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Sample records for avium ssp paratuberculosis

  1. Seroprevalence of Mycobacterium avium SSP paratuberculosis ...

    African Journals Online (AJOL)

    This study aimed to determine the seroprevalence of antibodies for Mycobacterium avium subspecies paratuberculosis (MAP) in dairy cattle in the Jimma zone of Ethiopia in 2011. A random sample of 29 herds was selected, and all mature cattle within these herds had a blood sample taken. Serum was tested in duplicate, ...

  2. 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

  3. Immunogenicity of PtpA secreted during Mycobacterium avium ssp. paratuberculosis infection in cattle.

    Science.gov (United States)

    Bach, Eviatar; Raizman, Eran A; Vanderwal, Rich; Soto, Paolete; Chaffer, Marcelo; Keefe, Greg; Pogranichniy, Roman; Bach, Horacio

    2018-04-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne's disease. To survive within host macrophages, the pathogen secretes a battery of proteins to interfere with the immunological response of the host. One of these proteins is tyrosine phosphate A (PtpA), which has been identified as a secreted protein critical for survival of its close relative M. tuberculosis within infected macrophages. In this study, the immune response to recombinant PtpA used as an antigen was investigated in a cohort of ∼1000 cows infected with MAP compared to negative control animals using ELISA. The sera from MAP-infected cows had significantly higher levels of antibodies against PtpA when compared to uninfected cows. The data presented here indicate that the antibodies produced against PtpA are sensitive enough to detect infected animals before the appearance of the disease symptoms. The use of PtpA as an antigen can be developed as an early diagnostic test. Moreover, PtpA is a candidate antigen for detection of humoral immune responses in cows infected with MAP. Copyright © 2018 Elsevier B.V. All rights reserved.

  4. Survival of Mycobacterium avium ssp. paratuberculosis in yoghurt and in commercial fermented milk products containing probiotic cultures.

    Science.gov (United States)

    Van Brandt, L; Coudijzer, K; Herman, L; Michiels, C; Hendrickx, M; Vlaemynck, G

    2011-05-01

    To assess the survival of Mycobacterium avium ssp. paratuberculosis (MAP) in yoghurt and commercial fermented milk products containing probiotic strains. Whole and skimmed UHT milk artificially inoculated with MAP were used to manufacture yoghurt, using two different yoghurt starter cultures. Five commercial fermented milk products were inoculated with MAP. Two different MAP strains were studied. The survival of MAP in all products was monitored by culture over a 6-week storage period at 6°C. In yoghurt, MAP counts did not change appreciably during the storage period. Fat content and type of yoghurt starter culture had no consistent effect on the survival of MAP. In the fermented milk products, survival patterns varied but resulted in a 1·5 to ≥3·8 log reduction for the Niebüll strain and a 1·2-2·2 log reduction for the NIZO strain after 6 weeks, depending on the probiotic starters present in the product. MAP easily survived in yoghurt but MAP numbers decreased in fermented milk products containing probiotic cultures. The results contribute to the lack of knowledge on the behaviour of MAP in yoghurt and fermented milk products containing probiotic cultures. This knowledge is valuable in the context of the risk of MAP transmission to humans via yoghurt and the possible contribution of probiotic fermented milk products to the elimination of MAP. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  5. Assessment of the relative sensitivity of milk ELISA for detection of Mycobacterium avium ssp. paratuberculosis infectious dairy cows.

    Science.gov (United States)

    Laurin, Emilie L; Sanchez, Javier; Chaffer, Marcelo; McKenna, Shawn L B; Keefe, Greg P

    2017-01-01

    Milk ELISA are commonly used for detection of Mycobacterium avium ssp. paratuberculosis (MAP) antibodies in dairy cows, due to low cost and quick processing for large numbers of samples. However, low sensitivity and variations from host and environmental factors can impede detection of MAP antibodies at early disease stages. The objectives of our study were to assess the sensitivity of milk ELISA in comparison with fecal tests and to evaluate how detectable antibody concentrations in milk vary with changes in fecal shedding of MAP, cow age, cow parity, days in milk, and time of year. To compare the sensitivity of a commercial milk ELISA with solid and broth fecal culture and with fecal real-time PCR, a longitudinal study was performed for the identification of MAP-infectious animals as determined by prior fecal testing for MAP shedding. In addition, associations between variation in milk MAP ELISA score and changes in fecal MAP shedding, host age, days in milk, and season were evaluated. Monthly milk and fecal samples were collected over 1 yr from 46 cows that were previously shedding MAP in their feces. Sensitivity of milk ELISA was 29.9% (95% CI: 24.8 to 35.1%), compared with 46.7% (40.7 to 52.7%) for fecal solid culture, 55.0% (49.3 to 60.7%) for fecal broth culture, and 78.4% (73.3 to 83.1%) for fecal direct real-time PCR. The effect of stage of lactation could not be separated from the effect of season, with increased milk ELISA scores at greater days in milk in winter. However, unpredictable monthly variations in results were observed among the 3 assays for individual cow testing, which highlights the importance of identifying patterns in pathogen and antibody detection over time in MAP-positive herds. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  6. 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.

  7. Mycobacterium avium ssp. paratuberculosis detection in animals, food, water and other sources or vehicles of human exposure: A scoping review of the existing evidence.

    Science.gov (United States)

    Waddell, Lisa; Rajić, Andrijana; Stärk, Katharina; McEwen, Scott A

    2016-09-15

    Mycobacterium avium ssp. paratuberculosis is the etiologic agent of Johne's disease in ruminants and is hypothesized to be an infectious cause of Crohn's disease, as well as some other human diseases. Due to key knowledge gaps, the potential public health impact of M. paratuberculosis is unknown. This scoping review aims to identify and characterised the evidence on potential sources and vehicles of M. paratuberculosis exposure for humans to better understand how exposure is likely to occur. Evidence from 255 primary research papers is summarized; most examined the prevalence or concentration of M. paratuberculosis in animals (farmed domestic, pets and wildlife) (n=148), food for human consumption (62) (milk, dairy, meat, infant formula) or water (drinking and recreational) and the environment (farm, pasture and areas affected by runoff water) (20). The majority of this research has been published since 2000 (Figure- abstract). Nine case-control studies examining risk factors for Crohn's disease highlighted significant associations with the consumption of processed meats and cheese, while direct contact with ruminants, high risk occupations (farmer, veterinarian), milk consumption and water source were factors not associated with the disease and/or M. paratuberculosis exposure status. Molecular epidemiology studies demonstrated strain-sharing between species. Produce and seafood were the only previously suggested sources of human exposure for which there was no supporting evidence identified in this scoping review. The results of this review indicate that ruminant populations from around the globe are infected with M. paratuberculosis and many non-ruminant species have also been found to carry or be infected with M. paratuberculosis. Several potential sources for human exposure to M. paratuberculosis were identified; however there remain important gaps in quantitative information on the prevalence and concentration of M. paratuberculosis in contaminated sources of

  8. 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. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  9. Effect of feeding heat-treated colostrum on risk for infection with Mycobacterium avium ssp. paratuberculosis, milk production, and longevity in Holstein dairy cows.

    Science.gov (United States)

    Godden, S M; Wells, S; Donahue, M; Stabel, J; Oakes, J M; Sreevatsan, S; Fetrow, J

    2015-08-01

    In summer 2007, a randomized controlled field trial was initiated on 6 large Midwest commercial dairy farms to investigate the effect of feeding heat-treated (HT) colostrum on transmission of Mycobacterium avium ssp. paratuberculosis (MAP) and on future milk production and longevity within the herd. On each farm, colostrum was collected daily from fresh cows, pooled, divided into 2 aliquots, and then 1 aliquot was heat-treated in a commercial batch pasteurizer at 60°C for 60min. A sample from each batch of colostrum was collected for PCR testing (MAP-positive vs. MAP-negative). Newborn heifer calves were removed from the dam within 30 to 60min of birth and systematically assigned to be fed 3.8 L of either fresh (FR; n=434) or heat-treated (HT; n=490) colostrum within 2h of birth. After reaching adulthood (>2 yr old), study animals were tested once annually for 3 yr (2010, 2011, 2012) for infection with MAP using serum ELISA and fecal culture. Lactation records describing milk production data and death or culling events were collected during the 3-yr testing period. Multivariable model logistic and linear regression was used to investigate the effect of feeding HT colostrum on risk for testing positive to MAP during the 3-yr testing period (positive/negative; logistic regression) and on first and second lactation milk yield (kg/cow; linear regression), respectively. Cox proportional hazards regression was used to investigate the effect of feeding HT colostrum on risk and time to removal from the herd. Fifteen percent of all study animals were fed PCR-positive colostrum. By the end of the 3-yr testing period, no difference was noted in the proportion of animals testing positive for MAP, with either serum ELISA or fecal culture, when comparing the HT group (10.5%) versus the FR group (8.1%). There was no effect of treatment on first- (HT=11.797kg; FR=11,671kg) or second-lactation (HT=11,013kg; FR=11,235kg) milk production. The proportion of cows leaving the herd by

  10. 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.

    Paratuberculosis in ruminants is caused by an infection with Mycobacterium avium subspecies paratuberculosis (MAP) and is a chronic disease characterized by granulomatous enteritis. Available vaccines against paratuberculosis consist of variations of whole bacteria with adjuvant showing various...

  11. Reduction of Mycobacterium avium ssp. paratuberculosis in colostrum: Development and validation of 2 methods, one based on curdling and one based on centrifugation.

    Science.gov (United States)

    Verhegghe, M; Rasschaert, G; Herman, L; Goossens, K; Vandaele, L; De Bleecker, K; Vlaemynck, G; Heyndrickx, M; De Block, J

    2017-05-01

    The aim of this study was to develop and validate 2 protocols (for use on-farm and at a central location) for the reduction of Mycobacterium avium ssp. paratuberculosis (MAP) in colostrum while preserving beneficial immunoglobulins (IgG). The on-farm protocol was based on curdling of the colostrum, where the IgG remain in the whey and the MAP bacteria are trapped in the curd. First, the colostrum was diluted with water (2 volumes colostrum to 1 volume water) and 2% rennet was added. After incubation (1 h at 32°C), the curd was cut and incubated again, after which whey and curd were separated using a cheesecloth. The curd was removed and milk powder was added to the whey. Approximately 1 log reduction in MAP counts was achieved. A reduction in total proteins and IgG was observed due to initial dilution of the colostrum. After curd formation, more than 95% of the immunoglobulins remained in the whey fraction. The semi-industrial protocol was based on centrifugation, which causes MAP to precipitate, while the IgG remain in the supernatant. This protocol was first developed in the laboratory. The colostrum was diluted with skimmed colostrum (2 volumes colostrum to 1 volume skimmed colostrum), then skimmed and centrifuged (at 15,600 × g for 30 min at room temperature). We observed on average 1.5 log reduction in the MAP counts and a limited reduction in proteins and IgG in the supernatant. To obtain a semi-industrial protocol, dairy pilot appliances were evaluated and the following changes were applied to the protocol: after 2:1 dilution as above, the colostrum was skimmed and subsequently clarified, after which the cream was heat treated and added to the supernatant. To investigate the effect of the colostrum treatment on the nutritional value and palatability of the colostrum and the IgG transfer, an animal experiment was conducted with 24 calves. Six received the dam's colostrum, 6 were given untreated purchased colostrum (control), and 2 groups of 6 calves

  12. Rapid detection of Mycobacterium avium subsp. paratuberculosis ...

    African Journals Online (AJOL)

    Therefore, alternative diagnostic tests such as PCR, are needed for quick detection of infected animals. In this study, the conventional enrichment and isolation procedure and two IS900-based PCR methods for detection of Mycobactrium avium subsp. paratuberculosis in clinical samples from zoo animals and cattle were ...

  13. Comparative risk assessment for new cow-level Mycobacterium avium ssp. paratuberculosis infections between 3 dairy production types: Organic, conventional, and conventional-grazing systems.

    Science.gov (United States)

    Beaver, A; Ruegg, P L; Gröhn, Y T; Schukken, Y H

    2016-12-01

    Johne's disease, a granulomatous enteritis of ruminant animals, is a hidden threat on dairy farms, adversely affecting animal welfare as well as herd productivity. Control programs in the United States advocate for specific management practices to temper the spread of the causal organism (Mycobacterium avium ssp. paratuberculosis, MAP), such as improving calving area hygiene and limiting introduction of replacement stock with unknown infection status. A need remains for direct exploration of Johne's disease prevention strategies in the United States with respect to production type. Alongside the growing demand for organic products, the safety of organic dairy practices with respect to MAP control is warranted. Further, conventional herds for which organic practices such as pasture grazing are used should be situated within the risk spectrum. We developed a risk assessment model using the US Voluntary Bovine Johne's Disease Control Program as a framework, with the goal of evaluating the risk of new cow-level MAP infections. A total of 292 organic and conventional farms in 3 states were surveyed on management practices, and an overall analysis was conducted in which each farm was first scored on individual practices using a range of "no risk" to "high risk," according to the literature. The sum of all risk factors was then analyzed to quantify and compare the risk burden for each production type. Organic herds received higher overall risk scores compared with both conventional grazing and nongrazing subtypes. To identify which factors contributed to the overall increased risk for organic herds, the management practices were categorized and evaluated by logistic regression. We determined that the increased risk incurred by organic herds was predominantly due to decisions made in the calving area and preweaned calf group. However, although certain individual risk factors related to calf management are commonly involved in prevention strategies (e.g., cow

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

    NARCIS (Netherlands)

    Weber, M.F.

    2009-01-01

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

  15. Environmental contamination with Mycobacterium avium subspecies paratuberculosis within and around a dairy barn under experimental conditions

    NARCIS (Netherlands)

    Eisenberg, S.W.F.; Nielen, M.; Hoeboer, J.; Rutten, V.P.; Heederik, D.; Koets, A.P.

    2012-01-01

    To establish environmental contamination in and around a dairy barn, cows shedding Mycobacterium avium ssp. paratuberculosis (MAP) were housed in a freestall barn. Fecal samples were collected 15 times at 3-wk intervals, and samples of all animals were cultured by using the Trek Diagnostic Systems

  16. 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.

  17. Search for Mycobacterium avium Subspecies paratuberculosis Antigens for the Diagnosis of Paratuberculosis

    NARCIS (Netherlands)

    Mon, M.L.; Vale, M.; Baschetti, G.; Alvarado Pinedo, F.; Gioffre, A.; Traveria, G.; Willemsen, P.; Bakker, D.; Romano, M.I.

    2012-01-01

    The aim of this study was to evaluate a wide panel of antigens of Mycobacterium avium subsp. paratuberculosis (MAP) to select candidates for the diagnosis of paratuberculosis (PTB). A total of 54 recombinant proteins were spotted onto nitrocellulose membranes and exposed to sera from animals with

  18. 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....

  19. 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...

  20. Short communication: Recovery of viable Mycobacterium avium subspecies paratuberculosis from retail pasteurized whole milk in Brazil.

    Science.gov (United States)

    Carvalho, I A; Pietralonga, P A G; Schwarz, D G G; Faria, A C S; Moreira, M A S

    2012-12-01

    Mycobacterium avium ssp. paratuberculosis (MAP) is the etiological agent of paratuberculosis, a chronic granulomatous enteritis that affects all ruminants worldwide. Some researchers have indicated a possible role of MAP in Crohn's disease. Despite extensive research and large and important advances in the past few decades, the etiology of Crohn's disease remains indefinite. The most probable transmission route of MAP from animals to humans is milk and dairy products. Mycobacterium avium ssp. paratuberculosis has already been detected in milk samples worldwide, and some studies have reported that MAP is resistant to pasteurization. In Brazil, MAP has been reported in raw milk samples; however, Brazilian retail pasteurized milk has not yet been tested for viable MAP. The aim of this study was to investigate MAP in pasteurized milk in the region of Viçosa (Minas Gerais, Brazil). Thirty-seven samples were collected and processed for culture of MAP. One colony similar to MAP was observed and confirmed by IS900-nested PCR and sequencing. Analysis revealed 97 to 99% identity with the MAP K-10 strain. This study is the first report of the presence of MAP in retail pasteurized whole milk in Brazil. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  1. 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...

  2. 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...

  3. Mycobacterium avium subspecies paratuberculosis recombinant proteins modulate antimycobacterial functions of bovine macrophages

    Science.gov (United States)

    It has been shown that Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) activates the Mitogen Activated Protein Kinase (MAPK) p38 pathway, yet it is unclear which components of M. paratuberculosis are involved in the process. Therefore, a set of 42 M. paratuberculosis recombinan...

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

    Directory of Open Access Journals (Sweden)

    Chung-Yi eHsu

    2011-12-01

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

  5. Environmental Mycobacterium avium subsp. paratuberculosis Hosted by Free-Living Amoebae

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    Ascel Samba-Louaka

    2018-02-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis is responsible for paratuberculosis in animals. This disease, leading to an inflammation of the gastrointestinal tract, has a high impact on animal health and an important economic burden. The environmental life cycle of M. avium subsp. paratuberculosis is poorly understood and several studies suggest that free-living amoebae (FLA might be a potential environmental host. FLA are protozoa found in water and soil that are described as reservoirs of pathogenic and non-pathogenic bacteria in the environment. Indeed, bacteria able to survive within these amoebae would survive phagocytosis from immune cells. In this study, we assessed the in vitro interactions between several strains of M. avium subsp. paratuberculosis and Acanthamoeba castellanii. The results indicate that the bacteria were able to grow within the amoeba and that they can survive for several days within their host. To explore the presence of M. avium subsp. paratuberculosis in environmental amoebae, we sampled water from farms positive for paratuberculosis. A M. avium subsp. paratuberculosis strain was detected within an environmental amoeba identified as related to the poorly described Rosculus genus. The bacterial strain was genotyped, showing that it was similar to previous infectious strains isolated from cattle. In conclusion, we described that various M. avium subsp. paratuberculosis strains were able to grow within amoebae and that these bacteria could be found on farm within amoebae isolated from the cattle environment. It validates that infected amoebae might be a reservoir and vector for the transmission of M. avium subsp. paratuberculosis.

  6. Environmental Mycobacterium avium subsp. paratuberculosis hosted by free-living amoebae

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis is responsible for paratuberculosis in animals. This disease, leading to an inflammation of the gastrointestinal tract, has a high impact on animal health and an important economic burden. The environmental life cycle of Mycobacterium avium subsp. paratube...

  7. Molecular biology techniques as a tool for detection and characterisation of Mycobacterium avium subsp. paratuberculosis

    OpenAIRE

    Englund, Stina

    2002-01-01

    Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) is the causative agent of paratuberculosis, also known as Johne’s disease, a chronic intestinal infection in cattle and other ruminants. Paratuberculosis is characterised by diarrhea and weight loss that occurs after a period of a few months up to several years without any clinical signs. The considerable economic losses to dairy and beef cattle producers are caused by reduced milk production and poor reproduction performance i...

  8. A Mycobacterium avium subsp. paratuberculosis predicted serine protease is associated with acid stress and intraphagosomal survival

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is an intracellular pathogen that persists inside host macrophages despite severe oxidative stress and nutrient deprivation. Intrabacterial pH homeostasis is vital to pathogenic mycobacteria to preserve cellular biological processes and stability of ...

  9. Composition and potency characterization of Mycobacterium avium subsp. paratuberculosis purified protein derivatives

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) purified protein derivatives (PPDs) are immunologic reagents prepared from cultured filtrates of the type strain ATCC 19698. Traditional production consists of floating culture incubation at 37oC, organism inactivation by autoclaving, coarse filtrat...

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

    DEFF Research Database (Denmark)

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

    2015-01-01

    Background: Mycobacterium avium subsp. paratuberculosis (MAP) infections in cattle are generally challenging to detect and cost-effective test strategies are consequently difficult to identify. MAP-specific antibody ELISAs for milk and serum are relatively inexpensive, but their utility is influe......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...

  11. Persistence of Mycobacterium avium subsp. paratuberculosis at a Farm-Scale Biogas Plant Supplied with Manure from Paratuberculosis-Affected Dairy Cattle▿

    Science.gov (United States)

    Slana, I.; Pribylova, R.; Kralova, A.; Pavlik, I.

    2011-01-01

    In this study, products from all steps of anaerobic digestion at a farm-scale biogas plant supplied with manure from paratuberculosis-affected dairy cattle were examined and quantified for the presence of the causal agent of paratuberculosis, Mycobacterium avium subsp. paratuberculosis, using culture and quantitative real-time PCR (qPCR). Viable M. avium subsp. paratuberculosis cells were detected using culture in fermentors for up to 2 months; the presence of M. avium subsp. paratuberculosis DNA (101 cells/g) was demonstrated in all anaerobic fermentors and digestate 16 months after initiation of work at a biogas plant, using IS900 qPCR. F57 qPCR was able to detect M. avium subsp. paratuberculosis DNA (102 cells/g) at up to 12 months. According to these results, a fermentation process that extended beyond 2 months removed all viable M. avium subsp. paratuberculosis cells and therefore rendered its product M. avium subsp. paratuberculosis free. However, M. avium subsp. paratuberculosis DNA was found during all the examined periods (more than 1 year), which could be explained by either residual DNA being released from dead cells or by the presence of viable cells whose amount was under the limit of cultivability. As the latter hypothesis cannot be excluded, the safety of the final products of digestion used for fertilization or animal bedding cannot be defined, and further investigation is necessary to confirm or refute this risk. PMID:21398476

  12. Vaccination with peptides of Mycobacterium avium subsp. paratuberculosis (MAP) reduces MAP burden of infected goats

    DEFF Research Database (Denmark)

    Melvang, Heidi Mikkelsen; Hassan, Sufia Butt; Thakur, Aneesh

    Mycobacterium avium subsp. paratuberculosis (Map) is the cause of paratuberculosis, a chronic enteritis of ruminants that is widespread worldwide. We investigated the effect of post-exposure vaccination with Map specific peptides in a goat model aiming at developing a Map vaccine that will neither...

  13. 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

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

    DEFF Research Database (Denmark)

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

    2009-01-01

    Paratuberculosis is a chronic infection of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). It is possible to detect infection with paratuberculosis at different stages of disease by means of various diagnostic test strategies. The objective of the present study was to evalu......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...

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

    DEFF Research Database (Denmark)

    Afzal, Mamuna; Abidi, Soad; Mikkelsen, Heidi

    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 Löwen......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......, 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....

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

    Science.gov (United States)

    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 clin...

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

    DEFF Research Database (Denmark)

    Giese, Steen Bjørck; Ahrens, Peter

    1999-01-01

    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......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......-negative on intestinal mucosa, but culture-positive in milk, and both faeces and milk were negative in culture and PCR from 2 cows. In conclusion the presence of M. a. paratuberculosis could be detected in raw milk by PCR but cultivation of milk was more sensitive in detecting the organism....

  18. Impact of salt concentration on persistence of Mycobacterium avium subsp. paratuberculosis in Iranian UF white cheese

    Directory of Open Access Journals (Sweden)

    Sh Hanifian

    2013-02-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (Mycobacterium paratuberculosis is considered as a potential significant public health threat due to its possible association with Crohn’s disease in humans. This is a study aimed to investigate the effect of different salt concentrations on survival of Mycobacterium paratuberculosis during ripening and storage of Iranian ultra-filtrate-white cheese (IUFWC. For this purpose, retentate was inoculated with 2 Log cfu/g of Mycobacterium paratuberculosis. Afterwards, model cheeses were prepared with 2%, 3% and 4% of salt. Quantity of Mycobacterium paratuberculosis was estimated throughout the ripening and storage of IUFWC using F57-quantitative real time PCR (F57-qPCR and culture assay. Along with, the populations of lactic acid bacteria as well as physicochemical properties of cheese samples were determined. According to the results, at the early stage of storage period (1 to 30 days the number of Mycobacterium paratuberculosis was almost constant; however, it was decreased significantly (p

  19. Heat shock protein 70 and bovine paratuberculosis

    NARCIS (Netherlands)

    Langelaar, Merel Frederiek Moira

    2005-01-01

    Paratuberculosis, or Johne’s disease, is a chronic infection of the small intestine of cattle caused by Mycobacterium avium ssp. paratuberculosis (M.a.p.) , leading to an incurable form of protein loosing enteropathy. Young animals are orally infected with M.a.p., which resides and replicates in

  20. The effect of Protexin on prevention of ileocecal infection by Mycobacterium avium subspecies paratuberculosis in dairy calves.

    Science.gov (United States)

    Badiei, A; Moosakhani, F; Hamidi, A; Sami, M

    2013-10-01

    The objective of this study was to evaluate the effect of Protexin (Probiotics International Ltd., South Petherton, UK) in the prevention of ileocecal infection by Mycobacterium avium ssp. paratuberculosis (MAP) in dairy calves in the field situation. Forty Holstein bull calves whose dams were paratuberculosis negative (confirmed by serum ELISA test and fecal nested PCR) were randomly selected in 2 groups. All calves were fed raw milk collected from the bulk tank in a paratuberculosis-infected dairy farm, which was confirmed by PCR. The treatment group (20 calves) was given 2 g of Protexin from birth until weaning (90 d). The control group (20 calves) did not consume Protexin. The calves were culled at 12 mo of age and the ileocecal lymph nodes were sampled. The lymph nodes were tested by nested PCR to evaluate MAP infection. In the treatment group, 2 out of 20 calf (10%) ileoceca were infected by MAP, whereas in the control group, 8 out of 20 calf (40%) ileoceca were infected by MAP. A significant difference existed between ileocecal infection by MAP in treatment and control groups. Thus, Protexin showed a significant effect in decreasing the ileocecal infection by MAP. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Mariana Noelia Viale

    2014-01-01

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

  2. Inferring biomarkers for Mycobacterium avium subsp. paratuberculosis infection and disease progression using experimental data

    Science.gov (United States)

    Available diagnostic assays for Mycobacterium avium subsp paratuberculosis (MAP) have poor sensitivities and cannot detect early stages of the infection, therefore, there is need to find new diagnostic markers for early infection detection and disease stages. We analyzed longitudinal IFN- gamma, ELI...

  3. 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...

  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. Transcriptional profiling of ileocecal valve of Holstein dairy cows infected with mycobacterium avium subsp. paratuberculosis

    Science.gov (United States)

    Johne’s disease is a chronic infection of the small intestine caused by Mycobacterium avium subspecies paratuberculosis (MAP), an intracellular bacterium. The events of pathogen survival within the host cell(s), chronic inflammation and the progression from asymptomatic subclinical stage to an advan...

  6. Lymphoproliferative and gamma interferon responses to stress-regulated Mycobacterium avium subsp. paratuberculosis recombinant proteins

    Science.gov (United States)

    Johne’s disease in ruminants is a chronic infection of the intestines caused by Mycobacterium avium subsp. paratuberculosis. Economic losses associated with Johne’s disease arise due to premature culling, reduced production of milk and wool and mortalities. The disease is characterised by a long inc...

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

    Science.gov (United States)

    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...

  8. 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...

