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Sample records for avium subspecies paratuberculosis

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

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    AdelMTalaat

    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.

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

    DEFF Research Database (Denmark)

    Afzal, Mamuna; Abidi, Soad; Mikkelsen, Heidi; Tang, Sheila Tuyet; Lund, Ole; Nielsen, Morten; Lundegaard, Claus; Jungersen, Gregers; Ussery, David

    We have sequenced a Danish isolate of Mycobacterium avium subspecies paratuberculosis, strain Ejlskov2007. The strain was isolated from faecal material of a 48 month old second parity Danish Holstein cow, with clinical symptoms of chronic diarrhoea and emaciation. The cultures were grown on......, 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 analyses of the sequenced Mycobacterium genomes reveal a surprisingly open and diverse set of genes for this bacterial genera....

  3. Optical mapping of the Mycobacterium avium subspecies paratuberculosis genome

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    Schwartz David C

    2009-01-01

    Full Text Available Abstract Background Infection of cattle with Mycobacterium avium subspecies paratuberculosis (M. ap causes severe economic losses to the dairy industry in the USA and worldwide. In an effort to better examine diversity among M. ap strains, we used optical mapping to profile genomic variations between strains of M. ap K-10 (sequenced strain and M. ap ATCC 19698 (type strain. Results The assembled physical restriction map of M. ap ATCC 19698 showed a genome size of 4,839 kb compared to the sequenced K-10 genome of 4,830 kb. Interestingly, alignment of the optical map of the M. ap ATCC 19698 genome to the complete M. ap K-10 genome sequence revealed a 648-kb inversion around the origin of replication. However, Southern blotting, PCR amplification and sequencing analyses of the inverted region revealed that the genome of M. ap K-10 differs from the published sequence in the region starting from 4,197,080 bp to 11,150 bp, spanning the origin of replication. Additionally, two new copies of the coding sequences > 99.8% were identified, identical to the MAP0849c and MAP0850c genes located immediately downstream of the MAP3758c gene. Conclusion The optical map of M. ap ATCC 19698 clearly indicated the miss-assembly of the sequenced genome of M. ap K-10. Moreover, it identified 2 new genes in M. ap K-10 genome. This analysis strongly advocates for the utility of physical mapping protocols to complement genome sequencing projects.

  4. 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 ...

  5. Characteristics of an Extensive Mycobacterium avium subspecies paratuberculosis Recombinant Protein Set

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    In the first step of a comprehensive large-scale antigen discovery project, 651 Mycobacterium avium subspecies paratuberculosis proteins were produced in Escherichia coli. All of these were purified by affinity chromatography, dialyzed in phosphate buffered saline, and analyzed on SDS-PAGE gels. C...

  6. Survey of Mycobacterium avium subspecies paratuberculosis serological status in beef herds on community pastures in Saskatchewan

    OpenAIRE

    Waldner, Cheryl L.; Cunningham, Gail L.; Janzen, Eugene D.; Campbell, John R.

    2002-01-01

    Johne's disease is a well recognized problem in dairy herds. Relatively little information is available on either the prevalence or the control of Johne's disease in commercial cow-calf operations. In the fall of 1999, blood samples were collected during pregnancy testing from cows on community pastures in Saskatchewan. Sera from these cows were analyzed using a commercial ELISA for antibodies to Mycoplasma avium subspecies paratuberculosis. All cows from each herd examined at the community p...

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

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    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. PMID:25128370

  8. Risk factors for the introduction and within-herd transmission of Mycobacterium avium subspecies paratuberculosis (MAP) infection on 59 Irish dairy herds

    OpenAIRE

    Cashman, W.; Buckley, J.; Quigley, T.; Fanning, S.; S. More; Egan, J.; Berry, D.; GRANT, I.; O'Farrell, K

    2008-01-01

    Since 1994, Irish cattle have been exposed to greater risks of acquiring Mycobacterium avium subspecies paratuberculosis (MAP) infection as a consequence of the importation of over 70,000 animals from continental Europe. In recent years, there has been an increase in the number of reported clinical cases of paratuberculosis in Ireland. This study examines the prevalence of factors that promote the introduction and within-herd transmission of Mycobacterium avium subspecies paratuberculosis (MA...

  9. Early Antibody Response Against Mycobacterium avium subspecies paratuberculosis Antigens in Subclinical Cattle

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    Abstract Background Our laboratories have previously reported on the experimental infection of cattle with Mycobacterium avium subsp paratuberculosis (M. paratuberculosis) using an intratonsillar infection model. In addition, we have recently developed a partial protein array representing 92 M. par...

  10. Genome-Wide Association Analysis and Genomic Prediction of Mycobacterium avium Subspecies paratuberculosis Infection in US Jersey Cattle

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    Zare, Yalda; Shook, George E.; Collins, Michael T.; Kirkpatrick, Brian W.

    2014-01-01

    Paratuberculosis (Johne’s disease), an enteric disorder in ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP), causes economic losses in excess of $200 million annually to the US dairy industry. To identify genomic regions underlying susceptibility to MAP infection in Jersey cattle, a case-control genome-wide association study (GWAS) was performed. Blood and fecal samples were collected from ∼5,000 mature cows in 30 commercial Jersey herds from across the US. Discovery ...

  11. Characterization of the inflammatory phenotype of Mycobacterium avium subspecies paratuberculosis using a novel cell culture passage model

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    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. Longitudinal data collection of Mycobacterium avium subspecies Paratuberculosis infections in dairy herds. Collection and use of observational data

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    Longitudinal infection data on Mycobacterium avium subspecies paratuberculosis (MAP) was collected on three dairy farms in Northeastern United States during approximately 10 years. Precise data on animal characteristics and animal location within farm were collected on these farms. Cows were followe...

  13. 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,...

  14. CD4 T Cells From Intestinal Biopsies of Crohn's Disease Patients React to Mycobacterium avium subspecies paratuberculosis

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    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...

  15. Genotype profiles of Mycobacterium avium subspecies paratuberculosis recovered from suspected and Crohn's disease patients in India.

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    Singh, A V; Singh, S V; Sohal, J S; Singh, P K

    2010-06-01

    Present study aimed to genotype Mycobacterium avium subspecies paratuberculosis (MAP) recovered from suspected and Crohn' s disease patients. A total of 32 MAP and DNA (directly from clinical samples) recovered from human origin were genotyped using IS 1311 PCR-REA. Isolates were cultured from stool, biopsies and blood clots of Crohn's disease patients, and stool samples of suspected (animal attendants, lab workers etc). Of the 32 MAP isolates belonging to 28 human beings, majority (84.3%) were genotyped as 'Bison type', while 21.7% were of 'cattle' and none was 'sheep' genotype. Study first time reports distribution of 'Cattle' and 'Bison type' 'genotypes in suspected and Crohn's patients on pilot scale in India. 'Bison type' genotype was predominant in the surveyed human population. PMID:22471168

  16. Genome sequencing of ovine isolates of Mycobacterium avium subspecies paratuberculosis offers insights into host association

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    Bannantine John P

    2012-03-01

    Full Text Available Abstract Background The genome of Mycobacterium avium subspecies paratuberculosis (MAP is remarkably homogeneous among the genomes of bovine, human and wildlife isolates. However, previous work in our laboratories with the bovine K-10 strain has revealed substantial differences compared to sheep isolates. To systematically characterize all genomic differences that may be associated with the specific hosts, we sequenced the genomes of three U.S. sheep isolates and also obtained an optical map. Results Our analysis of one of the isolates, MAP S397, revealed a genome 4.8 Mb in size with 4,700 open reading frames (ORFs. Comparative analysis of the MAP S397 isolate showed it acquired approximately 10 large sequence regions that are shared with the human M. avium subsp. hominissuis strain 104 and lost 2 large regions that are present in the bovine strain. In addition, optical mapping defined the presence of 7 large inversions between the bovine and ovine genomes (~ 2.36 Mb. Whole-genome sequencing of 2 additional sheep strains of MAP (JTC1074 and JTC7565 further confirmed genomic homogeneity of the sheep isolates despite the presence of polymorphisms on the nucleotide level. Conclusions Comparative sequence analysis employed here provided a better understanding of the host association, evolution of members of the M. avium complex and could help in deciphering the phenotypic differences observed among sheep and cattle strains of MAP. A similar approach based on whole-genome sequencing combined with optical mapping could be employed to examine closely related pathogens. We propose an evolutionary scenario for M. avium complex strains based on these genome sequences.

  17. Seroprevalence and risk factors of Mycobacterium avium subspecies paratuberculosis infection in domestic sika deer in China.

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    Meng, Qing-Feng; Li, Ying; Yang, Fan; Yao, Gui-Zhi; Qian, Ai-Dong; Wang, Wei-Li; Cong, Wei

    2015-06-01

    Paratuberculosis or Johne's disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic infectious granulomatous enteritis of ruminants and other animals, which has a worldwide occurrence, but little is known of MAP infection in domestic sika deer in Jilin Province, China. The objective of the present investigation was to examine seroprevalence and risk factors of MAP infection in Jilin Province. Serum samples collected from 1400 sika deer from 16 sika deer herds were collected in the 4 districts of the province between May 2013 and August 2014 and were tested independently for the presence of antibodies against MAP. A total of 247 (17.64 %) sika deer tested positive for MAP antibodies using a commercially available enzyme immunoassay kit. The management level of farm and collecting region of sika deer was the main risk factor associated with MAP infection. The present study revealed the seroprevalence of MAP infection in sika deer in Jilin Province, China, which provided the baseline data for taking comprehensive countermeasures and measures in effectively preventing and controlling MAP infection in sika deer. PMID:25904509

  18. Mycobacterium avium Subspecies paratuberculosis Infection Modifies Gut Microbiota under Different Dietary Conditions in a Rabbit Model

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    Arrazuria, Rakel; Elguezabal, Natalia; Juste, Ramon A.; Derakhshani, Hooman; Khafipour, Ehsan

    2016-01-01

    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 to be 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 infection

  19. Mycobacterium avium subspecies paratuberculosis infection modifies gut microbiota under different dietary conditions in a rabbit model

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    Rakel Arrazuria

    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

  20. Impact of the shedding level on transmission of persistent infections in Mycobacterium avium subspecies paratuberculosis (MAP).

    Science.gov (United States)

    Slater, Noa; Mitchell, Rebecca Mans; Whitlock, Robert H; Fyock, Terry; Pradhan, Abani Kumar; Knupfer, Elena; Schukken, Ynte Hein; Louzoun, Yoram

    2016-01-01

    Super-shedders are infectious individuals that contribute a disproportionate amount of infectious pathogen load to the environment. A super-shedder host may produce up to 10,000 times more pathogens than other infectious hosts. Super-shedders have been reported for multiple human and animal diseases. If their contribution to infection dynamics was linear to the pathogen load, they would dominate infection dynamics. We here focus on quantifying the effect of super-shedders on the spread of infection in natural environments to test if such an effect actually occurs in Mycobacterium avium subspecies paratuberculosis (MAP). We study a case where the infection dynamics and the bacterial load shed by each host at every point in time are known. Using a maximum likelihood approach, we estimate the parameters of a model with multiple transmission routes, including direct contact, indirect contact and a background infection risk. We use longitudinal data from persistent infections (MAP), where infectious individuals have a wide distribution of infectious loads, ranging upward of three orders of magnitude. We show based on these parameters that the effect of super-shedders for MAP is limited and that the effect of the individual bacterial load is limited and the relationship between bacterial load and the infectiousness is highly concave. A 1000-fold increase in the bacterial contribution is equivalent to up to a 2-3 fold increase in infectiousness. PMID:26925966

  1. Mycobacterium avium subspecies paratuberculosis in the etiology of Crohn's disease, cause or epiphenomenon?

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    Liverani, Elisa; Scaioli, Eleonora; Cardamone, Carla; Dal Monte, Paola; Belluzzi, Andrea

    2014-09-28

    The origin of inflammatory bowel disease is unknown. Attempts have been made to isolate a microorganism that could explain the onset of inflammation, but no pathological agent has ever been identified. Johne's disease is a granulomatous chronic enteritis of cattle and sheep caused by Mycobacterium avium subspecies paratuberculosis (MAP) and shows some analogies with Crohn's disease (CD). Several studies have tried to clarify if MAP has a role in the etiology of CD. The present article provides an overview of the evidence in favor and against the "MAP-hypothesis", analyzing the methods commonly adopted to detect MAP and the role of antimycobacterial therapy in patients with inflammatory bowel disease. Studies were identified through the electronic database, MEDLINE, and were selected based on their relevance to the objective of the review. The presence of MAP was investigated using multiple diagnostic methods for MAP detection and in different tissue samples from patients affected by CD or ulcerative colitis and in healthy controls. On the basis of their studies, several authors support a close relationship between MAP and CD. Although increasing evidence of MAP detection in CD patients is unquestionable, a clear etiological link still needs to be proven. PMID:25278700

  2. Mycobacterium avium subspecies paratuberculosis causes Crohn's disease in some inflammatory bowel disease patients.

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    Naser, Saleh A; Sagramsingh, Sudesh R; Naser, Abed S; Thanigachalam, Saisathya

    2014-06-21

    Crohn's disease (CD) is a chronic inflammatory condition that plagues millions all over the world. This debilitating bowel disease can start in early childhood and continue into late adulthood. Signs and symptoms are usually many and multiple tests are often required for the diagnosis and confirmation of this disease. However, little is still understood about the cause(s) of CD. As a result, several theories have been proposed over the years. One theory in particular is that Mycobacterium avium subspecies paratuberculosis (MAP) is intimately linked to the etiology of CD. This fastidious bacterium also known to cause Johne's disease in cattle has infected the intestines of animals for years. It is believed that due to the thick, waxy cell wall of MAP it is able to survive the process of pasteurization as well as chemical processes seen in irrigation purification systems. Subsequently meat, dairy products and water serve as key vehicles in the transmission of MAP infection to humans (from farm to fork) who have a genetic predisposition, thus leading to the development of CD. The challenges faced in culturing this bacterium from CD are many. Examples include its extreme slow growth, lack of cell wall, low abundance, and its mycobactin dependency. In this review article, data from 60 studies showing the detection and isolation of MAP by PCR and culture techniques have been reviewed. Although this review may not be 100% comprehensive of all studies, clearly the majority of the studies overwhelmingly and definitively support the role of MAP in at least 30%-50% of CD patients. It is very possible that lack of detection of MAP from some CD patients may be due to the absence of MAP role in these patients. The latter statement is conditional on utilization of methodology appropriate for detection of human MAP strains. Ultimately, stratification of CD and inflammatory bowel disease patients for the presence or absence of MAP is necessary for appropriate and effective

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

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    Wynne, James W; Bull, Tim J; Seemann, Torsten; Bulach, Dieter M; Wagner, Josef; Kirkwood, Carl D; Michalski, Wojtek P

    2011-01-01

    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. PMID:21799786

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

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    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.

  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 and characterization of Mycobacterium avium subspecies paratuberculosis in Korean black goat (Capra hircus aegagrus

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    JM Kim

    2015-01-01

    Full Text Available Paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis (MAP, is a chronic incurable infection of intestinal tract of animals. The study took place in 2012, on 491 Korean black goats, older than 2 years, from goat herds located in Southern region, Korea. The animals were sampled and tested for paratuberculosis. Enzyme-linked immunosorbent assay and microbiological culture were used to investigate MAP infection. Among the 491 goats from 100 herds screened, 4 (0.8% and 3 (0.6% goats were defined positive for MAP by ELISA and faecal culture, respectively. Isolated colonies were confirmed as MAP by IS900-PCR, and genotyped by IS1311 PCR-REA assays. The results revealed MAP infection at a low prevalence rate with "cattle type" and "bison type" genotypes.

  7. Genomic Characterization of the Vaccinal Strain of Mycobacterium Avium Subspecies Paratuberculosis (MAP 316F by MIRU-VNTR

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    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

  8. Seroprevalence of antibodies against bovine leukemia virus, bovine viral diarrhea virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum in dairy cattle in Saskatchewan

    OpenAIRE

    Vanleeuwen, John A.; Forsythe, LeeAnn; Tiwari, Ashwani; Chartier, Renee

    2005-01-01

    Blood was drawn from 1530 dairy cows in 51 herds. For antibodies against bovine leukemia virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum, 37.4%, 2.7%, and 5.6% of cows were test positive, respectively, while 29.2% of herds had unvaccinated animals with ≥ 1:64 for bovine viral diarrhea virus.

  9. Characterization of gene expression profiles to chronic infection with Mycobacterium avium subspecies paratuberculosis

    DEFF Research Database (Denmark)

    Melvang, Heidi Mikkelsen; Grønbæk, Betina Chemnitz; Brogaard, Louise;

    Mycobacterium avium subsp. paratuberculosis (Map) causes paratuberculosis, a chronic enteritis of ruminants. The aim of the study was to use high-throughput reverse transcriptase (RT) qPCR to describe intestinal gene expression patterns in response to different levels of Map infection with a large...... expected, leads to increased expression of local IFN-γ. Expression of IL-10 also increased as a result of Map infection, and this increase was more correlated to the amount of Map than IFN-γ, indicating a shift towards a regulatory environment as infection progress. Th17-mediated immune responses were...... suppressed at this stage. Gene expression of all other genes could not be interpreted in relation to infection status. High throughput RT qPCR can be used for exploring gene expression patterns in response to Map infection but larger study groups are needed to fully understand which are key mechanisms and...

  10. Risk factors for the introduction and within-herd transmission of Mycobacterium avium subspecies paratuberculosis (MAP) infection on 59 Irish dairy herds

    OpenAIRE

    Cashman W; Buckley J.; Quigley T; Fanning S; More S; Egan J; Berry D.; Grant I; O'Farrell K

    2008-01-01

    Abstract Since 1994, Irish cattle have been exposed to greater risks of acquiring Mycobacterium avium subspecies paratuberculosis (MAP) infection as a consequence of the importation of over 70,000 animals from continental Europe. In recent years, there has been an increase in the number of reported clinical cases of paratuberculosis in Ireland. This study examines the prevalence of factors that promote the introduction and within-herd transmission of Mycobacterium avium subspecies paratubercu...

  11. Seroprevalences of antibodies against bovine leukemia virus, bovine viral diarrhea virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum in beef and dairy cattle in Manitoba

    OpenAIRE

    Vanleeuwen, John A.; Tiwari, Ashwani; Plaizier, Jan C; Whiting, Terry L.

    2006-01-01

    Of 1204 dairy cows and 1425 beef cows sampled, 60.8% and 10.3% were seropositive for Bovine leukemia virus, 4.5% and 1.7% were seropositive for Mycobacterium avium subspecies paratuberculosis, and 8.3% and 9.1% were seropositive for Neospora caninum, respectively, while 28.1% of dairy herds had unvaccinated animals with titres ≥ 1:64 for Bovine viral diarrhea virus.

  12. Evaluation of “Indigenous Absorbed ELISA Kit” for the Estimation of Seroprevalence of Mycobacterium avium Subspecies paratuberculosis Antibodies in Human Beings in North India

    OpenAIRE

    Singh, A. V.; Singh, S. V.; D K Verma; Yadav, R.; P K Singh; Sohal, J. S.

    2011-01-01

    In present pilot study aimed to estimate, presence of Mycobacterium avium subspecies paratuberculosis (MAP) antibodies in the human serum samples originating from North India using “Indigenous absorbed ELISA kit” (ELISA kit). The phase I, “ELISA kit” was optimized using protoplasmic antigen from native isolate of MAP “Indian Bison type” recovered from the biopsies of Crohn's disease patients. The phase II, sensitivity and specificity of the kit were estimated as 40.0 and 83.3%, respectively, ...

  13. 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

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    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

  14. Mycobacterium avium subspecies Paratuberculosis and crohn′s regional ileitis: How strong is association?

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    Sarman Singh

    2011-01-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (MAP is a well-established etiological agent of Johne′s disease in animals. In humans, similar clinical condition, first described by Crohn as regional ileitis in 1932, now known as Crohn′s diseases (CD, has also been associated with this mycobacterial species. However, there are two schools of thoughts, one favoring MAP as its etiological agent while the second considers it as an immune-inflammatory condition triggered by an external factor. Onset of CD requires a series of events including predisposition of certain inherited genetic traits, associated environmental stimuli, and immune-inflammatory response. A combination of these factors probably leads to this disease. Recently, some human genes have also been identified which regulate ability to respond appropriately to the external factors. Added to these factors are concerns about the selection of clinical specimens and poor adherence to laboratory quality controls. The literature is full of contradictory findings, but there a lack of uniformity in the materials and methods used by many of these researchers. In this review, we provide our perspective under above circumstances and give our point of view which may open a platform for debate regarding the MAP as the etiological agent of human CD.

  15. Mycobacterium avium subspecies Paratuberculosis and Crohn's regional ileitis: how strong is association?

    Science.gov (United States)

    Singh, Sarman; Gopinath, Krishnamoorthy

    2011-07-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is a well-established etiological agent of Johne's disease in animals. In humans, similar clinical condition, first described by Crohn as regional ileitis in 1932, now known as Crohn's diseases (CD), has also been associated with this mycobacterial species. However, there are two schools of thoughts, one favoring MAP as its etiological agent while the second considers it as an immune-inflammatory condition triggered by an external factor. Onset of CD requires a series of events including predisposition of certain inherited genetic traits, associated environmental stimuli, and immune-inflammatory response. A combination of these factors probably leads to this disease. Recently, some human genes have also been identified which regulate ability to respond appropriately to the external factors. Added to these factors are concerns about the selection of clinical specimens and poor adherence to laboratory quality controls. The literature is full of contradictory findings, but there a lack of uniformity in the materials and methods used by many of these researchers. In this review, we provide our perspective under above circumstances and give our point of view which may open a platform for debate regarding the MAP as the etiological agent of human CD. PMID:22219557

  16. 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.

  17. Effects of seasonal climatic conditions on the diagnosis of Mycobacterium avium subspecies paratuberculosis in dairy cattle.

    Science.gov (United States)

    Strickland, S J; Scott, H M; Libal, M C; Roussel, A J; Jordan, E R

    2005-07-01

    Validity of Johne's disease programs and control protocols that rely on established cut points [e.g., specified sample-to-positive (S/P) ratios] for ELISA serological tests depends on interpreted results that are not susceptible to variable test accuracy. It was hypothesized that seasonal variability exists in serological response to Mycobacterium avium subsp. paratuberculosis (MAP) infection. Further, a reciprocal response may occur, resulting in greater risk of fecal shedding in subclinically infected animals. A testing regimen was invoked that included multiple testing of individual adult cows during the 4 seasons. Serum was collected on a cyclic, monthly basis from 3 randomly selected cohorts of dairy cows, and fecal samples were collected from the 20% of cows with the greatest ELISA test S/P ratios. Staggered, quarterly sampling was continued for 1 yr, and at the conclusion, serum was analyzed en masse. The ELISA outcome values (i.e., S/P ratio) were treated both as categorical and continuous variables. The potential lagged effects of temperature-related seasonality on S/P ratio, as well as the potential for a change in test result caused by temperature were assessed. Results for fecal culture were analyzed on a categorical scale and compared with the ELISA results to explore the possibility of reciprocal fecal shedding. No significant seasonal effects on either S/P ratios or the proportion of cows seropositive to MAP were observed. Furthermore, no evidence was found linking temperature-related seasonality to a reciprocal increase in the risk of fecal culture positivity for MAP. PMID:15956306

  18. NlpC/P60 domain-containing proteins of Mycobacterium avium subspecies paratuberculosis that differentially bind and hydrolyze peptidoglycan.

    Science.gov (United States)

    Bannantine, John P; Lingle, Cari K; Adam, Philip R; Ramyar, Kasra X; McWhorter, William J; Stabel, Judith R; Picking, William D; Geisbrecht, Brian V

    2016-04-01

    A subset of proteins containing NlpC/P60 domains are bacterial peptidoglycan hydrolases that cleave noncanonical peptide linkages and contribute to cell wall remodeling as well as cell separation during late stages of division. Some of these proteins have been shown to cleave peptidoglycan in Mycobacterium tuberculosis and play a role in Mycobacterium marinum virulence of zebra fish; however, there are still significant knowledge gaps concerning the molecular function of these proteins in Mycobacterium avium subspecies paratuberculosis (MAP). The MAP genome sequence encodes five NlpC/P60 domain-containing proteins. We describe atomic resolution crystal structures of two such MAP proteins, MAP_1272c and MAP_1204. These crystal structures, combined with functional assays to measure peptidoglycan cleavage activity, led to the observation that MAP_1272c does not have a functional catalytic core for peptidoglycan hydrolysis. Furthermore, the structure and sequence of MAP_1272c demonstrate that the catalytic residues normally required for hydrolysis are absent, and the protein does not bind peptidoglycan as efficiently as MAP_1204. While the NlpC/P60 catalytic triad is present in MAP_1204, changing the catalytic cysteine-155 residue to a serine significantly diminished catalytic activity, but did not affect binding to peptidoglycan. Collectively, these findings suggest a broader functional repertoire for NlpC/P60 domain-containing proteins than simply hydrolases. PMID:26799947

  19. Presence of intestinal Mycobacterium avium subspecies paratuberculosis (MAP DNA is not associated with altered MMP expression in ulcerative colitis

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    Halwe Jörg M

    2011-04-01

    Full Text Available Abstract Background Mycobacterium avium subspecies paratuberculosis (MAP is suspected to be a causative agent in human Crohn's disease (CD. Recent evidence suggests that pathogenic mycobacteria and MAP can induce the expression of Matrix Metalloproteinases (MMP, which are the main proteases in the pathogenesis of mucosal ulcerations in inflammatory bowel disease (IBD. Within this study we assessed the prevalence of intestinal MAP specific DNA in patients with Crohn's disease, ulcerative colitis (UC, and healthy controls. We further analysed regulation patterns of MMPs in mucosal tissues of UC patients with and without intestinal MAP DNA detection. Methods Colonic biopsy samples were obtained from 63 Norwegian and German IBD patients and 21 healthy controls. RNA was quantified by quantitative real-time polymerase chain reaction (PCR to study MMP gene expression in both pathological and healthy mucosal specimens. The presence of MAP DNA in colonic mucosa was examined using MAP specific PCR. Results MAP DNA was detected in 20% of UC patients and 33% of healthy controls but only in 7% of patients with CD. UC patients treated with corticosteroids exhibited a significantly increased frequency of intestinal MAP DNA compared to those not receiving corticosteroids. Expression of MMP-1, -2, -7, -9, -13, -19, -28 and TNF-α did not differ between UC patients with presence of intestinal MAP DNA compared to those without. MMP-2, MMP-9 and MMP-13 were significantly decreased in UC patients receiving corticosteroids. Conclusions The presence of intestinal MAP specific DNA is not associated with altered MMP expression in UC in vivo. Corticosteroids are associated with increased detection of intestinal MAP DNA and decreased expression of certain MMPs. Frequent detection of MAP DNA in healthy controls might be attributable to the wide environmental distribution of MAP and its presence in the food-chain.

  20. Genome-wide association analysis and genomic prediction of Mycobacterium avium subspecies paratuberculosis infection in US Jersey cattle.

    Science.gov (United States)

    Zare, Yalda; Shook, George E; Collins, Michael T; Kirkpatrick, Brian W

    2014-01-01

    Paratuberculosis (Johne's disease), an enteric disorder in ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP), causes economic losses in excess of $200 million annually to the US dairy industry. To identify genomic regions underlying susceptibility to MAP infection in Jersey cattle, a case-control genome-wide association study (GWAS) was performed. Blood and fecal samples were collected from ∼ 5,000 mature cows in 30 commercial Jersey herds from across the US. Discovery data consisted of 450 cases and 439 controls genotyped with the Illumina BovineSNP50 BeadChip. Cases were animals with positive ELISA and fecal culture (FC) results. Controls were animals negative to both ELISA and FC tests that matched cases on birth date and herd. Validation data consisted of 180 animals including 90 cases (positive to FC) and 90 controls (negative to ELISA and FC), selected from discovery herds and genotyped by Illumina BovineLD BeadChip (∼ 7K SNPs). Two analytical approaches were used: single-marker GWAS using the GRAMMAR-GC method and Bayesian variable selection (Bayes C) using GenSel software. GRAMMAR-GC identified one SNP on BTA7 at 68 megabases (Mb) surpassing a significance threshold of 5 × 10(-5). ARS-BFGL-NGS-11887 on BTA23 (27.7 Mb) accounted for the highest percentage of genetic variance (3.3%) in the Bayes C analysis. SNPs identified in common by GRAMMAR-GC and Bayes C in both discovery and combined data were mapped to BTA23 (27, 29 and 44 Mb), 3 (100, 101, 106 and 107 Mb) and 17 (57 Mb). Correspondence between results of GRAMMAR-GC and Bayes C was high (70-80% of most significant SNPs in common). These SNPs could potentially be associated with causal variants underlying susceptibility to MAP infection in Jersey cattle. Predictive performance of the model developed by Bayes C for prediction of infection status of animals in validation set was low (55% probability of correct ranking of paired case and control samples). PMID:24523889

  1. The other way around: probiotic Lactobacillus acidophilus NP51 restrict progression of Mycobacterium avium subspecies paratuberculosis (MAP) infection in Balb/c mice via activiation of CD8 alpha+ immune cell-mediated immunity

    Science.gov (United States)

    The objective of this study was to examine the immune-modulating effects of feeding a novel probiotic Lactobacillus acidophilus strain NP51 to specific pathogen-free Balb/c mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne’s disease (JD) in rumi...

  2. The other way around: Probiotic lactobacillus acidophilus NP51 restricts progression of Mycobacterium avium subspecies paratuberculosis (MAP) infection in Balb/c mice through activation of CD8+ T cell-mediated immunity

    Science.gov (United States)

    The objective of this study was to examine immune effects of feeding novel probiotic Lactobacillus acidophilus strain NP51 to specific pathogen-free Balb/c mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne’s disease (JD). We hypothesized that fe...

  3. Risk factors for the introduction and within-herd transmission of Mycobacterium avium subspecies paratuberculosis (MAP infection on 59 Irish dairy herds

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    Cashman W

    2008-07-01

    Full Text Available Abstract Since 1994, Irish cattle have been exposed to greater risks of acquiring Mycobacterium avium subspecies paratuberculosis (MAP infection as a consequence of the importation of over 70,000 animals from continental Europe. In recent years, there has been an increase in the number of reported clinical cases of paratuberculosis in Ireland. This study examines the prevalence of factors that promote the introduction and within-herd transmission of Mycobacterium avium subspecies paratuberculosis (MAP on selected Irish dairy farms in the Cork region, and the association between these factors and the results of MAP screening tests on milk sock filter residue (MFR. A total of 59 dairy farms, selected using non-random methods but apparently free of endemic paratuberculosis, were enrolled into the study. A questionnaire was used to collect data about risk factors for MAP introduction and transmission. The MFR was assessed on six occasions over 24 months for the presence of MAP, using culture and immunomagnetic separation prior to polymerase chain reaction (IMS-PCR. Furthermore, blood samples from all entire male and female animals over one year of age in 20 herds were tested by ELISA. Eighteen (31% farms had operated as closed herds since 1994, 28 (47% had purchased from multiple sources and 14 (24% had either direct or indirect (progeny contact with imported animals. Milk and colostrum were mixed on 51% of farms, while 88% of farms fed pooled milk. Thirty (51% herds tested negative to MFR culture and IMS-PCR, 12 (20% were MFR culture positive, 26 (44% were IMS-PCR positive and seven (12% were both culture and IMS-PCR positive. The probability of a positive MFR culture was significantly associated with reduced attendance at calving, and with increased use of individual calf pens and increased (but not significantly if mulitiple suckling was practised. There was poor agreement between MFR culture and MFR IMS-PCR results, but moderate agreement

  4. Linking chronic infection and autoimmune diseases: Mycobacterium avium subspecies paratuberculosis, SLC11A1 polymorphisms and type-1 diabetes mellitus.

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    Daniela Paccagnini

    Full Text Available BACKGROUND: The etiology of type 1 diabetes mellitus (T1DM is still unknown; numerous studies are performed to unravel the environmental factors involved in triggering the disease. SLC11A1 is a membrane transporter that is expressed in late endosomes of antigen presenting cells involved in the immunopathogenic events leading to T1DM. Mycobacterium avium subsp. paratuberculosis (MAP has been reported to be a possible trigger in the development of T1DM. METHODOLOGY/PRINCIPAL FINDINGS: Fifty nine T1DM patients and 79 healthy controls were genotyped for 9 polymorphisms of SLC11A1 gene, and screened for the presence of MAP by PCR. Differences in genotype frequency were evaluated for both T1DM patients and controls. We found a polymorphism in the SLC11A1 gene (274C/T associated to type 1 diabetic patients and not to controls. The presence of MAP DNA was also significantly associated with T1DM patients and not with controls. CONCLUSIONS/SIGNIFICANCE: The 274C/T SCL11A1 polymorphism was found to be associated with T1DM as well as the presence of MAP DNA in blood. Since MAP persists within macrophages and it is also processed by dendritic cells, further studies are necessary to evaluate if mutant forms of SLC11A1 alter the processing or presentation of MAP antigens triggering thereby an autoimmune response in T1DM patients.

  5. Johne's disease: reliability of environmental sampling to characterize Mycobacterium avium subspecies paratuberculosis (MAP) infection in beef cow-calf herds.

    Science.gov (United States)

    Klawonn, W; Einax, E; Pützschel, R; Schmidt, M; Donat, K

    2016-08-01

    Environmental samples are considered to be a cost-effective method of identifying Mycobacterium avium subsp. paratuberculosis (MAP)-positive dairy herds, but evidence for beef cow-calf herds is weak. This study aims at evaluating this approach in a total of 20 German herds that were characterized by individual faecal samples (n = 2545) of all cows. For 14 MAP-positive herds having at least one MAP-positive animal, the within-herd prevalence was calculated from concurrent individual faecal culture-based testing. Six herds certified as 'MAP free' based on the negative results of previous years served as MAP-negative controls. On average, six environmental samples were taken at the end of winter from areas with high cow traffic and tested for MAP by faecal culture. According to the environmental samples, nine (64·3%) out of the 14 MAP-positive cow-calf herds were infected. The percentage of positive environmental samples and the apparent within-herd prevalence (Spearman's P = 0·73, P test results (positive and negative) and the herd's status based on individual testing (Fisher's exact test, P = 0·014) showed a positive association. Considering limitations in low-prevalence herds, MAP-positive beef cow-calf herds are detectable by environmental samples in temperate climate zones. PMID:27094619

  6. Mycobacterium avium Subspecies paratuberculosis and Bovine Leukemia Virus Seroprevalence and Associated Risk Factors in Commercial Dairy and Beef Cattle in Northern and Northeastern China

    Science.gov (United States)

    Sun, Wu-Wen; Lv, Wen-Fa; Cong, Wei; Meng, Qing-Feng; Wang, Chun-Feng; Shan, Xiao-Feng; Qian, Ai-Dong

    2015-01-01

    Mycobacterium avium subspecies paratuberculosis (MAP) and bovine leukemia virus (BLV) are important pathogens, commonly responsible for economical loss to cattle farms all over the world, yet their epidemiology in commercial dairy and beef cattle in China is still unknown. Thus, from September 2013 to December 2014, a large-scale seroprevalence study was conducted to determine the seroprevalence and identify herd-level risk factors associated with MAP and BLV infection. The source sample was 3674 cattle from 113 herds in northern and northeastern China. Antibodies against MAP and BLV were detected using ELISA tests. At animal-level, the seroprevalence of antibodies against MAP and BLV was 11.79% (433/3674) and 18.29% (672/3674), respectively. At herd-level, the seroprevalence of antibodies against MAP and BLV was 20.35% and 21.24% (24/113), respectively. Herd size was identified to be associated with MAP infection while herd size and presence of cattle introduced from other farms were significantly associated with BLV infection. Further research is needed to confirm these findings and improve the knowledge of the epidemiology of these two pathogens in these regions and elsewhere in China. PMID:26504798

  7. Evaluation of "Indigenous Absorbed ELISA Kit" for the Estimation of Seroprevalence of Mycobacterium avium Subspecies paratuberculosis Antibodies in Human Beings in North India.

    Science.gov (United States)

    Singh, A V; Singh, S V; Verma, D K; Yadav, R; Singh, P K; Sohal, J S

    2011-01-01

    In present pilot study aimed to estimate, presence of Mycobacterium avium subspecies paratuberculosis (MAP) antibodies in the human serum samples originating from North India using "Indigenous absorbed ELISA kit" (ELISA kit). The phase I, "ELISA kit" was optimized using protoplasmic antigen from native isolate of MAP "Indian Bison type" recovered from the biopsies of Crohn's disease patients. The phase II, sensitivity and specificity of the kit were estimated as 40.0 and 83.3%, respectively, when evaluated in 40 human serum samples (5 Crohn's disease and 22 ulcerative colitis patients and 13 healthy human subjects) with defined MAP status with respect to stool culture. Seroprevalence of MAP antibodies was higher in CD patients (80.0%) as compared to ulcerative colitis patients (4.5%) and normal human subjects (15.3%). The phase III, seroprevalence of MAP antibodies was estimated as 23.4%, on the basis of the screening of 452 human serum samples (without history) from different geographical regions of North India. Region-wise, 34.0, 33.3, 32.8, 25.0, 23.0, 17.7, and 12.5% samples were positive from the states of Punjab, Uttarakhand, New Delhi, Himachal Pradesh, Haryana, Uttar Pradesh and Jammu and Kashmir, respectively. Study reported moderately higher presence of MAP antibodies in human population, which necessitates programs to reduce the bioburden of MAP in the environment and in animal population. PMID:23738105

  8. The Features of Fecal and Ileal Mucosa-Associated Microbiota in Dairy Calves during Early Infection with Mycobacterium avium Subspecies paratuberculosis.

    Science.gov (United States)

    Derakhshani, Hooman; De Buck, Jeroen; Mortier, Rienske; Barkema, Herman W; Krause, Denis O; Khafipour, Ehsan

    2016-01-01

    Current diagnostic tests for Johne's disease (JD), a chronic granulomatous inflammation of the gastrointestinal tract of ruminants caused by Mycobacterium avium subspecies paratuberculosis (MAP), lack the sensitivity to identify infected animals at early (asymptomatic) stages of the disease. The objective was to determine the pattern of MAP-associated dysbiosis of intestinal microbiota as a potential biomarker for early detection of infected cattle. To that end, genomic DNA was extracted from ileal mucosa and fecal samples collected from 28 MAP-positive and five control calves. High-throughput Illumina sequencing of the V4 hypervariable region of the 16S rRNA gene was used for community profiling of ileal mucosa-associated (MAM) or fecal microbiota. The PERMANOVA analysis of unweighted UniFrac distances revealed distinct clustering of ileal MAM (P = 0.049) and fecal microbiota (P = 0.068) in MAP-infected vs. control cattle. Microbiota profile of MAP-infected animals was further investigated by linear discriminant analysis effective size (LEfSe); several bacterial taxa within the phylum Proteobacteria were overrepresented in ileal MAM of control calves. Moreover, based on reconstructed metagenomes (PICRUSt) of ileal MAM, functional pathways associated with MAP infection were inferred. Enrichment of lysine and histidine metabolism pathways, and underrepresentation of glutathione metabolism and leucine and isoleucine degradation pathways in MAP-infected calves suggested potential contributions of ileal MAM in development of intestinal inflammation. Finally, simultaneous overrepresentation of families Planococcaceae and Paraprevotellaceae, as well as underrepresentation of genera Faecalibacterium and Akkermansia in the fecal microbiota of infected cattle, served as potential biomarker for identifying infected cattle during subclinical stages of JD. Collectively, based on compositional and functional shifts in intestinal microbiota of infected cattle, we inferred that

  9. Detection of Mycobacterium avium subspecies paratuberculosis specific IS900 insertion sequences in bulk-tank milk samples obtained from different regions throughout Switzerland

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    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.

  10. Mycobacterium avium Subspecies paratuberculosis Infection in Cases of Irritable Bowel Syndrome and Comparison with Crohn's Disease and Johne's Disease: Common Neural and Immune Pathogenicities▿

    OpenAIRE

    Scanu, Antonio M.; Tim J Bull; Cannas, Sara; Sanderson, Jeremy D; Sechi, Leonardo A.; Dettori, Giuseppe; Zanetti, Stefania; Hermon-Taylor, John

    2007-01-01

    Mycobacterium avium subsp. paratuberculosis causes Johne's disease, a systemic infection and chronic inflammation of the intestine that affects many species, including primates. Infection is widespread in livestock, and human populations are exposed. Johne's disease is associated with immune dysregulation, with involvement of the enteric nervous system overlapping with features of irritable bowel syndrome in humans. The present study was designed to look for an association between Mycobacteri...

  11. 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.

  12. Successful treatment of asymptomatic or clinically terminal bovine Mycobacterium avium subspecies paratuberculosis infection (Johne's disease) with the bacterium Dietzia used as a probiotic alone or in combination with dexamethasone: Adaption to chronic human diarrheal diseases.

    Science.gov (United States)

    Click, Robert E

    2011-01-01

    A naturally occurring gastrointestinal disease, primarily of ruminants (Johne disease), is a chronic debilitating disease that is caused by Mycobacterium avium subspecies paratuberculosis (MAP). MAP infection occurs primarily in utero and in newborns. Outside our Dietzia probiotic treatment, there are no preventive/curative therapies for bovine paratuberculosis. Interestingly, MAP is at the center of controversy as to its role in (cause of) Crohn disease (CD) and more recently, its role in diabetes, ulcerative colitis, and irritable bowel syndrome (IBS); the latter two, like CD, are considered to be a result of chronic intestinal inflammation. Treatments, both conventional and biologic agents, which induce and maintain remission are directed at curtailing processes that are an intricate part of inflammation. Most possess side effects of varying severity, lose therapeutic value, and more importantly, none routinely result in prevention and/or cures. Based on (a) similarities of Johne disease and Crohn disease, (b) a report that Dietzia inhibited growth of MAP under specific culture conditions, and (c) findings that Dietzia when used as a probiotic, (i) was therapeutic for adult bovine paratuberculosis, and (ii) prevented development of disease in MAP-infected calves, the goal of the present investigations was to design protocols that have applicability for IBD patients. Dietzia was found safe for cattle of all ages and for normal and immunodeficient mice. The results strongly warrant clinical evaluation as a probiotic, in combination with/without dexamethasone. PMID:21460639

  13. Lack of association between the occurrence of Crohn's disease and occupational exposure to dairy and beef cattle herds infected with Mycobacterium avium subspecies paratuberculosis.

    Science.gov (United States)

    Qual, D A; Kaneene, J B; Varty, T J; Miller, R; Thoen, C O

    2010-06-01

    A cross-sectional survey was conducted to identify associations between Crohn's disease (CD) and Mycobacterium avium ssp. paratuberculosis (Map) exposure. A questionnaire was used to collect information on exposure to cattle infected with Map, and personal and family history of CD in dairy and beef cattle producers with and without Map-infected herds, and in veterinarians who did or did not have contact with Map-infected herds. Cases of CD were selected from respondents and matched 1:4 with controls on occupation, age, and sex. Multivariable conditional logistic regression was used to assess associations between Map exposure and CD. There were 3 cases of CD in 702 producers and 4 cases in 774 veterinarians, yielding a prevalence of 0.47%. No association was found between exposure to JD and CD in any phase of the analysis. However, the number of cases of CD is not large and limits the power to detect important differences. PMID:20494145

  14. Dam Mycobacterium avium subspecies paratuberculosis (MAP) infection status does not predetermine calves for future shedding when raised in a contaminated environment: a cohort study.

    Science.gov (United States)

    Eisenberg, Susanne W F; Rutten, Victor P M G; Koets, Ad 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 presence of MAP DNA and antibody in colostrum and that of DNA in faeces and the environment with subsequent MAP shedding of their daughters. A cohort of 117 dam-daughter pairs giving birth/being born on eight commercial dairy farms with endemic paratuberculosis was followed where colostrum, faecal and environmental samples (dust) were analysed for the presence of MAP using an IS900 real-time PCR. Antibodies in colostrum were measured by ELISA. Analysis of dust samples showed that on all farms environmental MAP exposure occurred continuously. In significantly more colostrum samples (48%) MAP DNA was detected compared to faecal samples (37%). MAP specific antibodies were present in 34% of the colostrum samples. In total MAP DNA was present in faecal samples of 41% of the daughters at least once during the sampling period. The association between faecal shedding in the offspring and the dam MAP status defined by MAP PCR on colostrum, MAP PCR on faeces or ELISA on colostrum was determined by an exact cox regression analysis for discrete data. The model indicated that the hazard for faecal shedding in daughters born to MAP positive dams was not significantly different compared to daughters born to MAP negative dams. When born to a dam with DNA positive faeces the HR was 1.05 (CI 0.6; 1.8) and with DNA positive colostrum the HR was 1.17 (CI 0.6; 2.3). When dam status was defined by a combination of both PCR outcomes (faeces and colostrum) and the ELISA outcome the HR was 1.26 (CI 0.9; 1.9). Therefore, this study indicates that neither the presence of MAP DNA in colostrum, MAP DNA in faeces nor the presence of MAP antibodies in colostrum of the dam significantly influences the hazard of

  15. Detection of Mycobacterium avium subspecies paratuberculosis in tie-stall dairy herds using a standardized environmental sampling technique and targeted pooled samples.

    Science.gov (United States)

    Arango-Sabogal, Juan C; Côté, Geneviève; Paré, Julie; Labrecque, Olivia; Roy, Jean-Philippe; Buczinski, Sébastien; Doré, Elizabeth; Fairbrother, Julie H; Bissonnette, Nathalie; Wellemans, Vincent; Fecteau, Gilles

    2016-07-01

    Mycobacterium avium ssp. paratuberculosis (MAP) is the etiologic agent of Johne's disease, a chronic contagious enteritis of ruminants that causes major economic losses. Several studies, most involving large free-stall herds, have found environmental sampling to be a suitable method for detecting MAP-infected herds. In eastern Canada, where small tie-stall herds are predominant, certain conditions and management practices may influence the survival and transmission of MAP and recovery (isolation). Our objective was to estimate the performance of a standardized environmental and targeted pooled sampling technique for the detection of MAP-infected tie-stall dairy herds. Twenty-four farms (19 MAP-infected and 5 non-infected) were enrolled, but only 20 were visited twice in the same year, to collect 7 environmental samples and 2 pooled samples (sick cows and cows with poor body condition). Concurrent individual sampling of all adult cows in the herds was also carried out. Isolation of MAP was achieved using the MGIT Para TB culture media and the BACTEC 960 detection system. Overall, MAP was isolated in 7% of the environmental cultures. The sensitivity of the environmental culture was 44% [95% confidence interval (CI): 20% to 70%] when combining results from 2 different herd visits and 32% (95% CI: 13% to 57%) when results from only 1 random herd visit were used. The best sampling strategy was to combine samples from the manure pit, gutter, sick cows, and cows with poor body condition. The standardized environmental sampling technique and the targeted pooled samples presented in this study is an alternative sampling strategy to costly individual cultures for detecting MAP-infected tie-stall dairies. Repeated samplings may improve the detection of MAP-infected herds. PMID:27408329

  16. Detection of Mycobacterium avium subspecies paratuberculosis in patients with Crohn's disease is unrelated to the presence of single nucleotide polymorphisms rs2241880 (ATG16L1) and rs10045431 (IL12B).

    Science.gov (United States)

    Dalton, James P; Desmond, Alan; Shanahan, Fergus; Hill, Colin

    2014-06-01

    Mycobacterium avium subspecies paratuberculosis (MAP) has been controversially linked with Crohn's disease (CD). Detection of MAP in CD has been highly variable, and one explanation might be the genetic heterogeneity of this syndrome. Many of the single nucleotide polymorphisms (SNPs) linked with CD are contained within genes that are associated with bacterial handling in general, and some are specifically implicated in susceptibility to mycobacterial disease. We tested a cohort of IBD patients (n = 149) to determine whether the presence of MAP was associated with a selection of these SNPs. Blood samples from CD patients (n = 84), ulcerative colitis (UC, n = 65) patients and healthy controls (n = 55) were examined for the presence of MAP and SNPs in ATG16L1, IL12B, NOD2/CARD15, NKx2-3, IL23R and IRGM. Statistical analysis was then used to determine whether there was any association between the presence of MAP and these SNPs. MAP, rs2241880 (ATG16L1) and rs10045431 (IL12B) were found to be significantly associated with CD. The presence of MAP was not related to the status of the SNPs in ATG16L1 or IL12B. We have found no evidence for the contribution of these SNPs to the presence of MAP in CD patients. PMID:24522266

  17. MYCOBACTERIUM AVIUM SUSP. PARATUBERCULOSIS IN DAIRY PRODUCTION

    Directory of Open Access Journals (Sweden)

    G. Marchetti

    2012-08-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (MAP is the etiologic agent of paratuberculosis. The disease affects cows and other ruminants and causes high economic losses, mainly for dairy production. MAP may also have a role in the development of Crohn’s disease in humans. Infected animals shed viable MAP with milk and faeces and humans may assume MAP via the consumption of contaminated milk and dairy products. Current methods of milk pasteurization are not sufficient to kill all MAP cells present in milk and MAP has been found in raw or pasteurized milk and isolated from cheese. The aim of this paper is to review the current knowledge about MAP in dairy production. We analyzed studies on milk contamination, effect of pasteurization and methods for identification of MAP that can be applied to dairy products.

  18. Is Mycobacterium avium subsp. paratuberculosis, the cause of Johne's disease in animals, a good candidate for Crohn's disease in man?

    Science.gov (United States)

    Singh, A V; Singh, S V; Singh, P K; Sohal, J S

    2010-03-01

    Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne's disease or paratuberculosis, a gastro intestinal inflammatory condition in ruminants and other animals, which is similar to Crohn's disease (CD) that occurs in man. The role of MAP in the causation of CD has been under intense investigation in the last few decades. This review summarizes the status of MAP in animals and the food chain and its association with CD in man. PMID:20443099

  19. 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).

  20. 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...

  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. 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). Mice were randomized to ten treatment groups; sentinels, control, heat-killed MAP, viable MAP, heat-killed NP51, viable ...

  4. TLR4, IL10RA, and NOD2 mutation in paediatric Crohn's disease patients: an association with Mycobacterium avium subspecies paratuberculosis and TLR4 and IL10RA expression.

    Science.gov (United States)

    Wagner, Josef; Skinner, Narelle A; Catto-Smith, Anthony G; Cameron, Donald J S; Michalski, Wojtek P; Visvanathan, Kumar; Kirkwood, Carl D

    2013-08-01

    Mycobacterium avium subspecies paratuberculosis (MAP) has been implicated in the pathogenesis of Crohn's disease (CD). The role of CD susceptibility genes in association with these microbes is not known. Sixty-two early onset paediatric CD patients and 46 controls with known MAP status were analysed for an association with 34 single nucleotide polymorphisms (SNPs) from 18 CD susceptibility genes. Functional studies on peripheral blood mononuclear cells (PBMCs) were conducted on 17 CD patients with known CD mutations to assess IL-6, IL-10, and TNF-α expression upon stimulation with MAP precipitated protein derivative (PPD) and lipopolysaccharide (LPS). In addition, surface expression of IL10R and TLR4 on resting B cells, NK cells, T cells, and monocytes was assessed. A mutation in TLR4 (rs4986790) and IL10RA (rs22291130) was significantly associated with MAP-positive CD patients compared to MAP-negative CD patients (27.6 vs. 6.1 %, p = 0.021, and 62.1 vs. 33.3 %, p = 0.024, respectively). PPD and LPS significantly increased IL-6, IL-10, and TNF-α production in PBMCs. IL-10 and TNF-α production were significantly lower in a subgroup of CD patients (5/12) with a known NOD2 mutation. Receptor for IL-10 was significantly higher expressed on NK cells (CD56low) and on NK T cells harbouring a NOD2 mutations compared to wildtype cells (p = 0.031 and 0.005, respectively). TLR4 was significantly higher expressed on NK cells (CD56high) harbouring a NOD2 mutations compared to wildtype cells (p = 0.038). PMID:23455702

  5. LCD array and IS900 efficiency in relation to traditional diagnostic techniques for diagnosis of Mycobacterium avium subspecies paratuberculosis in cattle in Egypt.

    Science.gov (United States)

    ElSayed, Mohamed Sabry Abd ElRaheam

    2014-06-01

    This study aimed to compare traditional tests (Johnin test, fecal staining and fecal culture) with advanced laboratory tests (ELISA, LCD array and IS900 PCR) for detection of Johne's disease. A total of 365 Holstein-Friesian dairy cattle (40 express profuse diarrhea unresponsive to treatment and 325 contacting them) tested with Johnin test, blood collected for ELISA and fecal samples for fecal staining as well as fecal culture, application of LCD array and PCR using IS900 on DNA extracted from Mycobacterium paratuberculosis bacilli (from feces and culture). Johnin test was 40/40 (100%) and 25/325 (7.69%), fecal staining was 13 (37.1%) and 2 (50%), ELISA was 35/40 (87.5%) and 4/25 (16%) for clinical cattle and apparently healthy contacting them respectively. Isolation was 12/13 (92.3%) of the (Johnin test +ve, ELISA +ve and Acid Fast Bacilli +ve) from the clinically positive cattle and 1/2 (50%) of the (Johnin test +ve, ELISA +ve and Acid Fast Bacilli +ve) from apparently healthy contacting them while LCD array and IS900 gave 100% confirming the isolation results. In conclusion, LCD array depending on 16S RNA and DNA hybridization with specific probes for detection of M. paratuberculosis are fast, sensitive and labor-saving when combined with IS900. PMID:26786331

  6. 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...... comprising the one or more immunogenic polypeptides, is provided for prevention or treatment of infection with a Mycobacterium species, e.g. Mycobacterium avium subsp. paratuberculosis....

  7. Mycobacterium avium subsp. paratuberculosis infection, immunology and pathology of livestock

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) infection in ruminants leads to a chronic and progressive enteric disease (Johne’s disease) that results in loss of intestinal function, poor body condition, and eventual death. Transmission is primarily through a fecal-oral route in neonates but con...

  8. 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

    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...

  9. Development and Characterization of Monoclonal Antibodies and Aptamers Against Major Antigens of Mycobacterium avium subsp. paratuberculosis

    Science.gov (United States)

    Specific antibodies, available in unlimited quantities, have not been produced against Mycobacterium avium subsp. paratuberculosis, the bacterium that causes Johne’s disease (JD). To fill this gap in JD research, monoclonal antibodies (mAbs) against M. avium subsp. paratuberculosis were produced fr...

  10. Culture Phenotypes of Genomically and Geographically Diverse Mycobacterium avium subsp. paratuberculosis Isolates from Different Hosts▿

    OpenAIRE

    Whittington, Richard J.; Marsh, Ian B; Saunders, Vanessa; Grant, Irene R.; Juste, Ramon; Sevilla, Iker A; Manning, Elizabeth J. B.; Whitlock, Robert H.

    2011-01-01

    Mycobacterium avium subsp. paratuberculosis causes paratuberculosis (Johne's disease) in ruminants in most countries. Historical data suggest substantial differences in culturability of M. avium subsp. paratuberculosis isolates from small ruminants and cattle; however, a systematic comparison of culture media and isolates from different countries and hosts has not been undertaken. Here, 35 field isolates from the United States, Spain, Northern Ireland, and Australia were propagated in Bactec ...

  11. Assessing the inactivation of Mycobacterium avium subsp. paratuberculosis during composting of livestock carcasses.

    Science.gov (United States)

    Tkachuk, Victoria L; Krause, Denis O; McAllister, Tim A; Buckley, Katherine E; Reuter, Tim; Hendrick, Steve; Ominski, Kim H

    2013-05-01

    Mycobacterium avium subsp. paratuberculosis causes Johne's disease (JD) in ruminants, with substantial economic impacts on the cattle industry. Johne's disease is known for its long latency period, and difficulties in diagnosis are due to insensitivities of current detection methods. Eradication is challenging as M. avium subsp. paratuberculosis can survive for extended periods within the environment, resulting in new infections in naïve animals (W. Xu et al., J. Environ. Qual. 38:437-450, 2009). This study explored the use of a biosecure, static composting structure to inactivate M. avium subsp. paratuberculosis. Mycobacterium smegmatis was also assessed as a surrogate for M. avium subsp. paratuberculosis. Two structures were constructed to hold three cattle carcasses each. Naturally infected tissues and ground beef inoculated with laboratory-cultured M. avium subsp. paratuberculosis and M. smegmatis were placed in nylon and plastic bags to determine effects of temperature and compost environment on viability over 250 days. After removal, samples were cultured and growth of both organisms was assessed after 12 weeks. After 250 days, M. avium subsp. paratuberculosis was still detectable by PCR, while M. smegmatis was not detected after 67 days of composting. Furthermore, M. avium subsp. paratuberculosis remained viable in both implanted nylon and plastic bags over the composting period. As the compost never reached a homogenous thermophilic (55 to 65°C) state throughout each structure, an in vitro experiment was conducted to examine viability of M. avium subsp. paratuberculosis after exposure to 80°C for 90 days. Naturally infected lymph tissues were mixed with and without compost. After 90 days, M. avium subsp. paratuberculosis remained viable despite exposure to temperatures typically higher than that achieved in compost. In conclusion, it is unlikely composting can be used as a means of inactivating M. avium subsp. paratuberculosis associated with cattle

  12. Determination of genotypic diversity of Mycobacterium avium subspecies from human and animal origins by mycobacterial interspersed repetitive-unit-variable-number tandem-repeat and IS1311 restriction fragment length polymorphism typing methods.

