patients with AIDS with mycobacterium avium-intracellulare septicemia

    Energy Technology Data Exchange (ETDEWEB)

    Allwright, S.J.; Chapman, P.R.; Antico, V.F.; Gruenewald, S.M.

    1988-07-01

    Gallium imaging is increasingly being used for the early detection of complications in patients with AIDS. A 26-year-old homosexual man who was HIV antibody positive underwent gallium imaging for investigation of possible Pneumocystis carinii pneumonia. Widespread cutaneous focal uptake was seen, which was subsequently shown to be due to mycobacterium avium-intracellulare (MAI) septicemia. This case demonstrates the importance of whole body imaging rather than imaging target areas only, the utility of gallium imaging in aiding the early detection of clinically unsuspected disease, and shows a new pattern of gallium uptake in disseminated MAI infection.

  11. Mean effective sensitivity for Mycobacterium avium subsp paratuberculosis infection in cattle herds

    DEFF Research Database (Denmark)

    Kirkeby, Carsten; Græsbøll, Kaare; Hisham Beshara Halasa, Tariq;

    2015-01-01

    Background: Mycobacterium avium subsp. paratuberculosis (MAP) infections in cattle are generally challenging to detect and cost-effective test strategies are consequently difficult to identify. MAP-specific antibody ELISAs for milk and serum are relatively inexpensive, but their utility...... within and between groups, and in some groups we found a bimodal distribution of MES. Dairy herds generally showed higher MES than non-dairy herds. Dairy herds in a control programme for paratuberculosis showed a MES similar to all other dairy herds from which animals >2.0 years were tested (both groups...

  12. Facts, myths and hypotheses on the zoonotic nature of Mycobacterium avium subspecies paratuberculosis.

    Science.gov (United States)

    Atreya, Raja; Bülte, Michael; Gerlach, Gerald-F; Goethe, Ralph; Hornef, Mathias W; Köhler, Heike; Meens, Jochen; Möbius, Petra; Roeb, Elke; Weiss, Siegfried

    2014-10-01

    Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of paratuberculosis (Johne's disease [JD]), a chronic granulomatous enteritis in ruminants. JD is one of the most widespread bacterial diseases of domestic animals with significant economic impact. The histopathological picture of JD resembles that of Crohn's disease (CD), a human chronic inflammatory bowel disease of still unresolved aetiology. An aetiological relevance of MAP for CD has been proposed. This and the ambiguity of other published epidemiological findings raise the question whether MAP represents a zoonotic agent. In this review, we will discuss evidence that MAP has zoonotic capacity.

  13. Characterisation of an ELISA detecting immunoglobulin G to Mycobacterium avium subsp. paratuberculosis in bovine colostrum

    DEFF Research Database (Denmark)

    Zervens, Lisa Marie-Louise; Nielsen, Søren Saxmose; Jungersen, Gregers

    2013-01-01

    Although colostrum has been used to detect specific immunoglobulin (Ig) G to Mycobacterium avium subsp. paratuberculosis (MAP) in cattle, confounding, non-specific reactions can be a problem. The objectives of this study were to determine the proportion of non-specific ELISA reactions in samples...... of colostrum taken between 0 and 4days-in-milk (DIM), and to assess the probability of an animal testing positive for MAP specific IgG over this time-period. Non-specific reactions were found in 3/365 (0.8%) of samples. The odds of an animal testing positive on day of calving were 130 times higher than at 4...

  14. Isolation, crystallization, and investigation of ribosomal protein S8 complexed with specific fragments of rRNA of bacterial or archaeal origin.

    Science.gov (United States)

    Tishchenko, S V; Vassilieva, J M; Platonova, O B; Serganov, A A; Fomenkova, N P; Mudrik, E S; Piendl, W; Ehresmann, C; Ehresmann, B; Garber, M B

    2001-09-01

    The core ribosomal protein S8 binds to the central domain of 16S rRNA independently of other ribosomal proteins and is required for assembling the 30S subunit. It has been shown with E. coli ribosomes that a short rRNA fragment restricted by nucleotides 588-602 and 636-651 is sufficient for strong and specific protein S8 binding. In this work, we studied the complexes formed by ribosomal protein S8 from Thermus thermophilus and Methanococcus jannaschii with short rRNA fragments isolated from the same organisms. The dissociation constants of the complexes of protein S8 with rRNA fragments were determined. Based on the results of binding experiments, rRNA fragments of different length were designed and synthesized in preparative amounts in vitro using T7 RNA-polymerase. Stable S8-RNA complexes were crystallized. Crystals were obtained both for homologous bacterial and archaeal complexes and for hybrid complexes of archaeal protein with bacterial rRNA. Crystals of the complex of protein S8 from M. jannaschii with the 37-nucleotide rRNA fragment from the same organism suitable for X-ray analysis were obtained.

  15. When Isolated at Full Receptivity, in Vitro Fertilized Wheat (Triticum aestivum, L. Egg Cells Reveal [Ca2+]cyt Oscillation of Intracellular Origin

    Directory of Open Access Journals (Sweden)

    Zsolt Pónya

    2014-12-01

    Full Text Available During in vitro fertilization of wheat (Triticum aestivum, L. in egg cells isolated at various developmental stages, changes in cytosolic free calcium ([Ca2+]cyt were observed. The dynamics of [Ca2+]cyt elevation varied, reflecting the difference in the developmental stage of the eggs used. [Ca2+]cyt oscillation was exclusively observed in fertile, mature egg cells fused with the sperm cell. To determine how [Ca2+]cyt oscillation in mature egg cells is generated, egg cells were incubated in thapsigargin, which proved to be a specific inhibitor of the endoplasmic reticulum (ER Ca2+-ATPase in wheat egg cells. In unfertilized egg cells, the addition of thapsigargin caused an abrupt transient increase in [Ca2+]cyt in the absence of extracellular Ca2+, suggesting that an influx pathway for Ca2+ is activated by thapsigargin. The [Ca2+]cyt oscillation seemed to require the filling of an intracellular calcium store for the onset of which, calcium influx through the plasma membrane appeared essential. This was demonstrated by omitting extracellular calcium from (or adding GdCl3 to the fusion medium, which prevented [Ca2+]cyt oscillation in mature egg cells fused with the sperm. Combined, these data permit the hypothesis that the first sperm-induced transient increase in [Ca2+]cyt depletes an intracellular Ca2+ store, triggering an increase in plasma membrane Ca2+ permeability, and this enhanced Ca2+ influx results in [Ca2+]cyt oscillation.

  16. Paragonimus heterotremus Chen and Hsia (1964), in Vietnam: a molecular identification and relationships of isolates from different hosts and geographical origins.

    Science.gov (United States)

    Le, Thanh H; Van De, Nguyen; Blair, David; McManus, Donald P; Kino, Hideto; Agatsuma, Takeshi

    2006-04-01

    Paragonimus heterotremus Chen and Hsia (1964), and paragonimiasis caused by this species is a newly detected disease in Vietnam. Twelve samples of Paragonimus (Platyhelminthes: Trematoda: Digenea: Paragonimidae) from different life-stages (eggs, miracidia, metacercariae, adults from natural and experimental hosts) and host species (crab, dog, cat and human) were collected in different geographical locations in Vietnam. DNA sequences were obtained from each for partial mitochondrial cytochrome c oxidase subunit 1 (cox1) (387 bp) and the entire second ribosomal internal transcribed spacer (ITS-2) (361 bp). The ITS-2 sequences were identical among all specimens, including those previously reported in GenBank. For cox1, there were sequence differences between specimens from Vietnam (four provinces, different locations) and those from Guangxi (China) and Saraburi (Thailand). Phylogenetic trees inferred from cox1 and ITS-2 sequences using sequence data for 15 P. heterotremus and for other Paragonimus spp. revealed that all P. heterotremus originating from Vietnam, Thailand and China form a distinct group. This information also confirms the identity of the Vietnamese specimens as P. heterotremus.

  17. Complete sequencing of the bla(NDM-1-positive IncA/C plasmid from Escherichia coli ST38 isolate suggests a possible origin from plant pathogens.

    Directory of Open Access Journals (Sweden)

    Tsuyoshi Sekizuka

    Full Text Available The complete sequence of the plasmid pNDM-1_Dok01 carrying New Delhi metallo-β-lactamase (NDM-1 was determined by whole genome shotgun sequencing using Escherichia coli strain NDM-1_Dok01 (multilocus sequence typing type: ST38 and the transconjugant E. coli DH10B. The plasmid is an IncA/C incompatibility type composed of 225 predicted coding sequences in 195.5 kb and partially shares a sequence with bla(CMY-2-positive IncA/C plasmids such as E. coli AR060302 pAR060302 (166.5 kb and Salmonella enterica serovar Newport pSN254 (176.4 kb. The bla(NDM-1 gene in pNDM-1_Dok01 is terminally flanked by two IS903 elements that are distinct from those of the other characterized NDM-1 plasmids, suggesting that the bla(NDM-1 gene has been broadly transposed, together with various mobile elements, as a cassette gene. The chaperonin groES and groEL genes were identified in the bla(NDM-1-related composite transposon, and phylogenetic analysis and guanine-cytosine content (GC percentage showed similarities to the homologs of plant pathogens such as Pseudoxanthomonas and Xanthomonas spp., implying that plant pathogens are the potential source of the bla(NDM-1 gene. The complete sequence of pNDM-1_Dok01 suggests that the bla(NDM-1 gene was acquired by a novel composite transposon on an extensively disseminated IncA/C plasmid and transferred to the E. coli ST38 isolate.

  18. Complete sequencing of the bla(NDM-1)-positive IncA/C plasmid from Escherichia coli ST38 isolate suggests a possible origin from plant pathogens.

    Science.gov (United States)

    Sekizuka, Tsuyoshi; Matsui, Mari; Yamane, Kunikazu; Takeuchi, Fumihiko; Ohnishi, Makoto; Hishinuma, Akira; Arakawa, Yoshichika; Kuroda, Makoto

    2011-01-01

    The complete sequence of the plasmid pNDM-1_Dok01 carrying New Delhi metallo-β-lactamase (NDM-1) was determined by whole genome shotgun sequencing using Escherichia coli strain NDM-1_Dok01 (multilocus sequence typing type: ST38) and the transconjugant E. coli DH10B. The plasmid is an IncA/C incompatibility type composed of 225 predicted coding sequences in 195.5 kb and partially shares a sequence with bla(CMY-2)-positive IncA/C plasmids such as E. coli AR060302 pAR060302 (166.5 kb) and Salmonella enterica serovar Newport pSN254 (176.4 kb). The bla(NDM-1) gene in pNDM-1_Dok01 is terminally flanked by two IS903 elements that are distinct from those of the other characterized NDM-1 plasmids, suggesting that the bla(NDM-1) gene has been broadly transposed, together with various mobile elements, as a cassette gene. The chaperonin groES and groEL genes were identified in the bla(NDM-1)-related composite transposon, and phylogenetic analysis and guanine-cytosine content (GC) percentage showed similarities to the homologs of plant pathogens such as Pseudoxanthomonas and Xanthomonas spp., implying that plant pathogens are the potential source of the bla(NDM-1) gene. The complete sequence of pNDM-1_Dok01 suggests that the bla(NDM-1) gene was acquired by a novel composite transposon on an extensively disseminated IncA/C plasmid and transferred to the E. coli ST38 isolate.

  19. Mutant-prevention concentration and mechanism of resistance in clinical isolates and enrofloxacin/marbofloxacin-selected mutants of Escherichia coli of canine origin.

    Science.gov (United States)

    Gebru, Elias; Choi, Myung-Jin; Lee, Seung-Jin; Damte, Dereje; Park, Seung Chun

    2011-10-01

    The antibacterial activity and selection of resistant bacteria, along with mechanisms of fluoroquinolone resistance, were investigated by integrating the static [MIC or mutant-prevention concentration (MPC)] and in vitro dynamic model approaches using Escherichia coli isolates from diseased dogs. Using the dynamic models, selected E. coli strains and enrofloxacin and marbofloxacin at a range of simulated area under concentration-time curve over a 24 h interval (AUC(24 h))/MIC ratios were investigated. Our results indicated increasing losses in susceptibility of E. coli upon continuous exposure to enrofloxacin and marbofloxacin in vitro. This effect was transferable to other fluoroquinolones, as well as to structurally unrelated drugs. Our results also confirmed an AUC(24 h)/MIC (AUC(24 h)/MPC)-dependent antibacterial activity and selection of resistant E. coli mutants, in which maximum losses in fluoroquinolone susceptibility occurred at simulated AUC(24 h)/MIC ratios of 40-60. AUC(24 h)/MPC ratios of 39 (enrofloxacin) and 32 (marbofloxacin) were considered protective against the selection of resistant mutants of E. coli. Integrating our MIC and MPC data with published pharmacokinetic information in dogs revealed a better effect of the conventional dosing regimen of marbofloxacin than that of enrofloxacin in restricting the selection of resistant mutants of E. coli. Target mutations, especially at codon 83 (serine to leucine) of gyrA, and overexpression of efflux pumps contributed to resistance development in both clinically resistant and in vitro-selected mutants of E. coli. We also report here a previously undescribed mutation at codon 116 of parC in two laboratory-derived resistant mutants of E. coli. Additional studies would determine the exact role of this mutation in fluoroquinolone susceptibility, as well as establish the importance of our findings in the clinical setting.

  20. Detection of Mycobacterium avium subsp. paratuberculosis from cattle and buffaloes in Egypt using traditional culture, serological and molecular based methods

    Directory of Open Access Journals (Sweden)

    G. S. Abdellrazeq

    2014-08-01

    Full Text Available Background: Johne's disease (JD caused by Mycobacterium avium subsp. paratuberculosis (MAP represents a real threat to the agriculture and dairy food industries and believed to be a potential public health problem. Signs of infection in ruminant include weight loss, diarrhea, decreased milk production, and eventually death. The definition of an infected animal based either on the presence of anti-MAP antibodies, or positive bacterial culture. No treatment for the disease exists and controlling the disease is difficult due to its long latent period. JD is a worldwide problem and multiple studies in many countries have been carried out to determine the prevalence of MAP infections. Although some primary non intensive studies confirm presence of JD in Egypt, the disease is currently neglected by the official Egyptian veterinary agencies. There is no official data, no national control program, and no used vaccine. Aim: This study aimed to evaluate three conventional diagnostic methods for MAP under the Egyptian circumstances with a general aim to determine the appropriate strategy to develop a JD control program. These methods were pooled fecal culture, humoral response and insertion sequence IS900 targets polymerase chain reaction (IS900 PCR. Materials and Methods: Fecal and serum samples (500 each were collected from Holstein-Friesian cattle and buffaloes housed in five Egyptian governorates. Fecal samples were examined for MAP on the basis of a strategic pooling procedure and performed on Herrold's Egg Yolk Agar Medium (HEYM. Smears were prepared from developed colonies and stained using a Ziehl-Neelsen (ZN technique. The identity of developed colonies was further confirmed by PCR analysis of IS900 sequence. Sera from both culture-positive and culture-negative animals were evaluated individually for humoral response. Results: Out of 50 pooled specimens, 34 (68% fecal cultures were positive for MAP. Serum positive samples of culture

  1. Prevention of Mycobacterium avium subsp. paratuberculosis (MAP) infection in Balb/c mice by feeding probiotic Lactobacillus acidophilus NP-51

    Science.gov (United States)

    The objective of this study was to examine effects of feeding Lactobacillus acidophilus strain NP51 to mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne’s disease. We hypothesized that feeding NP51 would increase Th-1 responses and decrease prog...

  2. Prevention of Mycobacterium avium subsp. paratuberculosis Infection in BALB/c Mice by Feeding Lactobacillus acidophilus Strain NP-51

    Science.gov (United States)

    The immune responses of 390 BALB/c mice fed the probiotic Lactobacillus acidophilus strain NP51® and infected with Mycobacterium avium subspecies paratuberculosis (MAP) were evaluated in a 6-month trial. Mice were randomized to nine treatment groups fed either viable- or heat-killed NP51 and inocula...

  3. The effects of progressing and nonprogressing Mycobacterium avium ssp. paratuberculosis infection on milk production in dairy cows

    NARCIS (Netherlands)

    Smith, Rebecca L.; Gröhn, Y. T.; Pradhan, A. K.; Whitlock, R. H.; Van Kessel, J. S.; Smith, J. M.; Wolfgang, D. R.; Schukken, Y. H.

    2016-01-01

    Longitudinal data from 3 commercial dairy herds in the northeast United States, collected from 2004 to 2011, were analyzed to determine the effect of Mycobacterium avium ssp. paratuberculosis (MAP) infection status and progression path on milk production. Disease status, as indicated by MAP test res

  4. Middle-aged to elderly women have a higher asymptomatic infection rate with Mycobacterium avium complex, regardless of body habitus.

    Science.gov (United States)

    Nishimura, Tomoyasu; Fujita-Suzuki, Yukiko; Mori, Masaaki; Carpenter, Stephen M; Fujiwara, Hiroshi; Uwamino, Yoshifumi; Tamizu, Eiko; Yano, Ikuya; Kawabe, Hiroshi; Hasegawa, Naoki

    2016-04-01

    Mycobacterium avium complex (MAC) pulmonary disease is prevalent in middle-aged to elderly women with a thin body habitus. By comparing the rate of serologically diagnosed asymptomatic MAC infection and body mass index among 1033 healthy subjects, we find that middle-aged to elderly women became infected with MAC, regardless of their body habitus.

  5. Novel antigens for detection of cell mediated immune responses to Mycobacterium avium subsp. paratuberculosis infection in cattle

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Aagaard, Claus; Nielsen, Søren Saxmose;

    2011-01-01

    Paratuberculosis is a chronic infection of the intestine of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Early stage MAP infection can be detected by measuring specific cell mediated immune responses, using the whole blood interferon-γ (IFN-γ) assay. Available IFN-γ assa...

  6. Potentiating day-old blood samples for detection of interferon-gamma responses following infection with Mycobacterium avium subsp. paratuberculosis

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Nielsen, Søren Saxmose; Jungersen, Gregers

    The interferon gamma (IFN-γ) test measuring specific cell-mediated immune responses in whole blood can be used for diagnosis at an early stage of Mycobacterium avium subsp. paratuberculosis (MAP) infection. A major obstacle for the practical use of IFN-γ testing is the recommended maximum 8 hour...

  7. Use of Novel Recombinant Antigens in the Interferon Gamma Assay for Detection of Mycobacterium Avium Subsp. Paratuberculosis Infection in Cattle

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Aagaard, Claus; Nielsen, Søren Saxmose;

    Early stage Mycobacterium avium subsp. paratuberculosis (MAP) infection can be detected by measuring antigen specific cell mediated immune responses by the interferon gamma (IFN-γ) assay. Available IFN-γ assay use purified protein derivate of Johnin (PPDj) leading to low specificity. The objectives...

  8. Use of novel recombinant antigens in the interferon gamma assay for detection of Mycobacterium avium subsp. paratuberculosis infection in cattle

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Aagaard, C.; Nielsen, Søren Saxmose;

    2012-01-01

    Early stage Mycobacterium avium subsp. paratuberculosis (MAP) infection may be detected by measuring antigen specific cell-mediated immune responses by the interferon-gamma (IFN-¿) assay. Available IFN-¿ assay use purified protein derivate of Johnin (PPDj) leading to low specificity. The objectives...

  9. Association between milk antibody and interferon-gamma responses in cattle from Mycobacterium avium subsp. paratuberculosis infected herds

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Jungersen, Gregers; Nielsen, Søren Saxmose

    2009-01-01

    Paratuberculosis is a chronic infection of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). It is possible to detect infection with paratuberculosis at different stages of disease by means of various diagnostic test strategies. The objective of the present study was to evalu...

  10. Culture and molecular method for detection of Mycobacterium tuberculosis complex and Mycobacterium avium subsp. paratuberculosis in milk and dairy products.

    Science.gov (United States)

    Messelhäusser, U; Kämpf, P; Hörmansdorfer, S; Wagner, B; Schalch, B; Busch, U; Höller, C; Wallner, P; Barth, G; Rampp, A

    2012-01-01

    A combined molecular and cultural method for the detection of the Mycobacterium tuberculosis complex (MTBC) and Mycobacterium avium subsp. paratuberculosis was developed and tested with artificially contaminated milk and dairy products. Results indicate that the method can be used for a reliable detection as a basis for first risk assessments.

  11. Characterization of the inflammatory phenotype of Mycobacterium avium subspecies paratuberculosis using a novel cell culture passage model

    Science.gov (United States)

    Understanding the pathogenic mechanisms and host responses to Johne’s disease, a chronic enteritis of ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP), is complicated by the multifaceted disease progression, late-onset host reaction, and the lack of ex vivo infection models ...

  12. Efficacy of novel lipid-formulated whole bacterial cell vaccines against Mycobacterium avium subsp paratuberculosis in sheep

    NARCIS (Netherlands)

    Griffin, J.F.T.; Hughes, A.D.; Liggett, S.; Farquhar, P.A.; Mackintosh, C.G.; Bakker, D.

    2009-01-01

    Mycobacterium avium subsp. paratuberculosis [MAP], the Causative agent of enteric Johne's disease, incurs significant economic losses to the livestock industry. Prophylactic vaccination can be employed as a control means, however mineral oil-based vaccines Currently in practice have limited efficacy

  13. Can Immune Response Mechanisms Explain the Fecal Shedding Patterns of Cattle Infected with Mycobacterium avium Subspecies paratuberculosis?

    NARCIS (Netherlands)

    Gesham, Magombedze; Shigetoshi, Eda; Koets, A.P.

    2016-01-01

    Johne’s disease (JD) is a chronic disease in ruminants and is caused by infection with Mycobacterium avium subspecies paratuberculosis (MAP). At late stages of the disease, MAP bacilli are shed via feces excretion and in turn create the potential for oral-fecal transmission. The role of the host imm

  14. Shedding of Mycobacterium avium subsp. paratuberculosis into milk and colostrum of naturally infected dairy cows over complete lactation cycles

    Science.gov (United States)

    The primary mode of transmission of Mycobacterium avium subsp. paratuberculosis (MAP) is fecal-oral. However, MAP is also shed into the milk and colostrum of infected cows. The objective of this study was to identify if an association exists between stage of MAP infection and days in lactation with ...

  15. Killing of mycobacterium avium by lactoferricin peptides: improved activity of arginine- and D-amino-acid-containing molecules

    NARCIS (Netherlands)

    Silva, T.; Magalhães, B.; Maia, S.; Gomes, P.; Nazmi, K.; Bolscher, J.G.M.; Rodriques, P.N.; Bastos, M.; Gomes, M.S.

    2014-01-01

    Mycobacterium avium causes respiratory disease in susceptible individuals, as well as disseminated infections in immunocompromised hosts, being an important cause of morbidity and mortality among these populations. Current therapies consist of a combination of antibiotics taken for at least 6 months

  16. CD4 T Cells From Intestinal Biopsies of Crohn's Disease Patients React to Mycobacterium avium subspecies paratuberculosis

    Science.gov (United States)

    The role of Mycobacterium avium subspecies paratuberculosis (MAP) in Crohn’s disease (CD) remains controversial. One issue that has been raised is the lack of data showing a cellular immune response to MAP. Earlier studies have mostly focused on responses in peripheral blood which have several limit...

  17. Occurrence of Mycobacterium avium subsp. paratuberculosis in milk at dairy cattle farms

    DEFF Research Database (Denmark)

    Okura, Hisako; Toft, Nils; Nielsen, Søren Saxmose

    2012-01-01

    Presence of Mycobacterium avium subsp. paratuberculosis (MAP) in milk for human consumption is a concern due to its possible relationship with Crohn’s disease in humans. Pasteurization effectively reduces the MAP load by four to five logs, but the efficacy depends on the MAP concentration, which...

  18. Differences in intermittent and continuous fecal shedding patterns between natural and experimental Mycobacterium avium subspecies paratuberculosis infections in cattle

    NARCIS (Netherlands)

    Mitchell, R.M.; Schukken, Ynte; Koets, A.P.; Weber, Maarten; Bakker, D.; Stabel, Judy; Whitlock, R.H.; Louzoun, Yoram

    2015-01-01

    The objective of this paper is to study shedding patterns of cows infected with Mycobacterium avium subsp. paratuberculosis (MAP). While multiple single farm studies of MAP dynamics were reported, there is not large scale meta-analysis of both natural and experimental infections. Large difference in

  19. Seed washing, exogenous application of gibberellic acid, and cold stratification enhance the germination of sweet cherry (Prunus avium L.) seed

    NARCIS (Netherlands)

    Javanmard, T.; Zamani, Z.; Keshavarz Afshar, R.; Hashemi, M.; Struik, P.C.

    2014-01-01

    Seed germination in sweet cherry (Prunus avium L.) is a slow and lengthy process which has delayed breeding efforts. In this study, seed from ripe fruit of the sweet cherry cultivar ‘Lambert’ were collected and, after removing the endocarp, various dormancy-breaking treatments such as seed washing,

  20. Rapid assessment of the viability of Mycobacterium avium subsp. paratuberculosis cells after heat treatment, using an optimized phage amplification assay.

    Science.gov (United States)

    Foddai, Antonio; Elliott, Christopher T; Grant, Irene R

    2010-03-01

    Thermal inactivation experiments were carried out to assess the utility of a recently optimized phage amplification assay to accurately enumerate viable Mycobacterium avium subsp. paratuberculosis cells in milk. Ultra-heat-treated (UHT) whole milk was spiked with large numbers of M. avium subsp. paratuberculosis organisms (10(6) to 10(7) CFU/ml) and dispensed in 100-microl aliquots in thin-walled 200-microl PCR tubes. A Primus 96 advanced thermal cycler (Peqlab, Erlangen, Germany) was used to achieve the following time and temperature treatments: (i) 63 degrees C for 3, 6, and 9 min; (ii) 68 degrees C for 20, 40, and 60 s; and (iii) 72 degrees C for 5, 10, 15, and 25 s. After thermal stress, the number of surviving M. avium subsp. paratuberculosis cells was assessed by both phage amplification assay and culture on Herrold's egg yolk medium (HEYM). A high correlation between PFU/ml and CFU/ml counts was observed for both unheated (r(2) = 0.943) and heated (r(2) = 0.971) M. avium subsp. paratuberculosis cells. D and z values obtained using the two types of counts were not significantly different (P > 0.05). The D(68 degrees C), mean D(63 degrees C), and D(72 degrees C) for four M. avium subsp. paratuberculosis strains were 81.8, 9.8, and 4.2 s, respectively, yielding a mean z value of 6.9 degrees C. Complete inactivation of 10(6) to 10(7) CFU of M. avium subsp. paratuberculosis/ml milk was not observed for any of the time-temperature combinations studied; 5.2- to 6.6-log(10) reductions in numbers were achieved depending on the temperature and time. Nonlinear thermal inactivation kinetics were consistently observed for this bacterium. This study confirms that the optimized phage assay can be employed in place of conventional culture on HEYM to speed up the acquisition of results (48 h instead of a minimum of 6 weeks) for inactivation experiments involving M. avium subsp. paratuberculosis-spiked samples.

  1. Molecular cloning, expression, and functional analysis of a predicted sulfotransferase STF9 from Mycobacterium avium.

    Science.gov (United States)

    Hossain, Md Murad; Moriizumi, Yuuji; Tanaka, Shotaro; Kimura, Makoto; Kakuta, Yoshimitsu

    2011-04-01

    Sulfotransferases catalyze the transfer of sulfate group from para-nitrophenyl sulfate (pNPS) or 3'-phosphoadenosine-5'-phosphosulfate (PAPS) onto acceptor molecules in the biosynthesis of sulfate esters. Human pathogenic mycobacteria are known to produce numerous sulfated molecules on their cell surface which have been implicated as important mediators in host-pathogen interactions. The open reading frame stf9, a predicted homologue of sulfotransferase in the Mycobacterium avium genomic data, was cloned and over expressed in Escherichia coli. The recombinant STF9 conserved the characteristic PAPS binding motif of sulfotransferase and was purified as a 44 kDa soluble protein which exhibited transfer of sulfate group from pNPS (K (m) 1.34 mM, V (max) 7.56 nmol/min/mg) onto 3'-phosphoadenosine-5'-phosphate (K (m) 0.24 mM, V (max) 10.36 nmol/min/mg). The recombinant STF9 protein was also capable of transferring sulfate group from PAPS onto certain acceptor substrates in E. coli, and showed binding affinity to the PAP-agarose resin, supporting the sulfotransferase activity of the recombinant STF9 protein. This is the first report of molecular evidence for sulfotransferase activity of a protein from M. avium. Mutation of Arg96 to Ala and Glu170 to Ala abolishes sulfotransferase activity, indicating the importance of Arg96 and Glu170 in STF9 activity catalysis.

  2. A photoelectrocatalytic process that disinfects water contaminated with Mycobacterium kansasii and Mycobacterium avium.

    Science.gov (United States)

    Brugnera, Michelle Fernanda; Miyata, Marcelo; Zocolo, Guilherme Julião; Leite, Clarice Queico Fujimura; Zanoni, Maria Valnice Boldrin

    2013-11-01

    Nontuberculous mycobacteria are resistant to conventional water treatment; indeed, they have been recovered from a wide variety of environmental sources. Here, we applied the photoelectrocatalytic technique using a Ti/TiO2-Ag photoanode to inactivate mycobacteria. For a mycobacteria population of 5 × 10(8) CFU mL(-1), we achieved 99.9 and 99.8% inactivation of Mycobacterium kansasii and Mycobacterium avium with rate constant of 6.2 × 10(-3) and 4.2 × 10(-3) min(-1), respectively, after 240 min. We compared the proposed method with the photolytic and photocatalytic methods. Using a mycobacteria population of 7.5 × 10(4) CFU mL(-1), the proposed Ti/TiO2-Ag photoanode elicited total mycobacteria inactivation within 3 min of treatment; the presence of Ag nanoparticles in the electrode provided 1.5 larger degradation rate constant as compared with the Ti/TiO2 anode (1.75 × 10(-2) for M. kansassi and 1.98 × 10(-2) for M. avium). We monitored the degradation of the metabolites released during cellular lysis by TOC removal, sugar release, chromatography, and mass spectrometry measurements; photoelectrocatalysis and Ti/TiO2-Ag photoanodes furnished the best results.

  3. Coexistent Pseudogout and Mycobacterium avium-intracellulare Septic Arthritis in a Patient with HIV and ESRD

    Directory of Open Access Journals (Sweden)

    Wais Afzal

    2016-01-01

    Full Text Available Pseudogout is a crystal-induced arthropathy characterized by the deposition of calcium pyrophosphate dihydrate (CPPD crystals in synovial fluid, menisci, or articular cartilage. Although not very common, this entity can be seen in patients with chronic kidney disease (CKD. Septic arthritis due to Mycobacterium avium-intracellulare (MAI is a rare entity that can affect immunocompromised patients such as those with acquired immunodeficiency syndrome (AIDS or those who are on immunosuppressive drugs. Here, we describe a 51-year-old female who presented with fever, right knee pain, swelling, warmth, and decreased range of motion for several days. The initial assessment was consistent with pseudogout, with negative bacterial and fungal cultures. However, due to high white blood cell (WBC count in the synovial fluid analysis, she was empirically started on intravenous (IV vancomycin and piperacillin-tazobactam and discharged on IV vancomycin and cefepime, while acid-fast bacilli (AFB culture was still in process. Seventeen days later, AFB culture grew Mycobacterium avium-intracellulare (MAI, and she was readmitted for relevant management. This case illustrates that septic arthritis due to MAI should be considered in the differential diagnosis of septic arthritis in immunocompromised patients.

  4. Hyperhydricity of Prunus avium shoots cultured on gelrite: a controlled stress response.

    Science.gov (United States)

    Franck, Thierry; Kevers, Claire; Gaspar, Thomas; Dommes, Jacques; Deby, Carol; Greimers, Roland; Serteyn, Didier; Deby-Dupont, Ginette

    2004-06-01

    Hyperhydricity is a physiological disorder frequently affecting shoots vegetatively propagated in vitro. Hyperhydric shoots are characterised by a translucent aspect due to a chlorophyll deficiency, a not very developed cell wall and a high water content. Hyperhydricity of Prunus avium shoots was expressed in vitro in one multiplication cycle by replacing the gelling agent agar (normal shoots: NS) by gelrite (hyperhydric shoots: HS). P. avium shoots evolving towards the hyperhydric state produced higher amounts of ethylene, polyamines (PAs) and proline, which are substances considered as stress markers. A higher activity of glutathione peroxidase (GPX; EC 1.11.1.9), involved in organic hydroperoxide elimination, suggested an increased production of these compounds in HS. The unchanged free fatty acid composition indicated no HS membrane damages compared to NS. The ploidy level of HS nuclei was not affected, but the bigger size and the lower percentage of nuclei during the S phase suggested a slowing down of the cell cycle. The results argued for a stress response of the HS, but no signs of oxidative damages of lipid membrane and nucleus were observed. The discussion points out paradoxical results in a classical analysis of stress and suggests an alternative way of defense mechanisms in HS, involving homeostatic regulation and controlled degradation processes to maintain integrity and vital functions of the cell.

  5. Antibody recognition to secreted proteins of Mycobacterium avium subsp. paratuberculosis in sera from infected ruminants.