  9. Environmental contamination with Mycobacterium avium subsp. paratuberculosis in endemically infected dairy herds

    Science.gov (United States)

    Environmental contamination with Mycobacterium avium subsp. paratuberculosis (MAP) is thought to be the primary source of infection for dairy cattle. The exact link between fecal shedding of MAP by individual cows and environmental contamination levels at the herd level was explored with a cross-se...

  10. 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.

  11. Molecular characterization of Mycobacterium avium subspecies paratuberculosis Type II and III isolates by a combination of MIRU-VNTR loci

    OpenAIRE

    2009-01-01

    Abstract Mycobacterial interspersed repetitive units and variable number tandem repeats typing (MIRU-VNTR) is a useful technique that has been recently applied to characterize members of the Mycobacterium avium complex (MAC). The aim of this study was to examine the genetic variability among a collection of Spanish Mycobacterium avium subspecies paratuberculosis (M. a. paratuberculosis) isolates with a combination of MIRU-VNTR loci. For this purpose we tested six MIRU-VNTR loci (MI...

  12. 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...

  13. Economic analysis of Mycobacterium avium subspecies paratuberculosis vaccines in dairy herds.

    Science.gov (United States)

    Cho, J; Tauer, L W; Schukken, Y H; Gómez, M I; Smith, R L; Lu, Z; Grohn, Y T

    2012-04-01

    Johne's disease, or paratuberculosis, is a chronic infectious enteric disease of ruminants, caused by infection with Mycobacterium avium ssp. paratuberculosis (MAP). Given the absence of a fail-safe method of prevention or a cure, Johne's disease can inflict significant economic loss on the US dairy industry, with an estimated annual cost of over $200 million. Currently available MAP control strategies include management measures to improve hygiene, culling MAP serologic- or fecal-positive adult cows, and vaccination. Although the 2 first control strategies have been reported to be effective in reducing the incidence of MAP infection, the changes in herd management needed to conduct these control strategies require significant effort on the part of the dairy producer. On the other hand, vaccination is relatively simple to apply and requires minor changes in herd management. Despite these advantages, only 5% of US dairy operations use vaccination to control MAP. This low level of adoption of this technology is due to limited information on its cost-effectiveness and efficacy and some important inherent drawbacks associated with current MAP vaccines. This study investigates the epidemiological effect and economic values of MAP vaccines in various stages of development. We create scenarios for the potential epidemiological effects of MAP vaccines, and then estimate economically justifiable monetary values at which vaccines become economically beneficial to dairy producers such that a net present value (NPV) of a farm's net cash flow can be higher than the NPV of a farm using no control or alternative nonvaccine controls. Any vaccination with either low or high efficacy considered in this study yielded a higher NPV compared with a no MAP control. Moreover, high-efficacy vaccines generated an even higher NPV compared with alternative controls, making vaccination economically attractive. Two high-efficacy vaccines were particularly effective in MAP control and NPV

  14. Mycobacterium avium subsp. paratuberculosis: presencia en los alimentos y su relación con la enfermedad de Crohn Mycobacterium avium subsp. paratuberculosis in food and its relationship with Crohn's disease

    Directory of Open Access Journals (Sweden)

    K. Cirone

    2007-03-01

    Full Text Available La paratuberculosis o enfermedad de Johne es una enteritis crónica producida por Mycobacterium avium subsp. paratuberculosis, que afecta a bovinos y a otras especies. En la Argentina se ha caracterizado en rodeos bovinos y de ciervos, con aislamientos tipificados en distintos patrones genéticos. M. avium subsp. paratuberculosis ha sido vinculado en humanos con una inflamación crónica del intestino, denominada enfermedad de Crohn. Existen evidencias clínicas y experimentales que relacionan a M. avium subsp. paratuberculosis con la enfermedad en el humano, mediante su detección por PCR y por cultivo a partir de biopsias de órganos, de leche materna y de sangre de pacientes afectados. La leche y sus subproductos serían posibles fuentes de infección y se ha sugerido que M. avium subsp. paratuberculosis resistiría las condiciones de pasteurización. Diversos trabajos de investigación demostraron que esta micobacteria podría estar presente en leches comercializadas en diversos países, como Reino Unido, Estados Unidos, República Checa, y también en la Argentina. La presencia de M. avium subsp. paratuberculosis en productos lácteos y agua de consumo ha sido relacionada con la resistencia del microorganismo tanto a los procesos de elaboración como a los factores climáticos adversos, lo que enfatiza el rol de los alimentos y del agua como vías de transmisión al humano. Las investigaciones en curso podrían ratificar el riesgo y las implicancias de la exposición del humano a M. avium subsp. paratuberculosis a través de los alimentos y del agua contaminados, para determinar la importancia de la paratuberculosis como enfermedad zoonótica.Paratuberculosis or Johne's disease is a chronic enteritis of the cattle and other small ruminant animals caused by Mycobacterium avium subsp. paratuberculosis. In Argentina, the strains were characterized in beef and dairy cattle and deer in different genetic patterns by molecular tools. M. avium

  15. Fate of Mycobacterium avium subsp. paratuberculosis after application of contaminated dairy cattle manure to agricultural soils.

    Science.gov (United States)

    Salgado, M; Collins, M T; Salazar, F; Kruze, J; Bölske, G; Söderlund, R; Juste, R; Sevilla, I A; Biet, F; Troncoso, F; Alfaro, M

    2011-03-01

    Details regarding the fate of Mycobacterium avium subsp. paratuberculosis (basonym, Mycobacterium paratuberculosis) after manure application on grassland are unknown. To evaluate this, intact soil columns were collected in plastic pipes (lysimeters) and placed under controlled conditions to test the effect of a loamy or sandy soil composition and the amount of rainfall on the fate of M. paratuberculosis applied to the soil surface with manure slurry. The experiment was organized as a randomized design with two factors and three replicates. M. paratuberculosis-contaminated manure was spread on the top of the 90-cm soil columns. After weekly simulated rainfall applications, water drainage samples (leachates) were collected from the base of each lysimeter and cultured for M. paratuberculosis using Bactec MGIT ParaTB medium and supplements. Grass was harvested, quantified, and tested from each lysimeter soil surface. The identity of all probable M. paratuberculosis isolates was confirmed by PCR for IS900 and F57 genetic elements. There was a lag time of 2 months after each treatment before M. paratuberculosis was found in leachates. The greatest proportions of M. paratuberculosis-positive leachates were from sandy-soil lysimeters in the manure-treated group receiving the equivalent of 1,000 mm annual rainfall. Under the higher rainfall regimen (2,000 mm/year), M. paratuberculosis was detected more often from lysimeters with loamy soil than sandy soil. Among all lysimeters, M. paratuberculosis was detected more often in grass clippings than in lysimeter leachates. At the end of the trial, lysimeters were disassembled and soil cultured at different depths, and we found that M. paratuberculosis was recovered only from the uppermost levels of the soil columns in the treated group. Factors associated with M. paratuberculosis presence in leachates were soil type and soil pH (P soils (faster through sandy soil) and tends to remain on grass and in the upper layers of pasture

  16. 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).

  17. Assessing virulence of vaccine strains of Mycobacterium avium subspecies paratuberculosis in a calf model.

    Science.gov (United States)

    Watkins, Craig; Schock, Alex; May, Linda; Denham, Susan; Sales, Jill; Welch, Louise; Sharp, J Michael; Stevenson, Karen

    2010-11-20

    The purpose of this investigation was to characterise the virulence of two Mycobacterium avium subspecies paratuberculosis (M.a. paratuberculosis) vaccine strains and compare them with a recent virulent isolate in new born calves over a time course of 8 months post-inoculation. Paratuberculosis-free new born calves were inoculated orally with either a vaccine strain (2e or 316F) or a wild type strain (F13) of M.a. paratuberculosis. Blood and faecal samples were collected throughout the experiment to analyse immune responses to infection and assess faecal shedding of M.a. paratuberculosis. Tissue samples were taken at post-mortem for histological examination and bacteriological culture. Cell-mediated immune responses were measured using a Bovigam (CSL) interferon-gamma assay. At 20 weeks post-inoculation there was a significant increase in the cell-mediated immune responses in calves infected with the wild type strain relative to the two vaccine strains. Acid fast bacteria were detected in the faeces of calves in all three groups between 4 and 8 weeks post-inoculation. Histopathology was unrewarding in all three groups. M.a. paratuberculosis was recovered only from tissues of calves inoculated with the wild type strain. Therefore, it appeared that the vaccine strains used in this study had reduced virulence. Identifying the genes responsible for pathogenesis observed in the wild type isolate and reduced or inactive in these vaccine isolates may offer a valuable resource for improving our knowledge of pathogenesis and permit the development of improved diagnostic reagents and vaccines for the control of M.a. paratuberculosis in livestock. Copyright © 2010 Elsevier B.V. All rights reserved.

  18. Molecular epidemiology of Types I/III strains of Mycobacterium avium subspecies paratuberculosis isolated from goats and cattle.

    Science.gov (United States)

    de Juan, L; Alvarez, J; Aranaz, A; Rodríguez, A; Romero, B; Bezos, J; Mateos, A; Domínguez, L

    2006-06-15

    Molecular characterization of Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis) isolates classifies them into three groups: cattle or Type II, sheep or Type I, and intermediate or Type III. To avoid problems associated with characterization of extremely slow growth strains, PCR-based techniques that divide the M. a. paratuberculosis strains in two main groups (cattle or Type II, and sheep or Types I/III) can be performed. The objectives of this study were to characterize the M. a. paratuberculosis isolates identified by different PCR-based tests (IS1311-PCR and restriction endonuclease analysis, PCR test based on a DNA sequence difference, and a PCR aimed at three Type I-specific loci), and to determine the clinical and epidemiological implications of Types I/III M. a. paratuberculosis strains in livestock. One hundred and fifty-eight M. a. paratuberculosis strains from domestic ruminants were analyzed. One hundred and six M. a. paratuberculosis isolates (61 from goats and 45 from cattle) were classified as Type II strains; and 52 (29 from cows, 20 from goats, and three from sheep) were included in the Types I/III. The Types I/III M. a. paratuberculosis strains were associated to Spanish native breeds. The majority of these animals had not been in direct or indirect contact with sheep flocks infected with M. a. paratuberculosis. This fact should be taken into account when implementing paratuberculosis control programs.

  19. 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

    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......Paratuberculosis is a chronic intestinal infection in ruminants, caused by Mycobacterium avium subspecies paratuberculosis (Map). To study the role of host genetics in disease susceptibility, the Toll-like receptor 2 (TLR2) gene, selected based on its potential role in immunity to mycobacterial...... paratuberculosis-infected animals and 12 age-matched healthy herd mates revealed 21 different SNP. The TLR2-1903 T/C SNP was significantly associated with resistance to Map. This and four additional TLR2 SNP were studied in a subsequent observational field study with 553 cows from farms with paratuberculosis...

  20. Divergent cellular responses during asymptomatic subclinical and clinical states of disease in cows naturally infected with Mycobacterium avium subsp. paratuberculosis

    Science.gov (United States)

    Infection of the host with Mycobacterium avium subsp. paratuberculosis (MAP) results in a chronic and progressive enteritis that traverses both subclinical and clinical stages. The mechanism(s) for the shift from asymptomatic subclinical disease state to advanced clinical disease are not fully under...

  1. Analysis of Mycobacterium avium subsp. paratuberculosis mutant libraries reveals loci-dependent transcription biases and strategies to novel mutant discovery

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne’s disease in ruminants and it has been implicated as a cause of Crohn’s disease in humans. The generation of comprehensive random mutant banks by transposon mutagenesis is a fundamental wide genomic technology utilized...

  2. Analysis of Mycobacterium avium subspecies paratuberculosis mutant libraries reveals loci-dependent transposition biases and strategies to novel mutant discovery

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP), the etiologic agent of Johne’s disease, is one of the most important bacterial pathogens in ruminants. The lack of efficacious control measures demands a thorough understanding of MAP pathogenesis to develop new vaccines and diagnostic tests. The ge...

  3. Antigenicity of recombinant maltose binding protein-Mycobacterium avium subsp. paratuberculosis fusion proteins with and without factor Xa cleaving

    Science.gov (United States)

    Mycobacterium avium subsp paratuberculosis (MAP) causes Johne’s disease (JD) in ruminants. Proteomic studies have shown that MAP expresses certain proteins when exposed to in vitro physiological stress conditions similar to the conditions experienced within a host during natural infection. Such prot...

  4. Evaluation of eight live attenuated vaccine candidates for protection against challenge with virulent Mycobacterium avium subspecies paratuberculosis in mice

    Science.gov (United States)

    Johne’s disease is caused by Mycobacterium avium subsp. paratuberculosis (MAP), which results in serious economic losses worldwide in farmed livestock such as cattle, sheep and goats. To control this disease, an effective vaccine with minimal adverse effects is needed. In order to identify a live va...

  5. Evaluation of two mutants of Mycobacterium avium subsp. paratuberculosis as candidates for a live attenuated vaccine for Johne's disease

    Science.gov (United States)

    Efforts to control Johne’s disease (JD), caused by Mycobacterium avium subsp. paratuberculosis (Map), has been difficult because of a lack of an effective vaccine. To address this problem we examined the potential of targeted gene disruption as a method to develop candidate vaccines with impaired c...

  6. UV Light Inactivation of Mycobacterium avium subsp. paratuberculosis in Milk as Assessed by FASTPlaqueTB Phage Assay and Culture▿

    Science.gov (United States)

    Altic, Leslie C.; Rowe, Michael T.; Grant, Irene R.

    2007-01-01

    UV light inactivation of Mycobacterium avium subsp. paratuberculosis in Middlebrook 7H9 broth and whole and semiskim milk was investigated using a laboratory-scale UV machine that incorporated static mixers within UV-penetrable pipes. UV treatment proved to be less effective in killing M. avium subsp. paratuberculosis suspended in milk (0.5- to 1.0-log10 reduction per 1,000 mJ/ml) than that suspended in Middlebrook 7H9 broth (2.5- to 3.3-log10 reduction per 1,000 mJ/ml). The FASTPlaqueTB phage assay provided more rapid enumeration of surviving M. avium subsp. paratuberculosis (within 24 h) than culture on Herrold's egg yolk medium (6 to 8 weeks). Despite the fact that plaque counts were consistently 1 to 2 log10 lower than colony counts throughout the study, UV inactivation rates for M. avium subsp. paratuberculosis derived using the phage assay and culture results were not significantly different (P = 0.077). PMID:17435001

  7. UV light inactivation of Mycobacterium avium subsp. paratuberculosis in milk as assessed by FASTPlaqueTB phage assay and culture.

    Science.gov (United States)

    Altic, Leslie C; Rowe, Michael T; Grant, Irene R

    2007-06-01

    UV light inactivation of Mycobacterium avium subsp. paratuberculosis in Middlebrook 7H9 broth and whole and semiskim milk was investigated using a laboratory-scale UV machine that incorporated static mixers within UV-penetrable pipes. UV treatment proved to be less effective in killing M. avium subsp. paratuberculosis suspended in milk (0.5- to 1.0-log(10) reduction per 1,000 mJ/ml) than that suspended in Middlebrook 7H9 broth (2.5- to 3.3-log(10) reduction per 1,000 mJ/ml). The FASTPlaqueTB phage assay provided more rapid enumeration of surviving M. avium subsp. paratuberculosis (within 24 h) than culture on Herrold's egg yolk medium (6 to 8 weeks). Despite the fact that plaque counts were consistently 1 to 2 log(10) lower than colony counts throughout the study, UV inactivation rates for M. avium subsp. paratuberculosis derived using the phage assay and culture results were not significantly different (P = 0.077).

  8. Persistence of Mycobacterium avium subsp. paratuberculosis in soil, crops, and ensiled feed following manure spreading on infected dairy farms.

    Science.gov (United States)

    Fecteau, Marie-Eve; Hovingh, Ernest; Whitlock, Robert H; Sweeney, Raymond W

    2013-11-01

    The goal of this study was to determine the persistence of Mycobacterium avium subsp. paratuberculosis (MAP) in soil, crops, and ensiled feeds following manure spreading. This bacterium was often found in soil samples, but less frequently in harvested feeds and silage. Spreading of manure on fields used for crop harvest is preferred to spreading on grazing pastures.

  9. Gamma-delta T cell responses in subclinical and clinical stages of Bovine Mycobacterium Avium Paratuberculosis infection

    Science.gov (United States)

    The early immune response to Mycobacterium avium subsp. paratuberculosis (MAP) in cattle is characterized by a Th1-like immune response effective in controlling bacterial proliferation during the subclinical stage of infection. In young calves nearly 60% of circulating lymphocytes are gamma delta T ...

  10. Immunization with a DNA Vaccine Cocktail Induces a Th1 Response and Protects Mice Against Mycobacterium avium subsp. paratuberculosis Challenge

    Science.gov (United States)

    Several novel antigens of Mycobacterium avium subsp. paratuberculosis have been studied as vaccine components and their immunogenicity has been evaluated. Previously, we reported that 85 antigen complex (85A, 85B, and 85C), superoxide dismutase (SOD), and 35kDa protein could induce significant lymph...

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

    Directory of Open Access Journals (Sweden)

    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.

  12. 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...... studies aiming at estimating the occurrence of MAP in on-farm BTM and IM by meta-analysis. A total of 692 articles were identified through electronic databases and initially screened using title and abstract. The quality of the 61 potentially relevant articles was assessed using full text and 31 articles...

  13. Protein Kinase G Induces an Immune Response in Cows Exposed to Mycobacterium avium Subsp. paratuberculosis

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    Horacio Bach

    2018-01-01

    Full Text Available To establish infection, pathogens secrete virulence factors, such as protein kinases and phosphatases, to modulate the signal transduction pathways used by host cells to initiate immune response. The protein MAP3893c is annotated in the genome sequence of Mycobacterium avium subspecies paratuberculosis (MAP, the causative agent of Johne’s disease, as the serine/threonine protein kinase G (PknG. In this work, we report that PknG is a functional kinase that is secreted within macrophages at early stages of infection. The antigen is able to induce an immune response from cattle exposed to MAP in the form of interferon gamma production after stimulation of whole blood with PknG. These findings suggest that PknG may contribute to the pathogenesis of MAP by phosphorylating macrophage signalling and/or adaptor molecules as observed with other pathogenic mycobacterial species.

  14. Apparent prevalence of beef carcasses contaminated with Mycobacterium avium subsp. paratuberculosis sampled from Danish slaughter cattle

    DEFF Research Database (Denmark)

    Okura, Hisako; Toft, Nils; Pozzato, Nicola

    2011-01-01

    Presence of Mycobacterium avium subsp. paratuberculosis (MAP) in beef has been reported as a public health concern because asymptomatically infected cattle may contain MAP in tissues that are used for human consumption. Associations between MAP carcasses contamination and animal characteristics...... such as age, breed, production type, and carcass classification were assessed. Cheek muscles from 501 carcasses were sampled cross-sectionally at a Danish abattoir and tested for presence of viable MAP and MAP DNA by bacterial culture and IS900 realtime PCR, respectively. Cheek muscle tissues from carcasses...... of two dairy cows were positive by culture whereas 4% of the animals were estimated with =10¿CFU/gram muscle based on realtime PCR. Age was found to be associated with carcass contamination with MAP. The observed viable MAP prevalence in beef carcasses was low. However, detection of MAP and MAP DNA...

  15. Apparent Prevalence of Beef Carcasses Contaminated with Mycobacterium avium subsp. paratuberculosis Sampled from Danish Slaughter Cattle

    Directory of Open Access Journals (Sweden)

    Hisako Okura

    2011-01-01

    Full Text Available 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 such as age, breed, production type, and carcass classification were assessed. Cheek muscles from 501 carcasses were sampled cross-sectionally at a Danish abattoir and tested for presence of viable MAP and MAP DNA by bacterial culture and IS900 realtime PCR, respectively. Cheek muscle tissues from carcasses of two dairy cows were positive by culture whereas 4% of the animals were estimated with ≥10 CFU/gram muscle based on realtime PCR. Age was found to be associated with carcass contamination with MAP. The observed viable MAP prevalence in beef carcasses was low. However, detection of MAP and MAP DNA in muscle tissues suggested that bacteremia occurred in slaughtered cattle.

  16. Gold nanoparticle-based probes for the colorimetric detection of Mycobacterium avium subspecies paratuberculosis DNA.

    Science.gov (United States)

    Ganareal, Thenor Aristotile Charles S; Balbin, Michelle M; Monserate, Juvy J; Salazar, Joel R; Mingala, Claro N

    2018-02-12

    Gold nanoparticle (AuNP) is considered to be the most stable metal nanoparticle having the ability to be functionalized with biomolecules. Recently, AuNP-based DNA detection methods captured the interest of researchers worldwide. Paratuberculosis or Johne's disease, a chronic gastroenteritis in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP), was found to have negative effect in the livestock industry. In this study, AuNP-based probes were evaluated for the specific and sensitive detection of MAP DNA. AuNP-based probe was produced by functionalization of AuNPs with thiol-modified oligonucleotide and was confirmed by Fourier-Transform Infrared (FTIR) spectroscopy. UV-Vis spectroscopy and Scanning Electron Microscopy (SEM) were used to characterize AuNPs. DNA detection was done by hybridization of 10 μL of DNA with 5 μL of probe at 63 °C for 10 min and addition of 3 μL salt solution. The method was specific to MAP with detection limit of 103 ng. UV-Vis and SEM showed dispersion and aggregation of the AuNPs for the positive and negative results, respectively, with no observed particle growth. This study therefore reports an AuNP-based probes which can be used for the specific and sensitive detection of MAP DNA. Copyright © 2018 Elsevier Inc. All rights reserved.

  17. Prevalence of Mycobacterium avium subspecies paratuberculosis and hepatitis E in New World camelids in Austria.

    Science.gov (United States)

    Stanitznig, A; Khol, J L; Lambacher, B; Franz, S; Wittek, T; Kralik, P; Slana, I; Vasickova, P

    2017-07-07

    Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of paratuberculosis in domestic ruminants and New World Camelids (NWC). Hepatitis E virus (HEV) is an important public health concern worldwide. The virus has been identified in several species, some of them serving as a reservoir for zoonotic HEV strains. Husbandry and breeding of llamas and alpacas have increased in Austria in recent years. Therefore, the aim of the present study was to evaluate the prevalence of MAP and HEV in NWC in Austria. Altogether 445 animals, originating from 78 farms were enrolled in the study. Of the animals sampled, 184 (41.35%) were llamas and 261 (58.65%) were alpacas. 443 blood samples for MAP-ELISA and 399 faecal samples for quantitative PCR (qPCR) and culture for MAP as well as for HEV detection by RT-qPCR have been collected. All of the 399 animals tested for shedding of MAP were negative by faecal solid culture. Using qPCR, 15 (3.8%) of the animals were MAP positive and 384 (96.2%) negative. Out of the 443 serum samples examined for specific antibodies against MAP by ELISA, 6 (1.4%) were positive, 1 (0.2%) was questionable and 436 (98.4%) samples were negative. All faecal samples were tested negative for HEV.

  18. Genetic variation and heritability of the antibody response to Mycobacterium avium subspecies paratuberculosis in Danish Hostein cows

    DEFF Research Database (Denmark)

    Mortensen, Hanne; Nielsen, Søren Saxmose; Berg, Peer

    2004-01-01

    The purpose of this study was to estimate the genetic variation and the heritability of the ability to establish an immune response by producing antibodies to Mycobacterium avium subsp. paratuberculosis. Antibody levels were determined using an ELISA and measuring optical density (OD) values from...... milk samples of 11,535 cows from 99 herds. The pedigree of the 11,535 cows and information about days in milk, parity, milk yield, and others were obtained from the Danish Cattle database. The statistical analyses were made using a bivariate mixed animal model. The bivariate model with daily milk yield...... and OD as dependent variables showed a significant heritability of the ability to produce Mycobacterium avium subsp. paratuberculosis antibodies of 0.102 (genetic variance = 0.054) and a nonsignificant genetic correlation of −0.037 between daily milk yield and OD. When a sire model was used...

  19. 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.

  20. New polymorphisms within the variable number tandem repeat (VNTR) 7 locus of Mycobacterium avium subsp. paratuberculosis.

    Science.gov (United States)

    Fawzy, Ahmad; Zschöck, Michael; Ewers, Christa; Eisenberg, Tobias

    2016-06-01

    Variable number tandem repeat (VNTR) is a frequently employed typing method of Mycobacterium avium paratuberculosis (MAP) isolates. Based on whole genome sequencing in a previous study, allelic diversity at some VNTR loci seems to over- or under-estimate the actual phylogenetic variance among isolates. Interestingly, two closely related isolates on one farm showed polymorphism at the VNTR 7 locus, raising concerns about the misleading role that it might play in genotyping. We aimed to investigate the underlying basis of VNTR 7-polymorphism by analyzing sequence data for published genomes and field isolates of MAP and other M. avium complex (MAC) members. In contrast to MAP strains from cattle, strains from sheep displayed an "imperfect" repeat within VNTR 7, which was identical to respective allele types in other MAC genomes. Subspecies- and strain-specific single nucleotide polymorphisms (SNPs) and two novel (16 and 56 bp) repeats were detected. Given the combination of the three existing repeats, there are at least five different patterns for VNTR 7. The present findings highlight a higher polymorphism and probable instability of VNTR 7 locus that needs to be considered and challenged in future studies. Until then, sequencing of this locus in future studies is important to correctly assign the underlying allele types.(1). Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Description of the Infection Status in a Norwegian Cattle Herd Naturally Infected by Mycobacterium avium subsp. paratuberculosis

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    Nyberg O

    2005-03-01

    Full Text Available The Norwegian surveillance and control programme for paratuberculosis revealed 8 seroreactors in a single dairy cattle herd that had no clinical signs of Mycobacterium avium subsp. paratuberculosis (M. a. paratuberculosis infection. Paratuberculosis had been a clinical problem in goats several years previously in this herd. All 45 cattle were culled and a thorough investigation of the infection status was conducted by the use of interferon-γ (IFN-γ immunoassay, measurement of antibodies, and pathological and bacteriological examination. In the IFN-γ immunoassay, 9 animals gave positive results, and 13 were weakly positive, while 19 animals were negative. In the serological test,10 animals showed positive reactions, and 5 were doubtful, while 30 animals gave negative reactions. There appeared to be a weak trend toward younger animals having raised IFN-γ and older animals having raised serological tests. Histopathological lesions compatible with paratuberculosis were diagnosed in 4 animals aged between 4 and 9 years. Three of these animals had positive serological reaction and one animal gave also positive results in the IFN-γ immunoassay. Infection was confirmed by isolation of M. a. paratuberculosis from 2 of these 4 animals. One single bacterial isolate examined by restriction fragment length polymorphism (RFLP had the same profile, B-C1, as a strain that had been isolated from a goat at the same farm several years previously. Despite many animals being positive in one or both of the immunological tests, indicative of a heavily infected herd, none of the animals showed clinical signs and only one cow was shown to be shedding bacteria. A cross-reaction with other mycobacteria might have caused some of the immunoreactions in these animals. It is also possible that the Norwegian red cattle breed is resistant to clinical infection with M. a. paratuberculosis.