    Science.gov (United States)

    Radomski, Nicolas; Thibault, Virginie C; Karoui, Claudine; de Cruz, Krystel; Cochard, Thierry; Gutiérrez, Cristina; Supply, Philip; Biet, Frank; Boschiroli, María Laura

    2010-04-01

    Members of the Mycobacterium avium complex (MAC) are ubiquitous bacteria that can be found in water, food, and other environmental samples and are considered opportunistic pathogens for numerous animal species, mainly birds and pigs, as well as for humans. We have recently demonstrated the usefulness of a PCR-based mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) typing for the molecular characterization of M. avium subsp. paratuberculosis and M. avium strains exclusively isolated from AIDS patients. In the present study we extended our analysis, based on eight MIRU-VNTR markers, to a strain collection comprehensively comprising the other M. avium subspecies, including M. avium subsp. avium, M. avium subsp. hominissuis, and M. avium subsp. silvaticum, isolated from numerous animal species, HIV-positive and HIV-negative humans, and environmental sources. All strains were fully typeable, with the discriminatory index being 0.885, which is almost equal to that obtained by IS1311 restriction fragment length polymorphism (RFLP) typing as a reference. In contrast to IS1311 RFLP typing, MIRU-VNTR typing was able to further discriminate M. avium subsp. avium strains. MIRU-VNTR alleles strongly associated with or specific for M. avium subspecies were detected in several markers. Moreover, the MIRU-VNTR typing-based results were consistent with a scenario of the independent evolution of M. avium subsp. avium/M. avium subsp. silvaticum and M. avium subsp. paratuberculosis from M. avium subsp. hominissuis, previously proposed on the basis of multilocus sequence analysis. MIRU-VNTR typing therefore appears to be a convenient typing method capable of distinguishing the three main subspecies and strains of the complex and providing new epidemiological knowledge on MAC. PMID:20107094

  13. 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...... animals. In milk from five cows (all faeces culture positive), we cultivated a few colonies of M. paratuberculosis (<100 CFU per ml). Milk samples from two cows were PCR positive (both animals were faeces culture positive, and one cow was milk culture positive). One cow was culture negative on intestinal...

  14. Assessing the Inactivation of Mycobacterium avium subsp. paratuberculosis during Composting of Livestock Carcasses

    OpenAIRE

    Tkachuk, Victoria L.; Denis O Krause; McAllister, Tim A.; Buckley, Katherine E.; Reuter, Tim; Hendrick, Steve; Ominski, Kim H.

    2013-01-01

    Mycobacterium avium subsp. paratuberculosis causes Johne's disease (JD) in ruminants, with substantial economic impacts on the cattle industry. Johne's disease is known for its long latency period, and difficulties in diagnosis are due to insensitivities of current detection methods. Eradication is challenging as M. avium subsp. paratuberculosis can survive for extended periods within the environment, resulting in new infections in naïve animals (W. Xu et al., J. Environ. Qual. 38:437-450, 20...

  15. Rapid Expression of Mycobacterium avium subsp. paratuberculosis Recombinant Proteins for Antigen Discovery▿

    OpenAIRE

    Li, Lingling; Munir, Shirin; Bannantine, John P.; Sreevatsan, Srinand; Kanjilal, Sagarika; Kapur, Vivek

    2006-01-01

    Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease, a chronic granulomatous enteritis of ruminants and other species. Detection of infection in animals is hampered by the lack of sensitive and specific diagnostic assays. We describe here an approach that utilizes translationally active PCR fragments for the rapid in vitro transcription and translation of recombinant proteins for antigen discovery in M. avium subsp. paratuberculosis. The investigations showed...

  16. Different Mycobacterium avium subsp. paratuberculosis MIRU-VNTR patterns coexist within cattle herds

    NARCIS (Netherlands)

    Hulzen, van K.J.E.; Heuven, H.C.M.; Nielen, M.; Hoeboer, J.; Santema, W.J.; Koets, A.P.

    2011-01-01

    A better understanding of the biodiversity of Mycobacterium avium subsp. paratuberculosis (MAP) offers more insight in the epidemiology of paratuberculosis and therefore may contribute to the control of the disease. The aim of this study was to investigate the genetic diversity in bovine MAP isolate

  17. 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

    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...

  18. 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

    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-negative on...

  19. Putative in vitro expressed gene fragments unique to Mycobacterium avium subspecies para tuberculosis

    DEFF Research Database (Denmark)

    Nielsen, Kirstine Klitgaard; Ahrens, Peter

    2002-01-01

    By a suppression subtractive hybridization based method, nine novel Mycobacterium avium subsp. para tuberculosis (M. para tuberculosis) fragments of between 318 and 596 bp have been identified and characterized. Database search revealed little or no similarity with other mycobacteria. The...... uniqueness and diagnostic potential of seven of these fragments in relation to M. paratuberculosis closest relative Mycobacterium avium subsp. avium (M. avium) was confirmed by species-specific PCR and Southern blot. Furthermore, RT-PCR indicated that eight of the nine fragments originate from areas of the...

  20. Immunological findings associated with Argentinean strains of Mycobacterium avium subsp. paratuberculosis in bovine models.

    Science.gov (United States)

    Colavecchia, Silvia B; Fernández, Bárbara; Jolly, Ana; Minatel, Leonardo; Hajos, Silvia E; Paolicchi, Fernando A; Mundo, Silvia L

    2016-08-01

    Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of ruminant paratuberculosis. The aim of this study was to evaluate the biological behavior of different Argentinean strains of MAP in two bovine infection models: macrophage (in vitro) and calf (in vivo) through the evaluation of early immune responses at the peripheral and local levels. Two MAP strains (A and C) were selected taking into account the different patterns of TNF-α and IL-10 secretion displayed by infected bovine macrophages in vitro. Two groups of calves were infected with 250mg of total wet weight live MAP: strain A infected group (MA, n=3), strain C infected group (MC, n=2). Another group of animals was mock-infected (MI, n=3). Infection was confirmed by MAP culture of feces and microscopic observation of granulomatous lesions in the gut tissue. All infected calves showed positive results in the DTH skin test. A significant increase in peripheral CD4CD25(+) cells in MC group on day 150 was detected. The specific cellular immune response developed allowed the identification of the infection as early as 30days in the MA group. However, the percentage of CD8CD25(+) cells was significantly increased on day 120 in MC group. Significant differences between groups in proliferation and cellular responses were also detected in ileocecal lymph node samples. In summary, the strains of MAP employed herein induced differential immune responses in peripheral cells, in the proliferative responses and in cell functionality at the local level. Our findings support the hypotheses that the in vitro behavior displayed by macrophages could be a tool to identify differences among MAP strains infecting bovines and that the host-pathogen interactions occurring upon infection are dependent on the strain of MAP involved. PMID:27138443

  1. Divergent immune responses to Mycobacterium avium subsp. paratuberculosis infection correlate with kinome responses at the site of intestinal infection.

    Science.gov (United States)

    Määttänen, Pekka; Trost, Brett; Scruten, Erin; Potter, Andrew; Kusalik, Anthony; Griebel, Philip; Napper, Scott

    2013-08-01

    Mycobacterium avium subsp. paratuberculosis is the causative agent of Johne's disease (JD) in cattle. M. avium subsp. paratuberculosis infects the gastrointestinal tract of calves, localizing and persisting primarily in the distal ileum. A high percentage of cattle exposed to M. avium subsp. paratuberculosis do not develop JD, but the mechanisms by which they resist infection are not understood. Here, we merge an established in vivo bovine intestinal segment model for M. avium subsp. paratuberculosis infection with bovine-specific peptide kinome arrays as a first step to understanding how infection influences host kinomic responses at the site of infection. Application of peptide arrays to in vivo tissue samples represents a critical and ambitious step in using this technology to understand host-pathogen interactions. Kinome analysis was performed on intestinal samples from 4 ileal segments subdivided into 10 separate compartments (6 M. avium subsp. paratuberculosis-infected compartments and 4 intra-animal controls) using bovine-specific peptide arrays. Kinome data sets clustered into two groups, suggesting unique binary responses to M. avium subsp. paratuberculosis. Similarly, two M. avium subsp. paratuberculosis-specific immune responses, characterized by different antibody, T cell proliferation, and gamma interferon (IFN-γ) responses, were also observed. Interestingly, the kinomic groupings segregated with the immune response groupings. Pathway and gene ontology analyses revealed that differences in innate immune and interleukin signaling and particular differences in the Wnt/β-catenin pathway distinguished the kinomic groupings. Collectively, kinome analysis of tissue samples offers insight into the complex cellular responses induced by M. avium subsp. paratuberculosis in the ileum and provides a novel method to understand mechanisms that alter the balance between cell-mediated and antibody responses to M. avium subsp. paratuberculosis infection. PMID

  2. Production and Characterization of Monoclonal Antibodies Against a Major Membrane Protein of Mycobacterium avium subsp. paratuberculosis

    Science.gov (United States)

    The Mycobacterium avium subsp. paratuberculosis 35-kDa major membrane protein (MMP) encoded by MAP2121c has been shown to play a role in invasion of epithelial cells and is an important membrane antigen recognized by cattle with Johne’s disease. In this study, purified recombinant MMP was used to p...

  3. Molecular Epidemiology of Mycobacterium avium subsp. paratuberculosis in a Longitudinal Study of Three Dairy Herds

    Science.gov (United States)

    The objective of this study was to evaluate whether cows that were low shedders of Mycobacterium avium subsp. paratuberculosis (MAP) were passive shedding animals or whether they were truly infected with MAP. We also evaluated whether these MAP-infected animals could have been infected as adults by ...

  4. Detection of Mycobacterium avium subsp. paratuberculosis in Drinking Water and Biofilms Using Quantitative PCR

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne’s disease in domestic animals and has been implicated in Crohn’s disease in humans. Cows infected with Johne’s disease shed large quantities of MAP into soil. Further, MAP has been isolated from surface water, is resi...

  5. From mouth to macrophage: mechanisms of innate immune subversion by Mycobacterium avium subsp. paratuberculosis

    Science.gov (United States)

    Johne’s disease (JD) is a chronic enteric infection of cattle caused by Mycobacterium avium subsp. paratuberculosis (MAP). The high economic cost and potential zoonotic threat of JD have driven efforts to develop tools and approaches to effectively manage this disease within livestock herds. Efforts...

  6. Optimization of hexadecylpyridinium chloride decontamination for culture of Mycobacterium avium subsp. paratuberculosis from milk

    Science.gov (United States)

    Cows in advanced stages of Johne’s disease shed Mycobacterium avium subsp. paratuberculosis (MAP) into both their milk and feces, allowing for transmission of the bacteria between animals. The objective of this study was to formulate an optimized protocol for the isolation of MAP from milk and colos...

  7. Mycobacterium avium subsp. paratuberculosis Fibronectin Attachment Protein Facilitates M-Cell Targeting and Invasion through a Fibronectin Bridge with Host Integrins

    OpenAIRE

    Secott, T. E.; Lin, T. L.; Wu, C. C.

    2004-01-01

    Efficient attachment and ingestion of Mycobacterium avium subsp. paratuberculosis by cultured epithelial cells requires the expression of a fibronectin (FN) attachment protein homologue (FAP-P) which mediates FN binding by M. avium subsp. paratuberculosis. Invasion of Peyer's patches by M. avium subsp. paratuberculosis occurs through M cells, which, unlike other intestinal epithelial cells, express integrins on their luminal faces. We sought to determine if the interaction between FAP-P of M....

  8. 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.

  9. Mediation of host immune responses after immunization of neonatal calves with a heat-killed Mycobacterium avium subsp. paratuberculosis vaccine

    Science.gov (United States)

    A major drawback of current whole-cell vaccines for Mycobacterium avium subsp. paratuberculosis(MAP) is the interference with diagnostic tests for bovine tuberculosis and paratuberculosis. The current study was designed to explore effects of immunization with a heat-killed whole cell vaccine (Mycop...

  10. A murine oral model for Mycobacterium avium subsp. paratuberculosis infection and immunomodulation with Lactobacillus casei ATCC 334

    OpenAIRE

    Cooney, Meagan A.; Steele, James L; Steinberg, Howard; Talaat, Adel M.

    2014-01-01

    Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) the causative agent of Johne's disease, is one of the most serious infectious diseases in dairy cattle worldwide. Due to the chronic nature of this disease and no feasible control strategy, it is essential to have an efficient animal model which is representative of the natural route of infection as well as a viable treatment option. In this report, we evaluated the effect of different doses of M. paratuberculosis in their abil...

  11. Proteome and Antigens of Mycobacterium avium subspecies Paratuberculosis

    Science.gov (United States)

    This chapter will review what is known about the proteome of this significant veterinary pathogen by discussing the use of various whole cell preparations and fractionated components thereof. With improvements in technology, many new modifications to the traditional ELISA assay have been introduced...

  12. No Holes Barred: Invasion of the Intestinal Mucosa by Mycobacterium avium subsp. paratuberculosis

    OpenAIRE

    Bannantine, John P; Bermudez, Luiz E.

    2013-01-01

    The infection biology of Mycobacterium avium subsp. paratuberculosis has recently crystallized, with added details surrounding intestinal invasion. The involvement of pathogen-derived effector proteins such as the major membrane protein, oxidoreductase, and fibronectin attachment proteins have been uncovered. Mutations constructed in this pathogen have also shed light on genes needed for invasion. The host cell types that are susceptible to invasion have been defined, along with their transcr...

  13. Host Responses to the Pathogen Mycobacterium avium subsp. paratuberculosis and Beneficial Microbes Exhibit Host Sex Specificity

    OpenAIRE

    Karunasena, Enusha; McMahon, K Wyatt; Chang, David; Brashears, Mindy M.

    2014-01-01

    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 th...

  14. Development of an F57 Sequence-Based Real-Time PCR Assay for Detection of Mycobacterium avium subsp. paratuberculosis in Milk

    OpenAIRE

    Tasara, T; Stephan, R.

    2005-01-01

    A light cycler-based real-time PCR (LC-PCR) assay that amplifies the F57 sequence of Mycobacterium avium subsp. paratuberculosis was developed. This assay also includes an internal amplification control template to monitor the amplification conditions in each reaction. The targeted F57 sequence element is unique for M.avium subsp. paratuberculosis and is not known to exist in any other bacterial species. The assay specificity was demonstrated by evaluation of 10 known M. avium subsp. paratube...

  15. Evidence for a Novel Gene Expression Program in Peripheral Blood Mononuclear Cells from Mycobacterium avium subsp. paratuberculosis-Infected Cattle

    OpenAIRE

    Coussens, Paul M.; Colvin, Christopher J.; Guilherme J. M. Rosa; Perez Laspiur, Juliana; Elftman, Michael D.

    2003-01-01

    A bovine-specific cDNA microarray system was used to compare gene expression profiles of peripheral blood mononuclear cells (PBMCs) from control uninfected (n = 4) and Johne's disease-positive (n = 6) Holstein cows. Microarray experiments were designed so that for each animal, a direct comparison was made between PBMCs stimulated in vitro with Mycobacterium avium subsp. paratuberculosis and PBMCs stimulated with phosphate-buffered saline (nil-stimulated PBMCs). As expected, M. avium subsp. pa...

  16. [Possible association between Mycobacterium avium subsp paratuberculosis infection and Crohn's disease].

    Science.gov (United States)

    Retamal, Patricio; Beltrán, Caroll; Abalos, Pedro; Quera, Rodrigo; Hermoso, Marcela

    2011-06-01

    Paratuberculosis is a chronic intestinal disease of animals caused by Mycobacterium avium subsp. paratuberculosis (MAP), which has some pathological features similar to Crohn's disease (CD) in humans. The presence of MAP in food for human consumption and in affected tissues of patients with CD has been detected. Therefore, a causal association between this microorganism and the disease in humans, has been postulated. However, several related studies have failed to confirm this hypothesis and the scientific acceptance of MAP as a zoonotic agent remains controversial. This review presents the main findings related to this issue, contrasting evidences for and against an association between MAP and CD. The need to promote national studies focusing on this area is suggested. PMID:22051762

  17. Detection of Mycobacterium avium subsp. paratuberculosis by a Direct In Situ PCR Method

    Science.gov (United States)

    Delgado, Fernando; Aguilar, Diana; Garbaccio, Sergio; Francinelli, Gladys; Hernández-Pando, R.; Romano, María Isabel

    2011-01-01

    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. PMID:21772965

  18. Serum BAFF levels, Methypredsinolone therapy, Epstein-Barr Virus and Mycobacterium avium subsp. paratuberculosis infection in Multiple Sclerosis patients.

    Science.gov (United States)

    Mameli, Giuseppe; Cocco, Eleonora; Frau, Jessica; Arru, Giannina; Caggiu, Elisa; Marrosu, Maria Giovanna; Sechi, Leonardo A

    2016-01-01

    Elevated B lymphocyte activating factor BAFF levels have been reported in multiple sclerosis (MS) patients; moreover, disease-modifying treatments (DMT) have shown to influence blood BAFF levels in MS patients, although the significance of these changes is still controversial. In addition, BAFF levels were reported increased during infectious diseases. In our study, we wanted to investigate on the serum BAFF concentrations correlated to the antibody response against Mycobacterium avium subspecies paratuberculosis (MAP), Epstein-Barr virus (EBV) and their human homologous epitopes in MS and in patients affected with other neurological diseases (OND), divided in Inflammatory Neurological Diseases (IND), Non Inflammatory Neurological Diseases (NIND) and Undetermined Neurological Diseases (UND), in comparison to healthy controls (HCs). Our results confirmed a statistically significant high BAFF levels in MS and IND patients in comparison to HCs but not NIND and UND patients. Interestingly, BAFF levels were inversely proportional to antibodies level against EBV and MAP peptides and the BAFF levels significantly decreased in MS patients after methylprednisolone therapy. These results implicate that lower circulating BAFF concentrations were present in MS patients with humoral response against MAP and EBV. In conclusion MS patients with no IgGs against EBV and MAP may support the hypothesis that elevated blood BAFF levels could be associated with a more stable disease. PMID:27383531

  19. Induction of matrix metalloproteinases and TLR2 and 6 in murine colon after oral exposure to Mycobacterium avium subsp. paratuberculosis.

    Science.gov (United States)

    Roderfeld, Martin; Koc, Arzu; Rath, Timo; Blöcher, Sonja; Tschuschner, Annette; Akineden, Ömer; Fischer, Marta; von Gerlach, Susanne; Goethe, Ralph; Eckelt, Elke; Meens, Jochen; Bülte, Michael; Basler, Tina; Roeb, Elke

    2012-06-01

    Mycobacterium avium subspecies paratuberculosis (MAP) is suspected to be a causative agent in Crohn's disease. Recent evidence suggests that MAP can induce the expression of Matrix Metalloproteinases (MMPs), which are the main proteases in the pathogenesis of mucosal ulcerations in inflammatory bowel disease (IBD). Within the present study, we analysed whether oral MAP exposure can induce colonic MMP expression in vivo. In MAP exposed mice MAP and spheroplasts were visualized in intramucosal leukocyte aggregates. MAP exposed mice exhibited a higher colonic expression of Mmp-2, -9, -13, -14, Timp-1, Tlr2, Tlr6, Il-1β, and Tnf-α. Cell clusters of MMP-9 positive cells adjacent to intramucosal leukocyte aggregates and CD45(+) leukocytes were identified as the major cellular sources of MMP-9. Enhanced TLR2 expression was visualized on the luminal side of colonic enterocytes. Although MAP exposure did not lead to macroscopic intestinal inflammation, the observed MAP spheroplasts in intramucosal leukocyte aggregates together with increased colonic expression of toll-like receptors, pro-inflammatory cytokines, and MMPs upon MAP exposure represents a part of the host immune response towards MAP. PMID:22289202

  20. 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. PMID:27010259

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

    DEFF Research Database (Denmark)

    Kirkeby, Carsten; Græsbøll, Kaare; Hisham Beshara Halasa, Tariq; Toft, Nils; Nielsen, Søren Saxmose

    2015-01-01

    control, in order to assist the decision-makers in planning and optimizing these programs economically. We compared six different groups of cattle (three dairy and three non-dairy) in Denmark by calculating the MES for each herd in each group.Results: The distribution of MES showed a large variation...... for MAP, given that the data used are unbiased.......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 is...

  2. Current perspectives on Mycobacterium avium subsp. paratuberculosis, Johne's disease, and Crohn's disease: a review.

    Science.gov (United States)

    Over, Ken; Crandall, Philip G; O'Bryan, Corliss A; Ricke, Steven C

    2011-05-01

    Mycobacterium avium subsp. paratuberculosis (MAP) causes the disease of cattle, Johne's. The economic impact of this disease includes early culling of infected cattle, reduced milk yield, and weight loss of cattle sold for slaughter. There is a possible link between MAP and Crohn's disease, a human inflammatory bowel disease. MAP is also a potential human food borne pathogen because it survives current pasteurization treatments. We review the current knowledge of MAP, Johne's disease and Crohn's disease and note directions for future work with this organism including rapid and economical detection, effective management plans and preventative measures. PMID:21254832

  3. 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.

  4. 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...

  5. Mycobacterium avium subsp. paratuberculosis Strains Isolated from Crohn's Disease Patients and Animal Species Exhibit Similar Polymorphic Locus Patterns

    OpenAIRE

    Ghadiali, Alifiya H.; Strother, Megan; Naser, Saleh A.; Manning, Elizabeth J. B.; Sreevatsan, Srinand

    2004-01-01

    Analysis of short sequence repeats of Mycobacterium avium subsp. paratuberculosis isolated from Crohn's disease patients identified two alleles, both of which clustered with strains derived from animals with Johne's disease. Identification of a limited number of genotypes among human strains implies the existence of human disease-associated genotypes and strain sharing with animals.

  6. Neutralization of Interleukin-10 Significantly Enhances Gamma Interferon Expression in Peripheral Blood by Stimulation with Johnin Purified Protein Derivative and by Infection with Mycobacterium avium subsp. paratuberculosis in Experimentally Infected Cattle with Paratuberculosis

    OpenAIRE

    Buza, Joram J.; Hikono, Hirokazu; Mori, Yasuyuki; Nagata, Reiko; Hirayama, Sachiyo; Bari, Abusaleh M.; Shu, Yujing; Tsuji, Noriko M.; Momotani, Eiichi

    2004-01-01

    Monoclonal antibody neutralization of interleukin-10 (IL-10) increased Johnin purified protein derivative-induced whole-blood gamma interferon (IFN-γ) secretion 23-fold and also increased IFN-γ secretion ninefold following in vitro Mycobacterium avium subsp. paratuberculosis infection of peripheral blood mononuclear cells. These results demonstrate the suppressive effect of IL-10 on immune responses to M. avium subsp. paratuberculosis infection in cattle.

  7. Relationship between Crohn's disease, infection with Mycobacterium a vium subspecies paratuberculosis and SLC11A1 gene polymorphisms in Sardinian patients

    Institute of Scientific and Technical Information of China (English)

    Leonardo A Sechi; Maria Gazouli; Lee E Sieswerda; Paola Molicotti; Niyaz Ahmed; John Ikonomopoulos; Antonio M Scanu; Daniela Paccagnini; Stefania Zanetti

    2006-01-01

    AIM: To study the association between Crohn's disease (CD),Mycobacterium avium subspecies paratuberculosis (MAP), and genetic factors by examining the role of natural resistance-associated macrophage protein 1 (NRAMP1) gene polymorphisms (now SLC11A1) in Sardinian patients with CD and controls.METHODS: Thirty-seven CD patients and 34 controls with no inflammatory bowel disease (IBD) were recruited at the University of Sassari after giving written consent. Six SCL11A1 polymorphisms previously reported to be the most significantly associated with IBD were searched.M. paratuberculosis was identified by IS900PCR and sequencing. Logistic regression was used to calculate odds ratios (OR) for the associations among CD,presence of MAP, and 6 loci described above.RESULTS: For the first time, a strong association was observed between polymorphisms at NRAMP1 locus 823C/T and CD. While CD was strongly associated with both NRAMP1 and MAP, NRAMP1 polymorphisms and MAP themselves were not correlated.CONCLUSION: Combined with previous work on the NOD2/CARD15 gene, it is clear that the interplay of genetic, infectious, and immunologic factors in the etiologyof CD is complex.

  8. Mycobacterium avium subsp. paratuberculosis Infection Causes Suppression of RANTES, Monocyte Chemoattractant Protein 1, and Tumor Necrosis Factor Alpha Expression in Peripheral Blood of Experimentally Infected Cattle

    OpenAIRE

    Buza, Joram J.; Mori, Yasuyuki; Bari, Abusaleh M.; Hikono Aodon-geril, Hirokazu; Hirayama, Sachiyo; Shu, Yujing; Momotani, Eiichi

    2003-01-01

    Blood from cattle with subclinical Mycobacterium avium subsp. paratuberculosis infection was stimulated with M. avium subsp. paratuberculosis antigens, and expression of interleukin-1β (IL-1β), tumor necrosis factor alpha (TNF-α), RANTES, monocyte chemoattractant protein 1 (MCP-1), and IL-8 was measured. Expression of TNF-α, RANTES, and MCP-1 was lower in infected than in uninfected cattle. The reduced response may weaken protective immunity and perpetuate infection.

  9. A seroepidemiological survey of Mycobacterium avium subsp. paratuberculosis in sheep from North of Portugal

    Directory of Open Access Journals (Sweden)

    Ana C Coelho

    2010-11-01

    Full Text Available The aim of this study was evaluate the risk factors for Mycobacterium avium subsp. paratuberculosis (Map seroprevalence in sheep in the North of Portugal. The effects on seroprevalence of several variables such as individual characteristics, management practices, farm characteristics, animal health, and available veterinary services were evaluated. This information was then used in a multivariable logistic regression model in order to identify risk factors for Map seropositivity. Univariable analysis was used to screen the variables used in the logistic regression model. Variables that showed p values of <0.15 were retained for the multivariable analysis. Fifteen variables were associated with paratuberculosis in univariable analysis. The multivariable logistic regression model identified a number of variables as risk factors for seropositivity like sheep pure local and/or a cross of a local breed (OR=2.02, herd size with 31-60 head (OR=2.14, culling during the Spring-Summer season (OR=1.69 and the use of an anti-parasitic treatment such as Ivermectin as the only anti-parasitic medication (OR=5.60. Potential risk factors identified in this study support current recommendations for the control of paratuberculosis.

  10. Sensitivity of mycobacterium avium subsp paratuberculosis, escherichia coli and salmonella enterica serotype typhimurium to low pH, high organic acids and ensiling

    Science.gov (United States)

    The ability of Mycobacterium avium subsp paratuberculosis (M. paratuberculosis), Salmonella enterica serotype Typhimurium (S. Typhimurium) and a commensal Escherichia coli (E. coli) isolate to persist under low pH and high organic acid conditions was determined. Die-off rates were calculated followi...

  11. 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

  12. Isolation of Mycobacterium avium subsp paratuberculosis (Map) from feral cats on a dairy farm with Map-infected cattle.

    Science.gov (United States)

    Palmer, Mitchell V; Stoffregen, William C; Carpenter, Jeremy G; Stabel, Judith R

    2005-07-01

    Paratuberculosis is an economically important disease of dairy cattle caused by Mycobacterium avium subsp. paratuberculosis (Map). The role of nonruminant, nondomestic animals in the epidemiology of paratuberculosis in cattle is unclear. To examine nonruminant, nondomestic animals for the presence of Map, 25 feral cats, nine mice (species unknown), eight rabbits (Sylvilagus floridanus), six raccoons (Procyon lotor), and three opossums (Didelphis virginiana) were collected from a mid-western dairy with known Map-infected cattle. Mycobacterium avium subsp. paratuberculosis was isolated from the mesenteric lymph node from seven of 25 (28%) feral cats. Ileum was culture-positive for three of these seven cats, and an isolation of Map was also made from the ileum of one of nine (11%) mice. Tissue samples from other species were negative as determined by Map culture; microscopic lesions consistent with paratuberculosis were not seen in any animal. Restriction fragment polymorphism analysis of isolates from cats and dairy cattle suggest interspecies transmission. The means by which interspecies transmission occurred may be through ingestion of Map-contaminated feces or waste milk or through ingestion of Map-infected prey. Shedding of Map from infected cats was not evaluated. The epidemiologic role of Map-infected feral cats on dairy farms requires further investigation. PMID:16244077

  13. Mycobacterium avium ss paratuberculosis-associated diseases: piecing the Crohn's puzzle together.

    Science.gov (United States)

    Gitlin, Laura; Borody, Thomas Julius; Chamberlin, William; Campbell, Jordana

    2012-09-01

    The relation of Mycobacterium avium ss paratuberculosis (MAP) to Crohn's Disease (CD) and other MAP-associated conditions remains controversial. New data, coupled with the analogous Helicobacter pylori (H. pylori) story, has permitted us to piece together the MAP puzzle and move forward with a more scientific way of treating inflammatory bowel disease, particularly CD. As infection moves centre stage in inflammatory bowel disease, the dated "aberrant reaction" etiology has lost scientific credibility. Now, our growing understanding of MAP-associated diseases demands review and articulation. We focus here on (1) the concept of MAP-associated diseases; (2) causality, Johne Disease, the "aberrant reaction" hypothesis; and (3) responses to published misconceptions questioning MAP as a pathogen in CD. PMID:22858515

  14. 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. PMID:24814797

  15. Expression of NRAMP1 and iNOS in Mycobacterium avium subsp. paratuberculosis naturally infected cattle.

    Science.gov (United States)

    Delgado, F; Estrada-Chávez, C; Romano, M; Paolicchi, F; Blanco-Viera, F; Capellino, F; Chavez-Gris, G; Pereira-Suárez, A L

    2010-09-01

    Paratuberculosis (PTB) is a chronic disease caused by M. avium subsp. paratuberculosis (MAP) that affects several animal species, and some studies have suggested that there may be a relationship between Crohn's disease and PTB. Significant aspects of PTB pathogenesis are not yet completely understood, such as the role of macrophages. Natural resistance-associated macrophage protein 1 (NRAMP1) and the inducible nitric oxide synthase (iNOS) molecules have shown nonspecific effects against several intracellular pathogens residing within macrophages. However, these molecules have been scarcely studied during natural infection with MAP. In this work, changes in NRAMP1 and iNOS expression were surveyed by immunohistochemistry in tissue samples from MAP-infected cattle and healthy controls. Our findings show strong specific immunolabeling against both NRAMP1 and iNOS molecules, throughout granulomatous PTB-compatible lesions in ileum and ileocaecal lymph nodes from paratuberculous cattle compared with uninfected controls, suggesting a relationship between the expression of these molecules and the pathogenesis of PTB disease. PMID:19345998

  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. Development of a novel oral vaccine against Mycobacterium avium paratuberculosis and Johne disease: A patho-biotechnological approach

    OpenAIRE

    Johnston, C.; Coffey, A.; O'Mahony, Jim; Sleator, RD

    2009-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne disease, a granulomatous enteritis of cattle and other domesticated and wild ruminant species. Johne disease is prevalent worldwide and has a significant impact on the global agricultural economy. Current vaccines against Johne are insufficient in stemming its spread, and associated side-effects prevent their widespread use in control programs. Effective and safe vaccine strategies are needed. The main purpose...

  18. Growth of Mycobacterium avium subsp. paratuberculosis, Escherichia coli, and Salmonella Enteritidis during Preparation and Storage of Yogurt

    OpenAIRE

    Cirone, K.; Huberman, Y.; Morsella, C.; L. Méndez; Jorge, M.; F Paolicchi

    2013-01-01

    The purpose of this study was to determine the viability of Mycobacterium avium subsp. paratuberculosis (MAP), Escherichia coli (E. coli), and Salmonella Enteritidis (S. Enteritidis) during preparation and refrigerated storage of yogurt. Three yogurts were prepared using pasteurized commercial milk. Each yogurt was artificially contaminated with (1) MAP, (2) E. coli + S. Enteritidis, and (3) MAP + E. coli + S. Enteritidis. Samples were taken during and after the fermentation process until day...

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

    OpenAIRE

    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 and simulating the effect of different control options at different infection prevalences. The concentration of MAP in milk was estimated using a hierarchical simulation model representing individual co...

  20. 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.

  1. A novel real-time PCR assay for specific detection and quantification of Mycobacterium avium subsp. paratuberculosis in milk with the inherent possibility of differentiation between viable and dead cells

    OpenAIRE

    Wagner Martin; Baumgartner Walter; Khol Johannes; Volgger Patrick; Dzieciol Monika; Hein Ingeborg

    2010-01-01

    Abstract Background Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of paratuberculosis (Johne's disease) in ruminants and is suggested to be one of the etiologic factors in Crohn's disease in humans. Contaminated milk might expose humans to that pathogen. The aim of the present study was to develop a novel real-time PCR assay providing the additional possibility to detect viable Mycobacterium avium subsp. paratuberculosis (MAP) based on the MAP-specific Mptb52.16 t...

  2. Detection of mycobacterium avium subsp. paratuberculosis in cheeses from small ruminants in Tuscany.

    Science.gov (United States)

    Galiero, Alessia; Fratini, Filippo; Mataragka, Antonia; Turchi, Barbara; Nuvoloni, Roberta; Ikonomopoulos, John; Cerri, Domenico

    2016-01-18

    Paratuberculosis is an infectious disease which affects mainly domestic and wild ruminants caused by Mycobacterium avium subsp. paratuberculosis (Map). Map has been associated with human diseases like Crohn disease, type-1 diabetes, sarcoidosis, multiple sclerosis and Hashimoto's thyroiditis. The aim of this study was to determine the level of Map positivity of cheeses produced in Tuscany (Italy) as an indication of human exposure to the specific pathogen. Sampling was focused on artisanal cheeses produced without commercial starter culture from raw sheep or goat milk, on small-scale farms. Samples were tested by quantitative PCR (qPCR) and culture. Map DNA was detected in 4/7 (57.14%) goat, and in 14/25 (56%) sheep cheeses by qPCR, whereas cultivation produced a positive result in only one case. This corresponded to a goat cheese that had also reacted positively by qPCR and yielded a viable Type S (sheep) strain of Map. The Map load of the tested samples based on qPCR ranged from 6×10 to 1.8×10(4)Map cells/g of cheese. The results indicate on average 56.57% and 66.6% positivity of cheese samples and farms, respectively. Hence, the type of cheeses that were analyzed within the context of this study seem to constitute a considerable source of human exposure to Map; although the question remains of whether the Map cells were present in a viable form, since positive results were almost exclusively recorded by qPCR. PMID:26555160

  3. 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

  4. Evasión molecular de la activación del macrófago bovino por Mycobacterium avium subespecie paratuberculosis

    OpenAIRE

    René Ramírez G.; Juan Maldonado E.

    2015-01-01

    RESUMENEl Mycobacterium avium subespecie paratuberculosis (MAP) es el agente causal de una enfermedad granulomatosica crónica, que afecta el tracto gastrointestinal de rumiantes domesticos y salvajes, conocida como la enfermedad de Johne o paratuberculosis. MAP es un microorganismo de crecimiento lento en cultivo, no obstante sobrevive in vivo en células fagocíticas mononucleares de los rumiantes, bajo condiciones de susceptibilidad individual, virulencia de la cepa infectante y estado inmune...

  5. 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.

  6. Deteksi Mycobacterium Avium Subspesies Paratuberculosis pada Susu Pasturisasi yang Dijual di Bogor

    Directory of Open Access Journals (Sweden)

    Widagdo Sri Nugroho

    2010-06-01

    Full Text Available Mycobacterium avium subspesies paratuberculosis (MAP is a thermal tolerant bacteria. The presenceof these bacteria in pasteurized dairy milk is associated with infectious bowel disease in human known asCrohn’s disease. The aim of this study was to detect MAP in pasteurized dairy milk sold in Bogor. Fourtytwo samples of plain flavoured milk (180–250 ml from 7 producers were bought from supermarkets inBogor. The presence of MAP was detected by isolation and conventional polymerase chain reaction (PCRusing IS 900 and F57. Bacterial isolation were done by Herrold’s egg yolk medium with mycobactine J(HEYMj and without mycobactin J (HEYM and incubated at 37°C for 20 weeks. The DNA extraction ofall pasteurized dairy milk samples were conducted by DNeasy® Tissue Kit. Amplification conditionsfor PCR were: 1 cycle at 94°C for 10 minutes, 40 cycles at 94°C for 1 minute, 58°C for 1 minute, and72°C for 3 minutes, and 1 cycle at 72°C for 7 minutes. After 20 weeks of incubation, there were no sign ofMAP which grew in all isolation mediums. The PCR IS 900 and F57 did not detect the DNA band of thetarget. In the conclusion, there was no MAP detected in pasteurized dairy milk sold in Bogor.

  7. On deaf ears, Mycobacterium avium paratuberculosis in pathogenesis Crohn’s and other diseases

    Science.gov (United States)

    Davis, William C

    2015-01-01

    The historic suggestion that Mycobacterium avium subsp. paratuberculosis (Map) might be a zoonotic pathogen was based on the apparent similarity of lesions in the intestine of patients with Crohn’s disease (CD) with those present in cattle infected with Map, the etiological agent of Johne’s disease. Reluctance to fully explore this possibility has been attributed to the difficulty in demonstrating the presence of Map in tissues from patients with CD. Advances in technology have resolved this problem and revealed the presence of Map in a significant proportion of patients with CD and other diseases. The seminal finding from recent investigations, however, is the detection of Map in healthy individuals with no clinical signs of disease. The latter observation indicates all humans are susceptible to infection with Map and lends support to the thesis that Map is zoonotic, with a latent stage of infection similar to tuberculosis, where infection leads to the development of an immune response that controls but does not eliminate the pathogen. This clarifies one of the reasons why it has been so difficult to document that Map is zoonotic and associated with the pathogenesis of CD and other diseases. As discussed in the present review, a better understanding of the immune response to Map is needed to determine how infection is usually kept under immune control during the latent stage of infection and elucidate the triggering events that lead to disease progression in the natural host and pathogenesis of CD and immune related diseases in humans. PMID:26730151

  8. Application of multiple laboratory tests for Mycobacterium avium ssp. paratuberculosis detection in Crohn's disease patient specimens.

    Science.gov (United States)

    Banche, Giuliana; Allizond, Valeria; Sostegni, Raffaello; Lavagna, Alessandro; Bergallo, Massimiliano; Sidoti, Francesca; Daperno, Marco; Rocca, Rodolfo; Cuffini, Anna Maria

    2015-07-01

    The difficulties involved in detecting and enumerating Mycobacterium avium subsp. paratuberculosis (MAP) as a pathogen potentially involved in Crohn's disease (CD) are well known. This study aimed to improve this situation through the application of multiple laboratory diagnostic tests to detect and isolate this bacterium from different specimens collected from CD-patients and non-CD subjects as controls. A total of 120 samples (terminal ileum and colon biopsies, blood and stool) were obtained from 19 CD-patients and from 11 individuals who did not have a clinicopathological diagnosis of CD (non-CD controls) attending for ileocolonoscopy. All samples were processed by staining techniques, culture on both solid and liquid media, and Insertion Sequence 900/F57 real-time PCR. The MAP frequency in CD-patients was found in a significantly greater proportion than in non-CD subjects; the most positive samples were biopsies from CD-patients tested by real-time PCR. MAP detection in biopsies, and in the other samples, by applying multiple and validated laboratory diagnostic tests, could be a marker of active infection, supporting MAP involvement in CD. PMID:26147146

  9. Crohn's disease and Mycobacterium avium subsp. paratuberculosis: the need for a study is long overdue.

    Science.gov (United States)

    Davis, William C; Madsen-Bouterse, Sally A

    2012-01-15

    The initial suggestion that Mycobacterium avium subsp. paratuberculosis (Map) might be involved in the pathogenesis of Crohn's disease (CD) was based on the apparent similarity of lesions in the intestine of patients with CD with those present in cattle infected with Map, the etiological agent of Johne's disease (JD). Recent investigations have now revealed the presence of Map or Map DNA in blood or lesions from adults and children with CD. Of special interest, Map has also been found in patients with other diseases as well as healthy subjects. The latter observations indicate all humans are susceptible to infection with Map and that, like with other mycobacterial pathogens such as Mycobacterium tuberculosis, infection does not invariably lead to development of clinical disease but rather development of a persistent latent stage of infection where an immune response controls but does not eliminate the pathogen. Limited information has been obtained on the immune response to Map in healthy subjects and patients with CD. Understanding how Map may be involved in the pathogenesis of CD will require a better understanding of the immune response to Map in one of its common hosts as well as healthy humans and patients with CD. PMID:22209202

  10. 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. PMID:19681906

  11. Genome-Wide Diversity and Phylogeography of Mycobacterium avium subsp. paratuberculosis in Canadian Dairy Cattle.

    Science.gov (United States)

    Ahlstrom, Christina; Barkema, Herman W; Stevenson, Karen; Zadoks, Ruth N; Biek, Roman; Kao, Rowland; Trewby, Hannah; Haupstein, Deb; Kelton, David F; Fecteau, Gilles; Labrecque, Olivia; Keefe, Greg P; McKenna, Shawn L B; Tahlan, Kapil; De Buck, Jeroen

    2016-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative bacterium of Johne's disease (JD) in ruminants. The control of JD in the dairy industry is challenging, but can be improved with a better understanding of the diversity and distribution of MAP subtypes. Previously established molecular typing techniques used to differentiate MAP have not been sufficiently discriminatory and/or reliable to accurately assess the population structure. In this study, the genetic diversity of 182 MAP isolates representing all Canadian provinces was compared to the known global diversity, using single nucleotide polymorphisms identified through whole genome sequencing. MAP isolates from Canada represented a subset of the known global diversity, as there were global isolates intermingled with Canadian isolates, as well as multiple global subtypes that were not found in Canada. One Type III and six "Bison type" isolates were found in Canada as well as one Type II subtype that represented 86% of all Canadian isolates. Rarefaction estimated larger subtype richness in Québec than in other Canadian provinces using a strict definition of MAP subtypes and lower subtype richness in the Atlantic region using a relaxed definition. Significant phylogeographic clustering was observed at the inter-provincial but not at the intra-provincial level, although most major clades were found in all provinces. The large number of shared subtypes among provinces suggests that cattle movement is a major driver of MAP transmission at the herd level, which is further supported by the lack of spatial clustering on an intra-provincial scale. PMID:26871723

  12. Mycobacterium avium subsp. paratuberculosis detection in individual and bulk tank milk samples from bovine herds and caprine flocks.

    Science.gov (United States)

    Favila-Humara, Lucía C; Chávez-Gris, Gilberto G; Carrillo-Casas, Erika M; Hernández-Castro, Rigoberto

    2010-04-01

    Paratuberculosis, or Johne's disease, is caused by Mycobacterium avium subsp. paratuberculosis (Map), and it generates great economic losses for the dairy industry worldwide. In humans, Map has been associated with Crohn's disease. Mexico has unknown paratuberculosis prevalence, and yet, control programs have not been applied. This study aimed to determine the presence of Map in milk samples from seropositive goats and cows and bulk tank milk samples from herds previously designated Map-infected using indirect enzyme-linked immunosorbent assay. Map DNA was detected in 100% of the bulk tank milk samples of 14 bovine herds and 3 caprine flocks using a modified insertion sequence 900 polymerase chain reaction (PCR). Additionally, Map DNA was detected in 100% of the individual milk samples from 10 cows and 8 goats. Further, based on the findings of the experimental insertion sequence 900 PCR assessment, evaluation of bulk tank and individual milk samples through a type-specific PCR was performed, which confirmed our previous findings and revealed that 56.25% cow and 63.63% goat milk had concurrent infections of the C, I, and S types. Out of 14 bulk tank milk samples, 10 had viable mycobacteria. Paratuberculosis was detected at a high frequency in cow and goat milk, which suggests that raw milk ingestion represents a potential risk of Map infection. PMID:19911881

  13. Longitudinal study of interferon-gamma, serum antibody and milk antibody responses in cattle infected with Mycobacterium avium subsp paratuberculosis

    DEFF Research Database (Denmark)

    Huda, A.; Jungersen, Gregers; Lind, Peter

    During a 2-year study period, 252 animals from dairy herds infected with Mycobacterium avium subsp. paratuberculosis, and 119 animals from non-infected herds were subjected to repeated blood and faecal sampling. Animals were retrospectively grouped by infection status as infected, exposed (culture......-blood lymphocytes (IFN-gamma test), and measurement of antibody responses against M. paratuberculosis in serum and milk by an in-house absorbed ELISA. The IFN-gamma test diagnosed higher proportions of infected and exposed animals than the antibody ELISAs. The highest sensitivity of IFN-gamma test was in infected...... compared with repeated samplings showed better performance of the IFN-gamma test by repeated samplings, and the milk antibody ELISA in animals of 3+ years of age performed significantly better with repeated sampling compared with single sampling. In conclusion, the IFN-gamma test may be applied for...

  14. 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...... depends on the prevalence among contributing herds and individuals. Considerable variation of MAP in bulk tank milk (BTM) and individual cow’s milk (IM) is reported, but factors associated with MAP occurrence in milk at farm level have not been described. This study systematically reviewed published...

  15. Novel Monoclonal Antibody and Peptide Binders for Mycobacterium avium subsp. paratuberculosis and Their Application for Magnetic Separation

    OpenAIRE

    Lorna M O'Brien; Stewart, Linda D.; Strain, Sam A. J.; Grant, Irene R

    2016-01-01

    The generation of novel Mycobacterium avium subsp. paratuberculosis (MAP)-specific monoclonal antibodies and phage-display derived peptide binders, along with their application for the magnetic separation (MS) of MAP cells, is described. Our aim was to achieve even greater MAP capture capability than is possible with peptide-mediated magnetic separation (PMS) using a 50:50 mix of biotinylated-aMp3 and biotinylated-aMptD peptide-coated beads. Gamma-irradiated whole MAP cells and ethanol extrac...

  16. Isolation of High-Affinity Single-Chain Antibodies against Mycobacterium avium subsp. paratuberculosis Surface Proteins from Sheep with Johne's Disease

    Science.gov (United States)

    Johne’s disease, due to infection with Mycobacterium avium subsp. paratuberculosis, causes significant economic losses to the livestock farming industry. Improved investigative and diagnostic tools - necessary to understand disease processes and to identify sub-clinical infection - are much sought a...

  17. 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; Tondo, Annalucia; Nielsen, Søren Saxmose

    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...

  18. Chemical decontamination with n-acetyl-l-cysteine-sodium hydroxide improves recovery of viable Mycobacterium avium subsp. paratuberculosis organisms from cultured milk

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is shed into milk and feces of cows with advanced Johne’s disease, allowing transmission of MAP among animals. The objective of this study was to formulate an optimized protocol for the isolation of MAP from milk. Parameters investigated included che...

  19. 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.

  20. A Mycobacterium avium subsp. paratuberculosis Predicted Serine Protease Is Associated with Acid Stress and Intraphagosomal Survival.

    Science.gov (United States)

    Kugadas, Abirami; Lamont, Elise A; Bannantine, John P; Shoyama, Fernanda M; Brenner, Evan; Janagama, Harish K; Sreevatsan, Srinand

    2016-01-01

    The 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 macrophages and MAC-T cells that 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 mc(2) 155, was designed to express MAP0403 and when exposed to either macrophages or in vitro acid stress had increased bacterial cell viability, which corresponds to maintenance of intra-bacterial pH in acidic (pH = 5) conditions, compared to the parent strain. 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. PMID:27597934

  1. Short communication: Evaluation of sampling socks for detection of Mycobacterium avium ssp. paratuberculosis on dairy farms.

    Science.gov (United States)

    Wolf, R; Orsel, K; De Buck, J; Kanevets, U; Barkema, H W

    2016-04-01

    Mycobacterium avium ssp. paratuberculosis (MAP) causes Johne's disease, a production-limiting disease in cattle. Detection of infected herds is often done using environmental samples (ES) of manure, which are collected in cattle pens and manure storage areas. Disadvantages of the method are that sample accuracy is affected by cattle housing and type of manure storage area. Furthermore, some sampling locations (e.g., manure lagoons) are frequently not readily accessible. However, sampling socks (SO), as used for Salmonella spp. testing in chicken flocks, might be an easy to use and accurate alternative to ES. The objective of the study was to assess accuracy of SO for detection of MAP in dairy herds. At each of 102 participating herds, 6 ES and 2 SO were collected. In total, 45 herds had only negative samples in both methods and 29 herds had ≥1 positive ES and ≥1 positive SO. Furthermore, 27 herds with ≥1 positive ES had no positive SO, and 1 herd with no positive ES had 1 positive SO. Bayesian simulation with informative priors on sensitivity of ES and MAP herd prevalence provided a posterior sensitivity for SO of 43.5% (95% probability interval=33-58), and 78.5% (95% probability interval=62-93) for ES. Although SO were easy to use, accuracy was lower than for ES. Therefore, with improvements in the sampling protocol (e.g., more SO per farm and more frequent herd visits), as well as improvements in the laboratory protocol, perhaps SO would be a useful alternative for ES. PMID:26851860

  2. 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.

  3. 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

  4. Rapid and Sensitive Detection of Mycobacterium avium subsp. paratuberculosis in Bovine Milk and Feces by a Combination of Immunomagnetic Bead Separation-Conventional PCR and Real-Time PCR

    OpenAIRE

    Khare, Sangeeta; Thomas A Ficht; Renato L. Santos; Romano, Juan; Ficht, Allison R.; Zhang, Shuping; Grant, Irene R.; Libal, Melissa; Hunter, David; Adams, L. Garry

    2004-01-01

    Immunomagnetic bead separation coupled with bead beating and real-time PCR was found to be a very effective procedure for the isolation, separation, and detection of Mycobacterium avium subsp. paratuberculosis from milk and/or fecal samples from cattle and American bison. Samples were spiked with M. avium subsp. paratuberculosis organisms, which bound to immunomagnetic beads and were subsequently lysed by bead beating; then protein and cellular contaminants were removed by phenol-chloroform-i...

  5. Avian mycobacteriosis caused by Mycobacterium avium subspecies avium in four ornamental birds and in vitro drug sensitivity testing of isolates.

    Science.gov (United States)

    Stepień-Pyśniak, Dagmara; Puk, Krzysztof; Guz, Leszek; Wawrzyniak, Agata; Marek, Agnieszka; Kosikowska, Urszula

    2016-01-01

    Avian tuberculosis, one of the most important diseases affecting various species of birds, is most often caused by Mycobacterium (M.) avium. This report describes cases of M. avium subsp. avium (MAA) infection in a white-crested Holland dwarf rooster, a male and a female golden pheasant and a male peacock. We also investigated the prevalence of mycobacteria in 60 other birds and 40 alpacas. Tissue samples of necropsied birds were cultured for mycobacteria. From non-necropsied 60 other birds and alpacas only faecal samples were collected. Clinical signs in the affected white-crested Holland cock included gradual loss of body weight and hoarse attempts at crowing during its last 3 weeks, with a dramatic loss of body condition and depression over the final week. Only slight weakening was observed in the peacock just before its death, and the golden pheasants died suddenly. Diagnosis was confirmed by microbiological, molecular and pathological results. Mycobacterium avium subsp. avium strains were isolated from the internal organs of the affected birds. Only one faecal sample from 60 other birds was culture- and PCR-positive for M. avium subsp. avium, while another one was only PCR-positive for M. chelonae. We did not isolate any Mycobacterium spp. from faecal samples of alpacas and all of them were PCR-negative. All 18 isolated M. avium strains were resistant to rifampicin, isoniazid, ethambutol, ethionamide, capreomycin and ofloxacin, and susceptible to cycloserine and streptomycin. PMID:26904899

  6. ZAP-70, CTLA-4 and proximal T cell receptor signaling in cows infected with Mycobacterium avium subsp. paratuberculosis.

    Science.gov (United States)

    Leite, Fernando L; Eslabão, Livia B; Pesch, Bruce; Bannantine, John P; Reinhardt, Timothy A; Stabel, Judith R

    2015-09-15

    Paratuberculosis is a chronic intestinal disease of ruminant animals caused by Mycobacterium avium subsp. paratuberculosis (MAP). A hallmark of paratuberculosis is a transition from a cell-mediated Th1 type response to a humoral Th2 response with the progression of disease from a subclinical to clinical state. The objective of this study was to investigate the expression of two crucial molecules in T cell function, ZAP-70 (zeta-chain-associated protein of 70 kDa) and CTLA-4 (cytotoxic T-lymphocyte antigen-4), in cows naturally infected with MAP. Peripheral blood mononuclear cells (PBMCs) isolated from control non-infected cows (n=5), and cows in subclinical (n=6) and clinical stages of paratuberculosis (n=6) were cultured alone (medium only), and with concanavalin A, and a whole cell sonicate of MAP for 24, 72 and 144 h to measure the dynamic changes of ZAP-70 and CTLA-4 expression on CD4, CD8, and gamma delta (γδ) T cells. Flow cytometry was also performed to measure ZAP-70 phosphorylation to examine proximal T cell receptor signaling in animals of different disease status. The surface expression of CTLA-4 was increased in animals in subclinical stage of infection while levels of ZAP-70 were decreased in CD4+ T cells of both subclinical and clinical animals, indicating a change in T cell phenotype with disease state. Interestingly, proximal T cell receptor signaling was not altered in infected animals. This study demonstrated changes in crucial signaling molecules in animals infected with MAP, thereby elucidating T cell alterations associated with disease progression. PMID:26163934

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

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    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.

  8. Detection and Confirmation of Mycobacterium avium subsp. paratuberculosis in Direct Quantitative PCR Positive Fecal Samples by the Manual Fluorescent MGIT Culture System

    OpenAIRE

    KAWAJI, Satoko; Nagata, Reiko; Mori, Yasuyuki

    2013-01-01

    ABSTRACT An efficient protocol for the manual fluorescent MGIT culture system combined with rapid confirmation of Mycobacterium avium subsp. paratuberculosis (MAP) growth in the broth culture was established and evaluated for the detection of viable MAP in direct quantitative PCR (QPCR) positive bovine feces. Manually detected fluorescence emissions from MGIT tubes were analyzed objectively using an open source software, ImageJ. For molecular confirmation of MAP growth, DNA samples harvested ...

  9. Paratuberculosis: decrease in milk production of German Holstein dairy cows shedding Mycobacterium avium ssp. paratuberculosis depends on within-herd prevalence.

    Science.gov (United States)

    Donat, K; Soschinka, A; Erhardt, G; Brandt, H R

    2014-05-01

    Paratuberculosis impairs productivity of infected dairy cows because of reduced milk production and fertility and enhanced risk of culling. The magnitude of the milk yield depression in individual cows is influenced by factors such as parity, the stage of the disease and the choice of test used. The objectives of this case-control study were to substantiate the influence of the different levels of the within-herd prevalence (WHP) on individual milk yield of fecal culture (FC)-positive cows (FC+) compared with FC-negative herd-mates (FC-), and to estimate the magnitude of the deviation of the milk yield, milk components and somatic cell count (SCC) in an FC-based study. Of a total of 31 420 cows from 26 Thuringian dairy herds tested for paratuberculosis by FC, a subset of 1382 FC+ and 3245 FC- with milk recording data were selected as cases and controls, respectively. The FC- cows were matched for the same number and stage of lactation (±10 days in milk) as one FC+ from the same herd. Within a mixed model analysis using the fixed effects of Mycobacterium avium ssp. paratuberculosis (MAP) status, lactation number, days in milk, prevalence class of farm and the random effect of farm on milk yield per day (kg), the amount of fat and protein (mg/dl) and lactose (mg/dl) as well as the SCC (1000/ml) were measured. On the basis of least square means, FC+ cows had a lower test-day milk yield (27.7±0.6 kg) compared with FC- (29.0±0.6 kg), as well as a lower milk protein content and a slightly diminished lactose concentration. FC status was not associated with milk fat percentage or milk SCC. In FC+ cows, reduction in milk yield increased with increasing WHP. An interaction of FC status and farm was found for the test-day milk yield, and milk protein percentage, respectively. We conclude that the reduction in milk yield of FC+ cows compared with FC- herd-mates is significantly influenced by farm effects and depends on WHP class. Owners of MAP-positive dairy herds may

  10. Detection of Mycobacterium avium subspecies paratuberculosis in surgicalpathology blocks from patients with Crohn's disease in China

    Institute of Scientific and Technical Information of China (English)

    Jun Cheng; Tim Bull; Nazira Sumar; Joe Sheridan; Peter Dalton; John Hermon-Taylor

    2000-01-01

    AIM To determine whether MAP can be detected in archival paraffin embedded full thickness samples ofintestinal tissue from patients in China with Crohn's disease (CD), ulcerative colitis (UC), and in controlsubjects (NIBD) having surgery for bowel cancer.METHODS Optimized procedures for the removal of paraffin, recovery of tissue and access to MAP DNA,followed by MAP-specific nested IS900 PCR. Confirmation of specific amplification by Southern blotting andDNA sequencing.RESULTS IS900 PCR positive tests identified MAP in 9 (69%) of 13 CD, 1 of 3 UC and 2 (14%) of 14NIBD in the presence of correctly reporting positive and negative sample and reagent control reactions. DNAsequence analysis of the 298bp IS900 PCR amplification product from MAP in 2 Chinese CD patientsdemonstrated 99% homology with the GenBank IS900 sequence accession number X16293.CONCLUSION Although larger numbers of Chinese samples need to be studied, these initial results areconsistent with an exposure of human populations in China to MAP, and an involvement of this pathogen inchronic inflammation of the intestine of the Crohn's disease type. The results are in agreement with similarpositive studies reported from China, from Western Europe and elsewhere.