    Science.gov (United States)

    Pradenas, M; Jara, M C; Hernández, N; Zambrano, A; Collins, M T; Kruze, J

    2009-09-18

    Two liquid culture media to obtain secreted proteins of Mycobacterium avium subsp. paratuberculosis at different incubation periods were evaluated. Middlebrook 7H9-OADC (7H9) and Watson-Reid (WR) broths were inoculated with a field strain of M. paratuberculosis and growth curves determined using nonlinear regression analysis. Most culture filtrate (CF) proteins were of low molecular weight and reacted strongly against sera from cultured-positive cases of paratuberculosis. CF proteins obtained in WR yielded a higher number of bands and were detected earlier than those obtained from 7H9. A high degree of variability in CF protein immunoreactivity was seen among infected animals. Sera from cattle with clinical paratuberculosis or heavy fecal shedders of M. paratuberculosis reacted more intensively and to more CF proteins than did sera from other infected cattle. Immunoblots showed differences in antibody binding to CF proteins when sera were absorbed with M. avium but not with others environmental mycobacteria. Immunoblots with sera from infected goats and a sheep showed reactivity with proteins of 32, 33 and 46kDa both before and after the sera were absorbed with M. phlei. Antibodies found in serum of infected deer reacted with CF proteins in a similar way as did for cattle. These results suggest that a pool of CF proteins of M. paratuberculosis could be good candidates as antigens for serodiagnosis of paratuberculosis.

  6. In vitro anti-Mycobacterium avium activities of quinolones: predicted active structures and mechanistic considerations.

    Science.gov (United States)

    Klopman, G; Li, J Y; Wang, S; Pearson, A J; Chang, K; Jacobs, M R; Bajaksouzian, S; Ellner, J J

    1994-08-01

    The relationship between the structures of quinolones and their anti-Mycobacterium avium activities has been previously derived by using the Multiple Computer-Automated Structure Evaluation program. A number of substructural constraints required to overcome the resistance of most of the strains have been identified. Nineteen new quinolones which qualify under these substructural requirements were identified by the program and subsequently tested. The results show that the substructural attributes identified by the program produced a successful a priori prediction of the anti-M. avium activities of the new quinolones. All 19 quinolones were found to be active, and 4 of them are as active or better than ciprofloxacin. With these new quinolones, the updated multiple computer-automated structure evaluation program structure-activity relationship analysis has helped to uncover additional information about the nature of the substituents at the C5 and C7 positions needed for optimal inhibitory activity. A possible explanation of drug resistance based on the observation of suicide inactivation of bacterial cytochrome P-450 by the cyclopropylamine moiety has also been proposed and is discussed in this report. Furthermore, we confirm the view that the amount of the uncharged form present in a neutral pH solution plays a crucial role in the drug's penetration ability.

  7. Mycobacterium avium Subspecies paratuberculosis Recombinant Proteins Modulate Antimycobacterial Functions of Bovine Macrophages.

    Science.gov (United States)

    Bannantine, John P; Stabel, Judith R; Laws, Elizabeth; D Cardieri, Maria Clara; Souza, Cleverson D

    2015-01-01

    It has been shown that Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) activates the Mitogen Activated Protein Kinase (MAPK) p38 pathway, yet it is unclear which components of M. paratuberculosis are involved in the process. Therefore, a set of 42 M. paratuberculosis recombinant proteins expressed from coding sequences annotated as lipoproteins were screened for their ability to induce IL-10 expression, an indicator of MAPKp38 activation, in bovine monocyte-derived macrophages. A recombinant lipoprotein, designated as MAP3837c, was among a group of 6 proteins that strongly induced IL-10 gene transcription in bovine macrophages, averaging a 3.1-fold increase compared to non-stimulated macrophages. However, a parallel increase in expression of IL-12 and TNF-α was only observed in macrophages exposed to a subset of these 6 proteins. Selected recombinant proteins were further analyzed for their ability to enhance survival of M. avium within bovine macrophages as measured by recovered viable bacteria and nitrite production. All 6 IL-10 inducing MAP recombinant proteins along with M. paratuberculosis cells significantly enhanced phosphorylation of MAPK-p38 in bovine macrophages. Although these proteins are likely not post translationally lipidated in E. coli and thus is a limitation in this study, these results form the foundation of how the protein component of the lipoprotein interacts with the immune system. Collectively, these data reveal M. paratuberculosis proteins that might play a role in MAPK-p38 pathway activation and hence in survival of this organism within bovine macrophages.

  8. Capsular arabinomannans from Mycobacterium avium with morphotype-specific structural differences but identical biological activity.

    Science.gov (United States)

    Wittkowski, Manon; Mittelstädt, Jessica; Brandau, Sven; Reiling, Norbert; Lindner, Buko; Torrelles, Jordi; Brennan, Patrick J; Holst, Otto

    2007-06-29

    The capsules of two colony morphotypes of Mycobacterium avium strain 2151 were investigated, i.e. the virulent smooth-transparent (SmT1) and the nonvirulent smooth-opaque (SmO) types. From both morphotypes we separated a nonacylated arabinomannan (AM) from an acylated polysaccharide fraction by affinity chromatography, of which the AMs were structurally characterized. The AMs from the virulent morphotype, in contrast to that from the nonvirulent form, possessed a larger mannan chain and a shorter arabinan chain. Incubation of murine bone marrow-derived macrophages and human dendritic cells showed that the acylated polysaccharide fractions were potent inducers of tumor necrosis factor-alpha, interleukin-12, and interleukin-10 compared with nonacylated AMs, which led to only a marginal cytokine release. Further in vitro experiments showed that both the acylated polysaccharide fractions and the nonacylated AMs were able to induce in vitro anti-tumor cytotoxicity of human peripheral blood mononuclear cells. Thus, morphotype-specific structural differences in the capsular AMs of M. avium do not correlate with biological activity; however, their acylation is a prerequisite for effective stimulation of murine macrophages and human dendritic cells.

  9. Detection of Mycobacterium tuberculosis and Mycobacterium avium Complexes by Real-Time PCR in Bovine Milk from Brazilian Dairy Farms.

    Science.gov (United States)

    Bezerra, André Vinícius Andrade; Dos Reis, Emily Marques; Rodrigues, Rogério Oliveira; Cenci, Alexander; Cerva, Cristine; Mayer, Fabiana Quoos

    2015-05-01

    Foodborne diseases are a public health problem worldwide. The consumption of contaminated raw milk has been recognized as a major cause of transmission of bovine tuberculosis to humans. Other mycobacteria that may be present in raw milk and may cause diseases are those belonging to the Mycobacterium avium complex. In this study, molecular biology tools were applied to investigate raw milk contamination with Mycobacterium spp. in family dairy farms from Rio Grande do Sul, southern Brazil. Furthermore, different variables related to the source of the milk, herd characteristics, and management were evaluated for their effect on milk contamination. Five hundred and two samples were analyzed, of which 354 were from the Northwest region (102 farms with samples from 93 bulk tanks and 261 animals) and 148 from the South region of the state (22 farms with samples from 23 bulk tanks and 125 animals). Among them, 10 (1.99%) and 7 (1.39%) were positive for Mycobacterium tuberculosis (9 confirmed as Mycobacterium bovis) and M. avium complexes, respectively. There was no difference in the frequencies of positive samples between the regions or the sample sources. Of the positive samples, 4 were collected from a bulk tank (1 positive for M. avium and 3 for M. tuberculosis). Moreover, 1 sample was positive concomitantly for M. tuberculosis and M. avium complexes. On risk analysis, no variable was associated with raw milk contamination by M. tuberculosis complex species. However, washing the udders of all animals and drying them with paper towels were weakly classified as risk factors for M. avium contamination. Positive samples were obtained from both animals and bulk tanks, which emphasizes the importance of tuberculosis control programs and provides evidence that milk monitoring can be used as a control practice. Moreover, the findings of this study reinforce the need for awareness of the problems of raw milk consumption among the general population.

  10. Immunoregulatory effects of Taishan Pinus massoniana pollen polysaccharide on chicks co-infected with avian leukosis virus and Bordetella avium early in ovo.

    Science.gov (United States)

    Guo, Fanxia; Xue, Cong; Wu, Cun; Zhao, Xue; Qu, Tinghe; He, Xiaohua; Guo, Zhongkun; Zhu, Ruiliang

    2014-04-01

    In recent years, co-infection of chicken embryos with immunosuppressive viruses and bacteria occurs with an annually increasing frequency. Consequently, studies on new and safe immunoregulators, especially plant polysaccharides, have become a popular topic in the poultry industry. In the present study, we selected 300 specific pathogen free embryonated eggs, which were injected with subgroup B avian leukosis virus (ALV-B) and Bordetella avium (B. avium) to establish an artificial co-infection model. The chicks that hatched from these co-infected embryonated eggs were treated with Taishan Pinus massoniana pollen polysaccharide (TPPPS). Results indicated that relevant indices in the co-infection group were significantly lower than that in B. avium-only group. Furthermore, pathogenicity of B. avium was exacerbated, with the chicks exhibiting decreased body weights. The TPPPS groups exhibited gradual improvements in immune function and developmental status. Therefore, in terms of improving immunologic function and production performance, TPPPS could be used as immunoregulator for immune responses.

  11. Bacteremia with an iliopsoas abscess and osteomyelitis of the femoral head caused by Enterococcus avium in a patient with end-stage kidney disease.

    Science.gov (United States)

    Okada, Akira; Hangai, Mika; Oda, Toshimi

    2015-01-01

    A 70-year-old man on hemodialysis for end-stage kidney disease due to polycystic kidney disease presented with hip pain on extension and a high C-reactive protein level. Further examinations revealed an iliopsoas abscess and femoral head osteomyelitis caused by Enterococcus avium (E. avium) detected in blood and pus cultures. Complete resolution of the infection with ampicillin-resistant E. avium required six months of vancomycin therapy and two surgical drainage procedures. There have been no previous case reports in which both blood and abscess cultures confirmed E. avium infection. Careful attention should be paid to the detection of non-specific symptoms in patients on hemodialysis, with blood cultures being essential in such cases.

  12. Treatment with antibiotics is detrimental to the recovery of viable Mycobacterium avium subsp. paratuberculosis cultured from milk and colostrum of dairy cows

    Science.gov (United States)

    Antibiotic cocktails are frequently used as secondary decontaminants prior to the culture of Mycobacterium avium subsp. paratuberculosis (MAP). This study investigated whether secondary incubation with an antibiotic cocktail containing vancomycin, nalidixic acid, and amphotericin B after primary exp...

  13. Isolation of Nontuberculous Mycobacteria (NTM) from Household Water and Shower Aerosols in Patients with Pulmonary Disease Caused by NTM

    OpenAIRE

    Thomson, Rachel; Tolson, Carla; Carter, Robyn; Coulter, Chris; Huygens, Flavia; Hargreaves, Megan

    2013-01-01

    It has been postulated that susceptible individuals may acquire infection with nontuberculous mycobacteria (NTM) from water and aerosol exposure. This study examined household water and shower aerosols of patients with NTM pulmonary disease. The mycobacteria isolated from clinical samples from 20 patients included M. avium (5 patients), M. intracellulare (12 patients), M. abscessus (7 patients), M. gordonae (1 patient), M. lentiflavum (1 patient), M. fortuitum (1 patient), M. peregrinum (1 pa...

  14. [Hypotension from endocrine origin].

    Science.gov (United States)

    Vantyghem, Marie-Christine; Douillard, Claire; Balavoine, Anne-Sophie

    2012-11-01

    Hypotension is defined by a low blood pressure either permanently or only in upright posture (orthostatic hypotension). In contrast to hypertension, there is no threshold defining hypotension. The occurrence of symptoms for systolic and diastolic measurements respectively below 90 and 60 mm Hg establishes the diagnosis. Every acute hypotensive event should suggest shock, adrenal failure or an iatrogenic cause. Chronic hypotension from endocrine origin may be linked to adrenal failure from adrenal or central origin, isolated hypoaldosteronism, pseudohypoaldosteronism, pheochromocytoma, neuro-endocrine tumors (carcinoïd syndrome) or diabetic dysautonomia. Hypotension related to hypoaldosteronism associates low blood sodium and above all high blood potassium levels. They are generally classified according to their primary (hyperreninism) or secondary (hyporeninism) adrenal origin. Isolated primary hypoaldosteronisms are rare in adults (intensive care unit, selective injury of the glomerulosa area) and in children (aldosterone synthase deficiency). Isolated secondary hypoaldosteronism is related to mellitus diabetes complicated with dysautonomia, kidney failure, age, iatrogenic factors, and HIV infections. In both cases, they can be associated to glucocorticoid insufficiency from primary adrenal origin (adrenal failure of various origins with hyperreninism, among which congenital 21 hydroxylase deficiency with salt loss) or from central origin (hypopituitarism with hypo-reninism). Pseudohypoaldosteronisms are linked to congenital (type 1 pseudohypoaldosteronism) or acquired states of resistance to aldosterone. Acquired salt losses from enteric (total colectomy with ileostomy) or renal (interstitial nephropathy, Bartter and Gitelman syndromes…) origin might be responsible for hypotension and are associated with hyperreninism-hyperaldosteronism. Hypotension is a rare manifestation of pheochromocytomas, especially during surgical removal when the patient has not been

  15. Development of a rapid phage-based method for the detection of viable Mycobacterium avium subsp. paratuberculosis in blood within 48 h☆

    Science.gov (United States)

    Swift, Benjamin M.C.; Denton, Emily J.; Mahendran, Sophie A.; Huxley, Jonathan N.; Rees, Catherine E.D.

    2013-01-01

    The aim of this study was to develop a methodology to rapidly detect viable Mycobacterium avium subsp. paratuberculosis (MAP) in clinical blood samples. MAP cells spiked into commercially available blood were recovered using optimised peptide-mediated magnetic separation (PMMS) and detected using a phage-based method, and the identity of the cells detected confirmed using nested-PCR amplification of MAP signature sequences (IS900). The limit of detection was determined to be 10 MAP cells per ml of blood and was used to detect MAP present in clinical bovine blood samples. Using the PMMS-phage method there was no difference when detecting MAP from whole blood or from isolated buffy coat. MAP was detected in animals that were milk-ELISA positive (15 animals) by PMMS-phage and no MAP was detected in blood samples from an accredited Johne's disease free herd (5 animals). In a set of samples from one herd (10 animals) that came from animals with variable milk ELISA status, the PMMS-phage results agreed with the positive milk-ELISA results in all but one case. These results show that the PMMS-phage method can detect MAP present in naturally infected blood. Total assay time is 48 h and, unlike PCR-based detection tests, only viable cells are detected. A rapid method for detecting MAP in blood could further the understanding of disseminated infection in animals with Johne's disease. PMID:23811207

  16. Intracellular fate of Mycobacterium avium subspecies paratuberculosis in monocytes from normal and infected, interferon-responsive cows as determined by a radiometric method.

    Science.gov (United States)

    Zhao, B Y; Czuprynski, C J; Collins, M T

    1999-01-01

    The ability of Mycobacterium avium subsp. paratuberculosis to survive in bovine monocytes was studied using radiometric (BACTEC) culture, standard plate counting and microscopic counting of acid-fast stained monocyte monolayers. Results of microscopic counts sharply contrasted with results of viable counts determined both by plate counting and radiometric counting. We observed an early phase (the first 6 d after in vitro infection) of intracellular bacillary growth, followed by a later phase of mycobacteriostasis or killing (up to 12 d after in vitro infection) in monocytes from non-infected cows. The data suggest that multiplication and death of M. avium subsp. paratuberculosis occur simultaneously in bovine monocytes infected in vitro. Using the BACTEC method, we compared the ability of bovine monocytes from normal cows and cows infected with M. avium subsp. paratuberculosis and showing evidence of a strong Thl-like cellular immune response to ingest and inhibit the intracellular growth of M. avium subsp. paratuberculosis. There was a trend toward greater phagocytosis and faster killing of Mycobacterium avium subsp. paratuberculosis by monocytes from the infected, immune responder cows. However, the observed numbers of viable M. avium subsp. paratuberculosis at each time after monocyte infection were not significantly different between normal and infected cows.

  17. Effect of feeding heat-treated colostrum on risk for infection with Mycobacterium avium ssp. paratuberculosis, milk production, and longevity in Holstein dairy cows.

    Science.gov (United States)

    Godden, S M; Wells, S; Donahue, M; Stabel, J; Oakes, J M; Sreevatsan, S; Fetrow, J

    2015-08-01

    study conclusion was not different for animals originally fed HT (68.0%) versus FR (71.7%) colostrum. Although a previous study showed that feeding HT colostrum (60°C for 60min) produces short-term benefits, including improved passive transfer of IgG and reduced morbidity in the preweaning period, the current study found no benefit of feeding HT colostrum on long-term outcomes including risk for transmission of Mycobacterium avium ssp. paratuberculosis, milk production in the first and second lactation, and longevity within the herd.

  18. Efficacy of 'indigenous vaccine' using native 'Indian bison type' genotype of Mycobacterium avium subspecies paratuberculosis for the control of clinical Johne's disease in an organized goat herd.

    Science.gov (United States)

    Singh, K; Chandel, B S; Chauhan, H C; Dadawala, A; Singh, S V; Singh, P K

    2013-06-01

    Therapeutic efficacy of a new 'Indigenous vaccine' prepared from native highly pathogenic 'Indian Bison Type' genotype of Mycobacterium avium subspecies paratuberculosis (MAP) of goat origin has been evaluated with respect to control of clinical Johne's disease in naturally infected Mehsana breed of goat in North Gujarat. Fifty goats from Sheep and Goats Research Station, Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar, were randomly divided into 2 groups viz.,'Vaccinated'(n = 35) and 'Control'(n = 15). After vaccination, goats were monitored for physical condition, morbidity, mortality, body weights, shedding of MAP in feces, internal condition, gross lesions and humoral immune responses up to 120 days (at each interval of 30 days). At the end of 120 days trial, there was marked overall improvement in physical condition and body weights of vaccinated goats as compared to 'Control' goats. Vaccinated goats gained significantly (P vaccinated goats were positive for MAP DNA in faecal PCR and blood PCR before vaccination. However, all were found as negative at 120 days post vaccination (DPV). Overall vaccine exhibited effective in restriction of MAP infection and significant improvement in production parameters and reduction in mortality and morbidity due to JD. The trial in the herd will be continued.

  19. Production and proteomic characterisation of purified protein derivative from Mycobacterium avium subsp. paratuberculosis

    Directory of Open Access Journals (Sweden)

    Wynne James W

    2012-03-01

    Full Text Available Abstract Background Effective diagnosis of Johne's disease (JD, particularly at the stage of early subclinical infection, remains one of the greatest challenges for the control of JD worldwide. The IFN-γ test of cell mediated immunity is currently one of the most suitable diagnostics for subclinical infections, however a major limitation of this test is the lack of a standardised purified protein derivative (PPD antigen (also referred to as Johnin PPD or PPDj. While attempting to replace PPDj with more specific individual antigens is an attractive proposition, bacterial culture derived PPDj remains the most effective antigen preparation for the diagnosis of subclinical JD. It may be possible to increase the reproducibility and specificity of PPDj preparations by further characterising and standardising the PPDj production. Results Using a standardised protocol, five in-house preparations of PPDj were prepared from cultures of Mycobacterium avium subsp. paratuberculosis (MAP. Compared to PPDs obtained from other institutes/laboratories, these preparations appeared to perform similarly well in the IFN-γ test. Although the broad proteomic composition of all PPDj preparations was remarkably similar, the absolute abundance of individual proteins varied markedly between preparations. All PPDj preparations contained common immunogenic proteins which were also observed in PPD preparations from Mycobacterium avium subsp. avium (PPDa and Mycobacterium bovis (PPDb. Temporal difference in protein secretion of in vitro cultured MAP was observed between 20 and 34 weeks suggesting that the age of MAP culture used for PPDj preparations may markedly influence PPDj composition. Conclusions This study describes a protocol for the production of PPDj and its subsequent proteomic characterisation. The broad proteomic composition of different preparations of PPDj was, for the most part, highly similar. Compositional differences between PPDj preparations were found

  20. Clofazimine Prevents the Regrowth of Mycobacterium abscessus and Mycobacterium avium Type Strains Exposed to Amikacin and Clarithromycin

    Science.gov (United States)

    Ferro, Beatriz E.; Meletiadis, Joseph; Wattenberg, Melanie; de Jong, Arjan; van Soolingen, Dick; Mouton, Johan W.

    2015-01-01

    Multidrug therapy is a standard practice when treating infections by nontuberculous mycobacteria (NTM), but few treatment options exist. We conducted this study to define the drug-drug interaction between clofazimine and both amikacin and clarithromycin and its contribution to NTM treatment. Mycobacterium abscessus and Mycobacterium avium type strains were used. Time-kill assays for clofazimine alone and combined with amikacin or clarithromycin were performed at concentrations of 0.25× to 2× MIC. Pharmacodynamic interactions were assessed by response surface model of Bliss independence (RSBI) and isobolographic analysis of Loewe additivity (ISLA), calculating the percentage of statistically significant Bliss interactions and interaction indices (I), respectively. Monte Carlo simulations with predicted human lung concentrations were used to calculate target attainment rates for combination and monotherapy regimens. Clofazimine alone was bacteriostatic for both NTM. Clofazimine-amikacin was synergistic against M. abscessus (I = 0.41; 95% confidence interval [CI], 0.29 to 0.55) and M. avium (I = 0.027; 95% CI, 0.007 to 0.048). Based on RSBI analysis, synergistic interactions of 28.4 to 29.0% and 23.2 to 56.7% were observed at 1× to 2× MIC and 0.25× to 2× MIC for M. abscessus and M. avium, respectively. Clofazimine-clarithromycin was also synergistic against M. abscessus (I = 0.53; 95% CI, 0.35 to 0.72) and M. avium (I = 0.16; 95% CI, 0.04 to 0.35), RSBI analysis showed 23.5% and 23.3 to 53.3% at 2× MIC and 0.25× to 0.5× MIC for M. abscessus and M. avium, respectively. Clofazimine prevented the regrowth observed with amikacin or clarithromycin alone. Target attainment rates of combination regimens were >60% higher than those of monotherapy regimens for M. abscessus and M. avium. The combination of clofazimine with amikacin or clarithromycin was synergistic in vitro. This suggests a potential role for clofazimine in treatment regimens that warrants further

  1. DETEKSI MYCOBACTERIUM AVIUM SUBSPECIES PARATUBERCULOSIS PADA SUSU FORMULA LANJUTAN DI BOGOR [Detection of Mycobacterium avium subspecies paratuberculosis Formula Milk in Bogor

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    Widagdo Sri Nugroho1,2

    2008-06-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (MAP becomes a public health concern in developed countries which is usually associated to Crohn’s disease (CD in human. The disease shows similarities in clinical signs and pathology characteristic with John’s disease (JD in ruminants which is infected by MAP. Researchers in Europe, USA, and Australia detected MAP in their dairy products and showed the relationship among MAP, CD, and JD. Meanwhile Indonesia imported milk and milk products from those countries to cover the national demand. This situation keeps MAP as potential-problem in national dairy herd and human health in the future. The aim of this study was to detect MAP in the formla milk for todler. Fifty samples from five established milk producers were taken on August 2006 at the supermarket in Bogor. Two seperate diagnostic methods were used parallel in this study i.e.: polymerase chain reaction method (PCR with insertion sequence F 57 as the primer and the Mycobacterial Growth Indicator Tube (MGIT. Neither MAP grew in MGIT after 20 weeks of incubation period but 5 samples were found positive by nested PCR. Although there was no evidence weather MAP grew from the samples in this study, the comprehensive and sustainable studies on MAP should be carried out with more extensive and varied samples, as well as in human to provide data on MAP in Indonesia.

  2. Analysis of wild sweet cherry (Prunus avium L. germplasm diversity in south-east Serbia

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    Mratinić Evica

    2012-01-01

    Full Text Available Ten wild growing sweet cherry (Prunus avium L. genotypes from South-East Serbia with different fruit skin color were analyzed for its phenological, morphological and chemical traits. Agronomic evaluation of germplasm accessions revealed considerable diversity among different accessions for all the characters studied. The analysis of variance revealed significant differences among all genotypes for almost all examined properties. Cluster analysis showed adequate grouping of wild sweet cherry genotypes according to pomological characterization and distinguished them into two distinct groups. The first group had two subgroups and consisted of seven genotypes, while the second one included only three accessions. Despite of the significant differences among genotypes, the total concentration of phenols made a clear separation between the clusters. The level of genetic diversity in these wild sweet cherry genotypes is very high and therefore these trees are useful sources of variability for attributes studied and can be employed in further breeding programs or conservation.

  3. Genetic diversity in wild sweet cherries (Prunus avium) in Turkey revealed by SSR markers.

    Science.gov (United States)

    Ercisli, S; Agar, G; Yildirim, N; Duralija, B; Vokurka, A; Karlidag, H

    2011-06-21

    Wild sweet cherry (Prunus avium) trees are abundant in the northern part of Turkey, including the Coruh Valley. We analyzed 18 wild sweet cherry genotypes collected from diverse environments in the upper Coruh Valley in Turkey to determine genetic variation, using 10 SSR primers. These SSR primers generated 46 alleles; the number of alleles per primer ranged from 3 to 7, with a mean of 4.6. The primer PS12A02 gave the highest number of polymorphic bands (N = 7), while CPSCT010, UDAp-401 and UDAp-404 gave the lowest number (N = 3). Seven groups were separated in the dendrogram, although most of the genotypes did not cluster according to phenological and morphological traits. This level of genetic diversity in these wild sweet cherry genotypes is very high and therefore these trees would be useful as breeders for crosses between cultivated sweet cherry and wild genotypes.

  4. Detection of Mycobacterium avium subsp. paratuberculosis by a Direct In Situ PCR Method

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    Fernando Delgado

    2011-01-01

    Full Text Available In situ detection of Mycobacterium avium subsp. paratuberculosis is useful for diagnosis and research of paratuberculosis. The aim of this paper was to detect this agent in formalin-fixed, paraffin-embedded tissue samples by a direct in situ PCR. The technique was performed on ileum or ileocaecal lymph node samples from 8 naturally infected cattle and 1 healthy calf, by using p89 and p92 primers for amplification of IS900 sequence. Moderate positive signal was detected in all positive samples and not in negative control, but tissues resulted were affected in many cases due to the enzymatic treatment and the high temperature exposition. Although the technique was useful for Map detection, the signal was lower than immunohistochemistry probably because of the fixation process. In one case, signal was higher, which might be due to the detection of spheroplasts. Thus, the described method should be recommended when others resulted negative or for spheroplasts detection.

  5. Effect of pH on the in vitro potency of clarithromycin against Mycobacterium avium complex.

    OpenAIRE

    Truffot-Pernot, C; Ji, B; Grosset, J

    1991-01-01

    Employing 7H11 agar medium at pH 6.6, the MICs of clarithromycin for 50% (MIC50) and 90% (MIC90) of 19 strains of Mycobacterium avium complex were 8 and 16 micrograms/ml, respectively. However, the MICs were 2 to 3 log2 dilutions lower in the 7H11 medium adjusted to pH 7.4, and the MICs on 10% OADC (oleic acid-albumin-dextrose-catalase)-enriched Mueller-Hinton agar at pH 7.3 were also 2 log2 dilutions lower than those measured on 7H11 agar at pH 6.6. Therefore, clarithromycin is more active a...

  6. Seroprevalence of Mycobacterium avium subspecies paratuberculosis in Korean black goats (Capra hircus aegagrus).

    Science.gov (United States)

    Lee, Kyung Woo; Jung, Byeong Yeal; Moon, Oun Kyoung; Yang, Dong Kun; Lee, Su Hwa; Kim, Ji Yeon; Kweon, Chang Hee

    2006-12-01

    In total, 582 sera from 116 black goat herds were analyzed by a commercially available ELISA kit to monitor the seroprevalence of Mycobacterium avium subspecies paratuberculosis (Mpt) in Korean black goats (Capra hircus aegagrus). The mean number of goats sampled per herd was 5.11, 4.66, and 5.38 for the northern, central, and southern regions of Korea, respectively. The apparent regional prevalence of Mpt was estimated at 18.2-38.2% and 4.6-15.3% for herds and goats, respectively. The Mpt-positive goats were predominantly detected in the south (n=28), compared to either the northern (n=9) or central (n=11) regions (chi=14.459, P<0.05). Our findings indicate that Mpt is prevalent among the goat population, but regional variation exists.

  7. Isolamento de cepas de Mycobacterium avium em búfalos abatidos para consumo

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    Freitas José de Arimatéa

    2001-01-01

    Full Text Available Duas cepas micobacterianas, isoladas no parênquima pulmonar e linfonodo apical de búfalos abatidos para consumo, procedentes de criatórios localizados na Ilha de Marajó (PA e submetidas à identificação segundo ensaios recomendados para o gênero Mycobacterium, foram identificadas como pertencentes ao complexo Mycobacterium avium. Apresentaram-se considerações relativas à associação desses organismos com a Aids -- e o papel dos alimentos nessa associação --, discutindo-se o impacto que a condição de germes oportunistas das espécies desse complexo têm na pandemia do HIV, assim como o risco potencial representado pelas infecções produzidas nos animais.

  8. GENETIC DIVERSITY OF SOME IRANIAN SWEET CHERRY (PRUNUS AVIUM) CULTIVARS USING MICROSATELLITE MARKERS AND MORPHOLOGICAL TRAITS.

    Science.gov (United States)

    Farsad, A; Esna-Ashari, M

    2016-01-01

    The aim of this study was to characterize 23 important Iranian sweet cherry (Prunus avium) cultivars collected from different provinces of Iran and 1 foreign cultivar, which was used as control, considered for breeding programs by using 21 microsatellite markers and 27 morphological traits. In sweet cherry (Prunus avium) accessions, leaf, fruit, and stone morphological characters were evaluated during two consecutive years. The study revealed a high variability in the set of evaluated sweet cherry accessions. The majority of important correlations were determined among variables representing fruit and leaf size and variables related to color. Cluster analysis distinguished sweet cherry accessions into two distinct groups. Principal component analysis (PCA) of qualitative and quantitative morphological parameters explained over 86.59% of total variability in the first seven axes. In PCA, leaf traits such as leaf length and width, and fruit traits such as length, width, and weight, and fruit flesh and juice color were predominant in the first two components, indicating that they were useful for the assessment of sweet cherry germplasm characterization. Out of 21 SSR markers, 16 were polymorphic, producing 177 alleles that varied from 4 to 16 alleles (9.35 on average) with a mean heterozygosity value of 0.82 that produced successful amplifications and revealed DNA polymorphisms. Allele size varied from 95 to 290 bp. Cluster analyses showed that the studied sweet cherry genotypes were classified intofive main groups based mainly on their species characteristics and SSR data. In general, our results did not show a clear structuring of genetic variability within the Iranian diffusion area of sweet cherry, so it was not possible to draw any indications on regions of provenance delimitation. The results of this study contribute to a better understanding of sweet cherry genetic variations in Iran, thus making for more efficient programs aimed at preserving biodiversity and

  9. Effects of elevated CO[sub 2] on growth and chloroplast proteins in Prunus avium

    Energy Technology Data Exchange (ETDEWEB)

    Wilkins, D.; Oosten, J.-J. van; Besford, R.T. (Horticulture Research International, Littlehampton, Sussex (United Kingdom))

    1994-01-01

    A study was conducted of the growth response of Prunus avium L. Stella (wild cherry) to elevated CO[sub 2]. The associated changes in photosynthetic machinery of the leaf tissue were characterized. Self-pollinated seedlings and mature cuttings (clones) from the same parent plant of P. avium were grown for two consecutive growing seasons (about 60 days each) in ambient or elevated CO[sub 2] with high or low nutrient supply. The degree of acclimation of leaf biochemistry and growth response to elevated CO[sub 2] depended on the plant material (seedling or mature cutting) and nutrient supply. There was little or no growth response to elevated CO[sub 2] in seedlings or cuttings in the low nutrient supply treatments, whereas in both seasons, there was a strongly positive growth response to elevated CO[sub 2] in seedlings and cuttings in the high nutrient supply regimes, resulting in increases in the root/shoot ratio and in carbon allocation to the roots. In contrast, the protein content and activity of ribulose-1,5-biophosphate carboxylase-oxygenase (Rubisco, EC 4.1.1.39) were down regulated in elevated CO[sub 2]. The loss of Rubisco on an area basis in plants in the elevated CO[sub 2] treatments was compensated for at the canopy level by increased leaf area. The loss of Rubisco protein was accompanied by decreases in the contents of chlorophyll and the thylakoid membrane proteins D[sub 1], D[sub 2] and cytochrome f, which are involved in light harvesting and photo-electron transport. It is concluded that in the medium- to long-term, the initial stimulation of biomass production by elevated CO[sub 2] may be increasingly offset by a lower photosynthetic capacity per unit leaf area in perennial plants. 27 refs., 2 figs., 3 tabs.

  10. Mycobacterium avium Subspecies paratuberculosis Recombinant Proteins Modulate Antimycobacterial Functions of Bovine Macrophages.