  2. Long-term detection of Mycobacterium avium subspecies paratuberculosis in individual and bulk tank milk from a dairy herd with a low prevalence of Johne's disease.

    Science.gov (United States)

    Khol, J L; Wassertheurer, M; Sodoma, E; Revilla-Fernández, S; Damoser, J; Osterreicher, E; Dünser, M; Kleb, U; Baumgartner, W

    2013-06-01

    Mycobacterium avium ssp. paratuberculosis (MAP) causes Johne's disease (JD) in ruminants and is shed into the milk of infected cows, which contributes to the controversial discussion about a possible link between MAP and Crohn's disease in humans. The aim of the study was to investigate the risk for the entry of MAP in the food chain via milk from dairy farms with subclinical JD. Therefore, the occurrence of MAP in the milk of a dairy herd with a low prevalence of JD was studied in single and bulk tank milk samples over a period of 23 mo and compared with MAP shedding into feces. Milk, fecal, and blood samples were taken from all cows older than 1.5 yr of age at the beginning and the end of the trial and analyzed for MAP or specific antibodies. In addition, 63 cows (33 MAP infected and 30 MAP noninfected) were selected for monthly sampling. Raw and pasteurized bulk tank milk samples were collected on a monthly basis. The milk samples were tested for MAP by real-time quantitative PCR (qPCR), and the fecal samples were tested for bacterial shedding by qPCR or solid culture. Based on the results of the herd investigations, the prevalence of cows shedding MAP was around 5%; no cases of clinical JD were observed during the study period. The results of the ELISA showed high variation, with 2.1 to 5.1% positive milk samples and 14.9 to 18.8% ELISA-positive blood samples. Monthly milk sampling revealed low levels of MAP shedding into the individual milk samples of both MAP-infected and noninfected cows, with only 13 cows shedding the bacterium into milk during the study period. Mycobacterium avium ssp. paratuberculosis was not detected by qPCR in any raw or pasteurized bulk tank milk sample throughout the study. A significant positive association could be found between MAP shedding into milk and feces. From the results of the present study, it can be concluded that MAP is only shed via milk in a small proportion of cows with subclinical JD for a limited period of time and

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

    Science.gov (United States)

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

    2011-03-24

    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 isolates using PCR-based methods detecting genetic elements called Variable-Number Tandem Repeats (VNTRs) and Mycobacterial Interspersed Repetitive Units (MIRUs) to determine if multiple MAP strains can coexist on farms with endemic MAP infection. For 52 temporal isolates originating from infected cattle from 32 commercial dairy herds with known trading history, MIRU-VNTR analysis was applied at 10 loci of which six showed variation. Within the group of 52 isolates, 17 different MIRU-VNTR patterns were detected. One MIRU-VNTR pattern was found in 29 isolates, one pattern in four isolates, one pattern in three isolates, two times one MIRU-VNTR pattern was found occurring in two isolates, and 12 patterns were found only once. Eleven herds provided multiple isolates. In five herds a single MIRU-VNTR pattern was detected among multiple isolates whereas in six herds more than one pattern was found. This study confirms that between dairy farms as well as within dairy farms, infected animals shed MAP with different MIRU-VNTR patterns. Analysis of trading history and age within herds indicated that cows born within the same birth cohort can be infected with MAP strains exhibiting variations in the number of MIRU-VNTR repeats. These data indicate that such multiple genotypes of MAP can coexist within one herd. Copyright © 2010 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    Trefz, Phillip; Koehler, Heike; Klepik, Klaus; Moebius, Petra; Reinhold, Petra; Schubert, Jochen K; Miekisch, Wolfram

    2013-01-01

    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 diagnose MAP

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

    Directory of Open Access Journals (Sweden)

    Phillip Trefz

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

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

    OpenAIRE

    Ramírez G, René; Maldonado E, Juan

    2013-01-01

    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...

  7. 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.

  8. Causation of Crohn’s Disease by Mycobacterium avium Subspecies Paratuberculosis

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    John Hermon-Taylor

    2000-01-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (MAP is a member of the M avium complex (MAC. It differs genetically from other MAC in having 14 to 18 copies of IS900 and a single cassette of DNA involved in the biosynthesis of surface carbohydrate. Unlike other MAC, MAP is a specific cause of chronic inflammation of the intestine in many animal species, including primates. The disease ranges from pluribacillary to paucimicrobial, with chronic granulomatous inflammation like leprosy in humans. MAP infection can persist for years without causing clinical disease. The herd prevalence of MAP infection in Western Europe and North America is reported in the range 21% to 54%. These subclinically infected animals shed MAP in their milk and onto pastures. MAP is more robust than tuberculosis, and the risk that is conveyed to human populations in retail milk and in domestic water supplies is high. MAP is harboured in the ileocolonic mucosa of a proportion of normal people and can be detected in a high proportion of full thickness samples of inflamed Crohn’s disease gut by improved culture systems and IS900 polymerase chain reaction if the correct methods are used. MAP in Crohn’s disease is present in a protease-resistant nonbacillary form, can evade immune recognition and probably causes an immune dysregulation. As with other MAC, MAP is resistant to most standard antituberculous drugs. Treatment of Crohn’s disease with combinations of drugs more active against MAC such as rifabutin and clarithromycin can bring about a profound improvement and, in a few cases, apparent disease eradication. New drugs as well as effective MAP vaccines for animals and humans are needed. The problems caused by MAP constitute a public health issue of tragic proportions for which a range of remedial measures are urgently needed.

  9. Fabrication of a Novel Conductometric Biosensor for Detecting Mycobacterium avium subsp. paratuberculosis Antibodies

    Directory of Open Access Journals (Sweden)

    Steven Bolin

    2008-09-01

    Full Text Available Johne’s disease (JD is one of the most costly bacterial diseases in cattle. In the U.S., economic losses from the disease have been estimated to exceed $1,500,000,000 per year, mainly from the effects of reduced milk production. Current diagnostic tests for JD are laboratory based and many of those tests require specialized equipment and training. Development of rapid and inexpensive diagnostic assays, which are adapted for point-ofcare applications, would aid in the control of JD. In this study, a polyaniline (Pani-based conductometric biosensor, in an immunomigration format, was fabricated for the detection of serum antibody (IgG against the causal organism of JD, Mycobacterium avium subsp. paratuberculosis (MAP. Immobilized Mycobacterium avium purified proteins in the capture membrane were used to detect MAP IgG, previously bound with Pani/anti-bovine IgG* conjugate in the conjugate membrane. After detection, the Pani in the sandwiched captured complex bridges an electrical circuit between the silver electrodes, flanking the capture membrane. The electrical conductance, caused by Pani, was measured as drop in electrical resistance. Testing of the biosensor with known JD positive and negative serum samples demonstrated a significant difference in the mean resistance observed between the groups. This proof-of-concept study demonstrated that a conductometric biosensor could detect MAP IgG in 2 minutes. The biosensor’s speed of detection and the equipment involved would, among other things, support its application towards the various point-ofcare opportunities aimed at JD management and control.

  10. Detection of Mycobacterium avium subsp. paratuberculosis in bovine milk from the state of Pernambuco, Brazil.

    Science.gov (United States)

    Albuquerque, Pedro Paulo Feitosa de; Santos, André de Souza; Souza Neto, Orestes Luiz de; Kim, Pomy de Cássia Peixoto; Cavalcanti, Erika Fernanda Torres Samico Fernandes; Oliveira, Júnior Mário Baltazar de; Mota, Rinaldo Aparecido; Júnior, José Wilton Pinheiro

    The aim of this study was to detect the IS900 region of Mycobacterium avium subsp. paratuberculosis (MAP) in bovine milk samples using real-time polymerase chain reaction (qPCR) and conventional PCR, and to study the agreement between these tests. A total of 121 bovine milk samples were collected from herds considered positive for MAP, from the State of Pernambuco, Brazil. MAP DNA was detected in 20 samples (16.5%) using conventional PCR and in 34 samples (28.1%) using qPCR. MAP DNA was detected in all of the 6 animal farms studied. Moderate agreement was found between qPCR and conventional PCR results, where the sensitivity and specificity of conventional PCR in relation to qPCR were 50% and 96.6%, respectively. Thus, the IS900 region of MAP was found in bovine milk samples from the State of Pernambuco. To the best of our knowledge, this is the first report of MAP DNA found in bovine milk in Northeast Brazil. We also demonstrated the qPCR technique is more sensitive than conventional PCR with respect to detection of MAP in milk samples. Copyright © 2016. Published by Elsevier Editora Ltda.

  11. Leaching of Mycobacterium avium subsp. paratuberculosis in Soil under In Vitro Conditions

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    Eran A. Raizman

    2011-01-01

    Full Text Available Mycobacterium avium subsp paratuberculosis (Map, the causative agent of Johne's disease, has a robust ability to survive in the environment. However, the ability of Map to migrate through soil to drainage tiles or ground water, leave the farm, and leak into local watersheds is inadequately documented. In order to assess the ability of Map to leach through soil, two laboratory experiments were conducted. In the first study, 8 columns (30 cm long each of a sandy loam soil were treated with pure cultures of Map. Two soil moisture levels and two Map concentrations were used. The columns were leached with 500 mL of water once a week for three weeks, the leachate was collected, and detection analysis was conducted. In the second experiment, manure from Map negative cows (control and Map high shedder cows (treatment were deposited on 8 similar columns and the columns were leached with 500 mL of water once a week for four weeks. Map detection and numeration in leachate samples were done with RT-PCR and culture techniques, respectively. Using RT-PCR, Map could be detected in the leachates in both experiments for several weeks but could only be recovered using culture techniques in experiment one. Combined, these experiments indicate the potential for Map to move through soil as a result of rainfall or irrigation following application.

  12. Leaching of Mycobacterium avium Subsp paratuberculosis in Soil under In Vitro Conditions.

    Science.gov (United States)

    Raizman, Eran A; Habteselassie, Mussie Y; Wu, Ching C; Lin, Tsang L; Negron, M; Turco, Ronald F

    2011-01-01

    Mycobacterium avium subsp paratuberculosis (Map), the causative agent of Johne's disease, has a robust ability to survive in the environment. However, the ability of Map to migrate through soil to drainage tiles or ground water, leave the farm, and leak into local watersheds is inadequately documented. In order to assess the ability of Map to leach through soil, two laboratory experiments were conducted. In the first study, 8 columns (30 cm long each) of a sandy loam soil were treated with pure cultures of Map. Two soil moisture levels and two Map concentrations were used. The columns were leached with 500 mL of water once a week for three weeks, the leachate was collected, and detection analysis was conducted. In the second experiment, manure from Map negative cows (control) and Map high shedder cows (treatment) were deposited on 8 similar columns and the columns were leached with 500 mL of water once a week for four weeks. Map detection and numeration in leachate samples were done with RT-PCR and culture techniques, respectively. Using RT-PCR, Map could be detected in the leachates in both experiments for several weeks but could only be recovered using culture techniques in experiment one. Combined, these experiments indicate the potential for Map to move through soil as a result of rainfall or irrigation following application.

  13. Mycobacterium avium subsp. paratuberculosis in an Italian Cohort of Type 1 Diabetes Pediatric Patients

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    Maria Luisa Manca Bitti

    2012-01-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (MAP is the etiological agent of Johne’s disease in ruminants. Recent studies have linked MAP to type 1 diabetes (T1D in the Sardinian population. The aim of this study was to investigate the prevalence of MAP infection in a T1D cohort from continental Italy compared with healthy control subjects. 247 T1D subjects and 110 healthy controls were tested for the presence of MAP. MAP DNA was detected using IS900-specific polymerase chain reaction (PCR. The presence of antibodies towards a MAP antigen, heparin binding hemoagglutinin (HBHA, was detected by ELISA. We demonstrated a higher MAP DNA prevalence in plasma samples from T1D patients and a stronger immune response towards MAP HBHA, compared with healthy control subjects. Moreover, in the recent onset patients, we observed an association between anti-MAP antibodies and HLA DQ2 (DQA1 0201/DQB1 0202. These findings taken together support the hypothesis of MAP as an environmental risk factor for the development of T1D in genetically predisposed subjects, probably involving a mechanism of molecular mimicry between MAP antigens and pancreatic islet β-cells.

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

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    Christopher D Johnston

    2014-09-01

    Full Text Available It is well documented that open reading frames containing high GC content show poor expression in A+T rich hosts. Specifically, G+C-rich codon usage is a limiting factor in heterologous expression of Mycobacterium avium subsp. paratuberculosis (MAP proteins using Lactobacillus salivarius. However, re-engineering opening reading frames through synonymous substitutions can offset codon bias and greatly enhance MAP protein production in this host. In this report, we demonstrate that codon-usage manipulation of two MAP genes (MAP2121c and MAP3733c can enhance the heterologous expression of two antigens (MMP and MptD respectively, analogous to the form to which they are produced natively by MAP bacilli. When heterologously over-expressed, antigenic determinants were preserved in synthetic MMP proteins as shown by monoclonal antibody mediated ELISA. Moreover, MMP is a membrane protein in MAP, which is also targeted to the cellular surface of recombinant L. salivarius at levels comparable to MAP. Additionally, codon optimised MptD displayed the tendency to associate with the cytoplasmic membrane boundary under confocal microscopy and the intracellularly accumulated protein selectively adhered with the MptD-specific bacteriophage fMptD. This work demonstrates there is potential for L. salivarius as a viable antigen delivery vehicle for MAP, which may provide an effective mucosal vaccine against Johne’s disease.

  15. Contamination of food products with Mycobacterium avium paratuberculosis: a systematic review.

    Science.gov (United States)

    Eltholth, M M; Marsh, V R; Van Winden, S; Guitian, F J

    2009-10-01

    Although a causal link between Mycobacterium avium subspecies paratuberculosis (MAP) and Crohn's disease has not been proved, previous studies suggest that the potential routes of human exposure to MAP should be investigated. We conducted a systematic review of literature concerning the likelihood of contamination of food products with MAP and the likely changes in the quantity of MAP in dairy and meat products along their respective production chains. Relevant data were extracted from 65 research papers and synthesized qualitatively. Although estimates of the prevalence of Johne's disease are scarce, particularly for non-dairy herds, the available data suggest that the likelihood of contamination of raw milk with MAP in most studied regions is substantial. The presence of MAP in raw and pasteurized milk has been the subject of several studies which show that pasteurized milk is not always MAP-free and that the effectiveness of pasteurization in inactivating MAP depends on the initial concentration of the agent in raw milk. The most recent studies indicated that beef can be contaminated with MAP via dissemination of the pathogen in the tissues of infected animals. Currently available data suggests that the likelihood of dairy and meat products being contaminated with MAP on retail sale should not be ignored.

  16. Inferring biomarkers for Mycobacterium avium subsp. paratuberculosis infection and disease progression in cattle using experimental data

    Science.gov (United States)

    Magombedze, Gesham; Shiri, Tinevimbo; Eda, Shigetoshi; Stabel, Judy R.

    2017-03-01

    Available diagnostic assays for Mycobacterium avium subsp. paratuberculosis (MAP) have poor sensitivities and cannot detect early stages of infection, therefore, there is need to find new diagnostic markers for early infection detection and disease stages. We analyzed longitudinal IFN-γ, ELISA-antibody and fecal shedding experimental sensitivity scores for MAP infection detection and disease progression. We used both statistical methods and dynamic mathematical models to (i) evaluate the empirical assays (ii) infer and explain biological mechanisms that affect the time evolution of the biomarkers, and (iii) predict disease stages of 57 animals that were naturally infected with MAP. This analysis confirms that the fecal test is the best marker for disease progression and illustrates that Th1/Th2 (IFN-γ/ELISA antibodies) assays are important for infection detection, but cannot reliably predict persistent infections. Our results show that the theoretical simulated macrophage-based assay is a potential good diagnostic marker for MAP persistent infections and predictor of disease specific stages. We therefore recommend specifically designed experiments to test the use of a based assay in the diagnosis of MAP infections.

  17. Mycobacterium avium Subspecies paratuberculosis: Human Exposure through Environmental and Domestic Aerosols

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    Glenn Rhodes

    2014-07-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (Map causes Johne’s disease in animals and is significantly associated with Crohn’s disease (CD in humans. Our previous studies have shown Map to be present in U.K. rivers due to land deposition from chronic livestock infection and runoff driven by rainfall. The epidemiology of CD in Cardiff showed a significant association with the River Taff, in which Map can be detected on a regular basis. We have previously hypothesized that aerosols from the river might influence the epidemiology of CD. In this preliminary study, we detected Map by quantitative PCR in one of five aerosol samples collected above the River Taff. In addition, we examined domestic showers from different regions in the U.K. and detected Map in three out of 30 independent samples. In detecting Map in river aerosols and those from domestic showers, this is the first study to provide evidence that aerosols are an exposure route for Map to humans and may play a role in the epidemiology of CD.

  18. Comparison of rapid diagnostic tests to detect Mycobacterium avium subsp. paratuberculosis disseminated infection in bovine liver.

    Science.gov (United States)

    Zarei, Mehdi; Ghorbanpour, Masoud; Tajbakhsh, Samaneh; Mosavari, Nader

    2017-08-01

    Mycobacterium avium subsp. paratuberculosis (MAP) causes Johne's disease, a chronic enteritis in cattle and other domestic and wild ruminants. The presence of MAP in tissues other than intestines and associated lymph nodes, such as meat and liver, is a potential public health concern. In the present study, the relationship between the results of rapid diagnostic tests of the Johne's disease, such as serum ELISA, rectal scraping PCR, and acid-fast staining, and the presence of MAP in liver was evaluated. Blood, liver, and rectal scraping samples were collected from 200 slaughtered cattle with unknown Johne's disease status. ELISA was performed to determine the MAP antibody activity in the serum. Acid-fast staining was performed on rectal scraping samples, and PCR was performed on rectal scraping and liver samples. PCR-positive liver samples were used for mycobacterial culture. Overall, the results of this study demonstrated that MAP can be detected and cultured from liver of slaughtered cattle and rapid diagnostic tests of Johne's disease have limited value in detecting cattle with MAP infection in liver. These findings show that the presence of MAP in liver tissue may occur in cows with negative results for rapid diagnostic tests and vice versa. Hence, liver might represent another possible risk of human exposure to MAP. Given concerns about a potential zoonotic role for MAP, these results show the necessity to find new methods for detecting cattle with MAP disseminated infection.

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

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    Arsenault, Ryan J; Maattanen, Pekka; Daigle, Joanna; Potter, Andrew; Griebel, Philip; Napper, Scott

    2014-05-15

    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 to control JD through traditional animal management practices are complicated by MAP's ability to cause long-term environmental contamination as well as difficulties associated with diagnosis of JD in the pre-clinical stages. As such, there is particular emphasis on the development of an effective vaccine. This is a daunting challenge, in large part due to MAP's ability to subvert protective host immune responses. Accordingly, there is a priority to understand MAP's interaction with the bovine host: this may inform rational targets and approaches for therapeutic intervention. Here we review the early host defenses encountered by MAP and the strategies employed by the pathogen to avert or subvert these responses, during the critical period between ingestion and the establishment of persistent infection in macrophages.

  20. Mycobacterium avium subesp. paratuberculosis: uma preocupação para a indústria de laticínios

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    Márcio Ferraz Cunha

    2009-04-01

    Full Text Available Mycobacterium avium subesp. paratuberculosis é conhecida como o agente etiológico da doença de Johne, ou paratuberculose, que afeta principalmente animais ruminantes. É integrante da família Mycobacteriaceae, da qual também fazem parte a M. tuberculosis e a M. bovis, responsáveis pela tuberculose humana e bovina, respectivamente. Foi sugerido que a M. paratuberculosis poderia estar envolvida na patogênese da doença de Crohn, a qual possui sintomas similares à paratuberculose, mas afeta seres humanos. Como o microrganismo pode ser excretado no leite de animais infectados, o primeiro passo foi avaliar a sua termoresistência. Alguns estudos indicaram que a bactéria sobrevive ao tratamento térmico da pasteurização HTST (72ºC/15 s. Entretanto, os estudos existentes na literatura científica até o momento não permitem afirmar que M. paratuberculosis seja responsável pela doença de Crohn, bem como apresentam dúvidas sobre a termoresistência dessa bactéria. A realização de mais pesquisas sobre este microrganismo é de fundamental importância, com o objetivo de orientar a produção de produtos lácteos isentos de contaminação por M. paratuberculosis.

  1. Development and Evaluation of a Novel Multicopy-Element-Targeting Triplex PCR for Detection of Mycobacterium avium subsp. paratuberculosis in Feces

    Science.gov (United States)

    Garrido, Joseba M.; Molina, Elena; Geijo, María V.; Elguezabal, Natalia; Vázquez, Patricia; Juste, Ramón A.

    2014-01-01

    The enteropathy called paratuberculosis (PTB), which mainly affects ruminants and has a worldwide distribution, is caused by Mycobacterium avium subsp. paratuberculosis. This disease significantly reduces the cost-effectiveness of ruminant farms, and therefore, reliable and rapid detection methods are needed to control the spread of the bacterium in livestock and in the environment. The aim of this study was to identify a specific and sensitive combination of DNA extraction and amplification to detect M. avium subsp. paratuberculosis in feces. Negative bovine fecal samples were inoculated with increasing concentrations of two different bacterial strains (field and reference) to compare the performance of four extraction and five amplification protocols. The best results were obtained using the JohnePrep and MagMax extraction kits combined with an in-house triplex real-time PCR designed to detect IS900, ISMap02 (an insertion sequence of M. avium subsp. paratuberculosis present in 6 copies per genome), and an internal amplification control DNA simultaneously. These combinations detected 10 M. avium subsp. paratuberculosis cells/g of spiked feces. The triplex PCR detected 1 fg of genomic DNA extracted from the reference strain K10. The performance of the robotized version of the MagMax extraction kit combined with the IS900 and ISMap02 PCR was further evaluated using 615 archival fecal samples from the first sampling of nine Friesian cattle herds included in a PTB control program and followed up for at least 4 years. The analysis of the results obtained in this survey demonstrated that the diagnostic method was highly specific and sensitive for the detection of M. avium subsp. paratuberculosis in fecal samples from cattle and a very valuable tool to be used in PTB control programs. PMID:24727272

  2. 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...

  3. Predicting the role of IL-10 in the regulation of the adaptive immune responses in Mycobacterium avium subsp. paratuberculosis infections using mathematical models

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is an intracellular bacterial pathogen that causes Johne’s disease in cattle and other animals. Infection follows ingestion of the bacteria primarily through the fecal oral route and results in the colonization of the intestine and a granulomatous en...

  4. Development and validation of a liquid medium (M7H9C) for routine culture of Mycobacterium avium subsp. paratuberculosis to replace modified Bactec 12B medium.

    Science.gov (United States)

    Whittington, Richard J; Whittington, Ann-Michele; Waldron, Anna; Begg, Douglas J; de Silva, Kumi; Purdie, Auriol C; Plain, Karren M

    2013-12-01

    Liquid culture of Mycobacterium avium subsp. paratuberculosis from clinical samples, such as feces, is the most sensitive antemortem test for the diagnosis of Johne's disease in ruminants. In Australia, New Zealand, the United States, and some other countries, the Bactec 460 system with modified Bactec 12B medium (Becton, Dickinson) has been the most commonly used liquid culture system, but it was discontinued in 2012. In this study, a new liquid culture medium, M7H9C, was developed. It consists of a Middlebrook 7H9 medium base with added Casitone, albumin, dextrose, catalase, egg yolk, mycobactin J, and a cocktail of antibiotics. We found that polyoxyethylene stearate (POES) was not essential for the cultivation of M. avium subsp. paratuberculosis in either the Bactec 12B or the M7H9C medium. The limit of detection determined using pure cultures of the C and S strains of M. avium subsp. paratuberculosis was 7 bacilli per 50 μl inoculum in the two media. The new medium was validated using 784 fecal and tissue samples from sheep and cattle, >25% of which contained viable M. avium subsp. paratuberculosis. Discrepant results for the clinical samples between the two media were mostly associated with samples that contained medium was less than half the cost of the Bactec 12B medium and did not require regular examination during incubation, but a confirmatory IS900 PCR test had to be performed on every culture after the predetermined incubation period.

  5. 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

  6. 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.

  7. A rhodanine agent active against non-replicating intracellular Mycobacterium avium subspecies paratuberculosis

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    Bull Tim J

    2009-12-01

    Full Text Available Abstract Background Antibiotic therapy targeting chronic mycobacterial disease is often ineffective due to problems with the emergence of drug resistance and non-replicating persistent intracellular antibiotic resistant phenotypes. Strategies which include agents able to enhance host cell killing mechanisms could represent an alternative to conventional methods with the potential for host clearance if active against dormant phenotypes. Investigations of agents with potential activity against non-replicating mycobacteria however are restricted due to a need for assays that can assess bacterial viability without having to culture. Results This study describes the development and use of a pre16S ribosomal gene RNA/DNA ratio viability assay which is independent of the need for culture, supported by a novel thin layer accelerated mycobacterial colony forming method for determining viability and culturability of MAP in intracellular environments. We describe the use of these tools to demonstrate intracellular killing activity of a novel rhodanine agent (D157070 against the intracellular pathogen Mycobacterium avium subspecies paratuberculosis (MAP and show that the culturability of MAP decreases relative to its viability on intracellular entry suggesting the induction of a non-culturable phenotype. We further demonstrate that D157070, although having no direct activity against the culturability of extracellular MAP, can bind to cultured MAP cells and has significant influence on the MAP transcriptome, particularly with respect of δL associated genes. D157070 is shown to be taken up by bovine and human cells and able to enhance host cell killing, as measured by significant decreases in both culturability and viability of intracellular MAP. Conclusions This work suggests that pre16srRNA gene ratios represent a viable method for studying MAP viability. In addition, the rhodanine agent D157070 tested is non-toxic and enhances cell killing activity

  8. Dysbiosis of the Fecal Microbiota in Cattle Infected with Mycobacterium avium subsp. paratuberculosis.

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    Marie-Eve Fecteau

    Full Text Available Johne's disease (JD is a chronic, intestinal infection of cattle, caused by Mycobacterium avium subsp. paratuberculosis (MAP. It results in granulomatous inflammation of the intestinal lining, leading to malabsorption, diarrhea, and weight loss. Crohn's disease (CD, a chronic, inflammatory gastrointestinal disease of humans, has many clinical and pathologic similarities to JD. Dysbiosis of the enteric microbiota has been demonstrated in CD patients. It is speculated that this dysbiosis may contribute to the intestinal inflammation observed in those patients. The purpose of this study was to investigate the diversity patterns of fecal bacterial populations in cattle infected with MAP, compared to those of uninfected control cattle, using phylogenomic analysis. Fecal samples were selected to include samples from 20 MAP-positive cows; 25 MAP-negative herdmates; and 25 MAP-negative cows from a MAP-free herd. The genomic DNA was extracted; PCR amplified sequenced on a 454 Roche platform, and analyzed using QIIME. Approximately 199,077 reads were analyzed from 70 bacterial communities (average of 2,843 reads/sample. The composition of bacterial communities differed between the 3 treatment groups (P < 0.001; Permanova test. Taxonomic assignment of the operational taxonomic units (OTUs identified 17 bacterial phyla across all samples. Bacteroidetes and Firmicutes constituted more than 95% of the bacterial population in the negative and exposed groups. In the positive group, lineages of Actinobacteria and Proteobacteria increased and those of Bacteroidetes and Firmicutes decreased (P < 0.001. Actinobacteria was highly abundant (30% of the total bacteria in the positive group compared to exposed and negative groups (0.1-0.2%. Notably, the genus Arthrobacter was found to predominate Actinobacteria in the positive group. This study indicates that MAP-infected cattle have a different composition of their fecal microbiota than MAP-negative cattle.