  11. Effects of the probiotic Lactobacillus animalis in murine Mycobacterium avium subspecies paratuberculosis infection

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    Karunasena Enusha

    2013-01-01

    Full Text Available Abstract Background MAP is a suspected zoonotic pathogen and the causative agent of Johne’s Disease in cattle and other ruminant animals. With over $1 billion dollars in loss to the dairy industry due to Johne’s Disease, efforts to eliminate or reduce MAP from cattle are of importance. The purpose of this study was to determine if daily intake of probiotics could eliminate or reduce Johne’s Disease associated symptoms and pathogenesis by MAP. Post infection, animals are often asymptomatic carriers with limited shedding of the pathogen, proving early detection to be difficult. Disease and symptoms often appear 3–4 years after infection with antibiotic treatment proving ineffective. Symptoms include chronic gastrointestinal inflammation leading to severe weight-loss from poor feed and water intake cause a wasting disease. These symptoms are similar to those found in individuals with Crohn’s Disease (CD; MAP has been implicated by not proven to be the causative agent of CD. Probiotics administered to livestock animals, including dairy and beef cattle have demonstrated improvements in cattle performance and health. Our objectives included determining the benefits of Lactobacillus animalis (strain name: NP-51 in MAP infected BALB/c mice by evaluating systemic and gastrointestinal response by the host and gut microbiota. Male and female animals were fed 1×106 CFU/g probiotics in sterile, powdered mouse chow daily and infected with 1 × 107 CFU/ml MAP and compared to controls. Animals were evaluated for 180 days to assess acute and chronic stages of disease, with sample collection from animals every 45 days. MAP concentrations from liver and intestinal tissues were examined using real time-PCR methods and the expression of key inflammatory markers were measured during MAP infection (interferon-gamma [IFN-Υ], Interleukin-1α, IL-12, IL-10, IL-6, and Tumor necrosis factor alpha [TNF-α]. Results Our results demonstrate administration of probiotics reduces production of IFN-Υ and IL-6 while increasing TNF-α and IL-17 in chronic disease; healthful immune responses that reduce chronic inflammation associated to MAP infection. Conclusions We observed that the immune system’s response in the presence of probiotics to MAP contributes towards host health by influencing the activity of the immune system and gut microbial populations.

  12. Rapid and sensitive detection of Mycobacterium avium subsp. paratuberculosis in bovine milk and feces by a combination of immunomagnetic bead separation-conventional PCR and real-time PCR.

    Science.gov (United States)

    Khare, Sangeeta; Ficht, Thomas A; Santos, Renato L; Romano, Juan; Ficht, Allison R; Zhang, Shuping; Grant, Irene R; Libal, Melissa; Hunter, David; Adams, L Garry

    2004-03-01

    Immunomagnetic bead separation coupled with bead beating and real-time PCR was found to be a very effective procedure for the isolation, separation, and detection of Mycobacterium avium subsp. paratuberculosis from milk and/or fecal samples from cattle and American bison. Samples were spiked with M. avium subsp. paratuberculosis organisms, which bound to immunomagnetic beads and were subsequently lysed by bead beating; then protein and cellular contaminants were removed by phenol-chloroform-isopropanol extraction prior to DNA precipitation. DNA purified by this sequence of procedures was then analyzed by conventional and real-time IS900-based PCR in order to detect M. avium subsp. paratuberculosis in feces and milk. By use of this simple and rapid technique, 10 or fewer M. avium subsp. paratuberculosis organisms were consistently detected in milk (2-ml) and fecal (200-mg) samples, making this sensitive procedure very useful and cost-effective for the diagnosis of clinical and subclinical Johne's disease (paratuberculosis) compared to bacteriological culture, which is constrained by time, labor, and expense under diagnostic laboratory conditions. PMID:15004056

  13. Rational Discovery of T Helper Epitopes Specific for Bovine Infections with Mycobacterium avium ssp paratuberculosis

    DEFF Research Database (Denmark)

    Lundegaard, Claus; Aagaard, C.; Ussery, David;

    2011-01-01

    Paratuberculosis, a chronic wasting disease in ruminants, is causing significant losses to both EU dairy and meat producers, due to a decreasing milk yield, loss of body weight and early replacements. The absence of adequate diagnostic tools for early detection of subclinically infected livestock...... goal is to combine positive responding peptides with new promising adjuvants in order to develop effective bovine MAP vaccines which do not cross react with traditional skin tests for M. bovis infections....

  14. Overt Mycobacterium avium subsp. paratuberculosis Infection: An Infrequent Occurrence in Archived Tissue from False TB Reactor Cattle in Michigan, USA

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    Scott D. Fitzgerald

    2011-01-01

    Full Text Available The objective of this study was to retrospectively determine whether or not cattle from the state of Michigan which were classified as bovine tuberculosis reactors, based on currently approved field and laboratory testing methods, were overtly infected with Mycobacterium avium subsp. paratuberculosis (MAP. Included in this study were 384 adult cattle submitted to the Diagnostic Center for Population and Animal Health over a seven-year period. Cattle were tested utilizing standard methods to confirm that all cattle were lesion and culture negative for infection with Mycobacterium bovis at postmortem examination. Retrospective analysis of formalin-fixed, paraffin-embedded sections of ileum and ileocecal lymph node were evaluated by histopathology, acid-fast staining, and PCR assays to detect MAP. Overall, only 1.04 percent of cattle showed overt infection with MAP on visual examination of sections of ileum and/or ileo-cecal lymph node. This increased slightly to 2.1 percent of cattle likely infected with MAP after additional testing using a PCR assay. Based on these results, we found no evidence that overt infection with MAP plays a major role in the false tuberculosis reactor test results for cattle examined in this study.

  15. Mycobacterium avium subsp paratuberculosis--incidences in milk and milk products, their isolation, enumeration, characterization, and role in human health.

    Science.gov (United States)

    Patel, Ami; Shah, Nihir

    2011-12-01

    Mycobacterium avium subsp paratuberculosis (MAP), excreted in the feces and milk, is reported to be not easily inactivated by pasteurization and thermal treatments as other bacteria infecting humans and animals do. The D values of all MAP strains tested were considerably higher than those published for other pathogens. Culturing techniques for this organism are labor intensive. Although an increasing amount of scientific evidence suggests that this organism can be responsible for at least some cases of Crohn's disease (CD), there is controversy about MAP being a cause of CD in humans. In general, although some studies have described an association between the presence of MAP and CD, the role of Mycobacterium species and MAP in the etiology of this human disease remains unestablished. Although published reports indicate that it may not be completely inactivated by pasteurization of milk, the effectiveness of increasing the time or temperature in the pasteurization process has not been established and hence any potential benefit to human health cannot be determined. This article summarizes the incidences of MAP in milk and milk products with respect to human health and brief discussion of various serological as well as molecular techniques used for their isolation, enumeration, and characterization. PMID:21620785

  16. Longitudinal evaluation of diagnostics in experimentally infected young calves during subclinical and clinical paratuberculosis.

    Science.gov (United States)

    Mortier, Rienske A R; Barkema, Herman W; Orsel, Karin; Muench, Gregory P; Bystrom, Janet M; Illanes, Oscar; De Buck, Jeroen

    2015-12-01

    Five calves were inoculated orally at 2 weeks of age with a dose of 5 × 10(9) colony-forming units of Mycobacterium avium subspecies paratuberculosis (MAP) on 2 consecutive days. Two calves developed clinical Johne's disease at 12 and 16 months of age after being consistently positive for MAP on fecal culture and antibody enzyme-linked immunosorbent assay (ELISA), starting 2 to 3 weeks and 4 to 5 months after inoculation, respectively. PMID:26663923

  17. Development of improved method for isolation of Mycobacterium avium subsp. paratuberculosis from bulk tank milk: Effect of age of milk, centrifugation, and decontamination

    OpenAIRE

    Gao, Anli; Odumeru, Joseph; Raymond, Melinda; Mutharia, Lucy

    2005-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease in cattle and it has been suggested that this organism may be associated with Crohn’s disease in humans. Cows at the advanced stage of the disease shed this organism into both their milk and feces. The objective of this study was to develop a more efficient procedure for isolating MAP from bulk tank raw milk. Bulk tank raw milk (50 mL) samples 3 to 13 d old after collection without spiking were investi...

  18. The modification and evaluation of an ELISA test for the surveillance of Mycobacterium avium subsp. paratuberculosis infection in wild ruminants

    Directory of Open Access Journals (Sweden)

    Pruvot Mathieu

    2013-01-01

    Full Text Available Abstract Background Enzyme-linked immunosorbent assay (ELISA is often used to test wildlife samples for Mycobacterium avium subsp. paratuberculosis (MAP infection. However, commercially available kits are only validated for use with domestic ruminant species. A literature review was performed to document the current use of MAP serum ELISA in wild and semi-domestic ruminants. We then modified and evaluated a commercial ELISA kit (IDEXX Mycobacterium paratuberculosis Antibody Test Kit for use with species for which it was not originally developed: elk (Cervus elaphus, bison (Bison bison and caribou (Rangifer tarandus. We tested the affinity of different conjugates for immunoglobulin G (IgG isolated from these species, performed checkerboard tests to determine the optimal dilutions of samples and conjugates, and established cut-off values using two different methods: a Receiver Operational Curve on a panel of known samples for elk, and an alternate method involving a panel of unknown serum samples for the three species. Results We found that the anti-bovine conjugate included in the IDEXX ELISA kit has limited affinity for elk, bison, and caribou IgG. Protein G showed good affinity for IgG of all three species, while anti-deer conjugate also bound elk and caribou IgG. Using Protein G with elk serum, a cut-off sample-to-positive (S/P value of 0.22 was selected, resulting in a sensitivity and specificity of 73% and 90%, respectively, whereas, using an anti-deer conjugate with elk serum, an S/P cut-off value of 0.29 gave a sensitivity of 68%, with 100% specificity. Cut-off values for bison and caribou using the Protein G conjugate were 0.17 and 0.25 respectively. Conclusions Due to incomplete reporting and a lack of test validation, it is difficult to critically appraise results of many sero-surveys that have previously been done for MAP in wildlife. Commercial ELISA kits may have limited or no capacity to detect antibodies from species other than for

  19. Rational Discovery of T Helper Epitopes Specific for Bovine Infections with Mycobacterium avium ssp paratuberculosis

    DEFF Research Database (Denmark)

    Lundegaard, Claus; Aagaard, C.; Ussery, David; Andersen, P. L.; Jungersen, Gregers

    2011-01-01

    Paratuberculosis, a chronic wasting disease in ruminants, is causing significant losses to both EU dairy and meat producers, due to a decreasing milk yield, loss of body weight and early replacements. The absence of adequate diagnostic tools for early detection of subclinically infected livestock...... bioinformatics tools has been used to identify peptides in the MAP genome, which are predicted to bind to bovine MHC (BoLA) class II antigen presenting molecules. Comparative genomics tools has been used to further select MAP specific peptides 100% conserved in the two MAP strains and with low similarity to...... goal is to combine positive responding peptides with new promising adjuvants in order to develop effective bovine MAP vaccines which do not cross react with traditional skin tests for M. bovis infections....

  20. Polymorphisms in the gene encoding bovine interleukin-10 receptor alpha are associated with Mycobacterium avium ssp. paratuberculosis infection status

    Directory of Open Access Journals (Sweden)

    Kelton David F

    2010-04-01

    Full Text Available Abstract Background Johne's disease is a chronic inflammatory bowel disease (IBD of ruminants caused by Mycobacterium avium ssp. paratuberculosis (MAP. Since this pathogen has been implicated in the pathogenesis of human IBDs, the goal of this study was to assess whether single nucleotide polymorphism (SNPs in several well-known candidate genes for human IBD are associated with susceptibility to MAP infection in dairy cattle. Methods The bovine candidate genes, interleukin-10 (IL10, IL10 receptor alpha/beta (IL10RA/B, transforming growth factor beta 1 (TGFB1, TGFB receptor class I/II (TGFBR1/2, and natural resistance-associated macrophage protein 1 (SLC11A1 were sequenced for SNP discovery using pooled DNA samples, and the identified SNPs were genotyped in a case-control association study comprised of 242 MAP negative and 204 MAP positive Holstein dairy cattle. Logistic regression was used to determine the association of SNPs and reconstructed haplotypes with MAP infection status. Results A total of 13 SNPs were identified. Four SNPs in IL10RA (984G > A, 1098C > T, 1269T > C, and 1302A > G were tightly linked, and showed a strong additive and dominance relationship with MAP infection status. Haplotypes AGC and AAT, containing the SNPs IL10RA 633C > A, 984G > A and 1185C > T, were associated with an elevated and reduced likelihood of positive diagnosis by serum ELISA, respectively. Conclusions SNPs in IL10RA are associated with MAP infection status in dairy cattle. The functional significance of these SNPs warrants further investigation.

  1. Effects of biological and methodological factors on volatile organic compound patterns during cultural growth of Mycobacterium avium ssp. paratuberculosis.

    Science.gov (United States)

    Küntzel, Anne; Fischer, Sina; Bergmann, Andreas; Oertel, Peter; Steffens, Markus; Trefz, Phillip; Miekisch, Wolfram; Schubert, Jochen K; Reinhold, Petra; Köhler, Heike

    2016-01-01

    Mycobacterium avium ssp. paratuberculosis (MAP) causes chronic granulomatous enteritis in ruminants. Bacterial growth is still the diagnostic 'gold standard', but is very time consuming. MAP-specific volatile organic compounds (VOCs) above media could accelerate cultural diagnosis. The aim of this project was to assess the kinetics of a VOC profile linked to the growth of MAP in vitro. The following sources of variability were taken into account: five different culture media, three different MAP strains, inoculation with different bacterial counts, and different periods of incubation. Needle-trap microextraction was employed for pre-concentration of VOCs, and gas chromatography-mass spectrometry for subsequent analysis. All volatiles were identified and calibrated by analysing pure references at different concentration levels. More than 100 VOCs were measured in headspaces above MAP-inoculated and control slants. Results confirmed different VOC profiles above different culture media. Emissions could be assigned to either egg-containing media or synthetic ingredients. 43 VOCs were identified as potential biomarkers of MAP growth on Herrold's Egg Yolk Medium without significant differences between the tree MAP strains. Substances belonged to the classes of alcohols, aldehydes, esters, ketones, aliphatic and aromatic hydrocarbons. With increasing bacterial density the VOC concentrations above MAP expressed different patterns: the majority of substances increased (although a few decreased after reaching a peak), but nine VOCs clearly decreased. Data support the hypotheses that (i) bacteria emit different metabolites on different culture media; (ii) different MAP strains show uniform VOC patterns; and (iii) cultural diagnosis can be accelerated by taking specific VOC profiles into account. PMID:27604146

  2. Mycobacterium avium paratuberculosis and the etiology of Crohn's disease: a review of the controversy from the clinician's perspective.

    Science.gov (United States)

    Rosenfeld, Greg; Bressler, Brian

    2010-10-01

    Mycobacterium avium paratuberculosis (MAP) is an obligate intracellular organism that has frequently been associated with Crohn's disease (CD). Because CD is a chronic inflammatory condition, many researchers have speculated that an infectious agent must be the cause of CD. MAP has often been proposed to be one such agent; however, despite considerable research, the evidence remains inconclusive. Higher levels of MAP have been found in the tissues and blood of CD patients than in controls, forming the foundation for much of the research into the role of MAP in CD and the primary argument in support of a causative role for MAP in CD. MAP is a slow-growing and fastidious organism that is difficult to grow in culture and, therefore, challenging to detect in patients. As a result, there has been variability in the results of studies attempting to detect the presence of MAP in CD patients, and considerable controversy over whether this organism has a causative role in the etiology of CD. Two main hypotheses exist with respect to the role of MAP in CD. The first is that MAP is a principal cause of CD, while the second is that MAP is more prevalent because of the immune dysfunction seen in CD but does not play a causative role. Clinicians are often faced with questions regarding the role of this organism and the need to treat it. The present article attempts to provide an overview of the controversy including the nature of the mycobacterium, the difficulty in detecting it, the use of antimycobacterial agents to treat it and the effect of immunosuppressive agents - all from a clinician's perspective. Although the role of MAP in CD remains controversial and an area of considerable research, it is currently only of academic interest because there is no clinically useful test to identify the presence of the organism, and no evidence to support the use of antibiotics to eradicate it for the treatment of CD. PMID:21037992

  3. Systems biology analysis of gene expression during in vivo Mycobacterium avium paratuberculosis enteric colonization reveals role for immune tolerance.

    Directory of Open Access Journals (Sweden)

    Sangeeta Khare

    Full Text Available Survival and persistence of Mycobacterium avium subsp. paratuberculosis (MAP in the intestinal mucosa is associated with host immune tolerance. However, the initial events during MAP interaction with its host that lead to pathogen survival, granulomatous inflammation, and clinical disease progression are poorly defined. We hypothesize that immune tolerance is initiated upon initial contact of MAP with the intestinal Peyer's patch. To test our hypothesis, ligated ileal loops in neonatal calves were infected with MAP. Intestinal tissue RNAs were collected (0.5, 1, 2, 4, 8 and 12 hrs post-infection, processed, and hybridized to bovine gene expression microarrays. By comparing the gene transcription responses of calves infected with the MAP, informative complex patterns of expression were clearly visible. To interpret these complex data, changes in the gene expression were further analyzed by dynamic Bayesian analysis, and genes were grouped into the specific pathways and gene ontology categories to create a holistic model. This model revealed three different phases of responses: i early (30 min and 1 hr post-infection, ii intermediate (2, 4 and 8 hrs post-infection, and iii late (12 hrs post-infection. We describe here the data that include expression profiles for perturbed pathways, as well as, mechanistic genes (genes predicted to have regulatory influence that are associated with immune tolerance. In the Early Phase of MAP infection, multiple pathways were initiated in response to MAP invasion via receptor mediated endocytosis and changes in intestinal permeability. During the Intermediate Phase, perturbed pathways involved the inflammatory responses, cytokine-cytokine receptor interaction, and cell-cell signaling. During the Late Phase of infection, gene responses associated with immune tolerance were initiated at the level of T-cell signaling. Our study provides evidence that MAP infection resulted in differentially regulated genes, perturbed

  4. Evaluation of a Salmonella vectored vaccine expressing Mycobacterium avium subsp. paratuberculosis antigens against challenge in a goat model.

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    Syed M Faisal

    Full Text Available Johnes disease (JD, caused by Mycobacterium avium subsp paratuberculosis (MAP, occurs worldwide as chronic granulomatous enteritis of domestic and wild ruminants. To develop a cost effective vaccine, in a previous study we constructed an attenuated Salmonella strain that expressed a fusion product made up of partial fragments of MAP antigens (Ag85A, Ag85B and SOD that imparted protection against challenge in a mouse model. In the current study we evaluated the differential immune response and protective efficacy of the Sal-Ag vaccine against challenge in a goat model as compared to the live attenuated vaccine MAP316F. PBMCs from goats vaccinated with Sal-Ag and challenged with MAP generated significantly lower levels of IFN-γ, following in vitro stimulation with either Antigen-mix or PPD jhonin, than PBMC from MAP316F vaccinated animals. Flow cytometric analysis showed the increase in IFN-γ correlated with a significantly higher level of proliferation of CD4, CD8 and γδT cells and an increased expression of CD25 and CD45R0 in MAP316F vaccinated animals as compared to control animals. Evaluation of a range of cytokines involved in Th1, Th2, Treg, and Th17 immune responses by quantitative PCR showed low levels of expression of Th1 (IFN-γ, IL-2, IL-12 and proinflammatory cytokines (IL-6, IL-8, IL-18, TNF-α in the Sal-Ag immunized group. Significant levels of Th2 and anti-inflammatory cytokines transcripts (IL-4, IL-10, IL-13, TGF-β were expressed but their level was low and with a pattern similar to the control group. Over all, Sal-Ag vaccine imparted partial protection that limited colonization in tissues of some animals upon challenge with wild type MAP but not to the level achieved with MAP316F. In conclusion, the data indicates that Sal-Ag vaccine induced only a low level of protective immunity that failed to limit the colonization of MAP in infected animals. Hence the Sal-Ag vaccine needs further refinement to increase its efficacy.

  5. Validation and standardization of IS900 and F57 real-time quantitative PCR assays for the specific detection and quantification of Mycobacterium avium subsp. paratuberculosis.

    Science.gov (United States)

    Sidoti, Francesca; Banche, Giuliana; Astegiano, Sara; Allizond, Valeria; Cuffini, Anna Maria; Bergallo, Massimiliano

    2011-05-01

    Mycobacterium avium subsp. paratuberculosis (Map) is the causative agent of Johne's disease and may contribute to the onset and development of Crohn's disease in humans. Rapid detection of Map is fundamental because of its reported isolation from pasteurized milk and its potential for transmission through environmental sources. In this study, we developed two independent real-time quantitative PCR assays targeting the IS900 genetic insertion sequence and the F57 sequence, which proved capable of detecting and quantifying Map DNA. Validation and standardization of the developed methods were performed by evaluating diagnostic trueness, precision, and accuracy of the techniques. Specificity of the IS900 and F57 methods was verified in both in silico and experimental studies. The assays were found to be very accurate and precise with high repeatability and reproducibility. Moreover, the two real-time assays were very specific for Map, discriminating most of mycobacterial and nonmycobacterial species. PMID:21510779

  6. Development of a novel DNA extraction method for identification and quantification of Mycobacterium avium subsp. paratuberculosis from tissue samples by real-time PCR.

    Science.gov (United States)

    Park, Kun Taek; Allen, Andrew J; Davis, William C

    2014-04-01

    Mycobacterium avium subsp. paratuberculosis (Map) is the causative agent of Johne's disease in ruminants and possibly associated with human Crohn's disease. One impediment in furthering our understanding of this potential association has been the lack of an accurate method for detection of Map in affected tissues. Real time polymerase chain reaction (RT-PCR) methods have been reported to have different sensitivities in detection of Map. This is in part attributable to the difficulties of extracting Map DNA and removing PCR inhibitors from the clinical specimens. The maximum efficiency of RT-PCR can only be achieved by using high quality DNA samples. In this study, we present a novel pre-treatment method which significantly increases Map DNA recovery and decreases PCR inhibitors (pMap DNA extraction method from tissue samples will make RT-PCR a more powerful tool for a wide range of applications for Map identification and quantification. PMID:24534783

  7. Effects of fractionated colostrum replacer and vitamins A, D, and E on haptoglobin and clinical health in neonatal Holstein calves challenged with Mycobacterium avium ssp. paratuberculosis.

    Science.gov (United States)

    Krueger, L A; Reinhardt, T A; Beitz, D C; Stuart, R L; Stabel, J R

    2016-04-01

    Thirty Holstein calves were obtained from 2 dairy farms in central Iowa at birth and randomly assigned to 1 of 6 treatment groups: (1) colostrum deprived (CD), no vitamins; (2) colostrum replacer (CR), no vitamins; (3) CR, vitamin A; (4) CR, vitamin D3; (5) CR, vitamin E; and (6) CR, vitamins A, D3, E, with 5 calves per treatment in a 14-d study. Calves were fed pasteurized whole milk (CD) or fractionated colostrum replacer (CR) at birth (d 0) and injected with vitamins according to treatment group. From d 1 through d 14 of the study, all calves were fed pasteurized whole milk (PWM) supplemented with vitamins as assigned. All calves were inoculated with Mycobacterium avium ssp. paratuberculosis on d 1 and 3 of age. Calves fed CR acquired IgG1 and haptoglobin in serum within 24 h of birth, whereas CD calves did not. The CR-fed calves were 2.5 times less likely to develop scours, and CR calves supplemented with vitamins D3 and E also demonstrated a decreased incidence of scours. Serum vitamin levels of A, D, and E increased within treatment group by d 7 and 14 of the study. Interestingly, synergistic effects of supplemental vitamins A, D3, and E on serum 25-(OH)-vitamin D were observed at d 7, resulting in higher levels than in calves administered vitamin D only. Further, vitamin D3 deficiency was observed in CD and CR calves fed a basal diet of pasteurized whole milk and no supplemental vitamins. Colonization of tissues with Mycobacterium avium ssp. paratuberculosis was negligible and was not affected by colostrum feeding or vitamin supplementation. Results demonstrated passive transfer of haptoglobin to neonatal calves, and potential health benefits of supplemental vitamins D3 and E to calves fed pasteurized whole milk. PMID:26805975

  8. Characterization of the Mycobacterium avium subsp. paratuberculosis laminin-binding/histone-like protein (Lbp/Hlp) which reacts with sera from patients with Crohn's disease.

    Science.gov (United States)

    Lefrançois, Louise H; Pujol, Céline; Bodier, Christelle C; Teixeira-Gomez, Ana Paula; Drobecq, Hervé; Rosso, Marie-Laure; Raze, Dominique; Dias, André Alves; Hugot, Jean-Pierre; Chacon, Ofelia; Barletta, Raul G; Locht, Camille; Vidal Pessolani, Maria Cristina; Biet, Franck

    2011-06-01

    Mycobacterium avium subsp. paratuberculosis (Map) causes a chronic enteric disease in ruminants, called paratuberculosis or Johne's disease. The current model proposes that after ingestion by the host, Map crosses the intestinal barrier via internalization by the M cells. Experimental observations suggest, however, that Map may also transcytose the intestinal wall via the enterocytes, but the mechanisms involved in this process remain poorly understood. Cytoadherence assays performed on epithelial cells with Map revealed that the addition of laminin to the cell culture increases adhesion. A Map protein was isolated by heparin-Sepharose chromatography and identified as a laminin-binding protein like. The gene encoding this protein named Lbp/Hlp was identified in the Map genome sequence at locus MAP3024 (annotated Hup B). The deduced Map Lbp/Hlp amino acid sequence reveals 80% identity with that reported for other mycobacteria. The C-terminal domain involved in adhesion is mainly composed of arginine and lysine residues modified by methylation. In vitro tests demonstrated that recombinant Lbp/Hlp binds laminin, heparin, collagen and epithelial cells. Interestingly, we found that this adhesin corresponds to the antigen described as the target of pANCA and serum antibodies of patients with Crohn's disease. PMID:21334452

  9. Anti-Inflammatory and Antiapoptotic Responses to Infection: A Common Denominator of Human and Bovine Macrophages Infected with Mycobacterium avium Subsp. paratuberculosis

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    Naiara Abendaño

    2013-01-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (Map is the causative agent of a chronic intestinal inflammation in ruminants named Johne's disease or paratuberculosis and a possible etiopathological agent of human Crohn's disease (CD. Analysis of macrophage transcriptomes in response to Map infection is expected to provide key missing information in the understanding of the role of this pathogen in establishing an inappropriate and persistent infection in a susceptible host and of the molecular mechanisms that might underlie the early phases of CD. In this paper we summarize transcriptomic studies of human and bovine peripheral blood mononuclear cells (PBMC, monocyte-derived macrophages (MDMs, and macrophages-like cell lines in vitro infected with Map. Most studies included in this paper consistently reported common gene expression signatures of bovine and human macrophages in response to Map such as enhanced expression of the anti-inflammatory cytokines IL-10 and IL-6, which promote bacterial survival. Overexpression of IL-10 could be responsible for the Map-associated reduction in the expression of the proapoptotic TNF-α gene observed in bovine and human macrophages.

  10. Mycobacterium avium Subsp. avium Infection in Four Veal Calves: Differentiation from Intestinal Tuberculosis

    OpenAIRE

    2014-01-01

    Mycobacterium avium subsp. avium (Maa) is an intracellular pathogen belonging to the Mycobacterium avium-intracellulare complex (MAC). Reservoirs of MAC are the natural environment, wildlife and domestic animals. In adult bovine, MAC infections are typically caused by Mycobacterium avium subsp. paratuberculosis (Map). Maa infections in bovine are rarely reported but may cause clinical disease and pathological lesions similar to those observed in paratuberculosis or those induced by members of...

  11. 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

  12. Effectiveness of combination of Mini-and Microsatellite loci to sub-type Mycobacterium avium subsp. paratuberculosis Italian type C isolates

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    Garbarino Chiara A

    2011-09-01

    Full Text Available Abstract Background Mycobacterium avium subsp. paratuberculosis (Map is the etiological agent of paratuberculosis. The aim of our study was to combine Mini-and Microsatellite loci analysis in order to explore the effectiveness of this sub-typing method in a group of Map isolates. For this purpose, 84 Italian Type C Map isolates, each from a different cattle herd, were submitted to MIRU-Variable-Number Tandem-Repeats (VNTRs typing and Short Sequence repeats (SSRs sequencing. Moreover, the method was used to analyse the variability inside 10 herds (from three to 50 isolates per herd. Results The molecular sub-typing, carried out using three SSR and 10 MIRU-VNTR loci, differentiated the 84 isolates into 33 clusters, reaching a Simpson's Discriminatory Index (SID value of 0.952 (0.933 to 0.972, 95% confidence intervals. Among all considered loci, six (SSR2, MIRU2, SSR1, SSR8, VNTR3527 and VNTR1067 showed relevant allelic variability. Thirty-eight% of the isolates were clustered into four genotypes, differing from each other for the SSR2 locus. The other isolates, characterised by differences in two or more loci, were spread among the rest of the clusters. The intra-herd analysis revealed more than one genotype in most herds with a similar distribution of clusters. Conclusions Our results revealed the advantage of using both Mini-and Microsatellite approaches for successfully discriminating among Map Type C isolates from the same geographic area, host species and herd. These data suggest that the combination of loci here proposed could be a useful molecular tool for regional epidemiological studies.

  13. Detection and confirmation of Mycobacterium avium subsp. paratuberculosis in direct quantitative PCR positive fecal samples by the manual fluorescent MGIT culture system.

    Science.gov (United States)

    Kawaji, Satoko; Nagata, Reiko; Mori, Yasuyuki

    2014-01-01

    An efficient protocol for the manual fluorescent MGIT culture system combined with rapid confirmation of Mycobacterium avium subsp. paratuberculosis (MAP) growth in the broth culture was established and evaluated for the detection of viable MAP in direct quantitative PCR (QPCR) positive bovine feces. Manually detected fluorescence emissions from MGIT tubes were analyzed objectively using an open source software, ImageJ. For molecular confirmation of MAP growth, DNA samples harvested by simply boiling the broth, an inexpensive and time- and labor-saving DNA preparation method, yielded adequate results. The sheep strain of MAP required longer incubation time relative to the cattle strain, suggesting that the MGIT system may not support well the growth of ovine isolates as described previously. Of 61 direct QPCR positive bovine feces, the recovery rate of MAP in the MGIT system (62.3%) was significantly higher (Pculture by the MGIT system was several weeks earlier compared to solid media. In MGIT culture positive samples, the time to detect fluorescence was correlated with the DNA quantity detected in fecal QPCR. As a positive result in the direct fecal QPCR test does not mean fecal excretion of viable MAP, bacterial isolation by fecal culture could be conducted to verify the QPCR result. For this purpose, the manual MGIT system is a sensitive and rapid culture method at least for bovine samples. PMID:24065085

  14. Repeated cycles of chemical and physical disinfection and their influence on Mycobacterium avium subsp. paratuberculosis viability measured by propidium monoazide F57 quantitative real time PCR.

    Science.gov (United States)

    Kralik, Petr; Babak, Vladimir; Dziedzinska, Radka

    2014-09-01

    Mycobacterium avium subsp. paratuberculosis (MAP) has a high degree of resistance to chemical and physical procedures frequently used for the elimination of other bacteria. Recently, a method for the determination of viability by exposure of MAP to propidium monoazide (PMA) and subsequent real time quantitative PCR (qPCR) was established and found to be comparable with culture. The aim of this study was to apply the PMA qPCR method to determine the impact of increasing concentration or time and repeated cycles of the application of selected disinfectants on MAP viability. Different MAP isolates responded to the same type of stress in different ways. The laboratory strain CAPM 6381 had the highest tolerance, while the 8819 low-passage field isolate was the most sensitive. Ultraviolet exposure caused only a partial reduction in MAP viability; all MAP isolates were relatively resistant to chlorine. Only the application of peracetic acid led to the total elimination of MAP. Repeated application of the treatments resulted in more significant decreases in MAP viability compared to single increases in the concentration or time of exposure to the disinfectant. PMID:24934261

  15. Molecular analysis and MIRU-VNTR typing of Mycobacterium avium subsp. avium, 'hominissuis' and silvaticum strains of veterinary origin.

    Science.gov (United States)

    Rónai, Zsuzsanna; Csivincsik, Ágnes; Dán, Ádám; Gyuranecz, Miklós

    2016-06-01

    Besides Mycobacterium avium subsp. paratuberculosis (MAP), M. avium subsp. avium (MAA), M. avium subsp. silvaticum (MAS), and 'M. avium subsp. hominissuis' (MAH) are equally important members of M. avium complex, with worldwide distribution and zoonotic potential. Genotypic discrimination is a prerequisite to epidemiological studies which can facilitate disease prevention through revealing infection sources and transmission routes. The primary aim of this study was to identify the genetic diversity within 135 MAA, 62 MAS, and 84 MAH strains isolated from wild and domestic mammals, reptiles and birds. Strains were tested for the presence of large sequence polymorphism LSP(A)17 and were submitted to Mycobacterial interspersed repetitive units-variable-number tandem repeat (MIRU-VNTR) analysis at 8 loci, including MIRU1, 2, 3, and 4, VNTR25, 32, and 259, and MATR9. In 12 strains hsp65 sequence code type was also determined. LSP(A)17 was present only in 19.9% of the strains. All LSP(A)17 positive strains belonged to subspecies MAH. The discriminatory power of the MIRU-VNTR loci set used reached 0.9228. Altogether 54 different genotypes were detected. Within MAH, MAA, and MAS strains 33, 16, and 5 different genotypes were observed. The described genotypes were not restricted to geographic regions or host species, but proved to be subspecies specific. Our knowledge about MAS is limited due to isolation and identification difficulties. This is the first study including a large number of MAS field strains. Our results demonstrate the high diversity of MAH and MAA strains and the relative uniformity of MAS strains. PMID:26964909

  16. 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

    genotype. Cows with the TLR2-1903 T/C mutation (i.e., the CT and CC genotypes) were at 1.7 (95% CI: 1.2, 2.8) times the odds of being Map-infected compared to cows with the TT genotype. In in vitro functional assays, monocyte-derived macrophages from animals with a TLR2-1903 TT genotype produced more IL12p......40 and IL1β when stimulated with Map compared to cells derived from TLR2-1903 CT and CC genotypes. Also, T cell proliferative responses to mycobacterial antigens were higher in animals with a TLR2-1903 TT genotype. In conclusion, we have found a significant association between SNP TLR2-1903 T...

  17. Dairy farms testing positive for Mycobacterium avium ssp. paratuberculosis have poorer hygiene practices and are less cautious when purchasing cattle than test-negative herds.

    Science.gov (United States)

    Wolf, R; Barkema, H W; De Buck, J; Orsel, K

    2016-06-01

    Mycobacterium avium ssp. paratuberculosis (MAP), the causative agent of Johne's disease, is present on most dairy farms in Alberta, causing economic losses and presenting a potential public health concern. The objective of this cross-sectional study was to identify risk factors for Alberta dairy herds being MAP-positive based on environmental samples (ES). Risk assessments were conducted and ES were collected on 354 Alberta dairy farms (62% of eligible producers) voluntarily participating in the Alberta Johne's Disease Initiative. In univariate logistic regression, risk factors addressing animal and pen hygiene, as well as the use of feeding equipment to remove manure and manure application on pastures, were all associated with the number of positive ES. Furthermore, based on factor analysis, risk factors were clustered and could be summarized as 4 independent factors: (1) animal, pen, and feeder contamination; (2) shared equipment and pasture contamination; (3) calf diet; and (4) cattle purchase. Using these factor scores as independent variables in multivariate logistic regression models, a 1-unit increase in animal, pen, and feeder contamination resulted in 1.31 times higher odds of having at least 1 positive ES. Furthermore, a 1-unit increase in cattle purchase also resulted in 1.31 times the odds of having at least 1 positive ES. Finally, a 100-cow increase in herd size resulted in an odds ratio of 2.1 for having at least 1 positive ES. In conclusion, cleanliness of animals, pens, and feeders, as well as cattle purchase practices, affected risk of herd infection with MAP. Therefore, improvements in those management practices should be the focus of effective tools to control MAP on dairy farms. PMID:26995127

  18. Association of single nucleotide polymorphisms in the ANKRA2 and CD180 genes with bovine respiratory disease and presence of Mycobacterium avium subsp. paratuberculosis(1).

    Science.gov (United States)

    Casas, E; Garcia, M D; Wells, J E; Smith, T P L

    2011-12-01

    The objective was to determine whether single nucleotide polymorphisms (SNPs) in the ANKRA2 and CD180 genes are associated with incidence of bovine respiratory disease (BRD) and presence of Mycobacterium avium subsp. paratuberculosis (MAP) in cattle. Two independent populations were used. The first population (BRD-affected; N = 90) was composed of 31 half-sib progeny, from a Brahman × Angus sire, that were treated for BRD. Untreated offspring from the sire were selected to serve as controls. The second population (MAP-infected) of 330 animals of unknown parentage was evaluated for the presence of MAP in ileocecal lymph node and classified as positive or negative. Markers in both genes were assessed for association in these two populations. In the BRD-affected population, five SNPs in the ANKRA2 gene were significantly associated (P < 0.05), and two SNPs were highly associated (P < 0.01) with incidence of BRD. In addition, two SNPs in the CD180 gene were found to be associated with this trait. In the MAP-infected population, one SNP in the ANKRA2 gene was significantly associated (P < 0.05) with the presence or absence of MAP, and a SNP in the CD180 gene was highly associated (P < 0.01) with the trait. Haplotypes, using significant markers, showed a positive association with both incidence of BRD (P = 0.0001) and with the presence of MAP (P = 0.0032). Markers in the ANKRA2 and CD180 genes are associated with the ability of the animal to cope with pathogens. PMID:22034997

  19. Estimation of the prevalence at animal level of paratuberculosis in female cattle of Saxony-Anhalt (Germany

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    Nicolai Denzin

    2011-11-01

    Full Text Available Johne’s disease (paratuberculosis is a chronic, untreatable disease of ruminants caused by Mycobacterium avium subspecies paratuberculosis. It leads to economic losses in livestock production but is also suspected to be involved in the pathogenesis of Crohn’s disease in humans. In 2007/2008 a study was conducted to estimate the prevalence at animal level of paratuberculosis among the female cattle of Saxony-Anhalt (Germany. In order to keep study costs low and participating farms confidential, no farms were visited to actively draw samples. Instead, animals were pre-selected in a formal random sampling process and additionally tested by ELISA for paratuberculosis when samples suitable for serology were routinely submitted to the State Office of Consumer Protection Saxony-Anhalt for any diagnostics. Out of 896 samples, 38 tested positive, giving an apparent prevalence of 4.2%. Based on the test quality traits of the ELISA employed in the study (POURQUIER® ELISA Paratuberculosis the true prevalence was estimated to 6.7% (95% CI, 3.0-10.4%. No spatial clustering of positive results was detected by a scan statistic. Prevalence estimates for age strata tended to show an incline to a maximum at age class of 5 years and a subsequent decline with higher age classes. Estimates tended to show an incline with herd size.

  20. Identification and characterization of a spore-like morphotype in chronically starved Mycobacterium avium subsp. paratuberculosis cultures.

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    Elise A Lamont

    Full Text Available Mycobacteria are able to enter into a state of non-replication or dormancy, which may result in their chronic persistence in soil, aquatic environments, and permissive hosts. Stresses such as nutrient deprivation and hypoxia provide environmental cues to enter a persistent state; however, a clear definition of the mechanism that mycobacteria employ to achieve this remains elusive. While the concept of sporulation in mycobacteria is not novel, it continues to spark controversy and challenges our perceptions of a non-replication. We investigated the potential role of sporulation in one-year old broth cultures of Mycobacterium subsp. paratuberculosis (MAP. We show that dormant cultures of MAP contain a mix of vegetative cells and a previously unknown morphotype resembling a spore. These spore-like structures can be enriched for using sporulating media. Furthermore, purified MAP spore forms survive exposure to heat, lysozyme and proteinase K. Heat-treated spores are positive for MAP 16SrRNA and IS900. MAP spores display enhanced infectivity as well as maintain acid-fast characteristics upon germination in a well-established bovine macrophage model. This is the first study to demonstrate a new MAP morphotype possessing spore-like qualities. Data suggest that sporulation may be a viable mechanism by which MAP accomplishes persistence in the host and/or environment. Thus, our current understanding of mycobacterial persistence, pathogenesis, epidemiology and rational drug and vaccine design may need to be reevaluated.

  1. Herd-level prevalence and associated risk factors for Mycobacterium avium subsp. paratuberculosis in cattle in the State of Paraíba, Northeastern Brazil.

    Science.gov (United States)

    Vilar, Ana L T; Santos, Carolina S A B; Pimenta, Carla L R M; Freitas, Theonys D; Brasil, Arthur W L; Clementino, Inácio J; Alves, Clebert J; Bezerra, Camila S; Riet-Correa, Franklin; Oliveira, Taynara S; Azevedo, Sérgio S

    2015-09-01

    A cross-sectional study based on a planned sampling was carried out to determine herd-level and animal-level prevalences, and to identify risk factors associated with herd-level prevalence for bovine paratuberculosis in the State of Paraíba, Northeastern Brazil. The state was divided into three sampling groups: sampling stratum 1 (mesoregion of Sertão), sampling stratum 2 (mesoregion of Borborema), and sampling stratum 3 (mesoregions of Zona da Mata and Agreste). For each sampling stratum, herd-level and animal-level prevalences were estimated by a two-stage sampling survey. In the first stage, a pre-established number of herds (primary sampling units) were randomly selected; in the second stage, a pre-established number of cows aged ≥24 months were randomly selected (secondary sampling units). Ten animals were sampled in herds with up to 99 cows aged over 24 months; 15 animals were sampled in herds with 100 or more cows aged over 24 months; and all animals were sampled in those with up to 10 cows aged over 24 months. In total, 2504 animals were sampled from 480 herds. Enzyme-linked immunosorbant assay (ELISA) test kits were used for Mycobacterium avium subsp. paratuberculosis (MAP) antibody detection. A herd was deemed positive for the presence of MAP if it included at least one positive animal in herds of up to 24 females, and two positive animals in herds with more than 24 females. The herd-level prevalence in the State of Paraíba was 34.5% (95% CI=30.2-39.1%), 26.6% (95% CI=20.2-34.2%) in the region of Borborema, 30.5% (95% CI=23.9-38.0%) in Agreste/Mata, and 41.4% (95% CI=34.0-49.1%) in Sertão. The animal-level prevalence was 10.7% (95% CI=7.3-15.4%) in the State of Paraíba, 7.9% (95% CI=5.2-11.7%) in the region of Borborema, 9.4% (95% CI=7.3-12.1%) in Sertão, and 13.9% (95% CI=6.2--28.3%) in Agreste/Mata. The frequency of seropositive animals per herd ranged from 6.7% to 100% (median of 20%). The risk factors identified were as follows: Sertão region

  2. Evaluation of testing strategies to identify infected animals at a single round of testing within dairy herds known to be infected with Mycobacterium avium ssp. paratuberculosis.

    Science.gov (United States)

    More, S J; Cameron, A R; Strain, S; Cashman, W; Ezanno, P; Kenny, K; Fourichon, C; Graham, D

    2015-08-01

    As part of a broader control strategy within herds known to be infected with Mycobacterium avium ssp. paratuberculosis (MAP), individual animal testing is generally conducted to identify infected animals for action, usually culling. Opportunities are now available to quantitatively compare different testing strategies (combinations of tests) in known infected herds. This study evaluates the effectiveness, cost, and cost-effectiveness of different testing strategies to identify infected animals at a single round of testing within dairy herds known to be MAP infected. A model was developed, taking account of both within-herd infection dynamics and test performance, to simulate the use of different tests at a single round of testing in a known infected herd. Model inputs included the number of animals at different stages of infection, the sensitivity and specificity of each test, and the costs of testing and culling. Testing strategies included either milk or serum ELISA alone or with fecal culture in series. Model outputs included effectiveness (detection fraction, the proportion of truly infected animals in the herd that are successfully detected by the testing strategy), cost, and cost-effectiveness (testing cost per true positive detected, total cost per true positive detected). Several assumptions were made: MAP was introduced with a single animal and no management interventions were implemented to limit within-herd transmission of MAP before this test. In medium herds, between 7 and 26% of infected animals are detected at a single round of testing, the former using the milk ELISA and fecal culture in series 5 yr after MAP introduction and the latter using fecal culture alone 15 yr after MAP introduction. The combined costs of testing and culling at a single round of testing increases with time since introduction of MAP infection, with culling costs being much greater than testing costs. The cost-effectiveness of testing varied by testing strategy. It was also

  3. Seroreactivity against Specific L5P Antigen from Mycobacterium avium subsp. paratuberculosis in Children at Risk for T1D

    Science.gov (United States)

    Niegowska, Magdalena; Rapini, Novella; Biet, Frank; Piccinini, Simona; Bay, Sylvie; Lidano, Roberta; Manca Bitti, Maria Luisa; Sechi, Leonardo A.

    2016-01-01

    Aims/Hypothesis Although numerous environmental agents have been investigated over the years as possible triggers of type 1 diabetes (T1D), its causes remain unclear. We have already demonstrated an increased prevalence of antibodies against peptides derived from Mycobacterium avuim subsp. paratuberculosis (MAP) homologous to human zinc transporter 8 protein (ZnT8) and proinsulin in Italian subjects at risk for or affected by T1D. In this study, we compared titers of the previously detected antibodies with seroreactivity to MAP lipopentapetide (L5P) that recently emerged as a strong immunogenic component able to specifically distinguish MAP from other mycobacteria. Methods Plasma of 32 children and youth at risk for T1D including follow-up samples and 42 age-matched healthy controls (HC) recruited at the Tor Vergata University Hospital in Rome was analyzed by indirect ELISA for the presence of antibodies against MAP-derived epitopes MAP3865c133–141, MAP3865c125-133, MAP2404c70-85 and MAP1,4αgbp157-173 along with their ZnT8 and proinsulin homologs. The data were analyzed through two-tailed Mann-Whitney U test and relation between variables was determined by principal component analysis. Results Responses to L5P were not detectable in subjects whose initial seroreactivity to MAP peptides and their human homologs was lost in follow-up samples, whereas anti-L5P antibodies appeared constantly in individuals with a stable immunity against MAP antigens. The overall coincidence in positivity to L5P and the four MAP epitopes both in children at risk for T1D and HC exceeded 90%. Conclusions MAP-derived homologs may cross-react with ZnT8 and proinsulin peptides inducing immune responses at a young age in subjects predisposed for T1D. Thus, L5P may have a diagnostic value to immediately indicate the presence of anti-MAP seroreactivity when evaluation of a more complex antibody status is not required. Almost complete coincidence in responses to both types of antigens lends

  4. Seroreactivity against Specific L5P Antigen from Mycobacterium avium subsp. paratuberculosis in Children at Risk for T1D.

    Directory of Open Access Journals (Sweden)

    Magdalena Niegowska

    Full Text Available Although numerous environmental agents have been investigated over the years as possible triggers of type 1 diabetes (T1D, its causes remain unclear. We have already demonstrated an increased prevalence of antibodies against peptides derived from Mycobacterium avuim subsp. paratuberculosis (MAP homologous to human zinc transporter 8 protein (ZnT8 and proinsulin in Italian subjects at risk for or affected by T1D. In this study, we compared titers of the previously detected antibodies with seroreactivity to MAP lipopentapetide (L5P that recently emerged as a strong immunogenic component able to specifically distinguish MAP from other mycobacteria.Plasma of 32 children and youth at risk for T1D including follow-up samples and 42 age-matched healthy controls (HC recruited at the Tor Vergata University Hospital in Rome was analyzed by indirect ELISA for the presence of antibodies against MAP-derived epitopes MAP3865c133-141, MAP3865c125-133, MAP2404c70-85 and MAP1,4αgbp157-173 along with their ZnT8 and proinsulin homologs. The data were analyzed through two-tailed Mann-Whitney U test and relation between variables was determined by principal component analysis.Responses to L5P were not detectable in subjects whose initial seroreactivity to MAP peptides and their human homologs was lost in follow-up samples, whereas anti-L5P antibodies appeared constantly in individuals with a stable immunity against MAP antigens. The overall coincidence in positivity to L5P and the four MAP epitopes both in children at risk for T1D and HC exceeded 90%.MAP-derived homologs may cross-react with ZnT8 and proinsulin peptides inducing immune responses at a young age in subjects predisposed for T1D. Thus, L5P may have a diagnostic value to immediately indicate the presence of anti-MAP seroreactivity when evaluation of a more complex antibody status is not required. Almost complete coincidence in responses to both types of antigens lends support to the involvement of MAP

  5. The Occurrence of Paratuberculosis (Johne’s Disease in Ruminants in Indonesia Must be Anticipated

    Directory of Open Access Journals (Sweden)

    Tarmudji

    2007-06-01

    Full Text Available Paratuberculosis or Johne’s disease is an infectious disease in ruminants (cattle, buffalo, sheep and goat caused by Mycobacterium avium subspecies paratuberculosis (MAP and characterized by granulomatous enteritis manifestation. The disease occurs worldwidely and causes great economic losses on domestic livestock industries. Calves are commonly infected soon after birth, with incubation period of either some months or years. Clinical signs observed from 2 to 10 years old of infected cattle are chronic diarrhea and progressive emaciation. Transmission of MAP to calves can occur by nursing the infected dam or got contaminated by fecal material. The pathogens can also be excreted in colostrum or milk, that is why calf can be infected since neonatal period. Infection in progress leads to cause thickening of the intestinal wall, granulomatous and mesenterical lymphnode, which diffusion lesions in the intestine are characterized by the macroscopical finding. In Indonesia, paratuberculosis had been reported in dairy cattle (in West Java with seroprevalence of 1.67% (3/180. From the serological positive reactors demonstrated MAP of 0.55% (1/180 by fecal cuture examination. Some samples of cattle and buffaloes from North Sumatera were also found positive paratuberculosis antibody against MAP detected by Complement Fixation Test (CFT at average of 4% (2/50. The presence of positive reactors of paratuberculosis in dairy cattle, beef cattle and buffaloes in Indonesia must be anticipated. These animals are carriers and can shed pathogens, although they do not show clinical signs. It is likely that paratuberculosis can not be detected by conventional diagnostic techniques, therefore, sensitive and early diagnosis techniques must be developed.

  6. Effect of days in milk and milk yield on testing positive in milk antibody ELISA to Mycobacterium avium subsp. paratuberculosis in dairy cattle

    DEFF Research Database (Denmark)

    Nielsen, Søren Saxmose; Toft, Nils

    2012-01-01

    Milk samples are becoming more used as a diagnostic specimen for assessment of occurrence of antibodies to Mycobacterium avium subsp. paratuberculosis (MAP). This study assessed the effect of days in milk (DIM) and milk yield on testing positive in a commercial MAP specific milk antibody ELISA...... among 222,774 Danish Holstein cows. Results showed that odds of testing positive on 1-2 DIM were 9-27 times higher than the rest of lactation, where the chance of testing positive varied less. The reason is most likely a high concentration of non-specific antibodies in colostrum. Consequently, samples...... from the first couple of DIM should be excluded from MAP testing until further information on their significance is established. Milk yield also had a significant effect on odds of testing positive due to its diluting effect. Inclusion of milk yield in the interpretation of test results could improve...

  7. Avaliação sorológica e de fatores de risco para a infecção por Mycobacterium avium subsp. paratuberculosis em rebanhos leiteiros da Microrregião de Garanhuns, Pernambuco Serological evaluation and risk factors for Mycobacterium avium subsp. paratuberculosis infection in dairy herds of Microregion Garanhuns, Pernambuco

    Directory of Open Access Journals (Sweden)

    Luenda de M. e Sá

    2013-03-01

    Full Text Available Objetivou-se com esse trabalho realizar um inquérito epidemiológico da infecção por Mycobacterium avium subsp. paratuberculosis (MAP em bovinos leiteiros da microrregião de Garanhuns, Pernambuco, Brasil. Para este estudo foram coletadas amostras sanguíneas de 408 animais, provenientes de 19 rebanhos localizados em 15 municípios. O exame sorológico foi realizado por Ensaio Imunoenzimático (ELISA indireto para detecção de anticorpos frente ao MAP. Em todas as propriedades, foi aplicado um questionário investigativo para análise dos fatores de risco, e as coordenadas geográficas coletadas por um aparelho de Global Position System (GPS para realização da distribuição espacial. A prevalência da infecção por MAP foi de 2,7% (11/408; I.C. 1,4-4,9. O número de focos foi 47,4% (9/19. Na análise de regressão logística foi identificado como fator de risco a taxa anual de nascimentos superior a 51 bezerros/ano (OR 3,8; I.C. 1,1-13,1. Desta forma, conclui-se que a infecção por MAP encontra-se presente nos rebanhos bovinos leiteiros da microrregião estudada e que medidas de controle baseadas nos fatores de risco identificados devem ser implementadas com o objetivo de reduzir o número de focos da infecção.The present study aimed to conduct an epidemiological investigation of Mycobacterium avium subsp. paratuberculosis (MAP infection in dairy cattle of the Garanhuns microregion, in Pernambuco, Brazil. Blood samples were collected from 408 animals from 19 herds located in 15 cities. Serological tests were performed by indirect immunoenzymatic assay (ELISA for antibodies against MAP. In all farms, a questionnaire to investigate risk factors was used, and Global Position System (GPS receivers were used to collect geographic coordinates to show the spatial distribution of the animals. The prevalence of MAP infected cattle was 2.7% (11/408; I.C. 1.4-4.9. The rate of infection was 47.4% (9/19. An annual birth rate over 51 calves

  8. Assessment of Live Candidate Vaccines for Paratuberculosis in Animal Models and Macrophages▿

    OpenAIRE

    Scandurra, Gabriella M.; de Lisle, Geoffrey W.; Cavaignac, Sonia M.; Young, May; Kawakami, R. Pamela; Collins, Desmond M

    2009-01-01

    Mycobacterium avium subsp. paratuberculosis (basonym M. paratuberculosis) is the causative agent of paratuberculosis, a chronic enteritis of ruminants. To control the considerable economic effect that paratuberculosis has on the livestock industry, a vaccine that induces protection with minimal side effects is required. We employed transposon mutagenesis and allelic exchange to develop three potential vaccine candidates, which were then tested for virulence with macrophages, mice, and goats. ...

  9. Isolation and Identification of Mycobacterium avium Subsp. paratuberculosis from Milk, Manure and Fecal Samples of Holstein-Friesian Dairy Cattle

    OpenAIRE

    M. Seyyedin; T.Z. Salehi; H. Tadjbakhsh; M.F. Najafi; Rabbani, M.

    2008-01-01

    The main aim of this study was detection of subclinical forms of Johne’s disease on dairy farms which were suspected of having Johne’s disease or history of it and confirmatory paraclinical of paratuberculosis in them. Also, the isolated bacteria will be used for genotyping and molecular epidemiology of paratuberculosis in the next studies. Samples were gathered of 137 farms in different parts of Razavi Khorasan Province of Iran. There were 505 fresh-fecal and manure samples and 148 raw milk ...

  10. Immunology of Paratuberculosis Infection and Disease

    Science.gov (United States)

    The study of host immune responses to Mycobacterium avium subsp. paratuberculosis (MAP) is complicated by a number of factors, including the protracted nature of the disease and the stealthy nature of the pathogen. Improved tools for the measurement of immunologic responses in ruminant species, par...

  11. Correlation between rpoB gene mutation in Mycobacterium avium subspecies paratuberculosis and clinical rifabutin and rifampicin resistance for treatment of Crohn's disease

    Institute of Scientific and Technical Information of China (English)

    Daniel R Beckler; Sammer Elwasila; George Ghobrial; John F Valentine; Saleh A Naser

    2008-01-01

    AIM:To investigate overlapping regions of the rpoB gene previously involved with rifamycin resistance in M.tubercu/osis and seek correlation between rpoB mutations in clinical MAP strains with susceptibility to RIF and RFB.METHODS:We designed a molecular-based PCR method for the evaluation of rifabutin (RFB) and rifampicin (RIF)resistance based on probable determinant regions within the rpoB gene of MAP,including the 81 bp variable site located between nucleotides 1363 and 1443.The minimum inhibitory concentration (NIC) for RIF was also determined against 11 NAP isolates in attempt to seek correlation with rpoB sequences.RESULTS:We determined that MAP strain 18 had an IVlIC of>30 mg/L and≤5 mg/L for RIF and RFB respectively,and a significant and novel rpoB mutation C1367T,compared to an MIC of≤1.0 mg/L for both drugs in the wild type MAP.The 30-fold increase in the MIC was a direct result of the rpoB mutation C1367T,which caused an amino acid change Thr456 to Ile456 in the drug's binding site.In addition,MAP strain 185 contained five silent rpoB mutations and exhibited an MIC comparable to the wild-type.Moreover,our in vitro selected mutation in MAP strain UCF5 resulted in the generation of a new resistant strain (UCF5-RIF16r)that possessed T1442C rpoB mutation and an MIC>30 mg/L and>10 mg/L for RIF and RFB respectively.Sequencing of the entire rpoB gene in lAP strains UCF4,18,and UCF5-RIF16r revealed an rpoB mutation A2284C further downstream of the 81 bp variable region in UCF4,accounting for observed slight increase in NIC.In addition,no other significant mutations were found in strains 18 and UCF-RIF16r.CONCLUSION:The data clearly illustrates that clinical and in vitro-selected MAP mutants with rpoB mutations result in resistance to RIF and RFB,and that a single amino acid change in the beta subunit may have a significant impact on RIF resistance.Unconventional drug susceptibility testing such as our molecular approach will be beneficial for evaluation of antibiotic effectiveness.This molecular approach may also serve as a model for other drugs used for treatment of MAP infections.

  12. Immunological detection of sheep experimentally infected with strains of Mycobacterium avium subspecies containing insertion sequence IS901/IS902 and a 40 kDa protein

    DEFF Research Database (Denmark)

    Klausen, Joan; Perez, V.; Giese, Steen Bjørck; Marin, J.F.G.; Ahrens, Peter

    A monoclonal antibody raised against a 40 kDa protein present in certain M. avium strains (IS901/IS902 positive) was used for developing a blocking ELISA. Sera from experimentally infected sheep were evaluated by indirect ELISA, AGID and blocking ELISA. The blocking assay proved to be highly spec...