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    John P Bannantine

    Full Text Available It has been shown that Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis activates the Mitogen Activated Protein Kinase (MAPK p38 pathway, yet it is unclear which components of M. paratuberculosis are involved in the process. Therefore, a set of 42 M. paratuberculosis recombinant proteins expressed from coding sequences annotated as lipoproteins were screened for their ability to induce IL-10 expression, an indicator of MAPKp38 activation, in bovine monocyte-derived macrophages. A recombinant lipoprotein, designated as MAP3837c, was among a group of 6 proteins that strongly induced IL-10 gene transcription in bovine macrophages, averaging a 3.1-fold increase compared to non-stimulated macrophages. However, a parallel increase in expression of IL-12 and TNF-α was only observed in macrophages exposed to a subset of these 6 proteins. Selected recombinant proteins were further analyzed for their ability to enhance survival of M. avium within bovine macrophages as measured by recovered viable bacteria and nitrite production. All 6 IL-10 inducing MAP recombinant proteins along with M. paratuberculosis cells significantly enhanced phosphorylation of MAPK-p38 in bovine macrophages. Although these proteins are likely not post translationally lipidated in E. coli and thus is a limitation in this study, these results form the foundation of how the protein component of the lipoprotein interacts with the immune system. Collectively, these data reveal M. paratuberculosis proteins that might play a role in MAPK-p38 pathway activation and hence in survival of this organism within bovine macrophages.

  11. Fabrication of a Novel Conductometric Biosensor for Detecting Mycobacterium avium subsp. paratuberculosis Antibodies

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    Steven Bolin

    2008-09-01

    Full Text Available Johne’s disease (JD is one of the most costly bacterial diseases in cattle. In the U.S., economic losses from the disease have been estimated to exceed $1,500,000,000 per year, mainly from the effects of reduced milk production. Current diagnostic tests for JD are laboratory based and many of those tests require specialized equipment and training. Development of rapid and inexpensive diagnostic assays, which are adapted for point-ofcare applications, would aid in the control of JD. In this study, a polyaniline (Pani-based conductometric biosensor, in an immunomigration format, was fabricated for the detection of serum antibody (IgG against the causal organism of JD, Mycobacterium avium subsp. paratuberculosis (MAP. Immobilized Mycobacterium avium purified proteins in the capture membrane were used to detect MAP IgG, previously bound with Pani/anti-bovine IgG* conjugate in the conjugate membrane. After detection, the Pani in the sandwiched captured complex bridges an electrical circuit between the silver electrodes, flanking the capture membrane. The electrical conductance, caused by Pani, was measured as drop in electrical resistance. Testing of the biosensor with known JD positive and negative serum samples demonstrated a significant difference in the mean resistance observed between the groups. This proof-of-concept study demonstrated that a conductometric biosensor could detect MAP IgG in 2 minutes. The biosensor’s speed of detection and the equipment involved would, among other things, support its application towards the various point-ofcare opportunities aimed at JD management and control.

  12. Causation of Crohn’s Disease by Mycobacterium avium Subspecies Paratuberculosis

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    John Hermon-Taylor

    2000-01-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (MAP is a member of the M avium complex (MAC. It differs genetically from other MAC in having 14 to 18 copies of IS900 and a single cassette of DNA involved in the biosynthesis of surface carbohydrate. Unlike other MAC, MAP is a specific cause of chronic inflammation of the intestine in many animal species, including primates. The disease ranges from pluribacillary to paucimicrobial, with chronic granulomatous inflammation like leprosy in humans. MAP infection can persist for years without causing clinical disease. The herd prevalence of MAP infection in Western Europe and North America is reported in the range 21% to 54%. These subclinically infected animals shed MAP in their milk and onto pastures. MAP is more robust than tuberculosis, and the risk that is conveyed to human populations in retail milk and in domestic water supplies is high. MAP is harboured in the ileocolonic mucosa of a proportion of normal people and can be detected in a high proportion of full thickness samples of inflamed Crohn’s disease gut by improved culture systems and IS900 polymerase chain reaction if the correct methods are used. MAP in Crohn’s disease is present in a protease-resistant nonbacillary form, can evade immune recognition and probably causes an immune dysregulation. As with other MAC, MAP is resistant to most standard antituberculous drugs. Treatment of Crohn’s disease with combinations of drugs more active against MAC such as rifabutin and clarithromycin can bring about a profound improvement and, in a few cases, apparent disease eradication. New drugs as well as effective MAP vaccines for animals and humans are needed. The problems caused by MAP constitute a public health issue of tragic proportions for which a range of remedial measures are urgently needed.

  13. Host responses to the pathogen Mycobacterium avium subsp. paratuberculosis and beneficial microbes exhibit host sex specificity.

    Science.gov (United States)

    Karunasena, Enusha; McMahon, K Wyatt; Chang, David; Brashears, Mindy M

    2014-08-01

    Differences between microbial pathogenesis in male and female hosts are well characterized in disease conditions connected to sexual transmission. However, limited biological insight is available on variances attributed to sex specificity in host-microbe interactions, and it is most often a minimized variable outside these transmission events. In this work, we studied two gut microbes-a pathogen, Mycobacterium avium subsp. paratuberculosis, and a probiotic, Lactobacillus animalis NP-51-and the interaction between each agent and the male and female gastrointestinal systems. This trial was conducted in BALB/c mice (n=5 per experimental group and per sex at a given time point), with analysis at four time points over 180 days. Host responses to M.avium subsp. paratuberculosis and L. animalis were sensitive to sex. Cytokines that were significantly different (P ≤ 0.05) betweenthe sexes included interleukin-1α/β (IL-1α/β), IL-17, IL-6, IL-10, IL-12, and gamma interferon (IFN-) and were dependent on experimental conditions. However, granulocyte-macrophage colony-stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF), and IL-13/23 showed no sex specificity. A metabolomics study indicated a 0.5- to 2.0-fold (log2 scale) increase in short-chain fatty acids (butyrate and acetate) in males and greater increases in o-phosphocholine or histidine from female colon tissues; variances distinct to each sex were observed with age or long-term probiotic consumption. Two genera, Staphylococcus and Roseburia, were consistently overrepresented in females compared to males; other species were specific to one sex but fluctuated depending on experimental conditions. The differences observed suggest that male and female gut tissues and microbiota respond to newly introduced microorganisms differently and that gut-associated microorganisms with host immune system responses and metabolic activity are supported by biology distinct to the host sex.

  14. Isolation and Identification of Two Marine-Originated Actinoalloteichus sp. Producing Caerulomycin A%2株海洋来源产浅蓝霉素A的异壁放线菌的分离和鉴定

    Institute of Scientific and Technical Information of China (English)

    纪昌涛; 周潇; 陈奇; 宋永相; 黄洪波; 王博; 鞠建华

    2015-01-01

    利用抗菌及卤虫致死活性模型,从中国南海海底沉积物来源的微生物中筛选到2株放线菌SCSIO WJ01和SCSIO ZJ63,其发酵产物具有较强活性,经16S rRNA基因序列分析这2株放线菌均为异壁放线菌Ac-tinoalloteichus sp.。HPLC-DAD分析显示2株放线菌能产生同一个主要的次级代谢产物,通过正相硅胶柱色谱、反相中压柱色谱、半制备高效液相色谱等手段,从SCSIO WJ01的发酵产物中分离获得了该化合物,运用ESI-MS、1 H及13 C NMR波谱分析鉴定为浅蓝霉素A( Caerulomycin A)。此外,还从SCSIO WJ01的发酵产物中分离鉴定了浅蓝霉素D。%Two marine-originated actinobacteria strains SCSIO WJ01 and SCSIO ZJ63 isolated and screened from sed-iments collected in the bottom of South China Sea,using antibiotic and lethal activities model against brine shrimp ( Artemia salina),their fermentation broths possessed fairly strong activity. These two strains were identified as Acti-noalloteichus sp. through 16S rRNA gene sequence analysis. HPLC-DAD analysis disclosed that strains SCSIO WJ01 and SCSIO ZJ63 could produce the same major secondary metabolite. These fermentation products were isolated from the fermentation extract of strain SCSIO WJ01 by means of ortho-phase silica gel column chromatography and counter-phase mid-pressure column chromatography,and semi-preparative HPLC etc and identified to be caerulomycin A using ESI-MS,1 H and 13 C NMR spectroscopic analyses. In addition,caerulomycin D was also isolated and identified from fermentation product of strain SCSIO WJ01.

  15. Isolation,Identiifcation and Antimicrobial Susceptibility Test to Escherichia coli of Animal Origin in Shanghai City%上海市动物源性大肠杆菌的分离鉴定和药敏试验

    Institute of Scientific and Technical Information of China (English)

    宁昆; 沈莉萍; 王晓旭; 徐锋; 张维谊

    2016-01-01

    In order to investigate the antibiotic resistance of Escherichia coli of animal origin in Shanghai, antimicrobial susceptibility test of 236 E.coli isolates from animal clinics to 14 antimicrobial agents was surveyed. Among them,164 strains were identified as swine origin,72 strains were identified as avian origin. Broth microdilution method according to the recommendation of the CLSI was used. The results showed that both E. coli from swine and avian showed antibiotic resistance to most antimicrobial agents. E.coli strains were highly resistant to sulfisoxazole,sulfamethoxazole,enrofloxacin and spectinomycin,etc. Lower antibiotic resistance rate( ≤ 20%) against amikacin and amoxicillin or clavulanate was showed. In addition,E.coli from swine had a more complex type of drug resistance spectrum than E.coli from avian.%为掌握上海市动物源性大肠杆菌的耐药情况及耐药规律,对2013—2015年在门诊畜禽中分离到的大肠杆菌进行药敏试验。共分离到236株大肠杆菌,其中猪源164株、禽源72株。选用14种抗生素,采用CLSI推荐的微量肉汤稀释法,对大肠杆菌进行药物敏感试验。结果表明,无论是猪源大肠杆菌,还是禽源大肠杆菌,对大多数抗生素的耐药程度均较高。其中,对磺胺异恶唑、复方新诺明、恩诺沙星和大观霉素等抗生素的耐药情况较为严重;对丁胺卡那霉素和阿莫西林/棒酸2种抗生素较为敏感,耐药率低于20%。从耐药谱型来看,猪源大肠杆菌较禽源大肠杆菌具有更加复杂的耐药谱型。

  16. Efficacy of various pasteurization time-temperature conditions in combination with homogenization on inactivation of Mycobacterium avium subsp. paratuberculosis in milk.

    Science.gov (United States)

    Grant, Irene R; Williams, Alan G; Rowe, Michael T; Muir, D Donald

    2005-06-01

    The effect of various pasteurization time-temperature conditions with and without homogenization on the viability of Mycobacterium avium subsp. paratuberculosis was investigated using a pilot-scale commercial high-temperature, short-time (HTST) pasteurizer and raw milk spiked with 10(1) to 10(5) M. avium subsp. paratuberculosis cells/ml. Viable M. avium subsp. paratuberculosis was cultured from 27 (3.3%) of 816 pasteurized milk samples overall, 5 on Herrold's egg yolk medium and 22 by BACTEC culture. Therefore, in 96.7% of samples, M. avium subsp. paratuberculosis had been completely inactivated by HTST pasteurization, alone or in combination with homogenization. Heat treatments incorporating homogenization at 2,500 lb/in2, applied upstream (as a separate process) or in hold (at the start of a holding section), resulted in significantly fewer culture-positive samples than pasteurization treatments without homogenization (P homogenization was estimated to be 4.0 to 5.2 log10. The impact of homogenization on clump size distribution in M. avium subsp. paratuberculosis broth suspensions was subsequently assessed using a Mastersizer X spectrometer. These experiments demonstrated that large clumps of M. avium subsp. paratuberculosis cells were reduced to single-cell or "miniclump" status by homogenization at 2,500 lb/in2. Consequently, when HTST pasteurization was being applied to homogenized milk, the M. avium subsp. paratuberculosis cells would have been present as predominantly declumped cells, which may possibly explain the greater inactivation achieved by the combination of pasteurization and homogenization.

  17. Characterization of the intracellular survival of Mycobacterium avium ssp. paratuberculosis: phagosomal pH and fusogenicity in J774 macrophages compared with other mycobacteria.

    Science.gov (United States)

    Kuehnel, M P; Goethe, R; Habermann, A; Mueller, E; Rohde, M; Griffiths, G; Valentin-Weigand, P

    2001-08-01

    The phagosomes containing viable pathogenic mycobacteria, such as Mycobacterium (M.) tuberculosis and Mycobacterium avium ssp. avium (M. avium), are known to be limited in their ability to both acidify and fuse with late (but not early) endocytic organelles. Here, we analysed the pH and fusogenicity of phagosomes containing M. avium ssp. paratuberculosis (M. ptb), the causative agent of paratuberculosis in ruminants. Using the murine J774 macrophage cell line, we compared viable and heat-killed M. ptb and, in addition, viable or dead M. avium, as well as two non-pathogenic mycobacteria, Mycobacterium smegmatis and Mycobacterium gordonae. Electron microscopic analysis revealed that M. ptb persisted intracellularly in phagosomes for up to 15 days. The phagosomes containing live M. ptb and M. avium were significantly reduced in their ability to acquire some markers for the endocytic pathway, such as internalized calcein, BSA-gold or the membrane protein Lamp 2. However, they were almost completely accessible to 70 kDa fluorescein isothiocyanate (FITC)-dextran and Lamp 1. Overall, the phagosomes containing dead pathogenic mycobacteria behaved similarly to the ones containing live non-pathogenic mycobacteria in all experiments. Using FITC-dextran in a novel fluorescence-activated cell sorting (FACS)-based method, we could also show that the bulk of endocytic compartments, including phagosomes, were only very mildly acidified to approximately pH 6.3 over at least 72 h in J774 cells infected with live M. ptb and M. avium. In contrast, J774 cells treated with heat-killed M. ptb or BSA-coated latex beads showed substantial acidification of the phagosome/endocytic compartments to a pH value of approximately 5.2. After infection with M. smegmatis and M. gordonae, acidification was initially (1-5 h after infection) inhibited, but increased after longer infection to levels similar to those with dead mycobacteria.

  18. Identification of (+)-erythro-mefloquine as an active enantiomer with greater efficacy than mefloquine against Mycobacterium avium infection in mice.

    Science.gov (United States)

    Bermudez, Luiz E; Inderlied, Clark B; Kolonoski, Peter; Chee, Christopher B; Aralar, Priscilla; Petrofsky, Mary; Parman, Toufan; Green, Carol E; Lewin, Anita H; Ellis, William Y; Young, Lowell S

    2012-08-01

    Infection caused by Mycobacterium avium is common in AIDS patients who do not receive treatment with highly active antiretroviral therapy (HAART) or who develop resistance to anti-HIV therapy. Mefloquine, a racemic mixture used for malaria prophylaxis and treatment, is bactericidal against M. avium in mice. MICs of (+)-erythro-, (-)-erythro-, (+)-threo-, and (-)-threo-mefloquine were 32 μg/ml, 32 μg/ml, 64 μg/ml, and 64 μg/ml, respectively. The postantibiotic effect for (+)-erythro-mefloquine was 36 h (MIC) and 41 h for a concentration of 4× MIC. The mefloquine postantibiotic effect was 25 h (MIC and 4× MIC). After baseline infection was established (7 days), the (+)- and (-)-isomers of the diastereomeric threo- and erythro-α-(2-piperidyl)-2,8-bis(trifluoromethyl)-4-quinolinemethanol were individually used to orally treat C57BL/6 bg(+)/bg(+) beige mice that were infected intravenously with M. avium. Mice were also treated with commercial mefloquine and diluent as controls. After 4 weeks of treatment, the mice were harvested, and the number of bacteria in spleen and liver was determined. Mice receiving (+)- or (-)-threo-mefloquine or (-)-erythro-mefloquine had numbers of bacterial load in tissues similar to those of untreated control mice at 4 weeks. Commercial mefloquine had a bactericidal effect. However, mice given the (+)-erythro-enantiomer for 4 weeks had a significantly greater reduction of bacterial load than those given mefloquine. Thus, (+)-erythro-mefloquine is the active enantiomer of mefloquine against M. avium and perhaps other mycobacteria.

  19. The clinical efficacy of a clarithromycin-based regimen for Mycobacterium avium complex disease: A nationwide post-marketing study.

    Science.gov (United States)

    Kadota, Jun-Ichi; Kurashima, Atsuyuki; Suzuki, Katsuhiro

    2017-02-26

    The revised 2007 American Thoracic Society/Infectious Diseases Society of America statement recommend clarithromycin-based combination therapy for treatment of Mycobacterium avium complex lung disease and stipulates approximately 1 year of continuous treatment after bacilli negative conversion. However, supporting data are insufficient. Our objective was to obtain data on the clinical outcome of clarithromycin-based daily regimens by conducting a nationwide retrospective post-marketing study of M.avium complex lung disease. In accordance with the Japanese guidelines, patients were enrolled in this survey according to their chest radiographic findings and microbiologic test results. They were treated with a multidrug regimen including clarithromycin, rifampicin, and ethambutol (clarithromycin-based regimen) until bacilli negative conversion, and the treatment was continued for approximately 1 year after the initial conversion. Data were collected before administration, at the time of bacilli negative conversion, at the end of treatment, and at 6 months after the end of treatment. Of the 466 subjects enrolled in the study, 271 patients who received clarithromycin at 800 mg/day underwent evaluation for M.avium complex disease. The final bacilli negative conversion rate in those patients was 94.7%. The bacteriological relapse rate was 5.0% (5/100 patients). Bacteriological relapse was noted in patients treated for less than 15 months after conversion. No life-threatening or serious adverse drug reactions were observed. This study demonstrated that a clarithromycin-based daily regimen can yield a high bacteriological conversion rate in M.avium complex disease. After conversion, treatment for less than 15 months might be insufficient to prevent bacteriological relapse.

  20. Response to Switch from Intermittent Therapy to Daily Therapy for Refractory Nodular Bronchiectatic Mycobacterium avium Complex Lung Disease

    OpenAIRE

    Koh, Won-Jung; Jeong, Byeong-Ho; Jeon, Kyeongman; Park, Hye Yun; Kim, Su-Young; Huh, Hee Jae; Ki, Chang-Seok; Lee, Nam Yong; Shin, Sung Jae; Daley, Charles L.

    2015-01-01

    Intermittent three-times-weekly antibiotic therapy is recommended for the initial treatment of patients with noncavitary nodular bronchiectatic Mycobacterium avium complex lung disease. Although some experts recommend switching from intermittent to daily therapy for patients whose sputum has persistent positive cultures after intermittent therapy, the clinical efficacy of these modifications is unknown. Of 20 patients whose sputum had persistent positive cultures after 12 months of intermitte...

  1. Mycobacterium avium MAV2052 protein induces apoptosis in murine macrophage cells through Toll-like receptor 4.

    Science.gov (United States)

    Lee, Kang-In; Choi, Han-Gyu; Son, Yeo-Jin; Whang, Jake; Kim, Kwangwook; Jeon, Heat Sal; Park, Hye-Soo; Back, Yong Woo; Choi, Seunga; Kim, Seong-Woo; Choi, Chul Hee; Kim, Hwa-Jung

    2016-04-01

    Mycobacterium avium and its sonic extracts induce apoptosis in macrophages. However, little is known about the M. avium components regulating macrophage apoptosis. In this study, using multidimensional fractionation, we identified MAV2052 protein, which induced macrophage apoptosis in M. avium culture filtrates. The recombinant MAV2052 induced macrophage apoptosis in a caspase-dependent manner. The loss of mitochondrial transmembrane potential (ΔΨm), mitochondrial translocation of Bax, and release of cytochrome c from mitochondria were observed in macrophages treated with MAV2052. Further, reactive oxygen species (ROS) production was required for the apoptosis induced by MAV2052. In addition, ROS and mitogen-activated protein kinases were involved in MAV2052-mediated TNF-α and IL-6 production. ROS-mediated activation of apoptosis signal-regulating kinase 1 (ASK1)-JNK pathway was a major signaling pathway for MAV2052-induced apoptosis. Moreover, MAV2052 bound to Toll-like receptor (TLR) 4 molecule and MAV2052-induced ROS production, ΔΨm loss, and apoptosis were all significantly reduced in TLR4(-/-) macrophages. Altogether, our results suggest that MAV2052 induces apoptotic cell death through TLR4 dependent ROS production and JNK pathway in murine macrophages.

  2. Virulence-related Mycobacterium avium subsp hominissuis MAV_2928 gene is associated with vacuole remodeling in macrophages

    Directory of Open Access Journals (Sweden)

    Vogt Steven

    2010-04-01

    Full Text Available Abstract Background Mycobacterium avium subsp hominissuis (previously Mycobacterium avium subsp avium is an environmental organism associated with opportunistic infections in humans. Mycobacterium hominissuis infects and replicates within mononuclear phagocytes. Previous study characterized an attenuated mutant in which the PPE gene (MAV_2928 homologous to Rv1787 was inactivated. This mutant, in contrast to the wild-type bacterium, was shown both to have impaired the ability to replicate within macrophages and to have prevented phagosome/lysosome fusion. Results MAV_2928 gene is primarily upregulated upon phagocytosis. The transcriptional profile of macrophages infected with the wild-type bacterium and the mutant were examined using DNA microarray, which showed that the two bacteria interact uniquely with mononuclear phagocytes. Based on the results, it was hypothesized that the phagosome environment and vacuole membrane of the wild-type bacterium might differ from the mutant. Wild-type bacterium phagosomes expressed a number of proteins different from those infected with the mutant. Proteins on the phagosomes were confirmed by fluorescence microscopy and Western blot. The environment in the phagosome of macrophages infected with the mutant differed from the environment of vacuoles with M. hominissuis wild-type in the concentration of zinc, manganese, calcium and potassium. Conclusion The results suggest that the MAV_2928 gene/operon might participate in the establishment of bacterial intracellular environment in macrophages.

  3. Residual effects of Mycobacterium avium infection on susceptibility of sheep to copper toxicity and efficacy of treatment with tetrathiomolybdate.

    Science.gov (United States)

    Suttle, N F

    2012-07-07

    Posthaemolytic copper poisoning (post-HCP) in one of six, one-year-old, uninfected sheep (group O) on a Mycobacterium avium experiment prompted an evaluation of copper status and hepatotoxicity in 17 surviving cohorts. Group O had higher mean plasma Cu and δ-glutamyl transferase (GGT) activity and more variable bile acid (BA) concentrations and glutamate dehydrogenase activities (GDH) than two groups infected with M avium soon after birth. Ammonium tetrathiomolybdate (TTM; 3 x 1.7 mg/kg LW) was given subcutaneously over seven days and the pelleted, complete diet replaced by hay, low in copper. Plasma BA immediately declined and was followed by GDH, but erythrocyte superoxide dismutase activity (ESOD) became severely inhibited and took 18 days to recover. Plasma BA and GDH rose sharply after 18 days in uninfected sheep and they became hypercupraemic. TTM treatment was repeated from day 42 and had removed all group differences by day 110 but only after further inhibition of ESOD. M avium infections probably lessened the severity of pre-HCP by reducing copper retention but may predispose grazing livestock to hypocupraemia. The capacity of TTM to reduce liver Cu has probably been overestimated and side effects on cuproenzyme activity underestimated.

  4. Antibiotic resistances in Listeria monocytogenes and Salmonella enterica isolated from foods with animal origin Resistencias a antibióticos en Listeria monocytogenes y Salmonella enterica aislados de alimentos de origen animal

    Directory of Open Access Journals (Sweden)

    Baltasar Balsalobre Hernández

    2004-12-01

    Full Text Available Extensive use of antibiotics in both human and animal health and in cattle production has generated resistant microorganisms to common antibiotics. Resistances spread caused by human and animal therapeutic is well known, but we know poorly frecuency of resistant bacteria in foods with animal origin and destinated to human consumers. In this paper, sensitivity to nineteen antibiotics was investigated in Listeria monocytogenes and Salmonella enterica strains isolated from foods with animal origin, including fresh meat, hamburgers, fresh sausages, boiled ham and new-laid chicken eggs. The plate diffusion method of Bauer-Kirby was used.Listeria monocytogenes strains showed a very high sensitivity to all antibiotics checked, with the exception of one strain tetracycline resistant. In contrast, Salmonella enterica showed a high frecuency of resistances, in special to tetracycline, streptomycin, nalidixic acid, ticarcillin, ampicillin and chloramphenicol. Moreover, multi-resistance was a common phenomenon. Twenty percent of S. enterica strains were resistant to four or more antibiotics. Frecuency of resistances was higher in 4,5,12:i:-, Hadar, Typhimurium and Virchow serotypes.In conclusion, Salmonella enterica strains isolated from foods with animal origin and destinated to human consumers are usually resistant to several antibiotics. The significance of this observation and its potential health risk must be investigated.El uso extensivo de antibióticos para la salud humana y animal así como para mejorar la producción ganadera ha generado un gran número de cepas microbianas resistentes a antibióticos de uso común. Es bien conocida la difusión de resistencias a través de la terapéutica humana y animal, pero desconocemos en qué medida los alimentos de origen animal destinados a consumo humano son portadores de resistencias.En este trabajo, se investigó la sensibilidad a diecinueve antibióticos de cepas de Listeria monocytogenes y Salmonella

  5. Multiple isolates from Aids patients: aspects of an analysis by a genotypic marker and antimicrobial susceptibilities variations

    Directory of Open Access Journals (Sweden)

    Maria Helena Féres Saad

    2000-10-01

    Full Text Available Twenty-one Mycobacterium avium multisolates, from ten human immunodeficiency virus-infected patients, were typed by restriction fragment length polymorphism using as marker the IS1245 and characterized by minimum inhibitory concentration for nine different antibiotics. Two out of four patients harboring multisolates with different fingerprint profile, were therefore considered as having a polyclonal infection, since their isolates were taken from sterile site. This result confirms that polyclonal infection caused by M. avium occurs with a nonnegligenciable frequency. Analyzing the multisolates susceptibility profile of each patient it was observed that most of them were infected with strains having appreciably different antimicrobial susceptibility patterns, no matter what the genotypic pattern of the strains was. These results have strong implication for the treatment of the patients.

  6. Original pedagogues

    DEFF Research Database (Denmark)

    Schmidt, Christina Haandbæk

    Original pedagogues Distention between competences and originality By Christina Haandbæk Schmidt, ph. d. student Aarhus University, Denmark This presentation concerns a Ph.D. project (Sept. 2012 –Sept. 2015) about pedagogues in day care facilities and their struggles to develop and retain...... of pedagogues and in everyday life in daycare facilities. The competence term includes at least two discourses of interest; a pedagogical competence discourse and a political jurisdiction discourse which forms a distention between authenticity and competence. In order that pedagogues may regain their autonomy I...

  7. Apa antigen of Mycobacterium avium subsp. paratuberculosis as a target for species-specific immunodetection of the bacteria in infected tissues of cattle with paratuberculosis.

    Science.gov (United States)

    Souza, Giliane S; Rodrigues, Ana Bárbara F; Gioffré, Andrea; Romano, Maria I; Carvalho, Eulógio C Q; Ventura, Thatiana L B; Lasunskaia, Elena B

    2011-09-15

    Comparative genomics of Mycobacterium spp. have revealed conservative genes and respective proteins differently expressed in mycobacteria that could be used as targets for the species-specific immunodiagnostics. The alanine and proline-rich antigen Apa is a mycobacterial protein that present significant variability in primary sequence length and composition between members of M. avium and M. tuberculosis complexes. In this study, the recombinant Apa protein encoded by the MAP1569/ModD gene of M. avium subsp. paratuberculosis (Map) was used to generate a panel of monoclonal antibodies which were shown to recognize the most important veterinary pathogens of the M. avium complex, specifically Map and M. avium subsp. hominissuis, and which did not cross-react with M. bovis or M. tuberculosis. The produced antibodies were demonstrated to be a useful tool for the species-specific immunofluorescence or immunohistochemical detection of Map in experimentally infected cell cultures or intestinal tissues from cattle with bovine paratuberculosis and, additionally, they may be employed for the discrimination of pathogenic M. avium subspecies via Western blotting.

  8. Original Copies

    DEFF Research Database (Denmark)

    Sørensen, Tim Flohr

    2013-01-01

    of similarity by looking at artefactual similarity as the results of prototyping and as a production of simulacra. In this light, the concept of copying turns out to be more than simply a matter of trying to imitate an exotic or prestigious original, and it fundamentally raises the question how different a copy...

  9. Occurrence of Mycobacterium avium subspecies paratuberculosis across host species and European countries with evidence for transmission between wildlife and domestic ruminants

    Directory of Open Access Journals (Sweden)

    May Linda

    2009-10-01

    Full Text Available Abstract Background Mycobacterium avium subspecies paratuberculosis (Map causes an infectious chronic enteritis (paratuberculosis or Johne's disease principally of ruminants. The epidemiology of Map is poorly understood, particularly with respect to the role of wildlife reservoirs and the controversial issue of zoonotic potential (Crohn's disease. Genotypic discrimination of Map isolates is pivotal to descriptive epidemiology and resolving these issues. This study was undertaken to determine the genetic diversity of Map, enhance our understanding of the host range and distribution and assess the potential for interspecies transmission. Results 164 Map isolates from seven European countries representing 19 different host species were genotyped by standardized IS900 - restriction fragment length polymorphism (IS900-RFLP, pulsed-field gel electrophoresis (PFGE, amplified fragment length polymorphisms (AFLP and mycobacterial interspersed repeat unit-variable number tandem repeat (MIRU-VNTR analyses. Six PstI and 17 BstEII IS900-RFLP, 31 multiplex [SnaBI-SpeI] PFGE profiles and 23 MIRU-VNTR profiles were detected. AFLP gave insufficient discrimination of isolates for meaningful genetic analysis. Point estimates for Simpson's index of diversity calculated for the individual typing techniques were in the range of 0.636 to 0.664 but a combination of all three methods increased the discriminating power to 0.879, sufficient for investigating transmission dynamics. Two predominant strain types were detected across Europe with all three typing techniques. Evidence for interspecies transmission between wildlife and domestic ruminants on the same property was demonstrated in four cases, between wildlife species on the same property in two cases and between different species of domestic livestock on one property. Conclusion The results of this study showed that it is necessary to use multiple genotyping techniques targeting different sources of genetic

  10. MEDIAMUSIC ORIGINS

    Directory of Open Access Journals (Sweden)

    Chernyshov Alexander V.

    2012-12-01

    Full Text Available The article investigates the origins of music in electronic regular broadcasting, which conditions have appeared in the XIX century. They stand out in the "telephone concerts", "phonograph concerts" and the proto-sound films (T.Edison, W.Dickson, Ch.Pathé, O.Messter. In the early twentieth century, a clear prototype of mediamusic playing by the music of "silent" films, which has been divided on the on-screen and offscreen sound layers, the method of compilation, the basics of synchronization between musical sound and off-music montage-structures. In addition, the origins of music of electronic mass media can be regarded as attempts to understand the "musical" noise features, which subsequently materialize in the phenomenon of "noisemusic" of media audio-score (L.Russolo, Ars.Avraamov, D.Vertov, W.Ruttmann, N.Voinov, P.Schaeffer. Are considered Russian, European and North American experiences.

  11. Mitochondrial origins.

    OpenAIRE

    Yang, D.; Oyaizu, Y; Oyaizu, H; Olsen, G J; Woese, C R

    1985-01-01

    The 16S ribosomal RNA sequences from Agrobacterium tumefaciens and Pseudomonas testosteroni have been determined to further delimit the origin of the endosymbiont that gave rise to the mitochondrion. These two prokaryotes represent the alpha and beta subdivisions, respectively, of the so-called purple bacteria. The endosymbiont that gave rise to the mitochondrion belonged to the alpha subdivision, a group that also contains the rhizobacteria, the agrobacteria, and the rickettsias--all prokary...

  12. Non-tuberculous mycobacteria I: one year clinical isolates identification in Tertiary Hospital Aids Reference Center, Rio de Janeiro, Brazil, in pre highly active antiretroviral therapy era

    Directory of Open Access Journals (Sweden)

    Ferreira Rosa Maria Carvalho

    2002-01-01

    Full Text Available The aim of this study was to determine the prevalence of non-tuberculous mycobacteria (NTM isolates at University Hospital, Reference Center for Aids in Rio de Janeiro, Brazil, during one year. We used standard biochemical tests for species identification and IS1245 PCR amplification was applied as a Mycobacterium avium specific identification marker. Four hundred and four specimens from 233 patients yielded acid-fast bacilli growth. M. tuberculosis was identified in 85% of the patients and NTM in 15%. NTM disseminated infection was a common event correlated with human immunodeficiency virus (HIV infected patients and only in HIV negative patients the source of NTM was non sterile site. M. avium complex (MAC was biochemically identified in 57.8% (49/83 of NTM isolates, most of them from sterile sites (75.5%, and in 94% (46/49 the IS 1245 marker specific for M. avium was present. Twenty NTM strains showed a MAC biochemical pattern with the exception of a urease-positive (99% of MAC are urease-negative, however IS1245 was detected in 96% of the strains leading to their identification as M. avium. In this group differences in NTM source was not significant. The second most frequently isolated NTM was identified as M. scrofulaceum (7.2%, followed by M. terrae (3.6%, M. gordonae (2.4%, M. chelonae (1.2%, M. fortuitum (1.2% and one strain which could not be identified. All were IS1245 negative except for one strain identified as M. scrofulaceum. It is interesting to note that non-sterile sites were the major source of these isolates (92.8%. Our finding indicated that M. avium is still the major atypical species among in the MAC isolates recovered from Brazilian Aids patients without highty active antiretroviral therapy schema. Some discrepancies were seen between the identification methods and further investigations must be done to better characterize NTM isolates using other phenotypic and genotypic methods.