  9. 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.

  10. Low genetic diversity of bovine Mycobacterium avium subspecies paratuberculosis isolates detected by MIRU-VNTR genotyping.

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    de Kruijf, Marcel; Lesniak, Olga N; Yearsley, Dermot; Ramovic, Elvira; Coffey, Aidan; O'Mahony, Jim

    2017-05-01

    Mycobacterial interspersed repetitive unit and variable number tandem repeat (MIRU-VNTR) has been developed as a simple, rapid and cost efficient molecular typing method to differentiate Mycobacterium avium subspecies paratuberculosis (MAP) isolates. The aim of this study was to determine the genomic diversity of MAP across the Republic of Ireland by utilising the MIRU-VNTR typing method on a large collection of MAP isolates. A total of 114 MAP isolates originated from 53 herds across 19 counties in the Republic of Ireland were genotyped based on eight established MIRU-VNTR loci. Four INMV groups were observed during this study. INMV 1 was found in 67 MAP isolates (58.8%) and INMV 2 was observed in 45 isolates (39.4%). INMV 3 and INMV 116 recorded only one isolate each (0.9%). The unique INMV 116 group has never been reported among herds thus far and the molecular pattern of the MAP isolate classified in INMV 116 showed a difference at the MIRU-VNTR X3 locus compared to the other three INMV groups observed. INMV 1, INMV 2 and INMV 3 are observed frequently in Europe and comprised 99.1% of the total MAP isolates characterised in this study, indicating that MAP exhibited low level of genetic diversity across the Republic of Ireland using the MIRU-VNTR method. By the implementation of SNP analysis or MLSSR as an additional typing method, MAP genetic diversity would increase. INMV 3 is unique to Ireland and whereas INMV 116 has never been previously reported among herds by MIRU-VNTR typing. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Intestinal infection following aerosol challenge of calves with Mycobacterium avium subspecies paratuberculosis

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    Eisenberg Susanne WF

    2011-12-01

    Full Text Available Abstract A challenge experiment was performed to investigate whether administration of Mycobacterium avium subsp. paratuberculosis (MAP via the respiratory route leads to MAP infection in calves. Eighteen calves from test negative dams were randomly allocated to four groups. Six calves were challenged with MAP nasally and six calves were challenged by transtracheal injection; three orally challenged calves served as positive controls, and three non challenged calves as negative controls. The challenge was performed as a nine-fold trickle dose, 107 CFU in total. Blood and faecal samples were collected frequently. Calves were euthanized three months post-challenge and extensively sampled. Blood samples were tested for the presence of antibodies and interferon gamma producing cells by ELISA. Faecal and tissue samples were cultured in a liquid culture system and the presence of MAP was confirmed by IS900 realtime PCR. Fourteen out of fifteen calves had no MAP antibody response. The negative controls remained negative; all positive controls became infected. Two nasally challenged calves showed a Purified Protein Derivative Avian (PPDA specific interferon gamma response. In all nasally challenged calves, MAP positive intestinal samples were detected. In three calves of the nasal group MAP positive retropharyngeal lymph nodes or tonsils were detected. In all calves of the transtracheal group MAP positive intestinal tissues were detected as well and three had a MAP positive tracheobronchial lymph node. These findings indicate that inhalation of MAP aerosols can result in infection. These experimental results may be relevant for transmission under field conditions since viable MAP has been detected in dust on commercial dairy farms.

  12. Transcriptional Profiling of Ileocecal Valve of Holstein Dairy Cows Infected with Mycobacterium avium subsp. Paratuberculosis.

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    Randy J Hempel

    Full Text Available Johne's disease is a chronic infection of the small intestine caused by Mycobacterium avium subspecies paratuberculosis (MAP, an intracellular bacterium. The events of pathogen survival within the host cell(s, chronic inflammation and the progression from asymptomatic subclinical stage to an advanced clinical stage of infection, are poorly understood. This study examines gene expression in the ileocecal valve (ICV of Holstein dairy cows at different stages of MAP infection. The ICV is known to be a primary site of MAP colonization and provides an ideal location to identify genes that are relevant to the progression of this disease. RNA was prepared from ICV tissues and RNA-Seq was used to compare gene transcription between clinical, subclinical, and uninfected control animals. Interpretation of the gene expression data was performed using pathway analysis and gene ontology categories containing multiple differentially expressed genes. Results demonstrated that many of the pathways that had strong differential gene expression between uninfected control and clinical cows were related to the immune system, such as the T- and B-cell receptor signaling, apoptosis, NOD-like receptor signaling, and leukocyte transendothelial migration pathways. In contrast, the comparison of gene transcription between control and subclinical cows identified pathways that were primarily involved in metabolism. The results from the comparison between clinical and subclinical animals indicate recruitment of neutrophils, up regulation of lysosomal peptidases, increase in immune cell transendothelial migration, and modifications of the extracelluar matrix. This study provides important insight into how cattle respond to a natural MAP infection at the gene transcription level within a key target tissue for infection.

  13. 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.

  14. A Mycobacterium avium subsp. paratuberculosis predicted serine protease is associated with acid stress and intraphagosomal survival

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    Abirami Kugadas

    2016-08-01

    Full Text Available AbstractThe ability to maintain intra-cellular pH is crucial for bacteria and other microbes to survive in diverse environments, particularly those that undergo fluctuations in pH. Mechanisms of acid resistance remain poorly understood in mycobacteria. Although studies investigating acid stress in M. tuberculosis are gaining traction, few center on Mycobacterium avium subsp. paratuberculosis (MAP, the etiological agent of chronic enteritis in ruminants. We identified a MAP acid stress response network involved in macrophage infection. The central node of this network was MAP0403, a predicted serine protease that shared an 86% amino acid identity with MarP in M. tuberculosis. Previous studies confirmed MarP as a serine protease integral to maintaining intra-bacterial pH and survival in acid in vitro and in vivo. We show that MAP0403 is upregulated in infected macrophage and MAC-T cells and coincided with phagosome acidification. Treatment of mammalian cells with bafilomcyin A1, a potent inhibitor of phagosomal vATPases, diminished MAP0403 transcription. MAP0403 expression was also noted in acidic medium. A surrogate host, M. smegmatis mc2 155, was designed to express MAP0403 and when exposed to either macrophages or in vitro acid stress had increase bacterial cell viability, which corresponds to maintenance of intra-bacterial pH in acidic (pH = 5 conditions. These data suggest that MAP0403 may be the equivalent of MarP in MAP. Future studies confirming MAP0403 as a serine protease and exploring its structure and possible substrates are warranted.

  15. 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.

  16. Evolution of the bovine TLR gene family and member associations with Mycobacterium avium subspecies paratuberculosis infection.

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    Colleen A Fisher

    Full Text Available Members of the Toll-like receptor (TLR gene family occupy key roles in the mammalian innate immune system by functioning as sentries for the detection of invading pathogens, thereafter provoking host innate immune responses. We utilized a custom next-generation sequencing approach and allele-specific genotyping assays to detect and validate 280 biallelic variants across all 10 bovine TLR genes, including 71 nonsynonymous single nucleotide polymorphisms (SNPs and one putative nonsense SNP. Bayesian haplotype reconstructions and median joining networks revealed haplotype sharing between Bos taurus taurus and Bos taurus indicus breeds at every locus, and specialized beef and dairy breeds could not be differentiated despite an average polymorphism density of 1 marker/158 bp. Collectively, 160 tagSNPs and two tag insertion-deletion mutations (indels were sufficient to predict 100% of the variation at 280 variable sites for both Bos subspecies and their hybrids, whereas 118 tagSNPs and 1 tagIndel predictively captured 100% of the variation at 235 variable sites for B. t. taurus. Polyphen and SIFT analyses of amino acid (AA replacements encoded by bovine TLR SNPs indicated that up to 32% of the AA substitutions were expected to impact protein function. Classical and newly developed tests of diversity provide strong support for balancing selection operating on TLR3 and TLR8, and purifying selection acting on TLR10. An investigation of the persistence and continuity of linkage disequilibrium (r2≥0.50 between adjacent variable sites also supported the presence of selection acting on TLR3 and TLR8. A case-control study employing validated variants from bovine TLR genes recognizing bacterial ligands revealed six SNPs potentially eliciting small effects on susceptibility to Mycobacterium avium spp paratuberculosis infection in dairy cattle. The results of this study will broadly impact domestic cattle research by providing the necessary foundation to

  17. Lactase persistence, NOD2 status and Mycobacterium avium subsp. paratuberculosis infection associations to Inflammatory Bowel Disease

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    Elguezabal Natalia

    2012-06-01

    Full Text Available Abstract Background Inflammatory Bowel Disease (IBD, which includes both Crohn’s disease (CD and ulcerative colitis (UC, is caused by a complex interplay involving genetic predisposition, environmental factors and an infectious agent. Mycobacterium avium subsp. paratuberculosis (MAP is a promising pathogen candidate since it produces a chronic intestinal inflammatory disease in ruminants that resembles CD in humans. MAP is a ubiquitous microorganism, although its presence in the food chain, especially in milk from infected animals, is what made us think that there could be an association between lactase persistence (LP and IBD. The LCT mutation has brought adaptation to dairy farming which in turn would have increased exposure of the population to infection by MAP. NOD2 gene mutations are highly associated to CD. Methods In our study, CD and UC patients and controls from the North of Spain were genotyped for the lactase gene (LCT and for three NOD-2 variants, R702W, G908R and Cins1007fs. MAP PCR was carried out in order to assess MAP infection status and these results were correlated with LCT and NOD2 genotypes. Results As for LP, no association was found with IBD, although UC patients were less likely to present the T/T−13910 variant compared to controls, showing a higher C-allele frequency and a tendency to lactase non-persistence (LNP. NOD2 mutations were associated to CD being the per-allele risk higher for the Cins1007fs variant. MAP infection was more extended among the healthy controls (45.2% compared to CD patients (21.38% and UC patients (19.04% and this was attributed to therapy. The Asturian CD cohort presented higher levels of MAP prevalence (38.6% compared to the Basque CD cohort (15.5%, differences also attributed to therapy. No interaction was found between MAP infection and LCT or NOD2 status. Conclusions We conclude that LP is not significantly associated with IBD, but that MAP infection and NOD2 do show not mutually

  18. 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

    Denmark is free of M. bovis, two of the vaccinated animals responded with higher IFN-γ levels when cultured with PPDb compared to PPDa. In conclusion, immunization with whole-cell MAP vaccines elicits both humoral and cell-mediated immune reactions, which may interfere with surveillance and diagnosis......Vaccination of cattle against Mycobacterium avium subsp. paratuberculosis (MAP) provides partial protection by delayed shedding of MAP and reduced numbers of clinically affected animals. The duration of vaccine induced immune response is not known. The primary objective of this study was therefore...... to characterize the long-term effect of whole-cell based vaccination against MAP on the immune response. A secondary objective was to evaluate whether immunodiagnosis of MAP and Mycobacterium bovis infections is affected by MAP vaccination.Two studies were performed: (1) A retrospective longitudinal study...

  19. 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

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

  20. 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.; Bull, Tim J.; 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...

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

    OpenAIRE

    Aida Chalesh (MSc); Keyvan Tadayon ( PhD

    2016-01-01

    Background and Objective: Paratuberculosis has been repeatedly reported from Iranian ruminant herds. The extrem fastidious nature of Mycobacterium avium subspecies paratuberculsos hinders genomic diversity studies of the pathogen. Short Sequence Repeat analysis is one of the genome-based approches recently developed to overcome this difficulty. In this study we describe the application of SSR genotyping on three Iranian MAP type strains plus the III & V vaccinal strain. Methods: A...

  2. Rapid identification of Mycobacterium avium ssp paratuberculosis laboratory strains by IS900-Nested polymerase chain reaction

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    Mohammad Mohammad Taheri

    2016-01-01

    Conclution: However, no amplification was observed with other strains. Two main achievements of this work are the development of an efficient means of differentiation between the six Razi laboratory mycobacterial strains and characterization of the genomic profile of these strains, a capability that is vital when cross contamination is potentially an important concern.

  3. Current status of Mycobacterium avium subspecies paratuberculosis infection in animals & humans in India: What needs to be done?

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    Ajay Vir Singh

    2016-01-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (MAP has emerged as a major health problem for domestic livestock and human beings. Reduced per animal productivity of domestic livestock seriously impacts the economics of dairy farming globally. High to very high bioload of MAP in domestic livestock and also in the human population has been reported from north India. Presence of live MAP bacilli in commercial supplies of raw and pasteurized milk and milk products indicates its public health significance. MAP is not inactivated during pasteurization, therefore, entering into human food chain daily. Recovery of MAP from patients with inflammatory bowel disease or Crohn's disease and animal healthcare workers suffering with chronic gastrointestinal problems indicate a close association of MAP with a number of chronic and other diseases affecting human health. Higher bioload of MAP in the animals increases the risk of exposure to the human population with MAP. This review summarizes the current status of MAP infection in animals as well as in human beings and also highlights the prospects of effective management and control of disease in animals to reduce the risk of exposure to human population.

  4. 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

    2016-07-07

    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.

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

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    Daniela Paccagnini

    2009-09-01

    Full Text Available 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.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.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.

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

    Science.gov (United States)

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

    2013-12-16

    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 20 after inoculation. MAP was not detected during their preparation and short-term storage but was recuperated after starting at 180 min after inoculation storage. Live bacterial counts of E. coli, and S. Enteritidis increased during the first 24 hours, followed by a slight decrease towards the end of the study. In this study it was shown how MAP, E. coli, and S. Enteritidis resisted the acidic conditions generated during the preparation of yogurt and low storage temperatures. This work contributes to current knowledge regarding survival of MAP, E. coli, and S. Enteritidis during preparation and refrigerated storage of yogurt and emphasizes the need to improve hygiene measures to ensure the absence of these pathogenic microorganisms in dairy products.

  7. Inhibition of phagosome maturation and survival of Mycobacterium avium subspecies paratuberculosis in polymorphonuclear leukocytes from Crohn's disease patients.

    Science.gov (United States)

    Rumsey, John; Valentine, John F; Naser, Saleh A

    2006-04-01

    Mycobacterium avium subspecies paratuberculosis (MAP) is an intracellular pathogen that is known to parasitize macrophages. MAP is the known etiological agent of Johne's disease and implicated in the etiology of Crohn's disease. In this study, the survival of human-derived MAP isolate following phagocytosis was evaluated using murine macrophage cell line J774A.1 and polymorphonuclear cells (PMNC's) from six Crohn's disease patients. PMNC's from five healthy individuals and four ulcerative colitis patients, as well as Escherichia coli and Mycobacterium tuberculosis, were included as controls (MOI 10:1). Maturation of the phagosome was determined by evaluating the presence of stage specific markers on the surface of the phagosomal membrane. The endosomal protein, transferrin receptor, and the lysosomal protein, Lamp-1, were then immunostained with Cy-5 conjugated secondary antibodies, and colocalization of bacteria with each marker was evaluated separately using confocal scanning laser microscopy (CSLM). In both models, colocalization of viable MAP and M. tuberculosis with the early endosomal marker occurred with a higher frequency than did association with the late lysosomal marker, as compared to live E. coli, and all dead bacterial species. Using differential live/dead staining and fluorescent microscopy, survival of M. tuberculosis and MAP was calculated to be 85% and 79%, respectively compared to only 14% for E. coli. Overall, MAP survival in murine macrophages and human PMNCs appears to mimic M. tuberculosis, suggesting the ability of this microorganism to resist phagolysosome fusion, by maintaining association with the early endosomes. The data supports MAP virulence in humans.

  8. 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.

  9. Cell-Mediated and Humoral Immune Responses after Immunization of Calves with a Recombinant Multiantigenic Mycobacterium avium subsp. paratuberculosis Subunit Vaccine at Different Ages

    DEFF Research Database (Denmark)

    Thakur, Aneesh; Aagaard, Claus; Stockmarr, Anders

    2013-01-01

    Neonates and juvenile ruminants are very susceptible to paratuberculosis infection. This is likely due to a high degree of exposure from their dams and an immature immune system. To test the influence of age on vaccine-induced responses, a cocktail of recombinant Mycobacterium avium subsp....... paratuberculosis proteins (MAP0217, MAP1508, MAP3701c, MAP3783, and MAP1609c/Ag85B) was formulated in a cationic liposome adjuvant (CAF01) and used to vaccinate animals of different ages. Male jersey calves were divided into three groups that were vaccinated at 2, 8, or 16 weeks of age and boosted twice at weeks 4...... and 12 relative to the first vaccination. Vaccine-induced immune responses, the gamma interferon (IFN-γ) cytokine secretion and antibody responses, were followed for 20 weeks. In general, the specific responses were significantly elevated in all three vaccination groups after the first booster...

  10. Pan-genomic analysis of bovine monocyte-derived macrophage gene expression in response to in vitro infection with Mycobacterium avium subspecies paratuberculosis

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    MacHugh David E

    2012-03-01

    Full Text Available Abstract Mycobacterium avium subspecies paratuberculosis is the causative agent of Johne’s disease, an intestinal disease of ruminants with major economic consequences. Infectious bacilli are phagocytosed by host macrophages upon exposure where they persist, resulting in lengthy subclinical phases of infection that can lead to immunopathology and disease dissemination. Consequently, analysis of the macrophage transcriptome in response to M. avium subsp. paratuberculosis infection can provide valuable insights into the molecular mechanisms that underlie Johne’s disease. Here, we investigate pan-genomic gene expression in bovine monocyte-derived macrophages (MDM purified from seven age-matched females, in response to in vitro infection with M. avium subsp. paratuberculosis (multiplicity of infection 2:1 at intervals of 2 hours, 6 hours and 24 hours post-infection (hpi. Differentially expressed genes were identified by comparing the transcriptomes of the infected MDM to the non-infected control MDM at each time point (adjusted P-value threshold ≤ 0.10. 1050 differentially expressed unique genes were identified 2 hpi, with 974 and 78 differentially expressed unique genes detected 6 and 24 hpi, respectively. Furthermore, in the infected MDM the number of upregulated genes exceeded the number of downregulated genes at each time point, with the fold-change in expression for the upregulated genes markedly higher than that for the downregulated genes. Inspection and systems biology analysis of the differentially expressed genes revealed an enrichment of genes involved in the inflammatory response, cell signalling pathways and apoptosis. The transcriptional changes associated with cellular signalling and the inflammatory response may reflect different immuno-modulatory mechanisms that underlie host-pathogen interactions during infection.

  11. 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...... 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...

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

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    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

  13. Short communication: Application of an N-acetyl-L-cysteine-NaOH decontamination method for the recovery of viable Mycobacterium avium subsp. paratuberculosis from milk of naturally infected cows

    Science.gov (United States)

    Mycobacterium avium subsp. paratuberculosis (MAP) is shed into the milk of cattle affected by Johne’s disease and, therefore, is a route of transmission for infection in youngstock in dairy herds. The objective of this study was to validate a decontamination and culture protocol for the recovery of ...

  14. 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...

  15. 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...

  16. 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.

  17. Pathogenesis, Molecular Genetics, and Genomics of Mycobacterium avium subsp. paratuberculosis, the Etiologic Agent of Johne’s Disease

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    Govardhan Rathnaiah

    2017-11-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (MAP is the etiologic agent of Johne’s disease in ruminants causing chronic diarrhea, malnutrition, and muscular wasting. Neonates and young animals are infected primarily by the fecal–oral route. MAP attaches to, translocates via the intestinal mucosa, and is phagocytosed by macrophages. The ensuing host cellular immune response leads to granulomatous enteritis characterized by a thick and corrugated intestinal wall. We review various tissue culture systems, ileal loops, and mice, goats, and cattle used to study MAP pathogenesis. MAP can be detected in clinical samples by microscopy, culturing, PCR, and an enzyme-linked immunosorbent assay. There are commercial vaccines that reduce clinical disease and shedding, unfortunately, their efficacies are limited and may not engender long-term protective immunity. Moreover, the potential linkage with Crohn’s disease and other human diseases makes MAP a concern as a zoonotic pathogen. Potential therapies with anti-mycobacterial agents are also discussed. The completion of the MAP K-10 genome sequence has greatly improved our understanding of MAP pathogenesis. The analysis of this sequence has identified a wide range of gene functions involved in virulence, lipid metabolism, transcriptional regulation, and main metabolic pathways. We also review the transposons utilized to generate random transposon mutant libraries and the recent advances in the post-genomic era. This includes the generation and characterization of allelic exchange mutants, transcriptomic analysis, transposon mutant banks analysis, new efforts to generate comprehensive mutant libraries, and the application of transposon site hybridization mutagenesis and transposon sequencing for global analysis of the MAP genome. Further analysis of candidate vaccine strains development is also provided with critical discussions on their benefits and shortcomings, and strategies to develop a highly

  18. Analysis of the bovine monocyte-derived macrophage response to Mycobacterium avium subspecies paratuberculosis infection using RNA-seq

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    Maura E Casey

    2015-02-01

    Full Text Available Johne’s disease, caused by infection with Mycobacterium avium subsp. paratuberculosis, (MAP, is a chronic intestinal disease of ruminants with serious economic consequences for cattle production in the United States and elsewhere. During infection, MAP bacilli are phagocytosed and subvert host macrophage processes, resulting in subclinical infections that can lead to immunopathology and dissemination of disease. Analysis of the host macrophage transcriptome during infection can therefore shed light on the molecular mechanisms and host-pathogen interplay associated with Johne’s disease. Here we describe results of an in vitro study of the bovine monocyte-derived macrophage (MDM transcriptome response during MAP infection using RNA-seq. MDM were obtained from seven age- and sex-matched Holstein-Friesian cattle and were infected with MAP across a six-hour infection time course with non-infected controls. We observed 245 and 574 differentially expressed genes in MAP-infected versus non-infected control samples (adjusted P value ≤ 0.05 at 2 and 6 hours post-infection, respectively. Functional analyses of these differentially expressed genes, including biological pathway enrichment, highlighted potential functional roles for genes that have not been previously described in the host response to infection with MAP bacilli. In addition, differential expression of pro- and anti-inflammatory cytokine genes, such as those associated with the IL-10 signaling pathway, and other immune-related genes that encode proteins involved in the bovine macrophage response to MAP infection emphasize the balance between protective host immunity and bacilli survival and proliferation. Systematic comparisons of RNA-seq gene expression results with Affymetrix® microarray data generated from the same experimental samples also demonstrated that RNA-seq represents a superior technology for studying host transcriptional responses to intracellular infection.

  19. The Impact of the Antimicrobial Compounds Produced by Lactic Acid Bacteria on the Growth Performance of Mycobacterium avium subsp. paratuberculosis

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    Petr Kralik

    2018-04-01

    Full Text Available Cell-free supernatants (CFSs extracted from various lactic acid bacteria (LAB cultures were applied to Mycobacterium avium subsp. paratuberculosis (MAP cells to determine their effect on MAP viability. In addition, 5% lactic acid (LA; pH 3 and commercially synthetized nisin bacteriocin were also tested. This procedure was chosen in order to mimic the influence of LAB compounds during the production and storage of fermented milk products, which can be contaminated by MAP. Its presence in milk and milk products is of public concern due to the possible ingestion of MAP by consumers and the discussed role of MAP in Crohn’s disease. Propidium monoazide real-time PCR (PMA qPCR was used for viability determination. Although all CFS showed significant effects on MAP viability, two distinct groups of CFS – effective and less effective – could be distinguished. The effective CFSs were extracted from various lactobacilli cultures, their pH values were mostly lower than 4.5, and their application resulted in >2 log10 reductions in MAP viability. The group of less effective CFS were filtered from Lactococcus and enterococci cultures, their pH values were higher than 4.5, and their effect on MAP viability was <2 log10. LA elicited a reduction in MAP viability that was similar to that of the group of less effective CFS. Almost no effect was found when using commercially synthetized nisin at concentrations of 0.1–1000 μg/ml. A combination of the influence of the type of bacteriocin, the length of its action, bacteriocin production strain, and pH are all probably required for a successful reduction in MAP viability. However, certain bacteriocins and their respective LAB strains (Lactobacillus sp. appear to play a greater role in reducing the viability of MAP than pH.

  20. The Mycobacterium avium subsp. Paratuberculosis protein MAP1305 modulates dendritic cell-mediated T cell proliferation through Toll-like receptor-4

    Science.gov (United States)

    Lee, Su Jung; Noh, Kyung Tae; Kang, Tae Heung; Han, Hee Dong; Shin, Sung Jae; Soh, Byoung Yul; Park, Jung Hee; Shin, Yong Kyoo; Kim, Han Wool; Yun, Cheol-Heui; Park, Won Sun; Jung, In Duk; Park, Yeong-Min

    2014-01-01

    In this study, we show that Mycobacterium avium subsp. Paratuberculosis MAP1305 induces the maturation of bone marrow-derived dendritic cells (BMDCs), a representative antigen presenting cell (APC). MAP1305 protein induces DC maturation and the production of pro-inflammatory cytokines (Interleukin (IL)-6), tumor necrosis factor (TNF)-α, and IL-1β) through Toll like receptor-4 (TLR-4) signaling by directly binding with TLR4. MAP1305 activates the phosphorylation of MAPKs, such as ERK, p38MAPK, and JNK, which is essential for DC maturation. Furthermore, MAP1305-treated DCs transform naïve T cells to polarized CD4+ and CD8+ T cells, thus indicating a key role for this protein in the Th1 polarization of the resulting immune response. Taken together, M. avium subsp. Paratuberculosis MAP1305 is important for the regulation of innate immune response through DC-mediated proliferation of CD4+ and CD8+ T cells. [BMB Reports 2014; 47(2): 115-120] PMID:24393523

  1. Molecular epidemiological confirmation and circumstances of occurrence of sheep (S) strains of Mycobacterium avium subsp. paratuberculosis in cases of paratuberculosis in cattle in Australia and sheep and cattle in Iceland.