  13. Mycobacterium avium Subsp. avium Infection in Four Veal Calves: Differentiation from Intestinal Tuberculosis

    Directory of Open Access Journals (Sweden)

    Christine Goepfert

    2014-01-01

    Full Text Available Mycobacterium avium subsp. avium (Maa is an intracellular pathogen belonging to the Mycobacterium avium-intracellulare complex (MAC. Reservoirs of MAC are the natural environment, wildlife and domestic animals. In adult bovine, MAC infections are typically caused by Mycobacterium avium subsp. paratuberculosis (Map. Maa infections in bovine are rarely reported but may cause clinical disease and pathological lesions similar to those observed in paratuberculosis or those induced by members of the Mycobacterium tuberculosis complex (MTBC. Therefore, differentiation of MAC from MTBC infection should be attempted, especially if unusual mycobacterial lesions are encountered. Four veal calves from a fattening farm dying with clinical signs of otitis media, fever, and weight loss were submitted for necropsy. Samples from affected organs were taken for histologic investigation, bacteriologic culture, and bacterial specification using PCR. Macroscopic thickening of the intestinal mucosa was induced by granulomatous enteritis and colitis. Intracytoplasmic acid-fast bacteria were detected by Ziehl-Neelsen stains and PCR revealed positive results for Mycobacterium avium subsp. avium. Clinical and pathological changes of Maa infection in veal calves had features of Mycobacterium avium subsp. paratuberculosis and the MTBC. Therefore, Mycobacterium tuberculosis complex infection should be considered in cases of granulomatous enteritis in calves.

  14. Pathology of subclinical paratuberculosis (Johne's Disease) in Awassi sheep with reference to its occurrence in Jordan

    Science.gov (United States)

    Paratuberculosis (Johne’s disease) is an incurable infectious, chronically progressive enteric disease affecting domestic and exotic ruminants. The causative agent is Mycobacterium Avium Paratuberculosis (M Johnei). In this study, the occurrence of subclinical Johne’s disease in Awassi sheep is inve...

  15. Mycobacterium avium subsp. avium and subsp. hominissuis give different cytokine responses after in vitro stimulation of human blood mononuclear cells.

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    Johanna Thegerström

    Full Text Available BACKGROUND: Mycobacterium avium is the principal etiologic agent of non-tuberculous lymphadenitis in children. It is also a known pathogen for birds and other animals. Genetic typing of M. avium isolates has led to a proposal to expand the set of subspecies to include M. avium subsp. hominissuis. Isolates associated with disease in humans belong to this subspecies. METHODOLOGY/PRINCIPAL FINDINGS: Peripheral blood mononuclear cells from six healthy blood donors were stimulated in vitro with ten isolates of M. avium avium and 11 isolates of M. avium hominissuis followed by multiplex bead array quantification of cytokines in supernatants. M. avium hominissuis isolates induced significantly more IL-10 and significantly less IL-12p70, TNF, IFN-γ and IL-17 when compared to M. avium avium isolates. All strains induced high levels of IL-17, but had very low levels of IL-12p70. CONCLUSION/SIGNIFICANCE: The strong association between M. avium subsp. hominissuis and disease in humans and the clear differences in the human immune response to M. avium subsp. hominissuis compared to M. avium subsp. avium isolates, as demonstrated in this study, suggest that genetic differences between M. avium isolates play an important role in the pathogenicity in humans.

  16. Identification of seroreactive proteins in the culture filtrate antigen of Mycobacterium avium ssp. paratuberculosis human isolates to sera from Crohn's disease patients.

    Science.gov (United States)

    Shin, A-Rum; Kim, Hwa-Jung; Cho, Sang Nae; Collins, Michael T; Manning, Elizabeth J B; Naser, Saleh A; Shin, Sung Jae

    2010-02-01

    The etiology of Crohn's disease (CD) is unresolved, but it is likely that an interplay of host genetic factors and environmental triggers is relevant. Mycobacterium paratuberculosis (MAP) has been focused upon as one of these triggers because it causes a similar chronic inflammatory bowel disease in animals. However, the differences among MAP antigens isolated from humans (H-MAP) and cattle (B-MAP) have not been well characterized. In this study, culture filtrate (CF) proteins from MAP isolates were tested with sera from CD patients and healthy controls in enzyme-linked immunosorbent assay (ELISA). Antibody produced by seven CD patients reacted differently according to the antigen source: strong reactivity was seen to H-MAP CF, but not to B-MAP CF. Six proteins, ModD, PepA, transaldolase, EchA9, MAP2120c, and MAP2950c, in H-MAP CF reacting specifically with CD patient sera were identified by liquid chromatography-electrospray ionization-MS. Bioinformatic analysis revealed that ModD and PepA were the same proteins reacting with sera from cattle infected with MAP. The elevated antibody responses of CD patients to rModD and rPepA were confirmed by ELISA (PMAP infections. The study also identified additional proteins potentially useful in the design of assays for human MAP infections. PMID:19878316

  17. Pathology of Paratuberculosis (Johne's Disease) in Clinically Healthy Awassi Sheep with Reference to its Prevalence in Jordan

    Science.gov (United States)

    Paratuberculosis (Johne’s disease) is an incurable infectious, chronically progressive enteric disease affecting domestic and exotic ruminants. The causative agent is Mycobacterium Avium Paratuberculosis. In this study the occurrence of subclinical Johne’s disease in Awassi sheep is investigated. Ou...

  18. Sensibilidad del cultivo de pool fecal para detectar infección por Mycobacterium avium subsp. paratuberculosis en rebaños bovinos de leche y su relación con la prueba de ELISA Sensitivity of pooled faecal culture for detecting Mycobacterium avium subsp. paratuberculosis infection in dairy herds and its relationship with the ELISA test

    Directory of Open Access Journals (Sweden)

    M Pradenas

    2008-01-01

    Full Text Available El objetivo del presente estudio fue evaluar la sensibilidad y especificidad del cultivo de pool fecal como alternativa diagnóstica de paratuberculosis en rebaños lecheros bovinos comparado con el cultivo fecal individual y la prueba de ELISA con la finalidad de reducir costos asociados al diagnóstico de la enfermedad. En 12 rebaños lecheros con antecedentes de paratuberculosis se recolectó un máximo de 50 muestras fecales individuales por rebaño (n=598 y se cultivaron en medio de Herrold en forma individual y en pools estratégicos de 5 y 10 animales. Simultáneamente, de cada animal se recolectó una muestra de sangre para la prueba de ELISA. Se evaluó la sensibilidad y especificidad de las pruebas usando como referencia los cultivos fecales individuales mediante tablas de dos entradas. En 10 (83,3% rebaños y en 42 (7% animales fue posible aislar Map en tanto que el 15,8% y 22% de los pools de 5 y 10 animales, respectivamente, resultaron positivos al cultivo fecal. La sensibilidad de los pools fecales de 5 y 10 animales fue 43,2% y 46,4%, respectivamente, mientras que la sensibilidad del ELISA fue 42,9%. El costo de los pools fecales de 5 animales fue similar al de la prueba de ELISA, pero cuatro veces menor que el costo del cultivo fecal individual; sin embargo, la sensibilidad y especificidad de los pools de 5 animales fue similar a los cultivos individuales. El cultivo de pools fecales demostró ser una buena y costo-efectiva alternativa para el diagnóstico de paratuberculosis para detectar rebaños infectados con MapThe aim of the present study was to evaluate the sensitivity and specificity of pooled faecal culture as a better alternative for the diagnosis of paratuberculosis in dairy herds compared to the conventional faecal culture and the ELISA test. Individual faecal and blood samples were collected from 50 cows in each one of 12 dairy herds (n = 598 with a history of paratuberculosis. Faecal samples were cultured on Herrold

  19. Differential expression of genes encoding CD30L and P-selectin in cattle with Johne's disease: Progress toward a diagnostic gene expression signature

    DEFF Research Database (Denmark)

    Skovgaard, Kerstin; Grell, S. N.; Heegaard, Peter M. H.; Jungersen, Gregers; Pudrith, C. B.; Coussens, P. M.

    Mycobacterium avium subspecies paratuberculosis (Mycobacterium paratuberculosis), the causative agent of paratuberculosis (paraTB) or Johne's disease in ruminants, is a health problem for the global cattle industry with significant economic losses related to decreased milk production and reduced ...

  20. ISOLATION OF THE GENOME SEQUENCE STRAIN MYCOBACTERIUM AVIUM 104 FROM MULTIPLE PATIENTS OVER A 17-YEAR PERIOD

    Science.gov (United States)

    The genome sequence strain 104 of the opportunistic pathogen Mycobacterium avium was isolated form an adult AIDS patient in Southern California in 1983. Isolates of non-paratuberculosis M. avium from 207 other patients in Southern California and elsewhere were examined for genoty...

  1. Analysis of repeated tests for interferon-gamma (IFN-gamma) response and faecal excretion for diagnosis of subclinical paratuberculosis in Danish cattle

    DEFF Research Database (Denmark)

    Huda, A.; Lind, Peter; Christoffersen, Anna-Bodil; Jungersen, Gregers

    A total of 315 cattle were tested for infection with Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) at three consecutive samplings, using the interferon-gamma (IFN-gamma) test on whole blood and bacteriological culture of faecal samples. Of 205 cattle from 10 infected herds 99 ...

  2. Differential cytokine gene expression profiles in the three pathological forms of sheep paratuberculosis

    Directory of Open Access Journals (Sweden)

    Rhind Susan M

    2007-08-01

    Full Text Available Abstract Background Johne's disease is a chronic inflammatory disease of the gut caused by infection with Mycobacterium avium subspecies paratuberculosis (MAP. Symptoms include wasting, diarrhoea, loss of condition and eventual death. Three forms of Johne's disease have been described in sheep – paucibacillary, multibacillary and asymptomatic. The paucibacillary form is characterized by an inflammatory, Th1-type immune response. The multibacillary form of the disease, which disseminates the infection, is characterized by macrophage infiltration mediated by a Th2-type immune response, and asymptomatic animals have no clinical symptoms or pathology but are infected with MAP. What determines these three forms of the disease is unknown. To further understand these differences, we used real-time RT-PCR to compare the expression of thirteen cytokine and cytokine-related genes in ileal tissue from sheep with the three forms of the disease. Results Three pathological forms of sheep paratuberculosis were defined on the basis of histopathology, cytochemistry (Zeihl-Neelsen and IS900 PCR. Paucibacillary lesions have largely T cell and eosinophil infiltration and are ZN negative; multibacillary lesions have macrophage infiltration and large numbers of acid-fast bacteria. The pauci- and multibacillary forms are linked to the differential expression of IFNγ and IL-10 respectively. In addition the increased levels of the proinflammatory cytokines (IL-1β and TNFα, IL-8, IL-18 and TRAF-1 in both diseased forms is indicative of persistent inflammatory lesions. No changes were seen in IL-1α in any sheep ileum tissues. Asymptomatic animals are IS900+ with normal histology but have significantly decreased levels of IL-18 and increased levels TNFα. Conclusion We have quantified the expression levels of thirteen cytokine and cytokine related genes in three forms of ovine paratuberculosis using real-time PCR analyses and confirm that sheep pauci- and

  3. Paratuberculosis control: a review with a focus on vaccination

    OpenAIRE

    Bastida, Felix; Juste, Ramon A.

    2011-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) infection causes in ruminants a regional chronic enteritis that is increasingly being recognized as a significant problem affecting animal health, farming and the food industry due to the high prevalence of the disease and to recent research data strengthening the link between the pathogen and human inflammatory bowel disease (IBD). Control of the infection through hygiene-management measures and test and culling of positive animals has to dat...

  4. Vaccination sequence effects on immunological response and tissue bacterial burden in paratuberculosis infection in a rabbit model.

    Science.gov (United States)

    Arrazuria, Rakel; Molina, Elena; Garrido, Joseba M; Pérez, Valentín; Juste, Ramón A; Elguezabal, Natalia

    2016-01-01

    Paratuberculosis (PTB), a chronic granulomatous enteritis produced by Mycobacterium avium subspecies paratuberculosis (MAP), is considered as one of the diseases with the highest economic impact in the ruminant industry. Vaccination against MAP is recommended during the first months after birth on the basis that protection would be conferred before the first contact with mycobacteria. However, little is known about the therapeutic effect of MAP vaccination in controlled experimental conditions. The current study was designed to evaluate the efficacy of vaccination before and after challenge with MAP in a rabbit infection model. The rabbits were divided into four groups: non-infected control (NIC, n = 4), infected control challenged with MAP (IC, n = 5), vaccinated and challenged 1 month after with MAP (VSI, n = 5) and challenged with MAP and vaccinated 2 months later (IVS, n = 5). The results from this study show a quick increase in IFN-γ release upon stimulation with bovine, avian and johnin PPD in animals vaccinated before MAP challenge. All vaccinated animals show an increased humoral response as seen by western blot and ELISA. The final bacteriology index (considering tissue culture and qPCR) shows that the IC group was the most affected. Vaccination after infection (IVS) produced the lowest bacteriology index showing significant differences with the IC group (p = 0.034). In conclusion, vaccination against MAP shows positive effects in a rabbit model. However, vaccination after infection shows a slightly stronger protective effect compared to vaccination before infection, suggesting a therapeutic effect. This feature could be applied to previously infected adult animals under field conditions. PMID:27496043

  5. Crohn's disease-specific pancreatic autoantibodies are specifically present in ruminants with paratuberculosis: implications for the pathogenesis of the human disease.

    Science.gov (United States)

    Liaskos, Christos; Spyrou, Vassiliki; Roggenbuck, Dirk; Athanasiou, Labrini V; Orfanidou, Timoklia; Mavropoulos, Athanasios; Reinhold, Dirk; Rigopoulou, Eirini I; Amiridis, Georgios S; Billinis, Charalambos; Bogdanos, Dimitrios P

    2013-09-01

    Mycobacterium avium subspecies paratuberculosis (MAP) induces paratuberculosis (ptb) in ruminants and has clinical and histological features resebling Crohn's disease (CD). Pancreatic autoantibodies (PAB) targeting glycoprotein 2 (GP2) are specifically found in CD, but it is currently unknown whether these autoantibodies can be found in ruminants with ptb. IgG anti-MAP and anti-GP2 antibodies were tested by ELISA in 286 ruminants (212 sheep and 74 cattle). PAB testing was performed by indirect immunofluorescence (IIF) using anti-sheep or anti-cattle specific antisera. PCR analysis confirmed the presence of MAP in anti-MAP positive samples. Anti-GP2 antibodies were more prevalent in anti-MAP antibody positive (26.9%) than in anti-MAP negative ruminants (8.7%, p MAP positive sheep compared to 10/142 (7%, p = 0.001) anti-MAP antibody negative and in anti-MAP positive cattle than in negative counterparts (5/8 versus 8/66, p = 0.003). Absorbance values for anti-GP2 antibodies were higher in cattle than in sheep (mean 21 AU/mL ± 25.4SD versus 12.2 AU/mL ± 23 SD, p MAP antibody concentrations. Anti-GP2 antibodies persisted up to 1/1000 and showed the characteristic IIF pancreatic pattern seen by anti-GP2 antibody positive CD samples. This is the first study to demonstrate the presence of CD-specific GP2-reactive pancreatic autoantibodies in MAP-infected ruminants. Our data suggest that CD and ptb are characterised by an antigen-driven loss of immunological tolerance to GP2, implying commonalities in the immunopathogenesis of the human and ruminant inflammatory bowel disorder. PMID:23638886

  6. Mycobacterium Avium Complex (MAC)

    Science.gov (United States)

    ... Select a Language: Fact Sheet 514 Mycobacterium Avium Complex (MAC) WHAT IS MAC? HOW DO I KNOW ... THE BOTTOM LINE WHAT IS MAC? Mycobacterium Avium Complex (MAC) is a serious illness caused by common ...

  7. Invited review: The economic impact and control of paratuberculosis in cattle.

    Science.gov (United States)

    Garcia, A B; Shalloo, L

    2015-08-01

    Paratuberculosis (also called Johne's disease) is a chronic disease caused by Mycobacterium avium ssp. paratuberculosis (MAP) that affects ruminants and other animals. The epidemiology of paratuberculosis is complex and the clinical manifestations and economic impact of the disease in cattle can be variable depending on factors such as herd management, age, infection dose, and disease prevalence, among others. Additionally, considerable challenges are faced in the control of paratuberculosis in cattle, such as the lack of accurate and reliable diagnostic tests. Nevertheless, efforts are directed toward the control of this disease because it can cause substantial economic losses to the cattle industry mainly due to increased premature culling, replacement costs, decreased milk yield, reduced feed conversion efficiency, fertility problems, reduced slaughter values, and increased susceptibility to other diseases or conditions. The variability and uncertainty surrounding the estimations of paratuberculosis prevalence and impact influence the design, implementation, and efficiency of control programs in diverse areas of the world. This review covers important aspects of the economic impact and control of paratuberculosis, including challenges related to disease detection, estimations of the prevalence and economic effects of the disease, and the implementation of control programs. The control of paratuberculosis can improve animal health and welfare, increase productivity, reduce potential market problems, and increase overall business profitability. The benefits that can derive from the control of paratuberculosis need to be communicated to all industry stakeholders to promote the implementation of control programs. Moreover, if the suspected link between Johne's disease in ruminants and Crohn's disease in humans was established, significant economic losses could be expected, particularly for the dairy industry, making the control of this disease a priority across

  8. A Recombinant Multi-Stage Vaccine against Paratuberculosis Significantly Reduces Bacterial Level in Tissues without Interference in Diagnostics

    DEFF Research Database (Denmark)

    Jungersen, Gregers; Thakur, Aneesh; Aagaard, C.; Mikkelsen, H.; Andersen, P.

    -γ assay responses from 40 to 52 weeks compared to non-vaccinated calves. These results indicate the FET11 vaccine can be used to accelerate eradication of paratuberculosis while surveillance or test-and-manage control programs for tuberculosis and Johne’s disease remain in place. Funded by EMIDA ERA......A new (FET11) recombinant vaccine against paratuberculosis was developed based on recombinant antigens from acute and latent stages of Mycobacterium avium subsp. paratuberculosis (Map) infection. In two experiments 28 calves and 15 goats were orally inoculated with live Map in their third week of...... PCR and revealed significantly reduced levels of Map and reduced histopathology. Diagnostic tests for antibody responses and cell-mediated immune responses, used as surrogates of infection, corroborated the observed vaccine efficacy: Five of seven non‐vaccinated calves seroconverted in ID Screen...

  9. Disinfectant Susceptibility of Mycobacterium avium

    OpenAIRE

    Taylor, Robert Henry

    1998-01-01

    Mycobacterium avium, an opportunistic human pathogen, infects between 25 and 50% of advanced-stage acquired immuno-deficiency syndrome (AIDS) patients in the United States. M. avium has been isolated from many environmental sources including: natural waters, soils, and aerosols. M. avium has also been recovered from within municipal and hospital drinking water systems. Rhesus macaques (Macaca mulatta) infected with the simian HIV analog, SIV, have been shown to acquire M. avium infections...

  10. SIMILARIDADES CLINICOPATOLÓGICAS ENTRE PARATUBERCULOSIS Y ENFERMEDAD DE CROHN. ¿POSIBLE VÍNCULO ZOONÓTICO?

    Directory of Open Access Journals (Sweden)

    Johan Calderón

    2008-04-01

    Full Text Available EL artículo presenta a la luz de la literatura científica actual la asociación del Mycobacterium avium subespecie paratuberculosis con la enfermedad de Crohn, las principales similaridades clinicopatológicas. La revisión se limitó a las publicaciones contenidas en el programa HINARI de la WHO, en especial de Elservier Science, Bioline Internacional, Blackwell Publishing, BMJ Publishing, Lippincott Williams & Wilkins, Nature Publishing, PubMed, Springer Science y publicaciones de la Asociación Internacional para estudio de Paratuberculosis. Evidencias significativas apoyan un posible vinculo zoonótico entre las enfermedades. El aislamiento del Mycobacterium avium subespecie paratuberculosis en pacientes con enfermedad de Crohn desde muestras de intestino, leche, sangre periférica y nódulos linfáticos; de igual manera la respuesta inmune especifica a algunos antígenos de Mycobacterium en pacientes con la enfermedad; aunque estos hallazgos tienen cada uno sus propias controversias. La evidencia actualmente disponible no es suficiente para validar o negar al Mycobacterium avium subespecie paratuberculosis como agente causal de por lo menos algunos casos de enfermedad de Crohn. Los estudios son inconclusos en aceptar un vinculo zoonótico dada la naturaleza multifactorial de esta enfermedad. Se requieren estudios para determinar sí Mycobacterium es un agente espectador ó patogénico; paralelamente realizar estudios epidemiológicos a gran escala que permitan analizar la distribución geográfica y temporal de ambas enfermedades.

  11. Histopathological and Serological Studies on Paratuberculosis in Cattle and Buffaloes

    Directory of Open Access Journals (Sweden)

    Arbab Sikandar*, AH Cheema1, M Younus2, A Aslam1, MA Zaman2 and T Rehman3

    2012-10-01

    Full Text Available Paratuberculosis (Johne’s diseases is responsible for massive economic losses to dairy industry, both in the industrially advanced as well as in the developing countries. To detect its occurrence in cattle and buffaloes locally, blood and tissue samples from clinically weak and grossly suspected slaughtered animals were collected from two abattoirs of Jhang, municipal area, Pakistan. Acid-fast smear staining, gross/histopathology and indirect ELISA were done for the detection of Mycobacterium avium subsp. paratuberculosis (MAP. Total 134 samples illustrating gross pathological lesions were collected, only 11.19% (cattle: 6.67%, buffaloes: 12.5% showed acid fast bacilli through smear staining and were taken as confirmed cases. Thickening of intestines alone was not a reliable indicator of Johne’s disease. Tissue sections from intestines and mesenteric lymph nodes from these acid fast positive animals were stained with hematoxylin & eosin (H&E and Ziehl Neelsen (ZN methods. Sum of (15/134 impression smear staining as well as (15/15 tissue sections of the intestines were found ZN positive, and only 6.7% of impression smears and 100% of tissue sections of mesenteric lymph nodes showed acid fast bacilli. Through ELISA, two cattle and five buffaloes (07/134 gave positive optical densities, while one cattle and seven buffaloes (08/134 were judged as doubtful. It is concluded that infection of MAP can be identified by histopathology and ELISA. The present study was the first record of paratuberculosis among the dairy animals slaughtered at Jhang abattoirs. The objective was to compare different methods for the diagnosis of Johne’s disease.

  12. Genetic Association Analysis of Paratuberculosis Forms in Holstein-Friesian Cattle

    Directory of Open Access Journals (Sweden)

    Patricia Vázquez

    2014-01-01

    Full Text Available A genetic susceptibility to Mycobacterium avium subsp. paratuberculosis (MAP infections in ruminants has been longtime suspected to exist. Recently, natural infections in cattle have been reclassified into latent and patent forms based on histopathological findings and their associations with immunological and microbiological variables. This study aims to explore whether these newly defined phenotypes are associated with twenty-four single-nucleotide polymorphisms (SNPs in six bovine candidate genes: nucleotide-binding oligomerization domain 2 (NOD2, solute carrier family 11 member A1 (SLC11A1, nuclear body protein SP110 (SP110, toll-like receptors (TLRs 2 and 4, and CD209 (also known as DC-SIGN, dendritic cell-specific ICAM3-grabbing nonintegrin. SNPs were genotyped for 772 Holstein-Friesian animals (52.6% apparently free; 38.1% latent; 9.3% patent by TaqMan OpenArray technology. Genotypic-phenotypic associations were assessed by logistic regression analysis adjusted for age at slaughter, under five models (codominant, dominant, recessive, overdominant, and log-additive, and corrected for multiple testing. The rs208222804 C allele (CD209 gene was found to be associated with latent paratuberculosis (log-additive model: P<0.0034 after permutation procedure; OR = 0.64, 95% CI = 0.48–0.86. No significant association was detected between any SNP and the patent phenotype. Consequently, CD209 gene may play a key role in the pathogenesis of bovine paratuberculosis.

  13. Probable paratuberculosis in a Sicilian ass.

    Science.gov (United States)

    Dierckins, M S; Sherman, D M; Gendron-Fitzpatrick, A

    1990-02-01

    A presumptive diagnosis of paratuberculosis was made in a Sicilian ass on the basis of a history of chronic diarrhea and weight loss, pasture exposure to a heifer with paratuberculosis confirmed by bacterial culture of feces, postmortem identification of granulomatous inflammation of the intestine containing acid-fast organisms, the absence of acid-fast organisms in extraenteric tissues, and the absence of exposure to tuberculosis. The literature on paratuberculosis in equids is reviewed. The potential for cross-species transmission is emphasized. Justification for consideration of Mycobacterium paratuberculosis infection in the differential diagnoses of equine granulomatous enteritis is discussed. PMID:2298679

  14. Paratuberculose em ruminantes no Brasil Paratuberculosis in ruminants in Brasil: a review

    Directory of Open Access Journals (Sweden)

    Elise M. Yamasaki

    2013-02-01

    Full Text Available A paratuberculose ou doença de Johne é uma enterite granulomatosa causada por Mycobacterium avium subsp. paratuberculosis (Map e comumente afeta ruminantes domésticos, no entanto, pode infectar várias espécies de mamíferos. Está presente nos cinco continentes e é considerada endêmica em algumas regiões pela Organização Internacional de Epizootias (OIE. Pertence à lista de enfermidades notificáveis, que compreende as doenças transmissíveis de importância sócio-econômica e/ou em saúde-pública, cujo controle é necessário para o comércio internacional de animais e alimentos de origem animal. A importância da doença de Johne não se restringe somente aos preju��zos econômicos causados à indústria animal, mas também na possível participação do Map na íleocolite granulomatosa que afeta seres humanos, conhecida como doença de Crohn. No Brasil, a paratuberculose já foi descrita em diversas espécies de ruminantes e em vários estados. Embora os relatos naturais da enfermidade sejam pontuais, acredita-se na possibilidade da transmissão interespecífica e na disseminação do agente através da compra e venda de animais infectados. O objetivo deste artigo foi reunir as informações disponíveis referentes aos aspectos epidemiológicos, clínico-patológicos e laboratoriais da paratuberculose em bovinos, bubalinos, caprinos e ovinos no Brasil, e salientar a necessidade de implementação de medidas de controle sanitário da enfermidade no país, o que possibilitaria a melhoria da qualidade e valorização dos produtos de origem animal no mercado internacional.Paratuberculosis also known as Johne's disease, is a granulomatous enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP, an acid-fast bacillus that preferentially resides within host intestinal macrophages. The condition is most commonly seen in domestic ruminants, however MAP can also infect other mammalian species. Paratuberculosis shows a

  15. Paratuberculosis in sheep and goats.

    Science.gov (United States)

    Windsor, P A

    2015-12-14

    Paratuberculosis is a chronic insidious, often serious, disease of the global small ruminant industries, mainly causing losses from mortalities and reduced productivity on-farm, interference in trading and, in Australia, profound socio-economic impacts that have periodically compromised harmony of rural communities. The pathogenesis, diagnosis, impacts and disease management options for ovine and caprine paratuberculosis are reviewed, comparing current controls in the extensive management system for sheep in wool flocks in Australia with the semi-intensive system of dairy flocks/herds in Greece. Improved understanding of the immune and cellular profiles of sheep with varying paratuberculosis outcomes and the recognition of the need for prolonged vaccination and biosecurity is considered of relevance to future control strategies. Paratuberculosis in goats is also of global distribution although the prevalence, economic impact and strategic control options are less well recognized, possibly due to the relatively meagre resources available for goat industry research. Although there have been some recent advances, more work is required on developing control strategies for goats, particularly in dairy situations where there is an important need for validation of improved diagnostic assays and the recognition of the potential impacts for vaccination. For all species, a research priority remains the identification of tests that can detect latent and subclinical infections to enhance removal of future sources of infectious material from flocks/herds and the food chain, plus predict the likely outcomes of animals exposed to the organism at an early age. Improving national paratuberculosis control programs should also be a priority to manage disease risk from trade. The importance of strong leadership and communication, building trust within rural communities confused by the difficulties in managing this insidious disease, reflects the importance of change management

  16. Whole Genome Association Study of Johne's Disease

    Science.gov (United States)

    Bovine Paratuberculosis, commonly referred to as Johne's disease, is a contagious bacterial disease estimated to be present in over 65% of US dairy herds and results in annual losses in the hundreds of millions of US dollars. Mycobacterium avium subspecies paratuberculosis (MAP) is the bacteria resp...

  17. Activities preceding a decline in the paratuberculosis test prevalence

    DEFF Research Database (Denmark)

    Nielsen, Søren Saxmose; Toft, Nils

    A voluntary control programme on Mycobacterium avium subsp. paratuberculosis (MAP) was initiated in Denmark in 2006 and has since 2007 included 25-28% of the dairy herds and 35-40% of the dairy cows. The programme was complemented with activities aimed to reduce the MAP infection prevalence. A ch...... for a “continuous” flow of diagnostic test information four times annually in each herd. This information can be used for detection and management of infectious animals and for prevalence monitoring....... challenge in evaluation of activities in a national programme it is essentially a sample size of one without a control group. Therefore, the apparent effect of activities on programme level can only be descriptive. Our objective was to describe the decline in the test-prevalence along with the activities...... preceding this decline. The cohort of herds enrolled in 2006-2007 had an average estimated within-herd test-prevalence of 10% at start. By January 2014 this had declined to 2%. The test-prevalence in the cohort of herds enrolled in 2008-2010 started at approximately 6% and by January 2014 was reduced to...

  18. Factors associated with ELISA scores for paratuberculosis in an Angus-Brahman multibreed herd of beef cattle.

    Science.gov (United States)

    Elzo, M A; Rae, D O; Lanhart, S E; Wasdin, J G; Dixon, W P; Jones, J L

    2006-01-01

    Cow and calf genetic and environmental factors were evaluated for their association with ELISA scores for paratuberculosis in a multibreed population of beef cattle. The ELISA scores are a measure of the presence or absence of antibodies against Mycobacterium avium subsp. paratuberculosis in bovine serum. The linear mixed-model analysis used 352 ELISA scores from 238 cows: 51 Angus (A); 34 Brahman (B); 41 (3/4 A 1/4 B); 45 (1/2 A 1/2 B); 34 (1/4 A 3/4 B); and 33 Brangus (5/8 A 3/8 B). Cows were assumed to be unrelated. Year affected (P < 0.001) ELISA scores, but age of cow did not, which was expected to be significant because of the chronic progressive nature of this disease. Important regressions on fixed effects associated with cows were 1) a positive estimate of cow B breed effect (0.59 +/- 0.24; P < 0.017), indicating an upward trend of ELISA scores toward 100% B cows; 2) a negative estimate for weight change from before calving (late November) to the date of the blood sample in May (-0.0062 +/- 0.0019 score/kg; P < 0.002), indicating that poorer maintenance of cow weights was associated with higher ELISA scores; and 3) a positive estimate for days in lactation of cow on the date of the blood sample (0.0086 +/- 0.0034 score/d; P < 0.021), indicating the production of larger amounts of antibodies against Mycobacterium avium subsp. paratuberculosis as lactation progressed. Relevant regressions on fixed effects associated with calves were 1) calf birth weight (-0.022 +/- 0.010 score/kg; P < 0.035), and 2) calf gain from birth to the date of the cow blood sample (-0.0092 +/- 0.0027 score/kg; P < 0.001). These estimates indicate that cows that produced lighter calves at birth and/or calves with slower preweaning growth tended to have greater ELISA scores. Although the sensitivity (percentage of infected animals detected) of ELISA was only 50%, these results suggest that subclinical paratuberculosis may be negatively affecting cows and their offspring. Factors

  19. Resolution of Crohn's disease and complex regional pain syndrome following treatment of paratuberculosis.

    Science.gov (United States)

    Kuenstner, J Todd; Chamberlin, William; Naser, Saleh A; Collins, Michael T; Dow, Coad Thomas; Aitken, John M; Weg, Stuart; Telega, Grzegorz; John, Kuruvilla; Haas, David; Eckstein, Torsten M; Kali, Maher; Welch, Christine; Petrie, Thomas

    2015-04-01

    A cohort of family members with various chronic diseases including Crohn's disease, asthma, complex regional pain syndrome, hypothyroidism, type 1 diabetes mellitus, and lymphangiomatosis and/or evidence of infection by Mycobacterium avium subsp. paratuberculosis (MAP) are described in this series of case reports. MAP was cultured from the blood of three members affected by the first five diseases and there was accompanying elevated anti-MAP IgG in two members. The patient affected by the sixth disease has a markedly elevated anti-MAP titer. The two patients affected by the first four diseases have been treated with a combination of anti-MAP antibiotics and ultraviolet blood irradiation therapy with resolution of the disease symptomatology and inability to culture MAP in post treatment blood samples. These case reports of patients with MAP infections provide supportive evidence of a pathogenic role of MAP in humans. PMID:25852293

  20. Strategies for time of culling in control of paratuberculosis in dairy herds

    DEFF Research Database (Denmark)

    Kudahl, Anne Margrethe Braad; Nielsen, S S; Østergaard, Søren

    2011-01-01

    prices did not affect the ranking between the different culling strategies. Increased market price (20%) of replacement heifers made all culling strategies less profitable and made culling based on a milk yield criterion the most profitable culling strategy for a longer period (11 to 13 yr). A 20......% reduction in heifer price made immediate culling after a positive test the most profitable strategy overall in herds with typical reproduction, and after 9 yr in herd with poor reproduction. To conclude, the ideal culling strategy depends on the aim of intervention, the time horizon, and the reproductive......Effect of time for culling cows infected with Mycobacterium avium ssp. paratuberculosis on prevalence and profitability was identified through simulations. Seven test-and-cull strategies with different culling criteria and no attempts to close infection routes were compared with strategies with (1...

  1. Management of the calving pen is a crucial factor for paratuberculosis control in large dairy herds.

    Science.gov (United States)

    Donat, Karsten; Schmidt, Mandy; Köhler, Heike; Sauter-Louis, Carola

    2016-05-01

    Improvement of hygiene and herd management to reduce the contact of calves with adult cow feces to prevent new infections is one of the basic strategies to manage paratuberculosis-affected dairy herds. Control programs should recommend an evidence-based selection of factors that demonstrably reduce the transmission of the infectious agent and decrease the prevalence of infected cattle to improve acceptance and implementation of the recommended measures among farmers. This study aimed to assess the influence of several management measures on control success in a longitudinal study in 28 large dairy herds with a median size of 415 cows in Thuringia, Germany. The cumulative incidence of cows shedding Mycobacterium avium ssp. paratuberculosis (MAP) per year was determined by individual fecal culture of all cows during 5 consecutive years. Relevant management practices as well as herd size, milk yield, and purchase of cattle were recorded by on-farm risk assessment. Mean holding time of MAP shedders within the herd was calculated from individual data of each shedding cow. Using multiple regression models, separate calving pens for shedders and disinfection of the pen after use were identified as significant risk factors that reduced the cumulative incidence of MAP shedders per year on the herd level. The results provide evidence that, in addition to other factors, calving hygiene and management of the calving pens are crucial for paratuberculosis control, particularly in large dairy herds. Considered together with the outcome from other studies, these results might be important to weight various risk factors and to avoid overburdening and overwhelming farmers and keeping them committed. PMID:26947285

  2. M. paratuberculosis and Parkinson's disease--is this a trigger.

    Science.gov (United States)

    Dow, Coad Thomas

    2014-12-01

    Genetic linkage studies and genome wide analysis have provided insights into complex medical diseases. Mycobacterium avium ss. paratuberculosis (MAP) causes Johne's disease, an important enteric inflammatory disease mostly studied in ruminant animals. MAP is also the putative cause of Crohn's disease. Moreover, MAP has been linked to other inflammatory diseases: sarcoidosis, Blau syndrome, autoimmune diabetes, autoimmune thyroiditis and multiple sclerosis. Genetic studies reveal an association between Parkinson's disease (PD), leprosy and Crohn's disease and since discovered, these findings have been considered "surprising". Autophagy and ubiquitin-proteosome systems are cellular systems that both fight intracellular pathogens (xenophagy) and maintain cellular protein quality control. PD is a common neurodegenerative disease that manifests clinically as a profound movement disorder. The recognized genetic defects of PD create disruption of cellular homeostasis that result in protein folding abnormalities of PD called Lewy bodies. Those same genetic defects are associated with susceptibility to intracellular pathogens, including mycobacteria. It is now understood that PD Lewy body pathology starts in the enteric nervous system and "spreads" to the brain in a retrograde fashion via the vagus nerve. This is the same process by which prions affect the brain. Lewy body pathology of the enteric nervous system predates the Lewy body pathology of the central nervous system (CNS) by years or even decades. This article proposes that genetic defects associated with PD also result in a permissive environment for MAP infection--ineffective xenophagy. It postulates that beginning as an enteric infection, MAP--via the vagus nerve--initiates a pathologic process that results in a targeted neuroinvasion of the CNS. The article proposes that MAP infection and resultant PD pathology are due, in the genetically at-risk and age dependant, to the consumptive exhaustion of the protein

  3. Ocorrência de paratuberculose em búfalos (Bubalus bubalis em Pernambuco Occurrence of paratuberculosis in buffaloes (Bubalus bubalis in Pernambuco

    Directory of Open Access Journals (Sweden)

    Rinaldo A. Mota

    2010-03-01

    Full Text Available A paratuberculose (doença de Johne é uma das doenças de maior importância econômica para ruminantes em vários países e pode representar uma ameaça ao desenvolvimento da pecuária brasileira. É uma doença infecto-contagiosa que provoca enterocolite granulomatosa crônica, incurável e de difícil controle, cujo agente é o Mycobacterium avium subsp. paratuberculosis (MAP. Descreve-se a ocorrência de paratuberculose em um rebanho de búfalos no Estado de Pernambuco, Brasil. Não foi encontrado registro, na literatura, da ocorrência de paratuberculose em búfalos no país. De 100 búfalos, cinco mostravam sinais clínicos característicos da doença. À necropsia de dois animais as lesões estavam restritas ao intestino delgado com evidente espessamento da mucosa, aumento de linfonodos mesentéricos e vasos linfáticos proeminentes e dilatados. À microscopia, observaram-se na mucosa do intestino, infiltrado inflamatório granulomatoso com numerosos macrófagos epitelióides e células gigantes de Langhans, além de bacilos álcool-ácido resistentes (BAAR visualizados através da coloração de Ziehl-Neelsen (ZN. Nos linfonodos mesentéricos, havia espessamento da cápsula e marcada inflamação granulomatosa. O exame direto pela técnica de ZN para pesquisa do bacilo em esfregaços de fezes, raspado de mucosa intestinal e imprint de linfonodos mesentéricos resultou positivo. A PCR IS900 específico de linfonodo mesentérico e mucosa intestinal revelou amplificação de um fragmento de aproximadamente 110pb, confirmada pela comparação com outras sequências de M. avium subsp. paratuberculosis disponíveis no GenBank.Paratuberculosis (PTB is a disease of great economical importance for ruminant in several countries and represents a threat to the development of Brazilian livestock. The contagious disease caused by chronic PTB leads to incurable granulomatous enterocolitis of difficult control. PTB is caused by the Mycobacterium avium

  4. Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

    Science.gov (United States)

    We have previously identified the mycobacterial high G+C codon usage bias as a limiting factor in heterologous expression of MAP proteins from Lb.salivarius, and demonstrated that codon optimisation of a synthetic coding gene greatly enhances MAP protein production. Here, we effectively demonstrate ...

  5. Early weight development of goats experimentally infected with Mycobacterium avium subsp. paratuberculosis.

    Directory of Open Access Journals (Sweden)

    Alyssa N Malone

    Full Text Available Johne's disease is an infectious chronic inflammatory bowel disease in ruminants. The key factor for the management of this disease is an early positive diagnosis. Unfortunately, most diagnostics detect animals with Johne's disease in the clinical stage with positive serology and/or positive fecal cultures. However, for effective management of the disease within herds, it is important to detect infected animals as early as possible. This might only be possible with the help of parameters not specific for Johne's disease but that give an early indication for chronic infections such as weight development. Here we report our findings on the development of total body weight and weight gain during the first six months of goats experimentally infected to induce Johne's disease. Twenty dairy goat kids age 2 to 5 days were included in this study. Goats were divided into two groups: a negative control group and a positive infected group. The weight was obtained weekly throughout the study. Goats of the positive group were infected at the age of seven weeks. We detected significant changes in weight gain and total body weight as early as one week after infection. Differences are significant throughout the six month time period. Weight as a non-specific parameter should be used to monitor infection especially in studies on Johne's disease using the goat model. Our study suggests that goats with Johne's disease have a reduced weight gain and reduced weight when compared with healthy goats of the same age.

  6. Mycobacterium paratuberculosis detection in cow's milk in Argentina by immunomagnetic separation-PCR.

    Science.gov (United States)

    Gilardoni, Liliana Rosa; Fernández, Bárbara; Morsella, Claudia; Mendez, Laura; Jar, Ana María; Paolicchi, Fernando Alberto; Mundo, Silvia Leonor

    2016-01-01

    The aim of this study was to standardize a diagnosis procedure to detect Mycobacterium avium subsp. paratuberculosis (Map) DNA in raw cow milk samples under field conditions. A procedure that combines both immunomagnetic separation and IS900-PCR detection (IMS-IS1 PCR) was employed on milk samples from 265 lactating Holstein cows from Map infected and uninfected herds in Argentina. IMS-IS1 PCR results were analyzed and compared with those obtained from milk and fecal culture and serum ELISA. The extent of agreement between both tests was determined by the Kappa test. IMS-IS1 PCR showed a detection limit of 10(1) CFU of Map/mL of milk, when 50:50 mix of monoclonal and polyclonal antibodies were used to coat magnetic beads. All of the 118 samples from the Map uninfected herds were negative for the set of the tests. In Map infected herds, 80 out of 147 cows tested positive by milk IMS-IS1 PCR (55%), of which 2 (1.4%) were also positive by milk culture, 15 (10%) by fecal culture, and 20 (14%) by serum ELISA. Kappa statistics (95% CI) showed a slight agreement between the different tests (milk of the cows that were not positive in other techniques. This is the first report dealing with the application of IMS-IS1 PCR in the detection of Map in raw milk samples under field conditions in Argentina. PMID:26991290

  7. Two state model for a constant disease hazard in paratuberculosis (and other bovine diseases).

    Science.gov (United States)

    Louzoun, Yoram; Mitchell, Rebecca; Behar, Hilla; Schukken, Ynte

    2015-01-01

    Many diseases are characterized by a long and varying sub-clinical period. Two main mechanisms can explain such periods: a slow progress toward disease or a sudden transition from a healthy state to a disease state induced by internal or external events. We here survey epidemiological features of the amount of bacteria shed during Mycobacterium Avium Paratuberculosis (MAP) infection to test which of these two models, slow progression or sudden transition (or a combination of the two), better explains the transition from intermittent and low shedding to high shedding. Often, but not always, high shedding is associated with the occurrence of clinical signs. In the case of MAP, the clinical signs include diarrhea, low milk production, poor fertility and eventually emaciation and death. We propose a generic model containing bacterial growth, immune control and fluctuations. This proposed generic model can represent the two hypothesized types of transitions in different parameter regimes. The results show that the sudden transition model provides a simpler explanation of the data, but also suffers from some limitations. We discuss the different immunological mechanism that can explain and support the sudden transition model and the interpretation of each term in the studied model. These conclusions are applicable to a wide variety of diseases, and MAP serves as a good test case based on the large scale measurements of single cow longitudinal profiles in this disease. PMID:26092587

  8. Paratuberculosis in sheep: Histochemical, immunohistochemical and in situ hybridization evidence of in utero and milk transmission.

    Science.gov (United States)

    Verin, Ranieri; Perroni, Marco; Rossi, Giacomo; De Grossi, Luigi; Botta, Roberto; De Sanctis, Bruno; Rocca, Stefano; Cubeddu, Tiziana; Crosby-Durrani, Hayley; Taccini, Ennio

    2016-06-01

    To investigate in utero and milk transmission of Mycobacterium avium subsp. paratuberculosis (MAP), tissues from thirteen pregnant sheep, naturally infected and serologically positive to MAP, were examined by means of histochemistry, immunohistochemistry and in situ hybridization. Soon after parturition, ewes were euthanized and tissues samples were collected and prepared. The offspring (18 lambs) were divided into three groups to investigate different routes of MAP transmission. Lambs were sacrificed at three months old and the tissue samples collected, formalin-fixed and paraffin embedded. Hematoxylin and eosin and Ziehl-Neelsen staining methods were performed on fixed tissues for general examination and for detection of acid-fast bacteria. Additionally, immunohistochemical and in situ hybridization techniques were used to detect MAP antigen and MAP DNA respectively. This study of a flock of MAP-infected sheep indicates both in utero and milk transmission of MAP from dams to their offspring. Importantly, this study detected the presence of MAP in the mammary gland and mammary lymph nodes of adult ewes therefore indicating a significant route for the potential exposure to humans from this bacterial infection. PMID:27234557

  9. Evaluation of serum and milk ELISAs for paratuberculosis in Danish dairy cattle

    DEFF Research Database (Denmark)

    Klausen, Joan; Huda, A.; Ekeroth, Lars; Ahrens, Peter

    concurrently from six dairy herds infected with MAP and from two dairy herds without history of infection with MAP. A cut-off value of 7 OD% was used in the ELISAs. At this cut-off value, all six culture-positive herds were positive in the serum ELISA but one was negative in the milk ELISA. All six culture......A milk and a serum ELISA for detection of antibodies against Mycobacterium avium ssp. paratuberculosis (MAP) were evaluated against the complement-fixation test (CFT) and culture of faecal samples from 580 cows collected between August 1996 and December 1996. Milk and serum were obtained......-positive herds were positive in the CFT. In the two culture-negative herds, the serum and the milk ELISA deemed all serum samples negative at this cut-off value, whereas four serum samples from one of these herds were positive in the CFT. The highest cut-off value enabling the milk ELISA to record all six...

  10. Molecular Differentiation of Treponema pallidum Subspecies

    OpenAIRE

    Centurion-Lara, Arturo; Molini, Barbara J.; Godornes, Charmie; Sun, Eileen; Hevner, Karin; Van Voorhis, Wesley C.; Lukehart, Sheila A.

    2006-01-01

    Treponema pallidum includes three subspecies of antigenically highly related treponemes. These organisms cause clinically distinct diseases and cannot be distinguished by any existing test. In this report, genetic signatures are identified in two tpr genes which, in combination with the previously published signature in the 5′ flanking region of the tpp15 gene, can differentiate the T. pallidum subspecies, as well as a simian treponeme.

  11. PARATUBERCULOSIS AND FOOD OF ANIMAL ORIGIN

    OpenAIRE

    E. Sarno; T.A. Sarli

    2013-01-01

    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...

  12. Comparative Functional Genomics and the Bovine Macrophage Response to Strains of the Mycobacterium Genus

    OpenAIRE

    Rue-Albrecht, Kévin; Magee, David A; Killick, Kate E; Nalpas, Nicolas C; Gordon, Stephen V.; MacHugh, David E.

    2014-01-01

    Mycobacterial infections are major causes of morbidity and mortality in cattle and are also potential zoonotic agents with implications for human health. Despite the implementation of comprehensive animal surveillance programs, many mycobacterial diseases have remained recalcitrant to eradication in several industrialized countries. Two major mycobacterial pathogens of cattle are Mycobacterium bovis and Mycobacterium avium subspecies paratuberculosis (MAP), the causative agents of bovine tube...

  13. Serological and molecular diagnosis of paratuberculosis in dairy cattle

    OpenAIRE

    GÜMÜŞSOY, KADİR SEMİH; İÇA, Tuba; ABAY, SEÇİL; AYDIN, Fuat; HIZLISOY, HARUN

    2015-01-01

    In this study, the presence of paratuberculosis was investigated by serological and molecular methods in herds of dairy cattle. Blood, milk, and stool samples of 147 cows aged 2 years old or older with chronic diarrhea were collected. A California mastitis test (CMT) was performed on milk samples. Indirect paratuberculosis enzyme-linked immunosorbent assay (ELISA) test was used for serological investigation. Polymerase chain reaction (PCR) was utilized for molecular identification of Mycobact...

  14. Diagnóstico clínico e histopatológico de paratuberculosis bovina en un hato lechero en Colombia

    Directory of Open Access Journals (Sweden)

    Nicolás Ramírez V.

    2011-12-01

    Full Text Available Objetivo. Analizar retrospectiva y sistemáticamente los hallazgos clínicos e histopatológicos de paratuberculosis bovina Mycobacterium avium subsp. Paratuberculosis (MAP. Los datos fueron obtenidos en diferentes momentos durante un periodo de 8 años (2000-2008 en unhato lechero en Colombia. Materiales y métodos. Se analizó la información documental en 5 casos compatibles con paratuberculisis bovina, así como la información procedente de otros estudios efectuados en el hato sobre la enfermedad realizados paralelamente enel periodo 2000-2008. Resultados. Los 5 animales afectados, presentaron diarrea crónica intermitente, disminución en la producción de leche, enflaquecimiento progresivo, apetito normal, consumo aumentado de agua y constantes fisiológicas normales. A la necropsia se observó engrosamiento de la mucosa intestinal del íleon y de la porción proximal del intestino grueso con múltiples levantamientos y depresiones, que no desaparecían al estirarel tejido. Los vasos sanguíneos mesentéricos se encontraron dilatados y congestivos. Los ganglios linfáticos mesentéricos se encontraron aumentados hasta tres veces, sin clara delimitación de la corteza y de la médula. Las alteraciones histológicas fueron enteritis ylinfadenitis granulomatosa. En tres de los animales se evidenciaron abundantes bacilos ácido alcohol resistentes (BAAR intracelulares en macrófagos, células gigantes y en el intersticio a la coloración de Ziehl-Neelsen. En otros tejidos evaluados no se encontró inflamación de tipo granulomatoso. Conclusiones. Los criterios diagnósticos empleados, así como el análisis de la información diagnóstica generada en otros estudios, permiten confirmar la presencia, circulación y mantenimiento del Mycobacterium avium subsp.paratuberculosis en el hato con un aparente número elevado de animales infectados.

  15. Ovine paratuberculosis: a confirmed case of Johne's disease in Libya.

    Science.gov (United States)

    Sharif, M A M; Farhat, M E; Kraim, E S; Altrabulsi, N A; Kammon, A M; Dayhum, A S; Eldaghayes, I M

    2013-01-01

    Paratuberculosis (Johne's disease) was suspected in a herd of approximately 300 sheep after weight loss and scouring had increased in adult animals despite repeated treatment with anthelmintics, antibiotics, multivitamins and minerals. The herd is located near Tarhouna city. Herd history revealed that a total of 60 ewes showed clinical symptoms and deaths during the last two years. The last case that we attended was submitted to the National Center of Animal Health (NCAH) for a detailed laboratory examination. Gross pathological and histological examination of tissue samples revealed results that were highly comparable with Johne's disease. A definitive diagnosis was made only by histopathological identification of Mycobacterium paratuberculosis in the intestines using Ziehl-Neelsen stain. This is the first documented case of M. paratuberculosis in sheep in Libya. PMID:26623326

  16. Volatile Organic Compound (VOC) Analysis For Disease Detection: Proof Of Principle For Field Studies Detecting Paratuberculosis And Brucellosis

    Science.gov (United States)

    Knobloch, Henri; Köhler, Heike; Nicola, Commander; Reinhold, Petra; Turner, Claire; Chambers, Mark

    2009-05-01

    A proof of concept investigation was performed to demonstrate that two independent infectious diseases of cattle result in different patterns of volatile organic compounds (VOC) in the headspace of serum samples detectable using an electronic nose (e-nose). A total of 117 sera from cattle naturally infected with Mycobacterium avium subsp. paratuberculosis (paraTB, n = 43) or Brucella sp. (n = 26) and sera from corresponding control animals (n = 48) were randomly and analysed blind to infection status using a ST214 e-nose (Scensive Ltd, Leeds, UK). Samples were collected under non-standardised conditions on different farms from the UK (brucellosis) and Germany (paraTB). The e-nose could differentiate the sera from brucellosis infected, paraTB infected and healthy animals at the population level, but the technology used was not suitable for determination of the disease status of individual animals. Nevertheless, the data indicate that there are differences in the sensor responses depending on the disease status, and therefore, it shows the potential of VOC analysis from serum headspace samples for disease detection.

  17. Stochastic models to simulate paratuberculosis in dairy herds

    DEFF Research Database (Denmark)

    Nielsen, Søren Saxmose; Weber, M.F.; Kudahl, Anne Margrethe Braad;

    2011-01-01

    Stochastic simulation models are widely accepted as a means of assessing the impact of changes in daily management and the control of different diseases, such as paratuberculosis, in dairy herds. This paper summarises and discusses the assumptions of four stochastic simulation models and their use...... in the design of certification, surveillance, and control strategies for paratuberculosis in cattle herds. A detailed comparison is made between the Dutch JohneSSim and the Danish PTB-Simherd, using the same context of a set of control strategies in a typical Dutch/Danish herd. The conclusion is that...

  18. 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.

  19. Mycobacterium avium infection improved by microbial substitution of fungal infection

    OpenAIRE

    Yano, Shuichi

    2010-01-01

    We reported a case of Mycobacterium avium infection in which disease activity appeared to have been suppressed after fungal infection. After the increase in β-D-glucan, her symptoms of fever and chest pain disappeared. We think this phenomenon may be microbial substitution and mild fungal infection may improve the activity due to M avium.

  20. 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.

  1. Formulation of indomethacin emulsion using biopolymer of Prunus avium

    OpenAIRE

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

    2012-01-01

    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.

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

    OpenAIRE

    Carter, George; Young, Lowell S.; Bermudez, Luiz E.

    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.

  3. An epidemiological study of paratuberculosis in dairy cattle in Ontario: study design and prevalence estimates.

    OpenAIRE

    NcNab, W B; Meek, A H; Duncan, J R; Martin, S W; Van Dreumel, A A

    1991-01-01

    An observational study involving 304 dairy herds and three abattoirs was conducted between 1986 and 1989 to investigate the epidemiology of paratuberculosis in dairy cattle in Ontario. The objectives of this paper were to describe the method of data collection for the study, to present descriptive production statistics and to present estimates of the prevalence of paratuberculosis among dairy cattle in Ontario. Mycobacterium paratuberculosis was isolated from the distal ileum and/or the ileoc...

  4. The wildlife hosts of Mycobacterium avium subsp. paratuberculosis in the Czech Republic during the years 2002–2007

    Czech Academy of Sciences Publication Activity Database

    Kopečná, M.; Trčka, I.; Lamka, J.; Morávková, M.; Koubek, Petr; Heroldová, Marta; Mrlík, V.; Králová, A.; Pavlík, I.

    2008-01-01

    Roč. 53, č. 8 (2008), s. 420-426. ISSN 0375-8427 R&D Projects: GA ČR GA206/04/2003 Institutional research plan: CEZ:AV0Z60930519 Keywords : Johne’s disease * epidemiology * cattle * non-ruminant species * IS900 PCR Subject RIV: FN - Epidemiology, Contagious Diseases ; Clinical Immunology Impact factor: 0.659, year: 2008 http://journals.uzpi.cz:8050/uniqueFiles/02031.pdf

  5. Dynamics of specific anti-Mycobacterium avium subsp. paratuberculosis antibody response through age

    DEFF Research Database (Denmark)

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

    2013-01-01

    G antibodies on their final test-record were used to estimate age-specific sensitivities (Se). These cows were the infected ones considered to develop disease in a population with a representative age-distribution and were defined as cases. The specificity (Sp) of the test was estimated based on test......-results from 166,905 cows, which had no MAP IgG antibodies in their final four test-records. The Sp, age-specific Se and maximum Se were used to estimate the probability of having HI at a given age resulting in the proportion of infected cows with HI at a given age. For cows 2 years of age, the proportion of...... detectable cases was 0.33, while it was 0.94 for cows 5 years of age. Thus, there was a significant shift in the tip of the iceberg with aging. This study provided a model for estimating the proportion of latent chronic infections that would progress to disease, and the results can be used to model infection...

  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

    result in production of IFN-γ in samples previously exposed to MAP antigens. Whole blood samples were collected from heifers in a Danish dairy herd known to be infected with MAP. The samples were collected on three sample dates, and on each date the blood samples were stimulated with PPDj and recombinant...... time interval from blood sampling to culture. The objective of the study was to assess options for use of day-old blood samples for early-stage diagnosis of MAP infections. Bovine interleukin 12 (IL-12) can induce, and IL-10 reduce, IFN-γ production. Therefore, addition of IL-12 and anti-IL-10 could...... antigens as fresh samples, as day-old samples potentiated with bovine IL-12, and as day-old samples treated with anti-bovine IL-10 antibody. Day-old samples were stored overnight at -4ºC. The correlations between IFN-γ responses in the three types of samples and on different sampling dates were then...

  7. Erratum: Apparent prevalence of beef carcasses contaminated with mycobacterium avium subsp. paratuberculosis sampled from danish slaughter cattle (Veterinary Medicine International)

    DEFF Research Database (Denmark)

    Okura, Hisako; Toft, Nils; Pozzato, Nicola

    2014-01-01

    A tabulation error appeared in Table 1. The fecal contamination data for Age <2 years and >2 years were incorrectly recorded into the table. Whereas the numbers in the table were transposed incorrectly, the actual statistical analyses remain correct.The revised table is reproduced below....

  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, Claus; Nielsen, Søren Saxmose;

    . The study included blood samples from 26 heifers of a MAP infected herd, collected three times with 4 and 5 week interval and blood samples from 60 heifers of a MAP non-infected herd collected once. The IFN-γ responses of the non-infected heifers were used to establish cut-off values for each antigen...