  13. Use of conventional and real-time polymerase chain reaction for confirmation of Mycobacterium avium subsp. paratuberculosis in a broth-based culture system ESP II.

    Science.gov (United States)

    Kim, Sung G; Kim, Eun H; Lafferty, Caroline J; Miller, Loretta J; Koo, Hye J; Stehman, Susan M; Shin, Sang J

    2004-09-01

    The ESP II Culture System (ESP II), a broth-based culture system, has been modified and optimized for culturing Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) in animal feces since 2000. Conventional and real-time polymerase chain reaction (PCR) assays based on the IS900 sequence were performed as confirmatory tests for M. paratuberculosis in ESP II liquid culture medium. There were no differences between test results of conventional and real-time PCR assays. During the 5-week incubation period, if acid-fast bacilli (AFB) were detected in ESP culture-positive samples, IS900 PCR assays were performed to confirm whether those AFB were M. paratuberculosis. At the end of the 5-week incubation, AF staining was performed on all ESP II-negative cultures to screen any false-negative cultures; IS900 PCR assays were performed on AFB-positive cultures. During a period of 1 year, of a total of 18,499 ESP II cultures, 2,814 (15.2%) PCR confirmation assays were performed. Of those, 2,259 (80%) were both ESP and PCR positive; 104 (4%) were ESP positive and PCR negative; 423 (15%) were ESP negative and PCR positive; 28 (1%) were both ESP and PCR negative. The AF-staining step after the 5-week incubation produced 423 (15%) more PCR-positive cultures. Of a total of 2,814 AFB-positive cultures, 132 (5%) were not confirmed as M. paratuberculosis. Further studies are needed for speciation of non-M. paratuberculosis isolates.

  14. A robust method for bacterial lysis and DNA purification to be used with real-time PCR for detection of Mycobacterium avium subsp. paratuberculosis in milk.

    Science.gov (United States)

    Herthnek, David; Nielsen, Søren Saxmose; Lindberg, Ann; Bölske, Göran

    2008-10-01

    A possible mode of transmission for the ruminant pathogen Mycobacterium avium subsp. paratuberculosis (MAP) from cattle to humans is via milk and dairy products. Although controversially, MAP has been suggested as the causative agent of Crohn's disease and its presence in consumers' milk might be of concern. A method to detect MAP in milk with real-time PCR was developed for screening of bulk tank milk. Pellet and cream fractions of milk were pooled and subjected to enzymatic digestion and mechanical disruption and the DNA was extracted by automated magnetic bead separation. The analytical sensitivity was assessed to 100 organisms per ml milk (corresponding to 1-10 CFU per ml) for samples of 10 ml. The method was applied in a study of 56 dairy herds to compare PCR of farm bulk tank milk to culture of environmental faecal samples for detection of MAP in the herds. In this study, 68% of the herds were positive by environmental culture, while 30% were positive by milk PCR. Results indicate that although MAP may be shed into milk or transferred to milk by faecal contamination, it will probably occur in low numbers in the bulk tank milk due to dilution as well as general milking hygiene measures. The concentration of MAP can therefore be assumed to often fall below the detection limit. Thus, PCR detection of MAP in milk would be more useful for control of MAP presence in milk, in order to avoid transfer to humans, than for herd prevalence testing. It could also be of value in assessing human exposure to MAP via milk consumption. Quantification results also suggest that the level of MAP in the bulk tank milk of the studied Danish dairy herds was low, despite environmental isolation of MAP from the herds.

  15. PCR Inhibition of a Quantitative PCR for Detection of Mycobacterium avium Subspecies Paratuberculosis DNA in Feces: Diagnostic Implications and Potential Solutions.

    Science.gov (United States)

    Acharya, Kamal R; Dhand, Navneet K; Whittington, Richard J; Plain, Karren M

    2017-01-01

    Molecular tests such as polymerase chain reaction (PCR) are increasingly being applied for the diagnosis of Johne's disease, a chronic intestinal infection of ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP). Feces, as the primary test sample, presents challenges in terms of effective DNA isolation, with potential for PCR inhibition and ultimately for reduced analytical and diagnostic sensitivity. However, limited evidence is available regarding the magnitude and diagnostic implications of PCR inhibition for the detection of MAP in feces. This study aimed to investigate the presence and diagnostic implications of PCR inhibition in a quantitative PCR assay for MAP (High-throughput Johne's test) to investigate the characteristics of samples prone to inhibition and to identify measures that can be taken to overcome this. In a study of fecal samples derived from a high prevalence, endemically infected cattle herd, 19.94% of fecal DNA extracts showed some evidence of inhibition. Relief of inhibition by a five-fold dilution of the DNA extract led to an average increase in quantification of DNA by 3.3-fold that consequently increased test sensitivity of the qPCR from 55 to 80% compared to fecal culture. DNA extracts with higher DNA and protein content had 19.33 and 10.94 times higher odds of showing inhibition, respectively. The results suggest that the current test protocol is sensitive for herd level diagnosis of Johne's disease but that test sensitivity and individual level diagnosis could be enhanced by relief of PCR inhibition, achieved by five-fold dilution of the DNA extract. Furthermore, qualitative and quantitative parameters derived from absorbance measures of DNA extracts could be useful for prediction of inhibitory fecal samples.

  16. PCR Inhibition of a Quantitative PCR for Detection of Mycobacterium avium Subspecies Paratuberculosis DNA in Feces: Diagnostic Implications and Potential Solutions

    Science.gov (United States)

    Acharya, Kamal R.; Dhand, Navneet K.; Whittington, Richard J.; Plain, Karren M.

    2017-01-01

    Molecular tests such as polymerase chain reaction (PCR) are increasingly being applied for the diagnosis of Johne’s disease, a chronic intestinal infection of ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP). Feces, as the primary test sample, presents challenges in terms of effective DNA isolation, with potential for PCR inhibition and ultimately for reduced analytical and diagnostic sensitivity. However, limited evidence is available regarding the magnitude and diagnostic implications of PCR inhibition for the detection of MAP in feces. This study aimed to investigate the presence and diagnostic implications of PCR inhibition in a quantitative PCR assay for MAP (High-throughput Johne’s test) to investigate the characteristics of samples prone to inhibition and to identify measures that can be taken to overcome this. In a study of fecal samples derived from a high prevalence, endemically infected cattle herd, 19.94% of fecal DNA extracts showed some evidence of inhibition. Relief of inhibition by a five-fold dilution of the DNA extract led to an average increase in quantification of DNA by 3.3-fold that consequently increased test sensitivity of the qPCR from 55 to 80% compared to fecal culture. DNA extracts with higher DNA and protein content had 19.33 and 10.94 times higher odds of showing inhibition, respectively. The results suggest that the current test protocol is sensitive for herd level diagnosis of Johne’s disease but that test sensitivity and individual level diagnosis could be enhanced by relief of PCR inhibition, achieved by five-fold dilution of the DNA extract. Furthermore, qualitative and quantitative parameters derived from absorbance measures of DNA extracts could be useful for prediction of inhibitory fecal samples. PMID:28210245

  17. Detection of Mycobacterium avium subspecies paratuberculosis specific IS900 insertion sequences in bulk-tank milk samples obtained from different regions throughout Switzerland

    Directory of Open Access Journals (Sweden)

    Stephan Roger

    2002-06-01

    Full Text Available Abstract Background Since Mycobacterium avium subspecies paratuberculosis (MAP was isolated from intestinal tissue of a human patient suffering Crohn's disease, a controversial discussion exists whether MAP have a role in the etiology of Crohn's disease or not. Raw milk may be a potential vehicle for the transmission of MAP to human population. In a previous paper, we have demonstrated that MAP are found in raw milk samples obtained from a defined region in Switzerland. The aim of this work is to collect data about the prevalence of MAP specific IS900 insertion sequence in bulk-tank milk samples in different regions of Switzerland. Furthermore, we examined eventual correlation between the presence of MAP and the somatic cell counts, the total colony counts and the presence of Enterobacteriaceae. Results 273 (19.7% of the 1384 examined bulk-tank milk samples tested IS900 PCR-positive. The prevalence, however, in the different regions of Switzerland shows significant differences and ranged from 1.7% to 49.2%. Furthermore, there were no statistically significant (p >> 0.05 differences between the somatic cell counts and the total colony counts of PCR-positive and PCR-negative milk samples. Enterobacteriaceae occur as often in IS900 PCR-positive as in PCR-negative milk samples. Conclusion This is the first study, which investigates the prevalence of MAP in bulk-tank milk samples all over Switzerland and infers the herd-level prevalence of MAP infection in dairy herds. The prevalence of 19.7% IS900 PCR-positive bulk-milk samples shows a wide distribution of subclinical MAP-infections in dairy stock in Switzerland. MAP can therefore often be transmitted to humans by raw milk consumption.

  18. High Level Aminoglycoside Resistance and Distribution of Aminoglycoside Resistant Genes among Clinical Isolates of Enterococcus Species in Chennai, India

    Directory of Open Access Journals (Sweden)

    Elango Padmasini

    2014-01-01

    Full Text Available Enterococci are nosocomial pathogen with multiple-drug resistance by intrinsic and extrinsic mechanisms. Aminoglycosides along with cell wall inhibitors are given clinically for treating enterococcal infections. 178 enterococcal isolates were analyzed in this study. E. faecalis is identified to be the predominant Enterococcus species, along with E. faecium, E. avium, E. hirae, E. durans, E. dispar and E. gallinarum. High level aminoglycoside resistance (HLAR by MIC for gentamicin (GM, streptomycin (SM and both (GM + SM antibiotics was found to be 42.7%, 29.8%, and 21.9%, respectively. Detection of aminoglycoside modifying enzyme encoding genes (AME in enterococci was identified by multiplex PCR for aac(6′-Ie-aph(2′′-Ia; aph(2′′-Ib; aph(2′′-Ic; aph(2′′-Id and aph(3′-IIIa genes. 38.2% isolates carried aac(6′-Ie-aph(2′′-Ia gene and 40.4% isolates carried aph(3′-IIIa gene. aph(2′′-Ib; aph(2′′-Ic; aph(2′′-Id were not detected among our study isolates. aac(6′-Ie-aph(2′′-Ia and aph(3′-IIIa genes were also observed in HLAR E. durans, E. avium, E. hirae, and E. gallinarum isolates. This indicates that high level aminoglycoside resistance genes are widely disseminated among isolates of enterococci from Chennai.

  19. High level aminoglycoside resistance and distribution of aminoglycoside resistant genes among clinical isolates of Enterococcus species in Chennai, India.

    Science.gov (United States)

    Padmasini, Elango; Padmaraj, R; Ramesh, S Srivani

    2014-01-01

    Enterococci are nosocomial pathogen with multiple-drug resistance by intrinsic and extrinsic mechanisms. Aminoglycosides along with cell wall inhibitors are given clinically for treating enterococcal infections. 178 enterococcal isolates were analyzed in this study. E. faecalis is identified to be the predominant Enterococcus species, along with E. faecium, E. avium, E. hirae, E. durans, E. dispar and E. gallinarum. High level aminoglycoside resistance (HLAR) by MIC for gentamicin (GM), streptomycin (SM) and both (GM + SM) antibiotics was found to be 42.7%, 29.8%, and 21.9%, respectively. Detection of aminoglycoside modifying enzyme encoding genes (AME) in enterococci was identified by multiplex PCR for aac(6')-Ie-aph(2'')-Ia; aph(2'')-Ib; aph(2'')-Ic; aph(2'')-Id and aph(3')-IIIa genes. 38.2% isolates carried aac(6')-Ie-aph(2'')-Ia gene and 40.4% isolates carried aph(3')-IIIa gene. aph(2'')-Ib; aph(2'')-Ic; aph(2'')-Id were not detected among our study isolates. aac(6')-Ie-aph(2'')-Ia and aph(3')-IIIa genes were also observed in HLAR E. durans, E. avium, E. hirae, and E. gallinarum isolates. This indicates that high level aminoglycoside resistance genes are widely disseminated among isolates of enterococci from Chennai.

  20. Isolation and identification of porcine origin avian paramyxovirus serotype 1 in Guangxi%广西猪源禽Ⅰ型副粘病毒的分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    杨荣; 何奇松; 伍和明; 冯淑萍; 付薇; 徐贤坤; 蒋家霞; 孙翔翔; 熊毅

    2012-01-01

    [Objective]The present experiment was conducted to further understand the epidemic features, toxicity levels, and genotypes of porcine origin avian paramyxovirus serotype 1 in Guangxi in order to provide the relevant data for carrying out future researches on porcine paramyxovirus, and to provide references for the prevention and control of the disease. [Method]The vims was isolated from suspected samples of pigs using chrcken embryos inoculation. The characteristics of the virus were determined with hemagglutirtalion (HA), hemagglutination-inhibition (HI) test, mean death time of chicken embryos (MDT), and intracerebral phthngenicily index (ICPI). The virus strain was identified and F gene fragments were amplified using RT-PCR and sequenced. [Result]The pig source avian paramyxovirus serotype I isolate was isolated successfully from suspected samples of pigs. Its titer of HA and HI was 26 and chick embryo minimal lethal dose was 10-8/0.l mL. The MDT and ICPI of this isolate were 60 h and 1.45, respectively. Amino acid composition was R-R-Q-R-R-F of F gene 112-117 splitting sites, which was in accordance with those of NDV standard virulent strain HER33 and F48E9. The nucleotide homulogues were 86.5 and 85.7%, respectively, and the amino acid homologues were 90.1 and 90.8% after comparing the F gene of strain with standard virulent strain F48E9 and moderate virulence strain Beaudette C. [Conclusion]The isolate was virulent strain belonging to A PMV-1 gene type II and traditional strains. It signified that hosts of APMV-1 were spreading and expanding constantly in Guangxi area.%[目的]了解广西猪源禽Ⅰ型副粘病毒的流行特征、毒力强弱及基因类型,为今后开展猪源副粘病毒的相关研究提供参考,也为有效防控副粘病毒感染奠定基础.[方法]采集疑似发病猪组织病料,通过鸡胚接种传代分离病毒后,分别进行血凝(HA)与血凝抑制(HI)试验、平均死亡时间(MDT)和脑内接种致死指数(ICPT)测

  1. TGF-β-mediated sustained ERK1/2 activity promotes the inhibition of intracellular growth of Mycobacterium avium in epithelioid cells surrogates.

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    Carolina L'Abbate

    Full Text Available Transforming growth factor beta (TGF-β has been implicated in the pathogenesis of several diseases including infection with intracellular pathogens such as the Mycobacterium avium complex. Infection of macrophages with M. avium induces TGF-β production and neutralization of this cytokine has been associated with decreased intracellular bacterial growth. We have previously demonstrated that epithelioid cell surrogates (ECs derived from primary murine peritoneal macrophages through a process of differentiation induced by IL-4 overlap several features of epithelioid cells found in granulomas. In contrast to undifferentiated macrophages, ECs produce larger amounts of TGF-β and inhibit the intracellular growth of M. avium. Here we asked whether the levels of TGF-β produced by ECs are sufficient to induce a self-sustaining autocrine TGF-β signaling controlling mycobacterial replication in infected-cells. We showed that while exogenous addition of increased concentration of TGF-β to infected-macrophages counteracted M. avium replication, pharmacological blockage of TGF-β receptor kinase activity with SB-431542 augmented bacterial load in infected-ECs. Moreover, the levels of TGF-β produced by ECs correlated with high and sustained levels of ERK1/2 activity. Inhibition of ERK1/2 activity with U0126 increased M. avium replication in infected-cells, suggesting that modulation of intracellular bacterial growth is dependent on the activation of ERK1/2. Interestingly, blockage of TGF-β receptor kinase activity with SB-431542 in infected-ECs inhibited ERK1/2 activity, enhanced intracellular M. avium burden and these effects were followed by a severe decrease in TGF-β production. In summary, our findings indicate that the amplitude of TGF-β signaling coordinates the strength and duration of ERK1/2 activity that is determinant for the control of intracellular mycobacterial growth.

  2. Competition for antigen between Th1 and Th2 responses determines the timing of the immune response switch during Mycobaterium avium subspecies paratuberulosis infection in ruminants.

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    Gesham Magombedze

    2014-01-01

    Full Text Available Johne's disease (JD, a persistent and slow progressing infection of ruminants such as cows and sheep, is caused by slow replicating bacilli Mycobacterium avium subspecies paratuberculosis (MAP infecting macrophages in the gut. Infected animals initially mount a cell-mediated CD4 T cell response against MAP which is characterized by the production of interferon gamma (Th1 response. Over time, Th1 response diminishes in most animals and antibody response to MAP antigens becomes dominant (Th2 response. The switch from Th1 to Th2 response occurs concomitantly with disease progression and shedding of the bacteria in feces. Mechanisms controlling this Th1/Th2 switch remain poorly understood. Because Th1 and Th2 responses are known to cross-inhibit each other, it is unclear why initially strong Th1 response is lost over time. Using a novel mathematical model of the immune response to MAP infection we show that the ability of extracellular bacteria to persist outside of macrophages naturally leads to switch of the cellular response to antibody production. Several additional mechanisms may also contribute to the timing of the Th1/Th2 switch including the rate of proliferation of Th1/Th2 responses at the site of infection, efficiency at which immune responses cross-inhibit each other, and the rate at which Th1 response becomes exhausted over time. Our basic model reasonably well explains four different kinetic patterns of the Th1/Th2 responses in MAP-infected sheep by variability in the initial bacterial dose and the efficiency of the MAP-specific T cell responses. Taken together, our novel mathematical model identifies factors of bacterial and host origin that drive kinetics of the immune response to MAP and provides the basis for testing the impact of vaccination or early treatment on the duration of infection.

  3. [Basic and clinical studies on pathogenesis of pulmonary Mycobacterium avium complex disease].

    Science.gov (United States)

    Suzuki, K

    1997-10-01

    I have studied pathogenesis of pulmonary Mycobacterium avium complex disease (PMAC), using mouse and human alveolar macrophage (PAM) model of the infection as well as clinical evaluations. The mouse model revealed no relation between natural resistance against the bacteria and the activation of macrophages which was evaluated on the basis of releasing capacities of prostaglandin E2 and superoxide anion. The PAM model suggested that TNF-alpha and GM-CSF could activate PAM to restrict the intracellular growth of the bacteria, probably not through the superoxide anion release, but through the myeloperoxidasae-halide system. It was also found that rifamycins in combination with clarithromycin could have a good bactericidal effect in the PAM-model of the infection. Clinical evaluations suggested that defect in local pulmonary defense, such as healed pulmonary tuberculous lesions, pneumoconiosis, and COPD was more important predisposing factor than defect in systemic defense in the development of PMAC. Most patients having PMAC without predisposing factors are elderly women, the reason of which is the most important question to be answered in the future studies.

  4. Phenolic compounds in cherry ( Prunus avium ) heartwood with a view to their use in cooperage.

    Science.gov (United States)

    Sanz, Miriam; Cadahía, Estrella; Esteruelas, Enrique; Muñoz, Angel Ma; Fernández De Simón, Brígida; Hernández, Teresa; Estrella, Isabel

    2010-04-28

    The phenolic and tannic composition of heartwood extracts from Prunus avium , commonly known as cherry tree, before and after toasting in cooperage were studied using HPLC-DAD and HPLC-DAD/ESI-MS. Nonflavonoid (16 compounds) and flavonoid (27 compounds) polyphenols were identified, 12 of them in only a tentative way. The nonflavonoids found were lignin constituents, and their pattern is different compared to oak, since they include compounds such as protocatechuic acid and aldehyde, p-coumaric acid, methyl vanillate, methyl syringate, and benzoic acid, but not ellagic acid, and only a small quantity of gallic acid. In seasoned wood we found a great variety of flavonoid compounds which have not been found in oak wood for cooperage, mainly, in addition to the flavan-3-ols (+)-catechin, a B-type procyanidin dimer, and a B-type procyanidin trimer, the flavanones naringenin, isosakuranetin, and eriodictyol and the flavanonols aromadendrin and taxifolin. Seasoned and toasted cherry wood showed different ratios of flavonoid to nonflavonoid compounds, since toasting results in the degradation of flavonoids, and the formation of nonflavonoids from lignin degradation. On the other hand, the absence of hydrolyzable tannins in cherry wood, which are very important in oak wood, is another particular characteristic of this wood that should be taken into account when considering its use in cooperage.

  5. Evaluation of physicochemical characteristics and antioxidant property of Prunus avium gum exudates.

    Science.gov (United States)

    Shabani, Hossein; Askari, Gholamreza; Jahanbin, Kambiz; Khodaeian, Faramarz

    2016-12-01

    In this study some physicochemical properties and elemental analysis of Prunus avium gum exudates were investigated. The gum studied had, on average, 75.14% carbohydrate, 11.3% uronic acids, 1.11% protein, 7.53% moisture content (w.b.) and 3.12% ash. Measured values for the angle of repose, Carr's index and Hausner ratio showed the good flow ability for the gum powder. The viscosity of 1% aqueous solution of the gum exhibited a Newtonian type of flow and with pH reduction the swelling index was increased. The average molecular weight of the main polysaccharide fraction was about 1.46×10(5)Da (146kDa). GC analysis showed that the main polysaccharide was composed of four kinds of neutral monosaccharides, namely mannose (Man), arabinose (Ara), galactose (Gal) and xylose (Xyl) with a relative molar ratio of 1.0:14.7:7.1:2.4. FTIR analysis showed the presence of carboxyl and hydroxyl groups and glycosidic linkage. The antioxidant activity of the gum was evaluated by determining DPPH scavenging and total phenolic contents which showed poor antioxidant property.

  6. [Identifying the S genotypes of sweet cherry (Prunus avium L.) cultivars].

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    Chen, Xiao-Liu; Chen, Xue-Sen; Shu, Huai-Rui

    2004-10-01

    This report identified S-RNase genes (S genes) of sweet cherry (Prunus avium L.), presented the sequences of S genes by using a pair of specific primers PruC2 and PruC4R based on the conserved regions C2 and RC4 of Rosaceous S-RNase genes and researched the S gene specific products from the genomic DNAs of different cultivars in which most of the S genotypese were unknown. The bands of PCR were cloned and their sequences were compared in GenBank. Four S genes were defined and the conclusion was made that all the same bands from PCR in the agarose gel had the same length and sequence of nucleic acid and were the same kind of S gene. The lengths of the amplified S genes are as follows: S1 is 677 bp, S3 762 bp, S4 945 bp, S6 456 bp. The S genotypes (S gene genotypes) of the tested self-incompatible cultivars were identified as follows: 'Hongdeng', 'Hongyan' and 'Early ruby', as same as 'Van', were S1 S3; 'Jueze', 'Hongfeng' and 'Napoleon' were S3S4; 'Dazi' was S1 S6; 'Changbahong' was S1 S4; 'Elton' was S3S6. The self-compatibile cultivars 'Waiyin No.7' and 'Stella' had the same S genotypes S3 S4'.

  7. High concentrations of anthocyanins in genuine cherry-juice of old local Austrian Prunus avium varieties.

    Science.gov (United States)

    Schüller, Elisabeth; Halbwirth, Heidi; Mikulic-Petkovsek, Maja; Slatnar, Ana; Veberic, Robert; Forneck, Astrid; Stich, Karl; Spornberger, Andreas

    2015-04-15

    Antioxidant activity and polyphenols were quantified in vapour-extracted juice of nine Austrian, partially endemic varieties of sweet cherry (Prunus avium): cv. 'Spätbraune von Purbach', cv. 'Early Rivers', cv. 'Joiser Einsiedekirsche', cv. 'Große Schwarze Knorpelkirsche' and four unidentified local varieties. Additionally the effect of storage was evaluated for six of the varieties. A variety showing the highest antioxidant capacity (9.64 μmol Trolox equivalents per mL), total polyphenols (2747 mg/L) and total cyanidins (1085 mg/L) was suitable for mechanical harvest and its juice did not show any losses of antioxidant capacity and total anthocyanin concentration during storage. The juice of cv. 'Große Schwarze Knorpelkirsche' had also high concentrations of total anthocyanins (873 mg/L), but showed substantial losses through storage. The local Austrian sweet cherry varieties from the Pannonian climate zone are particularly suitable for the production of processed products like cherry juice with high content of anthocyanins and polyphenols.

  8. In vitro plant regeneration from leaves and internode sections of sweet cherry cultivars (Prunus avium L.).

    Science.gov (United States)

    Matt, Andrea; Jehle, Johannes A

    2005-10-01

    Regeneration of adventitious shoots from leaves and, for the first time, from internode sections were compared and optimized for five economically important sweet cherry cultivars, i.e. "Schneiders", "Sweetheart", "Starking Hardy Giant", "Kordia" and "Regina" (Prunus avium L.). The influence of basal media, carbon source, combination and dosage of phytohormones, ethylene inhibitor such as silver thiosulfate and a 16 h:8 h light:dark photoperiod versus complete darkness were evaluated. Both, DKW/WPM (1:1) and Quoirin/Lepoivre (QL) basal media stimulated organogenesis more than QL/WPM (1:1), Chee and Pool (CP), Murashige Skoog (MS), Driver and Kuniyuki (DKW) or woody plant (WPM) media did. An induction phase in darkness resulted in lower or zero regeneration rates. The best regeneration efficiencies were generally obtained with thidiazuron in combination with indole-3-butyric-acid. The addition of silver thiosulfate resulted in a similar or reduced regeneration efficiency. Significant genotypic variability in adventitious bud formation was evident for both explant sources, leaf and internode section. Adventitious shoots were obtained from 11% of leaf explants and 50% of internode sections indicating that shoot regeneration from internodes was significantly more efficient than from leaves.

  9. Establishing Caenorhabditis elegans as a model for Mycobacterium avium subspecies hominissuis infection and intestinal colonization.

    Science.gov (United States)

    Everman, Jamie L; Ziaie, Navid R; Bechler, Jessica; Bermudez, Luiz E

    2015-09-24

    The nematode Caenorhabditis elegans has become a model system for studying the disease interaction between pathogens and the host. To determine whether the transparent nematode could serve as a useful model for Mycobacterium avium subspecies hominissuis (MAH) infection of the intestinal tract, worms were fed MAH and assayed for the effects of the bacterial infection on the worm. It was observed during feeding that viable MAH increases in the intestinal lumen in a time dependent manner. Ingestion of MAH was deemed non-toxic to worms as MAH-fed populations have similar survival curves to those fed E. coli strain OP50. Pulse-chase analysis using E. coli strain OP50 revealed that MAH colonize the intestinal tract, as viable MAH remain within the intestine after the assay. Visualization of intestinal MAH using histology and transmission electron microscopy demonstrates that MAH localizes to the intestinal lumen, as well as establishes direct contact with intestinal epithelium. Bacterial colonization appears to have a detrimental effect on the microvilli of the intestinal epithelial cells. The MAH ΔGPL/4B2 strain with a mutation in glycopeptidolipid production is deficient in binding to human epithelial cells (HEp-2), as well as deficient in its ability to bind to and colonize the intestinal tract of C. elegans as efficiently as wild-type MAH. These data indicate the C. elegans may serve as a useful model system for MAH pathogenesis and in determining the mechanisms used by MAH during infection and colonization of the intestinal epithelium.

  10. Short communication: effect of homogenization on heat inactivation of Mycobacterium avium subspecies paratuberculosis in milk.

    Science.gov (United States)

    Hammer, P; Kiesner, C; Walte, H-G C

    2014-01-01

    Mycobacterium avium ssp. paratuberculosis (MAP) can be present in cow milk and low numbers may survive high-temperature, short-time (HTST) pasteurization. Although HTST treatment leads to inactivation of at least 5 log10 cycles, it might become necessary to enhance the efficacy of HTST by additional treatments such as homogenization if the debate about the role of MAP in Crohn's disease of humans concludes that MAP is a zoonotic agent. This study aimed to determine whether disrupting the clumps of MAP in milk by homogenization during the heat treatment process would enhance the inactivation of MAP. We used HTST pasteurization in a continuous-flow pilot-plant pasteurizer and evaluated the effect of upstream, downstream, and in-hold homogenization on inactivation of MAP. Reduction of MAP at 72°C with a holding time of 28s was between 3.7 and 6.9 log10 cycles, with an overall mean of 5.5 log10 cycles. None of the 3 homogenization modes applied showed a statistically significant additional effect on the inactivation of MAP during HTST treatment.

  11. Search for Mycobacterium avium Subspecies paratuberculosis Antigens for the Diagnosis of Paratuberculosis

    Directory of Open Access Journals (Sweden)

    María Laura Mon

    2012-01-01

    Full Text Available The aim of this study was to evaluate a wide panel of antigens of Mycobacterium avium subsp. paratuberculosis (MAP to select candidates for the diagnosis of paratuberculosis (PTB. A total of 54 recombinant proteins were spotted onto nitrocellulose membranes and exposed to sera from animals with PTB (n=25, healthy animals (n=10, and animals experimentally infected with M. bovis (n=8. This initial screening allowed us to select seven antigens: MAP 2513, MAP 1693, MAP 2020, MAP 0038, MAP 1272, MAP 0209c, and MAP 0210c, which reacted with sera from animals with PTB and showed little cross-reactivity with sera from healthy animals and animals experimentally infected with M. bovis. The second step was to evaluate the antigen cocktail of these seven antigens by ELISA. For this evaluation, we used sera from animals with PTB (n=25, healthy animals (n=26, and animals experimentally infected with M. bovis (n=17. Using ELISA, the cocktail of the seven selected MAP antigens reacted with sera from 18 of the 25 animals with PTB and did not exhibit cross-reactivity with healthy animals and only low reactivity with animals with bovine tuberculosis. The combined application of these antigens could form part of a test which may help in the diagnosis of PTB.

  12. Mycobacterium avium ss. paratuberculosis Zoonosis - The Hundred Year War - Beyond Crohn's Disease.

    Science.gov (United States)

    Sechi, Leonardo A; Dow, Coad Thomas

    2015-01-01

    The factitive role of Mycobacterium avium ss. paratuberculosis (MAP) in Crohn's disease has been debated for more than a century. The controversy is due to the fact that Crohn's disease is so similar to a disease of MAP-infected ruminant animals, Johne's disease; and, though MAP can be readily detected in the infected ruminants, it is much more difficult to detect in humans. Molecular techniques that can detect MAP in pathologic Crohn's specimens as well as dedicated specialty labs successful in culturing MAP from Crohn's patients have provided strong argument for MAP's role in Crohn's disease. Perhaps more incriminating for MAP as a zoonotic agent is the increasing number of diseases with which MAP has been related: Blau syndrome, type 1 diabetes, Hashimoto thyroiditis, and multiple sclerosis. In this article, we debate about genetic susceptibility to mycobacterial infection and human exposure to MAP; moreover, it suggests that molecular mimicry between protein epitopes of MAP and human proteins is a likely bridge between infection and these autoimmune disorders.

  13. Mycobacterium avium ss. paratuberculosis Zoonosis – The Hundred Year War – Beyond Crohn’s Disease

    Science.gov (United States)

    Sechi, Leonardo A.; Dow, Coad Thomas

    2015-01-01

    The factitive role of Mycobacterium avium ss. paratuberculosis (MAP) in Crohn’s disease has been debated for more than a century. The controversy is due to the fact that Crohn’s disease is so similar to a disease of MAP-infected ruminant animals, Johne’s disease; and, though MAP can be readily detected in the infected ruminants, it is much more difficult to detect in humans. Molecular techniques that can detect MAP in pathologic Crohn’s specimens as well as dedicated specialty labs successful in culturing MAP from Crohn’s patients have provided strong argument for MAP’s role in Crohn’s disease. Perhaps more incriminating for MAP as a zoonotic agent is the increasing number of diseases with which MAP has been related: Blau syndrome, type 1 diabetes, Hashimoto thyroiditis, and multiple sclerosis. In this article, we debate about genetic susceptibility to mycobacterial infection and human exposure to MAP; moreover, it suggests that molecular mimicry between protein epitopes of MAP and human proteins is a likely bridge between infection and these autoimmune disorders. PMID:25788897

  14. Short communication: Viable Mycobacterium avium subspecies paratuberculosis in retail artisanal Coalho cheese from Northeastern Brazil.

    Science.gov (United States)

    Faria, A C S; Schwarz, D G G; Carvalho, I A; Rocha, B B; De Carvalho Castro, K N; Silva, M R; Moreira, M A S

    2014-07-01

    Mycobacterium avium ssp. paratuberculosis (MAP) is the etiologic agent of paratuberculosis and it potentially plays a role in Crohn's disease. In humans, the main route of transmission of MAP might be the intake of contaminated milk and dairy products. Considering that MAP has already been detected in many types of cheese in different counties, and that Coalho cheese is an important dairy product in northeastern Brazil, the aim of this study was to report the first detection of MAP in retail Coalho cheese in Brazil by PCR and culture. Of 30 retail Coalho cheese samples, 3 (10%) amplified fragments of a similar size to that expected (626 bp) were obtained and viable MAP was recovered by culture from 1 (3.3%) sample. The DNA from the positive culture sample was sequenced and showed 99% identity with the insertion sequence IS900 deposited in GenBank. It was possible to identify the presence of MAP-specific DNA in the analyzed samples for the first time in Brazil, and to recover viable cells from retail Coalho cheese.