    Science.gov (United States)

    Whittington, R J; Taragel, C A; Ottaway, S; Marsh, I; Seaman, J; Fridriksdottir, V

    2001-04-19

    Distinct strains of Mycobacterium avium subsp. paratuberculosis with a tendency to segregate in either sheep, or cattle and other ruminants, have been described and are known as S and C strains, respectively. These strains can be distinguished by a polymorphism in the IS1311 element and other DNA-based methods. C strains are relatively easy to culture from tissues and faeces of animals with paratuberculosis but S strains are difficult to culture. A retrospective survey of archival formalin-fixed paraffin-embedded tissue samples from culture negative Australian paratuberculous cattle was undertaken to determine whether infection in these cases was due to S strains. Polymerase chain reaction and restriction endonuclease analysis of the amplified product was used to identify the polymorphism in IS1311. Three cases of bovine paratuberculosis due to S strain were confirmed from three different farms. A serological survey led to the identification of a further two cases on one of these farms. S strains were also identified in archival tissues from paratuberculous sheep and cattle from Iceland, confirming epidemiological and microbiological evidence that paratuberculosis in Iceland was due to S strain following importation of infected sheep from Europe. In each bovine case in both Iceland and Australia there had been direct or indirect contact of calves with paratuberculous sheep. We were unable to determine whether S strains had established endemic infection in cattle or whether repeated infection from sheep had occurred. Limited epidemiological evidence suggests that transmission of S strains to cattle in Australia has been uncommon under extensive grazing conditions. In Iceland, different husbandry practices appear to have favoured transmission of S strains to cattle.

  2. Stability of genotyping target sequences of Mycobacterium avium subsp. paratuberculosis upon cultivation on different media, in vitro- and in vivo passage, and natural infection.

    Science.gov (United States)

    Kasnitz, Nadine; Köhler, Heike; Weigoldt, Mathias; Gerlach, Gerald F; Möbius, Petra

    2013-12-27

    Mycobacterium (M.) avium subsp. paratuberculosis - the causative agent of paratuberculosis (Johne's disease) - affects domestic and wild ruminants worldwide. Recently, different typing techniques have been combined to provide sufficient discriminatory power for the differentiation of isolates and for epidemiological studies. In order to challenge the reliability of this approach the stability of different M. avium subsp. paratuberculosis genotypes determined after primary isolation was investigated after sub-cultivation on six different media (A), twelve in vitro passages (B), or a singular in vivo passage (C). In addition, different isolates from a single animal or herd were investigated (D). Sub-cultures of type- and reference strains, re-isolated inoculation strain after in vivo passage, and 23 field isolates were genotyped by mycobacterial interspersed repetitive unit-variable-number of tandem-repeat (MIRU-VNTR)-, short-sequence-repeat (SSR)-, and IS900-based restriction-fragment length-polymorphism (IS900-RFLP)-analyses and compared with initial genotypes. MIRU-VNTR-alleles (at loci 292, X3, 25, 47, 7, and 32) were stable after in vitro cultivations and after animal passage. Results of SSR analysis at Locus 1 with 7G nucleotides and at Loci 8 and 9 (tri-nucleotides) were also stable. At Locus 2 9G repeats changed into 10G after goat passage. After in vitro subculture (A+B) but not after animal passage (C) IS900-RFLP-typing revealed changes of BstEII-patterns for 3 of 23 strains (including ATCC 19698). Multiple isolates from individual animals or from a single cattle herd with natural infection (D) which exhibited identical IS900-RFLP- and MIRU-VNTR- genotypes, showed different G repeat numbers at SSR locus 2. This implies strand slippage events during chromosomal duplication of bacteria in the course of bacterial spreading within hosts and herds. Consequently, SSR-Locus 2 is not suitable as genome marker for epidemiological studies. Copyright © 2013 Elsevier

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

    Directory of Open Access Journals (Sweden)

    Ana Cláudia Coelho

    2009-05-01

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

  4. Obtención y evaluación de un derivado proteico purificado de una cepa argentina de Mycobacterium avium subsp. paratuberculosis Production and evaluation of a purified protein derivative from an Argentine strain of Mycobacterium avium subsp. paratuberculosis (MAP

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    Andrea Gioffré

    2012-09-01

    Full Text Available Los derivados proteicos purificados (PPD son mezclas antigénicas no definidas obtenidas de distintas micobacterias. Los PPD bovino (PPDb y PPD aviar (PPDa son los antígenos que se emplean para evaluar la respuesta inmunitaria celular en infecciones como tuberculosis y paratuberculosis en el bovino. El PPDa comercial se produce a partir de Mycobacterium avium subsp. avium, y no a partir de la subespecie paratuberculosis. En este trabajo se seleccionó una cepa local de Mycobacterium avium subsp. paratuberculosis cuyo patrón molecular por RFLP es el más frecuente entre los aislamientos de nuestro país que han sido estudiados, y a partir de esta, se obtuvo un derivado proteico purificado: PPDj-IB. Se emplearon tanto el PPDa comercial como el PPDj-IB como antígenos en la prueba de liberación de gamma-interferón en animales de un tambo con paratuberculosis y en animales control. Aun cuando ambos PPD fueron capaces de estimular diferencialmente la liberación de la citoquina en el tambo infectado (respecto de los tambos control, no hubo diferencias significativas en los niveles de estimulación producidos y solo dos animales fueron positivos mediante el empleo de PPDj-IB. A partir del análisis por Western blot se demostró que el contenido de lipoarabinomano y del antígeno Apa/ModD era distinto en los PDD evaluados. Estas diferencias podrían explicar, en parte, las diferencias en los niveles de estimulación en términos individuales. Si bien el empleo de PPDj-IB no mejoró significativamente los resultados de la prueba de liberación de ?IFN, es importante destacar que se logró producir en el laboratorio un PPD apto para su empleo en ensayos in vitro.Purified Protein Derivatives (PPDs are non-defined antigens prepared from mycobacteria cultures. They are usually employed to evaluate the specific cellular immune response both in animals and humans. Bovine and avian PPDs are usually employed as antigens in mycobacterial infections such as

  5. Molecular epidemiology of Mycobacterium avium subsp. paratuberculosis isolated from sheep, cattle and deer on New Zealand pastoral farms.

    Science.gov (United States)

    Verdugo, Cristobal; Pleydell, Eve; Price-Carter, Marian; Prattley, Deborah; Collins, Desmond; de Lisle, Geoffrey; Vogue, Hinrich; Wilson, Peter; Heuer, Cord

    2014-12-01

    The present study aimed to describe the molecular diversity of Mycobacterium avium subsp. paratuberculosis (MAP) isolates obtained from sheep, cattle (beef and dairy) and deer farms in New Zealand. A total of 206 independent MAP isolates (15 beef cattle, 89 dairy cattle, 35 deer, 67 sheep) were sourced from 172 species-mobs (15 beef cattle, 66 dairy cattle, 31 deer, 60 sheep). Seventeen subtypes were identified, using a combination of variable number of tandem repeats (VNTR) and short sequence repeat (SSR) methods. Rarefaction analysis, analysis of molecular variance (AMOVA), Fst pairwise comparisons and proportional similarity index (PSI) were used to describe subtype population richness, genetic structure and potential associations between livestock sectors and New Zealand two main islands (North and South). The rarefaction analysis suggests a significantly higher subtype richness in dairy cattle herds when compared to the other livestock sectors. AMOVA results indicate that the main source of subtype variation is attributable to the livestock sector from which samples were sourced suggesting that subtypes are generally sector-specific. The pairwise Fst results were similar, with low Fst values for island differences within a livestock sector when compared to between sector analyses, representing a low subtype differentiation between islands. However, for a given island, potential associations were seen between dominant subtypes and specific livestock sectors. Three subtypes accounted for 76% of the isolates. The most common of these was isolated from sheep and beef cattle in the North Island, the second most frequent subtype was mainly isolated from dairy cattle (either island), while the third most common subtype was associated with deer farmed in the South Island. The PSI analysis suggests similarities in subtypes sourced from sheep and beef cattle. This contrasted with the isolates sourced from other livestock sectors, which tended to present sector

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

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    G. S. Abdellrazeq

    2014-08-01

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

  7. Distribuição de Mycobacterium avium subespécie paratuberculosis em órgãos de camundongos C57BL/6 experimentalmente infectados

    OpenAIRE

    Schwarz, David Germano Gonçalves

    2012-01-01

    Mycobacterium avium subespécie paratuberculosis (MAP) é o agente etiológico da paratuberculose, uma enfermidade que causa enterite granulomatosa crônica preferencialmente em ruminantes domésticos e silvestres. É considerada uma doença de impacto na economia, devido às perdas no rebanho e na saúde humana, uma vez que se têm indícios da possível relação de MAP com a doença de Crohn. Embora pesquisas tenham contribuído para avanços no diagnóstico de MAP, há carências de estudos relacionados à co...

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

    DEFF Research Database (Denmark)

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

    2011-01-01

    Paratuberculosis is a chronic granulomatous infection caused by Mycobacterium avium subsp. paratuberculosis (MAP) that affects multiple ruminant species causing important economic losses. Therefore, control programmes at herd and regional levels have been established worldwide and prevalence esti...

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

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    Douarre Pierre E

    2010-09-01

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

  10. Pulsed-field gel electrophoresis profile homogeneity of Mycobacterium avium subsp. paratuberculosis isolates from cattle and heterogeneity of those from sheep and goats

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    Geijo Marivi

    2007-03-01

    Full Text Available Abstract Background Mycobacterium avium subsp. paratuberculosis (Map causes paratuberculosis in animals and is suspected of causing Crohn's Disease in humans. Characterization of strains led to classify paratuberculosis isolates in two main types, cattle type strains, found affecting all host species, and sheep type strains, reported affecting mainly sheep. In order to get a better understanding of the epidemiology of paratuberculosis a large set of Map isolates obtained from different species over the last 25 years have been characterized. Five-hundred and twenty isolates from different hosts (cattle, sheep, goats, bison, deer and wild boar and origins had been cultured and typed by IS1311 restriction-endonuclease-analysis. Two-hundred and sixty-nine isolates were further characterized by pulsed-field gel electrophoresis (PFGE using SnaBI and SpeI endonucleases. Differences in strain isolation upon various media conditions were also studied. Results All bovines, 4 and 26% of Spanish sheep and goats, respectively, and the deer and wild boar studied, carried IS1311-Cattle type strains. IS1311-Sheep type encompassed 96% and 74% of Spanish sheep and goats, and all three Portuguese sheep. Thirty-seven distinct multiplex PFGE profiles were found, giving 32 novel profiles. Profiles 2-1 and 1-1 accounted for the 85% of cattle isolates. Ten distinct profiles were detected in Spanish sheep, none of them with an incidence higher than 25%. Profile 16-11 (43% and another three profiles were identified in Spanish caprine cultures. The hierarchical analysis, clustered all profiles found in cattle, "wild" hosts and some small ruminants within the same group. The other group included 11 profiles only found in Spanish sheep and goats, including Spanish pigmented profiles. Differences in growth requirements associated with isolate genotype were observed. Conclusion Cattle in Spain are infected with cattle type strains, while sheep and goats are mainly infected

  11. Use of an Individual-based Model to Control Transmission Pathways of Mycobacterium avium Subsp paratuberculosis Infection in Cattle Herds

    NARCIS (Netherlands)

    Al-Mamun, Mohammad A; Smith, Rebecca L; Schukken, Y. H.; Grohn, Y. T.

    2017-01-01

    Johne’s disease (JD) is a chronic enteric disease in cattle caused by Mycobacterium avian subsp. paratuberculosis (MAP). Eradicating JD is a difficult task due to the long incubation period of MAP, inefficient diagnostic tests, and delayed clinical signs. Effective control strategies can help

  12. Use of an Individual-based Model to Control Transmission Pathways of Mycobacterium avium Subsp. paratuberculosis Infection in Cattle Herds

    NARCIS (Netherlands)

    Mamun Hossain, Al Shaikh Abdullah; Smith, R.L.; Schukken, Y.H.; Gröhn, Y.T.

    2017-01-01

    Johne's disease (JD) is a chronic enteric disease in cattle caused by Mycobacterium avian subsp. paratuberculosis (MAP). Eradicating JD is a difficult task due to the long incubation period of MAP, inefficient diagnostic tests, and delayed clinical signs. Effective control strategies can help

  13. Characterization of the Apa antigen from M. avium subsp. paratuberculosis: a conserved Mycobacterium antigen that elicits a strong humoral response in cattle.

    Science.gov (United States)

    Gioffré, A; Echeverría-Valencia, G; Arese, A; Morsella, C; Garbaccio, S; Delgado, F; Zumárraga, M; Paolicchi, F; Cataldi, A; Romano, M I

    2009-12-15

    Johne's disease or paratuberculosis is widespread in almost all countries and remains difficult to eradicate. Nowadays, diagnosis of Mycobacterium avium subsp. paratuberculosis (MPTB) infection is one of the main concerns. In this work, we evaluated the expression, biochemical properties and antigenicity of the Apa antigen, encoded by the gene annotated as MAP1569, in the MPTB genome. We confirmed its expression in MPTB and its glycosylation by the ConA binding assay. Although the MPTB-Apa is not an immunodominant antigen, MPTB-infected cattle showed a strong humoral response to recombinant Apa by Western blot and ELISA. Milk was also a suitable sample to be tested by ELISA. We comparatively analysed the humoral cross-reactivity to the Apa from MPTB (MPTB-Apa) and the orthologue from Mycobacterium tuberculosis (MT-Apa, identical to that from Mycobacterium bovis) in both infected and control cows. Response of M. bovis- and MPTB-infected animals against MT-Apa was similar (P=0.6985) but the response of the M. bovis-infected ones to MPTB-Apa was differential, being significantly diminished (PApa stimulation in the IFNgamma release assay, we found no significant differences when compared infected herds with non-infected ones (P=0.34). This antigen, in contrast to bovine Purified Protein Derivative (PPDb), was strongly represented in avian PPD (PPDa), as shown by the recognition of BALB/c mice hyperimmune sera against MPTB-Apa by Dot-blot immunoassay. We therefore demonstrated the antigenicity of Apa in MPTB-infected animals and a differential response to the recombinant antigen when compared to M. bovis-infected animals. These traits herein described, added to the usefulness of milk samples to detect IgG anti-Apa, could be important for routine screening in dairy cattle, considering a multiantigenic approach to overcome the lack of immunodominance.

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

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    René Ramírez G.

    2013-11-01

    Full Text Available El 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 del individuo afectado. Una vez MAP es fagocitado por el macrófago bovino, tanto el macrófago como MAP activan: el uno para tratar de destruir a MAP y luego sufrir apoptosis y el otro para evadir su destrucción dentro del fagolisosoma del macrófago. El balance de dicha confrontación molecular determina el curso inicial de la infección hacia la eliminación eficiente del microorganismo o hacia el establecimiento de la infección, que culminará en los estadios III (clínico intermitente y IV (clínica terminal de la enfermedad de Johne. En la presente revisión se discuten los diferentes mecanismos moleculares por los cuales MAP evade la respuesta inmune, con énfasis en su comportamiento dentro de la vacuola fagocítica y como el agente establece mecanismos de sobrevivencia intracelular y altera la activación de los macrófagos del hospedero y de la respuesta inmune específica.

  15. Cellular composition of granulomatous lesions in gut-associated lymphoid tissues of goats during the first year after experimental infection with Mycobacterium avium subsp. paratuberculosis.

    Science.gov (United States)

    Krüger, C; Köhler, H; Liebler-Tenorio, E M

    2015-01-15

    Mycobacterium avium subsp. paratuberculosis (MAP) causes lesions in naturally and experimentally infected ruminants which greatly differ in severity, cellular composition and number of mycobacteria. Morphologically distinct lesions are already found during the clinically inapparent phase of infection. The complex local host response and number of MAP were characterized at the initial sites of lesions, organized gut-associated lymphoid tissue, in experimentally infected goats. Tissues were collected at 3, 6, 9 and 12 month post-inoculation (mpi) from goat kids that had orally received 10 times 10mg of bacterial wet mass of MAP (JII-1961). The cellular composition of lesions in Peyer's patches in the jejunum and next to the ileocecal valve was evaluated in 21 MAP-inoculated goats, where lesions were compared with unaltered tissue of six control goats. CD68+, CD4+, CD8+, γδ T lymphocytes, B lymphocytes and plasma cells, MHC class II+ and CD25+ cells were demonstrated by immunohistochemistry in serial cryostat sections. At 3 mpi, extensive granulomatous infiltrates predominated, consisting of numerous epitheloid cells admixed with many CD4 and γδ T lymphocytes. Only single MAP were detected. This indicates a strong cellular immune reaction able to control MAP infection. γδ T lymphocytes were markedly increased in this type of lesion which may reflect their important role early in the pathogenesis of paratuberculosis. At 9 and 12 mpi, divergent lesions were observed which may reflect different outcomes of host-pathogen interactions. In five goats, minimal granulomatous lesions were surrounded by extensive lymphoplasmacytic infiltrates and no MAP were detected by immunohistochemistry. This was interpreted as effective host response that was able to eliminate MAP locally. In three goats, decreased numbers of lymphocytes, but extensive granulomatous infiltrates with numerous epitheloid cells containing increased numbers of mycobacteria were seen. This shift of the

  16. Short communication: Application of an N-acetyl-L-cysteine-NaOH decontamination method for the recovery of viable Mycobacterium avium subspecies paratuberculosis from milk of naturally infected cows.

    Science.gov (United States)

    Bradner, L; Robbe-Austerman, S; Beitz, D C; Stabel, J R

    2014-01-01

    Mycobacterium avium ssp. paratuberculosis (MAP) is shed into the milk of cattle affected by Johne's disease and, therefore, is a route of transmission for infection in young stock in dairy herds. The objective of this study was to validate a decontamination and culture protocol for the recovery of MAP from individual bovine milk samples from known infected herds. Decontamination of milk samples (n = 17) with either 0.75% hexadecylpyridinium chloride for 5h or N-acetyl-L-cysteine-1.5% sodium hydroxide (NALC-1.5% NaOH) for 15 min before culture in BACTEC 12 B (Becton Dickinson, Franklin, NJ), para-JEM [Thermo Fisher Scientific (TREK Diagnostic Systems, Inc.), Cleveland, OH], and Herrold's egg yolk (HEY; Becton Dickinson) media was compared. Treatment with NALC-NaOH resulted in a lower percentage (6%) of contaminated samples than did treatment with hexadecylpyridinium chloride (47%), regardless of culture medium. The decontamination protocol (NALC-1.5% NaOH) was then applied to milk samples (n = 144) collected from cows at 7 US dairies. Recovery of viable MAP from the milk samples was low, regardless of culture medium, with recovery from 2 samples cultured in BACTEC 12 B medium, 1 sample cultured in para-JEM medium, and no viable MAP recovered on HEY medium. However, 32 cows were fecal culture positive and 13 milk samples were positive by direct PCR, suggesting that several cows were actively shedding MAP at the time of milk collection. Contamination rates were similar across media, with 39.6, 34.7, and 41.7% of samples contaminated after culture in BACTEC 12 B, para-JEM, and HEY media, respectively. Herd-to-herd variation had a major effect on sample contamination, with the percentage of contaminated samples ranging from 4 to 83%. It was concluded that decontamination of milk with NALC-1.5% NaOH before culture in BACTEC 12 B medium was the most efficacious method for the recovery of viable MAP from milk, although the ability to suppress the growth of contaminating

  17. Experimental infection of New Zealand Merino sheep with a suspension of Mycobacterium avium subspecies paratuberculosis (Map) strain Telford: Kinetics of the immune response, histopathology and Map culture.

    Science.gov (United States)

    Dukkipati, Venkata S R; Ridler, Anne L; Thompson, Keith G; Buddle, Bryce M; Hedgespeth, Barry A; Price-Carter, Marian; Begg, Douglas J; Whittington, Richard J; Gicquel, Brigitte; Murray, Alan

    2016-11-15

    A long-term study was undertaken to monitor immune responses, faecal cultures and clinical disease in sheep experimentally infected with Mycobacterium avium subspecies paratuberculosis (Map) strain Telford. New Zealand Merino lambs (N=56) were challenged with three oral doses of Map suspension. The lambs were weighed and faecal and blood samples obtained at different time-points. At 63 weeks post-challenge, surviving sheep were euthanised and samples of liver, ileo-caecal valve and mesenteric lymph node were collected for histopathology and Map culture. High IFN-γ and antibody responses were evident as early as 8 weeks post-C1 which persisted until the end of the trial. Approximately 92% of the sheep shed Map in faeces at 36 weeks post-challenge, with the prevalence decreasing to around 40% at the end of the trial. Thirteen sheep progressively lost weight and were euthanised between weeks 32 and 58 post-challenge. Nearly 58% of surviving sheep exhibited histo-pathological lesions in at least one of the three tissues sampled, while 42% showed acid-fast bacilli in at least one tissue. A positive Map culture in at least one tissue was obtained from approximately 85% of sheep. These results indicate that the three doses of Map challenge were highly effective in establishing Johne's disease in NZ Merino lambs. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  18. 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. Copyright © 2014 Elsevier Ltd. All rights reserved.

  19. Soluble BAFF Level Is Not Correlated to Mycobacterium avium Subspecies Paratuberculosis Antibodies and Increases After Interferon-β Therapy in Multiple Sclerosis Patients.

    Science.gov (United States)

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

    2016-09-01

    B cells are being recognized as one of the major players in the pathogenesis of multiple sclerosis (MS). The B cell activating factor (BAFF) system plays an essential role in B cell homeostasis and function in the periphery. Mycobacterium avium subspecies paratuberculosis (MAP) has been previously associated to MS in Sardinia. Antibodies against a MAP surface protein, MAP_2694, have been found significantly associated to MS patients, and this response was modified by interferon-β therapy. Increased BAFF levels following IFN-β therapy have been also described in MS patients. In this study, we evaluated whether soluble BAFF levels are comparable in men and women affected by MS and performed a correlation of the reported BAFF increase in MS patients under IFN-β therapy with changes of humoral response against MAP_2694. For these reasons, we investigated 44 MS patients before and after IFN-β therapy. A significant difference of BAFF levels was found between men and women with MS; moreover, we confirmed that IFN-β therapy strongly induces BAFF serum levels, but this was not related to the modification of immunological response against MAP_2694. In conclusion, our study highlights that IFN-β therapy induces the potent B cell survival factor BAFF without alterations of the humoral immune response against MAP.

  20. Mycobacterium avium subspecies paratuberculosis - an important food borne pathogen of high public health significance with special reference to India: an update.

    Science.gov (United States)

    Chaubey, Kundan Kumar; Singh, Shoor Vir; Gupta, Saurabh; Singh, Manju; Sohal, Jagdip Singh; Kumar, Naveen; Singh, Manoj Kumar; Bhatia, Ashok Kumar; Dhama, Kuldeep

    2017-12-01

    This review underlines the public health significance of 'Indian Bison Type' of Mycobacterium avium subspecies paratuberculosis (MAP) and also its potential as 'zoonotic infection'. In the absence of control programs, bio-load of MAP is increasing and if we take total population of animals (500 million plus) and human beings (1.23 billion plus) into account, the number of infected animals and human beings will run into millions in India. Our research on screening of over 26,000 domestic livestock for MAP infection using 4 different diagnostic tests (microscopy, culture, ELISA and PCR), during last 31 years has shown that the average bio-load of MAP in the livestock population of India is very high (cattle 43%, buffaloes 36%, goats 23% and sheep 41%). 'Mass screening' of 28,291 human samples between 2008-2016 revealed also high bio-load of MAP. It has been proved that MAP is not in-activated during pasteurization and therefore live bacilli are continuously reaching human population by consumption of even pasteurized milk and other milk products. Live bacilli have also been recovered from meat products and the environment thus illustrating the potential of MAP as pathogen of public health concern. However, at present, there is inadequate scientific evidence to confirm a conclusive link between MAP infection and Johne's disease in ruminants and some cases of Crohn's disease in human beings.

  1. Expression of a Gene Encoding 34.9 kDa PPE Antigen of Mycobacterium avium subsp. paratuberculosis in E. coli

    Directory of Open Access Journals (Sweden)

    Rajib Deb

    2010-01-01

    Full Text Available Mycobacterium avium subsp. paratuberculosis (Map contains PPE family antigens which are Proline and glutamic acid rich and may play important role as T cell antigens. Hence the identification and generation of antigens are necessary for immunological characterization. In the present study, the epitopic region of a unique PPE gene encoding 34.9 kDa protein from Map was amplified by polymerase chain reaction. The gene was cloned into Escherichia coli vector pQE30 UA. The recombinant plasmid designated as pQPPE was transformed into E. coli M15 and induced with IPTG revealed the high level expression of 37.1 kDa His-fusion protein (34.9 kDa PPE and 2.2 kDa His-tag, which was confirmed by immunoblotting. Recombinant PPE protein was then purified by Ni-NTA agarose chromatography. The polyclonal antiserum raised against purified recombinant PPE protein reacted with expressed 37.1 kDa His-fusion protein as well as with Map sonicate. The protein elicited significant delayed type hypersensitivity (DTH skin reaction in mice sensitized with Map. The results indicated that the recombinant PPE protein of Map was associated with cellular immune response.

  2. Low cross-reactivity of T-cell responses against lipids from Mycobacterium bovis and M. avium paratuberculosis during natural infection

    Science.gov (United States)

    Van Rhijn, Ildiko; Nguyen, Thi Kim Anh; Michel, Anita; Cooper, Dave; Govaerts, Marc; Cheng, Tan-Yun; van Eden, Willem; Moody, D. Branch; Coetzer, Jacobus A. W.; Rutten, Victor; Koets, Ad P.

    2011-01-01

    Although CD1 proteins are known to present mycobacterial lipid antigens to T cells, there is little understanding of the in vivo behavior of T cells restricted by CD1a, CD1b and CD1c, and the relative immunogenicity and immunodominance of individual lipids within the total array of lipids that comprise a bacterium. Because bovines express multiple CD1 proteins and are natural hosts of Mycobacterium bovis and Mycobacterium avium paratuberculosis (MAP), we used them as a new animal model of CD1 function. Here, we report the surprisingly divergent responses against lipids produced by these two pathogens during infection. Despite considerable overlap in lipid content, only three out of 69 animals cross-react with M. bovis and MAP total lipid preparations. The unidentified immunodominant compound of M. bovis is a hydrophilic compound, whereas the immunodominant lipid of MAP is presented by CD1b and was identified as glucose monomycolate (GMM). The preferential recognition of GMM antigen by MAP-infected cattle may be explained by the higher expression of GMM by MAP than by M. bovis. The bacterial species-specific nature of the CD1-restricted, adaptive T-cell response affects the approach to development of lipid based immunodiagnostic tests. PMID:19688747

  3. 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

  4. 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

    Paratuberculosis is a chronic infection of the intestine of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Early stage MAP infection can be detected by measuring cell-mediated immune responses using the interferon gamma (IFN-γ) assay. Whole blood samples are cultured...... 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...... 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...