  9. 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

    included blood samples from 26 heifers from a MAP infected herd, collected three times with four to five-week intervals, and blood samples from 60 heifers of a non-infected herd collected once. Heifers of the non-infected herd were used to establish cut-off values for each antigen. The case definition was...

  10. 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;

    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 at...

  11. CD4+ T-cell lines used to evaluate a Mycobacterium avium subsp. paratuberculosis (MAP) peptide vaccine

    DEFF Research Database (Denmark)

    Lybeck, Kari; Sjurseth, Siri K.; Al-Touama, Zainab;

    The aim of the study was to establish a protocol for generation of MAP-specific T-cell lines and to use these lines for evaluation of a peptide vaccine. A protocol for culturing T-cell lines from peripheral blood of goats naturally infected with MAP was established. CD4+ T cells were positively...... selected using an anti CD4 mAb and Dynabeads. Sorted CD4+ cells were cultivated with purified protein derivative from MAP (PPDj) or E. coli sonicate, IL-2, and IL-15. After two cultivation cycles, T cells were tested for recall responses in a proliferative T-cell assay. T-cell line responses were in...... antigens. T-cell lines were now generated by cultivating CD4+ cells with peptides instead of PPDj. Initially, both healthy and MAP-infected goats were vaccinated with 119 peptides defined by in silico analysis. Cellular responses to the peptides were not detected using standard IFN- γ plasma ELISA. However...

  12. CD4+ T-cell lines used to evaluate a Mycobacterium avium subsp. paratuberculosis (MAP) peptide vaccine

    OpenAIRE

    Lybeck, Kari; Sjurseth, Siri K.; Al-Touama, Zainab; Melvang, Heidi Mikkelsen; Aagaard, Claus; Lundegaard, Claus; Jungersen, Gregers; Andersen, Peter; Olsen, Ingrid; Tollefsen, Stig

    2015-01-01

    The aim of the study was to establish a protocol for generation of MAP-specific T-cell lines and to use these lines for evaluation of a peptide vaccine.A protocol for culturing T-cell lines from peripheral blood of goats naturally infected with MAP was established. CD4+ T cells were positively selected using an anti CD4 mAb and Dynabeads. Sorted CD4+ cells were cultivated with purified protein derivative from MAP (PPDj) or E. coli sonicate, IL-2, and IL-15. After two cultivation cycles, T cel...

  13. 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

    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 on...... simulating the effect of different control options at different infection prevalences. The concentration of MAP in milk was estimated using a hierarchical simulation model representing individual cows in a herd while taking both direct and indirect contamination with MAP into account. Parameters included...... true within-herd infection prevalences with cows in different infection stages, which were used to estimate prevalences of milk and fecal shedders, and subsequently MAP concentrations in milk and feces. Cows in different infection stages were considered with different risks of excreting MAP testing...

  14. 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

    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. The...... prevalence of antibody responses of the vaccinated animals was relatively constant from 2 to 6years of age, but decreased in older animals. Among the 140 vaccinated animals 88% tested positive with the IFN-γ test to johnin PPD and 50% responded to PPDb with IFN-γ production above a similar cut-off. Although...... study including 895 vaccinated and 2526 non-vaccinated dairy cows in 9 Danish dairy herds aiming at characterizing the long-term antibody-response to vaccination; and (2) a cross-sectional study of responses in the IFN-γ assay carried out in 140 vaccinated animals in two herds to evaluate the effect of...

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

    OpenAIRE

    Taylor, Robert H.; Joseph O. Falkinham; 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...

  16. Bordetella avium Antibiotic Resistance, Novel Enrichment Culture, and Antigenic Characterization

    OpenAIRE

    Beach, Nathan M; Thompson, Seth; Mutnick, Rachel; Brown, Lisa; Kettig, Gina; Puffenbarger, Robyn; Miyamoto, David; Temple, Louise

    2012-01-01

    Bordetella avium continues to be an economic issue in the turkey industry as the causative agent of bordetellosis, which often leads to serious secondary infections. This study presents a broad characterization of the antibiotic resistance patterns in this diverse collection of B. avium strains collected over the past thirty years. In addition, the plasmid basis for the antibiotic resistance was characterized. The antibiotic resistance pattern allowed the development of a novel enrichment cul...

  17. Hemolysin as a Virulence Factor for Systemic Infection with Isolates of Mycobacterium avium Complex

    OpenAIRE

    Maslow, Joel N.; Dawson, David; Carlin, Elizabeth A.; Holland, Steven M.

    1999-01-01

    Isolates of the Mycobacterium avium complex were examined for hemolysin expression. Only invasive isolates of M. avium were observed to be hemolytic (P < 0.001), with activity the greatest for isolates of serovars 4 and 8. Thus, M. avium hemolysin appears to represent a virulence factor necessary for invasive disease.

  18. Hemolysin as a Virulence Factor for Systemic Infection with Isolates of Mycobacterium avium Complex

    Science.gov (United States)

    Maslow, Joel N.; Dawson, David; Carlin, Elizabeth A.; Holland, Steven M.

    1999-01-01

    Isolates of the Mycobacterium avium complex were examined for hemolysin expression. Only invasive isolates of M. avium were observed to be hemolytic (P < 0.001), with activity the greatest for isolates of serovars 4 and 8. Thus, M. avium hemolysin appears to represent a virulence factor necessary for invasive disease. PMID:9889239

  19. Introgression and dispersal among spotted owl (Strix occidentalis) subspecies

    OpenAIRE

    Funk, W Chris; Forsman, Eric D; Mullins, Thomas D; Haig, Susan M.

    2008-01-01

    Abstract Population genetics plays an increasingly important role in the conservation and management of declining species, particularly for defining taxonomic units. Subspecies are recognized by several conservation organizations and countries and receive legal protection under the US Endangered Species Act (ESA). Two subspecies of spotted owls, northern (Strix occidentalis caurina) and Mexican (S. o. lucida) spotted owls, are ESA-listed as threatened, but the California (S. o. occidentalis) ...

  20. Bordetella avium antibiotic resistance, novel enrichment culture, and antigenic characterization.

    Science.gov (United States)

    Beach, Nathan M; Thompson, Seth; Mutnick, Rachel; Brown, Lisa; Kettig, Gina; Puffenbarger, Robyn; Stockwell, Stephanie B; Miyamoto, David; Temple, Louise

    2012-11-01

    Bordetella avium continues to be an economic issue in the turkey industry as the causative agent of bordetellosis, which often leads to serious secondary infections. This study presents a broad characterization of the antibiotic resistance patterns in this diverse collection of B. avium strains collected over the past thirty years. In addition, the plasmid basis for the antibiotic resistance was characterized. The antibiotic resistance pattern allowed the development of a novel enrichment culture method that was subsequently employed to gather new isolates from diseased turkeys and a healthy sawhet owl. While a healthy turkey flock was shown to seroconvert by four weeks-of-age, attempts to culture B. avium from healthy turkey poults were unsuccessful. Western blot of B. avium strains using pooled serum from diseased and healthy commercial turkey flocks revealed both antigenic similarities and differences between strains. In sum, the work documents the continued exposure of commercial turkey flocks to B. avium and the need for development of an effective, inexpensive vaccine to control spread of the disease. PMID:22721730

  1. 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.

  2. Evaluation of novel oral vaccine candidates and validation of a caprine model of Johne's disease

    OpenAIRE

    Hines, Murray E.; Turnquist, Sue E.; Marcia R. S. Ilha; Rajeev, Sreekumari; Jones, Arthur L.; Whittington, Lisa; Bannantine, John P.; Barletta, Raúl G.; Gröhn, Yrjö T.; Katani, Robab; Talaat, Adel M.; Li, Lingling; Kapur, Vivek

    2014-01-01

    Johne's disease (JD) caused by Mycobacterium avium subspecies paratuberculosis (MAP) is a major threat to the dairy industry and possibly some cases of Crohn's disease in humans. A MAP vaccine that reduced of clinical disease and/or reduced fecal shedding would aid in the control of JD. The objectives of this study were (1) to evaluate the efficacy of 5 attenuated strains of MAP as vaccine candidates compared to a commercial control vaccine using the protocol proposed by the Johne's Disease I...

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

    Directory of Open Access Journals (Sweden)

    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. Johne's disease in cattle is associated with enhanced expression of genes encoding IL-5, GATA-3, tissue inhibitors of matrix metalloproteinases 1 and 2, and factors promoting apoptosis in peripheral blood mononuclear cells

    DEFF Research Database (Denmark)

    Coussens, P.M.; Pudrith, C.B.; Skovgaard, Kerstin; Ren, X.N.; Suchyta, S.P.; Stabel, J.R.; Heegaard, Peter M. H.

    transmission in utero. Once established, infections typically exist in a subclinical state for several years. Recent gene-expression profiling studies suggested the hypothesis that inherent gene-expression profiles in peripheral blood mononuclear cells (PBMCs) front M. paratuberculosis-infected cattle may be......Infection of ruminants with Mycobacterium avium. subspecies paratuberculosis (M. para tuberculosis) leads to a chronic and often fatal granulomatous enteritis known as Johne's disease. Most infections with M. paratuberculosis occur during the first 6 months of life, and there is some evidence for...... different than expression profiles in PBMCs from uninfected controls. If true, this would suggest that it is possible to identify an M. paratuberculosis infection "signature" through transcriptional profiling of peripheral immune cells. In addition, identification of groups or classes of genes showing...

  5. Use of the johnin PPD interferon-gamma assay in control of bovine paratuberculosis

    DEFF Research Database (Denmark)

    Jungersen, Gregers; Mikkelsen, Heidi; Grell, Susanne N.

    2012-01-01

    Although the interferon-gamma (IFN-γ) assay for measurements of cell-mediated immune (CMI) responses to paratuberculosis PPD (johnin) has been available for close to 20 years, the assay has not yet emerged as the long desired test to identify infected animals at an early time point. Among other...... individual animal cannot establish whether the animal is infected or predict the future progression of disease in this animal. The IFN-γ assay should thus be used on a group of animals to test the level of exposure to paratuberculosis bacteria the animals have experienced, and thereby assist in maintaining...... rational in-herd management procedures and in the establishment of paratuberculosis status of a given herd. Indeed, for any diagnostic test applied in paratuberculosis, both the diagnostic target condition and the purpose of the diagnostic testing must be considered before any meaningful estimates of...

  6. Use of the bead beater for preparation of Mycobacterium paratuberculosis template DNA in milk.

    OpenAIRE

    Odumeru, J; Gao, A; Chen, S.; Raymond, M; Mutharia, L

    2001-01-01

    Mycobacterium paratuberculosis is a recognized chronic enteric pathogen that can affect many different species of animals, including primates. It has been suggested that this organism is associated with Crohn's disease in humans, and that milk is a potential source of human exposure to this organism. The limit of the detection of M. paratuberculosis in milk samples by direct PCR was 10(5) cfu/mL if the traditional boiling method was used for template DNA preparation. In this study, an improve...

  7. Bovine paratuberculosis II. A comparison of fecal culture and the antibody response.

    OpenAIRE

    De Lisle, G W; SAMAGH, B.S.; Duncan, J R

    1980-01-01

    Fecal culture for Mycobacterium paratuberculosis and a complement fixing serological test using a carbohydrate antigen were compared for diagnostic efficiency in cattle naturally infected with M. paratuberculosis. Serological reactivity was associated with the persistent fecal shedding of large numbers of bacteria and in the absence of false positives in the population studied, was considered an efficient method for the identification of this segment of infected cattle. Minimally infected cat...

  8. Bovine paratuberculosis I. A herd study using complement fixation and intradermal tests.

    OpenAIRE

    de Lisle, G W; Seguin, P.; SAMAGH, B.S.; Corner, A H; Duncan, J R

    1980-01-01

    A dairy herd (102 cattle) which had been enrolled under a paratuberculosis control program for two years utilizing a complement fixation test (carbohydrate antigen) and intradermal skin test (johnin PPD) was subjected to two further herd tests and followed to slaughter to determine infection status by culture and histology. Mycobacterium paratuberculosis infection was demonstrated in 37 of the animals of which only five were considered reactors on the basis of the last two herd tests applied....

  9. Molecular characterization of Spanish Prunus avium plus trees

    OpenAIRE

    Javier Fernandez-Cruz; Josefa Fernandez-Lopez; Maria Eugenia Miranda-Fontaiña; Raquel Diaz; Gabriel Toval

    2014-01-01

    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....

  10. A new subspecies of Bulbophyllum (Orchidaceae, Bulbophyllinae) from The Society Islands

    OpenAIRE

    Hanna B. Margońska

    2012-01-01

    A new subspecies of the Bulbophyllum tahitense (Orchidaceae), from The Society Islands (French Polynesia) is proposed. A new subspecies is described and illustrated. Keys to taxa of Society Islands genus Bulbophyllum, is added.

  11. Mycobacterium avium subsp. avium distribution studied in a naturally infected hen flock and in the environment by culture, serotyping and IS901 RFLP methods

    OpenAIRE

    2007-01-01

    Mycobacterium avium subsp. avium distribution studied in a naturally infected hen flock and in the environment by culture, serotyping and IS901 RFLP methods CZECH REPUBLIC (Shitaye, J.E.) CZECH REPUBLIC Received: 2006-11-01 Revised: 2007-07-25 Accepted: 2007-07-27

  12. Polyclonal Mycobacterium avium infections in patients with AIDS: variations in antimicrobial susceptibilities of different strains of M. avium isolated from the same patient.

    Science.gov (United States)

    von Reyn, C F; Jacobs, N J; Arbeit, R D; Maslow, J N; Niemczyk, S

    1995-01-01

    Broth microdilution MICs were determined for pairs of strains isolated from five AIDS patients with polyclonal Mycobacterium avium infection. Four (80%) of the five patients were infected simultaneously with strains having different antimicrobial susceptibility patterns. These findings have implications for the interpretation of susceptibility data in M. avium prophylaxis and treatment trials. PMID:7790424

  13. Polyclonal Mycobacterium avium infections in patients with AIDS: variations in antimicrobial susceptibilities of different strains of M. avium isolated from the same patient.

    OpenAIRE

    von Reyn, C F; Jacobs, N J; Arbeit, R D; Maslow, J.N.; Niemczyk, S

    1995-01-01

    Broth microdilution MICs were determined for pairs of strains isolated from five AIDS patients with polyclonal Mycobacterium avium infection. Four (80%) of the five patients were infected simultaneously with strains having different antimicrobial susceptibility patterns. These findings have implications for the interpretation of susceptibility data in M. avium prophylaxis and treatment trials.

  14. An inter-subspecies hybrid rice--Liangyou-Peijiu

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    @@ Liangyou-Peijiu (65002), an inter-subspecies hybrid, was bred in Jiangsu Acad of Agri Sci (JAAS) by two-line method. The female was a low-temperature-sensitive sterile line, Pei'ai 64s (japonica), and the male was 9311.

  15. Psychology Baccalaureates at Work: Major Area Subspecializations, Earnings, and Occupations

    Science.gov (United States)

    Rajecki, D. W.

    2012-01-01

    A Census Bureau national survey identified baccalaureates aged up to 64 years having major area subspecializations labeled "psychology," "industrial/organizational (I/O) psychology," and "counseling psychology." Median 2009 earnings of all such types of baccalaureate psychology alumni were well below the distribution mean of the 153 fields in the…

  16. Monoclonal Antibodies Directed Against the Outer Membrane Protein of Bordetella avium

    OpenAIRE

    Liu, Guanhua; Liang, Manfei; Zuo, Xuemei; Zhao, Xue; Guo, Fanxia; Yang, Shifa; Zhu, Ruiliang

    2013-01-01

    Bordetella avium is the etiologic agent of coryza and rhinotracheitis in poultry. This respiratory disease is responsible for substantial economic losses in the poultry industry. Monoclonal antibodies (MAbs) were produced against the outer membrane proteins (OMPs) of B. avium isolated from diseased chickens. BALB/c mice were immunized with the extracted B. avium OMPs. Then the splenocytes from immunized mice and SP2/0 myeloma cells were fused using PEG 4000. Three stable hybridoma clones (des...

  17. Mycobacterium Avium Arthritis with Extra-articular Abscess in a Patient with Mixed Connective Tissue Disease

    OpenAIRE

    Lee, Choong Won; Sung, Han Dong; Choi, Byong Moon; Kim, Chun Wook; Jun, Su Jin; Min, Sang Jo

    2003-01-01

    A case of Mycobacterium avium arthritis in a 39-year-old female patient with mixed connective tissue disease (MCTD) was reported. An extra-articular abscess had formed outside the knee joint and extended down the calf. A culture was taken of the abscess and synovial fluid disclosed Mycobacteriun avium. This was resistant to most anti-tuberculosis agents. A combination of anti-tuberculosis drugs followed a total resection of the abscess. We concluded that M avium septic arthritis could insidio...

  18. Prevalence of paratuberculosis infection in dairy cattle in Northern Italy

    DEFF Research Database (Denmark)

    Pozzato, N.; Capello, K.; Comin, A.;

    2011-01-01

    estimates are needed for their implementation. Although different herd-level prevalence estimations for paratuberculosis have been reported in Europe, very few studies provided comparable and interpretable values, due to poor study designs and lack of knowledge about the accuracy of the diagnostic tests...... used. To overcome these problems we applied a latent class analysis to the results of two prevalence studies carried out in two neighbouring Northern Italian regions (Lombardy and Veneto) that account for over 50% of the Italian dairy cattle population. Serum samples from a randomly selected number of...... farms in the two regions were analyzed by different ELISA tests. The herd-level Apparent Prevalences (AP) were 48% (190/391) for Lombardy and 65% (272/419) for Veneto. Median within-herd APs were 2.6% and 4.0% for Lombardy and Veneto, respectively. Posterior estimates for the herd-level True Prevalences...

  19. Palynological characteristics of the heterostylous subspecies of Linum mucronatum Bertol

    Directory of Open Access Journals (Sweden)

    Talebi, S. M.

    2014-12-01

    Full Text Available Linum mucronatum is a heterostylous species from sect. Syllinum with four subspecies in Iran. The present study examines palynological characteristics of the heterostylous subspecies of Linum mucronatum, pollen characters of brevistylous individuals (pins as well as longistylous individuals (thrums of these plants by scanning electron microscope and light microscope using the prolonged acetolysis procedure. Sixteen qualitative and quantitative characters were investigated. Pollen equatorial shapes varied between pin and thrum individuals of each subspecies with the exception of L. mucronatum subsp. assyriacum. Pollen sculptures varied between pin and thrum samples of each subspecies and were seen in the gemmate, clavate and baculate shapes. In addition, quantitative palynological characters differed between plants and ANOVA test showed significant variations for traits such as equatorial length, colpi width and apocolpium diameter. Hetrostylous individuals of each subspecies were separated from others in the UPGMA tree and also in the PCO and PCA plots. This study confirmed variations in pollen features between pin and thrum individuals of each subspecies.Linum mucronatum es una especie con heterostilia, que pertenece a la sección Syllinum del género Linum, y tiene cuatro subespecies en Irán. En el presente estudio se examinan las características palinológicas de las subespecies heterostilas de Linum mucronatum Bertol., así como los caracteres polínicos de individuos de los morfos brevistilo (pin y longistilo (thrum de estas plantas, mediante microscopía electrónica de scanning y microscopía óptica usando el método de acetolisis prolongada. Se estudiaron un total de 16 caracteres cualitativos y cuantitativos. La forma ecuatorial del polen varía entre los morfos pin y thrum en todas las subspecies, excepto en L. mucronatum subsp. assyriacum. La ornamentación también varía entre las muestras de morfos pin y thrum de cada subespecie

  20. Mycobacterium paratuberculosis CobT activates dendritic cells via engagement of Toll-like receptor 4 resulting in Th1 cell expansion.

    Science.gov (United States)

    Byun, Eui-Hong; Kim, Woo Sik; Kim, Jong-Seok; Won, Choul-Jae; Choi, Han-Gyu; Kim, Hwa-Jung; Cho, Sang-Nae; Lee, Keehoon; Zhang, Tiejun; Hur, Gang Min; Shin, Sung Jae

    2012-11-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne disease in animals and MAP involvement in human Crohn disease has been recently emphasized. Evidence from M. tuberculosis studies suggests mycobacterial proteins activate dendritic cells (DCs) via Toll-like receptor (TLR) 4, eventually determining the fate of immune responses. Here, we investigated whether MAP CobT contributes to the development of T cell immunity through the activation of DCs. MAP CobT recognizes TLR4, and induces DC maturation and activation via the MyD88 and TRIF signaling cascades, which are followed by MAP kinases and NF-κB. We further found that MAP CobT-treated DCs activated naive T cells, effectively polarized CD4(+) and CD8(+) T cells to secrete IFN-γ and IL-2, but not IL-4 and IL-10, and induced T cell proliferation. These data indicate that MAP CobT contributes to T helper (Th) 1 polarization of the immune response. MAP CobT-treated DCs specifically induced the expansion of CD4(+)/CD8(+)CD44(high)CD62L(low) memory T cells in the mesenteric lymph node of MAP-infected mice in a TLR4-dependent manner. Our results indicate that MAP CobT is a novel DC maturation-inducing antigen that drives Th1 polarized-naive/memory T cell expansion in a TLR4-dependent cascade, suggesting that MAP CobT potentially links innate and adaptive immunity against MAP. PMID:23019321

  1. Cloning and Characterization of CYP51 from Mycobacterium avium

    OpenAIRE

    Pietila, Michael P.; Vohra, Pawan K.; Sanyal, Bharati; Wengenack, Nancy L.; Raghavakaimal, Sreekumar; Thomas, Charles F.

    2006-01-01

    Mycobacterium avium complex (MAC) causes chronic lung disease in immunocompetent people and disseminated infection in patients with AIDS. MAC is intrinsically resistant to many conventional antimycobacterial agents, it develops drug resistance rapidly to macrolide antibiotics, and patients with MAC infection experience frequent relapses or the inability to completely eradicate the infection with current treatment. Treatment regimens are prolonged and complicated by drug toxicity or intoleranc...

  2. Mycobacterium avium complex enteritis in HIV-infected patient

    OpenAIRE

    Masahiro Ishikane; Junko Tanuma

    2014-01-01

    Disseminated Mycobacterium avium complex (MAC) infection is an important AIDS-defining opportunistic infection. The introduction of antimicrobial prophylaxis and antiretroviral therapy (ART) markedly reduced the incidence of disseminated MAC infection and improved the survival of affected individuals. However, it seems that patients with new or recurrent MAC infection are still encountered in clinical practice. Our images captured the characteristic endoscopic findings of MAC duodenitis. The ...

  3. Characterization of IS1245 for Strain Typing of Mycobacterium avium

    Science.gov (United States)

    Pestel-Caron, Martine; Arbeit, Robert D.

    1998-01-01

    IS1245 is an insertion element widely prevalent among isolates of Mycobacterium avium. We used PvuII Southern blots to analyze IS1245 polymorphisms among 159 M. avium isolates (141 clinical isolates from 40 human immunodeficiency virus-infected patients plus 18 epidemiologically related environmental isolates) that represented 40 distinct M. avium strains, as resolved by previous studies by pulsed-field gel electrophoresis (PFGE). All 40 strains carried DNA homologous to IS1245 and thus were typeable. Twenty-five (63%) strains had ≥10 copies of the element, 6 (15%) had 4 to 9 copies, and 9 (23%) had only 1 to 3 copies. Among the last group of nine strains (each of which was distinct by PFGE analysis), IS1245 typing resolved only four patterns and thus provided poor discriminatory power. To evaluate the in vivo stability of IS1245, we analyzed 32 strains for which sets of 2 to 19 epidemiologically related isolates were available. For 19 (59%) of these sets, all isolates representing the same strain had indistinguishable IS1245 patterns. Within eight (25%) sets, one or more isolates had IS1245 patterns that differed by one or two fragments from the modal pattern for the isolates of that strain. Five (16%) sets included isolates whose patterns differed by three or more fragments; on the basis of IS1245 typing those isolates would have been designated distinct strains. IS1245 was stable during in vitro passage, suggesting that the variations observed represented natural translocations of the element. IS1245 provides a useful tool for molecular strain typing of M. avium but may have limitations for analyzing strains with low copy numbers or for resolving extended epidemiologic relationships. PMID:9650925

  4. INFECÇÃO EXPERIMENTAL EM CAPRINOS COM MYCOBACTERIUM PARATUBERCULOSIS DE ORIGEM BOVINA

    Directory of Open Access Journals (Sweden)

    Fernando Padilia Poester

    1994-01-01

    Full Text Available RESUMO Uma amostra de Mycobacterium paratuberculosis de origem bovina foi experimentalmente inoculada por via oral em caprinos. Após três anos de observação, os autores obtiveram a reprodução da infecção em dois dos três caprinos inoculados, concluindo que os caprinos são suscetíveis a amostras bovinas de M. paratuberculosis. A possibilidade de transmissão intra-uterina da bactéria, embora tentada não foi possível de ser demonstrada.

  5. 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...

  6. Comparative Serological and Random Amplified Polymorphic DNA Typing for Bordetella avium Isolates in China

    Directory of Open Access Journals (Sweden)

    Ping-Ping Yang, Rong-De Ma1, Xue Zhao and Rui-Liang Zhu*

    2012-10-01

    Full Text Available To study the similarity among Bordetella avium isolates in China, antigens and diagnostic antiserum of 22 B. avium isolates were prepared for serotyping, and a set of 20 commercially available primers was screened out to identify suitable primers for random amplified polymorphic DNA fingerprinting (RAPD analysis in this study. Twenty-two B. avium isolates were divided into two serovars (A and B based on their reaction in the plate-agglutination test. Four primers R1, R2, R4 and R10 resulted in informative fingerprints and were used to evaluate the B. avium isolates. Based on their RAPD patterns, a dendrogram allowed the separation of the B. avium isolates into six genetic similarity clusters. However, no direct correlation was observed between serotypes and RAPD typing among the isolates.

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

    Directory of Open Access Journals (Sweden)

    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.

  8. Seroprevalence of infectious diseases in saiga antelope (Saiga tatarica tatarica) in Kazakhstan 2012-2014.

    Science.gov (United States)

    Orynbayev, Mukhit B; Beauvais, Wendy; Sansyzbay, Abylay R; Rystaeva, Rashida A; Sultankulova, Kulyaisan T; Kerimbaev, Aslan A; Kospanova, Madina N; Kock, Richard A

    2016-05-01

    286 serum samples were collected from three sub-populations of saiga in Kazakhstan (Betpakdala, Ustyurt and Volga-Ural) between 2012 and 2014, and were tested for the presence of antibodies to Brucella spp., bluetongue virus, peste des petits ruminants (PPR) virus, Akabane virus, Schmallenberg virus, Chlamydophila, Toxoplasma, Mycobacterium avium subspecies paratuberculosis and Coxiella burnetii (Q Fever). Seropositives to Coxiella burnetii of saiga were detected and the adjusted seroprevalence of Q Fever antibodies was 0.07 (95% confidence interval (CI): 0.03-0.10). Seropositives to Akabane virus were detected in all three populations and the adjusted seroprevalence values for this virus were very high (all were>0.13). Lower adjusted seroprevalence values were estimated for PPR Virus and Mycobacterium avium subsp. paratuberculosis (0.005 and 0.006). No seropositives for bluetongue, Toxoplasma, Brucella or Schmallenberg were detected. PMID:27094147

  9. Comparison of four different culture media for growth of Mycobacterium avium subsp. avium isolated from naturally infected lofts of domestic pigeons.

    Directory of Open Access Journals (Sweden)

    Mansour Mayahi

    2013-12-01

    Full Text Available Diagnosis of avian tuberculosis by conventional culture method is still considered as the "gold standard" technique. The main objective of this study was to compare growth of Mycobacterium avium subsp. avium on four specific Mycobacterial cultures such as glycerinated Lowenstein-Jensen medium, pyruvate-enriched Lowenstein-Jensen medium, mycobactin J-supplemented Herrold-egg yolk medium and plain Herrold-egg yolk medium.Eighty out of more than 600 pigeons were selected based on their clinical signs and poor health conditions. The birds were numbered and their clinical signs were registered in the working sheets, and under standard condi-tion, euthanized, subjecting to necropsy examinations, followed by bacterial culture on four specific media for Mycobacterium avium subsp. avium, including glycerinated Lowenstein-Jensen (LJG medium, pyruvate-enriched Lowenstein-Jensen medium (LJP, mycobactin J-supplemented Herrold-egg yolk medium and plain Herrold-egg yolk medium.Fifty one Mycobacterium avium subsp. avium were isolated from pigeons. Mycobactin J-supplemented Herrold-egg yolk media yielded greater number of colonies in shorter incubation time in compare with other media.It was concluded that most of the isolates need mycobactin as a growth factor.

  10. Differentiation of Melipona quadrifasciata L. (Hymenoptera, Apidae, Meliponini) subspecies using cytochrome b PCR-RFLP patterns

    OpenAIRE

    Rogério O. Souza; Geraldo Moretto; Arias, Maria C.; Marco A. Del Lama

    2008-01-01

    Melipona quadrifasciata quadrifasciata and M. quadrifasciata anthidioides are subspecies of M. quadrifasciata, a stingless bee species common in coastal Brazil. These subspecies are discriminated by the yellow stripe pattern of the abdominal tergites. We found Vsp I restriction patterns in the cytochrome b region closely associated to each subspecies in 155 M. quadrifasciata colonies of different geographical origin. This mitochondrial DNA molecular marker facilitates diagnosis of M. quadrifa...

  11. Molecular variation of Trypanosoma brucei subspecies as revealed by AFLP fingerprinting

    OpenAIRE

    Agbo, E.E.C.; Majiwa, P.A.O.; Claassen, H.J.H.M.; Pas, te, M.F.W.

    2002-01-01

    Genetic analysis of Trypanosoma spp. depends on the detection of variation between strains. We have used the amplified fragment length polymorphism (AFLP) technique to develop a convenient and reliable method for genetic characterization of Trypanosome (sub)species. AFLP accesses multiple independent sites within the genome and would allow a better definition of the relatedness of different Trypanosome (sub)species. Nine isolates (3 from each T. brucei subspecies) were tested with 40 AFLP pri...

  12. The effect of paratuberculosis on milk yield--A systematic review and meta-analysis.

    Science.gov (United States)

    McAloon, Conor G; Whyte, Paul; More, Simon J; Green, Martin J; O'Grady, Luke; Garcia, AnaBelen; Doherty, Michael L

    2016-02-01

    Bovine paratuberculosis is a disease characterized by chronic granulomatous enteritis causing protein-losing enteropathy. Adverse effects on animal productivity are key drivers in the attempt to control paratuberculosis at the farm level. Economic models require an accurate estimation of the production effects associated with paratuberculosis. The aim of this study was to conduct a systematic review and meta-analysis to investigate the effect of paratuberculosis on milk production. A total of 20 effect estimates from 15 studies were included in the final meta-analysis. Substantial between-study heterogeneity was observed. Subgroup analysis by case definition and study design was carried out to investigate heterogeneity. The majority of between-study variation was attributed to studies that defined cases on serology. Calculation of a pooled effect estimate was only appropriate for studies that defined cases by organism detection. A reduction in milk yield, corrected for lactation number and herd of origin of 1.87 kg/d, equivalent to 5.9% of yield, was associated with fecal culture or PCR positivity in individual cows. PMID:26686704

  13. Differentiation of Melipona quadrifasciata L. (Hymenoptera, Apidae, Meliponini subspecies using cytochrome b PCR-RFLP patterns

    Directory of Open Access Journals (Sweden)

    Rogério O. Souza

    2008-01-01

    Full Text Available Melipona quadrifasciata quadrifasciata and M. quadrifasciata anthidioides are subspecies of M. quadrifasciata, a stingless bee species common in coastal Brazil. These subspecies are discriminated by the yellow stripe pattern of the abdominal tergites. We found Vsp I restriction patterns in the cytochrome b region closely associated to each subspecies in 155 M. quadrifasciata colonies of different geographical origin. This mitochondrial DNA molecular marker facilitates diagnosis of M. quadrifasciata subspecies matrilines and can be used to establish their natural distribution and identify hybrid colonies.

  14. The complete mitochondrial genome of a Chinese rufous horseshoe bat subspecies, Rhinolophus sinicus sinicus (Chiroptera: Rhinolophidae).

    Science.gov (United States)

    Sun, Haijian; Dong, Ji; Shi, Huizhen; Ren, Min; Hua, Panyu

    2016-09-01

    There are two subspecies of Rhinolophus sinicus currently recognized in China. In this study, using next generation sequencing approaches, the complete mitochondrial genome of one subspecies R. s. sinicus was obtained. The total length of the genome sequence is 16,898 bp. The arrangement and contents of R. s. sinicus mitochondrial genes exhibit high similarity with other bats of family Rhinolophida. Phylogenetic reconstructions support the sister relationship of the two subspecies and confirm the subspecies status of our specimen. PMID:25740216

  15. Interference of paratuberculosis with the diagnosis of tuberculosis in a goat flock with a natural mixed infection

    OpenAIRE

    Álvarez, Julio; De Juan, Lucía; Bezos, Javier; Romero, Beatriz; Sáez, Jose Luis; Gordejo, F.J. Reviriego; Briones, Víctor; Moreno, Miguel Ángel; Mateos, Ana; Domínguez, Lucas; Aranaz, Alicia

    2008-01-01

    Interference of paratuberculosis with the diagnosis of tuberculosis in a goat flock with a natural mixed infection SPAIN (Alvarez, Julio) SPAIN Received: 2007-07-06 Revised: 2007-08-14 Accepted: 2007-08-17

  16. [Treponema pallidum subspecies pallidum -- the causative agent of neurosyphilis].

    Science.gov (United States)

    Salavec, Miloslav; Boštíková, Vanda; Vaňásková, Zuzana; Smetana, Jan; Sleha, Radek; Coufalová, Monika; Plíšek, Stanislav; Špliňo, Miroslav; Štěpánová, Vlasta; Boštík, Pavel

    2013-09-01

    Neurosyphilis is defined as infection of central nervous system by Treponema pallidum subspecies pallidum. Neurosyphilis can develop at any stage after initial infec-tion and is reflected in laboratory results. The pathogenesis of neurosyphilis is similar to that of classical form of syphilis. Individuals with persistent abnormalities in the cerebrospinal fluid are at risk of the development of clinical manifestations. Proper understanding of particular forms of neurosyphilis for differential diagnosis is important to determine potential risk of the development of progressive disease in neurology. PMID:24116696

  17. Serovars of Mycobacterium avium Complex isolated from patients in Denmark

    DEFF Research Database (Denmark)

    Askgaard, D. S.; Giese, Steen Bjørck; Thybo, S.; Lerche, A.; Bennedsen, J.

    1994-01-01

    Danish isolates of Mycobacterium avium complex were serotyped by the use of seroagglutination. The most prevalent serovars among patients with AIDS (n = 89) were 4 and 6, while among non-AIDS patients the most prevalent serovars were 1, 6, and 4, with no major differences between those in patients...... with pulmonary disease (n = 65) and those in patients with lymph node infection (n = 58). The results suggest a Scandinavian distribution of serovars with a predominance of serovar 6 and fail to demonstrate any selective protection against different serovars by Mycobacterium bovis ECG vaccination....

  18. Subspecies in Przewalski's gazelle Procapra przewalskii and its conservation implication

    Institute of Scientific and Technical Information of China (English)

    MARDAN Turghan; JIANG ZhiGang; GROVES Colin P; YANG Ji; FANG HongXia

    2013-01-01

    Przewalski's gazelle Procapra przewalskii is an endangered species endemic to China.A question remains about subspecific variation in this species.Skulls of Przewalski's gazelle collected from its current remnant ranges around the Qinghai Lake in combination with those collected prior to the 20th century were measured and analyzed using Hierarchical Cluster Analysis in order to clarify the question.Unexpectedly,P.p.diversicornis,extirpated from its historic range,has spread to the Qinghai Lake region where it has replaced nominotypical P.p.przewalskii and is now restricted to a few small isolated populations around the lake.We discuss the causes of this unexpected replacement.In this study,we discuss the possibility of a new form,possibly a new subspecies,in the Guide Basin,adjacent to Qinghai Lake; it is unclear whether the new form has long existed and was only discovered in recent years,or whether it evolved in recent times due to the geographical isolation and anthropogenic landscape features.The study sheds light on the processes of microevolution and subspeciation in Procapra przewalskii,and based on the findings,we propose measures for conservation strategies for Przewalski's gazelle.

  19. 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.

  20. 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

    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...

  1. Intracellular pH of Mycobacterium avium subsp. paratuberculosis following exposure to antimicrobial compounds monitored at the single cell level

    DEFF Research Database (Denmark)

    Gaggìa, Francesca; Nielsen, Dennis Sandris; Biavati, Bruno;

    2010-01-01

    H(i)) of one strain of MAP after exposure to nisin and neutralized cell-free supernatants (NCSs) from five bacteriocin-producing lactic acid bacteria (LAB) with known probiotic properties. The evaluation of pH(i) by FRIM provides information about the physiological state of bacterial cells, bypassing the...... long and problematic incubations needed for methods relying upon growth of MAP such as determination of colony forming units. The FRIM results showed that both nisin and the cell-free supernatant from Lactobacillus plantarum PCA 236 affected the pH(i) of MAP within a few hours. However, monitoring the...

  2. 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; Jungersen, Gregers

    2012-01-01

    of the study were to evaluate immunogenicity and specificity of 14 novel recombinant antigens for use in the IFN-¿ assay and to assess the consistency of IFN-¿ responses with repeated samplings. The antigens used were 4 ESAT-6 family members, 4 latency proteins, 4 secreted proteins including Ag85B, 3...... were positive to PPDj were non-cases and one case was negative to PPDj but positive to all other tested antigens. Furthermore, PPDj produced elevated IFN-¿ responses in both the infected and non-infected herds and showed low consistency. Immunogenicity was highest for the group of latency proteins (0...

  3. 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

    in farm bulk tank milk and simulating the effect of direct and indirect contamination with MAP. The effect of discarding milk from test-positive cows at different prevalences was assessed. The concentration of MAP in milk was estimated using a simulation model, while taking direct and indirect...... 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 was...

  4. Dam's infection progress and within-herd prevalence as predictors of Mycobacterium avium subsp. paratuberculosis ELISA response in Danish Holstein cattle

    DEFF Research Database (Denmark)

    Nielsen, Søren Saxmose; Hansen, Kira Frello; Kvist, Louise;

    2016-01-01

    correct for the effect of age. The results showed that both the within-herd test-prevalence and the dam's infection progress were significant predictors, while the dams that had tested positive when giving birth and up to 0.7 years after were more likely to have offspring that would test positive. The...... testing positive, compared to animals born in a low prevalence herd. The variance heterogeneity reduced 81% through the included predictors. The results of this study suggest that irrespective of the prevalence, offspring of dams with MAP specific antibodies should be considered as high-risk animals when...... odds of testing positive were about 1.5 to 2.5 times higher for these offspring, compared to offspring of dams that never tested positive. Furthermore, offspring born in high (>5% ELISA-positive) and medium (2.5 to 5% ELISA-positive) prevalence herds had 9 and 3, respectively, times higher odds of...

  5. Evaluation of critical control points in dairy herd management to reduce transmission of Mycobacterium avium subsp. paratuberculosis - results from controlled clinical trials

    Science.gov (United States)

    Johne's disease is a chronic, debilitating intestinal disorder in cattle, sheep and wild ruminants, characterized by diarrhea, weight loss and death. Animals usually become infected when they are young by ingesting feces or milk containing the causative bacteria. However, clinical signs of disease...

  6. 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...

  7. Floral scent variation in two Antirrhinum majus subspecies influences the choice of naïve bumblebees

    OpenAIRE

    Dormont, Laurent; Schatz, Bertrand; Giurfa, Martin; Simon, Valérie; Raynaud, Christine; Chave, Jérôme

    2011-01-01

    Two wild subspecies of snapdragon, Antirrhinum majus, subspecies pseudomajus and striatum, differ in floral color and can be visually discriminated by insect visitors. The extent to which olfactory cues derived from floral scents contribute to discrimination between snapdragon subspecies is however unknown. We tested whether these two subspecies differ in floral scent and whether these olfactory differences are used by bumblebees (Bombus terrestris) to discriminate between them. We grew indiv...

  8. Differentiation of subspecies and sexes of Beringian Dunlins using morphometric measures

    Science.gov (United States)

    Gates, H. River; Yezerinac, Stephen; Powell, Abby N.; Tomkovich, Pavel S.; Valchuk, Olga P.; Lanctot, Richard B.

    2013-01-01

    Five subspecies of Dunlins (Calidris alpina) that breed in Beringia are potentially sympatric during the non-breeding season. Studying their ecology during this period requires techniques to distinguish individuals by subspecies. Our objectives were to determine (1) if five morphometric measures (body mass, culmen, head, tarsus, and wing chord) differed between sexes and among subspecies (C. a. actites, arcticola, kistchinski, pacifica, and sakhalina), and (2) if these differences were sufficient to allow for correct classification of individuals using equations derived from discriminant function analyses. We conducted analyses using morphometric data from 10 Dunlin populations breeding in northern Russia and Alaska, USA. Univariate tests revealed significant differences between sexes in most morphometric traits of all subspecies, and discriminant function equations predicted the sex of individuals with an accuracy of 83–100% for each subspecies. We provide equations to determine sex and subspecies of individuals in mixed subspecies groups, including the (1) Western Alaska group of arcticola and pacifica (known to stage together in western Alaska) and (2) East Asia group of arcticola, actites, kistchinski, and sakhalina (known to winter together in East Asia). Equations that predict the sex of individuals in mixed groups had classification accuracies between 75% and 87%, yielding reliable classification equations. We also provide equations that predict the subspecies of individuals with an accuracy of 22–96% for different mixed subspecies groups. When the sex of individuals can be predetermined, the accuracy of these equations is increased substantially. Investigators are cautioned to consider limitations due to age and feather wear when using these equations during the non-breeding season. These equations will allow determination of sexual and subspecies segregation in non-breeding areas, allowing implementation of taxonomic-specific conservation actions.

  9. Formulation and efficacy of liposome-encapsulated antibiotics for therapy of intracellular Mycobacterium avium infection.

    OpenAIRE

    Oh, Y. K.; Nix, D E; Straubinger, R M

    1995-01-01

    Mycobacterium avium is an intracellular pathogen that can invade and multiply within macrophages of the reticuloendothelial system. Current therapy is not highly effective. Particulate drug carriers that are targeted to the reticuloendothelial system may provide a means to deliver antibiotics more efficiently to M. avium-infected cells. We investigated the formulation of the antibiotics ciprofloxacin and azithromycin in liposomes and tested their antibacterial activities in vitro against M. a...

  10. Effect of growth in biofilms upon antibiotic and chlorine susceptibility of Mycobacterium avium and Mycobacterium intracellulare

    OpenAIRE

    Steed, Keesha

    2003-01-01

    ABSTRACT Mycobacterium avium and Mycobacterium intracellulare are environmental opportunistic pathogens whose source for human infection is water and soil. M. avium and M. intracellulare cause pulmonary infections (tuberculosis) in immunocompetent individuals and bacteremia in immunodeficient individuals (e.g. AIDS). One factor likely influencing the lack of success of antibiotic therapy in patients would be their ability to form biofilms. Growth in biofilms might result in antimicrob...

  11. A new insight in desiccation tolerance and cryopreservation of mazzard cherry (Prunus avium L.) seeds

    OpenAIRE

    Michalak Marcin; Plitta-Michalak Beata P.; Chmielarz Paweł

    2015-01-01

    A variable response of Prunus avium L. seeds to desiccation and storage in liquid nitrogen (LN) has been reported in the literature. The majority of these experiments were conducted on initially dried seeds. The desiccation and LN exposure tolerance of fresh P. avium seeds is unknown. In the present study, fresh seeds were used to determine seed response to desiccation and cryopreservation. Desiccation of seeds from a moisture content (MC) of 19.7-20.2% to 10.1-10.9% o...

  12. De novo assembly of sweet cherry (Prunus avium) transcriptome from 454 sequencing data

    OpenAIRE

    Le Dantec, Loick; Quero-Garcia, José; Dirlewanger, Elisabeth

    2012-01-01

    Very few genomic resources are actually available for sweet cherry. Emergence and generalization of next‐generation DNA sequencing technologies that reduce cost, labor, and time, provide the opportunity to conduct large‐scale genomic projects at lower cost even for non‐model organisms like prunus avium. In order to obtain a panel of prunus avium expressed genes for functional studies and candidate gene strategy, a normalized complementary DNA library has been sequenced using high throughput 4...

  13. Molecular Analysis of Mycobacterium avium Isolates by Using Pulsed-Field Gel Electrophoresis and PCR

    OpenAIRE

    Pestel-Caron, Martine; Graff, Gabriel; Berthelot, Gilles; Pons, Jean-Louis; Lemeland, Jean-François

    1999-01-01

    Genetic relationships among 46 isolates of Mycobacterium avium recovered from 37 patients in a 2,500-bed hospital from 1993 to 1998 were assessed by pulsed-field gel electrophoresis (PFGE) and PCR amplification of genomic sequences located between the repetitive elements IS1245 and IS1311. Each technique enabled the identification of 27 to 32 different patterns among the 46 isolates, confirming that the genetic heterogeneity of M. avium strains is high in a given community. Furthermore, this ...

  14. Characterization of the Fucosylation Pathway in the Biosynthesis of Glycopeptidolipids from Mycobacterium avium Complex▿

    OpenAIRE

    Miyamoto, Yuji; Mukai, Tetsu; Maeda, Yumi; Nakata, Noboru; Kai, Masanori; Naka, Takashi; Yano, Ikuya; Makino, Masahiko

    2007-01-01

    The cell envelopes of several species of nontuberculous mycobacteria, including the Mycobacterium avium complex, contain glycopeptidolipids (GPLs) as major glycolipid components. GPLs are highly antigenic surface molecules, and their variant oligosaccharides define each serotype of the M. avium complex. In the oligosaccharide portion of GPLs, the fucose residue is one of the major sugar moieties, but its biosynthesis remains unclear. To elucidate it, we focused on the 5.0-kb chromosomal regio...

  15. Mycobacterium avium serovars 2 and 8 infections elicit unique activation of the host macrophage immune responses.

    Science.gov (United States)

    Cebula, B R; Rocco, J M; Maslow, J N; Irani, V R

    2012-12-01

    Mycobacterium avium is an opportunistic pathogen whose pathogenesis is attributed to its serovar-specific glycopeptidolipid (ssGPL), which varies among its 31 serovars. To determine if the presence and type of ssGPLs contribute to M. avium pathogenesis, we infected murine macrophages (mφs) with two M. avium wild type (wt) serovars (2 and 8) and their serovar-null strains. We examined the influence of ssGPL (presence and type) on cytokine production in non-activated (-IFN-γ) and activated (+IFN-γ) mφs, and the bacterial intra-mφ survival over a 6-day infection process. Serovar-2 infections activated TNF-α production that increased over the 6 day period and was capable of controlling the intra-mφ serovar-2 null strain. In contrast, the serovar-8 infection stimulated a strong pro-inflammatory response, but was incapable of removing the invading pathogen, maybe through IL-10 production. It was clear that the intracellular growth of serovar-null in contrast to the wt M. avium strains was easily controlled. Based on our findings and the undisputed fact that M. avium ssGPL is key to its pathogenesis, we conclude that it is not appropriate to dissect the pathogenesis of one M. avium serovar and apply those findings to other serovars. PMID:22991047

  16. Curcurbita pepo subspecies delineates striped cucumber beetle (Acalymma vittatum) preference

    Science.gov (United States)

    Brzozowski, L; Leckie, B M; Gardner, J; Hoffmann, M P; Mazourek, M

    2016-01-01

    The striped cucumber beetle (Acalymma vittatum (F.)) is a destructive pest of cucurbit crops, and management could be improved by host plant resistance, especially in organic farming systems. However, despite the variation in striped cucumber beetle preference observed within the economically important species, Cucurbita pepo L., plant breeders and entomologists lacked a simple framework to classify and exploit these differences. This study used recent phylogenetic evidence and bioassays to organize striped cucumber beetle preference within C. pepo. Our results indicate preference contrasts between the two agriculturally relevant subspecies: C. pepo subsp. texana and C. pepo subsp. pepo. Plants of C. pepo subsp. pepo were more strongly preferred than C. pepo subsp. texana plants. This structure of beetle preference in C. pepo will allow plant breeders and entomologists to better focus research efforts on host plant non-preference to control striped cucumber beetles. PMID:27347423

  17. Economy, efficacy, and feasibility of a risk-based control program against paratuberculosis

    DEFF Research Database (Denmark)

    Kudahl, Anne Braad; Nielsen, Søren Saxmose; Østergaard, Søren

    2008-01-01

    Long-term effects of paratuberculosis on within-herd prevalence and on-farm economy of implementing risk-based control strategies were compared with alternative strategies by using a herd-simulation model. Closing transmission routes is essential for effective control of paratuberculosis. However......, many farmers lack the resources to carry out these procedures for all cows in the herd. When using risk-based control strategies 1) all cows are tested quarterly with a milk ELISA, 2) specific cows with a high risk of being infectious are identified, and 3) the farmer can focus only on these infectious...... animals to close infection routes. In this way the workload can be reduced, making these control strategies more feasible. This study evaluates potential long-term effects of the risk-based approach compared with non-risk-based strategies by simulations conducted with the herd-simulation model PTB...

  18. Consensus-based reporting standards for diagnostic test accuracy studies for paratuberculosis in ruminants

    DEFF Research Database (Denmark)

    Gardner, Ian A.; Nielsen, Søren Saxmose; Whittington, Richard;

    2011-01-01

    studies such as herd tests, potential use of experimental challenge studies, a more diverse group of testing purposes and sampling designs, and the widespread lack of an ante-mortem reference standard with high sensitivity and specificity. The objective of the present study was to develop a modified...... Reporting of Animal Diagnostic Accuracy Studies for paratuberculosis), should facilitate improved quality of reporting of the design, conduct and results of paratuberculosis test accuracy studies which were identified as “poor” in a review published in 2008 in Veterinary Microbiology......The Standards for Reporting of Diagnostic Accuracy (STARD) statement (www.stard-statement.org) was developed to encourage complete and transparent reporting of key elements of test accuracy studies in human medicine. The statement was motivated by widespread evidence of bias in test accuracy...

  19. Use of the bead beater for preparation of Mycobacterium paratuberculosis template DNA in milk.

    Science.gov (United States)

    Odumeru, J; Gao, A; Chen, S; Raymond, M; Mutharia, L

    2001-10-01

    Mycobacterium paratuberculosis is a recognized chronic enteric pathogen that can affect many different species of animals, including primates. It has been suggested that this organism is associated with Crohn's disease in humans, and that milk is a potential source of human exposure to this organism. The limit of the detection of M. paratuberculosis in milk samples by direct PCR was 10(5) cfu/mL if the traditional boiling method was used for template DNA preparation. In this study, an improved method for template DNA preparation was examined. The method involves the use of a bead beater, which breaks up bacterial cell wall mechanically by vibrating bacteria with microbeads at high speed. The effectiveness of this method for lysing M. paratuberculosis cells was compared to that of the freeze-thaw method, and use of commercial kits such as the InstaGene Matrix and the QIAamp Tissue Kit. The bead beater procedure was tested in combination with various cell lysis and template DNA preparation procedures to determine which of these steps improved the limit of detection of PCR assay that amplifies a 413 bp fragment of the IS900 gene. Results showed that the use of the bead beater, in combination with the use of lysis buffer, boiling, and isopropanol precipitation, decreased the limit of detection of M. paratuberculosis in milk by the PCR to 10(2) cfu/mL. The limit of detection was further decreased to 10 cfu/mL when 0.0037% bovine serum albumin was included in the PCR reaction mixtures. The improved assay was 10- to 10(4)-fold more sensitive than the PCR assays using template DNA prepared by other lysis procedures including boiling alone, freeze-thaw plus boiling, or use of commercial kits for lysis. PMID:11768125

  20. Bovine paratuberculosis III. An evaluation of a whole blood lymphocyte transformation test.

    OpenAIRE

    de Lisle, G W; Duncan, J R

    1981-01-01

    A whole blood lymphocyte transformation test was used to examine cattle with varying degrees of infection with Mycobacterium paratuberculosis. Minimally infected animals characteristically responded to johnin purified protein derivative in the lymphocyte transformation test but did not routinely react on serological and/or skin testing. Heavily infected animals showed considerable variation in their lymphocyte transformation responses to antigen and some of them were consistently unresponsive...

  1. Paratuberculosis on small ruminant dairy farms in Ontario, Canada: A survey of management practices.

    Science.gov (United States)

    Bauman, Cathy A; Jones-Bitton, Andria; Menzies, Paula; Jansen, Jocelyn; Kelton, David

    2016-05-01

    A cross-sectional study was undertaken (October 2010 to August 2011) to determine the risk factors for dairy goat herds and dairy sheep flocks testing positive for paratuberculosis (PTB) in Ontario, Canada. A questionnaire was administered to 50 producers during a farm visit in which concurrently, 20 randomly selected, lactating animals over the age of 2 years underwent sampling for paratuberculosis testing. Only 1 of 50 farms (2.0%) was closed to animal movement, whereas 96.6% of dairy goat farms and 94.1% of sheep farms purchased livestock from other producers. Only 10.3% of dairy goat, and no dairy sheep farms used artificial insemination. Manure was spread on grazing pastures by 65.5% and 70.6% of dairy goat and dairy sheep farms, respectively. Because of the high true-prevalence of paratuberculosis infection detected, no risk factor analysis could be performed. This study demonstrates that biosecurity practices conducive to transmission of PTB are highly prevalent in Ontario small ruminant dairy farms. PMID:27152042

  2. Utilization of a ts-sacB selection system for the generation of a Mycobacterium avium serovar-8 specific glycopeptidolipid allelic exchange mutant

    OpenAIRE

    Belisle John T; Inamine Julia M; Eckstein Torsten M; Lee Sun-Hwa; Irani Vida R; Maslow Joel N

    2004-01-01

    Abstract Background Mycobacterium avium are ubiquitous environmental organisms and a cause of disseminated infection in patients with end-stage AIDS. The glycopeptidolipids (GPL) of M. avium are proposed to participate in the pathogenesis of this organism, however, establishment of a clear role for GPL in disease production has been limited by the inability to genetically manipulate M. avium. Methods To be able to study the role of the GPL in M. avium pathogenesis, a ts-sacB selection system,...

  3. [Isolation of Mycobacterium avium-intracellulare from a hepatic biopsy].

    Science.gov (United States)

    Ruiz, Aroldo; Mederos, Lilian; Capó, Virginia

    2002-01-01

    A 64 years-old patient, who was a farmer suffering from chronic fever for two years, loss of weight and acute asthenia, was studied. He was admitted to "Pedro Kourí" Tropical Medicine Institute where the studies were conducted and revealed a globular sedimentation rate of 116 mm in 2 hours, and anemia of 9,8g% hemoglobin. The laparoscopic study indicated hepatic granulomatosis that was confirmed by hepatic biopsy in which a sample was taken from the liver to be microbiologically and cytologically examined. By microbiological methods, a non-pigmented slowly-growing strain was isolated, which was classified by conventional diagnostic techniques for the non-tuberculous mycobacteria classification and the alternative diagnosing technique known as bidimensional thin layer chromatography to confirm the previous classification and set the mycolic acid patterns. The isolated strain belonged to group III of Rynyon and was identified as Mycobacterium avium-intracellulare. PMID:15849945

  4. Mycobacterium avium complex enteritis in HIV-infected patient

    Directory of Open Access Journals (Sweden)

    Masahiro Ishikane

    2014-01-01

    Full Text Available Disseminated Mycobacterium avium complex (MAC infection is an important AIDS-defining opportunistic infection. The introduction of antimicrobial prophylaxis and antiretroviral therapy (ART markedly reduced the incidence of disseminated MAC infection and improved the survival of affected individuals. However, it seems that patients with new or recurrent MAC infection are still encountered in clinical practice. Our images captured the characteristic endoscopic findings of MAC duodenitis. The gastrointestinal (GI tract appears to be a common port of entry for MAC infection in patients with AIDS. Early recognition of GI MAC infection by endoscopy in HIV-infected patients and initiation of anti-MAC therapy and ART may reduce morbidity and mortality.

  5. 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. PMID:27111397

  6. 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. PMID:8858587

  7. 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. PMID:20467849

  8. Searching for proteins of subspecies with diagnostic potential by comparative qualitative proteomic analysis of mycobacterial tuberculins

    OpenAIRE

    Santema, Wiebren; Overdijk, Marije; Barends, Judith; Krijgsveld, Jeroen; Rutten, Victor; Koets, Ad

    2009-01-01

    Abstract Accurate immunodiagnosis of bovine paratuberculosis is among others hampered by the lack of specific antigens. One of the most frequently used antigen preparations is purified protein derivative (PPD), also known as tuberculin. This crude extract has limitations when used in diagnostic assays due to the presence of cross-reactive antigens. The aim of the current study was to systematically analyze the qualitative protein composition of PPD of the major mycobacterial pathog...

  9. Isolation and Characterization of Coproporphyrin Produced by Four Subspecies of Bacillus thuringiensis

    OpenAIRE

    Harms, R. L.; Martinez, D. R.; Griego, V M

    1986-01-01

    It was found by using spectrophotometric, spectrofluorometric, and high-pressure liquid chromatography that four subspecies of Bacillus thuringiensis produce coproporphyrin. The porphyrin isomer was identified as coproporphyrin I for B. thuringiensis subsp. kurstaki (HD1). The porphyrin was isolated both from spores and from a variety of spent growth media. The quantity of porphyrin released by each Bacillus subspecies differed. The rank order of porphyrin production follows: B. thuringiensis...