  15. Pleural Mycobacterium Avium Complex Infection in an Immunocompetent Female with No Risk Factors

    Directory of Open Access Journals (Sweden)

    Ravi P. Manglani

    2015-01-01

    Full Text Available Mycobacterium avium complex (MAC infections rarely affect the pleura, accounting for 5–15% of pulmonary MAC. We report a case of MAC pleural effusion in an otherwise immunocompetent young patient. A 37-year-old healthy female with no past medical history was admitted to the hospital with two weeks of right sided pleuritic chest pain, productive cough, and fever. She was febrile, tachycardic, and tachypneic with signs of right sided pleural effusion which were confirmed by chest X-ray and chest CT. Thoracentesis revealed lymphocytic predominant exudative fluid. The patient underwent pleural biopsy, bronchoscopy with bronchoalveolar lavage, and video assisted thoracoscopic surgery (VATS, all of which failed to identify the causative organism. Six weeks later, MAC was identified in the pleural fluid and pleural biopsy by DNA hybridization and culture. The patient was started on clarithromycin, ethambutol, and rifampin. After six months of treatment, she was asymptomatic with complete radiological resolution of the effusion. The presence of lymphocytic effusion should raise the suspicion for both tuberculous and nontuberculous mycobacterial disease. Pleural biopsy must be considered to make the diagnosis. Clinicians must maintain a high index of suspicion of MAC infection in an otherwise immunocompetent patient presenting with a unilateral lymphocytic exudative effusion.

  16. Prevalence of Mycobacterium avium subsp. paratuberculosis in milk and dairy cattle in Southern Italy: preliminary results

    Directory of Open Access Journals (Sweden)

    Giacomo Marchetti

    2013-10-01

    Full Text Available Paratuberculosis affects all ruminants worldwide. Mycobacterium avium subsp. paratuberculosis (MAP could have a role in human diseases like Crohn’s. Some extra European Union (EU countries request importation of MAP-free products. Italy has not yet actualised a control programme and the diffusion of the infection is still unknown in Southern Italy. The aim of this study was to evaluate the prevalence of the infection in five regions of Southern Italy. Bulk tank milk and in-line milk filters were sampled in 780 dairy cattle herds and respectively analysed by ELISA and real time-polymerase chain reaction (PCR. One hundred and 55 out of 780 herds (19.9% were found positive by ELISA and/or real time PCR. Individual milk samples were then collected from all the producing animals of positive herds and from a selection of negative herds. The estimated prevalence varies from region to region between 2.8 and 5.5%. Our results indicate that the disease is widespread in the five regions. The observed prevalence could be underestimated.

  17. Is Primary Mycobacterium avium Complex Prophylaxis Necessary in Patients with CD4 cART?

    Science.gov (United States)

    Yangco, Bienvenido G.; Buchacz, Kate; Baker, Rose; Palella, Frank J.; Armon, Carl; Brooks, John T.

    2015-01-01

    We analyzed 369 patients with no prior Mycobacterium avium complex (MAC) infection and CD4 60 days and 19% had HIV RNA 10,000 copies/mL. Eleven patients had MAC infection within 6 months baseline (rate = 0.6/100 person months): 4/175 on MAC prophylaxis vs. 7/194, no MAC prophylaxis (p = 0.64). Of the 11 patients, seven had HIV RNA > 10,000, and three > 1000–9999 copies/mL at baseline (one missing). Median time to MAC infection was 62 days (IQR 43–126, maximum 139 days). No MAC infection occurred among 71 (19%) patients virologically suppressed (HIV RNA < 1000 copies/mL) at baseline, including 41 patients with no MAC prophylaxis during follow-up. A small number of eligible virologically suppressed participants and the lack of data on cART/MAC prophylaxis adherence limited our observational nonrandomized study. Primary MAC prophylaxis may not be required for cART-virologically suppressed patients with CD4 < 50 cells/mL. PMID:24833016

  18. Inferring biomarkers for Mycobacterium avium subsp. paratuberculosis infection and disease progression in cattle using experimental data

    Science.gov (United States)

    Magombedze, Gesham; Shiri, Tinevimbo; Eda, Shigetoshi; Stabel, Judy R.

    2017-03-01

    Available diagnostic assays for Mycobacterium avium subsp. paratuberculosis (MAP) have poor sensitivities and cannot detect early stages of infection, therefore, there is need to find new diagnostic markers for early infection detection and disease stages. We analyzed longitudinal IFN-γ, ELISA-antibody and fecal shedding experimental sensitivity scores for MAP infection detection and disease progression. We used both statistical methods and dynamic mathematical models to (i) evaluate the empirical assays (ii) infer and explain biological mechanisms that affect the time evolution of the biomarkers, and (iii) predict disease stages of 57 animals that were naturally infected with MAP. This analysis confirms that the fecal test is the best marker for disease progression and illustrates that Th1/Th2 (IFN-γ/ELISA antibodies) assays are important for infection detection, but cannot reliably predict persistent infections. Our results show that the theoretical simulated macrophage-based assay is a potential good diagnostic marker for MAP persistent infections and predictor of disease specific stages. We therefore recommend specifically designed experiments to test the use of a based assay in the diagnosis of MAP infections.

  19. Serodiagnosis of Mycobacterium avium complex disease in humans: translational research from basic mycobacteriology to clinical medicine.

    Science.gov (United States)

    Kobayashi, Kazuo

    2014-01-01

    Rapid and accurate diagnosis of infectious diseases, including mycobacterial disease such as tuberculosis (TB) and diseases due to nontuberculous mycobacteria (NTM), is a very important element of global health. The gold standard in diagnosis of mycobacterial diseases remains clinical examination, combined with direct microscopic examination of sputum and culture of bacteria. Culture of slowly growing mycobacteria, including Mycobacterium tuberculosis and NTM (such as M. avium complex: MAC), can take up to 4 to 6 weeks, and in 10-20% of cases the bacillus is not successfully cultivated. Diagnosis of MAC pulmonary disease (MAC-PD) is complicated and time-consuming (usually at least 1 month). I have characterized the nature of MAC antigens and immune responses from the aspect of basic mycobacteriology, and then translated to clinical science. My multicenter study in Japan has demonstrated the usefulness of a serodiagnostic test to determine serum IgA antibodies against mycobacterial glycopeptidolipid (GPL) core antigen for diagnosing MAC-PD within a few hours. To validate in a larger number of patients, at diverse geographic locations, and among other races, the test was also assessed the usefulness internationally in the United States and Taiwan. In this review, I discuss development of serodiagnosis of MAC-PD by translational research and international collaboration study.

  20. FurA contributes to the oxidative stress response regulation of Mycobacterium avium ssp. paratuberculosis.

    Science.gov (United States)

    Eckelt, Elke; Meißner, Thorsten; Meens, Jochen; Laarmann, Kristin; Nerlich, Andreas; Jarek, Michael; Weiss, Siegfried; Gerlach, Gerald-F; Goethe, Ralph

    2015-01-01

    The ferric uptake regulator A (FurA) is known to be involved in iron homeostasis and stress response in many bacteria. In mycobacteria the precise role of FurA is still unclear. In the presented study, we addressed the functional role of FurA in the ruminant pathogen Mycobacterium avium ssp. paratuberculosis (MAP) by construction of a furA deletion strain (MAPΔfurA). RNA deep sequencing revealed that the FurA regulon consists of repressed and activated genes associated to stress response or intracellular survival. Not a single gene related to metal homeostasis was affected by furA deletion. A decisive role of FurA during intracellular survival in macrophages was shown by significantly enhanced survival of MAPΔfurA compared to the wildtype, indicating that a principal task of mycobacterial FurA is oxidative stress response regulation in macrophages. This resistance was not associated with altered survival of mice after long term infection with MAP. Our results demonstrate for the first time, that mycobacterial FurA is not involved in the regulation of iron homeostasis. However, they provide strong evidence that FurA contributes to intracellular survival as an oxidative stress sensing regulator.

  1. Internalization-dependent recognition of Mycobacterium avium ssp. paratuberculosis by intestinal epithelial cells.

    Science.gov (United States)

    Pott, Johanna; Basler, Tina; Duerr, Claudia U; Rohde, Manfred; Goethe, Ralph; Hornef, Mathias W

    2009-12-01

    Mycobacterium avium ssp. paratuberculosis (MAP) is the causative agent of Johne's disease, a highly prevalent chronic intestinal infection in domestic and wildlife ruminants. The microbial pathogenesis of MAP infection has attracted additional attention due to an association with the human enteric inflammatory Crohn's disease. MAP is acquired by the faecal-oral route prompting us to study the interaction with differentiated intestinal epithelial cells. MAP was rapidly internalized and accumulated in a late endosomal compartment. In contrast to other opportunistic mycobacteria or M. bovis, MAP induced significant epithelial activation as indicated by a NF-kappaB-independent but Erk-dependent chemokine secretion. Surprisingly, MAP-induced chemokine production was completely internalization-dependent as inhibition of Rac-dependent bacterial uptake abolished epithelial activation. In accordance, innate immune recognition of MAP by differentiated intestinal epithelial cells occurred through the intracellularly localized pattern recognition receptors toll-like receptor 9 and NOD1 with signal transduction via the adaptor molecules MyD88 and RIP2. The internalization-dependent innate immune activation of intestinal epithelial cells is in contrast to the stimulation of professional phagocytes by extracellular bacterial constituents and might significantly contribute to the histopathological changes observed during enteric MAP infection.

  2. T-SPOT.TB Test(R) results in adults with Mycobacterium avium complex pulmonary disease.

    Science.gov (United States)

    Adams, Lisa V; Waddell, Richard D; Von Reyn, C Fordham

    2008-01-01

    The tuberculin skin test is limited by its inability to distinguish between infection with Mycobacterium tuberculosis and non-tuberculous mycobacteria (NTM). Newer interferon-gamma release assays using ESAT-6 and CFP-10 antigens should have a higher specificity for tuberculosis but have not been widely tested in adults with pulmonary disease due to NTM. In this study, we tested the T-SPOT.TB Test in patients with pulmonary disease due to Mycobacterium avium complex (MAC), the most common disease-causing NTM. Fourteen patients with prior culture-confirmed pulmonary disease due to MAC, 10 patients with prior culture-confirmed tuberculosis and 4 healthy controls were interviewed and tested with the T-SPOT.TB Test. 13 patients with MAC disease and 4 healthy subjects (negative controls) had non-reactive T-SPOT.TB results and 10 patients with prior tuberculosis (positive controls) had reactive results. One patient with MAC disease had a minimally reactive result on initial testing and a non-reactive result on re-testing. The T-SPOT.TB Test had a specificity of 94% for distinguishing between patients with prior MAC disease and prior tuberculosis disease, and will be useful in low tuberculosis prevalence settings where most mycobacterial infections are due to MAC. Reactions to the T-SPOT.TB Test may persist months to years after treatment of tuberculosis.

  3. The role of IL-10 in Mycobacterium avium subsp. paratuberculosis infection.

    Science.gov (United States)

    Hussain, Tariq; Shah, Syed Zahid Ali; Zhao, Deming; Sreevatsan, Srinand; Zhou, Xiangmei

    2016-12-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is an intracellular pathogen and is the causative agent of Johne's disease of domestic and wild ruminants. Johne's disease is characterized by chronic granulomatous enteritis leading to substantial economic losses to the livestock sector across the world. MAP persistently survives in phagocytic cells, most commonly in macrophages by disrupting its early antibacterial activity. MAP triggers several signaling pathways after attachment to pathogen recognition receptors (PRRs) of phagocytic cells. MAP adopts a survival strategy to escape the host defence mechanisms via the activation of mitogen-activated protein kinase (MAPK) pathway. The signaling mechanism initiated through toll like receptor 2 (TLR2) activates MAPK-p38 results in up-regulation of interleukin-10 (IL-10), and subsequent repression of inflammatory cytokines. The anti-inflammatory response of IL-10 is mediated through membrane-bound IL-10 receptors, leading to trans-phosphorylation and activation of Janus Kinase (JAK) family receptor-associated tyrosine kinases (TyKs), that promotes the activation of latent transcription factors, signal transducer and activators of transcription 3 (STAT3). IL-10 is an important inhibitory cytokine playing its role in blocking phagosome maturation and apoptosis. In the current review, we describe the importance of IL-10 in early phases of the MAP infection and regulatory mechanisms of the IL-10 dependent pathways in paratuberculosis. We also highlight the strategies to target IL-10, MAPK and STAT3 in other infections caused by intracellular pathogens.

  4. Attenuated strains of Mycobacterium avium subspecies paratuberculosis as vaccine candidates against Johne's disease.

    Science.gov (United States)

    Settles, Erik W; Kink, John A; Talaat, Adel

    2014-04-11

    Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is the causative agent of Johne's disease in ruminants. Johne's disease has a severe economic impact on the dairy industry in the USA and worldwide. In an effort to combat this disease, we screened several transposon mutants that were attenuated in the murine model of paratuberculosis for the potential use as live attenuated vaccines. Using the murine model, two vaccine candidates (pgs1360, pgs3965 with mutations of fabG2_2 and umaA1, respectively) were at or below the limit of detection for tissue colonization suggesting their low level persistence and hence safety. Prior to challenge, both candidates induced a M. paratuberculosis-specific IFN-γ, an indication of eliciting cell-mediated immunity. Following challenge with a virulent strain of M. paratuberculosis, the two vaccine candidates significantly reduced bacterial colonization in organs with reduced histological scores compared to control animals. In addition, one of the vaccine candidates (pgs3965) also induced IL-17a, a cytokine associated with protective immunity in mycobacterial infection. Our analysis suggested that the pgs3965 vaccine candidate is a potential live-attenuated vaccine that could be tested further in ruminant models of paratuberculosis. The analysis also validated our screening strategy to identify effective vaccine candidates against intracellular pathogens.

  5. Mycobacterium avium and purified protein derivative-specific cytotoxicity mediated by CD4+ lymphocytes from healthy HIV-seropositive and-seronegative individuals

    DEFF Research Database (Denmark)

    Ravn, P; Pedersen, B K

    1996-01-01

    mycobacteria. Our objective was to investigate the M.tuberculosis-and M. avium-specific cytotoxic capacity of T cells from healthy, bacille Calmette-Guérin-vaccinated, HIV-seropositive individuals. Blood mononuclear cells were obtained from 10 healthy HIV-seropositive and 10 healthy seronegative persons...... with no history of previous or active mycobacterial infection. Antigen-specific killing of macrophages presenting mycobacterial antigens (purified protein derivative or M. avium culture filtrate) was conducted. The phenotype of the killer cells was determined by a fluorescence-activated cell sorter after antigen...

  6. The investigation of the truncated mbtA gene within the mycobactin cluster of Mycobacterium avium subspecies paratuberculosis as a novel diagnostic marker for real-time PCR.

    Science.gov (United States)

    de Kruijf, Marcel; Coffey, Aidan; O'Mahony, Jim

    2017-05-01

    The inability of Mycobacterium avium subspecies paratuberculosis (MAP) to produce endogenous mycobactin in-vitro is most likely due to the presence of a truncated mbtA gene within the mycobactin cluster of MAP. The main goal of this study was to investigate this unique mbtA truncation as a potential novel PCR diagnostic marker for MAP. Novel primers were designed that were located within the truncated region and the contiguous MAP2179 gene. Primers were evaluated against non-MAP isolates and no amplicons were generated. The detection limit of this mbtA-MAP2179 target was evaluated using a range of MAP DNA concentrations, MAP inoculated faecal material and 20 MAP isolates. The performance of mbtA-MAP2179 was compared to the established f57 target. The detection limits recorded for MAP K-10 DNA and from MAP K-10 inoculated faecal samples were 0.34pg and 10(4)CFU/g respectively for both f57 and mbtA-MAP2179. A detection limit of 10(3)CFU/g was recorded for both targets, but not achieved consistently. The detection limit of MAP from inoculated faecal material was successful at 10(3)CFU/g for mbtA-MAP2179 when FAM probe real-time PCR was used. A MAP cell concentration of 10(2)CFU/g was detected successfully, but again not consistently achieved. All 20 mycobacterial isolates were successfully identified as MAP by f57 and mbtA-MAP2179. Interestingly, the mbtA-MAP2179 real-time PCR assay resulted in the formation of a unique melting curve profile that contained two melting curve peaks rather than one single peak. This melting curve phenomenon was attributed towards the asymmetrical GC% distribution within the mbtA-MAP2179 amplicon. This study investigated the implementation of the mbtA-MAP2179 target as a novel diagnostic marker and the detection limits obtained with mbtA-MAP2179 were comparable to the established f57 target, making the mbtA-MAP2179 an adequate confirmatory target. Moreover, the mbtA-MAP2179 target could be implemented in multiplex real-time PCR assays and

  7. Pengembangan Enzyme-Linked Immunosorbent Assay Paratuberkulosis dengan Antigen Protoplasmik Mycobacterium avium Subspecies Paratuberculosis Isolat Lapang (DEVELOPMENT OF PARATUBERCULOSIS ENZYME-LINKED IMMUNO-SORBENT ASSAY WITH PROTOPLASMIC ANTIGEN OF MYCO

    Directory of Open Access Journals (Sweden)

    Rahmat Setya Adji

    2015-08-01

    Full Text Available Enzyme-linked immunosorbent assay (ELISA is a serological test method most widely used for thediagnosis of paratuberculosis, because it has a better sensitivity compared to other serological test.Protoplasmic antigen (PPA or cellular extract is still the main choice for the diagnosis of paratuberkulosisdevelopment. The aim of research was to use the PPA Mycobacterium avium subspeciesparatuberculosis(MAP field isolates for the development of paratuberculosis ELISA (ELISA PPA-L. As many as 322cattle sera (300 negative and 22 positive were tested using this method and compared with IDEXXcommercial kit. The sensitivity and specificity of ELISA PPA-L test results were 68.18% and 97.0%,whereas for the IDEXX kit were 63.64% and 97.33%respectively. ELISA PPA-L had higher sensitivity andlower specificity compared to the IDEXX commercial kit. ELISA test using protoplasmic antigen of MAPfield isolates has good ability for paratuberculosis serological test and can be used for screening test of thedisease in Indonesia.

  8. Original Thinking

    Directory of Open Access Journals (Sweden)

    Ashok Natarajan

    2012-04-01

    Full Text Available History that comes to us as a chronology of events is really a collective existence that is evolving through several stages to develop Individuality in all members of the society. The human community, nation states, linguistic groups, local castes and classes, and families are the intermediate stages in development of the Individual. The social process moves through phases of survival, growth, development and evolution. In the process it organizes the consciousness of its members at successive levels from social external manners, formed behavior, value-based character and personality to culminate in the development of Individuality. Through this process, society evolves from physicality to Mentality. The power of accomplishment in society and its members develops progressively through stages of skill, capacity, talent, and ability. Original thinking is made possible by the prior development of thinking that organizes facts into information. The immediate result of the last world war was a shift in reliance from physical force and action to mental conception and mental activity on a global scale. At such times no problem need defy solution, if only humanity recognizes the occasion for thinking and Original Thinking. The apparently insoluble problems we confront are an opportunity to formulate a comprehensive theory of social evolution. The immediate possibility is to devise complete solutions to all existing problems, if only we use the right method of thought development.

  9. Anaerobic facultative bacteria isolated from the gut of rabbits fed different diets.

    Science.gov (United States)

    Canganella, F; Zirletta, G; Gualterio, L; Massa, S; Trovatelli, L D

    1992-11-01

    Anaerobic facultative bacteria colonizing the intestinal tract of conventional rabbits fed three different diets (standard pellet, hay and pellet/hay mixture) were enumerated in brain heart infusion agar. Colony counts recovered from homogenized samples of small intestine, caecum and rectum differed with reference to the diet given. Among anaerobic groups, identified from rabbit fed pellet/hay mixture, Enterococci (E. faecalis, E. avium, E. faecium and E. durans) represented the predominant flora. Enterobacters (E. cloacae and E. aerogenes) accounted for about 10 to 25% of the bacteria in the rectum and colon respectively, whereas Staphylococci (S. intermedius, S. epidermidis and S. lentus) represented 11% of the bacteria isolated from colon.

  10. Non-tuberculous mycobacteria isolated from slaughter pigs in Mubende district, Uganda

    Directory of Open Access Journals (Sweden)

    Muwonge Adrian

    2012-05-01

    Full Text Available Abstract Background The importance of infections caused by non-tuberculous mycobacteria (NTM in animals and humans has gained considerable recognition during the past few years. In the developed world, where pig production is extensively practiced, studies on mycobacterial infections and related control strategies have received increasing attention. The infections are reported to be caused by a wide spectrum of NTM. Unfortunately, these infections have been less recognized in sub-Saharan Africa owing to lack of awareness and systematic studies. In this study we aimed at isolating and identifying species of mycobacteria involved in causing infections in slaughter pigs in Mubende district of Uganda. Furthermore we wanted to identify factors associated with infection prevalence in the study area. Methods A total of 363 lymph nodes were collected and cultured for the presence of mycobacteria. Isolates were identified by 16S rDNA gene sequencing. A questionnaire survey was administered to identify production related factors associated with infection prevalence. Data were assembled and analysed using descriptive statistics and mixed effects logistic regression analysis. Results Mycobacteria were detected in 39 % (143/363 of the examined lymph nodes, 63 % (59/93 of lymph nodes with gross lesions typical of mycobacteriosis and 31% (84/270 of lymph nodes with no visible lesions. Nineteen per cent of the isolated mycobacteria were identified as Mycobacterium (M avium, of these 78% and 22% were M. avium sub sp. Hominissuis and avium respectively. Other mycobacterial species included M. senuense (16%, M. terrae (7% and M. asiaticum (6%. This study found free range systems (OR = 3.0; P = 0.034 and use of water from valley dams (OR = 2.0; P = 0.049 as factors associated with high prevalence of mycobacteria in slaughter pigs. Conclusions This study demonstrated a high prevalence of NTM infections among slaughter pigs in Mubende district of

  11. Serologic tests for detecting antibodies against Mycobacterium bovis and Mycobacterium avium subspecies paratuberculosis in Eurasian wild boar (Sus scrofa scrofa).

    Science.gov (United States)

    Boadella, Mariana; Lyashchenko, Konstantin; Greenwald, Reena; Esfandiari, Javan; Jaroso, Raquel; Carta, Tania; Garrido, Joseba M; Vicente, Joaquín; de la Fuente, José; Gortázar, Christian

    2011-01-01

    New tools to detect exposure of free-range Eurasian wild boar (Sus scrofa scrofa) to pathogenic mycobacteria would be valuable for improved disease surveillance and wildlife management. Two hundred sera from wild boar of known Mycobacterium bovis infection status were used to evaluate test suitability for the detection of antibodies against M. bovis and Mycobacterium avium subsp. paratuberculosis (or cross-reacting members of the M. avium complex). Two traditional enzyme-linked immunosorbent assays were evaluated using M. bovis purified protein derivative (bPPD) and paratuberculosis protoplasmatic antigen 3 (PPA3) as antigens, respectively, and a new point-of-care test format for bovine tuberculosis (bTB) that uses the innovative dual-path platform (DPP TB) test. The effect of individual factors (sex, age, lesions) on the diagnostic performance of the serologic tests was also determined. Although the DPP had a sensitivity of 89.6% and a specificity of 90.4%, for bPPD, the sensitivity was 79.2% and the specificity 100%. Both tests had a kappa agreement of 0.80. Sixty-five of 68 (95.6%) wild boar sera with antibodies against the PPA3 antigen corresponded to known M. bovis-infected wild boar. Significant differences were not observed in the bPPD and DPP readings among lesion categories or between age classes. A slight sex-related difference in sensitivity toward males in the DPP was found, but it was not detected in the bPPD enzyme-linked immunosorbent assay. The results support the use of antibody-based diagnostic tests for both large-scale and individual bTB testing of Eurasian wild boar and suggest that wild boar cannot be used as sentinels for infections caused by M. avium complex members.

  12. Characterization of Pseudomonas pathovars isolated from rosaceous fruit trees in East Algeria.

    Science.gov (United States)

    Harzallah, D; Sadallah, S; Larous, L

    2004-01-01

    A survey of bacterial diseases due to Pseudomonas on rosaceous fruit trees was conducted. In forty two orchards located in the Constantine region ( East Algeria). Pseudomonas isolates were identified on the bases of their cultural and biochemical characteristics . A total of fifty nine phytopathogenic bacteria were isolated from diseased pome and stone fruit trees. Thirty one strains comparable to Pseudomonas syringae pv. syringae were isolated from cherry (Prunus avium L.), plum (P. domestica L.), apricot (P. armeniaca L.), almond (P. dulcis L.) and pear trees (Pirus communis L.); sixteen strains comparable to Pseudomonas syringae pv. morsprunorum were obtained from samples of cherry and plum. Twelve strains of Pseudomonas viridiflava were isolated from cherry, apricot and peach (Prunus persica L.).

  13. Novel antigens used to detect cell-mediated immune responses over time in Mycobacterium avium subsp. paratuberculosis infected cattle

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Aagaard, Claus; Nielsen, Søren Saxmose;

    Early stage Mycobacterium avium subsp. paratuberculosis (MAP) infection of cattle can be detected by measuring specific cell mediated immune responses, using the interferon gamma (IFN-γ) test. Available IFN-γ tests are using purified protein derivatives of MAP (PPDj) which are crude products...... on the same 30 heifers from a known MAP infected herd. Determination of cut-off for each antigen was based on samples from a non-infected herd, including 60 heifers. Based on PPDj stimulations, more than 50% of the heifers tested MAP positive at the first two samplings, whereas only 20% tested positive...

  14. Physicochemical characteristics, antioxidant activity, organic acid and sugar contents of 12 sweet cherry (Prunus avium L.) cultivars grown in Turkey.

    Science.gov (United States)

    Hayaloglu, Ali Adnan; Demir, Nurullah

    2015-03-01

    Physical characteristics, antioxidant activity and chemical constituents of 12 cultivars (Prunus avium L.) of sweet cherry (Belge, Bing, Dalbasti, Durona di Cesena, Lambert, Merton Late, Starks Gold, Summit, Sweetheart, Van, Vista, and 0-900 Ziraat) were investigated. Significant differences (P cherries. Four different sugars were observed in the samples and their concentrations ordered as glucose > fructose > sucrose > xylose. Sugar alcohol in the cherries was represented by sorbitol (more than 90%) and its concentration varied between 13.93 and 27.12 g/kg. As a result significant differences were observed among the physical properties and chemical constituents of the cherry cultivars.

  15. Early-screening for resistance to Phytophthora sp.p. in wild cherry clones (Prunus avium L.

    Directory of Open Access Journals (Sweden)

    Guerri S

    2004-01-01

    Full Text Available A new method for early selection of wild cherry clones for resistance to Phytophthora sp. is presented. Four Phytophthora species (P. cinnamomi, P. citrophthora, P. megasperma, P. alni were tested in vitro on four micropropagated cherry (Prunus avium clones, obtaining reliable and reproducible results. Variability in clones susceptibilities and in parasite virulence has been evidenced. Phytophthora citrophthora is confirmed to be a dangerous parasite, due to its capability to produce symptoms rapidly and its fitness to Mediterranean environments. On the contrary P. alni does not seem a dangerous parasite for wild cherry.

  16. Characterization of the long-term immune response to vaccination against Mycobacterium avium subsp. paratuberculosis in Danish dairy cows

    DEFF Research Database (Denmark)

    Thomsen, Vibeke Thulstrup; Nielsen, Søren Saxmose; Thakur, Aneesh;

    2012-01-01

    of vaccination on the cell-mediated immune response and to evaluate a possible interference with the diagnosis of M. bovis infections.The results showed that 37% of samples from vaccinated animals and 5% of samples from non-vaccinated animals, respectively, were test positive in the milk antibody ELISA......Vaccination of cattle against Mycobacterium avium subsp. paratuberculosis (MAP) provides partial protection by delayed shedding of MAP and reduced numbers of clinically affected animals. The duration of vaccine induced immune response is not known. The primary objective of this study was therefore...

  17. Isolation of nontuberculous mycobacteria (NTM) from household water and shower aerosols in patients with pulmonary disease caused by NTM.

    Science.gov (United States)

    Thomson, Rachel; Tolson, Carla; Carter, Robyn; Coulter, Chris; Huygens, Flavia; Hargreaves, Megan

    2013-09-01

    It has been postulated that susceptible individuals may acquire infection with nontuberculous mycobacteria (NTM) from water and aerosol exposure. This study examined household water and shower aerosols of patients with NTM pulmonary disease. The mycobacteria isolated from clinical samples from 20 patients included M. avium (5 patients), M. intracellulare (12 patients), M. abscessus (7 patients), M. gordonae (1 patient), M. lentiflavum (1 patient), M. fortuitum (1 patient), M. peregrinum (1 patient), M. chelonae (1 patient), M. triplex (1 patient), and M. kansasii (1 patient). One-liter water samples and swabs were collected from all taps, and swimming pools or rainwater tanks. Shower aerosols were sampled using Andersen six-stage cascade impactors. For a subgroup of patients, real-time PCR was performed and high-resolution melt profiles were compared to those of ATCC control strains. Pathogenic mycobacteria were isolated from 19 homes. Species identified in the home matched that found in the patient in seven (35%) cases: M. abscessus (3 cases), M. avium (1 case), M. gordonae (1 case), M. lentiflavum (1 case), and M. kansasii (1 case). In an additional patient with M. abscessus infection, this species was isolated from potable water supplying her home. NTM grown from aerosols included M. abscessus (3 homes), M. gordonae (2 homes), M. kansasii (1 home), M. fortuitum complex (4 homes), M. mucogenicum (1 home), and M. wolinskyi (1 home). NTM causing human disease can be isolated from household water and aerosols. The evidence appears strongest for M. avium, M. kansasii, M. lentiflavum, and M. abscessus. Despite a predominance of disease due to M. intracellulare, we found no evidence for acquisition of infection from household water for this species.

  18. Distribution and fine-scale spatial-genetic structure in British wild cherry (Prunus avium L.).

    Science.gov (United States)

    Vaughan, S P; Cottrell, J E; Moodley, D J; Connolly, T; Russell, K

    2007-05-01

    Insights into the within-population spatial-genetic structure (SGS) of forest tree species, where little is known regarding seed and pollen dispersal patterns, enhance understanding of their ecology and provide information of value in conservation and breeding. This study utilised 13 polymorphic simple sequence repeat loci to investigate the impact of asexual recruitment, management regime and tree size on the development of SGS in wild cherry (Prunus avium L). Only 246 genotypes were identified in the 551 trees sampled, reflecting significant levels of clonal reproduction in both managed and unmanaged populations. Naturally regenerated wild cherry was spatially aggregated under both management regimes. However, in the managed population, sexually derived trees accounted for a greater proportion of the smaller size classes, whereas vegetatively produced trees dominated the smaller size classes in the unmanaged population. High overall SGS values (Sp 0.030-Sp 0.045) were observed when considering only sexually derived genets and kinship coefficients were significant up to the 120 m distance class for both populations. The inclusion of clonal ramets in the analysis significantly increased the overall SGS (Sp 0.089-Sp 0.119) as well as kinship coefficients in the 40-80 m distance classes, illustrating the dramatic impact of vegetative propagation on SGS in this species. Increased spatial aggregation and regeneration appeared to be concomitant with increased SGS in the 40 m distance class in the unmanaged population. Neighbourhood size estimates were relatively small for both populations and kinship coefficients were found to decline with distance under both management regimes, suggesting that common mechanisms may restrict gene dispersal in wild cherry.

  19. [Characteristics of urea 15N absorption, allocation, and utilization by sweet-cherry (Prunus avium L.)].

    Science.gov (United States)

    Zhao, Feng-Xia; Jiang, Yuan-Mao; Peng, Fu-Tian; Gao, Xiang-Bin; Liu, Bing-Hua; Wang, Hai-Yun; Zhao, Lin

    2008-03-01

    With five-year old 'Zaodaguo' sweet-cherry (Prunus avium L.) as test material, this paper studied the characteristics of its urea 15N absorption, allocation, and utilization when applied before bud-break. The results showed that the Ndff of different organs increased gradually with time, and was higher in fine roots and storage organs at full-blooming stage. At fruit core-hardening stage, the Ndff of long shoots and leaves increased quickly, reaching to 0.72% and 0.59% , respectively. From fruit core-hardening to harvesting stage, the Ndff of fruit had a rapid increase, with the peak (1.78%) at harvesting stage. After harvest, the Ndff of neonatal organs increased slowly while that of storage organs increased quickly. At full-blooming stage, the absorbed 15N in roots was firstly allocated to storage organs, with the highest allocation rate (54.91%) in large roots. At fruit core-hardening stage, the allocation rate in fine roots and storage organs decreased from 85.43% to 55.11%, while that in neonatal branches and leaves increased to 44.89%. At harvesting stage, the allocation rate in different organs had no significant change, but after harvest, the absorbed 15N had a rapid translocation to storage organs, and the allocation rate in fine roots and storage organs reached the highest (72.26%) at flower bud differentiation stage. The 15N allocation rate in neonatal branches and leaves at flower bud differentiation stage was decreased by 19.31%, compared with that at harvesting stage. From full-blooming to flower bud differentiation stage, the utilization rate of urea 15N was increasing, and reached the peak (16.86%) at flower bud differentiation stage.