  5. 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

    2015-06-19

    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

  6. 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

    2004-01-01

    -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...... cattle of 2+ years of age. Receiver-operating characteristic (ROC) analyses supported the assumption that the IFN-gamma test had a better performance than antibody tests of animals of 1+ and 2+ years of age. However, for animals of 3+ years all tests performed equally well. Application of single sampling...... 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...

  7. Predicting the Role of IL-10 in the Regulation of the Adaptive Immune Responses in Mycobacterium avium Subsp. paratuberculosis Infections Using Mathematical Models

    Science.gov (United States)

    Magombedze, Gesham; Eda, Shigetoshi; Stabel, Judy

    2015-01-01

    Mycobacterium avium subsp. paratuberculosis (MAP) is an intracellular bacterial pathogen that causes Johne’s disease (JD) in cattle and other animals. The hallmark of MAP infection in the early stages is a strong protective cell-mediated immune response (Th1-type), characterized by antigen-specific γ-interferon (IFN-γ). The Th1 response wanes with disease progression and is supplanted by a non-protective humoral immune response (Th2-type). Interleukin-10 (IL-10) is believed to play a critical role in the regulation of host immune responses to MAP infection and potentially orchestrate the reversal of Th1/Th2 immune dominance during disease progression. However, how its role correlates with MAP infection remains to be completely deciphered. We developed mathematical models to explain probable mechanisms for IL-10 involvement in MAP infection. We tested our models with IL-4, IL-10, IFN-γ, and MAP fecal shedding data collected from calves that were experimentally infected and followed over a period of 360 days in the study of Stabel and Robbe-Austerman (2011). Our models predicted that IL-10 can have different roles during MAP infection, (i) it can suppress the Th1 expression, (ii) can enhance Th2 (IL-4) expression, and (iii) can suppress the Th1 expression in synergy with IL-4. In these predicted roles, suppression of Th1 responses was correlated with increased number of MAP. We also predicted that Th1-mediated responses (IFN-γ) can lead to high expression of IL-10 and that infection burden regulates Th2 suppression by the Th1 response. Our models highlight areas where more experimental data is required to refine our model assumptions, and further test and investigate the role of IL-10 in MAP infection. PMID:26619346

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

    Directory of Open Access Journals (Sweden)

    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.

  9. Comparative efficacy of an indigenous 'inactivated vaccine' using highly pathogenic field strain of Mycobacterium avium subspecies paratuberculosis 'Bison type' with a commercial vaccine for the control of Capri-paratuberculosis in India.

    Science.gov (United States)

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

    2007-10-10

    Johne's disease (JD) is endemic in goatherds located at Central Institute for Research on Goats, Makhdoom, since 1979 and lately it has been reported from farmer's herds in equal frequencies. Despite using test and slaughter method for the control of JD for more than 25 years in these herds, incidence of JD has not been reduced. Efficacy of 'indigenous vaccine' containing native 'Bison type' genotype of Mycobacterium avium subspecies paratuberculosis (MAP) was compared with commercial vaccine using challenge studies with homologous strain of MAP. Goat kids (85) were randomly divided in to three groups. Kids were vaccinated with 1 ml of vaccine subcutaneously and Sham-immunized with 1 ml of sterile PBS. All kids except 3 in each group were challenged twice at 75- and 275-day post-vaccination (DPV). Four goats each from three groups were sacrificed at 200-day post-challenge to evaluate carcass and histopathologically for vaccine and challenge response in kids of different groups. Samples (blood, serum and fecal) were screened for LTT, ELISA and shedding of bacilli and data on live animal traits, mortality and experimental sacrifice were compared. Average body weights gained by goats in three groups at different stages of trials (0, 1-75, 76-275, 276-425 DPV) showed marked improvements in performance of vaccinated groups over 'Sham-immunized' group. Effect of vaccines against challenge became visible in terms of body weights gained at 276-425 DPV ('Bison' group gained significantly higher body weights than 'Sham-immunized'). Mortality was significantly less in two vaccinated as compared to 'Sham-immunized'. Vaccinated groups also had significant stimulation and sero-conversion for cell mediated and humoral immune response, respectively as compared to 'Sham-immunized'. Results of post-challenged fecal culture showed significant reduction in shedding of MAP in both vaccinated groups than in 'Sham-immunized'. There was significant improvement in external and internal

  10. Estimation of flock/herd-level true Mycobacterium avium subspecies paratuberculosis prevalence on sheep, beef cattle and deer farms in New Zealand using a novel Bayesian model.

    Science.gov (United States)

    Verdugo, Cristobal; Jones, Geoff; Johnson, Wes; Wilson, Peter; Stringer, Lesley; Heuer, Cord

    2014-12-01

    The study aimed to estimate the national- and island-level flock/herd true prevalence (HTP) of Mycobacterium avium subsp. paratuberculosis (MAP) infection in pastoral farmed sheep, beef cattle and deer in New Zealand. A random sample of 238 single- or multi-species farms was selected from a postal surveyed population of 1940 farms. The sample included 162 sheep flocks, 116 beef cattle and 99 deer herds from seven of 16 geographical regions. Twenty animals from each species present on farm were randomly selected for blood and faecal sampling. Pooled faecal culture testing was conducted using a single pool (sheep flocks) or two pools (beef cattle/deer herds) of 20 and 10 samples per pool, respectively. To increase flock/herd-level sensitivity, sera from all 20 animals from culture negative flocks/herds were individually tested by Pourquier(®) ELISA (sheep and cattle) or Paralisa™ (deer). Results were adjusted for sensitivity and specificity of diagnostic tests using a novel Bayesian latent class model. Outcomes were adjusted by their sampling fractions to obtain HTP estimates at national level. For each species, the posterior probability (POPR) of HTP differences between New Zealand North (NI) and South (SI) Islands was obtained. Across all species, 69% of farms had at least one species test positive. Sheep flocks had the highest HTP estimate (76%, posterior probability interval (PPI) 70-81%), followed by deer (46%, PPI 38-55%) and beef herds (42%, PPI 35-50%). Differences were observed between the two main islands of New Zealand, with higher HTP in sheep and beef cattle flocks/herds in the NI. Sheep flock HTP was 80% in the NI compared with 70% (POPR=0.96) in the SI, while the HTP for beef cattle was 44% in the NI and 38% in the SI (POPR=0.80). Conversely, deer HTP was higher in the SI (54%) than the NI (33%, POPR=0.99). Infection with MAP is endemic at high prevalence in sheep, beef cattle and deer flocks/herds across New Zealand. Copyright © 2014 Elsevier B

  11. The investigation of the truncated mbtA gene within the mycobactin cluster of Mycobacterium avium subspecies paratuberculosis as a novel diagnostic marker for real-time PCR.

    Science.gov (United States)

    de Kruijf, Marcel; Coffey, Aidan; O'Mahony, Jim

    2017-05-01

    The inability of Mycobacterium avium subspecies paratuberculosis (MAP) to produce endogenous mycobactin in-vitro is most likely due to the presence of a truncated mbtA gene within the mycobactin cluster of MAP. The main goal of this study was to investigate this unique mbtA truncation as a potential novel PCR diagnostic marker for MAP. Novel primers were designed that were located within the truncated region and the contiguous MAP2179 gene. Primers were evaluated against non-MAP isolates and no amplicons were generated. The detection limit of this mbtA-MAP2179 target was evaluated using a range of MAP DNA concentrations, MAP inoculated faecal material and 20 MAP isolates. The performance of mbtA-MAP2179 was compared to the established f57 target. The detection limits recorded for MAP K-10 DNA and from MAP K-10 inoculated faecal samples were 0.34pg and 10 4 CFU/g respectively for both f57 and mbtA-MAP2179. A detection limit of 10 3 CFU/g was recorded for both targets, but not achieved consistently. The detection limit of MAP from inoculated faecal material was successful at 10 3 CFU/g for mbtA-MAP2179 when FAM probe real-time PCR was used. A MAP cell concentration of 10 2 CFU/g was detected successfully, but again not consistently achieved. All 20 mycobacterial isolates were successfully identified as MAP by f57 and mbtA-MAP2179. Interestingly, the mbtA-MAP2179 real-time PCR assay resulted in the formation of a unique melting curve profile that contained two melting curve peaks rather than one single peak. This melting curve phenomenon was attributed towards the asymmetrical GC% distribution within the mbtA-MAP2179 amplicon. This study investigated the implementation of the mbtA-MAP2179 target as a novel diagnostic marker and the detection limits obtained with mbtA-MAP2179 were comparable to the established f57 target, making the mbtA-MAP2179 an adequate confirmatory target. Moreover, the mbtA-MAP2179 target could be implemented in multiplex real-time PCR assays and

  12. Improved Culture Medium (TiKa) for Mycobacterium avium Subspecies Paratuberculosis (MAP) Matches qPCR Sensitivity and Reveals Significant Proportions of Non-viable MAP in Lymphoid Tissue of Vaccinated MAP Challenged Animals

    DEFF Research Database (Denmark)

    Bull, Tim J.; Munshil, Tulika; Melvang, Heidi Mikkelsen

    2017-01-01

    The quantitative detection of viable pathogen load is an important tool in determining the degree of infection in animals and contamination of foodstuffs. Current conventional culture methods are limited in their ability to determine these levels in Mycobacterium avium subspecies paratuberculosis...... in recoverability and an improved sensitivity of up to three logs when compared with conventional culture. Using TiKa culture, MAP clumping was minimal and produced visible colonies in half the time required by standard culture methods. Parallel quantitative evaluation of the TiKa culture approach and qPCR on MAP......, the relative fold changes in Geq and cfu from the TiKa culture approach suggests that non-mucosal tissue loads from MAP infected animals contained a reduced proportion of non-viable MAP (mean 19-fold) which was reduced significantly further (mean 190-fold) in vaccinated "reactor" calves. This study shows Ti...

  13. 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...

  14. Bayesian estimation of sensitivity and specificity of a commercial serum/milk ELISA against the Mycobacterium avium subsp. Paratuberculosis (MAP) antibody response for each lactation stage in Greek dairy sheep.

    Science.gov (United States)

    Angelidou, Elisavet; Kostoulas, Polychronis; Leontides, Leonidas

    2016-02-01

    A total of 854 paired milk and blood samples were collected from ewes of a Greek flock and used to estimate the sensitivity and specificity of a commercial ELISA for detection of Mycobacterium avium subsp. paratuberculosis (MAP) specific antibodies in each stage of lactation. We implemented Bayesian mixture models to derive the distributions of the test response for the healthy and the infected ewes. In the colostrum period, early, mid and late lactation stage the median values of the area under the curves (AUC) were 0.61 (95% credible interval: 0.50; 0.84), 0.61 (0.51;0.84), 0.65 (0.51;0.91), 0.65(0.51;0.89) for the serum ELISA and and 0.60 (0.50; 0.84), 0.61 (0.50; 0.84), 0.67(0.51; 0.91), 0.66(0.50; 0.90) for the milk ELISA, respectively. Both serum and milk ELISA had low to average overall discriminatory ability as measured by the area under the curves and comparable sensitivities and specificities at the recommended cutoffs. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Increasing the ex vivo antigen-specific IFN-γ production in subpopulations of T cells and NKp46+ cells by anti-CD28, anti-CD49d and recombinant IL-12 costimulation in cattle vaccinated with recombinant proteins from Mycobacterium avium subspecies paratuberculosis

    DEFF Research Database (Denmark)

    Thakur, Aneesh; Riber, Ulla; Davis, William C.

    2013-01-01

    -γ secretion by CD4, CD8, γδ T cells and NK cells. Age matched male jersey calves, experimentally infected with Mycobacterium avium subsp. paratuberculosis (MAP), were vaccinated with a cocktail of recombinant MAP proteins or left unvaccinated. Vaccine induced ex vivo recall responses were measured through Ag......T cells, which encounter specific antigen (Ag), require additional signals to mount a functional immune response. Here, we demonstrate activation of signal 2, by anti-CD28 mAb (aCD28) and other costimulatory molecules (aCD49d, aCD5), and signal 3, by recombinant IL-12, enhance Ag-specific IFN...

  16. Identification of Mycobacterium avium subsp. hominissuis Isolated From Drinking Water

    Science.gov (United States)

    Mycobacterium avium (MA) is divided into four subspecies based primarily on host-range and consists of MA subsp. avium (birds), MA subsp. silvaticum (wood pigeons), MA subsp. paratuberculosis (broad, poorly-defined host range), and the recently described MA subsp. hominissuis (hu...

  17. 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. Copyright © 2014 Asian-African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.

  18. 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)

    Thirty Holstein calves were obtained from two dairy farms in central Iowa at birth and randomly assigned to one of six 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; 6) CR, vitamins A, D3, E, ...

  19. 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.

  20. Reduction of slaughter value of paratuberculosis-infected dairy cows

    DEFF Research Database (Denmark)

    Kudahl, Anne Braad; Nielsen, Søren Saxmose

    2009-01-01

    The aim of this study was to evaluate the effects of infection with Mycobacterium avium subsp. paratuberculosis (MAP) on slaughter weight and slaughter value of dairy cows. Two datasets were analyzed: 1) recordings from 1,031 cows in herds in a pilot-study to control MAP infections; and 2) record...

  1. Detection of Mycobacterium avium subspecies in the gut associated lymphoid tissue of slaughtered rabbits.

    Science.gov (United States)

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

    2015-06-11

    Rabbits are susceptible to infection by different species of the genus Mycobacterium. Particularly, development of specific lesions and isolation of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis, both subspecies of the M. avium complex, has been reported in wildlife conditions. Although, rabbit meat production worldwide is 200 million tons per year, microbiological data on this source of meat is lacking and more specifically reports of mycobacterial presence in industrially reared rabbit for human consumption have not been published. To this end, we sought mycobacteria by microbiological and histopathological methods paying special attention to Mycobacterium avium subsp. paratuberculosis in rabbits from commercial rabbitries from the North East of Spain. M. avium subsp. paratuberculosis was not detected either by culture or PCR. However, Mycobacterium avium subsp. avium was detected in 15.15% (10/66) and Mycobacterium avium subsp. hominissuis was detected in 1.51% (1/66) of gut associated lymphoid tissue of sampled animals by PCR, whereas caecal contents were negative. 9% (6/66) of the animals presented gross lesions suggestive of lymphoid activation, 6% (4/66) presented granulomatous lesions and 3% (2/66) contained acid fast bacilli. Mycobacterial isolation from samples was not achieved, although colonies of Thermoactinomycetes sp. were identified by 16s rRNA sequencing in 6% (4/66) of sampled animals. Apparently healthy farmed rabbits that go to slaughter may carry M. avium subspecies in gut associated lymphoid tissue.

  2. Cell wall peptidolipids of Mycobacterium avium: from genetic prediction to exact structure of a nonribosomal peptide

    Science.gov (United States)

    Total lipids from an M. avium subsp. paratuberculosis (Map) ovine strain (S-type) contained no identifiable glycopeptidolipids or lipopentapeptide, yet both lipids are present in other M. avium subspecies. We determined the genetic and phenotypic basis for this difference using sequence analysis and...

  3. Production effects of pathogens causing bovine leukosis, bovine viral diarrhea, paratuberculosis, and neosporosis.

    Science.gov (United States)

    Tiwari, A; Vanleeuwen, J A; Dohoo, I R; Keefe, G P; Haddad, J P; Tremblay, R; Scott, H M; Whiting, T

    2007-02-01

    The primary purpose of this research was to determine associations among seropositivity for bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV), Mycobacterium avium ssp. paratuberculosis (MAP), and Neospora caninum (NC) and each of 3 outcome variables (305-d milk, fat, and protein production) in Canadian dairy cattle. Serum samples from up to 30 randomly selected cows from 342 herds on monthly milk testing were tested for antibodies against BLV (IDEXX ELISA; IDEXX Corporation, Westbrook, ME), MAP (IDEXX or Biocor ELISA; Biocor Animal Health, Inc., Omaha, NE), and NC (IDEXX or Biovet ELISA; Biovet Inc., St. Hyacinthe, Quebec, Canada). Up to 5 unvaccinated cattle over 6 mo of age were tested for virus-neutralizing antibodies to the Singer strain of type 1 BVDV. Dairy Herd Improvement records were obtained electronically for all sampled cows. Linear mixed models with herd and cow as random variables were fit, with significant restricted maximum likelihood estimates of outcome effects being obtained, while controlling for potential confounding variables. Bovine leukemia virus seropositivity was not associated with 305-d milk, 305-d fat, or 305-d protein production. Cows in BVDV-seropositive herds (at least one unvaccinated animal with a titer > or =1:64) had reductions in 305-d milk, fat, and protein of 368, 10.2, and 9.5 kg, respectively, compared with cows in BVDV-seronegative herds. Mycobacterium avium ssp. paratuberculosis seropositivity was associated with lower 305-d milk of 212 kg in 4+-lactation cows compared with MAP-seronegative 4+-lactation cows. Neospora caninum seropositivity in primiparous cows was associated with lower 305-d milk, fat, and protein of 158, 5.5, and 3.3 kg, respectively, compared with NC-seronegative primiparous cows. There were no interactions among seropositivity for any of the pathogens and their effects on any of the outcomes examined, although the low MAP seroprevalence limited this analysis. Results from this research

  4. Progressive bovine paratuberculosis is associated with local loss of CD4(+) T cells, increased frequency of gamma delta T cells, and related changes in T-cell function

    NARCIS (Netherlands)

    Koets, A.; Rutten, V.; Hoek, van A.; Mil, van F.; Muller, K.; Bakker, D.; Gruys, E.; Eden, van W.

    2002-01-01

    Bovine paratuberculosis is caused by the infection of young calves with Mycobacterium avium subsp. paratuberculosis, resulting in a chronic granulomatous infection of predominantly the ileum. After an incubation period of 2 to 5 years, the disease becomes progressive in some of the chronically

  5. Comparison of commercial DNA extraction kits and quantitative PCR systems for better sensitivity in detecting the causative agent of paratuberculosis in dairy cow fecal samples.

    Science.gov (United States)

    Fock-Chow-Tho, D; Topp, E; Ibeagha-Awemu, E A; Bissonnette, N

    2017-01-01

    Mycobacterium avium ssp. paratuberculosis (MAP) causes ruminant paratuberculosis (Johne's disease) worldwide. Oral-fecal contamination is the most important mode of transmission of paratuberculosis, so eradicating MAP-shedding animals could prevent disease propagation. Fecal culture, a well-known method for MAP diagnosis, requires costly specialized media and a long incubation time that sometimes ends in disappointing bacterial contamination. To facilitate the efforts of control programs, we evaluated the performance of direct fecal quantitative PCR (qPCR) assays for their sensitivity and robustness for MAP detection. Commercial kits use different strategies for extracting DNA, combined with qPCR systems, to detect the presence of MAP in fecal samples. In this study, we compared the sensitivity of 3 commercially available DNA extraction kits (A, B, and C) combined with 2 qPCR systems (T and V) for the detection of MAP in infectious cows. A total of 49 dairy cows from 5 herds were sampled twice a year for 3 yr and diagnosed using fecal culture and ELISA. Eight replicates of their fecal samples from the first sampling were tested using each DNA extraction method and qPCR detection system. Although all 3 of the commercial DNA extraction kits have been previously described as very efficient for the diagnosis of paratuberculosis, kit B provided the highest sensitivity. Indeed, 89% of the cows declared positive for paratuberculosis by both fecal culture and ELISA were identified with kit B, whereas only 23 and 43% of the cows were identified with kits A and C, respectively. Interestingly, kit B was able to detect some low-MAP shedders. The qPCR detection system also played a critical role: system T yielded qPCR with the highest sensitivity. The results of this study suggest that DNA extraction kit B combined with detection system T provides the best amplification of MAP DNA from fecal samples with the highest sensitivity and specificity. Although 1 DNA extraction and q

  6. 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...

  7. Simulating the Epidemiological and Economic Impact of Paratuberculosis Control Actions in Dairy Cattle

    Directory of Open Access Journals (Sweden)

    Carsten Kirkeby

    2016-10-01

    Full Text Available We describe a new mechanistic bio-economic model for simulating the spread of Mycobacterium avium ssp. paratuberculosis (MAP within a dairy cattle herd. The model includes age-dependent susceptibility for infection; age-dependent sensitivity for detection; environmental MAP build-up in five separate areas of the farm; in utero infection; infection via colostrum and waste milk, and it allows for realistic culling (i.e. due to other diseases by including a ranking system. We calibrated the model using a unique dataset from Denmark, including 102 random farms with no control actions against spread of MAP. Likewise, four control actions recommended in the Danish MAP control program were implemented in the model based on reported management strategies in Danish dairy herds in a MAP control scheme. We tested the model parameterization in a sensitivity analysis. We show that a test-and-cull strategy is on average the most cost-effective solution to decrease the prevalence and increase the total net revenue on a farm with low hygiene, but not more profitable than no control strategy on a farm with average hygiene. Although it is possible to eradicate MAP from the farm by implementing all four control actions from the Danish MAP control program, it was not economically attractive since the expenses for the control actions outweigh the benefits. Furthermore, the three most popular control actions against the spread of MAP on the farm were found to be costly and inefficient in lowering the prevalence when used independently.

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

    DEFF Research Database (Denmark)

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

    2003-01-01

    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...... 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......-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...

  9. Bovine paratuberculosis: a review of the advantages and disadvantages of different diagnostic tests Paratuberculosis bovina: una revisión sobre las ventajas y desventajas de las diferentes pruebas diagnósticas

    OpenAIRE

    Liliana R. Gilardoni; Fernando A. Paolicchi; Silvia L. Mundo

    2012-01-01

    Paratuberculosis (PTB), or Johne's disease, is a chronic infectious granulomatous enteritis of ruminants, caused by Mycobacterium avium subspecies paratuberculosis (Map). It is characterized by diarrhea and progressive cachexia, which may cause the death of the animal. Calves are the most susceptible to infection. Infected animals excrete Map mainly by the feces. PTB is endemic worldwide, with high prevalence levels, strong economic impact and public health relevance because of its possible a...

  10. Cattle transfers between herds under paratuberculosis surveillance in The Netherlands are not random

    NARCIS (Netherlands)

    Weber, M.F.; Roermund, van H.J.W.; Vernooij, J.C.M.; Kalis, C.H.J.; Stegeman, J.A.

    2006-01-01

    The rate and structure of cattle transfers between 206 Dutch cattle herds with a 'Mycobacterium avium subsp. paratuberculosis (Map)-free' status by November 2002, were analyzed over a 3-year period (November 1999-November 2002). Of the 206 'Map-free' herds, 184 were closed herds during the period

  11. [Paratuberculosis in a miniature donkey (Equus asinus f. asinus)].

    Science.gov (United States)

    Stief, Birgit; Möbius, Petra; Türk, Heidemarie; Hörügel, Uwe; Arnold, Carina; Pöhle, Dietrich

    2012-01-01

    Paratuberculosis is mainly an infectious disease of ruminants with worldwide distribution. Infection occurs in early stages of life. Other animal species beyond ruminants are rarely affected, however, experimental and natural infections are possible. A case of paratuberculosis in a miniature donkey (Equus asinus f. asinus) with typical clinical and pathomorphological changes is reported here. Lesions were mainly observed in the intestine. Causative for the profuse diarrhoea with emaciation was massive diffuse granulomatous enteritis involving large quantities of acid-fast organism mainly in macrophages. Granulomatous inflammation with acid-fast bacilli again in macrophages to a lesser degree could be detected in the liver. Mycobacterium avium subsp. paratuberculosis (MAP) was isolated from intestinal contents after an incubation period of four weeks. MAP-specific DNA (IS900 and f57) was detected by polymerase chain reaction in culture material. Additionally MAP-isolates were characterized by multi-target genotyping (MIRU-VNTR- and MLSSR-typing). Isolates belonged to the Type II group and exhibited a unique genotype different from other MAP strains in Germany. The donkey originated from a donkey breeding farm in France with intensive free ranging cattle in the neighbourhood and could have been infected there. Donkeys should be considered as paratuberculosis-susceptible animals in exceptional cases and as possible reservoirs or disseminators of infection.

  12. Paratuberculosis in ruminants in Brasil: a review

    OpenAIRE

    Yamasaki, Elise M.; Brito, Marilene F.; Mota, Rinaldo A.; McIntosh, Douglas; Tokarnia, Carlos H.

    2013-01-01

    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...

  13. 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 uniquen......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...

  14. Paratuberculosis in a domestic dog in South Africa

    Directory of Open Access Journals (Sweden)

    Michele A. Miller

    2017-03-01

    Full Text Available This case report shows that Mycobacterium avium subsp. paratuberculosis (MAP infection can cause clinical disease in domestic dogs, and should be considered as a differential diagnosis for gastrointestinal inflammatory conditions. A male dachshund presented with lethargy and pain. Enlarged mesenteric lymph nodes were found on abdominal ultrasound examination. Cytological examination of lymph node aspirates was consistent with granulomatous inflammation, which was culture-confirmed as MAP. Although we were unable to confirm the source of infection, the dog’s history included exposure to sheep in the Western Cape.

  15. Paratuberculosis in a domestic dog in South Africa

    Directory of Open Access Journals (Sweden)

    Michele A. Miller

    2017-01-01

    Full Text Available This case report shows that Mycobacterium avium subsp. paratuberculosis (MAP infection can cause clinical disease in domestic dogs, and should be considered as a differential diagnosis for gastrointestinal inflammatory conditions. A male dachshund presented with lethargy and pain. Enlarged mesenteric lymph nodes were found on abdominal ultrasound examination. Cytological examination of lymph node aspirates was consistent with granulomatous inflammation, which was culture-confirmed as MAP. Although we were unable to confirm the source of infection, the dog’s history included exposure to sheep in the Western Cape.

  16. Spatial differences in occurrence of paratuberculosis in Danish dairy herds and in control programme participation

    DEFF Research Database (Denmark)

    Bihrmann, Kristine; Nielsen, Søren Saxmose; Toft, Nils

    2012-01-01

    Paratuberculosis is a chronic infection of economic importance to the cattle industry and a voluntary control programme is offered to Danish dairy farmers. Our objective was to evaluate spatial differences in both control programme participation and paratuberculosis prevalence in Denmark. The study...... included 4414 dairy herds: 1249 were participating in the control programme, and 1503 were tested for antibodies to Mycobacterium avium subsp. paratuberculosis (MAP). Spatial differences were evaluated by kernel smoothing, kriging, and cluster analysis. Participation was lowest among herds on the island...... Zealand (≤23%). The risk of a herd being infected with MAP was found to be high on most of Zealand, but the uncertainty of this result was large due to a limited number of tested herds. In the rest of the country, the south western part of the peninsula Jutland had the highest risk of MAP (≥91%). The risk...