  10. Molecular and phenotypic distinction of the very recently evolved insular subspecies Mus musculus helgolandicus ZIMMERMANN, 1953

    OpenAIRE

    Babiker, Hiba; Tautz, Diethard

    2015-01-01

    Background Populations and subspecies of the house mouse Mus musculus were able to invade new regions worldwide in the wake of human expansion. Here we investigate the origin and colonization history of the house mouse inhabiting the small island of Heligoland on the German Bight - Mus musculus helgolandicus. It was first described by Zimmermann in 1953, based on morphological descriptions which were considered to be a mosaic between the subspecies M. m. domesticus and M. m. musculus. Since m...

  11. Two new subspecies of Lasianthus inodorus (Rubiaceae) from Kinabalu, Borneo, and their biogeographical implication

    OpenAIRE

    Zhu, Hua

    2001-01-01

    Two populations from Mount Kinabalu, Borneo, are recognised as two new subspecies of Lasianthus inodorus Blume (Rubiaceae), which occurs in montane habitats in mainland Southeast Asia, Sumatra and Java. The species and its subspecies are considered to compose a particular taxonomic group in the genus. Ecology and biogeography of the species group are discussed with the historical explanation of the tectonic history of Cenozoic Southeast Asia. The example strongly supports the concept of flori...

  12. Adhesion of Campylobacter jejuni and Mycobacterium avium onto polyethylene terephtalate (PET) used for bottled waters.

    Science.gov (United States)

    Tatchou-Nyamsi-König, Josiane-Aurore; Dague, Etienne; Mullet, Martine; Duval, Jérôme F L; Gaboriaud, Fabien; Block, Jean-Claude

    2008-12-01

    Adhesion of the bacteria Campylobacter jejuni and Mycobacterium avium onto polyethylene terephtalate (PET), a polymer widely used within the bottled water industry was measured in two different groundwater solutions. From this, it was found that whilst the percentage cell adhesion for a given strain did not change between groundwater types, substantial variation was obtained between the two bacterial species tested: M. avium (10-30% adhered cells) and C. jejuni (1-2%) and no major variations were measured as a function of groundwater composition for a given strain. To explain this, the interfacial electro-hydrodynamic properties of the bacteria were investigated by microelectrophoresis, with the resultant data analysed on the basis of electrokinetic theory for soft biocolloidal particles. The results obtained showed that M. avium carries a significant volume charge density and that its peripheral layer exhibits limited hydrodynamic flow permeation compared to that of C. jejuni. It was also demonstrated that steric hindrance to flow penetration and the degree of hydrophobicity within/of the outer bacterial interface are larger for M. avium cells. In line with this, the larger amount of M. avium cells deposited onto PET substrates as compared to that of C. jejuni can be explained by hydrophobic attraction and chemical binding between hydrophobic PET and outer soft surface layer of the bacteria. Hydrophobicity of PET was addressed by combining contact angle analyses and force spectroscopy using CH(3)-terminated AFM tip. PMID:18929388

  13. A novel multi-stage subunit vaccine against paratuberculosis induces significant immunity and reduces bacterial burden in tissues (P4304)

    DEFF Research Database (Denmark)

    Thakur, Aneesh; Aagaard, Claus; Riber, Ulla;

    2013-01-01

    Effective control of paratuberculosis is hindered by lack of a vaccine preventing infection, transmission and without diagnostic interference with tuberculosis. We have developed a novel multi-stage recombinant subunit vaccine in which a fusion of four early expressed MAP antigens is combined with...... characterized by a significant containment of bacterial burden in gut tissues compared to non-vaccinated animals. There was no cross-reaction with bovine tuberculosis in vaccinated animals. This novel multi-stage vaccine has the potential to become a marker vaccine for paratuberculosis....

  14. Burial increases seed longevity of two Artemisia tridentata (Asteraceae) subspecies

    Science.gov (United States)

    Wijayratne, Upekala C.; Pyke, David A.

    2012-01-01

    Premise of the study: Seed longevity and persistence in soil seed banks may be especially important for population persistence in ecosystems where opportunities for seedling establishment and disturbance are unpredictable. The fire regime, an important driver of population dynamics in sagebrush steppe ecosystems, has been altered by exotic annual grass invasion. Soil seed banks may play an active role in postfire recovery of the foundation shrub Artemisia tridentata, yet conditions under which seeds persist are largely unknown. Methods: We investigated seed longevity of two Artemisia tridentata subspecies in situ by retrieving seed bags that were placed at varying depths over a 2 yr period. We also sampled naturally dispersed seeds in litter and soil immediately after seed dispersal and before flowering in subsequent seasons to estimate seed persistence. Key results: After 24 mo, seeds buried at least 3 cm below the soil surface retained 30–40% viability whereas viability of seeds on the surface and under litter declined to 0 and Artemisia tridentata has the potential to form a short-term soil seed bank that persists longer than has been commonly assumed, and that burial is necessary for seed longevity. Use of seeding techniques that promote burial of some seeds to aid in formation of a soil seed bank may increase restoration potential.

  15. Chromosome painting defines genomic rearrangements between red howler monkey subspecies.

    Science.gov (United States)

    Consigliere, S; Stanyon, R; Koehler, U; Agoramoorthy, G; Wienberg, J

    1996-06-01

    We hybridized whole human chromosome-specific DNA libraries to chromosomes of two supposed subspecies of Alouatta seniculus: Alouatta seniculus sara and Alouatta seniculus arctoides. The number of hybridization signals per haploid set is 42 in A. s. sara and 43 in A. s. arctoidea; the two karyotypes differ by at least 16 chromosomal rearrangements, including numerous translocations. An unusual sex chromosome system is shared by both taxa. The sex chromosome system results from a Y translocation with a chromosome homologous to parts of human chromosome 3/15 and can be described as X1X2Y1Y2/X1X1X2X2 (male/female). Both red howlers also have microchromosomes, a highly unusual karyological trait not found in other higher primates. These microchromosomes are not hybridized by any human chromosome paint and therefore are probably composed of repetitive DNA. It is well known that New World monkeys have high karyological variability. It is probable that molecular cytogenetic analyses including chromosome painting will permit an accurate reconstruction of the phylogeny of these monkeys and help establish the ancestral karyotype for higher primates. PMID:8817065

  16. 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

  17. Mycobacterium avium-intracellulare brain abscess in HIV-positive patient

    Science.gov (United States)

    Karne, Sampada S.; Sangle, Shashikala A.; Kiyawat, Dilip S.; Dharmashale, Sujata N.; Kadam, Dilip B.; Bhardwaj, Renu S.

    2012-01-01

    Mycobacterial opportunistic infections are a major cause of morbidity and mortality among patients living with HIV (PLHIV) worldwide. Nontuberculous mycobacterial (NTM) infection is one of the leading causes of opportunistic infection in patients with advanced acquired immunodeficiency syndrome i.e., with CD4 count less than 50/cu.mm. Mycobacterium avium complex (MAC) is among the most common opportunistic bacterial infections in those patients with advanced immunodeficiency apart from cryptococcal meningitis, progressive multifocal leukoencephalopathy, etc. Common presentations of mycobacterium avium complex are fever, lymphadenitis and respiratory disease. Immune reconstitution disease is also known to manifest with MAC infections in PLHIV on highly active antiretroviral therapy. Very few cases of central nervous system involvement due to NTM infection have been described. We are reporting a case of advanced acquired immunodeficiency who presented with brain abscess due to Mycobacterium avium intracellulare. PMID:22412276

  18. Mycobacterium avium-intracellulare brain abscess in HIV-positive patient

    Directory of Open Access Journals (Sweden)

    Sampada S Karne

    2012-01-01

    Full Text Available Mycobacterial opportunistic infections are a major cause of morbidity and mortality among patients living with HIV (PLHIV worldwide. Nontuberculous mycobacterial (NTM infection is one of the leading causes of opportunistic infection in patients with advanced acquired immunodeficiency syndrome i.e., with CD4 count less than 50/cu.mm. Mycobacterium avium complex (MAC is among the most common opportunistic bacterial infections in those patients with advanced immunodeficiency apart from cryptococcal meningitis, progressive multifocal leukoencephalopathy, etc. Common presentations of mycobacterium avium complex are fever, lymphadenitis and respiratory disease. Immune reconstitution disease is also known to manifest with MAC infections in PLHIV on highly active antiretroviral therapy. Very few cases of central nervous system involvement due to NTM infection have been described. We are reporting a case of advanced acquired immunodeficiency who presented with brain abscess due to Mycobacterium avium intracellulare.

  19. Salmonella virulence plasmid. Modular acquisition of the spv virulence region by an F-plasmid in Salmonella enterica subspecies I and insertion into the chromosome of subspecies II, IIIa, IV and VII isolates.

    OpenAIRE

    Boyd, E.F.; Hartl, D. L.

    1998-01-01

    The spv operon is common to all Salmonella virulence plasmids. DNA hybridization analysis indicates that the spv region is limited in distribution to serovars of Salmonella enterica subspecies I, II, IIIa, IV, and VII and is absent from Salmonella bongori isolates. Among strains of subspecies II, IIIa, and VII, all isolates examined contained sequences that hybridized with the spv region. However, among isolates of subspecies I, DNA sequences capable of hybridizing with the spv region were fo...

  20. The effect of Cerasus avium stalk extract on albumin glycation reaction

    Directory of Open Access Journals (Sweden)

    Mohadeseh Abdoli

    2014-10-01

    Full Text Available Background: Non-enzymatic glycosylation of proteins is the major cause of diabetic complications. The inhibition of glycation process can reduce complications of diabetes. In the Iranian traditional medicine, the decoction (boiled extraction of Cerasus avium stalk is used as a hypoglycemic agent. The aim of this study was to investigate the in vitro inhibitory effects of decoction and ethanolic and aqueous extracts of Cerasus avium stalk on albumin glycation reaction. Methods: In this experimental study, first, the ethanolic, aqueous and decoction extracts of Cerasus avium stalk were prepared. Then, different concentrations of these extracts were prepared and added to albumin and glucose solutions. Finally, compared to control group that was not treated with any extracts, the albumin glycation rate in the groups treated with various concentrations of extracts was evaluated using TBA (thio-barbituric acid method. Results: The results showed that compared to control group, decoction of Cerasus avium stalk in the concentrations of 20, 10 and 2 mg/dl could reduce albumin glycation to 85.10±1.55, 72.35±1.75 and 51.25±1.22 %, respectively (P>0.001. Moreover, in the concentration of 20 mg/dl, the inhibitory effect of decoction of Cerasus avium stalk on the albumin glycation reaction was higher than those of aqueous (P=0.021 and ethanolic (P=0.009 extracts. Conclusion: The findings showed that the extracs of Cerasus avium stalk, in particular in the decoction form, could significantly reduce the rate of albumin glycation; therefore, it can be used for decreasing diabetes mellitus complications.

  1. Responses to playback of different subspecies songs in the Reed Bunting (Emberiza s. schoeniclus)

    DEFF Research Database (Denmark)

    Matessi, Giuliano; Dabelsteen, Torben; Pilastro, A.

    2000-01-01

    Populations of Reed Buntings Emberiza schoeniclus in the western Palearctic are classified in two major subspecies groups according to morphology: northern migratory schoeniclus and Mediterranean resident intermedia. Songs of the two groups differ mainly in complexity and syllable structure, with...... intermedia songs being more complex. We explored the possibilities of song as a subspecies isolating mechanism by testing if male schoeniclus Reed Buntings reacted differently to field playbacks of songs from their own subspecies group, from the foreign subspecies group and from a control species, the...... Yellowhammer Emberiza citrinella. We tested 30 different males, each with a different song belonging to one of the three categories: own, foreign and Yellowhammer. Unlike songs of the two subspecies groups, Yellowhammer song elicited no response. Differences between reactions to own and foreign subspecies...

  2. Development of a HACCP-based approach to control paratuberculosis in infected Irish dairy herds.

    Science.gov (United States)

    McAloon, Conor G; Whyte, Paul; More, Simon J; O'Grady, Luke; Doherty, Michael L

    2015-06-15

    Paratuberculosis is a challenging disease to control at farm level, in part due to the poor sensitivity of diagnostic tests and a prolonged incubation period. Simulation studies have highlighted on-farm management to be the most important factor in preventing on-farm spread. A risk assessment (RA) and management plan (MP) approach (collectively, RAMP) has been adopted around the world as the most appropriate method of controlling disease in infected farms. However, there are problems with RAMP that remain to be resolved. The RA relies heavily on farmer recollection and estimation resulting in subjectivity and substantial inter-observer variability. MPs consist of a series of qualitative, farm specific recommendations showing how management can be improved. However, MP assessment is generally conducted informally, and progress is monitored through 'end-point' diagnostic testing of adult animals and repeated risk assessments. Hazard analysis and critical control point (HACCP) has been developed as a proactive alternative to end-point testing. We hypothesise that farm-based HACCP systems may be a useful alternative to RAMP on farms where more intensive monitoring and evaluation of controls for paratuberculosis is required. Therefore, the objective of this methodological study was to develop a HACCP-based system for paratuberculosis control. Critical control points (CCPs) relating to peri-parturient area management, calving, new-born calf management and colostrum management were identified as areas where additional control could be exerted above existing methods. Novel monitoring systems were developed for each CCP, along with targets and corrective actions. This system is intended for use in high prevalence herds, or farms where more robust monitoring of key control points may be beneficial. It is currently being trialled on infected commercial dairy herds in Ireland. PMID:25944176

  3. Latent infections are the most frequent form of paratuberculosis in slaughtered Friesian cattle

    Directory of Open Access Journals (Sweden)

    Patricia Vazquez

    2014-10-01

    Full Text Available Paratuberculosis is a chronic mycobacterial infection causing granulomatous enteritis in ruminants, whose pathogenesis and epidemiology poses numerous challenges, including latency and reactivation. The most recent and complete classification of paratuberculosis immunopathological types in cattle recognized five categories. In this study, 1031 slaughtered Friesian cattle were submitted to serological, microbiological and pathological examinations with the aim of maximizing the rate of case detection. In most cases, infected animals had minimal lesions and almost no other proof of infection (38.9%, while the more characteristic types with the whole constellation of microbiological and immunological evidences accounted for a lower proportion (7.7%. As these findings in cattle suggest similarities with the epidemiology of tuberculosis in humans, we propose to re-group the original immunopathological types into two broader paratuberculosis epidemio-pathogenic forms or states: latent and patent. The former term would define infections with focal lesions and might constitute an apparent resilience status representing a difficult to detect reservoir of infection whose role could become critical if later immune-compromising factors lead to re-activation. The latter would group those cases with multifocal and diffuse inflammatory lesions with higher mycobacterial load and viability suggestive of a more immediate epidemiological risk. Interestingly, the relative frequency of presentation of each profile varied with age. The proportion of latent forms remained relatively constant between 33.8% and 54.3% through adulthood from 3 years of age, while patent forms were more frequent during the first years of age and tended to decrease among the oldest individuals.

  4. Mycobacterium avium in pygmy rabbits (Brachylagus idahoensis): 28 cases.

    Science.gov (United States)

    Harrenstien, Lisa A; Finnegan, Mitchell V; Woodford, Nina L; Mansfield, Kristin G; Waters, W Ray; Bannantine, John P; Paustian, Michael L; Garner, Michael M; Bakke, Antony C; Peloquin, Charles A; Phillips, Terry M

    2006-12-01

    The Columbia basin subpopulation of pygmy rabbit Brachylagus idahoensis was listed as endangered by the United States Fish and Wildlife Service in November 2001, and no pygmy rabbits have been seen in the wild since spring 2002. Captive propagation efforts have attempted to increase population size in preparation for reintroduction of animals into central Washington. Disseminated mycobacteriosis due to Mycobacterium avium has been the most common cause of death of adult captive pygmy rabbits. Between June 2002 and September 2004, mycobacteriosis was diagnosed in 28 captive adult pygmy rabbits (representing 29% of the captive population), in contrast to 18 adult pygmy rabbits dying of all other causes in the same time period. Antemortem and postmortem medical records were evaluated retrospectively to describe the clinical course of mycobacteriosis in pygmy rabbits, physical examination findings, and diagnostic test results in the diagnosis of mycobacteriosis in pygmy rabbits. Various treatment protocols, possible risk factors for mortality, and recommendations for prevention of mycobacteriosis were evaluated also. Compromised cell-mediated immunity appears to be the best explanation at this time for the observed high morbidity and mortality from mycobacterial infections in pygmy rabbits. PMID:17315435

  5. Analytical Verification of a PCR Assay for Identification of Bordetella avium

    OpenAIRE

    Register, Karen B.; Yersin, Andrew G.

    2005-01-01

    Bordetella avium is the etiologic agent of turkey coryza or bordetellosis, a respiratory disease responsible for substantial economic losses to the turkey industry. At present, identification of this bacterium relies on isolation and biochemical testing. Although a PCR for the detection of B. avium was proposed a number of years ago (P. H. Savelkoul, L. E. de Groot, C. Boersma, I. Livey, C. J. Duggleby, B. A. van der Zeijst, and W. Gaastra, Microb. Pathog. 15:207-215, 1993), lack of analytica...

  6. Organic Turkey Flocks: A Reservoir of Streptococcus gallolyticus subspecies gallolyticus.

    Directory of Open Access Journals (Sweden)

    Jochen Schulz

    Full Text Available Streptococcus gallolyticus subspecies gallolyticus (S. gallolyticus can colonise the gastrointestinal tract of humans and animals and is known to cause similar infections in both humans and animals. Data about the spread or prevalence in farm animals are missing. In this study, Trypton Soya Agar was modified to a selective medium enabling the isolation and quantification of S. gallolyticus from faecal samples. The bacterium was observed in 82 out of 91 faecal samples obtained from 18 different organic turkey flocks. The prevalence of shedding birds was estimated by the number of positive fresh droppings and reached up to 100% on most farms. Furthermore, for the first time S. gallolyticus was quantified in faeces from poultry flocks. The median of colony forming units (CFU per gramme faeces was 3.6 x 10(5 CFU/g. Typing of one isolate from each positive faecal sample by multilocus sequence typing delivered 24 sequence types (STs. Most of the isolates belonged to the clonal complex CC58. The same STs of this complex were detected in up to six different flocks. Partly, these flocks were located in various regions and stocked with varying breeding lines. Regarding the biochemical profiles of the same STs from different farms, the results did not contradict a spread of specific STs in the organic turkey production. Moreover, checking the pubMLST database revealed that STs found in this study were also found in other animal species and in humans. The high detection rate and the number of S. gallolyticus in turkey faeces indicate that this bacterium probably belongs to the common microbiota of the gastrointestinal tract of turkeys from organic flocks. Furthermore, the findings of this study support the suggestion of a possible interspecies transmission.

  7. A distinctive new subspecies of Scytalopus griseicollis ( Aves , Passeriformes , Rhinocryptidae ) from the northern Eastern Cordillera of Colombia and Venezuela

    OpenAIRE

    Jorge Avendaño; Donegan, Thomas M.

    2015-01-01

    Abstract We describe a new subspecies of Pale-bellied Tapaculo Scytalopus griseicollis from the northern Eastern Cordillera of Colombia and Venezuela. This form differs diagnosably in plumage from described subspecies Scytalopus griseicollis griseicollis and Scytalopus griseicollis gilesi and from the latter in tail length. It is also differentiated non-diagnosably in voice from both these populations. Ecological niche modelling analysis suggests that the new subspecies is restricted to the A...

  8. Interpretation of the Gamma Interferon Test for Diagnosis of Subclinical Paratuberculosis in Cattle

    OpenAIRE

    Jungersen, G; Huda, A; Hansen, J. J.; Lind, P.

    2002-01-01

    A group of 252 cattle without clinical signs of paratuberculosis (paraTB) in 10 herds infected with paraTB and a group of 117 cattle in 5 herds without paraTB were selected. Whole-blood samples were stimulated with bovine, avian, and johnin purified protein derivative (PPD) and examined for gamma interferon (IFN-γ) release. For diagnosis of paraTB, satisfactory estimated specificities (95 to 99%) could be obtained by johnin PPD stimulation irrespective of interpretation relative to bovine PPD...

  9. Shift of grey seal subspecies boundaries in response to climate, culling and conservation.

    Science.gov (United States)

    Fietz, Katharina; Galatius, Anders; Teilmann, Jonas; Dietz, Rune; Frie, Anne Kristine; Klimova, Anastasia; Palsbøll, Per J; Jensen, Lasse F; Graves, Jeff A; Hoffman, Joseph I; Olsen, Morten Tange

    2016-09-01

    Identifying the processes that drive changes in the abundance and distribution of natural populations is a central theme in ecology and evolution. Many species of marine mammals have experienced dramatic changes in abundance and distribution due to climatic fluctuations and anthropogenic impacts. However, thanks to conservation efforts, some of these species have shown remarkable population recovery and are now recolonizing their former ranges. Here, we use zooarchaeological, demographic and genetic data to examine processes of colonization, local extinction and recolonization of the two northern European grey seal subspecies inhabiting the Baltic Sea and North Sea. The zooarchaeological and genetic data suggest that the two subspecies diverged shortly after the formation of the Baltic Sea approximately 4200 years bp, probably through a gradual shift to different breeding habitats and phenologies. By comparing genetic data from 19th century pre-extinction material with that from seals currently recolonizing their past range, we observed a marked spatiotemporal shift in subspecies boundaries, with increasing encroachment of North Sea seals on areas previously occupied by the Baltic Sea subspecies. Further, both demographic and genetic data indicate that the two subspecies have begun to overlap geographically and are hybridizing in a narrow contact zone. Our findings provide new insights into the processes of colonization, extinction and recolonization and have important implications for the management of grey seals across northern Europe. PMID:27616353

  10. Beta2-adrenergic receptor stimulation inhibits nitric oxide generation by Mycobacterium avium infected macrophages.

    Science.gov (United States)

    Boomershine, C S; Lafuse, W P; Zwilling, B S

    1999-11-01

    Catecholamine regulation of nitric oxide (NO) production by IFNgamma-primed macrophages infected with Mycobacterium avium was investigated. Epinephrine treatment of IFNgamma-primed macrophages at the time of M. avium infection inhibited the anti-mycobacterial activity of the cells. The anti-mycobacterial activity of macrophages correlated with NO production. Using specific adrenergic receptor agonists, the abrogation of mycobacterial killing and decreased NO production by catecholamines was shown to be mediated via the beta2-adrenergic receptor. Elevation of intracellular cAMP levels mimicked the catecholamine-mediated inhibition of NO in both M. avium infected and LPS stimulated macrophages. Specific inhibitors of both adenylate cyclase and protein kinase A prevented the beta2-adrenoceptor-mediated inhibition of nitric oxide production. Beta2-adrenoreceptor stimulation at the time of M. avium infection of IFNgamma-primed macrophages also inhibited expression of iNOS mRNA. These observations show that catecholamine hormones can affect the outcome of macrophage-pathogen interactions and suggest that one result of sympathetic nervous system activation is the suppression of the capacity of macrophages to produce anti-microbial effector molecules. PMID:10580815

  11. Activity of Picolinic Acid in Combination with the Antiprotozoal Drug Quinacrine against Mycobacterium avium Complex

    OpenAIRE

    Shimizu, Toshiaki; Tomioka, Haruaki

    2006-01-01

    We studied the in vitro and in vivo antimicrobial activities of picolinic acid (PA) in combination with the antiprotozoal drug quinacrine against intramacrophage Mycobacterium avium complex (MAC). Quinacrine significantly potentiated the anti-MAC activity of PA, suggesting the usefulness of this combination in the clinical control of MAC infection.

  12. 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")

  13. Alternaria cerasidanica sp nov., isolated in Denmark from drupes of Prunus avium

    DEFF Research Database (Denmark)

    Roberts, R. G.; Reymond, S. T.; Andersen, Birgitte

    2010-01-01

    The ex-type strain of Alternaria cerasidanica was isolated in 2001 from an immature, asymptomatic drupe of Prunus avium collected at a commercial cherry orchard near Skaelskor, Denmark. Cultural morphology, sporulation pattern and cluster analyses of combined RAPD, RAMS (microsatellite), and AFLP...

  14. Analysis of wild sweet cherry (Prunus avium L.) germplasm diversity in south-east Serbia

    OpenAIRE

    Mratinić Evica; Fotirić-Akšić Milica; Jovković Radmila

    2012-01-01

    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 ade...

  15. Protection of mice from Mycobacterium avium infection by recombinant interleukin-12.

    OpenAIRE

    Kobayashi, K; Kasama, T.; Yamazaki, J.; Hosaka, M; Katsura, T.; Mochizuki, T; Soejima, K; Nakamura, R M

    1995-01-01

    Treatment with interleukin-12 (IL-12) significantly reduced the number of viable bacteria in mice infected with Mycobacterium avium. IL-12 itself, however, could not inhibit directly mycobacterial growth in vitro. IL-12 exerts antimycobacterial activity in vivo with a low level of toxicity, possibly by enhancing the host defense against the infection.

  16. Molecular Analysis of Mycobacterium avium Isolates by Using Pulsed-Field Gel Electrophoresis and PCR

    Science.gov (United States)

    Pestel-Caron, Martine; Graff, Gabriel; Berthelot, Gilles; Pons, Jean-Louis; Lemeland, Jean-François

    1999-01-01

    Genetic relationships among 46 isolates of Mycobacterium avium recovered from 37 patients in a 2,500-bed hospital from 1993 to 1998 were assessed by pulsed-field gel electrophoresis (PFGE) and PCR amplification of genomic sequences located between the repetitive elements IS1245 and IS1311. Each technique enabled the identification of 27 to 32 different patterns among the 46 isolates, confirming that the genetic heterogeneity of M. avium strains is high in a given community. Furthermore, this retrospective analysis of sporadic isolates allowed us (i) to suggest the existence of two remanent strains in our region, (ii) to raise the question of the possibility of nosocomial acquisition of M. avium strains, and (iii) to document laboratory contamination. The methods applied in the present study were found to be useful for the typing of M. avium isolates. In general, both methods yielded similar results for both related and unrelated isolates. However, the isolates in five of the six PCR clusters were distributed among two to three PFGE patterns, suggesting that this PCR-based method may have limitations for the analysis of strains with low insertion sequence copy numbers or for resolution of extended epidemiologic relationships. PMID:10405383

  17. 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.

  18. Genetic subspecies diversity of the chimpanzee CD4 virus-receptor gene

    DEFF Research Database (Denmark)

    Hvilsom, Christina; Carlsen, Frands; Siegismund, Hans R;

    2008-01-01

    Chimpanzees are naturally and asymptomatically infected by simian immunodeficiency virus (SIV). Pathogenic properties of SIV/HIV vary and differences in susceptibility and pathogenicity of SIV/HIV depend in part on host-specific factors such as virus-receptor/co-receptor interactions. Since CD4...... plays a primary role in virus binding and since SIVcpz have been found only in two African chimpanzee subspecies, we characterized the genetic diversity of CD4 receptors in all four recognized subspecies of chimpanzees. We found noticeable variation in the first variable region V1 of CD4 and in intron...

  19. Characterization of a novel plasmid, pMAH135, from Mycobacterium avium subsp. hominissuis.

    Directory of Open Access Journals (Sweden)

    Kei-ichi Uchiya

    Full Text Available Mycobacterium avium complex (MAC causes mainly two types of disease. The first is disseminated disease in immunocompromised hosts, such as individuals infected by human immunodeficiency virus (HIV. The second is pulmonary disease in individuals without systemic immunosuppression, and the incidence of this type is increasing worldwide. M. avium subsp. hominissuis, a component of MAC, causes infection in pigs as well as in humans. Many aspects of the different modes of M. avium infection and its host specificity remain unclear. Here, we report the characteristics and complete sequence of a novel plasmid, designated pMAH135, derived from M. avium strain TH135 in an HIV-negative patient with pulmonary MAC disease. The pMAH135 plasmid consists of 194,711 nucleotides with an average G + C content of 66.5% and encodes 164 coding sequences (CDSs. This plasmid was unique in terms of its homology to other mycobacterial plasmids. Interestingly, it contains CDSs with sequence homology to mycobactin biosynthesis proteins and type VII secretion system-related proteins, which are involved in the pathogenicity of mycobacteria. It also contains putative conserved domains of the multidrug efflux transporter. Screening of isolates from humans and pigs for genes located on pMAH135 revealed that the detection rate of these genes was higher in clinical isolates from pulmonary MAC disease patients than in those from HIV-positive patients, whereas the genes were almost entirely absent in isolates from pigs. Moreover, variable number tandem repeats typing analysis showed that isolates carrying pMAH135 genes are grouped in a specific cluster. Collectively, the pMAH135 plasmid contains genes associated with M. avium's pathogenicity and resistance to antimicrobial agents. The results of this study suggest that pMAH135 influence not only the pathological manifestations of MAC disease, but also the host specificity of MAC infection.

  20. Structure and amount of genetic variation at minisatellite loci within the subspecies complex of Phoca vitulina (the harbour seal).

    NARCIS (Netherlands)

    A.L. Kappe; R. Bijlsma; A.D.M.E. Osterhaus (Albert); W. van Delden; L. van de Zande

    1997-01-01

    textabstractThe structure and amount of genetic variation within and between three subspecies of the harbour seal Phoca vitulina was assessed by multilocus DNA fingerprinting. Bandsharing similarity indicates that the subspecies Phoca vitulina richardsi (Alaska, East Pacific) is clearly separated fr

  1. 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 potentia...... as potting soil) and veterinary samples were found to contain viable M avium isolates belonging to genotypes also found in humans....

  2. 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.

  3. Accounting for uncertainty in model-based prevalence estimation: paratuberculosis control in dairy herds

    Directory of Open Access Journals (Sweden)

    Davidson Ross S

    2012-09-01

    Full Text Available Abstract Background A common approach to the application of epidemiological models is to determine a single (point estimate parameterisation using the information available in the literature. However, in many cases there is considerable uncertainty about parameter values, reflecting both the incomplete nature of current knowledge and natural variation, for example between farms. Furthermore model outcomes may be highly sensitive to different parameter values. Paratuberculosis is an infection for which many of the key parameter values are poorly understood and highly variable, and for such infections there is a need to develop and apply statistical techniques which make maximal use of available data. Results A technique based on Latin hypercube sampling combined with a novel reweighting method was developed which enables parameter uncertainty and variability to be incorporated into a model-based framework for estimation of prevalence. The method was evaluated by applying it to a simulation of paratuberculosis in dairy herds which combines a continuous time stochastic algorithm with model features such as within herd variability in disease development and shedding, which have not been previously explored in paratuberculosis models. Generated sample parameter combinations were assigned a weight, determined by quantifying the model’s resultant ability to reproduce prevalence data. Once these weights are generated the model can be used to evaluate other scenarios such as control options. To illustrate the utility of this approach these reweighted model outputs were used to compare standard test and cull control strategies both individually and in combination with simple husbandry practices that aim to reduce infection rates. Conclusions The technique developed has been shown to be applicable to a complex model incorporating realistic control options. For models where parameters are not well known or subject to significant variability, the

  4. Streptococcus equi subspecies zooepidemicus Infections in Humans by Zoonotic Transmission from Horses

    Centers for Disease Control (CDC) Podcasts

    2013-06-12

    Dr. Mike Miller reads an abridged version of the Emerging Infectious Diseases’ article, Streptococcus equi subspecies zooepidemicus Infections in Humans by Zoonotic Transmission from Horses.  Created: 6/12/2013 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 7/3/2013.

  5. Comparison of the bioluminescence of Photorhabdus species and subspecies type strains

    Czech Academy of Sciences Publication Activity Database

    Hyršl, P.; Číž, Milan; Lojek, Antonín

    2004-01-01

    Roč. 49, č. 5 (2004), s. 539. ISSN 0015-5632 R&D Projects: GA AV ČR IBS5004009 Institutional research plan: CEZ:AV0Z5004920 Keywords : bioluminescence * Photorhabdus species type strains * Photorhabdus subspecies type strains Subject RIV: BO - Biophysics Impact factor: 1.034, year: 2004

  6. Decoding regulatory landscape of somatic embryogenesis reveals differential regulatory networks between japonica and indica rice subspecies.

    Science.gov (United States)

    Indoliya, Yuvraj; Tiwari, Poonam; Chauhan, Abhisekh Singh; Goel, Ridhi; Shri, Manju; Bag, Sumit Kumar; Chakrabarty, Debasis

    2016-01-01

    Somatic embryogenesis is a unique process in plants and has considerable interest for biotechnological application. Compare to japonica, indica rice has been less responsive to in vitro culture. We used Illumina Hiseq 2000 sequencing platform for comparative transcriptome analysis between two rice subspecies at six different developmental stages combined with a tag-based digital gene expression profiling. Global gene expression among different samples showed greater complexity in japonica rice compared to indica which may be due to polyphyletic origin of two rice subspecies. Expression pattern in initial stage indicate major differences in proembryogenic callus induction phase that may serve as key regulator to observe differences between both subspecies. Our data suggests that phytohormone signaling pathways consist of elaborate networks with frequent crosstalk, thereby allowing plants to regulate somatic embryogenesis pathway. However, this crosstalk varies between the two rice subspecies. Down regulation of positive regulators of meristem development (i.e. KNOX, OsARF5) and up regulation of its counterparts (OsRRs, MYB, GA20ox1/GA3ox2) in japonica may be responsible for its better regeneration and differentiation of somatic embryos. Comprehensive gene expression information in the present experiment may also facilitate to understand the monocot specific meristem regulation for dedifferentiation of somatic cell to embryogenic cells. PMID:26973288

  7. Campylobacter fetus subspecies contain conserved type IV secretion systems on multiple genomic islands and plasmids

    Science.gov (United States)

    The features contributing to the differences in pathogenicity of the C. fetus subspecies are unknown. Putative factors involved in pathogenesis are located in genomic islands that encode type IV secretion system (T4SS) and fic-domain (filamentation induced by cyclic AMP) proteins. In the genomes of ...

  8. 77 FR 20773 - Endangered and Threatened Species; Proposed Threatened Status for Subspecies of the Ringed Seal

    Science.gov (United States)

    2012-04-06

    ...) subspecies of ringed seals as threatened (75 FR 77476). On December 13, 2011, in consideration of substantial... deadline for the final listing determination to June 10, 2012 (FR 77466). At that time, we also announced... the initial comment period on the December 10, 2011 (75 FR 77476), proposed rule should not...

  9. 76 FR 77466 - Endangered and Threatened Species; Proposed Threatened Status for Subspecies of the Ringed Seal

    Science.gov (United States)

    2011-12-13

    ... and 12-month finding for spotted seals in October 2009 (74 FR 53683, October 20, 2009; see also, 75 FR... ochotensis), and Ladoga (Phoca hispida ladogensis) subspecies of ringed seals as threatened (75 FR 77476). We... largha) under the ESA (73 FR 16617). On May 28, 2008, we received a petition from the Center...

  10. A new subspecies if the subgenus Ecdyonurus Eaton, 1868 (Ephemeroptera: Heptageniidae) from the East of Ukraine

    Czech Academy of Sciences Publication Activity Database

    Martynov, A. V.; Godunko, Roman J.

    2013-01-01

    Roč. 3666, č. 4 (2013), s. 489-509. ISSN 1175-5326 R&D Projects: GA ČR GA206/08/1389; GA MŠk(CZ) 7AMB12SK144 Institutional support: RVO:60077344 Keywords : new subspecies * description * distinguishing characters Subject RIV: EG - Zoology Impact factor: 1.060, year: 2013

  11. Application of mitochondrial DNA sequence analysis in the forensic identification of Chinese sika deer subspecies.

    Science.gov (United States)

    Wu, Hua; Wan, Qiu-Hong; Fang, Sheng-Guo; Zhang, Shu-Yan

    2005-03-10

    As a direct and indirect consequence of human activities, only two subspecies, Cervus nippon sinchuanicus and Cervus nippon kopschi, currently subsist in the wild of China. However, a large population of Cervus nippon hortulorum and Cervus nippon nippon is raised in order to gain deer parts for Chinese traditional medicine. According to Chinese Wild Animal Conservation Law, hunting, capturing and trading of the wild sika deer are strictly banned, however, raising and trading of the domestic individual are permitted. Thus, it is very necessary to identify the subspecies of sika deer in China in forensic tests. In our study, we used mitochondrial DNA control region sequence analysis and phylogenetic analysis to identify the subspecies of sika deer. Mitochondrial DNA control region sequences analysis revealed that two haplotypes came from the unknown samples. One is the same as the haplotype that came from the samples of wild population of C. n. kopschi. Phylogenetic analysis indicated that the two haplotypes of unknown samples clustered with the haplotypes of C. n. kopschi, and had significant difference from the haplotypes of the other subspecies. These results together revealed that the unknown samples came from two individuals that belong to the wild population of C. n. kopschi living in the Qinglingfeng State Natural Reserve of Zhejiang province. Therefore, the results provide forensic evidence of illegal wild animal hunting. PMID:15639603

  12. 77 FR 73769 - Endangered and Threatened Wildlife and Plants; Listing Four Subspecies of Mazama Pocket Gopher...

    Science.gov (United States)

    2012-12-11

    ... development and the operation of gravel pits (70 FR 24870; May 11, 2005) on gopher habitat. In our 2007 CNOR... subspecies meet the definition of an endangered or threatened species under the Act: Habitat loss through...; Inadequate existing regulatory mechanisms that allow significant threats such as habitat loss; and...

  13. Giemsa C-banding Karyotypes of Two Subspecies of Hordeum brevisubulatum from China

    DEFF Research Database (Denmark)

    Linde-Laursen, Ib; Bothmer, R.

    1984-01-01

    C-banding patterns ofH. brevisubulatum subsp.brevisubulatum (2x) and subsp.turkestanicum (4x) had conspicuous telomeric C-bands in at least one chromosome arm with a minor difference in average band size between subspecies. Other conspicuous bands were few in number as in other taxa of the specie...

  14. Mitogenomic Phylogenetics of Fin Whales (Balaenoptera physalus spp.) : Genetic Evidence for Revision of Subspecies

    NARCIS (Netherlands)

    Archer, Frederick I.; Morin, Phillip A.; Hancock-Hanser, Brittany L.; Robertson, Kelly M.; Leslie, Matthew S.; Berube, Martine; Panigada, Simone; Taylor, Barbara L.

    2013-01-01

    There are three described subspecies of fin whales (Balaenoptera physalus): B. p. physalus Linnaeus, 1758 in the Northern Hemisphere, B. p. quoyi Fischer, 1829 in the Southern Hemisphere, and a recently described pygmy form, B. p. patachonica Burmeister, 1865. The discrete distribution in the North

  15. 78 FR 54214 - Endangered and Threatened Wildlife and Plants; Removing Five Subspecies of Mazama Pocket Gopher...

    Science.gov (United States)

    2013-09-03

    ...We, the U.S. Fish and Wildlife Service (Service), remove five subspecies of Mazama pocket gopher (Tacoma, Brush Prairie, Shelton, Olympic, and Cathlamet) from the list of candidates for listing as threatened or endangered species under the Endangered Species Act of 1973, as amended. After review of the best available scientific and commercial information, we find that the Tacoma pocket gopher......

  16. Investigating concordance among genetic data, subspecies circumscriptions and hostplant use in the nymphalid butterfly Polygonia faunus.

    Directory of Open Access Journals (Sweden)

    Ullasa Kodandaramaiah

    Full Text Available Subspecies are commonly used taxonomic units to formally describe intraspecific geographic variation in morphological traits. However, the concept of subspecies is not clearly defined, and there is little agreement about what they represent in terms of evolutionary units, and whether they can be used as reliably useful units in conservation, evolutionary theory and taxonomy. We here investigate whether the morphologically well-characterized subspecies in the North American butterfly Polygonia faunus are supported by genetic data from mitochondrial sequences and eight microsatellite loci. We also investigate the phylogeographic structure of P. faunus and test whether similarities in host-plant use among populations are related to genetic similarity. Neither the nuclear nor the mitochondrial data corroborated subspecies groupings. We found three well defined genetic clusters corresponding to California, Arizona and (New Mexico+Colorado. There was little structuring among the remaining populations, probably due to gene flow across populations. We found no support for the hypothesis that similarities in host use are related to genetic proximity. The results indicate that the species underwent a recent rapid expansion, probably from two glacial refugia in western North America. The mitochondrial haplotype network indicates at least two independent expansion phases into eastern North America. Our results clearly demonstrate that subspecies in P. faunus do not conform to the structuring of genetic variation. More studies on insects and other invertebrates are needed to better understand the scope of this phenomenon. The results of this study will be crucial in designing further experiments to understand the evolution of hostplant utilization in this species.

  17. Population structure of honey bees in the Carpathian Basin (Hungary) confirms introgression from surrounding subspecies.

    Science.gov (United States)

    Péntek-Zakar, Erika; Oleksa, Andrzej; Borowik, Tomasz; Kusza, Szilvia

    2015-12-01

    Carniolan honey bees (Apis mellifera carnica) are considered as an indigenous subspecies in Hungary adapted to most of the ecological and climatic conditions in this area. However, during the last decades Hungarian beekeepers have recognized morphological signs of the Italian honey bee (Apis mellifera ligustica). As the natural distribution of the honey bee subspecies can be affected by the importation of honey bee queens or by natural gene flow, we aimed at determining the genetic structure and characteristics of the local honey bee population using molecular markers. All together, 48 Hungarian and 84 foreign (Italian, Polish, Spanish, Liberian) pupae and/or workers were used for mitochondrial DNA analysis. Additionally, 53 sequences corresponding to 10 subspecies and the Buckfast hybrid were downloaded from GenBank. For the nuclear analysis, 236 Hungarian and 106 foreign honey bees were genotyped using nine microsatellites. Heterozygosity values, population-specific alleles, FST values, principal coordinate analysis, assignment tests, structure analysis, and dendrograms were calculated. Haplotype and nucleotide diversity values showed moderate values. We found that one haplotype (H9) was dominant in Hungary. The presence of the black honey bee (Apis mellifera mellifera) was negligible, but a few individuals resembling other subspecies were identified. We proved that the Hungarian honey bee population is nearly homogeneous but also demonstrated introgression from the foreign subspecies. Both mitochondrial DNA and microsatellite analyses corroborated the observations of the beekeepers. Molecular analyses suggested that Carniolan honey bee in Hungary is slightly affected by Italian and black honey bee introgression. Genetic differences were detected between Polish and Hungarian Carniolan honey bee populations, suggesting the existence of at least two different gene pools within A. m. carnica. PMID:27069597

  18. Fine analysis of genetic diversity of the tpr gene family among treponemal species, subspecies and strains.

    Directory of Open Access Journals (Sweden)

    Arturo Centurion-Lara

    Full Text Available BACKGROUND: The pathogenic non-cultivable treponemes include three subspecies of Treponema pallidum (pallidum, pertenue, endemicum, T. carateum, T. paraluiscuniculi, and the unclassified Fribourg-Blanc treponeme (Simian isolate. These treponemes are morphologically indistinguishable and antigenically and genetically highly similar, yet cross-immunity is variable or non-existent. Although all of these organisms cause chronic, multistage skin and systemic disease, they have historically been classified by mode of transmission, clinical presentations and host ranges. Whole genome studies underscore the high degree of sequence identity among species, subspecies and strains, pinpointing a limited number of genomic regions for variation. Many of these "hot spots" include members of the tpr gene family, composed of 12 paralogs encoding candidate virulence factors. We hypothesize that the distinct clinical presentations, host specificity, and variable cross-immunity might reside on virulence factors such as the tpr genes. METHODOLOGY/PRINCIPAL FINDINGS: Sequence analysis of 11 tpr loci (excluding tprK from 12 strains demonstrated an impressive heterogeneity, including SNPs, indels, chimeric genes, truncated gene products and large deletions. Comparative analyses of sequences and 3D models of predicted proteins in Subfamily I highlight the striking co-localization of discrete variable regions with predicted surface-exposed loops. A hallmark of Subfamily II is the presence of chimeric genes in the tprG and J loci. Diversity in Subfamily III is limited to tprA and tprL. CONCLUSIONS/SIGNIFICANCE: An impressive sequence variability was found in tpr sequences among the Treponema isolates examined in this study, with most of the variation being consistent within subspecies or species, or between syphilis vs. non-syphilis strains. Variability was seen in the pallidum subspecies, which can be divided into 5 genogroups. These findings support a genetic basis for

  19. Development of a Multi-Stage Vaccine against Paratuberculosis in Cattle

    DEFF Research Database (Denmark)

    Thakur, Aneesh

    considerable economic losses to farming community. Paratuberculosis is a staged infection in which young calves acquire the infection in the first months of life, may progress into a prolonged asymptomatic stage of about 2-5 years and may eventually become clinically infected animals. Vaccination with whole......-cell live or inactivated vaccines prevents or delays the development of clinical stage of the disease but does not eliminate MAP and is usually accompanied by interference with bovine tuberculosis diagnostics as well as local tissue damage. Subunit vaccines with well-defined antigens in combination with a...... suitable adjuvant offer the possibility to avoid these limitations and induce a strong T helper 1 (TH1) type immune response that has been associated with protection against MAP. The aim of the study was to identify proteins from different stages of infection and formulate them into a multi-stage subunit...

  20. Interference of intradermal tuberculin tests on the serodiagnosis of paratuberculosis in cattle.

    Science.gov (United States)

    Varges, Renato; Marassi, Carla Dray; Oelemann, Walter; Lilenbaum, Walter

    2009-06-01

    In this experiment 63 animals from a paratuberculosis (PTB) and tuberculosis-free herd were tested by Intradermal Tuberculin Tests (ITT) and blood samples were collected before PPD inoculation and on days 3, 15, 30, 60 and 90 post-inoculation (p.i.). Sera were tested for PTB-specific antibodies by ELISA-PPA and confirmed by a commercial ELISA. Three (4.76%) animals were positive by ELISA-PPA and five (7.93%) in the commercial ELISA, between days 30 and 90 p.i. These results suggest that ITT can interfere in the reliability of ELISAs and that serological testing for PTB should be avoided for 90d after PPD inoculation. PMID:18930508

  1. A novel multi-stage subunit vaccine against paratuberculosis induces significant immunity and reduces bacterial burden in tissues (P4304)

    DEFF Research Database (Denmark)

    Thakur, Aneesh; Aagaard, Claus; Riber, Ulla; Skovgaard, Kerstin; Andersen, Peter; Jungersen, Gregers

    Effective control of paratuberculosis is hindered by lack of a vaccine preventing infection, transmission and without diagnostic interference with tuberculosis. We have developed a novel multi-stage recombinant subunit vaccine in which a fusion of four early expressed MAP antigens is combined with...... followed for a year. The FET11 vaccine induced a significant T cell response against constituent vaccine proteins characterized by a high percentage of CD4+ T cells and participation of polyfunctional CD4+ T cells. Of the two different age groups, late FET11 vaccination conferred protective immunity...... characterized by a significant containment of bacterial burden in gut tissues compared to non-vaccinated animals. There was no cross-reaction with bovine tuberculosis in vaccinated animals. This novel multi-stage vaccine has the potential to become a marker vaccine for paratuberculosis....

  2. Inactivation of Mycobacterium paratuberculosis and Mycobacterium tuberculosis in fresh soft cheese by gamma radiation

    Energy Technology Data Exchange (ETDEWEB)

    Badr, Hesham M., E-mail: heshambadr_aea@yahoo.co.uk [Atomic Energy Authority, Nuclear Research Center, Abou Zaabal, P.O. Box 13759 Cairo (Egypt)

    2011-11-15

    The effectiveness of gamma irradiation on the inactivation of Mycobacterium paratuberculosis, Mycobacterium bovis and Mycobacterium tuberculosis in fresh soft cheese that prepared from artificially inoculated milk samples was studied. Irradiation at dose of 2 kGy was sufficient for the complete inactivation of these mycobacteria as they were not detected in the treated samples during storage at 4{+-}1 {sup o}C for 15 days. Moreover, irradiation of cheese samples, that were prepared from un-inoculated milk, at this effective dose had no significant effects on their gross composition and contents from riboflavin, niacin and pantothenic acid, while significant decreases in vitamin A and thiamin were observed. In addition, irradiation of cheese samples had no significant effects on their pH and nitrogen fractions contents, except for the contents of ammonia, which showed a slight, but significant, increases due to irradiation. The analysis of cheese fats indicated that irradiation treatment induced significant increase in their oxidation parameters and contents from free fatty acids; however, the observed increases were relatively low. On the other hand, irradiation of cheese samples induced no significant alterations on their sensory properties. Thus, irradiation dose of 2 kGy can be effectively applied to ensure the safety of soft cheese with regards to these harmful mycobacteria. - Highlights: > We examined the effectiveness of gamma irradiation on inactivation of Mycobacterium paratuberculosis, Mycobacterium bovis and Mycobacterium tuberculosis in fresh soft cheese. > Irradiation at dose of 2 kGy was sufficient for complete inactivation of these mycobacteria. > Irradiation of cheese samples induced no significant alterations on their sensory properties.

  3. Inactivation of Mycobacterium paratuberculosis and Mycobacterium tuberculosis in fresh soft cheese by gamma radiation

    International Nuclear Information System (INIS)

    The effectiveness of gamma irradiation on the inactivation of Mycobacterium paratuberculosis, Mycobacterium bovis and Mycobacterium tuberculosis in fresh soft cheese that prepared from artificially inoculated milk samples was studied. Irradiation at dose of 2 kGy was sufficient for the complete inactivation of these mycobacteria as they were not detected in the treated samples during storage at 4±1 oC for 15 days. Moreover, irradiation of cheese samples, that were prepared from un-inoculated milk, at this effective dose had no significant effects on their gross composition and contents from riboflavin, niacin and pantothenic acid, while significant decreases in vitamin A and thiamin were observed. In addition, irradiation of cheese samples had no significant effects on their pH and nitrogen fractions contents, except for the contents of ammonia, which showed a slight, but significant, increases due to irradiation. The analysis of cheese fats indicated that irradiation treatment induced significant increase in their oxidation parameters and contents from free fatty acids; however, the observed increases were relatively low. On the other hand, irradiation of cheese samples induced no significant alterations on their sensory properties. Thus, irradiation dose of 2 kGy can be effectively applied to ensure the safety of soft cheese with regards to these harmful mycobacteria. - Highlights: → We examined the effectiveness of gamma irradiation on inactivation of Mycobacterium paratuberculosis, Mycobacterium bovis and Mycobacterium tuberculosis in fresh soft cheese. → Irradiation at dose of 2 kGy was sufficient for complete inactivation of these mycobacteria. → Irradiation of cheese samples induced no significant alterations on their sensory properties.

  4. Hybridization of Southern Hemisphere blue whale subspecies and a sympatric area off Antarctica: impacts of whaling or climate change?

    Science.gov (United States)

    Attard, Catherine R M; Beheregaray, Luciano B; Jenner, K Curt S; Gill, Peter C; Jenner, Micheline-Nicole; Morrice, Margaret G; Robertson, Kelly M; Möller, Luciana M

    2012-12-01

    Understanding the degree of genetic exchange between subspecies and populations is vital for the appropriate management of endangered species. Blue whales (Balaenoptera musculus) have two recognized Southern Hemisphere subspecies that show differences in geographic distribution, morphology, vocalizations and genetics. During the austral summer feeding season, the Antarctic blue whale (B. m. intermedia) is found in polar waters and the pygmy blue whale (B. m. brevicauda) in temperate waters. Here, we genetically analyzed samples collected during the feeding season to report on several cases of hybridization between the two recognized blue whale Southern Hemisphere subspecies in a previously unconfirmed sympatric area off Antarctica. This means the pygmy blue whales using waters off Antarctica may migrate and then breed during the austral winter with the Antarctic subspecies. Alternatively, the subspecies may interbreed off Antarctica outside the expected austral winter breeding season. The genetically estimated recent migration rates from the pygmy to Antarctic subspecies were greater than estimates of evolutionary migration rates and previous estimates based on morphology of whaling catches. This discrepancy may be due to differences in the methods or an increase in the proportion of pygmy blue whales off Antarctica within the last four decades. Potential causes for the latter are whaling, anthropogenic climate change or a combination of these and may have led to hybridization between the subspecies. Our findings challenge the current knowledge about the breeding behaviour of the world's largest animal and provide key information that can be incorporated into management and conservation practices for this endangered species. PMID:23137299

  5. Bayesian estimation of prevalence of paratuberculosis in dairy herds enrolled in a voluntary Johne's Disease Control Programme in Ireland.

    Science.gov (United States)

    McAloon, Conor G; Doherty, Michael L; Whyte, Paul; O'Grady, Luke; More, Simon J; Messam, Locksley L McV; Good, Margaret; Mullowney, Peter; Strain, Sam; Green, Martin J

    2016-06-01

    Bovine paratuberculosis is a disease characterised by chronic granulomatous enteritis which manifests clinically as a protein-losing enteropathy causing diarrhoea, hypoproteinaemia, emaciation and, eventually death. Some evidence exists to suggest a possible zoonotic link and a national voluntary Johne's Disease Control Programme was initiated by Animal Health Ireland in 2013. The objective of this study was to estimate herd-level true prevalence (HTP) and animal-level true prevalence (ATP) of paratuberculosis in Irish herds enrolled in the national voluntary JD control programme during 2013-14. Two datasets were used in this study. The first dataset had been collected in Ireland during 2005 (5822 animals from 119 herds), and was used to construct model priors. Model priors were updated with a primary (2013-14) dataset which included test records from 99,101 animals in 1039 dairy herds and was generated as part of the national voluntary JD control programme. The posterior estimate of HTP from the final Bayesian model was 0.23-0.34 with a 95% probability. Across all herds, the median ATP was found to be 0.032 (0.009, 0.145). This study represents the first use of Bayesian methodology to estimate the prevalence of paratuberculosis in Irish dairy herds. The HTP estimate was higher than previous Irish estimates but still lower than estimates from other major dairy producing countries. PMID:27237395

  6. Rapid and accurate identification of Streptococcus equi subspecies by MALDI-TOF MS

    DEFF Research Database (Denmark)

    Kudirkiene, Egle; Welker, Martin; Knudsen, Nanna Reumert;

    2015-01-01

    Streptococcus equi includes very important animal and human pathogens. S. equi subsp. equi (SEE) is a highly pathogenic equine specific subspecies, while S. equi subsp. zooepidemicus (SEZ) and S. equi subsp. ruminatorum are opportunistic pathogens of various animal species and humans. Due to great...... phenotypic and sequence similarity between three subspecies their discrimination remains difficult. In this study, we aimed to design and validate a novel, Superspectra based, MALDI-TOF MS approach for reliable, rapid and cost-effective identification of SEE and SEZ, the most frequent S. equi subspecies in...

  7. A distinctive new subspecies of Scytalopus griseicollis (Aves, Passeriformes, Rhinocryptidae from the northern Eastern Cordillera of Colombia and Venezuela

    Directory of Open Access Journals (Sweden)

    Jorge Avendaño

    2015-06-01

    Full Text Available We describe a new subspecies of Pale-bellied Tapaculo Scytalopus griseicollis from the northern Eastern Cordillera of Colombia and Venezuela. This form differs diagnosably in plumage from described subspecies S. g. griseicollis and S. g. gilesi and from the latter in tail length. It is also differentiated non-diagnosably in voice from both these populations. Ecological niche modelling analysis suggests that the new subspecies is restricted to the Andean montane forest and páramo north of both the arid Chicamocha valley and the Sierra Nevada del Cocuy.

  8. A distinctive new subspecies of Scytalopusgriseicollis (Aves, Passeriformes, Rhinocryptidae) from the northern Eastern Cordillera of Colombia and Venezuela.

    Science.gov (United States)

    Avendaño, Jorge Enrique; Donegan, Thomas M

    2015-01-01

    We describe a new subspecies of Pale-bellied Tapaculo Scytalopusgriseicollis from the northern Eastern Cordillera of Colombia and Venezuela. This form differs diagnosably in plumage from described subspecies Scytalopusgriseicollisgriseicollis and Scytalopusgriseicollisgilesi and from the latter in tail length. It is also differentiated non-diagnosably in voice from both these populations. Ecological niche modelling analysis suggests that the new subspecies is restricted to the Andean montane forest and páramo north of both the arid Chicamocha valley and the Sierra Nevada del Cocuy. PMID:26085800

  9. Inhibition of Tumor Necrosis Factor Alpha Alters Resistance to Mycobacterium avium Complex Infection in Mice

    OpenAIRE

    Bala, Shukal; Hastings, Kenneth L.; Kazempour, Kazem; Inglis, Shelly; Dempsey, Walla L.

    1998-01-01

    Increased production of tumor necrosis factor alpha (TNF-α) appears to play an important role in the progression of human immunodeficiency virus disease. One treatment strategy being explored is the use of TNF-α inhibitors. TNF-α also appears to be important in conferring resistance to infections, and the inhibition of this cytokine may exacerbate the emergence of opportunistic pathogens, such as Mycobacterium avium complex (MAC). The present study examines the possibility that inhibition of ...

  10. Mycobacterium avium-intracellulare brain abscess in HIV-positive patient

    OpenAIRE

    Sampada S Karne; Shashikala A Sangle; Dilip S Kiyawat; Dharmashale, Sujata N.; Kadam, Dilip B.; Bhardwaj, Renu S.

    2012-01-01

    Mycobacterial opportunistic infections are a major cause of morbidity and mortality among patients living with HIV (PLHIV) worldwide. Nontuberculous mycobacterial (NTM) infection is one of the leading causes of opportunistic infection in patients with advanced acquired immunodeficiency syndrome i.e., with CD4 count less than 50/cu.mm. Mycobacterium avium complex (MAC) is among the most common opportunistic bacterial infections in those patients with advanced immunodeficiency apart from crypto...