  20. Dormancy in sweet cherry (Prunus avium L. under Mediterranean climatic conditions

    Directory of Open Access Journals (Sweden)

    Ahmed Mahhou

    2007-01-01

    Full Text Available It is admitted that the lack of winter chilling is a limiting factor for the cultivation of temperate fruit trees in warm climates. Nevertheless, the characteristics of dormancy in sweet cherry under such conditions are still not fully understood. Therefore, and in order to contribute to the elucidation of these mechanisms, the objective of this work is to evaluate the dormancy of sweet cherry (Prunus avium L. under the Mediterranean conditions of Meknes region (Morocco. Evaluation of dormancy behaviour of four varieties was made on the basis of a biological test, known as the « single node cuttings », and histological dissections of flower buds at the time of bud break. The variations of mean time to bud break (MTB were interpreted in terms of the evolution of growth capacity of the buds. The lack of cold affected tree phenological development. The evolution of dormancy in cherry buds showed three phases with variable levels of inertia: the first and third phases being of similar intensity and duration while the second phase was of a high level of inertia (endo-dormancy. The correlative inhibitions of the leaves imposed on the buds continued up to the arrival of continuous low temperatures. The delay in the decrease of inertia reveals insufficient chilling accumulation causing incomplete release from dormancy and low bud break percentage in the orchard. Histological observations showed that vascular connection of flower buds was established during bud break stage. However, flowers showed certain abnormalities, ie low pollen production and malformations of pistils, which limit the possibilities of pollination.

  1. Evaluation of evapotranspiration estimation methods for sweet cherry trees (Prunus avium) in sub-humid climate.

    Science.gov (United States)

    Denmirtas, Cigdem; Buyukcangaz, Hakan; Yazgan, Senih; Candogan, Burak Nazmi

    2007-02-01

    This study was carried out in the summer of 2001 in a 3 year old and in the summer of 2002 in a 4 year old sweet cherry trees (Prunus avium, variety Z-900) on Mazzard rootstocks in Bayramic-Canakkale which is located in the west part of Turkey. Micro-sprinkler irrigation was selected as the irrigation method. The trees were subjected to four micro-sprinkler irrigation treatments (T-1, T-2, T-3 and T-4). The water applied in treatment T-3 was considered sufficient to satisfy fully needs of the crop (100% of ETc) and to allow good rooting and tree growth. The water balance relationship was used in estimating ETc. A total of 4 climatological methods were selected for estimating reference crop evapotranspiration on a daily basis. Some of these methods are based on combination theory and others are empirical methods based primarily on solar radiation, temperature ans relative humidity. An attempt was made in the current study to develop regional relationship between the evapotranspiration measured and that estimated by the climatological methods, such as FAO-Penman, Penman-Monteith, FAO-Radiation and FAO-Blaney-Criddle. Performance of the climatological methods in estimating the ETo values as compared to the measured values was evaluated on the basis of root mean square error (RMSE). In 2001, the Penman-Monteith equation gave the best results followed by FAO-Penman, FAO-Radiation and FAO-Blaney-Criddle. In 2002, the Penman-Monteith and FAO-Blaney-Criddle equations gave same results.

  2. Heterozygote excess in a self-incompatible and partially clonal forest tree species -- Prunus avium L.

    Science.gov (United States)

    Stoeckel, Solenn; Grange, Jérôme; Fernández-Manjarres, Juan F; Bilger, Isabelle; Frascaria-Lacoste, Nathalie; Mariette, Stéphanie

    2006-07-01

    Wild cherry (Prunus avium L.), a partially asexual self-incompatible forest tree, shows heterozygote excess, which is a poorly studied phenomenon. In three natural populations, we found significant heterozygote excess at almost all investigated loci (eight microsatellites and markers for the self-incompatibility locus). We examined four hypotheses to account for this observed heterozygote excess. First, negative F(IS) can result from a lack of selfed progeny in small populations of outcrossing species. A second explanation for negative F(IS) is selection during the life cycle of the most heterozygous individuals. A third explanation is negative assortative mating when reproduction occurs between individuals bearing phenotypes more dissimilar than by chance. The last explanation for negative F(IS) relies on asexual reproduction. Expectations for each hypothesis were tested using empirical data. Patterns of F(IS) differed among loci. Nevertheless, our experimental results did not confirm the small sample size hypothesis. Although one locus is probably under a hitch-hiking effect from the SI locus, we rejected the effect of the self-incompatibility locus for the genome as a whole. Similarly, although one locus showed a clear pattern consistent with the selection of heterozygous individuals, the heterosis effect over the whole genome was rejected. Finally, our results revealed that clonality probably explains significant negative F(IS) in wild cherry populations when considering all individuals. More theoretical effort is needed to develop expectations and hypotheses, and test them in the case of species combining self-incompatibility and partially asexual reproduction.

  3. Management of genetic resources in the nursery system of wild cherry (Prunus avium L.

    Directory of Open Access Journals (Sweden)

    Proietti R

    2006-01-01

    Full Text Available Knowledge of genetic and adaptive traits of reproductive materials used in the nursery system of wild cherry, could be an useful instrument to improve ecological and economic sustainability of plantation ecosystems. This work reports results from a research which the objectives were: 1 to study the genetic variation of a Prunus avium L. Population, used for seed harvesting, through its multi-locus genotypes detected by starch gel electrophoresis; 2 to analyze the level of genetic variation within and among different steps in a commercial nursery system (basic population and sub-populations, seedlings aged S1T1 and S1T2, plantation. Results showed low genetic variation levels of the basic population, similar to a reference system of other 12 wild cherry Italian populations and to other French and Caucasian materials. The genetic distances among Monte Baldo and some closer Lombardy provenances (Area Garda, Bosco Fontana, Valtellina were smaller than the Venice Region populations (Monti Lessini and Asiago. Number of alleles and percentage of polymorphic loci within the complex of Monte Baldo provenance and multiplication materials were similar, whilst a variable value of Fis was noted. Indeed, along with the nursery system until the plantation, heterozygosis initially (S1T1 increased, then decreased proceeding to the plantation. This fluctuation of FIS values could be determined by seed lots characterized initially by higher levels of variation, due to self-incompatibility. In the following steps, a possible selection pressure can affect randomly the genotypic structure of wild cherry by increasing the homozygosity. There is not among population a well defined geographic characterization, as suggested by genetic distances, therefore homogeneous seed harvest could be established an area larger than geographic and administrative borders. On this way we could have reproductive material with a wide genetic base and environmental adaptability. To

  4. Evaluation of a PCR assay on overgrown environmental samples cultured for Mycobacterium avium subsp. paratuberculosis.

    Science.gov (United States)

    Arango-Sabogal, Juan C; Labrecque, Olivia; Paré, Julie; Fairbrother, Julie-Hélène; Roy, Jean-Philippe; Wellemans, Vincent; Fecteau, Gilles

    2016-11-01

    Culture of Mycobacterium avium subsp. paratuberculosis (MAP) is the definitive antemortem test method for paratuberculosis. Microbial overgrowth is a challenge for MAP culture, as it complicates, delays, and increases the cost of the process. Additionally, herd status determination is impeded when noninterpretable (NI) results are obtained. The performance of PCR is comparable to fecal culture, thus it may be a complementary detection tool to classify NI samples. Our study aimed to determine if MAP DNA can be identified by PCR performed on NI environmental samples and to evaluate the performance of PCR before and after the culture of these samples in liquid media. A total of 154 environmental samples (62 NI, 62 negative, and 30 positive) were analyzed by PCR before being incubated in an automated system. Growth was confirmed by acid-fast bacilli stain and then the same PCR method was again applied on incubated samples, regardless of culture and stain results. Change in MAP DNA after incubation was assessed by converting the PCR quantification cycle (Cq) values into fold change using the 2(-ΔCq) method (ΔCq = Cq after culture - Cq before culture). A total of 1.6% (standard error [SE] = 1.6) of the NI environmental samples had detectable MAP DNA. The PCR had a significantly better performance when applied after culture than before culture (p = 0.004). After culture, a 66-fold change (SE = 17.1) in MAP DNA was observed on average. Performing a PCR on NI samples improves MAP culturing. The PCR method used in our study is a reliable and consistent method to classify NI environmental samples.

  5. Establishing Caenorhabditis elegans as a model for Mycobacterium avium subspecies hominissuis infection and intestinal colonization

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    Jamie L. Everman

    2015-10-01

    Full Text Available The nematode Caenorhabditis elegans has become a model system for studying the disease interaction between pathogens and the host. To determine whether the transparent nematode could serve as a useful model for Mycobacterium avium subspecies hominissuis (MAH infection of the intestinal tract, worms were fed MAH and assayed for the effects of the bacterial infection on the worm. It was observed during feeding that viable MAH increases in the intestinal lumen in a time dependent manner. Ingestion of MAH was deemed non-toxic to worms as MAH-fed populations have similar survival curves to those fed E. coli strain OP50. Pulse-chase analysis using E. coli strain OP50 revealed that MAH colonize the intestinal tract, as viable MAH remain within the intestine after the assay. Visualization of intestinal MAH using histology and transmission electron microscopy demonstrates that MAH localizes to the intestinal lumen, as well as establishes direct contact with intestinal epithelium. Bacterial colonization appears to have a detrimental effect on the microvilli of the intestinal epithelial cells. The MAH ΔGPL/4B2 strain with a mutation in glycopeptidolipid production is deficient in binding to human epithelial cells (HEp-2, as well as deficient in its ability to bind to and colonize the intestinal tract of C. elegans as efficiently as wild-type MAH. These data indicate the C. elegans may serve as a useful model system for MAH pathogenesis and in determining the mechanisms used by MAH during infection and colonization of the intestinal epithelium.

  6. Transcriptional Profiling of Ileocecal Valve of Holstein Dairy Cows Infected with Mycobacterium avium subsp. Paratuberculosis.

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    Randy J Hempel

    Full Text Available Johne's disease is a chronic infection of the small intestine caused by Mycobacterium avium subspecies paratuberculosis (MAP, an intracellular bacterium. The events of pathogen survival within the host cell(s, chronic inflammation and the progression from asymptomatic subclinical stage to an advanced clinical stage of infection, are poorly understood. This study examines gene expression in the ileocecal valve (ICV of Holstein dairy cows at different stages of MAP infection. The ICV is known to be a primary site of MAP colonization and provides an ideal location to identify genes that are relevant to the progression of this disease. RNA was prepared from ICV tissues and RNA-Seq was used to compare gene transcription between clinical, subclinical, and uninfected control animals. Interpretation of the gene expression data was performed using pathway analysis and gene ontology categories containing multiple differentially expressed genes. Results demonstrated that many of the pathways that had strong differential gene expression between uninfected control and clinical cows were related to the immune system, such as the T- and B-cell receptor signaling, apoptosis, NOD-like receptor signaling, and leukocyte transendothelial migration pathways. In contrast, the comparison of gene transcription between control and subclinical cows identified pathways that were primarily involved in metabolism. The results from the comparison between clinical and subclinical animals indicate recruitment of neutrophils, up regulation of lysosomal peptidases, increase in immune cell transendothelial migration, and modifications of the extracelluar matrix. This study provides important insight into how cattle respond to a natural MAP infection at the gene transcription level within a key target tissue for infection.

  7. Transcriptional Profiling of Ileocecal Valve of Holstein Dairy Cows Infected with Mycobacterium avium subsp. Paratuberculosis.

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    Hempel, Randy J; Bannantine, John P; Stabel, Judith R

    2016-01-01

    Johne's disease is a chronic infection of the small intestine caused by Mycobacterium avium subspecies paratuberculosis (MAP), an intracellular bacterium. The events of pathogen survival within the host cell(s), chronic inflammation and the progression from asymptomatic subclinical stage to an advanced clinical stage of infection, are poorly understood. This study examines gene expression in the ileocecal valve (ICV) of Holstein dairy cows at different stages of MAP infection. The ICV is known to be a primary site of MAP colonization and provides an ideal location to identify genes that are relevant to the progression of this disease. RNA was prepared from ICV tissues and RNA-Seq was used to compare gene transcription between clinical, subclinical, and uninfected control animals. Interpretation of the gene expression data was performed using pathway analysis and gene ontology categories containing multiple differentially expressed genes. Results demonstrated that many of the pathways that had strong differential gene expression between uninfected control and clinical cows were related to the immune system, such as the T- and B-cell receptor signaling, apoptosis, NOD-like receptor signaling, and leukocyte transendothelial migration pathways. In contrast, the comparison of gene transcription between control and subclinical cows identified pathways that were primarily involved in metabolism. The results from the comparison between clinical and subclinical animals indicate recruitment of neutrophils, up regulation of lysosomal peptidases, increase in immune cell transendothelial migration, and modifications of the extracelluar matrix. This study provides important insight into how cattle respond to a natural MAP infection at the gene transcription level within a key target tissue for infection.

  8. Genetic loci involved in antibody response to Mycobacterium avium ssp. paratuberculosis in cattle.

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    Giulietta Minozzi

    Full Text Available BACKGROUND: Mycobacterium avium subsp. paratuberculosis (MAP causes chronic enteritis in a wide range of animal species. In cattle, MAP causes a chronic disease called Johne's disease, or paratuberculosis, that is not treatable and the efficacy of vaccine control is controversial. The clinical phase of the disease is characterised by diarrhoea, weight loss, drop in milk production and eventually death. Susceptibility to MAP infection is heritable with heritability estimates ranging from 0.06 to 0.10. There have been several studies over the last few years that have identified genetic loci putatively associated with MAP susceptibility, however, with the availability of genome-wide high density SNP maker panels it is now possible to carry out association studies that have higher precision. METHODOLOGY/PRINCIPAL FINDINGS: The objective of the current study was to localize genes having an impact on Johne's disease susceptibility using the latest bovine genome information and a high density SNP panel (Illumina BovineSNP50 BeadChip to perform a case/control, genome-wide association analysis. Samples from MAP case and negative controls were selected from field samples collected in 2007 and 2008 in the province of Lombardy, Italy. Cases were defined as animals serologically positive for MAP by ELISA. In total 966 samples were genotyped: 483 MAP ELISA positive and 483 ELISA negative. Samples were selected randomly among those collected from 119 farms which had at least one positive animal. CONCLUSION/SIGNIFICANCE: THE ANALYSIS OF THE GENOTYPE DATA IDENTIFIED SEVERAL CHROMOSOMAL REGIONS ASSOCIATED WITH DISEASE STATUS: a region on chromosome 12 with high significance (P<5x10(-6, while regions on chromosome 9, 11, and 12 had moderate significance (P<5x10(-5. These results provide evidence for genetic loci involved in the humoral response to MAP. Knowledge of genetic variations related to susceptibility will facilitate the incorporation of this information

  9. Intestinal infection following aerosol challenge of calves with Mycobacterium avium subspecies paratuberculosis

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    Eisenberg Susanne WF

    2011-12-01

    Full Text Available Abstract A challenge experiment was performed to investigate whether administration of Mycobacterium avium subsp. paratuberculosis (MAP via the respiratory route leads to MAP infection in calves. Eighteen calves from test negative dams were randomly allocated to four groups. Six calves were challenged with MAP nasally and six calves were challenged by transtracheal injection; three orally challenged calves served as positive controls, and three non challenged calves as negative controls. The challenge was performed as a nine-fold trickle dose, 107 CFU in total. Blood and faecal samples were collected frequently. Calves were euthanized three months post-challenge and extensively sampled. Blood samples were tested for the presence of antibodies and interferon gamma producing cells by ELISA. Faecal and tissue samples were cultured in a liquid culture system and the presence of MAP was confirmed by IS900 realtime PCR. Fourteen out of fifteen calves had no MAP antibody response. The negative controls remained negative; all positive controls became infected. Two nasally challenged calves showed a Purified Protein Derivative Avian (PPDA specific interferon gamma response. In all nasally challenged calves, MAP positive intestinal samples were detected. In three calves of the nasal group MAP positive retropharyngeal lymph nodes or tonsils were detected. In all calves of the transtracheal group MAP positive intestinal tissues were detected as well and three had a MAP positive tracheobronchial lymph node. These findings indicate that inhalation of MAP aerosols can result in infection. These experimental results may be relevant for transmission under field conditions since viable MAP has been detected in dust on commercial dairy farms.

  10. Mycobacterium avium subspecies paratuberculosis infection modifies gut microbiota under different dietary conditions in a rabbit model

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    Rakel eArrazuria

    2016-03-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (MAP the causative agent of paratuberculosis, produces a chronic granulomatous inflammation of the gastrointestinal tract of ruminants. It has been recently suggested that MAP infection may be associated with dysbiosis of intestinal microbiota in ruminants. Since diet is one of the key factors affecting the balance of microbial populations in the digestive tract, we intended to evaluate the effect of MAP infection in a rabbit model fed a regular or high fiber diet during challenge. The composition of microbiota of the cecal content and the sacculus rotundus was studied in 20 New Zealand white female rabbits. The extracted DNA was subjected to paired-end Illumina sequencing of the V3-V4 hypervariable region of the 16S rRNA gene for microbiota analysis. Microbial richness (Chao1 in the cecal content was significantly increased by MAP infection in regular diet rabbits (p = 0.0043 and marginally increased (p = 0.0503 in the high fiber group. Analysis of beta-diversity showed that MAP infection produces deeper changes in the microbiota of sacculus rotundus than in the cecal content. A lower abundance of Proteobacteria in the cecal content of infected animals fed the high fiber diet and also lower abundance of Bacteroidetes in the sacculus rotundus of infected animals fed the regular diet were observed. Based on OPLS-DA analysis, we observed that some bacteria repeatedly appear positively associated with infection in different samples under different diets (families Dehalobacteriaceae, Coriobacteriaceae and Mogibacteriaceae; genus Anaerofustis. The same phenomenon was observed with some of the bacteria negatively associated with MAP infection (genera Anaerostipes and Coprobacillus. However, other groups of bacteria (Enterobacteriaceae family and ML615J-28 order were positively associated with infection in some circumstances and negatively associated with infection in others.Data demonstrate that MAP

  11. Mycobacterium avium complex augments macrophage HIV-1 production and increases CCR5 expression.

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    Wahl, S M; Greenwell-Wild, T; Peng, G; Hale-Donze, H; Doherty, T M; Mizel, D; Orenstein, J M

    1998-10-13

    Infection with HIV-1 results in pronounced immune suppression and susceptibility to opportunistic infections (OI). Reciprocally, OI augment HIV-1 replication. As we have shown for Mycobacterium avium complex (MAC) and Pneumocystis carinii, macrophages infected with opportunistic pathogens and within lymphoid tissues containing OI, exhibit striking levels of viral replication. To explore potential underlying mechanisms for increased HIV-1 replication associated with coinfection, blood monocytes were exposed to MAC antigens (MAg) or viable MAC and their levels of tumor necrosis factor alpha (TNFalpha) and HIV-1 coreceptors monitored. MAC enhanced TNFalpha production in vitro, consistent with its expression in coinfected lymph nodes. Using a polyclonal antibody to the CCR5 coreceptor that mediates viral entry of macrophage tropic HIV-1, a subset of unstimulated monocytes was shown to be CCR5-positive by fluorescence-activated cell sorter analysis. After stimulation with MAg or infection with MAC, CCR5 expression was increased at both the mRNA level and on the cell surface. Up-regulation of CCR5 by MAC was not paralleled by an increase in the T cell tropic coreceptor, CXCR4. Increases in NF-kappaB, TNFalpha, and CCR5 were consistent with the enhanced production of HIV-1 in MAg-treated adherent macrophage cultures as measured by HIV-1 p24 levels. Increased CCR5 was also detected in coinfected lymph nodes as compared with tissues with only HIV-1. The increased production of TNFalpha, together with elevated expression of CCR5, provide potential mechanisms for enhanced infection and replication of HIV-1 by macrophages in OI-infected cells and tissues. Consequently, treating OI may inhibit not only the OI-induced pathology, but also limit the viral burden.

  12. Metabolomic profiling in cattle experimentally infected with Mycobacterium avium subsp. paratuberculosis.

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    Jeroen De Buck

    Full Text Available The sensitivity of current diagnostics for Johne's disease, a slow, progressing enteritis in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP, is too low to reliably detect all infected animals in the subclinical stage. The objective was to identify individual metabolites or metabolite profiles that could be used as biomarkers of early MAP infection in ruminants. In a monthly follow-up for 17 months, calves infected at 2 weeks of age were compared with aged-matched controls. Sera from all animals were analyzed by 1H nuclear magnetic resonance spectrometry. Spectra were acquired, processed, and quantified for analysis. The concentration of many metabolites changed over time in all calves, but some metabolites only changed over time in either infected or non-infected groups and the change in others was impacted by the infection. Hierarchical multivariate statistical analysis achieved best separation between groups between 300 and 400 days after infection. Therefore, a cross-sectional comparison between 1-year-old calves experimentally infected at various ages with either a high- or a low-dose and age-matched non-infected controls was performed. Orthogonal Projection to Latent Structures Discriminant Analysis (OPLS DA yielded distinct separation of non-infected from infected cattle, regardless of dose and time (3, 6, 9 or 12 months after infection. Receiver Operating Curves demonstrated that constructed models were high quality. Increased isobutyrate in the infected cattle was the most important agreement between the longitudinal and cross-sectional analysis. In general, high- and low-dose cattle responded similarly to infection. Differences in acetone, citrate, glycerol and iso-butyrate concentrations indicated energy shortages and increased fat metabolism in infected cattle, whereas changes in urea and several amino acids (AA, including the branched chain AA, indicated increased protein turnover. In conclusion, metabolomics

  13. Metabolic adaptation of Mycobacterium avium subsp. paratuberculosis to the gut environment.

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    Weigoldt, Mathias; Meens, Jochen; Bange, Franz-Christoph; Pich, Andreas; Gerlach, Gerald F; Goethe, Ralph

    2013-02-01

    Knowledge on the proteome level about the adaptation of pathogenic mycobacteria to the environment in their natural hosts is limited. Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne's disease, a chronic and incurable granulomatous enteritis of ruminants, and has been suggested to be a putative aetiological agent of Crohn's disease in humans. Using a comprehensive LC-MS-MS and 2D difference gel electrophoresis (DIGE) approach, we compared the protein profiles of clinical strains of MAP prepared from the gastrointestinal tract of diseased cows with the protein profiles of the same strains after they were grown in vitro. LC-MS-MS analyses revealed that the principal enzymes for the central carbon metabolic pathways, including glycolysis, gluconeogenesis, the tricaboxylic acid cycle and the pentose phosphate pathway, were present under both conditions. Moreover, a broad spectrum of enzymes for β-oxidation of lipids, nine of which have been shown to be necessary for mycobacterial growth on cholesterol, were detected in vivo and in vitro. Using 2D-DIGE we found increased levels of several key enzymes that indicated adaptation of MAP to the host. Among these, FadE5, FadE25 and AdhB indicated that cholesterol is used as a carbon source in the bovine intestinal mucosa; the respiratory enzymes AtpA, NuoG and SdhA suggested increased respiration during infection. Furthermore higher levels of the pentose phosphate pathway enzymes Gnd2, Zwf and Tal as well as of KatG, SodA and GroEL indicated a vigorous stress response of MAP in vivo. In conclusion, our results provide novel insights into the metabolic adaptation of a pathogenic mycobacterium in its natural host.

  14. Systemic and mucosal immune reactivity upon Mycobacterium avium ssp. paratuberculosis infection in mice.

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    Koc, Arzu; Bargen, Imke; Suwandi, Abdulhadi; Roderfeld, Martin; Tschuschner, Annette; Rath, Timo; Gerlach, Gerald F; Hornef, Mathias; Goethe, Ralph; Weiss, Siegfried; Roeb, Elke

    2014-01-01

    Mycobacterium avium ssp. paratuberculosis (MAP) is the cause of Johne's disease, an inflammatory bowel disorder of ruminants. Due to the similar pathology, MAP was also suggested to cause Crohn's disease (CD). Despite of intensive research, this question is still not settled, possibly due to the lack of versatile mouse models. The aim of this study was to identify basic immunologic mechanisms in response to MAP infection. Immune compromised C57BL/6 Rag2-/- mice were infected with MAP intraperitoneally. Such chronically infected mice were then reconstituted with CD4+ and CD8+ T cells 28 days after infection. A systemic inflammatory response, detected as enlargement of the spleen and granuloma formation in the liver, was observed in mice infected and reconstituted with CD4+ T cells. Whereby inflammation in infected and CD4+CD45RB(hi) T cell reconstituted animals was always higher than in the other groups. Reconstitution of infected animals with CD8+ T cells did not result in any inflammatory signs. Interestingly, various markers of inflammation were strongly up-regulated in the colon of infected mice reconstituted with CD4+CD45RB(lo/int) T cells. We propose, the usual non-colitogenic CD4+CD45RB(lo/int) T cells were converted into inflammatory T cells by the interaction with MAP. However, the power of such cells might be not sufficient for a fully established inflammatory response in the colon. Nevertheless, our model system appears to mirror aspects of an inflammatory bowel disease (IBD) like CD and Johne's diseases. Thus, it will provide an experimental platform on which further knowledge on IBD and the involvement of MAP in the induction of CD could be acquired.

  15. Systemic and mucosal immune reactivity upon Mycobacterium avium ssp. paratuberculosis infection in mice.

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    Arzu Koc

    Full Text Available Mycobacterium avium ssp. paratuberculosis (MAP is the cause of Johne's disease, an inflammatory bowel disorder of ruminants. Due to the similar pathology, MAP was also suggested to cause Crohn's disease (CD. Despite of intensive research, this question is still not settled, possibly due to the lack of versatile mouse models. The aim of this study was to identify basic immunologic mechanisms in response to MAP infection. Immune compromised C57BL/6 Rag2-/- mice were infected with MAP intraperitoneally. Such chronically infected mice were then reconstituted with CD4+ and CD8+ T cells 28 days after infection. A systemic inflammatory response, detected as enlargement of the spleen and granuloma formation in the liver, was observed in mice infected and reconstituted with CD4+ T cells. Whereby inflammation in infected and CD4+CD45RB(hi T cell reconstituted animals was always higher than in the other groups. Reconstitution of infected animals with CD8+ T cells did not result in any inflammatory signs. Interestingly, various markers of inflammation were strongly up-regulated in the colon of infected mice reconstituted with CD4+CD45RB(lo/int T cells. We propose, the usual non-colitogenic CD4+CD45RB(lo/int T cells were converted into inflammatory T cells by the interaction with MAP. However, the power of such cells might be not sufficient for a fully established inflammatory response in the colon. Nevertheless, our model system appears to mirror aspects of an inflammatory bowel disease (IBD like CD and Johne's diseases. Thus, it will provide an experimental platform on which further knowledge on IBD and the involvement of MAP in the induction of CD could be acquired.

  16. A Mycobacterium avium subsp. paratuberculosis predicted serine protease is associated with acid stress and intraphagosomal survival

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    Abirami Kugadas

    2016-08-01

    Full Text Available AbstractThe ability to maintain intra-cellular pH is crucial for bacteria and other microbes to survive in diverse environments, particularly those that undergo fluctuations in pH. Mechanisms of acid resistance remain poorly understood in mycobacteria. Although studies investigating acid stress in M. tuberculosis are gaining traction, few center on Mycobacterium avium subsp. paratuberculosis (MAP, the etiological agent of chronic enteritis in ruminants. We identified a MAP acid stress response network involved in macrophage infection. The central node of this network was MAP0403, a predicted serine protease that shared an 86% amino acid identity with MarP in M. tuberculosis. Previous studies confirmed MarP as a serine protease integral to maintaining intra-bacterial pH and survival in acid in vitro and in vivo. We show that MAP0403 is upregulated in infected macrophage and MAC-T cells and coincided with phagosome acidification. Treatment of mammalian cells with bafilomcyin A1, a potent inhibitor of phagosomal vATPases, diminished MAP0403 transcription. MAP0403 expression was also noted in acidic medium. A surrogate host, M. smegmatis mc2 155, was designed to express MAP0403 and when exposed to either macrophages or in vitro acid stress had increase bacterial cell viability, which corresponds to maintenance of intra-bacterial pH in acidic (pH = 5 conditions. These data suggest that MAP0403 may be the equivalent of MarP in MAP. Future studies confirming MAP0403 as a serine protease and exploring its structure and possible substrates are warranted.

  17. Generation and screening of a comprehensive Mycobacterium avium subsp. paratuberculosis transposon mutant bank

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    Govardhan eRathnaiah

    2014-10-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (MAP is the etiologic agent of Johne’s Disease in ruminants. This enteritis has significant economic impact and worldwide distribution. Vaccination is one of the most cost effective infectious disease control measures. Unfortunately, current vaccines reduce clinical disease and shedding, but are of limited efficacy and do not provide long-term protective immunity. Several strategies have been followed to mine the MAP genome for virulence determinants that could be applied to vaccine and diagnostic assay development. In this study, a comprehensive mutant bank of 13,536 MAP K-10 Tn5367 mutants (P > 95% was constructed and screened in vitro for phenotypes related to virulence. This strategy was designated to maximize identification of genes important to MAP pathogenesis without relying on studies of other mycobacterial species that may not translate into similar effects in MAP. This bank was screened for mutants with colony morphology alterations, susceptibility to D-cycloserine, impairment in siderophore production or secretion, reduced cell association, and decreased biofilm and clump formation. Mutants with interesting phenotypes were analyzed by PCR, Southern blotting and DNA sequencing to determine transposon insertion sites. These insertion sites mapped upstream from the MAP1152-MAP1156 cluster, internal to either the Mod operon gene MAP1566 or within the coding sequence of lsr2, and several intergenic regions. Growth curves in broth cultures, invasion assays and kinetics of survival and replication in primary bovine macrophages were also determined. The ability of vectors carrying Tn5370 to generate stable MAP mutants was also investigated.

  18. Preparation and Purification of Polyclonal Antibodies against Mycobacterium Avium Paratuberculosis Antigens in Rabbit

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    Hafezeh Alizadeh

    2012-12-01

    Full Text Available Background and Objective: Johne’s disease is the chronic granulomatous enteritis of ruminants, and a major health hazard worldwide. In recent years, researchers have focused on mycobacterium avium subsp. paratuberculosis (MAP antigens in diagnostic tests. Identification of antibodies against MAP antigens is, therefore, effective for the diagnosis or preparation of vaccine. The aim of this study was to prepare and purify polyclonal antibodies against MAP antigens. Materials and Methods: A New Zealand white rabbit was immunized at a certain time period with MAP antigens and Freund’s adjuvant. After the immunization of the animal, the rabbit was bled to obtain enriched serum. Immunoglobulins were obtained via sedimentation with ammonium sulfate 35% and then IgG was purified by ion exchange (DEAE-cellulose chromatography. Serologic test was used to evaluate the interaction of antigens and antibodies. Results: Ion exchange chromatography of IgG showed one peak, and SDS_PAGE of IgG showed a single band. Serologic test was applied and clear precipitation lines were appeared up to 1:16 dilution, which indicated the high quality of the product. Conclusion: In this study, the humoral immune response was induced well by immunization with MAP antigens in a New Zealand white rabbit and polyclonal antibodies were produced in high titers. Polyclonal antibodies are relatively inexpensive and easy to produce in large quantities and can connect to the more connective sites, resulting in better sensitivity. Identification of polyclonal antibodies via immunological tests can play a significant role in studying MAP disorders.

  19. A Rational Framework for Evaluating the Next Generation of Vaccines Against Mycobacterium avium subspecies paratuberculosis

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    John eBannantine

    2014-09-01

    Full Text Available Since the early 1980s, several investigations have focused on developing a vaccine against Mycobacterium avium subspecies paratuberculosis (MAP, the causative agent of Johne’s disease in cattle and sheep. These studies used whole-cell inactivated vaccines that have proven useful in limiting disease progression, but have not prevented infection. In contrast, modified live vaccines that invoke a Th1 type immune response, may improve protection against infection. Spurred by recent advances in the ability to create defined knockouts in MAP, several independent laboratories have developed modified live vaccine candidates by transpositional mutation of virulence and metabolic genes in MAP. In order to accelerate the process of identification and comparative evaluation of the most promising modified live MAP vaccine candidates, members of a multi-institutional USDA-funded research consortium, the Johne’s disease integrated program (JDIP, met to establish a standardized testing platform using agreed upon protocols. A total of 22 candidates vaccine strains developed in five independent laboratories in the United States and New Zealand voluntarily entered into a double blind stage gated trial pipeline. In Phase I, the survival characteristics of each candidate were determined in bovine macrophages. Attenuated strains moved to Phase II, where tissue colonization of C57/BL6 mice were evaluated in a challenge model. In Phase III, five promising candidates from Phase I and II were evaluated for their ability to reduce fecal shedding, tissue colonization and pathology in a baby goat challenge model. Formation of a multi-institutional consortium for vaccine strain evaluation has revealed insights for the implementation of vaccine trials for Johne’s disease and other animal pathogens. We conclude by suggesting the best way forward based on this 3-phase trial experience and challenge the rationale for use of a macrophage-to-mouse-to native host pipeline for

  20. A rational framework for evaluating the next generation of vaccines against Mycobacterium avium subspecies paratuberculosis.