  17. Milk quality assurance for paratuberculosis: simulation of within-herd infection dynamics and economicsof within-herd infection dynamics and economics

    NARCIS (Netherlands)

    Weber, M.F.; Nielen, M.; Velthuis, A.G.J.; Roermund, van H.J.W.

    2008-01-01

    bulk milk quality assurance programme for Mycobacterium avium subsp. paratuberculosis (Map) in dairy herds was simulated with a stochastic simulation model (JohneSSim). The aim of this study was to evaluate the epidemiological and economic effects of preventive management measures and various test

  18. Mycobacterium Avium Complex (MAC)

    Science.gov (United States)

    ... 30, 2014 Select a Language: Fact Sheet 514 Mycobacterium Avium Complex (MAC) WHAT IS MAC? HOW DO ... INTERACTION PROBLEMS THE BOTTOM LINE WHAT IS MAC? Mycobacterium Avium Complex (MAC) is a serious illness caused ...

  19. Pathways and Genes Associated with Immune Dysfunction in Sheep Paratuberculosis

    Science.gov (United States)

    Gossner, Anton; Watkins, Craig; Chianini, Francesca; Hopkins, John

    2017-01-01

    Multibacillary and paucibacillary paratuberculosis are both caused by Mycobacterium avium subspecies paratuberculosis. Multibacillary lesions are composed largely of infected epithelioid macrophages and paucibacillary lesions contain T cells but few bacteria. Multibacillary disease is similar to human lepromatous leprosy, with variable/high levels of antibody and a dysfunctional immune response. Animals with paucibacillary disease have high cell-mediated immunity and variable levels of antibody. This study aims to characterize the immunological dysfunction using TruSeq analysis of the ileocaecal lymph node that drains disease lesions. Immune dysfunction is highlighted by repression of TCR/CD3 genes, T cell co-receptors/co-stimulators, T cell activation and signal-transduction genes. Inflammation was an acute phase response and chronic inflammation, with little evidence of acute inflammation. The high levels of immunoglobulin and plasma cell transcripts is consistent with the anti-MAP antibody responses in paratuberculosis sheep. Also notable was the overwhelming reduction in mast cell transcripts, potentially affecting DC activation of the immune response. This study also shows that there were no fundamental differences in the gene expression patterns in multibacillary and paucibacillary disease, no shift in T cell genes from Th1 to Th2 pattern but rather an incremental decline into immune dysfunction leading to multibacillary pathology. PMID:28436433

  20. Superior protection elicited by live-attenuated vaccines in the murine model of paratuberculosis.

    Science.gov (United States)

    Ghosh, Pallab; Shippy, Daniel C; Talaat, Adel M

    2015-12-16

    Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) causes Johne's disease, a chronic enteric infection in ruminants with severe economic impact on the dairy industry in the USA and worldwide. Currently, available vaccines have limited protective efficacy against disease progression and does not prevent spread of the infection among animals. Because of their ability to elicit wide-spectrum immune responses, we adopted a live-attenuated vaccine approach based on a sigH knock-out strain of M. paratuberculosis (ΔsigH). Earlier analysis of the ΔsigH mutant in mice indicated their inadequate ability to colonize host tissues, unlike the isogenic wild-type strain, validating the role of this sigma factor in M. paratuberculosis virulence. In the present study, we evaluated the performance of the ΔsigH mutant compared to inactivated vaccine constructs in a vaccine/challenge model of murine paratuberculosis. The presented analysis indicated that ΔsigH mutant with or without QuilA adjuvant is capable of eliciting strong immune responses (such as interferon gamma-γ, IFN-γ) suggesting their immunogenicity and ability to potentially initiate effective vaccine-induced immunity. Following a challenge with virulent strains of M. paratuberculosis, ΔsigH conferred protective immunity as indicated by the reduced bacterial burden accompanied with reduced lesions in main body organs (liver, spleen and intestine) usually infected with M. paratuberculosis. More importantly, our data indicated better ability of the ΔsigH vaccine to confer protection compared to the inactivated vaccine constructs even with the presence of oil-adjuvant. Overall, our approach provides a rational basis for using live-attenuated mutant strains to develop improved vaccines that elicit robust immunity against this chronic infection. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. 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.

  2. 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.

    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 life and post-exposure vaccinated at different times after inoculation or with different vaccine constructs. In contrast to common whole-cells vaccination, the FET11 vaccine did not interfere with tests for paratuberculosis or bovine tuberculosis as no measurable antibody responses by ID Screen® ELISA...... 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...

  3. Characterization of Mycobacterium paratuberculosis by gas-liquid and thin-layer chromatography and rapid demonstration of mycobactin dependence using radiometric methods

    International Nuclear Information System (INIS)

    Damato, J.J.; Knisley, C.; Collins, M.T.

    1987-01-01

    Thirty-six Mycobacterium paratuberculosis isolates of bovine, caprine, and ovine origins were evaluated by using gas-liquid chromatography (GLC), thin-layer chromatography (TLC), and BACTEC 7H12 Middlebrook TB medium in an effort to more rapidly differentiate this group of organisms from other mycobacteria. Bacterial suspensions (0.1 ml) were inoculated by syringe into 7H12 broth containing 2 micrograms of mycobactin P per ml and control broth without mycobactin P. Cultures were incubated at 37 0 C and read daily with a BACTEC Model 301. After 8 days of incubation, the growth index readings for the test broths containing mycobactin P were twice those of the control broths without mycobactin P. Sixty-five isolates of mycobacteria other than M. paratuberculosis were also examined. No difference was noted between the growth index readings of control and mycobactin-containing broths. Except for Mycobacterium avium-Mycobacterium intracellulare, TLC studies differentiated M. paratuberculosis from the other mycobacterial species tested. The GLC data reveal that all M. paratuberculosis isolates had a distinctive peak (14A) which was not found among M. avium-M. intracellulare complex organisms. These data indicate that 7H12 radiometric broth was able to rapidly demonstrate the mycobactin dependence of M. paratuberculosis and GLC and TLC procedures were capable of rapidly differentiating this organism from the other mycobacteria studied

  4. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Evaluaci??n de nuevos adyuvantes en la vacunaci??n frente a la paratuberculosis = Evaluation of new adjuvants in the vaccination against ovine paratuberculosis

    OpenAIRE

    Reyes ??vila, Luis Ernesto

    2017-01-01

    318 p. En la presente tesis doctoral se eval??a la actividad adyuvante del Montanide?? ISA 266 y Montanide?? IMS 1312 junto a bacilos inactivados de Mycobacterium avium subsp. Paratuberculosis (Map), estudiando la reacci??n local inducida en el punto de inoculaci??n y la respuesta inmune perif??rica, compar??ndolos con el adyuvante empleado en la vacuna comercial Gudair?? (aceite mineral en doble emulsi??n estable). Se desarrollan tres experiencias en las que se vacunan corderos de 30 d??a...

  6. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  8. 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.

  9. Thermal tolerance of Mycobacterium paratuberculosis.

    Science.gov (United States)

    Sung, N; Collins, M T

    1998-03-01

    D values (decimal reduction time; the time required to kill 1 log concentration of bacteria) were determined for both human and bovine strains (Dominic, Ben, BO45, and ATCC 19698) of Mycobacterium paratuberculosis in 50 mM lactate solution (pH 6.8) and in milk at four temperatures (62, 65, 68, and 71 degrees C). Viable M. paratuberculosis organisms were quantified by a radiometric culture method (BACTEC). Thermal death curves for the M. paratuberculosis strains tested were generally linear, with R2 of > or = 0.90, but a few curves (R2, 0.80 to 0.90) were better described by a quadratic equation. The human strains (Dominic and Ben) had similar D values in milk and in lactate solution. However, D values for the bovine strains (BO45 and ATCC 19698) were significantly different depending on the menstruum. D values for low-passage clinical strains (Dominic, Ben, and BO45) were lower than those of the high-passage laboratory strain (ATCC 19698). The D value based on pooled data for clinical strains of M. paratuberculosis in milk at 71 degrees C (D71 degrees C) was 11.67 s. Pooled D62 degrees C, D65 degrees C, and D68 degrees C of clinical M. paratuberculosis strains in milk were 228.8, 47.8, and 21.8 s, respectively. The Z value (the temperature required for the decimal reduction time to traverse 1 log cycle) of clinical strains in milk was 7.11 degrees C. The D values of clumped and single M. paratuberculosis cells were not significantly different. The D values of all M. paratuberculosis strains tested were considerably higher than those published for Listeria, Salmonella, and Coxiella spp. and estimated for Mycobacterium bovis, indicating that M. paratuberculosis is more thermally tolerant. This study supports the premise that M. paratuberculosis may survive high-temperature, short-time pasteurization when the initial organism concentration is greater than 10(1) cells/ml.

  10. 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

  11. 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

  12. 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........ A 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...

  13. Journal of Genetics | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Paratuberculosis is one of the chronic granulomatous enteritis that predominantly affects ruminantsworldwide, caused by Mycobacterium avium ssp. paratuberculosis (MAP). In ruminants, microsatellite polymorphisms of the 3' untranslated region (3'UTR) of the solute carrier family 11 member A1 (SLC11A1) gene were ...

  14. Evaluation of the association of SLC11A1 gene polymorphism with ...

    Indian Academy of Sciences (India)

    2017-09-12

    Sep 12, 2017 ... Paratuberculosis is one of the chronic granulomatous enteritis that predominantly affects ruminantsworldwide, caused by Mycobacterium avium ssp. paratuberculosis (MAP). In ruminants, microsatellite polymorphisms of the 3' untranslated region (3'UTR) of the solute carrier family 11 member A1 ...

  15. Meta-analysis of two genome-wide association studies of bovine paratuberculosis.

    Science.gov (United States)

    Minozzi, Giulietta; Williams, John L; Stella, Alessandra; Strozzi, Francesco; Luini, Mario; Settles, Matthew L; Taylor, Jeremy F; Whitlock, Robert H; Zanella, Ricardo; Neibergs, Holly L

    2012-01-01

    Bovine paratuberculosis (ParaTB) also known as Johne's disease, is a contagious fatal disease resulting from infection by Mycobacterium avium subspecies paratuberculosis (MAP). Previous studies have identified loci associated with ParaTB using different measurements to define cases and controls. The objective of this study was to combine the data from two recent studies to identify genetic loci associated with MAP tissue infection and humoral immune response, defined by MAP ELISA-positive cattle, by comparing cases and control animals for one or both measures of infection. The two populations used for the association analyses were a cohort of MAP tissue infected animals and control Holstein cows from the USA and the second cohort composed of ELISA-positive and ELISA-negative Holstein cows from Italy. Altogether 1190 cattle were genotyped with the Illumina BovineSNP50 BeadChip. SNP markers were removed if the minor allele frequency 5%. Animals were removed with >5% genotyping failure. Whole genome association analyses were conducted with the GRAMMAR-CG method using two different definitions of control populations. The analyses identified several loci (Pdefinition when seeking to identify markers associated with different disease responses.

  16. Association between combinations of genetic polymorphisms and epidemiopathogenic forms of bovine paratuberculosis

    Directory of Open Access Journals (Sweden)

    Ramon A. Juste

    2018-02-01

    Full Text Available Control of major mycobacterial diseases affecting livestock is a challenging issue that requires different approaches. The use of genetic markers for improving resistance to Mycobacterium avium subsp. paratuberculosis infection in cattle has been explored as a promising population strategy We performed paratuberculosis epidemiopathogenic phenotypic and genotypic characterization involving 24 SNPs in six candidate genes (NOD2, CD209, SLC11A1, SP110, TLR2 and TLR4 on 502 slaughtered Friesian cows. In the current study, we investigate whether recently proposed paratuberculosis (PTB epidemiopathogenic (EP forms (apparently free-AF, latent-LAT and patent-PAT could be associated with some combination of these 24 SNPs. Best EP form grouping was obtained using a combination of 5 SNPs in four genes (CD209: rs210748127; SLC11A1: rs110090506; SP110: rs136859213 and rs110480812; and TLR2: rs41830058. These groups were defined according to the level of infection progression risk to patent epidemiopathogenic forms and showed the following distributions: LOWIN (low with 39 (8% cases (94.9% AF/5.1% LAT/0% PAT; LATIN (low with 17 (3% cases (5.9% AF/94.1% LAT/0% PAT; AVERIN (average with 413 (82% cases (52.1% AF/38.5% LAT/9.4% PAT and PATIN (patent with 33 (7% cases (36.4% AF/24.2% LAT/39.4% PAT. Age of slaughter was significantly higher for LATIN (88.3 months compared to AVERIN (65.3 months; p = 0.0007 and PATIN (59.1 months; p = 0.0004, and for LOWIN (73.9 months compared to PATIN (p = 0.0233, and nearly significant compared to AVERIN (p = 0.0572 These results suggest that some selected genetic polymorphisms have a potential use as markers of PTB EP forms and thus add a new tool for the control of this widespread infection.

  17. Controlling bovine paratuberculosis at a regional scale: Towards a decision modelling tool.

    Science.gov (United States)

    Beaunée, Gaël; Vergu, Elisabeta; Joly, Alain; Ezanno, Pauline

    2017-12-21

    Johne's disease (paratuberculosis), a worldwide enzootic disease of cattle caused by Mycobacterium avium subsp. paratuberculosis (Map), mainly introduced into farms by purchasing infected animals, has a large economic impact for dairy producers. Since diagnostic tests used in routine are poorly sensitive, observing Map spread in the field is hardly possible, whereas there is a need for evaluating control strategies. Our objective was to provide a modelling framework to compare the efficacy of regional control strategies combining internal biosecurity measures and testing of traded animals, against Map spread in a metapopulation of dairy cattle herds. We represented 12,857 dairy herds located in Brittany (France), based on data from 2005 to 2013, used to calibrate herd sizes and demographic rates and to define trade events in a multiscale model of Map infection dynamics. By clustering and categorical descriptive analysis of intensive simulations of this model, based on a numerical experimental design, a large panel of control measures was explored. Their efficacy was assessed on model outputs such as the prevalence and probability of extinction at the metapopulation level. In addition, we proposed a scoring for the effort required to implement control measures and prioritized control strategies based on their theoretical epidemiological efficacy. Our results clearly indicate that eradication cannot be achieved on the mid term using available control measures. However, we identified relevant combinations of measures that lead to the control of Map spread with realistic level of implementation and coverage. The study highlights the challenge of controlling paratuberculosis in an endemically infected region as related to the poor test characteristics and frequent trade movements. Our model lays the foundations for a flexible and efficient tool to help collective animal health managers in defining relevant control strategies at a regional scale, accounting for local

  18. Bovine paratuberculosis: a review of the advantages and disadvantages of different diagnostic tests Paratuberculosis bovina: una revisión sobre las ventajas y desventajas de las diferentes pruebas diagnósticas

    Directory of Open Access Journals (Sweden)

    Liliana R. Gilardoni

    2012-09-01

    Full Text Available Paratuberculosis (PTB, or Johne's disease, is a chronic infectious granulomatous enteritis of ruminants, caused by Mycobacterium avium subspecies paratuberculosis (Map. It is characterized by diarrhea and progressive cachexia, which may cause the death of the animal. Calves are the most susceptible to infection. Infected animals excrete Map mainly by the feces. PTB is endemic worldwide, with high prevalence levels, strong economic impact and public health relevance because of its possible association with Crohn's disease. Although the current reference diagnostic test is identification of Map in the bacterial culture, there are different diagnostic tests to identify infected individuals and/or herds. The sensitivity and specificity of these tests vary according to the stage of the disease in the animals to be evaluated. The correct choice and application of each of these diagnostic tests will ensure their success and may allow to establish a control program. The aim of this work is to review and discuss the different diagnostic tests used in the detection of Map-infected animals, focusing on their advantages and disadvantages.La paratuberculosis (PTBC o enfermedad de Johne es una enteritis granulomatosa crónica de rumiantes, causada por Mycobacterium avium subsp. paratuberculosis (Map. Se caracteriza por producir diarrea y caquexia progresiva, la cual conduce a la muerte del animal. Los terneros son los animales más proclives a la infección. Los animales infectados excretan Map, principalmente por las heces. La PTBC es una enfermedad endémica a nivel mundial, con altos niveles de prevalencia, fuerte impacto económico e importancia en salud pública, debido a su posible asociación con la enfermedad de Crohn. Aunque la prueba de referencia diagnóstica es la identificación de Map en cultivo bacteriológico, existen diferentes pruebas diagnósticas para detectar animales o rodeos infectados. La sensibilidad y especificidad de estas pruebas

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

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    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

  20. Isolation of Mycobacterium paratuberculosis from apparently ...

    African Journals Online (AJOL)

    chronic diarrhea M.paratuberculosis was identified easily by fecal direct smear stained with ZN (Godfroid et al., 2000). In clinically affected animals with his- tological lesions in the intestines, examination of feces for bacilli in Ziehl-Neel- son stained smears will identify up to 57% of infected animals (Greig, 2000). In our case ...

  1. 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

  2. Drug susceptibility testing of Mycobacterium Avium subsp. Avium isolates from naturally infected domestic pigeons to avian tuberculosis

    Directory of Open Access Journals (Sweden)

    Kaveh Parvandar

    2016-01-01

    Conclusion: We suggest drug susceptibility testing for more nontuberculous mycobateria, particularly M. avium complex isolated from infected birds and humans, as well as molecular basics of drug sensitivity in order to detect resistance genes of pathogenic M. avium subsp. avium.

  3. Meta-analysis of two genome-wide association studies of bovine paratuberculosis.

    Directory of Open Access Journals (Sweden)

    Giulietta Minozzi

    Full Text Available BACKGROUND: Bovine paratuberculosis (ParaTB also known as Johne's disease, is a contagious fatal disease resulting from infection by Mycobacterium avium subspecies paratuberculosis (MAP. Previous studies have identified loci associated with ParaTB using different measurements to define cases and controls. The objective of this study was to combine the data from two recent studies to identify genetic loci associated with MAP tissue infection and humoral immune response, defined by MAP ELISA-positive cattle, by comparing cases and control animals for one or both measures of infection. METHODOLOGY/PRINCIPAL FINDINGS: The two populations used for the association analyses were a cohort of MAP tissue infected animals and control Holstein cows from the USA and the second cohort composed of ELISA-positive and ELISA-negative Holstein cows from Italy. Altogether 1190 cattle were genotyped with the Illumina BovineSNP50 BeadChip. SNP markers were removed if the minor allele frequency 5%. Animals were removed with >5% genotyping failure. Whole genome association analyses were conducted with the GRAMMAR-CG method using two different definitions of control populations. CONCLUSION/SIGNIFICANCE: The analyses identified several loci (P<5 e-05 associated with ParaTB, defined by positive ELISA and presence of bacteria in tissue compared to ELISA and tissue negative animals, on chromosomes 1, 12 and 15 and one unassigned SNP. These results confirmed associations on chromosome 12 and the unassigned SNP with ParaTB which had been found in the Italian population alone. Furthermore, several additional genomic regions were found associated with ParaTB when ELISA and tissue positive animals were compared with tissue negative samples. These loci were on chromosomes 1, 6, 7, 13, 16, 21,23 and 25 (P<5 e-05. The results clearly indicate the importance of the phenotype definition when seeking to identify markers associated with different disease responses.

  4. Inactivation of Mycobacterium paratuberculosis in cows' milk at pasteurization temperatures.

    Science.gov (United States)

    Grant, I R; Ball, H J; Neill, S D; Rowe, M T

    1996-02-01

    The thermal inactivation of 11 strains of Mycobacterium paratuberculosis at pasteurization temperatures was investigated. Cows' milk inoculated with M. paratuberculosis at two levels (10(7) and 10(4) CFU/ml) was pasteurized in the laboratory by (i) a standard holder method (63.5 degrees C for 30 min) and (ii) a high-temperature, short-time (HTST) method (71.7 degrees C for 15 s). Additional heating times of 5, 10, 15, 20, and 40 min at 63.5 degrees C were included to enable the construction of a thermal death curve for the organism. Viability after pasteurization was assessed by culture on Herrold's egg yolk medium containing mycobactin J (HEYM) and in BACTEC Middlebrook 12B radiometric medium supplemented with mycobactin J and sterile egg yolk emulsion. Confirmation of acid-fast survivors of pasteurization as viable M. paratuberculosis cells was achieved by subculture on HEYM to indicate viability coupled with PCR using M. paratuberculosis-specific 1S900 primers. When milk was initially inoculated with 10(6) to 10(7) CFU of M. paratuberculosis per ml, M. paratuberculosis cells were isolated from 27 of 28 (96%) and 29 of 34 (85%) pasteurized milk samples heat treated by the holder and HTST methods, respectively. Correspondingly, when 10(3) to 10(4) CFU of M. paratuberculosis per ml of milk were present before heat treatment, M. paratuberculosis cells were isolated from 14 of 28 (50%) and 19 of 33 (58%) pasteurized milk samples heat treated by the holder and HTST methods, respectively. The thermal death curve for M. paratuberculosis was concave in shape, exhibiting a rapid initial death rate followed by significant "tailing." Results indicate that when large numbers of M. paratuberculosis cells are present in milk, the organism may not be completely inactivated by heat treatments simulating holder and HTST pasteurization under laboratory conditions.

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

    NARCIS (Netherlands)

    Slater, Noa; Mitchell, Rebecca Mans; Whitlock, Robert H.; Fyock, Terry; Pradhan, Abani Kumar; Knupfer, Elena; Schukken, Ynte Hein|info:eu-repo/dai/nl/075051907; 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

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

    NARCIS (Netherlands)

    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

  7. Epidemiological and economic consequences of purchasing livestock infected with Mycobacterium avium subsp. paratuberculosis

    DEFF Research Database (Denmark)

    Kirkeby, Carsten Thure; Græsbøll, Kaare; Nielsen, Søren Saxmose

    2017-01-01

    of the infection. One strategy to control PTB in a herd is to purchase animals from farms with a low risk of MAP infection. We wanted to investigate the epidemiological and economic consequences of buying livestock from different supplier farms of low, medium or high risk, as well as farms with unknown status. We...... one, five or ten infected animals annually into an initially infection-free herd was also modeled. Our findings show that once infected, a farm can relatively safely purchase animals from other low and medium-risk farms without experiencing an increase in the prevalence, highlighting the importance...... of certification programmes. Furthermore, farms free of MAP are highly susceptible and cannot purchase more than a small number of animals per year without having a high risk of being infected. The probability of spontaneous fadeout after 10 years was 82% when introducing a single infected animal into an initially...

  8. Knowledge gaps that hamper prevention and control of Mycobacterium avium subspecies paratuberculosis infection

    DEFF Research Database (Denmark)

    Barkema, H W; Orsel, K; Nielsen, S S

    2018-01-01

    programmes are typically evaluated in a limited number of herds and the duration of the study is less than 5 year, making it difficult to adequately assess the efficacy of control programmes. In this manuscript, we identify the most important gaps in knowledge hampering JD prevention and control programmes......, including vaccination and diagnostics. Secondly, we discuss directions that research should take to address those knowledge gaps....

  9. Mycobacterium avium subsp paratuberculosis cells are surprisingly resistant to ensiling process.

    Science.gov (United States)

    Silage is a valuable source of nutrients for dairy and beef cattle in non-forage months. The most commonly ensiled crops include corn and grass forage, both of which are often fertilized with livestock manure spread by broadcasting onto the soil or by spray irrigation. Pathogen contamination may res...

  10. 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 ...

  11. Mycobacterium paratuberculosis Zoonosis-The Hundred Year War –Beyond Crohn’s Disease

    Directory of Open Access Journals (Sweden)

    Leonardo A Sechi

    2015-03-01

    Full Text Available The factitive role of Mycobacterium avium ss. paratuberculosis (MAP in Crohn’s disease has been debated for more than a century. The controversy is due to the fact that Crohn’s disease is so similar to a disease of MAP-infected ruminant animals, Johne’s disease; and, though MAP can be readily detected in the infected ruminants, it is much more difficult to detect in humans. Molecular techniques that can detect MAP in pathologic Crohn’s specimens as well as dedicated specialty labs successful in culturing MAP from Crohn’s patients have provided strong argument for MAP’s role in Crohn’s disease. Perhaps more incriminating for MAP as a zoonotic agent is the increasing number of diseases with which MAP has been related: Blau syndrome, type 1 diabetes, Hashimoto thyroiditis and multiple sclerosis. In this article we debate about genetic susceptibility to mycobacterial infection and human exposure to MAP; moreover, it suggests that molecular mimicry between protein epitopes of MAP and human proteins is a likely bridge between infection and these autoimmune disorders.

  12. M. paratuberculosis Heat Shock Protein 65 and Human Diseases: Bridging Infection and Autoimmunity

    Directory of Open Access Journals (Sweden)

    Coad Thomas Dow

    2012-01-01

    Full Text Available Mycobacterium avium subspecies paratuberculosis (MAP is the known infectious cause of Johne’s disease, an enteric inflammatory disease mostly studied in ruminant animals. MAP has also been implicated in the very similar Crohn’s disease of humans as well as sarcoidosis. Recently, MAP has been associated with juvenile sarcoidosis (Blau syndrome, autoimmune diabetes, autoimmune thyroiditis, and multiple sclerosis. While it is intuitive to implicate MAP in granulomatous diseases where the microbe participates in the granuloma, it is more difficult to assign a role for MAP in diseases where autoantibodies are a primary feature. MAP may trigger autoimmune antibodies via its heat shock proteins. Mycobacterial heat shock protein 65 (HSP65 is an immunodominant protein that shares sequential and conformational elements with several human host proteins. This molecular mimicry is the proposed etiopathology by which MAP stimulates autoantibodies associated with autoimmune (type 1 diabetes, autoimmune (Hashimoto’s thyroiditis, and multiple sclerosis. This paper proposes that MAP is a source of mycobacterial HSP65 and acts as a trigger of autoimmune disease.

  13. Significant reduction in bacterial shedding and improvement in milk production in dairy farms after the use of a new inactivated paratuberculosis vaccine in a field trial

    Directory of Open Access Journals (Sweden)

    Vazquez Patricia

    2009-11-01

    Full Text Available Abstract Background Paratuberculosis vaccination has been in use in some regions for many decades, but results have not been widely spread. A new Mycobacterium avium subsp. paratuberculosis (MAP killed vaccine was studied in relationship with its effects on fecal shedding and milk production in four farms while other two were kept as controls submitted to a test and cull scheme. Findings Fecal detection (n = 1829 and milking records (n = 2413 have been analyzed after two (5 herds and four (1 herd years of the beginning of the intervention. Shedder prevalence was reduced by 100% in three of the four vaccinated farms, 68% in the total of vaccinated animals and 46% in the two control farms. Total amount of MAP shed was reduced 77% in the vaccinated farms and 94% in the control farms. Overall milk production increased up to 3.9% after vaccination, while there was no significant difference in production after intervention in the non-vaccinated farms. Conclusion MAP shedding reduction can be quickly accomplished both by vaccination and by testing and culling. However, vaccination appears to be a less expensive and more sustainable strategy since it required one single intervention and was also associated with an increase in milk production.