  11. In vitro and in vivo activities of clarithromycin against Mycobacterium avium.

    OpenAIRE

    Fernandes, P B; Hardy, D J; McDaniel, D; Hanson, C W; Swanson, R N

    1989-01-01

    There is no effective therapy to treat Mycobacterium avium complex infection in patients with acquired immune deficiency syndrome. Clarithromycin (A-56268; TE-031) is a new macrolide which is twofold more active than erythromycin against most aerobic bacteria. In addition, higher levels in serum and tissue are achieved with clarithromycin than with erythromycin. In this study, clarithromycin, erythromycin, difloxacin, temafloxacin, ciprofloxacin, rifampin, amikacin, and ethambutol were tested...

  12. Granuloma necrosis during Mycobacterium avium infection does not require tumor necrosis factor

    OpenAIRE

    Florido, M; Appelberg, R.

    2004-01-01

    The infection of tumor necrosis factor (TNF)-deficient mice with low doses of the virulent Mycobacterium avium strain 25291 led to the appearance of necrotic granulomas at 93 days of infection, i.e., sooner than necrotic granulomas appeared in C57BL/6 animals. Additionally, TNF-deficient mice exhibited higher mycobacterial loads in the infected organs, had extremely exacerbated gamma interferon responses as evaluated in the sera of infected animals, and showed reduced survival. Thus, TNF is n...

  13. Cathepsin L maturation and activity is impaired in macrophages harboring M. avium and M. tuberculosis.

    Science.gov (United States)

    Nepal, Rajeev M; Mampe, Stephanie; Shaffer, Brian; Erickson, Ann H; Bryant, Paula

    2006-06-01

    Mycobacterium tuberculosis-infected macrophages demonstrate diminished capacity to present antigens via class II MHC molecules. Since successful class II MHC-restricted antigen presentation relies on the actions of endocytic proteases, we asked whether the activities of cathepsins (Cat) B, S and L-three major lysosomal cysteine proteases-are modulated in macrophages infected with pathogenic Mycobacterium spp. Infection of murine bone marrow-derived macrophages with either Mycobacterium avium or M. tuberculosis had no obvious effect on Cat B or Cat S activity. In contrast, the activity of Cat L was altered in infected cells. Specifically, whereas the 24-kDa two-chain mature form of active Cat L predominated in uninfected cells, we observed an increase in the steady-state activity of the precursor single-chain (30 kDa) and 25-kDa two-chain forms of the enzyme in cells infected with either M. avium or M. tuberculosis. Pulse-chase analyses revealed that maturation of nascent, single-chain Cat L into the 25-kDa two-chain form was impaired in infected macrophages, and that maturation into the 24-kDa two-chain form did not occur. Consistent with these data, M. avium infection inhibited the IFNgamma-induced secretion of active two-chain Cat L by macrophages. Viable bacilli were not required to disrupt Cat L maturation, suggesting that a constitutively expressed mycobacterial component was responsible. The absence of the major active form of lysosomal Cat L in M. avium- and M. tuberculosis-infected macrophages may influence the types of T cell epitopes generated in these antigen-presenting cells, and/or the rate of class II MHC peptide loading. PMID:16636015

  14. Arbuscular mycorrhiza technology applied to micropropagated Prunus avium and to protection against Phytophthora cinnamomi

    OpenAIRE

    Cordier, Christelle; Trouvelot, A; GIANINAZZI, Silvio; Gianinazzi-Pearson, Vivienne

    1996-01-01

    Deux champignons endomycorhizogenes a arbuscules (Glomus intraradices, Glomus caledonium), introduits pendant la periode d'acclimatation, ont ete testes pour leur pouvoir infectieux et leur effet sur la croissance de plants micropropages de merisiers (Prunus avium L) transplantes dans deux differents sols neutres, prealablement desinfectes ou non. Les vitroplants endomycorhizes presentent une meilleure croissance que les plantes temoins sur les deux sols desinfectes. L'importance de l'augment...

  15. Isolation of Mycobacterium avium complex from water in the United States, Finland, Zaire, and Kenya.

    OpenAIRE

    von Reyn, C F; Waddell, R D; Eaton, T.; Arbeit, R D; Maslow, J.N.; Barber, T W; Brindle, R. J.; Gilks, C F; Lumio, J; Lähdevirta, J

    1993-01-01

    Disseminated infection with organisms of the Mycobacterium avium complex (MAC) is a common complication of AIDS in the United States and other developing countries, but it is rare or absent in sub-Saharan Africa. To assess the comparative likelihood of exposure to MAC in these geographic areas, we used a standard protocol to culture 91 water samples from environmental sites and piped water supply systems in the United States, Finland, Zaire, and Kenya. MAC was isolated from all geographic are...

  16. Comparative genomic characterization of Francisella tularensis strains belonging to low and high virulence subspecies.

    Directory of Open Access Journals (Sweden)

    Mia D Champion

    2009-05-01

    Full Text Available Tularemia is a geographically widespread, severely debilitating, and occasionally lethal disease in humans. It is caused by infection by a gram-negative bacterium, Francisella tularensis. In order to better understand its potency as an etiological agent as well as its potential as a biological weapon, we have completed draft assemblies and report the first complete genomic characterization of five strains belonging to the following different Francisella subspecies (subsp.: the F. tularensis subsp. tularensis FSC033, F. tularensis subsp. holarctica FSC257 and FSC022, and F. tularensis subsp. novicida GA99-3548 and GA99-3549 strains. Here, we report the sequencing of these strains and comparative genomic analysis with recently available public Francisella sequences, including the rare F. tularensis subsp. mediasiatica FSC147 strain isolate from the Central Asian Region. We report evidence for the occurrence of large-scale rearrangement events in strains of the holarctica subspecies, supporting previous proposals that further phylogenetic subdivisions of the Type B clade are likely. We also find a significant enrichment of disrupted or absent ORFs proximal to predicted breakpoints in the FSC022 strain, including a genetic component of the Type I restriction-modification defense system. Many of the pseudogenes identified are also disrupted in the closely related rarely human pathogenic F. tularensis subsp. mediasiatica FSC147 strain, including modulator of drug activity B (mdaB (FTT0961, which encodes a known NADPH quinone reductase involved in oxidative stress resistance. We have also identified genes exhibiting sequence similarity to effectors of the Type III (T3SS and components of the Type IV secretion systems (T4SS. One of the genes, msrA2 (FTT1797c, is disrupted in F. tularensis subsp. mediasiatica and has recently been shown to mediate bacterial pathogen survival in host organisms. Our findings suggest that in addition to the duplication of

  17. Complete Genome Sequence of Mycobacterium avium, Isolated from Commercial Domestic Pekin Ducks (Anas platyrhynchos domestica), Determined Using PacBio Single-Molecule Real-Time Technology.

    Science.gov (United States)

    Song, Xiao-Heng; Chen, Hong-Xi; Zhou, Wang-Shu; Wang, Jiang-Bo; Liu, Ma-Feng; Wang, Ming-Shu; Cheng, An-Chun; Jia, Ren-Yong; Chen, Shun; Sun, Kun-Feng; Yang, Qiao; Wu, Ying; Chen, Xiao-Yue; Zhu, De-Kang

    2016-01-01

    Mycobacterium avium is an important pathogenic bacterium in birds and has never, to our knowledge, reported to be isolated from domestic ducks. We present here the complete genome sequence of a virulent strain of Mycobacterium avium, isolated from domestic Pekin ducks for the first time, which was determined by PacBio single-molecule real-time technology. PMID:27587804

  18. Utility of morphological and molecular techniques for determination of paternity in two subspecies of Diabrotica undecimpunctata (Coleoptera: Chrysomelidae)

    Science.gov (United States)

    An experiment was conducted to determine the paternity of F1 progeny using morphological and molecular methods in Diabrotica (Coleoptera: Chrysomelidae) subspecies: Diabrotica undecimpunctata howardi Barber, also known as spotted cucumber beetle and D. undecimpunctata undecimpunctata Mannerheim, als...

  19. Urinary volatile molecules vary in males of the 2 European subspecies of the house mouse and their hybrids.

    Science.gov (United States)

    Mucignat-Caretta, C; Redaelli, M; Orsetti, A; Perriat-Sanguinet, M; Zagotto, G; Ganem, G

    2010-10-01

    Mice recognize other mice by identifying chemicals that confer a molecular signature to urinary marks. Such molecules may be involved in species recognition, and previous behavioral studies have related divergence of sexual preference between 2 subspecies of the house mouse (Mus musculus musculus and Mus musculus domesticus) to urinary odors. To characterize the differences between odors of males of the 2 subspecies and their first-generation offspring, the urinary volatile molecules were examined via gas chromatography coupled to mass spectrometry. Seven molecules were present in the samples from mice of at least one group. Their quantity varied among groups: M. m. domesticus showed a quantitatively richer panel of odorants in their urine when compared with M. m. musculus. The hybrids showed a more complex picture that was not directly related to one or the other parental subspecies. These quantitative differences may contribute to the specificity of the odorant bouquet of the 2 subspecies. PMID:20530376

  20. Yaws: 110 years after Castellani's discovery of Treponema pallidum subspecies pertenue.

    Science.gov (United States)

    Stamm, Lola V

    2015-07-01

    Yaws is a neglected infectious disease that affects mostly children and adolescents living in poor, rural communities in humid, tropical areas of Africa, southeast Asia, and the Pacific Islands. The etiological agent of yaws, Treponema pallidum subspecies pertenue (T. pertenue), was discovered by Aldo Castellani in 1905 shortly after Schaudinn and Hoffmann discovered the etiological agent of syphilis, T. pallidum subspecies pallidum. The discovery of T. pertenue enabled the development of animal models and the identification of an effective antibiotic treatment (i.e., penicillin) for yaws. A World Health Organization (WHO) mass treatment campaign from 1952 to 1964 reduced the global burden of yaws by 95%, but failed to eradicate this disease. Today, 110 years after Castellani's discovery of T. pertenue, yaws is again targeted for eradication. Recent advances in the treatment and diagnosis of yaws improve the likelihood of success this time. However, several challenges must be overcome to make the goal of yaws eradication attainable. PMID:25870417

  1. Evaluation of a PCR assay for identification and differentiation of Campylobacter fetus subspecies

    DEFF Research Database (Denmark)

    Hum, S.; Quinn, K.; Brunner, J.; On, Stephen L.W.

    1997-01-01

    Objective To evaluate a polymerase chain reaction assay for identification of Campylobacter fetus and differentiation of the defined subspecies. Design Characterisation of bacterial strains by traditional phenotyping, polymerase chain reaction, a probabilistic identification scheme and...... macrorestriction profiling using pulsed field gel electrophoresis. Procedure The results of identification of 99 bacterial strains as determined by conventional phenotyping or by polymerase chain reaction were compared. Two of these were type strains of C fetus subsp fetus and C fetus subsp venerealis; the...... remaining strains were field isolates putatively identified as C fetus. In cases where the subspecies identity was disputed, isolates were identified by means of a probabilistic identification scheme and by macrorestriction profiling. Results The agreement between strain identities initially suggested by...

  2. Redescription of Gammarus pseudosyriacus (Karaman & Pinkster, 1977) and description of a new subspecies from southern Iran (Crustacea, Amphipoda, Gammaridae)

    OpenAIRE

    Semsar-kazerooni,Maryam; Zamanpoore, Mehrdad; Sadeghi, Saber

    2016-01-01

    The present study focused on redescription of Gammarus pseudosyriacus (Karaman & Pinkster, 1977) based on new materials from Zagros Mountains and describes a new subspecies of freshwater amphipod, Gammarus pseudosyriacus issatisi subsp. n., from the southern Zagros Mountains. The work is based on morphological and morphometric comparisons. This new subspecies has features similar to Gammarus pseudosyriacus. The distinct features that distinguish Gammarus pseudosyriacus issatisi subsp. n. ...

  3. Isolation of Multiple Subspecies of Bacillus thuringiensis from a Population of the European Sunflower Moth, Homoeosoma nebulella

    OpenAIRE

    Itoua-Apoyolo, C.; Drif, L.; Vassal, J. M.; DeBarjac, H.; Bossy, J. P.; Leclant, F.; Frutos, R

    1995-01-01

    Five subspecies of Bacillus thuringiensis were isolated from dead and diseased larvae obtained from a laboratory colony of the European sunflower moth, Homoeosoma nebulella. The subspecies isolated were B. thuringiensis subspp. thuringiensis (H 1a), kurstaki (H 3a3b3c), aizawai (H 7), morrisoni (H 8a8b), and thompsoni (H 12). Most isolates produced typical bipyramidal crystals, but the B. thuringiensis subsp. thuringiensis isolate produced spherical crystals and the B. thuringiensis subsp. th...

  4. Molecular typing of Salmonella enterica subspecies enterica serovar Typhimurium isolated in Abruzzo region (Italy) from 2008 to 2010

    OpenAIRE

    Alessandra Alessiani; Lorena Sacchini; Eugenio Pontieri; Jacopo Gavini; Elisabetta Di Giannatale

    2014-01-01

    In this study, 47 antibiotic-resistant strains of Salmonella enterica subspecies enterica serovar Typhimurium (ST) were characterised, including 15 monophasic variants 1, 4, [5], 12:i:-, (STm) isolated from different matrices. They were all selected from 389 Salmonella enterica subspecies enterica strains isolated during 2008-2010 in Abruzzo region (Italy). Thirty-seven strains showed to be resistant to more than 1 antibiotic. Among 47 isolates, phage type U311 and DT104 were identified. The ...

  5. Genetic structure and subspecies divergence of wild boars Sus scrofa in mainland China based on the microsatellite variation analysis

    OpenAIRE

    ZHANG Bao-Wei; ZHANG Chen-Ling; CHEN Jian-Qin; Ding, Dong; LI Chong-Qi; Zhou, Kai-Ya; Chang, Qing

    2008-01-01

    In order to discuss the subspecies classification of wild boar Sus scrofa in China, We studied their genetic structure using 11 microsatellites, with emphasis on South China. High levels of genetic variationwere detected in South China (SC), North China (NC) and Northeast China (NE) wild boar populations. This was consistent with the huge population size of the wild boar and its status as a widely-distributed species. Weak genetic divergence was observed in all subspecies involved in this s...

  6. A new attempt to distinguish serologically the subspecies of Treponema pallidum causing syphilis and yaws.

    OpenAIRE

    Noordhoek, G T; Cockayne, A; Schouls, L. M.; Meloen, R H; Stolz, E; van Embden, J D

    1990-01-01

    In an effort to serologically differentiate syphilis from yaws, 69 monoclonal antibody species raised against Treponema pallidum subsp. pallidum were tested by immunoblotting for their reactivity with Treponema pallidum subsp. pertenue. All monoclonal antibodies reacted with antigens with the same molecular weight of both subspecies. Furthermore, no differences in reactivity between sera from yaws patients and from syphilis patients were found by Western blot (immunoblot) analysis of cell lys...

  7. A new subspecies of Jerboa from Turkey; Allactaga euphratica kivanci subsp. n.

    OpenAIRE

    ÇOLAK, Ercüment; Nuri YİĞİT

    1998-01-01

    In this study, 42 specimens of Allactaga euphratica from Urfa (Turkey) were examined based on morphological, cranial, external and karyological characters. A comparison of specimens from Urfa with those from Syria, Jordan, Kuwait and Iraq showed that specimens from Urfa, Syria and Jordan, Kuwait and Iraq showed that specimens from Urfa, Syria and Jordan differ from those from Kuwait and Iraq. Thus, specimens from Urfa (Turkey), Syria and Jordan were included in a new subspecies, Allactaga ...

  8. Characterization of new IS elements and studies of their dispersion in two subspecies of Leifsonia xyli

    OpenAIRE

    Monteiro-Vitorello Claudia B.; Camargo Luis; Van Sluys Marie-Anne; Zerillo Marcelo M

    2008-01-01

    Abstract Background Leifsonia xyli is a xylem-inhabiting bacterial species comprised of two subspecies: L. xyli subsp. xyli (Lxx) and L. xyli subsp. cynodontis (Lxc). Lxx is the causal agent of ratoon stunting disease in sugarcane commercial fields and Lxc colonizes the xylem of several grasses causing either mild or no symptoms of disease. The completely sequenced genome of Lxx provided insights into its biology and pathogenicity. Since IS elements are largely reported as an important source...

  9. Essential Oil Compositions of Subspecies of Scutellaria Brevibracteata Stapf from Turkey

    Directory of Open Access Journals (Sweden)

    M. ÇİÇEK

    2014-06-01

    Full Text Available Scutellaria L., with nearly 400 species, is one of the largest genera of the family Lamiaceae. In the last taxonomic treatment conducted by the first author, the genus Scutellaria has been evaluated as a genus with 33 taxa consisting of 25 species, 1 hybrid species and 13 subspecies in the Flora of Turkey. Turkish Scutellaria species are classified under three sections: section Scutellaria, section Salviifoliae and section Lupulinaria. We have here studied essential oil composition of the subspecies of S. brevibracteata belonging to the section Scutellaria. In the Anatolian traditional medicine, there is no more information on usage of Scutellaria except for the usage for constipation, hemostatic, tonic and wound healing, due to lack of its aromatic odour.Dried the aerial parts of three subspecies of Scutellaria brevibracteata Stapf from Turkey were hydrodistilled. The chemical compositions of the essential oils obtained by hydrodistillation were analyzed by gas chromatography (GC and gas chromatography/mass spectrometry (GC/MS, simultaneously. The subspecies of S. brevibracteata investigated here as follows: S. brevibractetata Stapf subsp. brevibracteata, S. brevibractetata Stapf subsp. subvelutina (Rech.f. Greuter & Burdet and S. brevibractetata Stapf subsp. pannosula (Rech.f. Greuter & Burdet.According to the results of analyzing, β-caryophyllene (22.8% and caryophyllene oxide (16.0% were determined as the main components in S. brevibractetata subsp. brevibracteata oil. In S. brevibractetata subsp. subvelutina oil, β-caryophyllene (28.3%, linalool (12.4%, hexadecanoic acid (10.8% were found as major constituents. β-caryophyllene (36.4%, a-cadinol (9.8%, d-cadinene (7.0% linalool (5.3% were identified as major component in the oils of S. brevibractetata subsp. pannosula.

  10. Essential Oil Compositions of Subspecies of Scutellaria Brevibracteata Stapf from Turkey

    OpenAIRE

    M. ÇİÇEK; Yilmaz, G.; B. DEMIRCI; K. H.C. BAŞER

    2014-01-01

    Scutellaria L., with nearly 400 species, is one of the largest genera of the family Lamiaceae. In the last taxonomic treatment conducted by the first author, the genus Scutellaria has been evaluated as a genus with 33 taxa consisting of 25 species, 1 hybrid species and 13 subspecies in the Flora of Turkey. Turkish Scutellaria species are classified under three sections: section Scutellaria, section Salviifoliae and section Lupulinaria. We have here studied essential oil composition of the sub...

  11. Presence and distribution of two sub-species of Eurema agave (Lepidoptera, Pieridae in Costa Rica

    Directory of Open Access Journals (Sweden)

    Jim Cordoba-Alfaro

    2011-06-01

    Full Text Available Austin (1992 reported Eurema a. agave (Cramer 1775 to the Caribbean of Costa Rica. However, he actually had found E. a. millerorum, described by Bousquets & Luis-Martinez (1987 for the Caribbean of Mexico. The presence of Eurema a. agave is confirmed on this paper with information of specimens collected in the Pacific and Atlantic slopes of Costa Rica. Aspects on distribution of both subspecies are included.

  12. Seasonal and geographic variation of southern blue whale subspecies in the Indian ocean.

    OpenAIRE

    Flore Samaran; Stafford, Kathleen M.; Branch, Trevor A.; Jason Gedamke; Jean-Yves Royer; Dziak, Robert P.; Christophe Guinet

    2013-01-01

    International audience Understanding the seasonal movements and distribution patterns of migratory species over ocean basin scales is vital for appropriate conservation and management measures. However, assessing populations over remote regions is challenging, particularly if they are rare. Blue whales (Balaenoptera musculus spp) are an endangered species found in the Southern and Indian Oceans. Here two recognized subspecies of blue whales and, based on passive acoustic monitoring, four "...

  13. Presence and distribution of two sub-species of Eurema agave (Lepidoptera, Pieridae) in Costa Rica

    OpenAIRE

    Jim Cordoba-Alfaro; Luis Ricardo Murillo-Hiller

    2011-01-01

    Austin (1992) reported Eurema a. agave (Cramer 1775) to the Caribbean of Costa Rica. However, he actually had found E. a. millerorum, described by Bousquets & Luis-Martinez (1987) for the Caribbean of Mexico. The presence of Eurema a. agave is confirmed on this paper with information of specimens collected in the Pacific and Atlantic slopes of Costa Rica. Aspects on distribution of both subspecies are included.

  14. Novel Antibacterial Activity of Lactococcus Lactis Subspecies Lactis Z11 Isolated from Zabady

    OpenAIRE

    Enan, Gamal; Abdel-Shafi, Seham; Ouda, Sahar; Negm, Sally

    2013-01-01

    The purpose of this study was to select and characterize a probiotic bacterium with distinctive antimicrobial activities. In this respect, Lactococcus lactis subspecies lactis Z11 (L. lactis Z11) isolated from Zabady (Arabian yoghurt) inhibited other strains of lactic acid bacteria and some food-born pathogens including Listeria monocytogenes, Bacillus cereus and staphylococcus aureus. The inhibitory activity of cell free supernatant (CFS) of L. lactis Z11 isolated from zabady was lost by pro...

  15. Inference of Purifying and Positive Selection in Three Subspecies of Chimpanzees (Pan troglodytes) from Exome Sequencing

    OpenAIRE

    Bataillon, Thomas; Duan, Jinjie; Hvilsom, Christina; Jin, Xin; Li, Yingrui; Skov, Laurits; Glemin, Sylvain; Munch, Kasper; Jiang, Tao; Qian, Yu; Hobolth, Asger; Wang, Jun; Mailund, Thomas; Siegismund, Hans R.; Schierup, Mikkel H.

    2015-01-01

    We study genome-wide nucleotide diversity in three subspecies of extant chimpanzees using exome capture. After strict filtering, Single Nucleotide Polymorphisms and indels were called and genotyped for greater than 50% of exons at a mean coverage of 35× per individual. Central chimpanzees (Pan troglodytes troglodytes) are the most polymorphic (nucleotide diversity, θw = 0.0023 per site) followed by Eastern (P. t. schweinfurthii) chimpanzees (θw = 0.0016) and Western (P. t. verus) chimpanzees ...

  16. EFFECT OF PHENOMENON XENIA ON TRAIT SEED OF MAIZE SUB-SPECIES CROSSES

    OpenAIRE

    Mohammed Dhary Yousif EL-JUBOURI

    2012-01-01

    To investigate the effect of phenomenon xenia in maize seeds, five genotypes were intercrossed. One genotype of each of (Zea mays L.) sub-species (Everta, Indurata, saccharata, Indentata and the white endosperm Denprofeski were crossbred. F1 and F2 seeds were investigated .The results showed that hybrid vigor or dominance were controlling seed trait. The gene (Y) of yellow kernel was dominant over white (y) with possibility of modifying gene sharing action on this trait. Reciprocal gave 1:3 r...

  17. Characterization of Vitis vinifera L. subspecies sylvestris (Gmelin) Hegi in the Ebro river Basin (Spain)

    OpenAIRE

    Ocete Rubio, Rafael; Arroyo García, R.; Morales, M. L.; Cantos, Manuel; Gallardo Cano, A.; Pérez Izquierdo, Mª Á.; Gómez Parrales, Isidoro; López Martínez, Mª Ángeles

    2011-01-01

    A wild grapevine population of 76 vines of Vitis vinifera L. subspecies sylvestris (Gmelin) Hegi was found along the Iregua river valley (Northeastern Spain), located in the Ibérica mountain-range. The characterization of this phytogenetic resource was based on an ampelographic description of the male and female individuals and an evaluation of their sanitary state. This shows that the imported North American downy and powdery mildews are the main pathogens, but no symptoms caused by phylloxe...

  18. Biological, ecological, conservation and legal information for all species and subspecies of Australian bird

    OpenAIRE

    Garnett, Stephen T.; Duursma, Daisy E.; Ehmke, Glenn; Guay, Patrick-Jean; Stewart, Alistair; Szabo, Judit K.; Weston, Michael A; Bennett, Simon; Crowley, Gabriel M.; Drynan, David; Dutson, Guy; Fitzherbert, Kate; Donald C Franklin

    2015-01-01

    We introduce a dataset of biological, ecological, conservation and legal information for every species and subspecies of Australian bird, 2056 taxa or populations in total. Version 1 contains 230 fields grouped under the following headings: Taxonomy & nomenclature, Phylogeny, Australian population status, Conservation status, Legal status, Distribution, Morphology, Habitat, Food, Behaviour, Breeding, Mobility and Climate metrics. It is envisaged that the dataset will be updated periodically w...

  19. Bifidobacterium animalis subsp. lactis ATCC 27673 Is a Genomically Unique Strain within Its Conserved Subspecies

    OpenAIRE

    Loquasto, Joseph R.; Barrangou, Rodolphe; Dudley, Edward G.; Stahl, Buffy; Chen, Chun; Roberts, Robert F.

    2013-01-01

    Many strains of Bifidobacterium animalis subsp. lactis are considered health-promoting probiotic microorganisms and are commonly formulated into fermented dairy foods. Analyses of previously sequenced genomes of B. animalis subsp. lactis have revealed little genetic diversity, suggesting that it is a monomorphic subspecies. However, during a multilocus sequence typing survey of Bifidobacterium, it was revealed that B. animalis subsp. lactis ATCC 27673 gave a profile distinct from that of the ...

  20. Fine Analysis of Genetic Diversity of the tpr Gene Family among Treponemal Species, Subspecies and Strains

    OpenAIRE

    Centurion-Lara, Arturo; Giacani, Lorenzo; Godornes, Charmie; Molini, Barbara J.; Brinck Reid, Tara; Lukehart, Sheila A.

    2013-01-01

    Background The pathogenic non-cultivable treponemes include three subspecies of Treponema pallidum (pallidum, pertenue, endemicum), T. carateum, T. paraluiscuniculi, and the unclassified Fribourg-Blanc treponeme (Simian isolate). These treponemes are morphologically indistinguishable and antigenically and genetically highly similar, yet cross-immunity is variable or non-existent. Although all of these organisms cause chronic, multistage skin and systemic disease, they have historically been c...

  1. A new subspecies of Chamaea fasciata (Wrentit) from Oregon (Aves: Timaliinae)

    Science.gov (United States)

    Browning, M. Ralph

    1992-01-01

    Geographic variation in plumage color of Chamaea fasciata (Wrentit) from northern California and southern Oregon is related to climate. A new subspecies, Chamaea fasciata margra, is described from a disjunct population of southern interior Oregon. Colonization of C. fasciata in interior Oregon was perhaps from birds crossing coniferous forests via isolated balds of Ceonothus. Recent increases of Wrentits in interior Oregon may be in response to habitat alterations (deforestation, fires) and concurrent global warming.

  2. A morphological review of subspecies of the Asian box turtle, Cuora amboinensis (Testudines, Geomydidae)

    Science.gov (United States)

    Ernst, Carl H.; Laemmerzahl, Arndt F.; Lovich, Jeffrey E.

    2016-01-01

    The turtle Cuora amboinensis has an extensive distribution covering most of southern mainland Asia, Indonesia, and extending to the Philippine Islands. Unlike many species, C. amboinensis occurs on both sides of Wallace's Line separating Asian and Australian flora and fauna. Four subspecies are currently recognized; Cuora a. kamaroma (southern continental Asia, Java and the northern Philippines [introduced]), C. a. lineata (Kachin Province, Myanmar [Burma] and adjacent Yunnan Province, China), C. a. couro (Sumatra, Java, Sumbawa, and adjacent smaller Indonesian islands); and C. a. amboinensis (Moluccas, Sulawesi, Philippines). Five pattern and 33 morphological characters were examined for variation in 691 individuals from throughout the species' range. Our analyses suggest that only two presently recognized subspecies are valid: amboinensis andkamaroma. Neither couro nor lineata are supported by our analysis. We recommend that C. a. couroshould be synonymized with the species C. amboinensis and C. a. lineata with the subspecies C. a. kamaroma.

  3. Chimpanzee subspecies and ‘robust’ australopithecine holotypes, in the context of comments by Darwin

    Directory of Open Access Journals (Sweden)

    S. Prat

    2010-02-01

    Full Text Available On the basis of comparative anatomy (including chimpanzees, gorillas and other primates, Darwin1 suggested that Africa was the continent from which ‘progenitors’ of humankind evolved. Hominin fossils from this continent proved him correct. We present the results of morphometric analyses based on cranial data obtained from chimpanzee taxa currently recognised as distinct subspecies, namely Pan troglodytes troglodytes and Pan troglodytes schweinfurthii, as well as Pan paniscus (bonobo. Our objective was to use a morphometric technique2 to quantify the degree of similarity between pairs of specimens, in the context of a statistical (probabilistic definition of a species.3–5 Results obtained from great apes, including two subspecies of chimpanzee, were assessed in relation to same-scale comparisons between the holotypes of ‘robust’ australopithecine (Plio-Pleistocene hominin taxa which have traditionally been distinguished at a species level, notably Paranthropus robustus from South Africa, and Paranthropus (Australopithecus/ Zinjanthropus boisei from East Africa. The question arises as to whether the holotypes of these two taxa, TM 1517 from Kromdraai6 and OH 5 from Olduvai,7 respectively, are different at the subspecies rather than at the species level.

  4. Ecotype evolution in Glossina palpalis subspecies, major vectors of sleeping sickness.

    Directory of Open Access Journals (Sweden)

    Thierry De Meeûs

    2015-03-01

    Full Text Available BACKGROUND: The role of environmental factors in driving adaptive trajectories of living organisms is still being debated. This is even more important to understand when dealing with important neglected diseases and their vectors. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we analysed genetic divergence, computed from seven microsatellite loci, of 614 tsetse flies (Glossina palpalis gambiensis and Glossina palpalis palpalis, major vectors of animal and human trypanosomes from 28 sites of West and Central Africa. We found that the two subspecies are so divergent that they deserve the species status. Controlling for geographic and time distances that separate these samples, which have a significant effect, we found that G. p. gambiensis from different landscapes (Niayes of Senegal, savannah and coastal environments were significantly genetically different and thus represent different ecotypes or subspecies. We also confirm that G. p. palpalis from Ivory Coast, Cameroon and DRC are strongly divergent. CONCLUSIONS/SIGNIFICANCE: These results provide an opportunity to examine whether new tsetse fly ecotypes might display different behaviour, dispersal patterns, host preferences and vectorial capacities. This work also urges a revision of taxonomic status of Glossina palpalis subspecies and highlights again how fast ecological divergence can be, especially in host-parasite-vector systems.

  5. Ecotype Evolution in Glossina palpalis Subspecies, Major Vectors of Sleeping Sickness

    Science.gov (United States)

    De Meeûs, Thierry; Bouyer, Jérémy; Ravel, Sophie; Solano, Philippe

    2015-01-01

    Background The role of environmental factors in driving adaptive trajectories of living organisms is still being debated. This is even more important to understand when dealing with important neglected diseases and their vectors. Methodology/Principal Findings In this paper, we analysed genetic divergence, computed from seven microsatellite loci, of 614 tsetse flies (Glossina palpalis gambiensis and Glossina palpalis palpalis, major vectors of animal and human trypanosomes) from 28 sites of West and Central Africa. We found that the two subspecies are so divergent that they deserve the species status. Controlling for geographic and time distances that separate these samples, which have a significant effect, we found that G. p. gambiensis from different landscapes (Niayes of Senegal, savannah and coastal environments) were significantly genetically different and thus represent different ecotypes or subspecies. We also confirm that G. p. palpalis from Ivory Coast, Cameroon and DRC are strongly divergent. Conclusions/Significance These results provide an opportunity to examine whether new tsetse fly ecotypes might display different behaviour, dispersal patterns, host preferences and vectorial capacities. This work also urges a revision of taxonomic status of Glossina palpalis subspecies and highlights again how fast ecological divergence can be, especially in host-parasite-vector systems. PMID:25775377

  6. Mitogenomic phylogenetics of fin whales (Balaenoptera physalus spp.: genetic evidence for revision of subspecies.

    Directory of Open Access Journals (Sweden)

    Frederick I Archer

    Full Text Available There are three described subspecies of fin whales (Balaenoptera physalus: B. p. physalus Linnaeus, 1758 in the Northern Hemisphere, B. p. quoyi Fischer, 1829 in the Southern Hemisphere, and a recently described pygmy form, B. p. patachonica Burmeister, 1865. The discrete distribution in the North Pacific and North Atlantic raises the question of whether a single Northern Hemisphere subspecies is valid. We assess phylogenetic patterns using ~16 K base pairs of the complete mitogenome for 154 fin whales from the North Pacific, North Atlantic--including the Mediterranean Sea--and Southern Hemisphere. A Bayesian tree of the resulting 136 haplotypes revealed several well-supported clades representing each ocean basin, with no haplotypes shared among ocean basins. The North Atlantic haplotypes (n = 12 form a sister clade to those from the Southern Hemisphere (n = 42. The estimated time to most recent common ancestor (TMRCA for this Atlantic/Southern Hemisphere clade and 81 of the 97 samples from the North Pacific was approximately 2 Ma. 14 of the remaining North Pacific samples formed a well-supported clade within the Southern Hemisphere. The TMRCA for this node suggests that at least one female from the Southern Hemisphere immigrated to the North Pacific approximately 0.37 Ma. These results provide strong evidence that North Pacific and North Atlantic fin whales should not be considered the same subspecies, and suggest the need for revision of the global taxonomy of the species.

  7. Evolutionary divergence of geographic subspecies within the scalloped spiny lobster Panulirus homarus (Linnaeus 1758.

    Directory of Open Access Journals (Sweden)

    Shane D Lavery

    Full Text Available Panulirus homarus is an economically important spiny lobster that is widespread through the Indo-West Pacific Region, but has an uncertain taxonomic status, with three or four geographic subspecies having been described. This study used mitochondrial (16S, COI and control region and nuclear (18S, ITS-1 DNA sequences to examine specimens of all putative subspecies and forms from throughout their range, in order to determine their genetic validity, and understand the evolutionary history of this species. Despite the range of diversity present in the loci examined, the results were consistent across genes. P. h. rubellus from the SW Indian Ocean comprised the most divergent lineage that was reciprocally monophyletic with respect to all other P. homarus (approx. 9% divergence in COI, and has likely evolved reproductive barriers. The putative P. h. "Brown" subspecies from the Marquesas Is in the central Pacific also comprised a somewhat divergent monophyletic lineage (approx. 3% in COI, but may simply be an allopatric population. The widespread P. h. homarus was not diverged at all from the described P. h. megasculpta from the NW Indian Ocean. The degree of evolutionary divergence of populations at the extremes distribution of the species is somewhat surprising, given the long pelagic larval stage, but suggests that allopatric speciation has been an important driver in the evolution of the genus.

  8. Building a DNA barcode reference library for the true butterflies (Lepidoptera of Peninsula Malaysia: what about the subspecies?

    Directory of Open Access Journals (Sweden)

    John-James Wilson

    Full Text Available The objective of this study was to build a DNA barcode reference library for the true butterflies of Peninsula Malaysia and assess the value of attaching subspecies names to DNA barcode records. A new DNA barcode library was constructed with butterflies from the Museum of Zoology, University of Malaya collection. The library was analysed in conjunction with publicly available DNA barcodes from other Asia-Pacific localities to test the ability of the DNA barcodes to discriminate species and subspecies. Analyses confirmed the capacity of the new DNA barcode reference library to distinguish the vast majority of species (92% and revealed that most subspecies possessed unique DNA barcodes (84%. In some cases conspecific subspecies exhibited genetic distances between their DNA barcodes that are typically seen between species, and these were often taxa that have previously been regarded as full species. Subspecies designations as shorthand for geographically and morphologically differentiated groups provide a useful heuristic for assessing how such groups correlate with clustering patterns of DNA barcodes, especially as the number of DNA barcodes per species in reference libraries increases. Our study demonstrates the value in attaching subspecies names to DNA barcode records as they can reveal a history of taxonomic concepts and expose important units of biodiversity.

  9. Building a DNA Barcode Reference Library for the True Butterflies (Lepidoptera) of Peninsula Malaysia: What about the Subspecies?

    Science.gov (United States)

    Wilson, John-James; Sing, Kong-Wah; Sofian-Azirun, Mohd

    2013-01-01

    The objective of this study was to build a DNA barcode reference library for the true butterflies of Peninsula Malaysia and assess the value of attaching subspecies names to DNA barcode records. A new DNA barcode library was constructed with butterflies from the Museum of Zoology, University of Malaya collection. The library was analysed in conjunction with publicly available DNA barcodes from other Asia-Pacific localities to test the ability of the DNA barcodes to discriminate species and subspecies. Analyses confirmed the capacity of the new DNA barcode reference library to distinguish the vast majority of species (92%) and revealed that most subspecies possessed unique DNA barcodes (84%). In some cases conspecific subspecies exhibited genetic distances between their DNA barcodes that are typically seen between species, and these were often taxa that have previously been regarded as full species. Subspecies designations as shorthand for geographically and morphologically differentiated groups provide a useful heuristic for assessing how such groups correlate with clustering patterns of DNA barcodes, especially as the number of DNA barcodes per species in reference libraries increases. Our study demonstrates the value in attaching subspecies names to DNA barcode records as they can reveal a history of taxonomic concepts and expose important units of biodiversity. PMID:24282514

  10. NCBI nr-aa BLAST: CBRC-XTRO-01-3068 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-XTRO-01-3068 ref|NP_960250.1| AppC [Mycobacterium avium subsp. paratuberculosis... K-10] gb|AAS03633.1| AppC [Mycobacterium avium subsp. paratuberculosis K-10] NP_960250.1 2.1 28% ...

  11. Proposed minimal standards for the description of genera, species and subspecies of the Pasteurellaceae.

    Science.gov (United States)

    Christensen, Henrik; Kuhnert, Peter; Busse, Hans-Jürgen; Frederiksen, Wilhelm C; Bisgaard, Magne

    2007-01-01

    , and at least two phenotypic differences from existing species should be identified if possible. We recommend the use of the subspecies category only for subgroups associated with disease or similar biological characteristics. At the subspecies level, the genotypic groups must always be nested within the boundaries of an existing species. Phenotypic cohesion must be documented at the subspecies level and separation between subspecies and related species must be fully documented, as well as association with particular disease and host. An overview of methods previously used to characterize isolates of the Pasteurellaceae has been given. Genotypic and phenotypic methods are separated in relation to tests for investigating diversity and cohesion and to separate taxa at the level of genus as well as species and subspecies. PMID:17220461

  12. Immuno-enhancement of Taishan Pinus massoniana pollen polysaccharides on recombinant Bordetella avium ompA expressed in Pichia pastoris.

    Science.gov (United States)

    Liu, Liping; Yu, Cuilian; Wang, Chuanwen; Shao, Mingxu; Yan, Zhengui; Jiang, Xiaodong; Chi, Shanshan; Wang, Zhen; Wei, Kai; Zhu, Ruiliang

    2016-06-01

    Bordetellosis, caused by Bordetella avium, continues to be an economic problem in the poultry industry of China. Vaccines with good protective ability are lacking. Thus, developing a novel vaccine against the B. avium infection is crucial. Here, we constructed a recombinant Pichia pastoris transformant capable of expressing the outer membrane protein A (ompA) of B. avium to prepare the recombinant ompA subunit vaccine and then evaluated its immune effects. To further investigate the immunomodulation effects of Taishan Pinus massoniana pollen polysaccharides (TPPPS) on this subunit vaccine, three concentrations (20, 40, and 60 mg/mL) of TPPPS were used as the adjuvants of the ompA subunit vaccine respectively. The conventional Freund's incomplete adjuvant served as the control of TPPPS. Chickens in different groups were separately vaccinated with these vaccines thrice. During the monitoring period, serum antibody titers, concentrations of serum IL-4, percentages of CD4(+) and CD8(+) T-lymphocytes in the peripheral blood, lymphocyte transformation rate, and protection rate were detected. Results showed that the pure ompA vaccine induced the production of anti-ompA antibody, the secretion of IL-4, the increase of CD4(+) T-lymphocytes counts and lymphocyte transformation rate in the peripheral blood. Moreover, the pure ompA vaccine provided a protection rate of 71.67% after the B. avium challenge. Notably, TPPPS adjuvant vaccines induced higher levels of immune responses than the pure ompA vaccine, and 60 mg/mL TPPPS adjuvant vaccine showed optimal immune effects and had a 91.67% protection rate. Our findings indicated that this recombinant B. avium ompA subunit vaccine combined with TPPPS had high immunostimulatory potential. Results provided a new perspective for B. avium subunit vaccine research. PMID:26975477

  13. Threatened and endangered subspecies with vulnerable ecological traits also have high susceptibility to sea level rise and habitat fragmentation.

    Science.gov (United States)

    Benscoter, Allison M; Reece, Joshua S; Noss, Reed F; Brandt, Laura A; Mazzotti, Frank J; Romañach, Stephanie S; Watling, James I

    2013-01-01

    The presence of multiple interacting threats to biodiversity and the increasing rate of species extinction make it critical to prioritize management efforts on species and communities that maximize conservation success. We implemented a multi-step approach that coupled vulnerability assessments evaluating threats to Florida taxa such as climate change, sea-level rise, and habitat fragmentation with in-depth literature surveys of taxon-specific ecological traits. The vulnerability, adaptive capacity, and ecological traits of 12 threatened and endangered subspecies were compared to non-listed subspecies of the same parent species. Overall, the threatened and endangered subspecies showed high vulnerability and low adaptive capacity, in particular to sea level rise and habitat fragmentation. They also exhibited larger home ranges and greater dispersal limitation compared to non-endangered subspecies, which may inhibit their ability to track changing climate in fragmented landscapes. There was evidence for lower reproductive capacity in some of the threatened or endangered taxa, but not for most. Taxa located in the Florida Keys or in other low coastal areas were most vulnerable to sea level rise, and also showed low levels of adaptive capacity, indicating they may have a lower probability of conservation success. Our analysis of at-risk subspecies and closely related non-endangered subspecies demonstrates that ecological traits help to explain observed differences in vulnerability and adaptive capacity. This study points to the importance of assessing the relative contributions of multiple threats and evaluating conservation value at the species (or subspecies) level when resources are limited and several factors affect conservation success. PMID:23940614

  14. Inactivation of Mycobacterium paratuberculosis and Mycobacterium tuberculosis in fresh soft cheese by gamma radiation

    Science.gov (United States)

    Badr, Hesham M.

    2011-11-01

    The effectiveness of gamma irradiation on the inactivation of Mycobacterium paratuberculosis, Mycobacterium bovis and Mycobacterium tuberculosis in fresh soft cheese that prepared from artificially inoculated milk samples was studied. Irradiation at dose of 2 kGy was sufficient for the complete inactivation of these mycobacteria as they were not detected in the treated samples during storage at 4±1 °C for 15 days. Moreover, irradiation of cheese samples, that were prepared from un-inoculated milk, at this effective dose had no significant effects on their gross composition and contents from riboflavin, niacin and pantothenic acid, while significant decreases in vitamin A and thiamin were observed. In addition, irradiation of cheese samples had no significant effects on their pH and nitrogen fractions contents, except for the contents of ammonia, which showed a slight, but significant, increases due to irradiation. The analysis of cheese fats indicated that irradiation treatment induced significant increase in their oxidation parameters and contents from free fatty acids; however, the observed increases were relatively low. On the other hand, irradiation of cheese samples induced no significant alterations on their sensory properties. Thus, irradiation dose of 2 kGy can be effectively applied to ensure the safety of soft cheese with regards to these harmful mycobacteria.

  15. Enfermedad pulmonar por especies del complejo Mycobacterium avium intracellulare y correspondencia terapéutica Mycobacterium avium intracellulare complex pulmonary disease species and therapeutic match

    Directory of Open Access Journals (Sweden)

    Hayvin Pérez Cruz

    2013-04-01

    Full Text Available Introducción: la preocupación mundial ante las enfermedades causadas por micobacterias ambientales es creciente, debido a las dificultades diagnósticas y de tratamiento. Objetivos: determinar el comportamiento de la enfermedad pulmonar por especies del complejo Mycobacterium avium intracellulare y la correspondencia con la terapéutica recibida en el Hospital Neumológico Benéfico Jurídico de La Habana, durante el período 2000-2010. Métodos: se realizó un estudio descriptivo retrospectivo, que abarcó 55 pacientes con el diagnóstico de la enfermedad, los cuales cumplieron con los criterios de inclusión-exclusión predefinidos; la tendencia en el registro de diagnósticos se basó en el coeficiente de correlación lineal; en las variables cualitativas y cuantitativas discretas se usaron distribuciones de frecuencia con el cálculo del porcentaje y su respectivo intervalo de confianza de 95 %. Resultados: se obtuvo una tendencia decreciente en el diagnóstico de enfermedad pulmonar por especies del complejo Mycobacterium avium intracellulare, el 47,3 % de los pacientes estudiados se encontraba entre los 50 y 69 años de edad. La mayor parte del grupo no tenía ocupación de riesgo epidemiológico y la comorbilidad pulmonar se detectó en 97,9 %, sobresaliendo la tuberculosis pulmonar. Se comprobó la no correspondencia entre la terapéutica recibida y el régimen recomendado por la American Thoracic Society, en 100 % de los pacientes. Conclusión: la instauración de un programa ajustado a las normativas de tratamiento internacionalmente aceptadas, constituye en el Hospital Benéfico Jurídico, una necesidad para la atención a este grupo de enfermos.Introduction: the global concern about diseases caused by environmental mycobacteria is growing since their diagnosis and treatment are difficult. Objectives: to determine the behavior of Mycobacterium avium intracellulare complex pulmonary disease, and the therapeutic match with the

  16. Alternaria cerasidanica sp nov., isolated in Denmark from drupes of Prunus avium

    DEFF Research Database (Denmark)

    Roberts, R. G.; Reymond, S. T.; Andersen, Birgitte

    2010-01-01

    fingerprints of A. cerasidanica and 167 strains of Alternaria spp. support the placement of A. cerasidanica within the A. infectoria species-group sensu Simmons and its segregation from other members of this group. A. cerasidanica is currently monotypic and known only from preharvest sweet cherry fruit in......The ex-type strain of Alternaria cerasidanica was isolated in 2001 from an immature, asymptomatic drupe of Prunus avium collected at a commercial cherry orchard near Skaelskor, Denmark. Cultural morphology, sporulation pattern and cluster analyses of combined RAPD, RAMS (microsatellite), and AFLP...

  17. Mycobacterium avium complex-associated peritonitis with CAPD after unrelated bone marrow transplantation.

    Science.gov (United States)

    Miyashita, Emiko; Yoshida, Hisao; Mori, Daisuke; Nakagawa, Natsuki; Miyamura, Takako; Ohta, Hideaki; Seki, Masafumi; Tomono, Kazunori; Hashii, Yoshiko; Ozono, Keiichi

    2014-12-01

    Peritonitis remains an important complication of peritoneal dialysis and is mostly caused by aerobic enteric bacteria. Non-tuberculous mycobacteria (NTM)-associated peritonitis is an unusual but serious infection, requiring special culture techniques to avoid delay in diagnosis. We report the case of an 11-year-old girl with aplastic anemia on ambulatory peritoneal dialysis who had Mycobacterium avium complex-associated peritonitis after allogeneic hematopoietic stem cell transplantation (allo-HSCT). This case emphasizes that we should be constantly cautious about NTM infection in allo-HSCT recipients, especially when standard cultures are negative and the infection is refractory to empirical antibiotic therapy. PMID:25521993

  18. 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.

  19. La cryoconservation des semences dormantes orthodoxes des arbres forestiers : le merisier (Prunus avium L.)

    OpenAIRE

    Chmielarz, Paweł

    2009-01-01

    International audience * The sensitivity of dormant seeds of mazzard cherry (Prunus avium L.) of Polish provenances to extreme desiccation and/or the ultra-low temperature of liquid nitrogen, LN (− 196 °C), was investigated to identify the optimum seed water content (w.c.) at this temperature.* Germination and seedling emergence tests in this study showed that there is no critical seed w.c. for deeply desiccated seeds (stone w.c. 1.6%), although such desiccated seeds were sensitive to the ...

  20. MIKRORAZMNOŽAVANJE DIVLJE TREŠNJE (Prunus avium L.) IZ KLONSKE SJEMENSKE PLANTAŽE

    OpenAIRE

    TANČEVA CRMARIĆ, Olivera; Kajba, Davorin

    2016-01-01

    Velika pozornost pridaje se divljoj trešnji (Prunus avium L.) zbog plemenitog i visokovrijednog drva, ali i zbog važnosti očuvanja njezine genetičke raznolikosti. Oplemenjivanje divlje trešnje uobičajenim metodama uzgoja mogao bi biti vrlo spor proces, otežan time da je u prirodi teško dobiti redovne i dovoljne količine sjemena. Iako osnivanje klonskih sjemenskih plantaža osigurava redovitiji prinos kvalitetnog sjemena, ipak ostaje ovisnost od vremenskih prilika koje utječu na cvjetanje tj. p...

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

    International Nuclear Information System (INIS)

    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

  2. Effect of breed composition, temperament, and ELISA scores for paratuberculosis on phenotypic residual feed intake and growth in an Angus-Brahman multibreed herd.

    Science.gov (United States)

    Breed composition, temperament, and subclinical paratuberculosis in dams are factors that may have an effect on growth and feed efficiency in beef cattle. The objective of this research was to assess the effect of breed group (Angus (A), Brahman (B), Brangus, 3/4 A 1/4 B, 1/2 A ½ B, and 1/4 A 3/4 B)...

  3. Evolution of Compatibility Range in the Rice-Magnaporthe oryzae System: An Uneven Distribution of R Genes Between Rice Subspecies.

    Science.gov (United States)

    Gallet, Romain; Fontaine, Colin; Bonnot, François; Milazzo, Joëlle; Tertois, Christophe; Adreit, Henri; Ravigné, Virginie; Fournier, Elisabeth; Tharreau, Didier

    2016-04-01

    Efficient strategies for limiting the impact of pathogens on crops require a good understanding of the factors underlying the evolution of compatibility range for the pathogens and host plants, i.e., the set of host genotypes that a particular pathogen genotype can infect and the set of pathogen genotypes that can infect a particular host genotype. Until now, little is known about the evolutionary and ecological factors driving compatibility ranges in systems implicating crop plants. We studied the evolution of host and pathogen compatibility ranges for rice blast disease, which is caused by the ascomycete Magnaporthe oryzae. We challenged 61 rice varieties from three rice subspecies with 31 strains of M. oryzae collected worldwide from all major known genetic groups. We determined the compatibility range of each plant variety and pathogen genotype and the severity of each plant-pathogen interaction. Compatibility ranges differed between rice subspecies, with the most resistant subspecies selecting for pathogens with broader compatibility ranges and the least resistant subspecies selecting for pathogens with narrower compatibility ranges. These results are consistent with a nested distribution of R genes between rice subspecies. PMID:26667186

  4. Genetic relationships among some subspecies of the Peregrine Falcon (Falco peregrinus L.), inferred from mitochondrial DNA control-region sequences

    Science.gov (United States)

    White, Clayton M.; Sonsthagen, Sarah A.; Sage, George K.; Anderson, Clifford; Talbot, Sandra L.

    2013-01-01

    The ability to successfully colonize and persist in diverse environments likely requires broad morphological and behavioral plasticity and adaptability, and this may partly explain why the Peregrine Falcon (Falco peregrinus) exhibits a large range of morphological characteristics across their global distribution. Regional and local differences within Peregrine Falcons were sufficiently variable that ∼75 subspecies have been described; many were subsumed, and currently 19 are generally recognized. We used sequence information from the control region of the mitochondrial genome to test for concordance between genetic structure and representatives of 12 current subspecies and from two areas where subspecies distributions overlap. Haplotypes were broadly shared among subspecies, and all geographic locales shared a widely distributed common haplotype (FalconCR2). Haplotypes were distributed in a star-like phylogeny, consistent with rapid expansion of a recently derived species, with observed genetic patterns congruent with incomplete lineage sorting and/or differential rates of evolution on morphology and neutral genetic characters. Hierarchical analyses of molecular variance did not uncover genetic partitioning at the continental level, despite strong population-level structure (FST = 0.228). Similar analyses found weak partitioning, albeit significant, among subspecies (FCT = 0.138). All reconstructions placed the hierofalcons' (Gyrfalcon [F. rusticolus] and Saker Falcon [F. cherrug]) haplotypes in a well-supported clade either basal or unresolved with respect to the Peregrine Falcon. In addition, haplotypes representing Taita Falcon (F. fasciinucha) were placed within the Peregrine Falcon clade.

  5. Identical metabolic rate and thermal conductance in Rock Sandpiper (Calidris ptilocnemis) subspecies with contrasting nonbreeding life histories

    Science.gov (United States)

    Ruthrauff, Daniel R.; Dekinga, Anne; Gill, Robert E., Jr.; Piersma, Theunis

    2013-01-01

    Closely related species or subspecies can exhibit metabolic differences that reflect site-specific environmental conditions. Whether such differences represent fixed traits or flexible adjustments to local conditions, however, is difficult to predict across taxa. The nominate race of Rock Sandpiper (Calidris ptilocnemis) exhibits the most northerly nonbreeding distribution of any shorebird in the North Pacific, being common during winter in cold, dark locations as far north as upper Cook Inlet, Alaska (61°N). By contrast, the tschuktschorum subspecies migrates to sites ranging from about 59°N to more benign locations as far south as ~37°N. These distributional extremes exert contrasting energetic demands, and we measured common metabolic parameters in the two subspecies held under identical laboratory conditions to determine whether differences in these parameters are reflected by their nonbreeding life histories. Basal metabolic rate and thermal conductance did not differ between subspecies, and the subspecies had a similar metabolic response to temperatures below their thermoneutral zone. Relatively low thermal conductance values may, however, reflect intrinsic metabolic adaptations to northerly latitudes. In the absence of differences in basic metabolic parameters, the two subspecies’ nonbreeding distributions will likely be more strongly influenced by adaptations to regional variation in ecological factors such as prey density, prey quality, and foraging habitat.

  6. Genetic structure and subspecies divergence of wild boars Sus scrofa in mainland China based on the microsatellite variation analysis

    Directory of Open Access Journals (Sweden)

    ZHANG Bao-Wei

    2008-12-01

    Full Text Available In order to discuss the subspecies classification of wild boar Sus scrofa in China, We studied their genetic structure using 11 microsatellites, with emphasis on South China. High levels of genetic variationwere detected in South China (SC, North China (NC and Northeast China (NE wild boar populations. This was consistent with the huge population size of the wild boar and its status as a widely-distributed species. Weak genetic divergence was observed in all subspecies involved in this study. Individuals from all subspecies were highly mixed in the phylogeny tree based on the allele sharing distance (DAS, no distinct clade was found according to the defined subspecies. Even considering the two main factors, which reflect the biggest genetic differentiation among the populations, it is impossible to distinguish SC,NC and NE from each other by FCA (Factor Correspondence Analysis. Using the F-statistic test, weak genetic differentiation (Fst = 0.014 was detected between SC and NC population, which indicated a high gene flow between two sides of the Yangtze River. The Yangtze River is an insurmountable geographical hindrance for wild boars. The stronger population differentiations were observed between NE and NC population, and between NE and SC population(Fst = 0.040, 0.042 respectively), this may suggest that the wild boars in NE area could be defined as the subspecies S. s. ussuricus.[Acta Zoologica Sinica 54(5: 753 – 761, 2008].

  7. Morpho- biochemical evaluation of Brassica rapa sub-species for salt tolerance

    Directory of Open Access Journals (Sweden)

    Jan Sohail Ahmad

    2016-01-01

    Full Text Available Salt stress is one of the key abiotic stresses that affect both the qualitative and quantitative characters of many Brassica rapa sub-species by disturbing its normal morphobiochemical processes. Therefore, the present research work was designed to study the effect of different NaCl events (0, 50,100 and 150 mmol on morphological and biochemical characters and to screen salt tolerant genotypes among brown, yellow and toria types of B. rapa sub-species. The plants were grown in test tubes with addition of four level of NaCl (0, 50,100 and 150 mmol. The effect of salinity on shoot and root length, shoot/ root fresh and dry weight, relative water content (RWC, proline and chlorophyll a, b, a+b contents was recorded after 4 weeks of sowing. The genotype 22861 (brown type showed excellent morphological and biochemical performance at all stress levels followed by Toria-Sathi and Toria-A respectively as compared to Check variety TS-1. The genotype 26158 (yellow type gave very poor performance and retard growth. The %RWC values and chlorophyll a, b and a+b contents were decreased several folds with the increase of salt concentration. While, the proline contents was increased with raising of salt stress. The brown and toria types showed maximum tolerance to salt stress at early germination stages as compare to yellows one. The present study will serve as model to develop quick salt tolerant genotypes among different plant sub-species against salt stress.

  8. A molecular marker distinguishes the subspecies Melipona quadrifasciata quadrifasciata and Melipona quadrifasciata anthidioides (Hymenoptera: Apidae, Meliponinae

    Directory of Open Access Journals (Sweden)

    Ana M. Waldschmidt

    2000-09-01

    Full Text Available The stingless bee species Melipona quadrifasciata includes two subspecies, Melipona quadrifasciata anthidioids and Melipona quadrifasciata quadrifasciata. The morphological difference between the two subspecies is the presence of three to five continuous yellow stripes on the terga on the 3rd to 6th segments in workers and males of M. q. quadrifasciata, and two to five interrupted bands in M. q. anthidioides. We identified a DNA marker which is present in M. q. quadrifasciata and absent in M. q. anthidioides. Only one among the M. q. quadrifasciata colonies did not present the marker. It was also absent in bees collected in northern Minas Gerais State (Brazil, despite their morphological resemblance to M. q. quadrifasciata. The marker can be used for studying the genetic structure of the hybridization zone formed by the intercrossing of the two subspecies.A espécie de abelha sem ferrão Melipona quadrifasciata apresenta duas subespécies, Melipona quadrifasciata quadrifasciata Lep. e Melipona quadrifasciata anthidioides Lep. A diferença morfológica entre as duas subespécies é a presença de três a cinco bandas tergais amarelas do 3º ao 6º segmentos em operárias e machos de M. q. quadrifasciata e duas a cinco bandas interrompidas em M. q. anthidioides. Nós identificamos um marcador de DNA que está presente em M. q. quadrifasciata e ausente em M. q. anthidioides. Este marcador está ausente em abelhas coletadas no norte do Estado de Minas Gerais (Brasil, embora esses indivíduos apresentem morfologia similar à de M. q. quadrifasciata. Este marcador poderá ser utilizado em estudos da zona de hibridação entre as subespécies.