    Science.gov (United States)

    Bannantine, John P; Hines, Murray E; Bermudez, Luiz E; Talaat, Adel M; Sreevatsan, Srinand; Stabel, Judith R; Chang, Yung-Fu; Coussens, Paul M; Barletta, Raúl G; Davis, William C; Collins, Desmond M; Gröhn, Yrjö T; Kapur, Vivek

    2014-01-01

    Since the early 1980s, several investigations have focused on developing a vaccine against Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne's disease in cattle and sheep. These studies used whole-cell inactivated vaccines that have proven useful in limiting disease progression, but have not prevented infection. In contrast, modified live vaccines that invoke a Th1 type immune response, may improve protection against infection. Spurred by recent advances in the ability to create defined knockouts in MAP, several independent laboratories have developed modified live vaccine candidates by transpositional mutation of virulence and metabolic genes in MAP. In order to accelerate the process of identification and comparative evaluation of the most promising modified live MAP vaccine candidates, members of a multi-institutional USDA-funded research consortium, the Johne's disease integrated program (JDIP), met to establish a standardized testing platform using agreed upon protocols. A total of 22 candidates vaccine strains developed in five independent laboratories in the United States and New Zealand voluntarily entered into a double blind stage gated trial pipeline. In Phase I, the survival characteristics of each candidate were determined in bovine macrophages. Attenuated strains moved to Phase II, where tissue colonization of C57/BL6 mice were evaluated in a challenge model. In Phase III, five promising candidates from Phase I and II were evaluated for their ability to reduce fecal shedding, tissue colonization and pathology in a baby goat challenge model. Formation of a multi-institutional consortium for vaccine strain evaluation has revealed insights for the implementation of vaccine trials for Johne's disease and other animal pathogens. We conclude by suggesting the best way forward based on this 3-phase trial experience and challenge the rationale for use of a macrophage-to-mouse-to native host pipeline for MAP vaccine development.

  1. Evaluation of immunodominant proteins of Mycobacterium avium paratuberculosis cell wall by Western blot analysis.

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    Hashemi, Maryam; Madani, Rasool; Razmi, Nematollah

    2014-04-01

    Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is a slow growing mycobactin, whose dependence on mycobacterial species is known to be the causative agent of Johne's disease (paratuberculosis) in all species of domestic ruminants worldwide. The organism is transmitted via close contact, ingestion, or transplacentally from mother to fetus and occurs commonly in grazing domestic animals. Johne's disease (JD) is characterized by gradual weight loss, decreased milk production, and diarrhea due to the chronic, progressive, granulomatous enteritis and lymphadenitis. The disease can cause serious economic damage to the dairy industry due to the loss of milk production and early culling of infected animals. In recent years, researchers have focused on the identification of a specific antigen of M. paratuberculosis to use in diagnosis test and preparation of effective vaccine. The goal of this study is evaluation of the immunodominant proteins of M. paratuberculosis cell wall. The amount of protein was determined with a Lowry assay (22.68 μg/100 μL). For production of polyclonal antibody against proteins of M. paratuberculosis cell wall, a New Zealand white rabbit was immunized with antigen and Freund's adjuvant. After immunization, the rabbit was bled to produce enriched serum. Antibodies were purified from serum with ion exchange chromatography. In the Ouchterlony test, the reactions between antigen and antibodies were seen in dilutions of one quarter for serum, one quarter for Ig, and one half for IgG by clear precipitation lines due to the well immunization of the rabbit. Electrophoresis and Western blot analysis were used and subsequently a sharp band appeared in nitrocellulose paper; these bands were about 25, 37, 50, 75, and 150 kDa molecular weight, which indicated immunodominant proteins.

  2. Sardinian Type 1 diabetes patients, Transthyretin and Mycobacterium avium subspecies paratuberculosis infection

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    Masala Speranza

    2012-12-01

    Full Text Available Abstract Background Mycobacterium avium subsp. paratuberculosis (MAP is the cause of Johne’s disease, an enteric granulomatous disease. Recently, MAP has been associated with different autoimmune diseases such as Crohn’s disease, type 1 diabetes (T1D and multiple sclerosis. Transthyretin (TTR is a plasma transport protein for thyroid hormone and forms a complex with retinol-binding protein. Reduced TTR plasma levels in MAP infected ovines have been reported. TTR exerts also a functional role in the pancreas promoting insulin release and protecting β-cells from death. Our objective was to identify a protein that could be used as a diagnostic marker of T1D for determining disease progression and monitoring at-risk patients. We postulate that serological TTR levels would be reduced in T1D MAP exposed patients. Our hypothesis is based on the observation of cases of T1D patients with decreased TTR levels beside the reduced TTR plasma levels in ovines with Johne’s disease. We quantified the plasma protein levels of TTR in 50 people with T1D and 51 age-matched healthy controls (HCs by means of enzyme-linked immunosorbent assays (ELISA. Findings Our pilot study showed that plasma TTR levels were not significantly lower/higher in T1D Sardinian cases compared to the HCs. Conclusion These preliminary data indicate that plasma TTR may not be a good candidate biomarker for T1D diagnosis and further studies to elucidate the possible link are needed.

  3. The possible involvement of Mycobacterium avium ssp. paratuberculosis in the aetiology of Crohn's disease: a case control study in the Netherlands

    NARCIS (Netherlands)

    Herrewegh AAPM; Overduin P; Roholl PJM; Gielis FK; Robinson JE; Mahmmod N; Lieverse RJ; Robijn RJ; Zanden AGM van der; Soolingen D van; Dept of Medical Microbiology; Biomedics Scientific Consultancy; LIS; LPI

    2005-01-01

    A case control study was performed to investigate the possible role of Mycobacterium avium ssp. paratuberculosis (Map) in the aetiology of Crohn's disease (CD). Biopsy samples were collected from the ileum and colon of CD patients, Ulcerative Colitis (UC) patients and control persons. The biopsy sam

  4. The possible involvement of Mycobacterium avium ssp. paratuberculosis in the aetiology of Crohn's disease: a case control study in the Netherlands

    NARCIS (Netherlands)

    Herrewegh AAPM; Overduin P; Roholl PJM; Gielis FK; Robinson JE; Mahmmod N; Lieverse RJ; Robijn RJ; van der Zanden AGM; van Soolingen D; LIS; LPI

    2005-01-01

    De bacterie Mycobacterium avium ssp. paratuberculosis (Map) wordt beschouwd als een mogelijke oorzaak van de ziekte van Crohn (morbus Crohn, MC). In samenwerking met Gelre ziekenhuizen heeft het RIVM een onderzoek uitgevoerd naar het voorkomen van Map in darmbiopten van patienten met MC, patienten

  5. A longitudinal study of factors influencing the result of a Mycobacterium avium ssp. paratuberculosis antibody ELISA in milk or dairy cows

    NARCIS (Netherlands)

    Eisenberg, S.W.F.; Veldman, E.; Rutten, V.P.M.G.; Koets, A.P.

    2015-01-01

    The influence of milk yield and milk composition on the diagnosis of Mycobacterium avium ssp. paratuberculosis (MAP) by milk ELISA in the context of the total IgG secretion patterns in milk throughout lactation and serum concentrations were investigated. A 2-yr trial was performed in which 1,410 dai

  6. Susceptibility to paratuberculosis infection in cattle is associated withsingle nucleotide polymorphisms in Toll-like receptor 2 which modulate immune responses against Mycobacterium avium subspecies paratuberculosis

    DEFF Research Database (Denmark)

    Koets, A; Santema, W; Oostenriik, D;

    2010-01-01

    Paratuberculosis is a chronic intestinal infection in ruminants, caused by Mycobacterium avium subspecies paratuberculosis (Map). To study the role of host genetics in disease susceptibility, the Toll-like receptor 2 (TLR2) gene, selected based on its potential role in immunity to mycobacterial i...

  7. Mycobacterium avium subsp. paratuberculosis PPE Protein MAP1152 and Conserved Protein MAP1156 are Antigenic in Experimentally and Naturally Infected Cattle

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne’s Disease (JD) in ruminants resulting in significant production losses. An insertion mutation upstream from the MAP1152-MAP1156 region causes a change in colony morphotype and results in an attenuated phenotype in bovine monocyte derive...

  8. Optimization of methods for the detection of Mycobacterium avium subsp. paratuberculosis in milk and colostrum of naturally infected dairy cows with Johne's Disease

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s Disease (JD), a chronic enteritis that occurs in dairy cattle and other ruminants. A 2007 NAHMS Dairy Study demonstrated that over 68% of dairy herds are infected with JD so the risk of exposure within a herd is high...

  9. Effect of feeding heat-treated colostrum on risk for infection with Mycobacterium avium subsp. paratuberculosis, milk production and longevity in Holstein dairy cows

    Science.gov (United States)

    In summer 2007, a randomized controlled clinical trial was initiated on 6 large Midwest commercial dairy farms to investigate the effect of feeding heat-treated (HT) colostrum on transmission of Mycobacterium avium subsp. paratuberculosis and on future milk production and longevity within the herd. ...

  10. Evaluation of the association between fecal excretion of Mycobacterium avium subsp paratuberculosis and detection in colostrum and on teat skin surfaces of dairy cows

    Science.gov (United States)

    Objective—To evaluate the association between fecal excretion of Mycobacterium avium subsp paratuberculosis (MAP) by dairy cows in the periparturient period and detection of MAP DNA in colostrum specimens and on teat skin surfaces. Design—Cross-sectional study. Animals—112 Holstein cows. Procedures—...

  11. Clinical disease and stage of lactation influences shedding of Mycobacterium avium subsp. paratuberculosis into milk and colostrum of naturally infected dairy cows

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease (JD). One mode of transmission of MAP is through ingestion of contaminated milk and colostrum by susceptible calves. The objective of this study was to determine if the amount of MAP shed into the milk and co...

  12. Dam Mycobacterium avium subspecies parartuberculosis (MAP) infection status does not predetermine calves for future shedding when raised in a contaminated environment

    NARCIS (Netherlands)

    Eisenberg, S.W.F.; Rutten, Victor P.M.G.; Koets, A.P.

    2015-01-01

    Uptake of Mycobacterium avium subsp. paratuberculosis (MAP) by calves in the first days of life from colostrum, milk and faeces is regarded an important moment of transmission. The objective of this study was to quantify the
    association between the MAP status of dams as determined by the presenc

  13. Predicting the role of IL-10 in the regulation of the adaptive immune responses in Mycobacterium avium subsp. paratuberculosis infections using mathematical models

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is an intracellular bacterial pathogen that causes Johne’s disease in cattle and other animals. Infection follows ingestion of the bacteria primarily through the fecal oral route and results in the colonization of the intestine and a granulomatous en...

  14. Infections with Mycobacterium tuberculosis and Mycobacterium avium among HIV-infected patients after the introduction of highly active antiretroviral therapy. EuroSIDA Study Group JD

    DEFF Research Database (Denmark)

    Kirk, O; Gatell, J M; Mocroft, A;

    2000-01-01

    the introduction of HAART, using data from the EuroSIDA study, a European, multicenter observational cohort of more than 7,000 patients. Overall incidences of Mycobacterium tuberculosis (TB) and Mycobacterium avium complex (MAC) were 0.8 and 1.4 cases/100 person-years of follow-up (PYF), decreasing from 1.8 (TB...

  15. Apparent prevalence of beef carcasses contaminated with Mycobacterium avium subsp. paratuberculosis sampled from Danish slaughter cattle

    DEFF Research Database (Denmark)

    Okura, Hisako; Toft, Nils; Pozzato, Nicola;

    2011-01-01

    Presence of Mycobacterium avium subsp. paratuberculosis (MAP) in beef has been reported as a public health concern because asymptomatically infected cattle may contain MAP in tissues that are used for human consumption. Associations between MAP carcasses contamination and animal characteristics s...

  16. Epidemiological characterization and risk factors associated with Mycobacterium avium subsp. paratuberculosis infection in dairy goats in the Brazilian semiarid region

    Directory of Open Access Journals (Sweden)

    Theonys Diógenes Freitas

    2015-02-01

    Full Text Available The aim of this investigation was to conduct an epidemiological study and identify risk factors associated with the occurrence of paratuberculosis (Johne’s disease in dairy goats within the semiarid region of Paraíba State. The study was done during the period of March 2009 to July 2011, during which 727 female goats from 86 flocks from the city of Monteiro, Paraíba were investigated. For the serological diagnosis of Mycobacterium avium subsp. paratuberculosis (Map infection indirect ELISA tests (screening and confirmatory were performed. Of the 727 animals used six (0.82% were seropositive at the confirmatory test after screening, and of the 86 flocks six (6.97% presented at least one seropositive animal. In positive flocks the frequency of reactive animals ranged from 5.26% to 16.60%. Risk factors identified were production system (weaning and reproduction (odds ratio = 36.0; 95% CI = 2.6 –486.1; p < 0,001 and absence of technical infrastructure (odds ratio = 54.0; 95% CI = 4.5 –642.9; p < 0,001. It was concluded that Mycobacterium avium subsp. paratuberculosis is present in dairy goat flocks in the region; however, its influence on decrease productivity as well as the risk of transmission to humans through animal products must totally evaluated. Based on the analysis of risk factors, improvements are recommended for the technical infrastructure and the management of breeding goats.

  17. Molecular and quantitative signatures of biparental inbreeding depression in the self-incompatible tree species Prunus avium.

    Science.gov (United States)

    Jolivet, C; Rogge, M; Degen, B

    2013-05-01

    Genetic diversity strongly influences populations' adaptability to changing environments and therefore survival. Sustainable forest management practices have multiple roles including conservation of genetic resources and timber production. In this study, we aimed at better understanding the variation in genetic diversity among adult and offspring individuals, and the effects of mating system on offspring survival and growth in wild cherry, Prunus avium. We analysed adult trees and open pollinated seed-families from three stands in Germany at eight microsatellite loci and one incompatibility system locus and conducted paternity analyses. Seed viability testing and seed sowing in a nursery allowed further testing for the effects of pollen donor diversity and genetic similarity between mates on the offspring performance at the seed and seedling stages. Our results were contrasting across stands. Loss of genetic diversity from adult to seedling stages and positive effect of mate diversity on offspring performance occurred in one stand only, whereas biparental inbreeding depression and significant decrease in fixation index from adults to seedlings was detected in two stands. We discussed the effects of stand genetic diversity on the magnitude of biparental inbreeding depression at several life-stages and its consequences on the management of genetic resources in P. avium.

  18. Genomic Characterization of the Vaccinal Strain of Mycobacterium Avium Subspecies Paratuberculosis (MAP 316F by MIRU-VNTR

    Directory of Open Access Journals (Sweden)

    Zahra Ebrahim (MSc

    2015-10-01

    Full Text Available Background and Objective: Paratuberculosis is a chronic granulomatous enteritis of ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP. this study aimed to characterize the genome of the MAP 316F strain. Methods: The MAP 316F strain was subjected to the PCR-F57 and PCR-IS900 experiments in order to ensure its identity as MAP. This was followed by application of the Thibault genotyping system consisting of eight loci including 292, x3, 25, 47, 3, 7, 10 and 32. Required genomic material for all experiments was prepared using the simple method of boiling. Gel electrophoresis findings related to the typing PCRs were backed by sequencing of amplification products. Results: In PCR amplification, eight products with the size of 300, 298, 350, 217, 208, 203, 803 and 649 bp were detected at 292, X3, 25, 47, 3, 7, 10 and 32 loci, holding 3, 2, 3, 3, 2, 2, 2 and 8 copies of TRs at these loci, respectively. Conclusion: This genomic pattern is matched with that of the MAP 316F vaccine strain from the French Merial company and also the MAP K10 fully-sequenced strain. Keywords: Mycobacterium avium subsp. paratuberculosis, Genomics, Genotyping techniques, Strain

  19. Acidic methanolysis v. alkaline saponification in gas chromatographic characterization of mycobacteria: differentiation between Mycobacterium avium-intracellulare and Mycobacterium gastri.

    Science.gov (United States)

    Larsson, L

    1983-08-01

    Mycobacterium avium-intracellulare and M.gastri were analyzed with capillary gas chromatography after each strain had been subjected to acidic methanolysis or to alkaline saponification followed by methylation. Prominent peaks of myristic, palmitoleic, palmitic, oleic, stearic and tuberculostearic acids were found in the chromatograms of both species, whereas 2-octadecanol and 2-eicosanol were detected only in M. avium-intracellulare. In initial runs, both of the derivatization principles yielded virtually identical chromatograms for a given strain. After repeated injections of extracts from alkaline saponification, however, the alcohol peaks showed pronounced tailing and finally almost disappeared from the chromatograms. This disadvantage, which was not observed when only acid methanolysis was used, could be overcome with trifluoroacetylation. Restored peak shape of the underivatized alcohols could be achieved by washing the cross-linked stationary phase in the capillary tubing with organic solvents. The study demonstrated the importance of conditions which enable separation of 2-octadecanol and 2-eicosanol when gas chromatography is used for species identification of mycobacteria.

  20. Broncho-pleural fistula with hydropneumothorax at CT: Diagnostic implications in mycobacterium avium complex lung disease with pleural involvement

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Hyun Jung; Chung, Myung Jin; Lee, Kyung Soo; Park, Hye Yun; Koh, Won Jung [Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul (Korea, Republic of); Kim, Jung Soo [Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Inha University Hospital, Inha University School of Medicine, Incheon (Korea, Republic of)

    2016-04-15

    To determine the patho-mechanism of pleural effusion or hydropneumothorax in Mycobacterium avium complex (MAC) lung disease through the computed tomographic (CT) findings. We retrospectively collected data from 5 patients who had pleural fluid samples that were culture-positive for MAC between January 2001 and December 2013. The clinical findings were investigated and the radiological findings on chest CT were reviewed by 2 radiologists. The 5 patients were all male with a median age of 77 and all had underlying comorbid conditions. Pleural fluid analysis revealed a wide range of white blood cell counts (410-100690/µL). The causative microorganisms were determined as Mycobacterium avium and Mycobacterium intracellulare in 1 and 4 patients, respectively. Radiologically, the peripheral portion of the involved lung demonstrated fibro-bullous changes or cavitary lesions causing lung destruction, reflecting the chronic, insidious nature of MAC lung disease. All patients had broncho-pleural fistulas (BPFs) and pneumothorax was accompanied with pleural effusion. In patients with underlying MAC lung disease who present with pleural effusion, the presence of BPFs and pleural air on CT imaging are indicative that spread of MAC infection is the cause of the effusion.

  1. ISOLATION, IDENTIFICATION AND BIOINFORMATICS ANALYSIS OF SIX NEWCASTLE DISEASE VIRUSES OF PIGEON ORIGIN%六株鸽源新城疫病毒的分离、鉴定及生物信息学分析

    Institute of Scientific and Technical Information of China (English)

    李仕超; 刘佩红; 丁铲; 仇旭升; 刘开春; 周锦萍; 刘健; 鞠厚斌; 谭磊; 孙英杰; 宋翠萍

    2014-01-01

    showed that four highly virulent strains belonged to ClassⅡⅥb subgenotype and two low virulent strains were similar to La Sota and belonged to ClassⅡgenotypeⅡ. In addition, four highly virulent strains were more closely related to the strain isolated in Italy a few years ago than those strains obtained in China in 1990’s. Therefore, these virulent pigeon strains currently prevailing in China might originate from Europe. Bioinformatics analysis revealed that a linear epitope consisting of amino acids 345-353 on HN protein of these pigeon strains changed as compared with the classic vaccine strain La Sota. In addition, hemagglutination inhibition revealed a noticeable difference between these pigeon strains and La Sota. The results support that chicken vaccine strains do not effectively protect pigeons from NDV infection thus the vaccines designed for pigeon use are much needed.

  2. Streptococcus gallolyticus subsp. gallolyticus from human and animal origins: genetic diversity, antimicrobial susceptibility, and characterization of a vancomycin-resistant calf isolate carrying a vanA-Tn1546-like element.

    Science.gov (United States)

    Romero-Hernández, Beatriz; Tedim, Ana P; Sánchez-Herrero, José Francisco; Librado, Pablo; Rozas, Julio; Muñoz, Gloria; Baquero, Fernando; Cantón, Rafael; Del Campo, Rosa

    2015-04-01

    The aim of this work was to characterize the antibiotic susceptibility and genetic diversity of 41 Streptococcus gallolyticus subsp. gallolyticus isolates: 18 isolates obtained from animals and 23 human clinical isolates. Antibiotic susceptibility was determined by the semiautomatic Wider system and genetic diversity by pulsed-field gel electrophoresis (PFGE) with SmaI. Animal isolates grouped separately in the PFGE analysis, but no statistical differences in antimicrobial resistance were found between the two groups. The LMG 17956 sequence type 28 (ST28) strain recovered from the feces of a calf exhibited high levels of resistance to vancomycin and teicoplanin (MIC, ≥256 mg/liter). Its glycopeptide resistance mechanism was characterized by Southern blot hybridization and a primer-walking strategy, and finally its genome, determined by whole-genome sequencing, was compared with four closely related S. gallolyticus subsp. gallolyticus genomes. Hybridization experiments demonstrated that a Tn1546-like element was integrated into the bacterial chromosome. In agreement with this finding, whole-genome sequencing confirmed a partial deletion of the vanY-vanZ region and partial duplication of the vanH gene. The comparative genomic analyses revealed that the LMG 17956 ST28 strain had acquired an unusually high number of transposable elements and had experienced extensive chromosomal rearrangements, as well as gene gain and loss events. In conclusion, S. gallolyticus subsp. gallolyticus isolates from animals seem to belong to lineages separate from those infecting humans. In addition, we report a glycopeptide-resistant isolate from a calf carrying a Tn1546-like element integrated into its chromosome.

  3. Lactase persistence, NOD2 status and Mycobacterium avium subsp. paratuberculosis infection associations to Inflammatory Bowel Disease

    Directory of Open Access Journals (Sweden)

    Elguezabal Natalia

    2012-06-01

    Full Text Available Abstract Background Inflammatory Bowel Disease (IBD, which includes both Crohn’s disease (CD and ulcerative colitis (UC, is caused by a complex interplay involving genetic predisposition, environmental factors and an infectious agent. Mycobacterium avium subsp. paratuberculosis (MAP is a promising pathogen candidate since it produces a chronic intestinal inflammatory disease in ruminants that resembles CD in humans. MAP is a ubiquitous microorganism, although its presence in the food chain, especially in milk from infected animals, is what made us think that there could be an association between lactase persistence (LP and IBD. The LCT mutation has brought adaptation to dairy farming which in turn would have increased exposure of the population to infection by MAP. NOD2 gene mutations are highly associated to CD. Methods In our study, CD and UC patients and controls from the North of Spain were genotyped for the lactase gene (LCT and for three NOD-2 variants, R702W, G908R and Cins1007fs. MAP PCR was carried out in order to assess MAP infection status and these results were correlated with LCT and NOD2 genotypes. Results As for LP, no association was found with IBD, although UC patients were less likely to present the T/T−13910 variant compared to controls, showing a higher C-allele frequency and a tendency to lactase non-persistence (LNP. NOD2 mutations were associated to CD being the per-allele risk higher for the Cins1007fs variant. MAP infection was more extended among the healthy controls (45.2% compared to CD patients (21.38% and UC patients (19.04% and this was attributed to therapy. The Asturian CD cohort presented higher levels of MAP prevalence (38.6% compared to the Basque CD cohort (15.5%, differences also attributed to therapy. No interaction was found between MAP infection and LCT or NOD2 status. Conclusions We conclude that LP is not significantly associated with IBD, but that MAP infection and NOD2 do show not mutually

  4. The potential Public Health Impact of Mycobacterium avium ssp. paratuberculosis: Global Opinion Survey of Topic Specialists.

    Science.gov (United States)

    Waddell, L A; Rajić, A; Stärk, K D C; McEwen, S A

    2016-05-01

    Global research knowledge has accumulated over the past few decades, and there is reasonable evidence for a positive association between Mycobacterium avium spp. paratuberculosis and Crohn's disease in humans, although its role as a human pathogen has not been entirely accepted. For this reason, management of public health risk due to M. paratuberculosis remains an important policy issue in agri-food public health arenas in many countries. Responsible authorities must decide whether existing mitigation strategies are sufficient to prevent or reduce human exposure to M. paratuberculosis. A Web-based questionnaire was administered to topic specialists to elicit empirical knowledge and opinion on the overall public health impact of M. paratuberculosis, the importance of various routes of human exposure to the pathogen, existing mitigation strategies and the need for future strategies. The questionnaire had four sections and consisted of 20 closed and five open questions. Topic specialists believed that M. paratuberculosis is likely a risk to human health (44.8%) and, given the paucity of available evidence, most frequently ranked it as a moderate public health issue (40.1%). A significant correlation was detected between topic specialists' commitment to M. paratuberculosis in terms of the number of years or proportion of work dedicated to this topic, and the likelihood of an extreme answer (high or low) to the above questions. Topic specialists identified contact with ruminants and dairy products as the most likely routes of exposure for humans. There was consensus on exposure routes for ruminants and what commodities to target in mitigation efforts. Described mandatory programmes mainly focused on culling diseased animals and voluntary on-farm prevention programmes. Despite ongoing difficulties in the identification of subclinical infections in animals, the topic specialists largely agreed that further enhancement of on-farm programmes in affected commodities by

  5. Iron-sparing Response of Mycobacterium avium subsp. paratuberculosis is strain dependent

    Directory of Open Access Journals (Sweden)

    Higgins LeeAnn

    2010-10-01

    Full Text Available Abstract Background Two genotypically and microbiologically distinct strains of Mycobacterium avium subsp. paratuberculosis (MAP exist - S and C MAP strains that primarily infect sheep and cattle, respectively. Concentration of iron in the cultivation medium has been suggested as one contributing factor for the observed microbiologic differences. We recently demonstrated that S strains have defective iron storage systems, leading us to propose that these strains might experience iron toxicity when excess iron is provided in the medium. To test this hypothesis, we carried out transcriptional and proteomic profiling of these MAP strains under iron-replete or -deplete conditions. Results We first complemented M. smegmatisΔideR with IdeR of C MAP or that derived from S MAP and compared their transcription profiles using M. smegmatis mc2155 microarrays. In the presence of iron, sIdeR repressed expression of bfrA and MAP2073c, a ferritin domain containing protein suggesting that transcriptional control of iron storage may be defective in S strain. We next performed transcriptional and proteomic profiling of the two strain types of MAP under iron-deplete and -replete conditions. Under iron-replete conditions, C strain upregulated iron storage (BfrA, virulence associated (Esx-5 and antigen85 complex, and ribosomal proteins. In striking contrast, S strain downregulated these proteins under iron-replete conditions. iTRAQ (isobaric tag for relative and absolute quantitation based protein quantitation resulted in the identification of four unannotated proteins. Two of these were upregulated by a C MAP strain in response to iron supplementation. The iron-sparing response to iron limitation was unique to the C strain as evidenced by repression of non-essential iron utilization enzymes (aconitase and succinate dehydrogenase and upregulation of proteins of essential function (iron transport, [Fe-S] cluster biogenesis and cell division. Conclusions Taken

  6. Occurrence of Mycobacterium avium subspecies paratuberculosis and Neospora caninum in Alberta cow-calf operations.

    Science.gov (United States)

    Pruvot, M; Kutz, S; Barkema, H W; De Buck, J; Orsel, K

    2014-11-01

    Mycobacterium avium subsp. paratuberculosis (MAP) and Neospora caninum (NC) are two pathogens causing important production limiting diseases in the cattle industry. Significant impacts of MAP and NC have been reported on dairy cattle herds, but little is known about the importance, risk factors and transmission patterns in western Canadian cow-calf herds. In this cross-sectional study, the prevalence of MAP and NC infection in southwest Alberta cow-calf herds was estimated, risk factors for NC were identified, and the reproductive impacts of the two pathogens were assessed. Blood and fecal samples were collected from 840 cows on 28 cow-calf operations. Individual cow and herd management information was collected by self-administered questionnaires and one-on-one interviews. Bayesian estimates of the true prevalence of MAP and NC were computed, and bivariable and multivariable statistical analysis were done to assess the association between the NC serological status and ranch management risk factors, and the clinical effects of the two pathogens. Bayesian estimates of true prevalence indicated that 20% (95% probability interval: 8-38%) of herds had at least one MAP-positive cow, with a within-herd prevalence in positive herds of 22% (8-45%). From the Bayesian posterior distributions of NC prevalence, the median herd-level prevalence was 66% (33-95%) with 10% (4-21%) cow-level prevalence in positive herds. Multivariable analysis indicated that introducing purchased animals in the herd might increase the risk of NC. The negative association of NC with proper carcass disposal and presence of horses on ranch (possibly in relation to herd monitoring and guarding activities), may suggest the importance of wild carnivores in the dynamics of this pathogen in the study area. We also observed an association between MAP and NC serological status and the number of abortions. Additional studies should be done to further examine specific risk factors for MAP and NC, assess the

  7. Economic analysis of Mycobacterium avium subspecies paratuberculosis vaccines in dairy herds.

    Science.gov (United States)

    Cho, J; Tauer, L W; Schukken, Y H; Gómez, M I; Smith, R L; Lu, Z; Grohn, Y T

    2012-04-01

    Johne's disease, or paratuberculosis, is a chronic infectious enteric disease of ruminants, caused by infection with Mycobacterium avium ssp. paratuberculosis (MAP). Given the absence of a fail-safe method of prevention or a cure, Johne's disease can inflict significant economic loss on the US dairy industry, with an estimated annual cost of over $200 million. Currently available MAP control strategies include management measures to improve hygiene, culling MAP serologic- or fecal-positive adult cows, and vaccination. Although the 2 first control strategies have been reported to be effective in reducing the incidence of MAP infection, the changes in herd management needed to conduct these control strategies require significant effort on the part of the dairy producer. On the other hand, vaccination is relatively simple to apply and requires minor changes in herd management. Despite these advantages, only 5% of US dairy operations use vaccination to control MAP. This low level of adoption of this technology is due to limited information on its cost-effectiveness and efficacy and some important inherent drawbacks associated with current MAP vaccines. This study investigates the epidemiological effect and economic values of MAP vaccines in various stages of development. We create scenarios for the potential epidemiological effects of MAP vaccines, and then estimate economically justifiable monetary values at which vaccines become economically beneficial to dairy producers such that a net present value (NPV) of a farm's net cash flow can be higher than the NPV of a farm using no control or alternative nonvaccine controls. Any vaccination with either low or high efficacy considered in this study yielded a higher NPV compared with a no MAP control. Moreover, high-efficacy vaccines generated an even higher NPV compared with alternative controls, making vaccination economically attractive. Two high-efficacy vaccines were particularly effective in MAP control and NPV

  8. Environmental contamination with Mycobacterium avium subsp. paratuberculosis in endemically infected dairy herds.

    Science.gov (United States)

    Smith, R L; Schukken, Y H; Pradhan, A K; Smith, J M; Whitlock, R H; Van Kessel, J S; Wolfgang, D R; Grohn, Y T

    2011-10-01

    Environmental contamination with Mycobacterium avium subsp. paratuberculosis (MAP) is thought to be one of the primary sources of infection for dairy cattle. The exact link between fecal shedding of MAP by individual cows and environmental contamination levels at the herd level was explored with a cross-sectional analysis of longitudinally collected samples on 3 dairy farms. Composite samples from multiple environmental sites in 3 commercial dairy herds in the Northeast US were cultured quarterly for MAP, providing 1131 samples (133 (11.8%) were culture-positive), and all adult animals in the herds were tested biannually by fecal culture (FC), for 6 years. Of the environmental sites sampled, manure storage areas and shared alleyways were most likely to be culture-positive. Environmental sample results were compared to FC results from either the concurrent or previous sampling date at both the herd and the pen level. At the herd level, a 1 log unit increase in average fecal shedding increased the odds of a positive non-pen environmental sample by a factor of 6 and increased the average amount of MAP in non-pen samples by 2.9 cfu/g. At the pen level, a 1 log unit increase in average fecal shedding in the pen increased the odds of a positive environment by a factor of 2.4 and the average amount of MAP was increased by 3.5 cfu/g. We were not able to model the relationship between non-pen environmental sample status and the distance between shedding animals and the sample's location, and neighboring pens did not significantly affect the results of the pen-level analysis. The amount of MAP in pen-level samples and the probability of a pen testing positive for MAP were both positively but non-significantly correlated with the number of animals in the pen shedding >30 cfu/g of MAP. At least 6 environmental samples met the criteria for the U.S. Voluntary Bovine Johne's Disease Control Program on 47 of the 72 sampling dates; of these, 19 of the 47 FC-positive sampling dates

  9. Goat milk as a non-invasive sample for confirmation of Mycobacterium avium subspecies paratuberculosis by IS900 PCR

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    Bharathy Sukumar

    2014-09-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (MAP causes Johne's disease (JD in cattle, sheep, goats and other ruminants, and Crohn’s disease in humans. MAPs are shed to external environment through feces and milk. The present study was aimed to evaluate the utility of milk as a non-invasive sample in stage II MAP infections in goats using IS900 polymerase chain reaction (PCR and sequencing analysis. A total of 32 milk samples from lactating does were collected. Within these 32 milk samples, 15 were collected from pre-confirmed JD positive goats. By IS900 PCR, all the 15 (100% known JD positive goat milk samples revealed the presence of MAP. However, no unknown goat was identified as MAP positive. The results of this study established the usefulness of milk as a non-invasive sample in screening and confirmation of stage II MAP infection in goats.

  10. Proteomic Comparison of Fruit Ripening between 'Hedelfinger' Sweet Cherry (Prunus avium L.) and Its Somaclonal Variant 'HS'.

    Science.gov (United States)

    Prinsi, Bhakti; Negri, Alfredo S; Espen, Luca; Piagnani, M Claudia

    2016-05-25

    The somaclonal variant HS, from sweet cherry (Prunus avium L.) 'Hedelfinger' (H), was previously selected for reduced tree vegetative vigor and lesser canopy density. In this work, we compared H and HS fruits at early unripe (green) and full ripe (dark red) stages by biochemical and proteomic approaches. The main biochemical parameters showed that fruit quality was not affected by somaclonal variation. The proteomic analysis identified 39 proteins differentially accumulated between H and HS fruits at the two ripening stages, embracing enzymes involved in several pathways, such as carbon metabolism, cell wall modification, stress response, and secondary metabolism. The evaluation of fruit phenolic composition by mass spectrometry showed that HS sweet cherries have higher levels of procyanidin, flavonol, and anthocyanin compounds. This work provides the first proteomic characterization of fruit ripening in sweet cherry, revealing new positive traits of the HS somaclonal variant.

  11. Effects of Seed Gathering, Sowing Time and Growing Conditions on Wild Cherry (Prunus avium L. Seed Germination

    Directory of Open Access Journals (Sweden)

    Şemsettin Kulaç

    2009-04-01

    Full Text Available Wild cherry (Prunus avium L., in our forests, can be found either as an individual or as in small groups. Hardness and thickness of seed coat, immature embryo, chemical substances in seed and pericarp are well known problems on germination. In this study, Germination impediment tests have done on seeds that were gathered from KTU campus on three different times, on 10th, 20th and 30th of June 2006, these seeds had sown, 200 of each seed with pericarp and with the ones haven’t have pericarp, in four different germination media (soil, perlite, needle and leaf mould to overcome wild cherry’s germination impediment. The highest germination percentage (82.5 % had found among seeds gathered on 20th of June; these were sown in needle mould and had no pericarp. However, our researches show that seeds with pericarp, gathered on 20th of June have maximum germination percentage of (34 % in leaf mould.

  12. Morphological indicators of the quality of one-year-old bare-root seedlings of wild cherry (Prunus avium L.

    Directory of Open Access Journals (Sweden)

    Stjepanović Stefan

    2013-01-01

    Full Text Available This paper presents morphological indicators of the quality of one-year-old seedlings of wild cherry (Prunus avium L. with special reference to their correlations. The examined parameters were: length, diameter, weight of the aboveground and underground parts, number of buds, root volume, root length and leaf area. The ratio of heights was determined on the basis of the parameters derived: diameter, and the ratio of aboveground and underground parts of a seedling and the Quality Index. Height and diameter have proven to be good indicators of quality. Root volume has proven to be a better indicator of the quality of seedlings than root length. The weight ratio of the aboveground and underground parts of broadleaved trees in the dry state must be defined in advance, both when the measurements are carried out with and without leaves. The Quality Index has proven to be the most comprehensive morphological indicator of quality.

  13. Antioxidant Activity and Phenolic Content of Sweet Cherries (Prunus Avium L. from West and South-West of Romania

    Directory of Open Access Journals (Sweden)

    Sofia Popescu

    2014-11-01

    Full Text Available A diet rich in fruits and vegetables is associated with a lower incidence of degenerative diseases (such as cardiovascular disease and certain types of cancers. Currently, most research is focused on the content of polyphenols and antioxidant compounds found in fruit and vegetable. Sweet cherries (Prunus avium L. contain a significant amount of polyphenols and several antioxidants that possess many biological activities such as anticancer, antioxidant and anti-inflammation properties. In present study were investigated the quantification of total polyphenols and antioxidant capacity in fruits of a number of selected sweet cherry genotypes. Although sweet cherry fruits are a significant source of different phenolic compounds, antioxidant activity of sweet cherries is not related only with the total phenolic content.  

  14. Bayesian kriging of seroprevalence to Mycobacterium avium subspecies paratuberculosis and Neospora caninum in Alberta beef and dairy cattle.

    Science.gov (United States)

    Thompson, James A; Scott, H Morgan

    2007-12-01

    Identifying spatial patterns of risk is important in the study of diseases with ecologic causes. Furthermore, relatively complex hierarchical modeling is required to determine how factors that are organized across levels interact, such as how an ecologic cause interacts with farm management and with animal characteristics. The objective of this study was to map the risk for Mycobacterium avium subspecies paratuberculosis (MAP - the causative agent of Johne's disease) and Neospora caninum (NC - the cause of neosporosis) infections in Alberta beef and dairy cattle. This objective utilized Bayesian generalized linear kriging to partition herd effects into a portion attributable to location and a portion that was independent of location. Seropositivity to NC in beef cattle showed strong support for spatial covariance, suggesting that ecologic causes were important for beef cattle but not dairy cattle. There was little evidence of spatial covariance for MAP seropositivity in either beef or dairy cattle.

  15. Multiplex PCR in diagnosis of M. tuberculosis and M. avium co-infection from lymph node in an AIDS patient

    Directory of Open Access Journals (Sweden)

    K Sharma

    2015-01-01

    Full Text Available A 35-year-old, HIV-seropositive male (CD4 count 41 cells/mm 3 on highly active antiretroviral ( HAART presented with fever and weight loss for 3 months and new skin lesions. He was earlier diagnosed of TB and was on anti-tubercular therapy (ATT. The retroperitoneal lymph node aspirate showed acid-fast bacilli and epithelioid cell granulomas; however, cultures remained sterile. A dual infection with Mycobacterium tuberculosis and Mycobacterium avium was diagnosed with multiplex polymerase chain reaction (MPCR. Clarithromycin was added to ATT, and on follow-up at 1 and 3 months, the patient responded well. Molecular methods like MPCR should be exploited for routine diagnosis of high-risk patients.

  16. Association between milk antibody and interferon-gamma responses in cattle from Mycobacterium avium subsp. paratuberculosis infected herds

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Jungersen, Gregers; Nielsen, Søren Saxmose

    2009-01-01

    by use of an IL-12 potentiated IFN-g protocol. Following calving, milk samples were collected and analysed for MAP specific antibodies by ELISA and faecal samples were cultured. The relationship between the variables IFN-g and FC and the outcome of ELISA was assessed using generalised additive models......Paratuberculosis is a chronic infection of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). It is possible to detect infection with paratuberculosis at different stages of disease by means of various diagnostic test strategies. The objective of the present study...... was to evaluate if early cell-mediated immunity could predict the antibody results of milk samples in cattle with different faecal culture (FC) status. A group of 975 cows from 18 Danish MAP infected dairy herds was studied during a 3-year period. Cell-mediated immunity was measured in blood samples from heifers...

  17. Longitudinal Pathogenesis Study of Young Red Deer (Cervus elaphus after Experimental Challenge with Mycobacterium avium subsp. paratuberculosis (MAP

    Directory of Open Access Journals (Sweden)

    Colin Mackintosh

    2012-01-01

    Full Text Available Paratuberculosis progresses more quickly in young red deer than in sheep or cattle. This study describes the clinical, immunological and pathological changes over a 50-week period in fourteen 4-month-old red deer that received heavy oral challenge with Mycobacterium avium subsp. paratuberculosis (MAP. At 4 and 12 weeks post challenge they were anaesthetized and a section of jejunal lymph node was surgically removed for culture, histopathology, and genetic studies. All 14 deer became infected, none were clinically affected, and they had varying degrees of subclinical disease when killed at week 50. Week 4 biopsies showed no paratuberculosis lesions, but MAP was cultured from all animals. At weeks 12 and 50 histopathological lesions ranged from mild to severe with corresponding low-to-high antibody titres, which peaked at 12–24 weeks. IFN-γ responses peaked at 8–15 weeks and were higher in mildly affected animals than in those with severe lesions.

  18. [Hypersensitivity pneumonitis-like disease caused by exposure to Mycobacterium avium complex in bathtub water at home: a case report].

    Science.gov (United States)

    Kanno, Kiyoshi; Akai, Masaya; Kato, Tomohiro; Tada, Toshihiko; Watanabe, Kizuku; Shiozaki, Kouhei; Hase, Mitsuo

    2012-05-01

    We report here a case of hypersensitivity pneumonitis-like disease in an adult, likely due to exposure to Mycobacterium avium complex (MAC) in his bathtub water at home. A 63-year-old man was referred to our hospital with exertional dyspnea. Chest computed tomography showed bilateral, diffuse, centrilobular ground-glass nodules. Bronchoalveolar lavage showed marked lymphocytosis. Transbronchial biopsy showed epithelioid cell granulomas and lymphocyte alveolitis. Cultures of the patient's sputum and bathtub water yielded MAC. Variable-number tandem repeat analysis of the MAC strains in the sputum and bathtub water samples showed that the strains were genetically identical. The clinical condition of the patient improved at home under chemotherapy by avoiding the use of the bathtub.

  19. Characterization of microsatellites in wild and sweet cherry (Prunus avium L.)--markers for individual identification and reproductive processes.

    Science.gov (United States)

    Schueler, Silvio; Tusch, Alexandra; Schuster, Mirko; Ziegenhagen, Birgit

    2003-02-01

    Nuclear microsatellites were characterized in Prunus avium and validated as markers for individual and cultivar identification, as well as for studies of pollen- and seed-mediated gene flow. We used 20 primer pairs from a simple sequence repeat (SSR) library of Prunus persica and identified 7 loci harboring polymorphic microsatellite sequences in P. avium. In a natural population of 75 wild cherry trees, the number of alleles per locus ranged from 4 to 9 and expected heterozygosity from 0.39 to 0.77. The variability of the SSR markers allowed an unambiguous identification of individual trees and potential root suckers. Additionally, we analyzed 13 sweet cherry cultivars and differentiated 12 of them. An exclusion probability of 0.984 was calculated, which indicates that the seven loci are suitable markers for paternity analysis. The woody endocarp was successfully used for resolution of all microsatellite loci and exhibited the same multilocus genotype as the mother tree, as shown in a single seed progeny. Hence, SSR fingerprinting of the purely maternal endocarp was also successful in this Prunus species, allowing the identification of the mother tree of the dispersed seeds. The linkage of microsatellite loci with PCR-amplified alleles of the self-incompatibility locus was tested in two full-sib families of sweet cherry cultivars. From low recombination frequencies, we inferred that two loci are linked with the S locus. The present study provides markers that will significantly facilitate studies of spatial genetic variation and gene flow in wild cherry, as well as breeding programs in sweet cherry.

  20. The role of polar auxin transport through pedicels of Prunus avium L. in relation to fruit development and retention.

    Science.gov (United States)

    Else, Mark A; Stankiewicz-Davies, Anna P; Crisp, Carol M; Atkinson, Christopher J

    2004-09-01

    It was investigated whether premature fruit abscission in Prunus avium L. was triggered by a reduction in polar auxin transport (PAT). The capacity of pedicels to transport tritiated IAA ([3H]-IAA) via the PAT pathway was measured at intervals throughout flower and fruit development. The extent of passive diffusion, assessed by concurrent applications of [14C]-benzoic acid ([14C]-BA), was negligible. Transported radioactivity recovered from agar blocks eluted at the same retention time as authentic [3H]-IAA during HPLC fractionation. The capacity for PAT was already high 7 d before anthesis and increased further following the fertilization of flowers at anthesis. PAT intensity was greatest immediately following fertilization and at the beginning of the cell expansion phase of fruit growth; the transport intensity in fruitlets destined to abscind was negligible. The amount of endogenous IAA moving through the PAT pathway was greatest during the first 3 weeks after fertilization and was again high at the beginning of the fruit expansion stage. IAA export in the phloem increased following fertilization then declined below detectable levels. ABA export in the phloem increased markedly during stone formation and at the onset of fruit expansion. TIBA applied to pedicels of fruit in situ promoted fruitlet abscission in 2000 but not in 2001, despite PAT capacity being reduced by over 98% in the treated pedicels. The application of TIBA to pedicels did not affect fruit expansion. The role of PAT and IAA in relation to the development and retention of Prunus avium fruit is discussed.

  1. Diversità funzionale in cloni di ciliegio da legno (Prunus avium L. di provenienza Appennino toscano

    Directory of Open Access Journals (Sweden)

    Andrea Cutini

    2009-12-01

    Full Text Available Functionality in wild cherry (Prunus avium L. clones of Tuscany Appenines provenances. Results of a research regarding the functionality of already selected wild cherry (Prunus avium L. clones are reported. The main target was to select the genotypes with the best ecological efficiency and less sensible to environmental stress, in order to give concrete indications for arboriculture for wood productions. Starting from 2002, measurements were carried out in the experimental plot of Papiano (Stia, AR, where the following clones with provenance from the Tuscan Apennines were compared: Casina Alpe 1 (A, Casina Alpe 2 (D, Puzzòlo (C, Paradisino (E, Piantata Catenaia (F. Dendrometrical data were collected at the beginning and at the end of each season, in order to evaluate the growth and the individual current increment of the clones. To better characterize the canopies of each clone, measurements of photosynthetic active radiation (PAR and of the leaf area index (LAI where carried out with ceptometers and PCA LAI 2000. In order to evaluate differences between the clones regarding functionality and response to environmental stress, growth and productivity were related to the most important canopy characteristics. Ecological efficiency was calculated for the different clones using the net assimilation rate (NAR. The results show that the clone E has the most developed canopies and the best results in terms of growth. But at the same time it also presents densely branched round canopies and results more sensible to the effects of summer drought. These elements contribute to advise against the use of this clone in future genetic improvement programs and in high quality wood productions. On the contrary, the clones C and A have both good growth characteristics and a better general architecture and are therefore advised for high quality wood productions especially in the same geographic region.

  2. Possible transmission of Mycobacterium avium subspecies paratuberculosis through potable water: lessons from an urban cluster of Crohn's disease

    Directory of Open Access Journals (Sweden)

    Pierce Ellen S

    2009-09-01

    Full Text Available Abstract A "cluster" of patients refers to the geographic proximity of unrelated patients with the same disease and suggests a common environmental cause for that disease. Clusters of patients with Crohn's disease have been linked to the presence of an infectious microorganism in unpasteurized milk and cheese, untreated water supplied by wells or springs, animal manure used as fertilizer for family vegetable gardens, and bodies of water contaminated by agricultural runoff. Mycobacterium avium subspecies paratuberculosis (MAP is the suspected cause of Crohn's disease. MAP causes a disease in dairy cows and other animals that is similar to Crohn's disease, called Johne's ('Yo-knees' disease or paratuberculosis. Dairy cows with Johne's disease secrete MAP into their milk and excrete MAP into their feces. MAP is present in untreated water such as well water, in bodies of water contaminated by agricultural runoff, and in unpasteurized milk and cheese. The "treatment" of "tap" water to make it "drinkable" or "potable" by the processes of sedimentation, filtration and chlorination has little to no effect on MAP. MAP is so resistant to chlorine disinfection that such disinfection actually selects for its growth. Other subspecies of Mycobacterium avium grow in biofilms present on tap water pipes. Despite the documented presence of MAP in tap water and its probable growth on tap water pipes, clusters of Crohn's disease have not previously been described in relationship to tap water pipes supplying patients' homes. This report describes three unrelated individuals who lived on the same block along a street in a midwestern American city and developed Crohn's disease within four years of each other in the 1960's. A common tap water pipe supplied their homes. This is the first reported cluster of Crohn's disease possibly linked to fully treated drinking water, and is consistent with previously reported clusters of Crohn's disease linked to an infectious

  3. A METHOD OF STIMULATING ORIGINAL THINKING IN COLLEGE STUDENTS.

    Science.gov (United States)

    ROBERTSON, MALCOLM H.

    AN ATTEMPT WAS MADE TO SUBSTANTIATE THE HYPOTHESES THAT SUBJECTS RECEIVING SENSORY DEPRIVATION WOULD SHOW MORE IMPROVEMENT IN ORIGINALITY THAN THOSE EXPOSED TO A NORMAL SENSORY ENVIRONMENT, AND THOSE SUBJECTS RECEIVING 4 HOURS OF ISOLATION WOULD SHOW MORE IMPROVEMENT IN ORIGINALITY THAN THOSE RECEIVING 2 HOURS OF ISOLATION. ABOUT 60 VOLUNTEER,…

  4. Cellular immune responses to ESAT-6 discriminate between patients with pulmonary disease due to Mycobacterium avium complex and those with pulmonary disease due to Mycobacterium tuberculosis

    DEFF Research Database (Denmark)

    Lein, A D; von Reyn, C F; Ravn, P;

    1999-01-01

    ESAT-6 (for 6-kDa early secreted antigenic target) is a secreted antigen found almost exclusively in organisms of the Mycobacterium tuberculosis complex. We compared in vitro gamma interferon (IFN-gamma) responses by peripheral blood mononuclear cells to this antigen in patients with pulmonary...... disease due to either Mycobacterium avium complex (MAC) or Mycobacterium tuberculosis with those in healthy, skin test-negative, control subjects. Significant IFN-gamma responses to ESAT-6 were detected in 16 (59%) of 27 M. tuberculosis pulmonary disease patients, 0 (0%) of 8 MAC disease patients, and 0...... (0%) of 8 controls. Significant IFN-gamma responses to M. tuberculosis purified protein derivative were detected in 23 (85%) of 27 M. tuberculosis disease patients, 2 (25%) of 8 MAC disease patients, and 5 (63%) of 8 healthy controls. M. avium sensitin was recognized in 24 (89%) of 27 M. tuberculosis...

  5. Development and validation of a liquid medium (M7H9C) for routine culture of Mycobacterium avium subsp. paratuberculosis to replace modified Bactec 12B medium.

    Science.gov (United States)

    Whittington, Richard J; Whittington, Ann-Michele; Waldron, Anna; Begg, Douglas J; de Silva, Kumi; Purdie, Auriol C; Plain, Karren M

    2013-12-01

    Liquid culture of Mycobacterium avium subsp. paratuberculosis from clinical samples, such as feces, is the most sensitive antemortem test for the diagnosis of Johne's disease in ruminants. In Australia, New Zealand, the United States, and some other countries, the Bactec 460 system with modified Bactec 12B medium (Becton, Dickinson) has been the most commonly used liquid culture system, but it was discontinued in 2012. In this study, a new liquid culture medium, M7H9C, was developed. It consists of a Middlebrook 7H9 medium base with added Casitone, albumin, dextrose, catalase, egg yolk, mycobactin J, and a cocktail of antibiotics. We found that polyoxyethylene stearate (POES) was not essential for the cultivation of M. avium subsp. paratuberculosis in either the Bactec 12B or the M7H9C medium. The limit of detection determined using pure cultures of the C and S strains of M. avium subsp. paratuberculosis was 7 bacilli per 50 μl inoculum in the two media. The new medium was validated using 784 fecal and tissue samples from sheep and cattle, >25% of which contained viable M. avium subsp. paratuberculosis. Discrepant results for the clinical samples between the two media were mostly associated with samples that contained <10 viable bacilli per gram, but these results were relatively uncommon, and the performances of the two media were not significantly different. M7H9C medium was less than half the cost of the Bactec 12B medium and did not require regular examination during incubation, but a confirmatory IS900 PCR test had to be performed on every culture after the predetermined incubation period.

  6. Transcriptional dynamics of the developing sweet cherry (Prunus avium L.) fruit: sequencing, annotation and expression profiling of exocarp-associated genes

    OpenAIRE

    Merianne Alkio; Uwe Jonas; Myriam Declercq; Steven van Nocker; Moritz Knoche

    2014-01-01

    The exocarp, or skin, of fleshy fruit is a specialized tissue that protects the fruit, attracts seed dispersing fruit eaters, and has large economical relevance for fruit quality. Development of the exocarp involves regulated activities of many genes. This research analyzed global gene expression in the exocarp of developing sweet cherry (Prunus avium L., ‘Regina’), a fruit crop species with little public genomic resources. A catalog of transcript models (contigs) representing expressed genes...

  7. Cellular and humoral immune responses in sheep vaccinated with candidate antigens MAP2698c and MAP3567 from Mycobacterium avium subspecies paratuberculosis

    OpenAIRE

    Ratna eGurung; Auriol ePurdie; Richard eWhittington; Douglas eBegg

    2014-01-01

    Control of Johne's disease, caused by Mycobacterium avium subspecies paratuberculosis (MAP) in ruminants using commercially available vaccine reduces production losses, mortality, fecal shedding and histopathological lesions but does not provide complete protection from infection and interferes with serological diagnosis of Johne's disease and bovine tuberculosis. At this time no recombinant antigens have been found to provide superior protection compared to whole killed or live-attenuated MA...

  8. First detection of AmpC β-lactamase bla(CMY-2) on a conjugative IncA/C plasmid in a Vibrio parahaemolyticus isolate of food origin.

    Science.gov (United States)

    Li, Ruichao; Lin, Dachuan; Chen, Kaichao; Wong, Marcus Ho Yin; Chen, Sheng

    2015-07-01

    Vibrio parahaemolyticus is an important causative agent of gastroenteritis, with the consumption of contaminated seafood being the major transmission route. Resistance to penicillin is common among V. parahaemolyticus strains, whereas cephalosporin resistance remains rare. In an attempt to assess the current prevalence and characteristics of antibiotic resistance of this pathogen in common food samples, a total of 54 (17% of the total samples) V. parahaemolyticus strains were isolated from 318 meat and seafood samples purchased from supermarkets and wet markets in Shenzhen, China, in 2013. These isolates exhibited high-level resistance to ampicillin, yet they were mostly susceptible to other antimicrobials, except for two that were resistant to extended-spectrum cephalosporins. The β-lactamase gene blaPER-1 was detectable in one strain, V. parahaemolyticus V43, which was resistant to both third- and fourth-generation cephalosporins. Compared to other blaPER-1-positive V. parahaemolyticus strains reported in our previous studies, strain V43 was found to harbor an ∼200-kb conjugative plasmid carrying genes that were different from the antimicrobial resistance genes reported from the previous studies. The β-lactamase gene blaCMY-2 was detectable for the first time in another V. parahaemolyticus isolate, V4, which was resistant to third-generation cephalosporins. This blaCMY-2 gene was shown to be located in an ∼150-kb IncA/C-type conjugative plasmid with a genetic structure consisting of traB-traV-traA-ISEcp1-blaCMY-2-blc-sugE-encR-orf1-orf2-orf3-orf4-dsbC-traC, which is identical to that of other IncA/C conjugative plasmids in Enterobacteriaceae, albeit with a different size. These findings indicate that the transmission of extended-spectrum-β-lactamase (ESBL) and AmpC β-lactamase genes via conjugative plasmids can mediate the development of extended-spectrum cephalosporin resistance in V. parahaemolyticus, thereby posing a potential threat to public health.

  9. 1株鸽源鸡杆菌复合群3的分离和鉴定%Isolation and identification of one pigeon-origin Gallibacterium genomo sp3

    Institute of Scientific and Technical Information of China (English)

    杨晓林; 陈英; 余松城; 邹年莉; 吴瑞婷; 夏静; 牛婷; 王富妍; 黄勇

    2014-01-01

    报道我国鸽场中鸡杆菌的感染情况.四川某种鸽场的种鸽出现以消瘦、呼吸困难、气管和肺出血、腹膜炎、输卵管炎等为主要特征的疾病.选取代表性病例,进行细菌分离和病毒检测,并对分离菌进行形态学观察、生化鉴定、动物试验、药敏试验和16S rRNA 基因序列分析.结果显示,部分病鸽只检测到新城疫病毒;从肝、肺中分离到1株革兰阴性短杆菌,命名为 SC01,该菌株为致病菌,小鼠半数致死量(LD50)为1.26×109 CFU/mL,能发酵多数糖和醇类,对头孢类药物较敏感.将 SC01与 GenBank 中同源性较近的15株细菌的16S rRNA 基因序列进行同源性分析,发现 SC01株与4株鸡杆菌复合群3(Gallibacterium genomo sp.3)的同源性为98.5%~99.5%,其中与EU423996的同源性最高,为99.5%,证实该菌株为鸡杆菌复合群3.试验结果为鸡杆菌复合群3感染的流行病学调查、诊断与治疗提供了新的参考.%Summary Gallibacterium infection was a new infectious disease observed in some areas of China,which was mainly harmful for laying hen,resulting in salpingitis,oophoritis,peritonitis,septicemia,hepatitis,enteritis, respiratory tract diseases,and so on.So far,most of the G.anatis were isolated from poultry,including chickens, ducks,pigeons,geese,parrots,quail and guinea fowl,etc.Isolation of Gallibacterium from pigeons was not reported in China. A contagious disease occurred in one breeding farm of pigeon in Sichuan Province in January of 2013,with the characteristic signs of depressed wasting,dyspnea,diarrhea,and so on.The morbidity was about 43%,and the mortality was about 18%.The disease was mainly found in female pigeons,and female pigeons of different ages could be infected,especially pigeons of 1 to 1.5 years old.Mild bleeding and mucus could be observed in diseased pigeons,and other gross lesions could also be observed including hemorrhage of lung,peritonitis,salpingitis,mild bleeding of duodenum,and so on

  10. Rhizobium nepotum sp. nov. isolated from tumors on different plant species.

    Science.gov (United States)

    Puławska, Joanna; Willems, Anne; De Meyer, Sofie E; Süle, Sandor

    2012-06-01

    Five Gram-negative, rod-shaped, non-spore-forming bacteria were isolated from galls on different plant species in Hungary: strain 39/7(T) from Prunus cerasifera Myrobalan, strain 0 from grapevine var. Ezerjó, strain 7/1 from raspberry var. Findus and in Poland, strain C3.4.1 from Colt rootstock (Prunus avium × Prunus pseudocerasus) and strain CP17.2.2 from Prunus avium. Only one of these isolates, strain 0, is able to cause crown gall on different plant species. On the basis of 16S rRNA gene sequence similarity, the strains cluster together and belong to the genus Rhizobium and their closest relative is Rhizobium radiobacter (99.1%). Phylogenetic analysis of the novel strains using housekeeping genes atpD, glnA, gyrB, recA and rpoB revealed their distinct position separate from other known Rhizobium species and confirmed their relation to Rhizobium radiobacter. The major cellular fatty acids are 18:1 w7c, 16:0, 16:0 3OH, summed feature 2 (comprising 12:0 aldehyde, 16:1 iso I and/or 14:0 3OH) and summed feature 3 (comprising 16:1 w7c and/or 15 iso 2OH). DNA-DNA hybridization of strain 39/7(T) with the type strain of R. radiobacter LMG 140(T) revealed 45% DNA-DNA hybridization. Phenotypic and physiological properties differentiate the novel isolates from other closely related species. On the basis of the results obtained, the five isolates are considered to represent a novel species of the genus Rhizobium, for which the name Rhizobium nepotum sp. nov. (type strain 39/7(T)=LMG 26435(T)=CFBP 7436(T)) is proposed.

  11. A multilaboratory evaluation of a commercial enzyme-linked immunosorbent assay test for the detection of antibodies against Mycobacterium avium subsp. paratuberculosis in cattle.

    Science.gov (United States)

    Dargatz, David A; Byrum, Beverly A; Collins, Michael T; Goyal, Sagar M; Hietala, Sharon K; Jacobson, Richard H; Kopral, Christine A; Martin, Barbara M; McCluskey, Brian J; Tewari, Deepanker

    2004-11-01

    Five laboratories participated in a study to evaluate sources of variation in results from an enzyme-linked immunosorbent assay (ELISA) for antibodies against Mycobacterium avium subsp. paratuberculosis. Each laboratory repeatedly tested duplicates of a negative, positive (P), and high-positive (HP) serum sample, which were supplied by the United States Department of Agriculture: Animal and Plant Health Inspection Service: Veterinary Services, National Veterinary Services Laboratories, Ames, IA, on all 96-well microtiter plates when routinely testing other samples for M. avium subsp. paratuberculosis antibodies. These 3 sera were aliquoted and sent to the 5 participating laboratories. This study focused on variation in test results because of assay reagents and laboratory techniques and did not account for biologic variability associated with the time course of infection in cattle. Overall, results from 868 microtiter plates were used in the study. For each sample a sample-to-positive (S/P) ratio was calculated according to the manufacturer's directions. The S/ P ratio for the P sample ranged from 0.06 to 1.039 (mean = 0.466 and 0.484 for wells 1 and 2, respectively) and those for the HP sample ranged from 2.446 to 8.727 (mean = 4.027 and 3.980 for wells 1 and 2, respectively). The majority of the variation in S/P ratio for the P sample was attributed to kit lot (37.5%), followed by random (unexplained) error (27.0%), laboratory (18.3%), and kit lot by laboratory (11.9%). By eliminating plates in which the separation between negative and positive control ODs was less than 0.4, the proportion of variation attributed to laboratory was reduced markedly. These results confirm that there is variability in M. avium subsp. paratuberculosis ELISA results and that several sources contribute to the observed variability. The study gives a relative estimate of the contribution of various sources to the overall variability observed in the M. avium subsp. paratuberculosis ELISA

  12. Market study: Biological isolation garment

    Science.gov (United States)

    1975-01-01

    The biological isolation garment was originally designed for Apollo astronauts to wear upon their return to earth from the moon to avoid the possibility of their contaminating the environment. The concept has been adapted for medical use to protect certain patients from environmental contamination and the risk of infection. The nature and size of the anticipated market are examined with certain findings and conclusions relative to clinical acceptability and potential commercial viability of the biological isolation garment.

  13. Integrated microRNA-mRNA-analysis of human monocyte derived macrophages upon Mycobacterium avium subsp. hominissuis infection.

    Directory of Open Access Journals (Sweden)

    Jutta Sharbati

    Full Text Available BACKGROUND: Many efforts have been made to understand basal mechanisms of mycobacterial infections. Macrophages are the first line of host immune defence to encounter and eradicate mycobacteria. Pathogenic species have evolved different mechanisms to evade host response, e.g. by influencing macrophage apoptotic pathways. However, the underlying molecular regulation is not fully understood. A new layer of eukaryotic regulation of gene expression is constituted by microRNAs. Therefore, we present a comprehensive study for identification of these key regulators and their targets in the context of host macrophage response to mycobacterial infections. METHODOLOGY/PRINCIPAL FINDINGS: We performed microRNA as well as mRNA expression analysis of human monocyte derived macrophages infected with several Mycobacterium avium hominissuis strains by means of microarrays as well as quantitative reverse transcription PCR (qRT-PCR. The data revealed the ability of all strains to inhibit apoptosis by transcriptional regulation of BCL2 family members. Accordingly, at 48 h after infection macrophages infected with all M. avium strains showed significantly decreased caspase 3 and 7 activities compared to the controls. Expression of let-7e, miR-29a and miR-886-5p were increased in response to mycobacterial infection at 48 h. The integrated analysis of microRNA and mRNA expression as well as target prediction pointed out regulative networks identifying caspase 3 and 7 as potential targets of let-7e and miR-29a, respectively. Consecutive reporter assays verified the regulation of caspase 3 and 7 by these microRNAs. CONCLUSIONS/SIGNIFICANCE: We show for the first time that mycobacterial infection of human macrophages causes a specific microRNA response. We furthermore outlined a regulatory network of potential interactions between microRNAs and mRNAs. This study provides a theoretical concept for unveiling how distinct mycobacteria could manipulate host cell response

  14. Improved Culture Medium (TiKa) for Mycobacterium avium Subspecies Paratuberculosis (MAP) Matches qPCR Sensitivity and Reveals Significant Proportions of Non-viable MAP in Lymphoid Tissue of Vaccinated MAP Challenged Animals.

    Science.gov (United States)

    Bull, Tim J; Munshi, Tulika; Mikkelsen, Heidi; Hartmann, Sofie B; Sørensen, Maria R; Garcia, Joanna S; Lopez-Perez, Paula M; Hofmann, Sven; Hilpert, Kai; Jungersen, Gregers

    2016-01-01

    The quantitative detection of viable pathogen load is an important tool in determining the degree of infection in animals and contamination of foodstuffs. Current conventional culture methods are limited in their ability to determine these levels in Mycobacterium avium subspecies paratuberculosis (MAP) due to slow growth, clumping and low recoverability issues. The principle goal of this study was to evaluate a novel culturing process (TiKa) with unique ability to stimulate MAP growth from low sample loads and dilutions. We demonstrate it was able to stimulate a mean 29-fold increase in recoverability and an improved sensitivity of up to three logs when compared with conventional culture. Using TiKa culture, MAP clumping was minimal and produced visible colonies in half the time required by standard culture methods. Parallel quantitative evaluation of the TiKa culture approach and qPCR on MAP loads in tissue and gut mucosal samples from a MAP vaccine-challenge study, showe