  9. Sub-specialization in plastic surgery in Sub-saharan Africa: capacities, gaps and opportunities

    OpenAIRE

    Ibrahim, Abdulrasheed

    2014-01-01

    The skill set of a plastic surgeon, which addresses a broad range of soft tissue conditions that are prevalent in sub-Saharan Africa, remains relevant in the unmet need for surgical care. Recently, there has being a major paradigm shift from discipline-based to disease-based care, resulting in an emerging component of patient-centered care; adequate access to subspecialty care in plastic and reconstructive surgery. Given the need for an evolution in sub-specialization, this article focuses on...

  10. USE OF A NATURAL DYE FROM SERRATIA MARCESCENS SUBSPECIES MARCESCENS IN DYEING OF TEXTILE FABRICS

    Directory of Open Access Journals (Sweden)

    Ravindra Adivarekar

    2013-06-01

    Full Text Available A strain of Serratia marcescens subspecies marcescens capable of producing a novel rose red pigment with a mass of 112 Da has been isolated from Mahim Mangroove soil. Studies regarding the growth conditions of bacteria, partial characterization of the produced pigment and use of this rose red pigment to dye natural fabrics has been studied and described. Dyeing of wool, cotton and silk fabrics with this rose red microbial pigment as natural dye indicated that the colour strength values and the dye uptake were high with satisfactory fastness properties of the dyed fabric.

  11. Transcriptome characterization and polymorphism detection between subspecies of big sagebrush (Artemisia tridentata

    Directory of Open Access Journals (Sweden)

    Cronn Richard C

    2011-07-01

    Full Text Available Abstract Background Big sagebrush (Artemisia tridentata is one of the most widely distributed and ecologically important shrub species in western North America. This species serves as a critical habitat and food resource for many animals and invertebrates. Habitat loss due to a combination of disturbances followed by establishment of invasive plant species is a serious threat to big sagebrush ecosystem sustainability. Lack of genomic data has limited our understanding of the evolutionary history and ecological adaptation in this species. Here, we report on the sequencing of expressed sequence tags (ESTs and detection of single nucleotide polymorphism (SNP and simple sequence repeat (SSR markers in subspecies of big sagebrush. Results cDNA of A. tridentata sspp. tridentata and vaseyana were normalized and sequenced using the 454 GS FLX Titanium pyrosequencing technology. Assembly of the reads resulted in 20,357 contig consensus sequences in ssp. tridentata and 20,250 contigs in ssp. vaseyana. A BLASTx search against the non-redundant (NR protein database using 29,541 consensus sequences obtained from a combined assembly resulted in 21,436 sequences with significant blast alignments (≤ 1e-15. A total of 20,952 SNPs and 119 polymorphic SSRs were detected between the two subspecies. SNPs were validated through various methods including sequence capture. Validation of SNPs in different individuals uncovered a high level of nucleotide variation in EST sequences. EST sequences of a third, tetraploid subspecies (ssp. wyomingensis obtained by Illumina sequencing were mapped to the consensus sequences of the combined 454 EST assembly. Approximately one-third of the SNPs between sspp. tridentata and vaseyana identified in the combined assembly were also polymorphic within the two geographically distant ssp. wyomingensis samples. Conclusion We have produced a large EST dataset for Artemisia tridentata, which contains a large sample of the big sagebrush

  12. SRI-286, a Thiosemicarbazole, in Combination with Mefloquine and Moxifloxacin for Treatment of Murine Mycobacterium avium Complex Disease

    OpenAIRE

    Bermudez, Luiz E.; Kolonoski, Peter; Seitz, Lianne E.; Petrofsky, Mary; Reynolds, Robert; Wu, Martin; Young, Lowell S.

    2004-01-01

    Treatment of Mycobacterium avium disease remains challenging when macrolide resistance develops. We infected C57 beige mice and treated them with mefloquine, SRI-286, and moxifloxacin. SRI-286 (80 mg/kg) was bactericidal in the liver. Mefloquine plus moxifloxacin or mefloquine plus SRI-286 were better than mefloquine alone.

  13. Clinal variation or validation of a subspecies? A case study of the Graptemys nigrinoda complex (Testudines: Emydidae)

    Science.gov (United States)

    Ennen, Joshua R.; Kalis, Marley E.; Patterson, Adam L.; Kreiser, Brian R.; Lovich, Jeffrey E.; Godwin, James; Qualls, Carl P.

    2014-01-01

    Widely distributed species often display intraspecific morphological variation due to the abiotic and biotic gradients experienced across their ranges. Historically, in many vertebrate taxa, such as birds and reptiles, these morphological differences within a species were used to delimit subspecies. Graptemys nigrinoda is an aquatic turtle species endemic to the Mobile Bay Basin. Colour pattern and morphological variability were used to describe a subspecies (G. n. delticola) from the lower reaches of the system, although it and the nominate subspecies also reportedly intergrade over a large portion of the range. Other researchers have suggested that these morphological differences merely reflect clinal variation. Our molecular data (mtDNA) did not support the existence of the subspecies, as the haplotypes were differentiated by only a few base pairs and one haplotype was shared between the putative subspecies. While there were significant morphological and pattern differences among putative specimens of G. n. nigrinoda, G. n. delticola and G. n. nigrinoda × delticola, these differences probably represent clinal variation as they were also related to environmental variables [i.e. cumulative drainage area and drainage (categorical)]. Specimens occupying slow-current, high-turbidity river reaches (e.g. the Tensaw River) exhibited greater relative carapace heights and more dark pigmentation, while specimens occupying fast-current, clearer rivers (e.g. the upper Alabama, Cahaba and Tallapoosa rivers) exhibited lower carapace heights and more yellow pigmentation. Given the absence of clear molecular and morphological differences that are related to drainage characteristics, we suggest that there is not sufficient evidence for the recognition of G. n. delticola as a distinct subspecies.

  14. Chemical Diversity in Volatiles of Helichrysum plicatum DC. Subspecies in Turkey

    Directory of Open Access Journals (Sweden)

    Bintuğ Öztürk

    2014-07-01

    Full Text Available In the present work three subspecies of Helichrysum plicatum DC. (Helichrysum plicatum DC. subsp. plicatum, Helichrysum plicatum DC. subsp. polyphillum (Ledeb P.H.Davis & Kupicha and Helichrysum plicatum DC. subsp. isauricum Parolly were investigated for the essential oil chemical compositions. The volatiles were obtained by conventional hydrodistillation of aerial parts and microdistillation of inflorescences. Subsequent gas chromatography (GC-FID and gas chromatography coupled to mass spectrometry (GC/MS revealed chemical diversity in compositions of the volatiles analyzed. A total of 199 compounds were identified representing 73.9-98.3% of the volatiles compositions. High abundance of fatty acids and their esters (24.9-70.8% was detected in the herb volatiles of H. plicatum subsp. polyphyllum and H. plicatum subsp. isauricum. The inflorescences of Helichrysum subspecies were found to be rich in monoterpenes (15.0-93.1%, fatty acids (0.1-36.3% and sesquiterpenes (1.1-25.5%. The inflorescence volatiles of H. plicatum subsp. isauricum were distinguished by predomination of monoterpene hydrocarbons (93.1% with fenchene (88.3% as the major constituent

  15. Disentangling canid howls across multiple species and subspecies: Structure in a complex communication channel.

    Science.gov (United States)

    Kershenbaum, Arik; Root-Gutteridge, Holly; Habib, Bilal; Koler-Matznick, Janice; Mitchell, Brian; Palacios, Vicente; Waller, Sara

    2016-03-01

    Wolves, coyotes, and other canids are members of a diverse genus of top predators of considerable conservation and management interest. Canid howls are long-range communication signals, used both for territorial defence and group cohesion. Previous studies have shown that howls can encode individual and group identity. However, no comprehensive study has investigated the nature of variation in canid howls across the wide range of species. We analysed a database of over 2000 howls recorded from 13 different canid species and subspecies. We applied a quantitative similarity measure to compare the modulation pattern in howls from different populations, and then applied an unsupervised clustering algorithm to group the howls into natural units of distinct howl types. We found that different species and subspecies showed markedly different use of howl types, indicating that howl modulation is not arbitrary, but can be used to distinguish one population from another. We give an example of the conservation importance of these findings by comparing the howls of the critically endangered red wolves to those of sympatric coyotes Canis latrans, with whom red wolves may hybridise, potentially compromising reintroduced red wolf populations. We believe that quantitative cross-species comparisons such as these can provide important understanding of the nature and use of communication in socially cooperative species, as well as support conservation and management of wolf populations. PMID:26809021

  16. Karyotypic evolution of ribosomal sites in buffalo subspecies and their crossbreed

    Directory of Open Access Journals (Sweden)

    Tiago Marafiga Degrandi

    2014-06-01

    Full Text Available Domestic buffaloes are divided into two group based on cytogenetic characteristics and habitats: the "river buffaloes" with 2n = 50 and the "swamp buffaloes", 2n = 48. Nevertheless, their hybrids are viable, fertile and identified by a 2n = 49. In order to have a better characterization of these different cytotypes of buffaloes, and considering that NOR-bearing chromosomes are involved in the rearrangements responsible for the karyotypic differences, we applied silver staining (Ag-NOR and performed fluorescent in situ hybridization (FISH experiments using 18S rDNA as probe. Metaphases were obtained through blood lymphocyte culture of 21 individuals, including river, swamp and hybrid cytotypes. Ag-NOR staining revealed active NORs on six chromosome pairs (3p, 4p, 6, 21, 23, 24 in the river buffaloes, whereas the swamp buffaloes presented only five NOR-bearing pairs (4p, 6, 20, 22, 23. The F1 crossbreed had 11 chromosomes with active NORs, indicating expression of both parental chromosomes. FISH analysis confirmed the numerical divergence identified with Ag-NOR. This result is explained by the loss of the NOR located on chromosome 4p in the river buffalo, which is involved in the tandem fusion with chromosome 9 in this subspecies. A comparison with the ancestral cattle karyotype suggests that the NOR found on the 3p of the river buffalo may have originated from a duplication of ribosomal genes, resulting in the formation of new NOR sites in this subspecies.

  17. Electrocardiography in two subspecies of manatee (Trichechus manatus latirostris and T. m. manatus).

    Science.gov (United States)

    Siegal-Willott, Jessica; Estrada, Amara; Bonde, Robert; Wong, Arthur; Estrada, Daniel J; Harr, Kendal

    2006-12-01

    Electrocardiographic (ECG) measurements were recorded in two subspecies of awake, apparently healthy, wild manatees (Trichechus manatus latirostris and T. m. manatus) undergoing routine field examinations in Florida and Belize. Six unsedated juveniles (dependent and independent calves) and 6 adults were restrained in ventral recumbency for ECG measurements. Six lead ECGs were recorded for all manatees and the following parameters were determined: heart rate and rhythm; P, QRS, and T wave morphology, amplitude, and duration; and mean electrical axis (MEA). Statistical differences using a t-test for equality of means were determined. No statistical difference was seen based on sex or subspecies of manatees in the above measured criteria. Statistical differences existed in heart rate (P = 0.047), P wave duration (P = 0.019), PR interval (P = 0.025), and MEA (P = 0.021) between adult manatees and calves. Our findings revealed normal sinus rhythms, no detectable arrhythmias, prolonged PR and QT intervals, prolonged P wave duration, and small R wave amplitude as compared with cetacea and other marine mammals. This paper documents the techniques for and baseline recordings of ECGs in juvenile and adult free-living manatees. It also demonstrates that continual assessment of cardiac electrical activity in the awake manatee can be completed and can be used to aid veterinarians and biologists in routine health assessment, during procedures, and in detecting the presence of cardiac disease or dysfunction. PMID:17315428

  18. Electrocardiography in two subspecies of manatee (Trichechus manatus latirostris and Trichechus manatus manatus)

    Science.gov (United States)

    Siegal-Willott, J.; Estrada, A.; Bonde, R.K.; Wong, A.; Estrada, D.J.; Harr, K.

    2006-01-01

    Electrocardiographic (ECG) measurements were recorded in two subspecies of awake, apparently healthy, wild manatees (Trichechus manatus latirostris and T. m. manatus) undergoing routine field examinations in Florida and Belize. Six unsedated juveniles (dependent and independent calves) and 6 adults were restrained in ventral recumbency for ECG measurements. Six lead ECGs were recorded for all manatees and the following parameters were determined: heart rate and rhythm; P, QRS, and T wave morphology, amplitude, and duration; and mean electrical axis (MEA). Statistical differences using a t-test for equality of means were determined. No statistical difference was seen based on sex or subspecies of manatees in the above measured criteria. Statistical differences existed in heart rate (P = 0.047), P wave duration (P = 0.019), PR interval (P = 0.025), and MEA (P = 0.021) between adult manatees and calves. Our findings revealed normal sinus rhythms, no detectable arrhythmias, prolonged PR and QT intervals, prolonged P wave duration, and small R wave amplitude as compared with cetacea and other marine mammals. This paper documents the techniques for and baseline recordings of ECGs in juvenile and adult free-living manatees. It also demonstrates that continual assessment of cardiac electrical activity in the awake manatee can be completed and can be used to aid veterinarians and biologists in routine health assessment, during procedures, and in detecting the presence of cardiac disease or dysfunction.

  19. Direct identification of Photobacterium damselae subspecies piscicida by PCR-RFLP analysis.

    Science.gov (United States)

    Zappulli, V; Patarnello, T; Patarnello, P; Frassineti, F; Franch, R; Manfrin, A; Castagnaro, M; Bargelloni, L

    2005-06-14

    Fish pasteurellosis is an infectious disease that affects several teleost species living in temperate marine waters. The pathogen responsible, Photobacterium damselae subspecies piscicida, shows high genetic similarity with P. damselae subsp. damselae, making subspecies discrimination extremely laborious. Here we report for the first time a PCR-RFLP method for the identification of P. damselae subsp. piscicida without prior isolation in pure culture. Genomic sequence information was obtained through cloning and sequencing of RAPD products. Two P. damselae-specific primer pairs were developed and tested on 17 strains of P. damselae subsp. piscicida, 10 strains of P. damselae subsp. damselae, and 6 closely related control species. High sensitivity was achieved in PCR amplification on serially diluted samples (analysis of PCR products showed a unique digestion profile for all P. damselae subsp. piscicida strains. The same PCR-RFLP method was implemented on total DNA samples extracted from experimentally infected sea bream and sea bass. Positive results were obtained on fish with clear signs of the disease as well as on challenged, but asymptomatic, fish. The method presented here might provide a useful tool for both prevention and rapid diagnosis of fish pasteurellosis. PMID:16042043

  20. Genome Sequences of Four Strains of Mycobacterium avium subsp. hominissuis, Isolated from Swine and Humans, Differing in Virulence in a Murine Intranasal Infection Model

    Science.gov (United States)

    Bruffaerts, N.; Vluggen, C.; Duytschaever, L.; Mathys, V.; Saegerman, C.; Chapeira, O.

    2016-01-01

    This paper announces the genome sequences of four strains of Mycobacterium avium subsp. hominissuis, isolated from cases of lymphadenopathy in swine and humans, differing in virulence in a murine intranasal infection model. PMID:27313293

  1. Single nucleotide polymorphism discovery in cutthroat trout subspecies using genome reduction, barcoding, and 454 pyro-sequencing

    Directory of Open Access Journals (Sweden)

    Houston Derek D

    2012-12-01

    Full Text Available Abstract Background Salmonids are popular sport fishes, and as such have been subjected to widespread stocking throughout western North America. Historically, stocking was done with little regard for genetic variation among populations and has resulted in genetic mixing among species and subspecies in many areas, thus putting the genetic integrity of native salmonid populations at risk and creating a need to assess the genetic constitution of native salmonid populations. Cutthroat trout is a salmonid species with pronounced geographic structure (there are 10 extant subspecies and a recent history of hybridization with introduced rainbow trout in many populations. Genetic admixture has also occurred among cutthroat trout subspecies in areas where introductions have brought two or more subspecies into contact. Consequently, management agencies have increased their efforts to evaluate the genetic composition of cutthroat trout populations to identify populations that remain uncompromised and manage them accordingly, but additional genetic markers are needed to do so effectively. Here we used genome reduction, MID-barcoding, and 454-pyrosequencing to discover single nucleotide polymorphisms that differentiate cutthroat trout subspecies and can be used as a rapid, cost-effective method to characterize the genetic composition of cutthroat trout populations. Results Thirty cutthroat and six rainbow trout individuals were subjected to genome reduction and next-generation sequencing. A total of 1,499,670 reads averaging 379 base pairs in length were generated by 454-pyrosequencing, resulting in 569,060,077 total base pairs sequenced. A total of 43,558 putative SNPs were identified, and of those, 125 SNP primers were developed that successfully amplified 96 cutthroat trout and rainbow trout individuals. These SNP loci were able to differentiate most cutthroat trout subspecies using distance methods and Structure analyses. Conclusions Genomic and

  2. Correlation of antigen-specific IFN-γ responses of fresh blood samples from Mycobacterium avium subsp. paratuberculosis infected heifers with responses of day-old samples co-cultured with IL-12 or anti-IL-10 antibodies

    DEFF Research Database (Denmark)

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

    2012-01-01

    overnight with specific MAP antigens followed by quantification of IFN-γ by ELISA. It is recommended that the time interval from sampling to culture does not exceed eight hours but addition of the co-stimulating cytokine interleukin 12 (IL-12) or anti-IL-10 antibodies to culture have been demonstrated to...... enhance IFN-γ responses of cultures stimulated with Johnin purified protein derivative (PPDj). Here we examined the correlation of IFN-γ production in response to PPDj and 15 recombinant antigens in day-old blood samples from heifers 10–21 months of age from a MAP infected herd with addition of either...... recombinant bovine IL-12 or anti-bovine IL-10 antibody with IFN-γ production in sample day samples. IFN-γ responses of sample day samples showed high correlation with responses to some antigens in day-old samples with addition of IL-12 or anti-IL-10 antibodies to cultures, indicating that day-old protocols...

  3. Identification of genetic markers to distinguish the virulent and avirulent subspecies of Pantoea stewartii by comparative proteomics and genetic analysis.

    Science.gov (United States)

    Wu, Qiong; Jiang, Zide; Liao, Jinliang; Chen, Zhinan; Li, Huaping; Mei, Mantong; Zhang, Lian-Hui

    2007-02-01

    Pantoea stewartii subsp. stewartii (Pnss), the causal agent of Stewart's bacterial wilt and leaf blight of maize and sweet corn, is one of the quarantine pathogens in many countries and regions. In contrast, P. stewartii subsp. indologenes (Pnsi), the closely related subspecies of Pnss, is avirulent on these plants. In this study, the protein expression profiles of these two subspecies were compared using two-dimensional gel electrophoresis analysis. Twenty-one unique protein spots consistently detected in Pnss but not in Pnsi were analyzed by mass spectrometry. Some of these Pnss-specific proteins are known to be essential for virulence and survival in host, such as FoxR and HrcJ, which are the key components of iron uptake and Type III secretion systems, respectively. For further genetic analysis, six Pnss-specific proteins were characterized by peptide sequencing. Southern and Northern blot analyses revealed that the differences in protein expression profiles of the two subspecies were either due to the discrepancy at genome level or because of the variations in transcriptional expression. The results provide novel genetic markers to distinguish the two closely related subspecies and may also serve as useful clues for investigation of the genetic basis accounting for their sharp difference in virulence. PMID:17086414

  4. Similar local, but different systemic, metabolomics responses of closely related pine subspecies to folivory by caterpillars of the processionary moth

    Energy Technology Data Exchange (ETDEWEB)

    Rivas Ubach, Albert; Sardans, J.; Hodar, Jose A.; Garcia-Porta, Joan; Guenther, Alex B.; Oravec, Michal; Urban, Otmar; Penuelas, J.

    2016-05-16

    Plants respond locally and systemically to herbivore attack. Most of the research conducted on plant-herbivore relationships at elemental and molecular levels have focused on nutrients or/and certain molecular compounds or specific families of defensive metabolites showing that herbivores tend to select plant individuals or species with higher nutrient concentrations and to avoid those with higher levels of phenolics and terpenes. Unfortunately, the defensive role of phenolics in conifers is still unclear. We performed stoichiometric and metabolomics, local and systemic, analyses in two subspecies of Pinus sylvestris under the herbivorous attack by the caterpillars of the pine processionary moth, an important pest in the Mediterranean Basin. Herbivorous attack was not associated with any of the elements analyzed. Both pine subspecies responded locally to folivory mainly by increasing the concentrations of various terpenes and phenolics. Systemic responses differed between subspecies and most of the metabolites presented intermediate concentrations between those of the affected parts and unattacked trees. Contrary as usually thought, foliar nutrient concentrations did not show to be a main factor of an alleged plant selection by adult female processionary moths for oviposition. Local increases in phenolics were more associated with antioxidant function for protection against oxidative damage produced by folivory. On the other hand, terpenes were directly related to defense against herbivores. Herbivory attack produced a general systemic shift in pines, including both primary and secondary metabolisms, that was, however, less intense and chemically different from the local responses. Subspecies responded similarly locally but differently to folivory at systemic level.

  5. Use of intron-exonic marker in assessment of genetic diversity of two subspecies of Thymus daenensis

    Directory of Open Access Journals (Sweden)

    Ahmad Ismaili

    2013-11-01

    Full Text Available Study of genetic diversity in medicinal plant is very important for improvement and evolutionary variations. In this study, assessment of genetic diversity in two subspecies of Thymus daenensis was evaluated, using intron-exonic markers. Thirty primers produced 633 polymorphic bands (98% polymorphism. The highest polymorphic information content (PIC included ISJ5 and ISJ9 primers and the lowest PIC also included IT15-32 primer. The highest marker index (MI produced by IT10-6 primer. Results of Analysis of Molecular Variance (AMOVA showed that intra-sub specific variation was more than inter-sub specific variation. Dendrogram obtained from Cluster analysis, using NTSYS-pc software and UPGMA method based on Dice's similarity matrix, divided accessions into 4 groups. The maximum range of genetic similarity was observed between two accessions of sub-species daenensis. Two accessions of Fars and Semnan formed a separate group. Results showed that clustering based on molecular data and principal coordinate analysis had a medium alignment. Grouping based on cluster analysis also could separate two subspecies of Thymus daenensis. Results obtained from this study showed that intron-exonic markers had an effective potential in assessment of genetic relationships between the two sub-species of daenensis.

  6. 76 FR 61531 - Endangered and Threatened Wildlife and Plants; 90-Day Finding on a Petition To List 10 Subspecies...

    Science.gov (United States)

    2011-10-04

    ... present information that indicates the degree, if any, that invasive plants are spreading in the... silverspot. carsonensis. In the February 28, 1996, Candidate Notice of Review (CNOR) (61 FR 7595), we adopted... and Threatened Wildlife and Plants; 90-Day Finding on a Petition To List 10 Subspecies of Great...

  7. Distributions of the subspecies of Lesser Black-backed Gulls Larus fuscus in sub-Saharan Africa

    OpenAIRE

    Kylin, Hendrik; Bouwman, Hendrik; Louette, Michel

    2011-01-01

    The wintering area of the nominate subspecies of Lesser Black-backed Gull Larus fuscus fuscus is from Ethiopia across Uganda and the Congo basin to the Atlantic, while L. f. intermedius and L. f. graellsii winter in westernmost Africa. http:/dx.doi.org/10.1080/00063657.2011.555517 The Journal of the British Trust for Ornithology (BTO)

  8. Similar local, but different systemic, metabolomic responses of closely related pine subspecies to folivory by caterpillars of the processionary moth.

    Science.gov (United States)

    Rivas-Ubach, A; Sardans, J; Hódar, J A; Garcia-Porta, J; Guenther, A; Oravec, M; Urban, O; Peñuelas, J

    2016-05-01

    Plants respond locally and systemically to herbivore attack. Most of the research conducted on plant-herbivore relationships at element and molecular levels have focused on the elemental composition or/and certain molecular compounds or specific families of defence metabolites showing that herbivores tend to select plant individuals or species with higher nutrient concentrations and avoid those with higher levels of defence compounds. We performed stoichiometric and metabolomics, both local and systemic, analyses in two subspecies of Pinus sylvestris under attack from caterpillars of the pine processionary moth, an important pest in the Mediterranean Basin. Both pine subspecies responded locally to folivory mainly by increasing relative concentrations of terpenes and some phenolics. Systemic responses differed between pine subspecies, and most of the metabolites presented intermediate concentrations between those of the affected parts and unattacked trees. Our results support the hypothesis that foliar nutrient concentrations are not a key factor for plant selection by adult female processionary moths for oviposition, since folivory was not associated with any of the elements analysed. Phenolic compounds generally did not increase in the attacked trees, questioning the suggestion of induction of phenolics following folivory attack and the anti-feeding properties of phenolics. Herbivory attack produced a general systemic shift in pines, in both primary and secondary metabolism, which was less intense and chemically different from the local responses. Local pine responses were similar between pine subspecies, while systemic responses were more distant. PMID:26642818

  9. Analysis of wild sweet cherry (Prunus avium L. germplasm diversity in south-east Serbia

    Directory of Open Access Journals (Sweden)

    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.

  10. Opportunistic Pathogens Mycobacterium Avium Complex (MAC) and Legionella spp. Colonise Model Shower

    Science.gov (United States)

    Whiley, Harriet; Giglio, Steven; Bentham, Richard

    2015-01-01

    Legionella spp. and Mycobacterium avium complex (MAC) are opportunistic pathogens of public health concern. Hot water systems, including showers, have been identified as a potential source of infection. This paper describes the colonization of Legionella and MAC on the flexible tubing within a model potable shower system, utilizing thermostatic mixing and a flexible shower head. A MAC qPCR method of enumeration was also developed. MAC and Legionella spp. were detected within the biofilm at maximum concentrations of 7.0 × 104 and 2.0 × 103 copies/cm2 PVC tubing respectively. No significant changes were observed between sample of the flexible shower tubing that dried between uses and those that remained filled with water. This suggested the “unhooking” showerheads and allowing them to dry is not an effective method to reduce the risk of Legionella or MAC colonisation. PMID:26213977

  11. Utilization of a ts-sacB selection system for the generation of a Mycobacterium avium serovar-8 specific glycopeptidolipid allelic exchange mutant

    Directory of Open Access Journals (Sweden)

    Belisle John T

    2004-09-01

    Full Text Available Abstract Background Mycobacterium avium are ubiquitous environmental organisms and a cause of disseminated infection in patients with end-stage AIDS. The glycopeptidolipids (GPL of M. avium are proposed to participate in the pathogenesis of this organism, however, establishment of a clear role for GPL in disease production has been limited by the inability to genetically manipulate M. avium. Methods To be able to study the role of the GPL in M. avium pathogenesis, a ts-sacB selection system, not previously used in M. avium, was employed as a means to achieve homologous recombination for the rhamnosyltransferase (rtfA gene of a pathogenic serovar 8 strain of M. avium to prevent addition of serovar-specific sugars to rhamnose of the fatty acyl-peptide backbone of GPL. The genotype of the resultant rtfA mutant was confirmed by polymerase chain reaction and southern hybridization. Disruption in the proximal sugar of the haptenic oligosaccharide resulted in the loss of serovar specific GPL with no change in the pattern of non-serovar specific GPL moieties as shown by thin layer chromatography and gas chromatography/mass spectrometry. Complementation of wild type (wt rtfA in trans through an integrative plasmid restored serovar-8 specific GPL expression identical to wt serovar 8 parent strain. Results In this study, we affirm our results that rtfA encodes an enzyme responsible for the transfer of Rha to 6d-Tal and provide evidence of a second allelic exchange mutagenesis system suitable for M. avium. Conclusion We report the second allelic exchange system for M. avium utilizing ts-sacB as double-negative and xylE as positive counter-selection markers, respectively. This system of allelic exchange would be especially useful for M. avium strains that demonstrate significant isoniazid (INH resistance despite transformation with katG. Through the construction of mutants in GPL or other mycobacterial components, their roles in M. avium pathogenesis

  12. Utilization of a ts-sacB selection system for the generation of a Mycobacterium avium serovar-8 specific glycopeptidolipid allelic exchange mutant

    Science.gov (United States)

    Irani, Vida R; Lee, Sun-Hwa; Eckstein, Torsten M; Inamine, Julia M; Belisle, John T; Maslow, Joel N

    2004-01-01

    Background Mycobacterium avium are ubiquitous environmental organisms and a cause of disseminated infection in patients with end-stage AIDS. The glycopeptidolipids (GPL) of M. avium are proposed to participate in the pathogenesis of this organism, however, establishment of a clear role for GPL in disease production has been limited by the inability to genetically manipulate M. avium. Methods To be able to study the role of the GPL in M. avium pathogenesis, a ts-sacB selection system, not previously used in M. avium, was employed as a means to achieve homologous recombination for the rhamnosyltransferase (rtfA) gene of a pathogenic serovar 8 strain of M. avium to prevent addition of serovar-specific sugars to rhamnose of the fatty acyl-peptide backbone of GPL. The genotype of the resultant rtfA mutant was confirmed by polymerase chain reaction and southern hybridization. Disruption in the proximal sugar of the haptenic oligosaccharide resulted in the loss of serovar specific GPL with no change in the pattern of non-serovar specific GPL moieties as shown by thin layer chromatography and gas chromatography/mass spectrometry. Complementation of wild type (wt) rtfA in trans through an integrative plasmid restored serovar-8 specific GPL expression identical to wt serovar 8 parent strain. Results In this study, we affirm our results that rtfA encodes an enzyme responsible for the transfer of Rha to 6d-Tal and provide evidence of a second allelic exchange mutagenesis system suitable for M. avium. Conclusion We report the second allelic exchange system for M. avium utilizing ts-sacB as double-negative and xylE as positive counter-selection markers, respectively. This system of allelic exchange would be especially useful for M. avium strains that demonstrate significant isoniazid (INH) resistance despite transformation with katG. Through the construction of mutants in GPL or other mycobacterial components, their roles in M. avium pathogenesis, biosynthesis, or drug

  13. Transmission of Streptococcus equi subspecies zooepidemicus infection from horses to humans.

    Science.gov (United States)

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

    2013-07-01

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

  14. Inference of purifying and positive selection in three subspecies of chimpanzees (Pan troglodytes) from exome sequencing

    DEFF Research Database (Denmark)

    Bataillon, Thomas; Duan, Jinjie; Hvilsom, Christina;

    2015-01-01

    of recent gene flow from Western into Eastern chimpanzees. The striking contrast in X-linked vs. autosomal polymorphism and divergence previously reported in Central chimpanzees is also found in Eastern and Western chimpanzees. We show that the direction of selection (DoS) statistic exhibits a strong......We study genome-wide nucleotide diversity in three subspecies of extant chimpanzees using exome capture. After strict filtering, SNVs and indels were called and genotyped for >50% of exons at a mean coverage of 35x per individual. Central chimpanzees (P. t. troglodytes) are the most polymorphic...... (nucleotide diversity, θw= 0.0023 per site) followed by Eastern (P. t. schweinfurthii) chimpanzees (θw = 0.0016) and Western (P. t. verus) chimpanzees (θw = 0.0008). A demographic scenario of divergence without gene flow fits the patterns of autosomal synonymous nucleotide diversity well except for a signal...

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

    Directory of Open Access Journals (Sweden)

    Sasha J Rose

    Full Text Available 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.

  16. 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

  17. 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.

  18. Lipoarabinomanano (LAM de Mycobacterium spp: Respuesta inmune inducida en terneros

    Directory of Open Access Journals (Sweden)

    A. Jolly

    2006-12-01

    Full Text Available La paratuberculosis es una enfermedad que afecta al ganado vacuno cuyo agente etiológico es el Mycobacterium avium subespecie paratuberculosis. El LAM es el principal componente antigénico de superficie de las micobacterias, y se lo considera de relevancia en la patogenia de las enfermedades que éstas causan. Un extracto enriquecido en LAM fue obtenido a partir de un cultivo de Mycobacterium spp. y empleado para inocular terneros. Se evaluó en ellos la respuesta inmune humoral y celular inducida por la vacunación. Los resultados de este estudio demuestran que el extracto enriquecido en LAM resultó ser inmunogénico en todos los animales inoculados, obteniéndose títulos considerables de anticuerpos específicos, sin generar falsos positivos a la prueba de intradermorreacción con el derivado proteico purificado utilizado para el diagnóstico de la tuberculosis bovina. Estos hallazgos justifican continuar el trabajo en esta línea intentando finalmente establecer si el LAM es un candidato potencial para la elaboración de una vacuna a subunidades contra la paratuberculosis bovina.Paratuberculosis is a chronic enteric disease affecting cattle. The causative agent is Mycobacterium avium subspecies paratuberculosis. LAM is the main antigenic component of mycobacterial surface, and it is considered a key virulence factor involved in its pathogenicity. A LAM-enriched extract obtained from a culture of Mycobacterium spp. was prepared with incomplete Freund's adjuvant for calves inoculation. Specific antibodies response and delayed-type hypersensitivity to intradermal injection of purified protein derivative antigen (PPD from Mycobacterium bovis were then evaluated in inoculated animals. Our results demonstrate that anti-LAM antibodies can be successfully obtained in calves immunized with LAM-enriched extract, without generating cross-reaction with PPD of M. bovis. This work could represent the initial step in order to determine the relevance of

  19. Seasonal and geographic variation of southern blue whale subspecies in the Indian Ocean.

    Directory of Open Access Journals (Sweden)

    Flore Samaran

    Full Text Available Understanding the seasonal movements and distribution patterns of migratory species over ocean basin scales is vital for appropriate conservation and management measures. However, assessing populations over remote regions is challenging, particularly if they are rare. Blue whales (Balaenoptera musculus spp are an endangered species found in the Southern and Indian Oceans. Here two recognized subspecies of blue whales and, based on passive acoustic monitoring, four "acoustic populations" occur. Three of these are pygmy blue whale (B.m. brevicauda populations while the fourth is the Antarctic blue whale (B.m. intermedia. Past whaling catches have dramatically reduced their numbers but recent acoustic recordings show that these oceans are still important habitat for blue whales. Presently little is known about the seasonal movements and degree of overlap of these four populations, particularly in the central Indian Ocean. We examined the geographic and seasonal occurrence of different blue whale acoustic populations using one year of passive acoustic recording from three sites located at different latitudes in the Indian Ocean. The vocalizations of the different blue whale subspecies and acoustic populations were recorded seasonally in different regions. For some call types and locations, there was spatial and temporal overlap, particularly between Antarctic and different pygmy blue whale acoustic populations. Except on the southernmost hydrophone, all three pygmy blue whale acoustic populations were found at different sites or during different seasons, which further suggests that these populations are generally geographically distinct. This unusual blue whale diversity in sub-Antarctic and sub-tropical waters indicates the importance of the area for blue whales in these former whaling grounds.

  20. Seasonal and geographic variation of southern blue whale subspecies in the Indian Ocean.

    Science.gov (United States)

    Samaran, Flore; Stafford, Kathleen M; Branch, Trevor A; Gedamke, Jason; Royer, Jean-Yves; Dziak, Robert P; Guinet, Christophe

    2013-01-01

    Understanding the seasonal movements and distribution patterns of migratory species over ocean basin scales is vital for appropriate conservation and management measures. However, assessing populations over remote regions is challenging, particularly if they are rare. Blue whales (Balaenoptera musculus spp) are an endangered species found in the Southern and Indian Oceans. Here two recognized subspecies of blue whales and, based on passive acoustic monitoring, four "acoustic populations" occur. Three of these are pygmy blue whale (B.m. brevicauda) populations while the fourth is the Antarctic blue whale (B.m. intermedia). Past whaling catches have dramatically reduced their numbers but recent acoustic recordings show that these oceans are still important habitat for blue whales. Presently little is known about the seasonal movements and degree of overlap of these four populations, particularly in the central Indian Ocean. We examined the geographic and seasonal occurrence of different blue whale acoustic populations using one year of passive acoustic recording from three sites located at different latitudes in the Indian Ocean. The vocalizations of the different blue whale subspecies and acoustic populations were recorded seasonally in different regions. For some call types and locations, there was spatial and temporal overlap, particularly between Antarctic and different pygmy blue whale acoustic populations. Except on the southernmost hydrophone, all three pygmy blue whale acoustic populations were found at different sites or during different seasons, which further suggests that these populations are generally geographically distinct. This unusual blue whale diversity in sub-Antarctic and sub-tropical waters indicates the importance of the area for blue whales in these former whaling grounds. PMID:23967221

  1. LTR retrotransposons reveal recent extensive inter-subspecies nonreciprocal recombination in Asian cultivated rice

    Directory of Open Access Journals (Sweden)

    Xu Zhao

    2008-11-01

    Full Text Available Abstract Background Long Terminal Repeats retrotransposons (LTR elements are ubiquitous Eukaryotic transposable elements (TEs. They are considered to be one of the major forces underlying plant genome evolution. Because of relatively high evolutionary speed, active transposition of LTR elements in the host genomes provides rich information on their short-term history. As more and more genomes, especially those of closely related organisms, have been sequenced, it is possible to perform global comparative study of their LTR retrotransposons to reveal events in the history. Results The present research is designed to investigate important evolutionary events in the origin of Asian cultivated rice through the comparison of LTR elements. We have developed LTR_INSERT, a new method for LTR elements discovery in two closely related genomes. Our method has a distinctive feature that it is capable of judging whether an insertion occurs prior or posterior to the divergence of genomes. LTR_INSERT identifies 993 full-length LTR elements, annotates 15916 copies related with them, and discovers at least 16 novel LTR families in the whole-genome comparative map of two cultivated rice subspecies. From the full-length LTR elements, we estimate that a significant proportion of the rice genome has experienced inter-subspecies nonreciprocal recombination (ISNR in as recent as 53,000 years. Large-scale samplings further support that more than 15% of the rice genome has been involved in such recombination. In addition, LTR elements confirm that the genome of O. sativa ssp. indica and that of japonica diverged about 600,000 years ago. Conclusion A new LTR retrotransposon identification method integrating both comparative genomics and ab initio algorithm is introduced and applied to Asian cultivated rice genomes. At whole-genome level, this work confirms that recent ISNR is an important factor that molds modern cultivated rice genome.

  2. Foodborne outbreak of Salmonella subspecies IV infections associated with contamination from bearded dragons.

    Science.gov (United States)

    Lowther, S A; Medus, C; Scheftel, J; Leano, F; Jawahir, S; Smith, K

    2011-12-01

    Approximately 1.4 million Salmonella infections and 400 deaths occur annually in the United States. Approximately 6% of human Salmonella cases are thought to be associated with reptiles; Salmonella enterica subspecies IV is primarily reptile-associated. During 1-4 December, 2009, three isolates of Salmonella IV 6,7:z4,z24:- with indistinguishable pulsed-field gel electrophoresis (PFGE) patterns were identified through Minnesota Department of Health laboratory-based surveillance. None of the three patients associated with the isolates reported reptile contact; however, all had attended the same potluck dinner. Dinner attendees were asked questions regarding illness history, foods they prepared for and consumed at the event, and pet ownership. Cases were defined as illness in a person who had eaten potluck food and subsequently experienced fever and diarrhoea (three or more loose stools in 24 h) or laboratory-confirmed infection with Salmonella IV matching the outbreak PFGE subtype. Nineteen days after the event, environmental samples were collected from a food preparer's house where two pet bearded dragons were kept. Sixty-six of 73 potluck food consumers were interviewed; 19 cases were identified; 18 persons reported illness but did not meet the case definition. Median incubation period was 19 h (range: 3-26 h). Median duration of illness was 5 days (range: 1-11 days). Consumption of gravy, prepared by the bearded dragons' asymptomatic owner, was associated with illness (16/32 exposed versus 1/12 unexposed; risk ratio: 6.0; exact P = 0.02). Salmonella Labadi was recovered from 10 samples, including from one bearded dragon, the bathroom door knob and sink drain, and the kitchen sink drain. The outbreak PFGE subtype of Salmonella subspecies IV was isolated from vacuum-cleaner bag contents. This foodborne outbreak probably resulted from environmental contamination from bearded dragons. Reptiles pose a community threat when food for public consumption is prepared in

  3. Mycobacterium avium complex (MAC): an unusual potential pathogen in cerebrospinal fluid of AIDS patients Complexo Mycobacterium avium: um patógeno potencial pouco comum no líquido céfalo -raquidiano de pacientes com AIDS

    OpenAIRE

    David Jamil Hadad; Tereza Cristina Petry; Anaenza Freie Maresca; Lucilaine Ferrazoli; Maria Conceição Martins; Maria Cecilia de Almeida Palhares; Walkyria Pereira Pinto; Adauto Castelo Filho; Moises Palaci

    1995-01-01

    Mycobacterium avium complex (MAC) is frequently isolated from patients with late complications of Acquired Immunodeficiency Syndrome (AIDS), especially in North America and Europe. However, its isolation from the central nervous system (CNS) has been seldom reported in these countries. MAC infections in AIDS patients in African and Latin American countries are believed to be uncommon. We report the isolation of MAC from cerebrospinal fluid (CSF) of 11 AIDS patients out of 1723 (0.63%) seen at...

  4. Cloning and sequencing of a gene encoding a 21-kilodalton outer membrane protein from Bordetella avium and expression of the gene in Salmonella typhimurium.

    Science.gov (United States)

    Gentry-Weeks, C R; Hultsch, A L; Kelly, S M; Keith, J M; Curtiss, R

    1992-12-01

    Three gene libraries of Bordetella avium 197 DNA were prepared in Escherichia coli LE392 by using the cosmid vectors pCP13 and pYA2329, a derivative of pCP13 specifying spectinomycin resistance. The cosmid libraries were screened with convalescent-phase anti-B. avium turkey sera and polyclonal rabbit antisera against B. avium 197 outer membrane proteins. One E. coli recombinant clone produced a 56-kDa protein which reacted with convalescent-phase serum from a turkey infected with B. avium 197. In addition, five E. coli recombinant clones were identified which produced B. avium outer membrane proteins with molecular masses of 21, 38, 40, 43, and 48 kDa. At least one of these E. coli clones, which encoded the 21-kDa protein, reacted with both convalescent-phase turkey sera and antibody against B. avium 197 outer membrane proteins. The gene for the 21-kDa outer membrane protein was localized by Tn5seq1 mutagenesis, and the nucleotide sequence was determined by dideoxy sequencing. DNA sequence analysis of the 21-kDa protein revealed an open reading frame of 582 bases that resulted in a predicted protein of 194 amino acids. Comparison of the predicted amino acid sequence of the gene encoding the 21-kDa outer membrane protein with protein sequences in the National Biomedical Research Foundation protein sequence data base indicated significant homology to the OmpA proteins of Shigella dysenteriae, Enterobacter aerogenes, E. coli, and Salmonella typhimurium and to Neisseria gonorrhoeae outer membrane protein III, Haemophilus influenzae protein P6, and Pseudomonas aeruginosa porin protein F. The gene (ompA) encoding the B. avium 21-kDa protein hybridized with 4.1-kb DNA fragments from EcoRI-digested, chromosomal DNA of Bordetella pertussis and Bordetella bronchiseptica and with 6.0- and 3.2-kb DNA fragments from EcoRI-digested, chromosomal DNA of B. avium and B. avium-like DNA, respectively. A 6.75-kb DNA fragment encoding the B. avium 21-kDa protein was subcloned into the

  5. Subspecies identification of Chimpanzees Pan troglodytes (Primates: Hominidae from the National Zoo of the Metropolitan Park of Santiago, Chile, using mitochondrial DNA sequences

    Directory of Open Access Journals (Sweden)

    J.A. Vega

    2014-05-01

    Full Text Available Natural populations of Chimpanzees Pan troglodytes are declining because of hunting and illegal live animal trafficking. Four subspecies of Chimpanzee have been reported: Pan troglodytes troglodytes, P.t. schweinfurthii, P.t. verus and P.t. ellioti, which have remained geographically separated by natural barriers such as the rivers Niger, Sanaga and Ubangi in central Africa. Sequence analysis of mitochondrial DNA (mtDNA has been used for the determination of these subspecies, which indirectly can also suggest their geographic origin. It was decided to identify the subspecies and the geographic origin of three captive chimpanzees of the National Zoo of the Metropolitan Park of Santiago (Chile, by analyzing their mitochondrial DNA. DNA was extracted from the saliva of three adult chimpanzees (two males and one female. After the analysis of sequences of the mitochondrial hypervariable region (HVI, a phylogenetic tree was constructed using mitochondrial sequences of known Pan troglodytes subspecies. Molecular phylogeny analysis revealed that the chimpanzees are likely to belong to three different subspecies: P.t. schweinfurthii, P.t. verus and P.t. troglodytes. Identification of subspecies of the three chimpanzees of the National Zoo of the Metropolitan Park of Santiago (Chile was possible due to mtDNA analysis. Future identification of chimpanzees will allow the development of a studbook for the chimpanzee subspecies in other Latin American zoos.

  6. Paratuberculosis vaccination causes only limited cross-reactivity in the skin test for diagnosis of bovine tuberculosis.

    Directory of Open Access Journals (Sweden)

    Joseba M Garrido

    Full Text Available Although there is a wide consensus on the efficacy of paratuberculosis vaccination to limit economic losses, its use has been restricted because of its interference in the diagnosis of tuberculosis. Data from a vaccine clinical trial in the Basque Country (Spain has been evaluated in relationship with bovine tuberculosis intradermal test results. The trial included two herds applying a Test and Culling strategy and five applying an inactivated vaccine. The vaccine was applied to animals of all ages present in each vaccinated herd when joining the trial, and then to all the replacers within their first three months of life. Yearly testing done with the comparative intradermal test (CIT was applied to all animals older than 6 weeks. Between 2005 and 2011, the study generated 2,033 records from Vaccinated Herds (VH and 2,252 from Test and Cull herds (TC. Pre-vaccination positive results rate was 2.40% among the 7 herds in the single bovine intradermal tuberculin test (BSIT. Two years later it rose to 20.42% in the VH and remained below at 0.75% in the TC. Applying the CIT reduced these rates to only 0.58% in the VH and to 0.25% in the TC ons. Regarding time since each animal joined the program, the proportion of positives to BSIT was variable and, in some cases, significantly different between time points. With regard to the age of vaccination, no significant differences were found between vaccination within the first year of life and afterwards. Vaccinated animals showed seventeen times more reactions than the non-vaccinated in the BSIT, but only four times more in the CIT. In conclusion, comparative intradermal test can be a useful tool to differentiate paratuberculosis vaccine cross-reactions from specific bovine tuberculosis reactions according to the European and Spanish legislation.

  7. Association between cow reproduction and calf growth traits and ELISA scores for paratuberculosis in a multibreed herd of beef cattle.

    Science.gov (United States)

    Elzo, M A; Rae, D O; Lanhart, S E; Hembry, F G; Wasdin, J G; Driver, J D

    2009-08-01

    The objective of this research was to assess the association between 4 cow reproductive and weight traits, and 2 preweaning calf traits and ELISA scores for paratuberculosis (0 = negative, 1 = suspect, 2 = weak-positive, and 3 = positive) in a multibreed herd of cows ranging from 100% Angus (A) to 100% Brahman (B). Cow data were 624 gestation lengths (GL), 358 records of time open (TO), 605 calving intervals (CI), and 1240 weight changes from November to weaning in September (WC) from 502 purebred and crossbred cows. Calf data consisted of 956 birth weights (BWT), and 923 weaning weights adjusted to 205 d of age (WW205) from 956 purebred and crossbred calves. Traits were analyzed individually using multibreed mixed models that assumed homogeneity of variances across breed groups. Covariances among random effects were assumed to be zero. Fixed effects were year, age of cow, sex of calf, year x age of cow interaction (except WC), age of cow x sex of calf interaction (only for WC), and covariates for B fraction of sire and cow, heterosis of cow and calf, and ELISA score. Random effects were sire (except for TO and CI), dam, and residual. Regression estimates of cow and calf traits on ELISA scores indicated that lower cow fertility (longer TO), lower ability of cows to maintain weight (negative WC), lower calf BWT, and lower calf WW205 were associated with higher cow ELISA scores. Further research on the effects of subclinical paratuberculosis in beef cattle at regional and national levels seems advisable considering the large potential economic cost of this disease. PMID:18998232

  8. Seed washing, exogenous application of gibberellic acid, and cold stratification enhance the germination of sweet cherry (Prunus avium L.) seed

    OpenAIRE

    Javanmard, T.; Zamani, Z; Keshavarz Afshar, R.; M. Hashemi; 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, the application of exogenous gibberellic acid (GA3), or cold stratification were evaluated for their ability to enhance the percentage and rate of seed germination. The results indicated that seed ...

  9. Rapid differentiation of Mycobacterium xenopi from mycobacteria of the Mycobacterium avium-intracellulare complex by pyrolysis mass spectrometry.

    OpenAIRE

    Sisson, P. R.; Freeman, R.; Magee, J G; Lightfoot, N F

    1992-01-01

    Thirty four cultures of slow growing, Tween-80 negative mycobacteria were analysed by pyrolysis mass spectrometry. The results showed that pyrolysis mass spectrometry could positively distinguish strains of Mycobacterium xenopi from those of the Mycobacterium avium-intracellulare (MAI) complex. Pyrolysis mass spectrometry may be a useful technique for the rapid characterisation of non-tuberculous mycobacteria in such clinical settings as their isolation from immunocompromised patients-for exa...

  10. Rapid diagnosis by buffy coat smear of disseminated Mycobacterium avium complex infection in patients with acquired immunodeficiency syndrome.

    OpenAIRE

    Nussbaum, J. M.; Dealist, C; Lewis, W; Heseltine, P N

    1990-01-01

    A smear of the buffy coat of peripheral blood for acid-fast bacilli was assessed for sensitivity and specificity in the diagnosis of disseminated Mycobacterium avium complex (MAC) infection in acquired immunodeficiency syndrome (AIDS) patients. Seventeen AIDS patients with blood cultures positive for MAC had simultaneous quantitative blood cultures and buffy coat smears performed, as did 4 patients later proven not to have disseminated MAC. The sensitivity of the buffy coat smear for the dete...

  11. Genetic Diversity of Mycobacterium avium Isolates Recovered from Clinical Samples and from the Environment: Molecular Characterization for Diagnostic Purposes▿

    OpenAIRE

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

    2008-01-01

    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...

  12. Characterization of sour (Prunus cerasus L.) and sweet cherry (Prunus avium L.) varieties with five isozyme systems

    OpenAIRE

    Morales Corts, Remedios; Rodrigues, Luciano MR; Ortiz, Jesús María; Pérez Sánches, Rodrigo

    2008-01-01

    Extracts from young leaves of nine sweet cherry (Prunus avium L.) and eight sour cherry (Prunus cerasus L.) varieties, located in the germplasm collection of the 'Direção Regional de Agricultura da Beira Interior' (Fundão, Portugal), were analysed for five isozyme systems in order to characterise these varieties and detect problems of synonymies and homonymies that frequently present. The sweet and sour cherry varieties analyzed showed low isoenzymatic polymorphism, being PGM and PGI the syst...

  13. Morphological indicators of the quality of one-year-old bare-root seedlings of wild cherry (Prunus avium L.)

    OpenAIRE

    Stjepanović Stefan; Ivetić Vladan

    2013-01-01

    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 s...

  14. Molecular and quantitative signatures of biparental inbreeding depression in the self-incompatible tree species Prunus avium

    OpenAIRE

    Jolivet, C.; Rogge, M.; Degen, B.

    2012-01-01

    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 pollin...

  15. Syndrome of selective IgM deficiency with severe T cell deficiency associated with disseminated cutaneous mycobacterium avium intracellulaire infection

    OpenAIRE

    Gharib, Asal; Louis, Ankmalika Gupta; Agrawal, Sudhanshu; Gupta, Sudhir

    2015-01-01

    Cutaneous non-disseminated, non-tuberculous mycobacterial infections have been reported in both immunocompetent and immunocompromised subjects. Systemic Mycobacterium avium intracellulaire (MAI) have been reported in non-HIV patients with Idiopathic CD4 lymphocytopenia. We report a comprehensive immunological analysis in syndrome of selective IgM deficiency and T lymphocytopenia (both CD4+ and CD8+) with disseminated cutaneous MAI infection. Naïve (TN) and Central memory (TCM) subsets of both...

  16. Factors affecting within orchard variability of nutrition,yield and quality of sweet cherry (Prunus avium L.)

    OpenAIRE

    Neilsen, Gerry H.; Neilsen, Denise; Herbert, Linda; Losso, Istvan; Rabie, Bill

    2009-01-01

    Continuous air temperature measurements were made during 2006-2008 at 30 different locations within a topographically complex orchard containing 10 yr old ‘Sweetheart’ sweet cherry (Prunus avium L.) on Mazzard rootstock in southern British Columbia. Yield, trunk cross-sectional area, fruit quality, and leaf and fruit mineral concentrations were measured throughout the study on adjacent trees at each location. Granier sap flow probes were installed on 10 nearby ‘Sweetheart’ trees to monitor ...

  17. 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.

  18. Ordeñando micobacterias del ganado? Impacto económico y en salud de Tuberculosis bovina y Paratuberculosis en Colombia

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    Jacobus H. de Waard

    2010-08-01

    Full Text Available La tuberculosis bovina y la paratuberculosis o enfermedad de Johne’s son enfermedades infecciosas en rumiantes, causadas por M. bovis y M. paratuberculosis respectivamente. La tuberculosis bovina es una enfermedad crónica de los rumiantes que guarda estrecha relación con la tuberculosis humana y provoca un deterioro progresivo del estado general de salud del animal, pérdida de peso, muy a menudo con tos y, a la larga, la muerte. La Paratuberculosis es una enfermedad infecto-contagiosa del tracto intestinal y se caracteriza por provocar enteritis granulomatosa crónica y progresiva, y diarreas sin respuesta a un tratamiento. Ambas enfermedades afectan especialmente los bovinos. Ambas bacterias son excretadas en la leche de los bovinos afectados, y por eso el consumo de leche cruda o queso fresco elaborado con leche cruda es la fuente de diseminación más importante de estas enfermedades tanto como entre los animales y al hombre. El impacto de estas enfermedades para la salud humana y la sanidad animal, la economía y el comercio es significativo y por esta razón están entre las enfermedades de declaración obligatoria a La Organización Mundial de Sanidad Animal (OIE (1.

  19. Abnormalities in the handling of intracellular bacteria in Crohn's disease: a link between infectious etiology and host genetic susceptibility.

    Science.gov (United States)

    Glasser, Anne-Lise; Darfeuille-Michaud, Arlette

    2008-01-01

    The etiology of Crohn's disease (CD) is still poorly understood, but recent advances have highlighted the importance of the innate immune system and the critical relationship between the gut flora and the intestinal mucosa. Several combinations of genetic factors predisposing to CD have been described, with the most significant replicable associations including genes for intracellular receptors of bacterial cell walls (NOD2/CARD15) and for bacterial clearance and antigen processing via autophagy (ATG16L1 and IRGM). One theoretical link between susceptibility genes NOD2/CARD15, ATG16L1, and IRGM is that CD is primarily induced by the presence of a dysfunctional immunological response to persistent infection by intracellular bacterial pathogens such as Mycobacterium avium subspecies paratuberculosis or adherent-invasive Escherichia coli, both first-rank candidates on the basis of host genetic susceptibility, which concerns impaired functions in the defense against intracellular bacteria. PMID:18726145

  20. Characterization of new IS elements and studies of their dispersion in two subspecies of Leifsonia xyli

    Directory of Open Access Journals (Sweden)

    Monteiro-Vitorello Claudia B

    2008-07-01

    Full Text Available Abstract Background Leifsonia xyli is a xylem-inhabiting bacterial species comprised of two subspecies: L. xyli subsp. xyli (Lxx and L. xyli subsp. cynodontis (Lxc. Lxx is the causal agent of ratoon stunting disease in sugarcane commercial fields and Lxc colonizes the xylem of several grasses causing either mild or no symptoms of disease. The completely sequenced genome of Lxx provided insights into its biology and pathogenicity. Since IS elements are largely reported as an important source of bacterial genome diversification and nothing is known about their role in chromosome architecture of L. xyli, a comparative analysis of Lxc and Lxx elements was performed. Results Sample sequencing of Lxc genome and comparative analysis with Lxx complete DNA sequence revealed a variable number of IS transposable elements acting upon genomic diversity. A detailed characterization of Lxc IS elements and a comparative review with IS elements of Lxx are presented. Each genome showed a unique set of elements although related to same IS families when considering features such as similarity among transposases, inverted and direct repeats, and element size. Most of the Lxc and Lxx IS families assigned were reported to maintain transposition at low levels using translation regulatory mechanisms, consistent with our in silico analysis. Some of the IS elements were found associated with rearrangements and specific regions of each genome. Differences were also found in the effect of IS elements upon insertion, although none of the elements were preferentially associated with gene disruption. A survey of transposases among genomes of Actinobacteria showed no correlation between phylogenetic relatedness and distribution of IS families. By using Southern hybridization, we suggested that diversification of Lxc isolates is also mediated by insertion sequences in probably recent events. Conclusion Collectively our data indicate that transposable elements are involved in