  14. Lactobacillus delbrueckii ssp. lactis and ssp. bulgaricus: a chronicle of evolution in action.

    Science.gov (United States)

    El Kafsi, Hela; Binesse, Johan; Loux, Valentin; Buratti, Julien; Boudebbouze, Samira; Dervyn, Rozenn; Kennedy, Sean; Galleron, Nathalie; Quinquis, Benoît; Batto, Jean-Michel; Moumen, Bouziane; Maguin, Emmanuelle; van de Guchte, Maarten

    2014-05-28

    Lactobacillus delbrueckii ssp. lactis and ssp. bulgaricus are lactic acid producing bacteria that are largely used in dairy industries, notably in cheese-making and yogurt production. An earlier in-depth study of the first completely sequenced ssp. bulgaricus genome revealed the characteristics of a genome in an active phase of rapid evolution, in what appears to be an adaptation to the milk environment. Here we examine for the first time if the same conclusions apply to the ssp. lactis, and discuss intra- and inter-subspecies genomic diversity in the context of evolutionary adaptation. Both L. delbrueckii ssp. show the signs of reductive evolution through the elimination of superfluous genes, thereby limiting their carbohydrate metabolic capacities and amino acid biosynthesis potential. In the ssp. lactis this reductive evolution has gone less far than in the ssp. bulgaricus. Consequently, the ssp. lactis retained more extended carbohydrate metabolizing capabilities than the ssp. bulgaricus but, due to high intra-subspecies diversity, very few carbohydrate substrates, if any, allow a reliable distinction of the two ssp. We further show that one of the most important traits, lactose fermentation, of one of the economically most important dairy bacteria, L. delbruecki ssp. bulgaricus, relies on horizontally acquired rather than deep ancestral genes. In this sense this bacterium may thus be regarded as a natural GMO avant la lettre. The dairy lactic acid producing bacteria L. delbrueckii ssp. lactis and ssp. bulgaricus appear to represent different points on the same evolutionary track of adaptation to the milk environment through the loss of superfluous functions and the acquisition of functions that allow an optimized utilization of milk resources, where the ssp. bulgaricus has progressed further away from the common ancestor.

  15. MYCOBACTERIUM AVIUM AND DRINKING WATER WHAT ARE THE CONNECTIONS?

    Science.gov (United States)

    Background: Human Mycobacterium avium infections are only known to be acquired from environmental sources such as water and soil. We compared M. avium isolates from clinical and drinking water sources using molecular tools. Methods: M. avium was isolated from water samples colle...

  16. Author Details

    African Journals Online (AJOL)

    Study on grosstesticular abnormalities of rams and bucks at Luna export abattoir, Modjo, Ethiopia Abstract · Vol 59, No 3 (2011) - Articles Bovine Mastitis in Dairy Cows in Mekele, Northern Ethiopia Abstract · Vol 62, No 1 (2014) - Articles Seroprevalence of Mycobacterium avium SSP paratuberculosis infection in Ethiopian ...

  17. Managing control programs for ovine caseous lymphadenitis and paratuberculosis in Australia, and the need for persistent vaccination

    Directory of Open Access Journals (Sweden)

    Windsor PA

    2014-03-01

    Full Text Available Peter Andrew WindsorFaculty of Veterinary Science, University of Sydney, Camden, NSW, AustraliaAbstract: Ovine caseous lymphadenitis (CLA and ovine Johne's disease (OJD or paratuberculosis have been serious diseases in the Australian sheep industry, mainly causing losses from abattoir condemnations from CLA or mortalities on the farm from OJD. CLA is now a disease of minimal concern, with clinical cases reported rarely. Although OJD continues to spread through parts of the sheep population, the catastrophic losses in flocks occurring prior to the introduction of vaccination are now uncommon. Change-management factors relevant to the improvements in both prevalence and producer concerns for CLA and OJD were examined, including drivers and motivation for change, resistance to change, knowledge management, farming system dimensions and leadership. Although extension programs addressing disease risk factors are likely to be of relevance to improved knowledge and attitudes towards disease risk management of producers, improvements in disease-control practices were considered largely attributable to the introduction of vaccination programs for CLA in 1983 and OJD in 2002. Inclusion of the CLA antigen within clostridial vaccines (“6 in 1” vaccine enabled routine annual CLA vaccination to occur in an increasing proportion of the national flock, with estimates of CLA prevalence suggesting a decline from 26% in 1995 to 5.2% in 2009. Encouraging the routine vaccination of lambs for OJD (Gudair vaccine in infected flocks to reduce or avoid losses significantly reduced the within-flock prevaccination–postvaccination median prevalence from 2.72% to 0.72%, based on estimated shedding rates of Mycobacterium avium subsp. paratuberculosis determined by pooled fecal culture in 37 infected flocks vaccinating for at least 5 years. Although persistent use of CLA vaccine is a convenient intervention for producers, promoting the persistent use of OJD vaccination

  18. 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.

  19. Specificity of two tests for the early diagnosis of bovine paratuberculosis based on cell-mediated immunity : the Johnin skin test and the gamma interferon assay

    NARCIS (Netherlands)

    Kalis, CHJ; Collins, MT; Hesselink, JW; Barkema, H.W.

    2003-01-01

    Paratuberculosis in cattle is a chronic debilitating infectious disease caused by Mycobacterium paratuberculosis. Control of paratuberculosis is based on tests that principally detect advanced stages of infections: faecal culture and serology. Tests measuring cell-mediated immunity (CMI) could

  20. Technical Analysis of SSP-21 Protocol

    Energy Technology Data Exchange (ETDEWEB)

    Bromberger, S. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2017-06-09

    As part of the California Energy Systems for the Twenty-First Century (CES-21) program, in December 2016 San Diego Gas and Electric (SDG&E) contracted with Lawrence Livermore National Laboratory (LLNL) to perform an independent verification and validation (IV&V) of a white paper describing their Secure SCADA Protocol for the Twenty-First Century (SSP-21) in order to analyze the effectiveness and propriety of cryptographic protocol use within the SSP-21 specification. SSP-21 is designed to use cryptographic protocols to provide (optional) encryption, authentication, and nonrepudiation, among other capabilities. The cryptographic protocols to be used reflect current industry standards; future versions of SSP-21 will use other advanced technologies to provide a subset of security services.

  1. Labdane diterpenoids from Marrubium globosum ssp. globosum.

    Science.gov (United States)

    Takeda, Y; Yanagihara, K; Masuda, T; Otsuka, H; Honda, G; Takaishi, Y; Sezik, E; Yesilada, E

    2000-08-01

    A new labdane type diterpenoid, marrubiglobosin, was isolated from the aerial parts of Marrubium globosum ssp. globosum together with the known diterpenoids, marrubiin and marrubinone B. The structure of this new compound was elucidated by spectral methods.

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

    Directory of Open Access Journals (Sweden)

    Liliana Rosa Gilardoni

    2016-06-01

    Full Text Available Abstract 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 101 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 (<0.20, and the proportions of agreement were ≤0.55. The IMS-IS1 PCR method detected Map in 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.

  3. Tuberculosis Detection in Paratuberculosis Vaccinated Calves: New Alternatives against Interference

    Science.gov (United States)

    Serrano, Miriam; Elguezabal, Natalia; Sevilla, Iker A.; Geijo, María V.; Molina, Elena; Arrazuria, Rakel; Urkitza, Alfonso; Jones, Gareth J.; Vordermeier, Martin; Garrido, Joseba M.; Juste, Ramón A.

    2017-01-01

    Paratuberculosis vaccination in cattle has been restricted due to its possible interference with the official diagnostic methods used in tuberculosis eradication programs. To overcome this drawback, new possibilities to detect Mycobacterium bovis infected cattle in paratuberculosis vaccinated animals were studied under experimental conditions. Three groups of 5 calves each were included in the experiment: one paratuberculosis vaccinated group, one paratuberculosis vaccinated and M. bovis infected group and one M. bovis infected group. The performance of the IFN-gamma release assay (IGRA) and the skin test using conventional avian and bovine tuberculins (A- and B-PPD) but also other more specific antigens (ESAT-6/CFP10 and Rv3615c) was studied under official and new diagnostic criteria. Regarding the IGRA of vaccinated groups, when A- and B-PPD were used the sensitivity reached 100% at the first post-challenge sampling, dropping down to 40–80% in subsequent samplings. The sensitivity for the specific antigens was 80–100% and the specificity was also improved. After adapting the diagnostic criteria for the conventional antigens in the skin test, the ability to differentiate between M. bovis infected and non-infected animals included in paratuberculosis vaccinated groups was enhanced. Taking for positive a relative skin thickness increase of at least 100%, the single intradermal test specificity and sensitivity yielded 100%. The comparative intradermal test was equally accurate considering a B-PPD relative skin increase of at least 100% and greater than or equal to that produced by A-PPD. Using the specific antigens as a proteic cocktail, the specificity and sensitivity reached 100% considering the new relative and absolute cut-offs in all experimental groups (Δ≥30% and Δmm ≥ 2, respectively). Results suggest that the interference caused by paratuberculosis vaccination in cattle could be completely overcome by applying new approaches to the official

  4. Tuberculosis Detection in Paratuberculosis Vaccinated Calves: New Alternatives against Interference.

    Directory of Open Access Journals (Sweden)

    Miriam Serrano

    Full Text Available Paratuberculosis vaccination in cattle has been restricted due to its possible interference with the official diagnostic methods used in tuberculosis eradication programs. To overcome this drawback, new possibilities to detect Mycobacterium bovis infected cattle in paratuberculosis vaccinated animals were studied under experimental conditions. Three groups of 5 calves each were included in the experiment: one paratuberculosis vaccinated group, one paratuberculosis vaccinated and M. bovis infected group and one M. bovis infected group. The performance of the IFN-gamma release assay (IGRA and the skin test using conventional avian and bovine tuberculins (A- and B-PPD but also other more specific antigens (ESAT-6/CFP10 and Rv3615c was studied under official and new diagnostic criteria. Regarding the IGRA of vaccinated groups, when A- and B-PPD were used the sensitivity reached 100% at the first post-challenge sampling, dropping down to 40-80% in subsequent samplings. The sensitivity for the specific antigens was 80-100% and the specificity was also improved. After adapting the diagnostic criteria for the conventional antigens in the skin test, the ability to differentiate between M. bovis infected and non-infected animals included in paratuberculosis vaccinated groups was enhanced. Taking for positive a relative skin thickness increase of at least 100%, the single intradermal test specificity and sensitivity yielded 100%. The comparative intradermal test was equally accurate considering a B-PPD relative skin increase of at least 100% and greater than or equal to that produced by A-PPD. Using the specific antigens as a proteic cocktail, the specificity and sensitivity reached 100% considering the new relative and absolute cut-offs in all experimental groups (Δ≥30% and Δmm ≥ 2, respectively. Results suggest that the interference caused by paratuberculosis vaccination in cattle could be completely overcome by applying new approaches to the

  5. Detection of Mycobacterium avium in pet birds

    Science.gov (United States)

    Godoy, Silvia Neri; Sakamoto, Sidnei Miyoshi; de Paula, Cátia Dejuste; Catão-Dias, José Luiz; Matushima, Eliana Reiko

    2009-01-01

    The present study is a report on the presence of Mycobacterium avium in four birds of the psittaciform order kept as pets. Anatomopathological diagnosis showed lesions suggestive of the agent and presence of alcohol-acid resistant bacilli (AARB) shown by the Ziehl-Neelsen staining. The identification of Mycobacterium avium was performed by means of PRA (PCR Restriction Analysis). DNA was directly extracted from tissue of the lesions and blocked in paraffin. The role of this agent in pet bird infection is discussed, as well as its zoonotic potential. PMID:24031356

  6. Inactivation of Mycobacterium paratuberculosis in cows' milk at pasteurization temperatures.

    OpenAIRE

    Grant, I R; Ball, H J; Neill, S D; Rowe, M T

    1996-01-01

    The thermal inactivation of 11 strains of Mycobacterium paratuberculosis at pasteurization temperatures was investigated. Cows' milk inoculated with M. paratuberculosis at two levels (10(7) and 10(4) CFU/ml) was pasteurized in the laboratory by (i) a standard holder method (63.5 degrees C for 30 min) and (ii) a high-temperature, short-time (HTST) method (71.7 degrees C for 15 s). Additional heating times of 5, 10, 15, 20, and 40 min at 63.5 degrees C were included to enable the construction o...

  7. 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...... the models are somewhat different in their underlying principles and do put slightly different values on the different strategies, their overall findings are similar. Therefore, simulation models may be useful in planning paratuberculosis strategies in dairy herds, although as with all models caution...

  8. Predicting fadeout versus persistence of paratuberculosis in a dairy cattle herd for management and control purposes: a modelling study

    Directory of Open Access Journals (Sweden)

    Marcé Clara

    2011-02-01

    Full Text Available Abstract Epidemiological models enable to better understand the dynamics of infectious diseases and to assess ex-ante control strategies. For Mycobacterium avium subsp. paratuberculosis (Map, possible transmission routes have been described, but Map spread in a herd and the relative importance of the routes are currently insufficiently understood to prioritize control measures. We aim to predict early after Map introduction in a dairy cattle herd whether infection is likely to fade out or persist, when no control measures are implemented, using a modelling approach. Both vertical transmission and horizontal transmission via the ingestion of colostrum, milk, or faeces present in the contaminated environment were modelled. Calf-to-calf indirect transmission was possible. Six health states were represented: susceptible, transiently infectious, latently infected, subclinically infected, clinically affected, and resistant. The model was partially validated by comparing the simulated prevalence with field data. Housing facilities and contacts between animals were specifically considered for calves and heifers. After the introduction of one infected animal in a naive herd, fadeout occurred in 66% of the runs. When Map persisted, the prevalence of infected animals increased to 88% in 25 years. The two main transmission routes were via the farm's environment and in utero transmission. Calf-to-calf transmission was minor. Fadeout versus Map persistence could be differentiated with the number of clinically affected animals, which was rarely above one when fadeout occurred. Therefore, early detection of affected animals is crucial in preventing Map persistence in dairy herds.

  9. 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.

  10. ELECTROPHORETIC MOBILITY OF MYCOBACTERIUM AVIUM COMPLEX ORGANISMS

    Science.gov (United States)

    The electrophoretic mobilities (EPMs) of thirty Mycobacterium avium Complex (MAC) organisms were measured. The EPMs of fifteen clinical isolates ranged from -1.9 to -5.0 µm cm V-1s-1, and the EPMs of fifteen environmental isolates ranged from -1...

  11. ORF Sequence: NC_002944 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available ALLAHKGIRGLVVCCDECQQDHYHDWDMLRANLLQLLIDGTVRPHEPAYDPEPDAYVTWDYCRGYADASLNEATSDADGFHRRH ...[Mycobacterium avium subsp. paratuberculosis str. k10] MRDHLPPGLPPDPFADDPCDPSAALDAVEPGQPLDQQERMAVEADLADLAVYE

  12. Stochastic models to simulate paratuberculosis in dairy herds

    DEFF Research Database (Denmark)

    Nielsen, S.S.; Weber, M.F.; Kudahl, Anne Margrethe Braad

    2011-01-01

    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 while...

  13. Experimental Inoculation of BFDV-Positive Budgerigars (Melopsittacus undulatus with Two Mycobacterium avium subsp. avium Isolates

    Directory of Open Access Journals (Sweden)

    Aleksandra Ledwoń

    2014-01-01

    Full Text Available Beak and feather disease virus- (BFDV- positive (naturally infected but clinically healthy budgerigars (Melopsittacus undulatus were inoculated with two isolates of Mycobacterium avium subsp. avium isolated from naturally infected golden pheasant (Chrysolophus pictus and peafowl (Pavo cristatus. During a period of more than two months after inoculation, samples of cloacal and crop swabs, faeces, and blood were obtained for BFDV and Mycobacterium avium testing with PCR. Birds were euthanized nine weeks after inoculation. All infected budgerigars developed signs typical of mycobacteriosis, but more advanced clinical and pathological changes were visible in the group infected with the pheasant isolate. Only a few cloacal and crop swab samples were positive for Mycobacterium avium subsp. avium despite advanced pathological changes in the internal organs. In the groups infected with mycobacterium isolates the frequency of BFDV-positive samples was higher than in the control group. In the infected groups the frequency of BFDV was substantially higher in the cloacal swabs of birds inoculated with the pheasant isolate than in the peafowl-isolate-infected group.

  14. Experimental inoculation of BFDV-positive budgerigars (Melopsittacus undulatus) with two Mycobacterium avium subsp. avium isolates.

    Science.gov (United States)

    Ledwoń, Aleksandra; Sapierzyński, Rafał; Augustynowicz-Kopeć, Ewa; Szeleszczuk, Piotr; Kozak, Marcin

    2014-01-01

    Beak and feather disease virus- (BFDV-) positive (naturally infected) but clinically healthy budgerigars (Melopsittacus undulatus) were inoculated with two isolates of Mycobacterium avium subsp. avium isolated from naturally infected golden pheasant (Chrysolophus pictus) and peafowl (Pavo cristatus). During a period of more than two months after inoculation, samples of cloacal and crop swabs, faeces, and blood were obtained for BFDV and Mycobacterium avium testing with PCR. Birds were euthanized nine weeks after inoculation. All infected budgerigars developed signs typical of mycobacteriosis, but more advanced clinical and pathological changes were visible in the group infected with the pheasant isolate. Only a few cloacal and crop swab samples were positive for Mycobacterium avium subsp. avium despite advanced pathological changes in the internal organs. In the groups infected with mycobacterium isolates the frequency of BFDV-positive samples was higher than in the control group. In the infected groups the frequency of BFDV was substantially higher in the cloacal swabs of birds inoculated with the pheasant isolate than in the peafowl-isolate-infected group.

  15. Mycobacterium avium Infection after Acupoint Embedding Therapy

    Directory of Open Access Journals (Sweden)

    Jiao Zhang, MD

    2017-09-01

    Full Text Available Summary:. Nontuberculous mycobacterium is a ubiquitous environmental organism that is unusual to cause a true infection, but it can cause severe cutaneous infections. In this case report, we present a successful treatment for a Chinese patient with Mycobacterium avium cutaneous infection after acupoint embedding therapy. We managed to conduct pathogenic detection, drug sensitive test, and multidisciplinary consultation. Finally, a systematic treatment strategy of nontuberculous mycobacterium was performed. Twenty-two-month follow-up revealed excellent outcome without any recurrence.

  16. Circumvention of the Mycobactin Requirement of Mycobacterium paratuberculosis

    Science.gov (United States)

    Morrison, Norman E.

    1965-01-01

    Morrison, Norman E. (Johns Hopkins University-Leonard Wood Memorial Leprosy Research Laboratory, Baltimore, Md.). Circumvention of the mycobactin requirement of Mycobacterium paratuberculosis. J. Bacteriol. 89:762–767. 1965.—The mycobactin growth requirement of Mycobacterium paratuberculosis was circumvented on glucose-containing synthetic medium with an initial pH of 5.5. Mycobactin was required during the first transfer on the synthetic medium. Subsequent transfers have grown in the absence of mycobactin. The growth of mycobactin-“independent” strains of M. paratuberculosis on the synthetic medium was found to be stimulated by low concentrations of mycobactin. The circumvention of the mycobactin requirement appears to depend upon the properties of the medium and not upon having created conditions which promote endogenous mycobactin synthesis. Investigation of the glucose-containing synthetic medium showed that: (i) growth stimulatory compounds were formed during autoclaving, and (ii) compared with neutrality a pH of 5.5 gave markedly increased pellicle yields. It was suggested that the growth-stimulatory compounds formed during autoclaving may in part be responsible for the circumvention of the mycobactin requirement. PMID:14273658

  17. 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

    positive. Furthermore, direct MAP contamination of milk was related to infection stages while indirect contamination was associated to within-herd prevalence and distribution of infection stages. Control options implemented included discarding of milk based on diagnostic test results. Median MAP load...... in farm bulk tank milk at within-herd infection prevalences from 7.5 to 60% were estimated to 0.74-6.81 cfu/ml, of which the contribution of direct contamination was less than 1%. Maximum concentration at the prevalence of 60% could be 1186 cfu/ml caused by shedding of high amounts of MAP in feces from...

  18. 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

    and significant bacterial excretion in feces. The proportion of detectable infections among all infected animals that will develop disease is often referred to as ‘the tip of the iceberg’. The purpose of this study was to estimate this proportion. Test-records from 18,972 Danish dairy cows with MAP specific Ig...... 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...

  19. Gamma delta T cells are early responders to Mycobacterium avium subsp. paratuberculosis in colostrum-replete Holstein calves

    Science.gov (United States)

    Peripheral blood mononuclear and mesenteric lymph node cells (PBMC and MNL, respectively) were obtained from 30 calves that were assigned randomly at birth to one of six treatment groups: 1) colostrum deprived (CD), no vitamins; 2) colostrum replacer (CR), no vitamins; 3) CR, vitamin A; 4) CR, vitam...

  20. 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 s...

  1. 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

  2. Assessment of different monoclonal antibodies for the immunohistochemical detection of Mycobacterium avium subsp. paratuberculosis in tissue sections

    OpenAIRE

    Delgado, L.; Colstra, C.; Ferreras, Mª del Carmen; García Marín, Juan Francisco; Bakker, D.; Pérez Pérez, Valentín

    2011-01-01

    1 página.-- Trabajo presentado al II Iberic Meeting of Veterinary Pathology.--XVI Annual Meeting of the Portuguese Society of Animal Pathology.-- Annual Meeting of the Spanisch Society of Veterinary Anatomical Pathology. (Lisboa, Portugal, 1-3 de junio de 2011).

  3. 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 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....

  4. 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

    consisting of undefined antigens with possible cross reactions toward other environmental bacteria. The objective of the study was to optimize the IFN-γ test using different types of novel antigens for stimulation. Fourteen novel antigen candidates were selected for testing, including 4 peptides of the ESAT...... on the same 30 heifers from a known MAP infected herd. Determination of cut-off for each antigen was based on samples from a non-infected herd, including 60 heifers. Based on PPDj stimulations, more than 50% of the heifers tested MAP positive at the first two samplings, whereas only 20% tested positive...... at third sampling. The resulted showed that PPDj detect a high percentage as MAP positive animals, as this crude antigen mixture is expected to induce non-specific IFN-γ production. However, the tested latency antigens, some secreted proteins and some peptides of the ESAT-6 family detected a comparable...

  5. Characterization of a Mycobacterium avium subsp. avium Operon Associated with Virulence and Drug Detoxification

    Directory of Open Access Journals (Sweden)

    Mariana Noelia Viale

    2014-01-01

    Full Text Available The lprG-p55 operon of Mycobacterium tuberculosis and Mycobacterium bovis is involved in the transport of toxic compounds. P55 is an efflux pump that provides resistance to several drugs, while LprG is a lipoprotein that modulates the host's immune response against mycobacteria. The knockout mutation of this operon severely reduces the replication of both mycobacterial species during infection in mice and increases susceptibility to toxic compounds. In order to gain insight into the function of LprG in the Mycobacterium avium complex, in this study, we assayed the effect of the deletion of lprG gene in the D4ER strain of Mycobacterium avium subsp. avium. The replacement of lprG gene with a hygromycin cassette caused a polar effect on the expression of p55. Also, a twofold decrease in ethidium bromide susceptibility was observed and the resistance to the antibiotics rifampicin, amikacin, linezolid, and rifabutin was impaired in the mutant strain. In addition, the mutation decreased the virulence of the bacteria in macrophages in vitro and in a mice model in vivo. These findings clearly indicate that functional LprG and P55 are necessary for the correct transport of toxic compounds and for the survival of MAA in vitro and in vivo.

  6. Labdane Diterpenoids from Marrubium globosum ssp. libanoticum.

    Science.gov (United States)

    Rigano, Daniela; Grassia, Armando; Bruno, Maurizio; Rosselli, Sergio; Piozzi, Franco; Formisano, Carmen; Arnold, Nelly Apostolides; Senatore, Felice

    2006-05-01

    From the aerial parts of Marrubium globosum ssp. libanoticum, seven labdane diterpenoids were isolated. Three of them are new natural products [(13R)-9alpha,13alpha-epoxylabda-6beta(19),16(15)-diol dilactone (2), deacetylvitexilactone (7), marrulanic acid (8)], whereas the other four, namely, (13S)-9alpha,13alpha-epoxylabda-6beta(19),16(15)-diol dilactone (1), cyllenin A (3), 15-epi-cyllenin A (4), and marrulibanoside, are previously known compounds. The structures of 2, 7, and 8 were determined by spectroscopic and chemical methods.

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

    Directory of Open Access Journals (Sweden)

    K Parvandar Asadollahi

    2015-01-01

    In conclusion: It is suggested that more DNA fingerprinting tests for non-tuberculous Mycobacteria, particularly M. avium complex isolated from infected birds and humans, be conducted to find the source of their infections.

  8. GENETIC FINGERPRINTING OF MYCOBACTERIUM AVIUM COMPLEX (MAC) ORGANISMS ISOLATED FROM HOSPITAL PATIENTS AND THE ENVIRONMENT

    Science.gov (United States)

    A particularly pathogenic group of mycobacteria belong to the Mycobacterium avium complex (MAC), which includes M. avium and M. intracellulare. MAC organisms cause disease in children, the elderly, and immuno-compromised individuals. A critical step in preventing MAC infections...

  9. Generation and characterization of Brassica rapa ssp. pekinensis ...

    Indian Academy of Sciences (India)

    2015-09-11

    Sep 11, 2015 ... Abstract. Five monosomic alien addition lines (MAALs) of Brassica rapa ssp. pekinensis – B. oleracea var. capitata were obtained by hybridization and backcrossing between B. rapa ssp. pekinensis (female parent) and B. oleracea var. capitata. The alien link- age groups were identified using 42 B. oleracea ...

  10. Complete genome sequence of the hippuricase-positive Campylobacter avium type strain LMG 24591

    Science.gov (United States)

    Campylobacter avium is a hippurate-positive, thermotolerant campylobacter that has been isolated from poultry. Here we present the genome sequences of two C. avium strains isolated from broiler chickens: strains LMG 24591T (complete genome) and LMG 24592 (draft genome). The C. avium type strain geno...

  11. 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

  12. 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.

  13. High active potential antimycobacterial agents against Mycobacterium avium.

    Science.gov (United States)

    Karel, Waisser; Jiří, Kuneš; Jiřina, Stolaříková

    2012-06-01

    Derivatives of 3-phenyl-2H-1,3-benzoxazine-2,4(3H)-dione are active against Mycobacterium avium when substituted in position 7 with a methyl. One or two carbonyl groups have to be replaced with a thioxo group. High active derivatives are the compounds without substitution on the phenyl, or those substituted on the phenyl in position 3 or 4 with chlorine, bromine or a methyl. The substitution in position 3 and 4 with two atoms of chlorine lowers the activity. The compounds are active against INH resistant strains. We synthesized other 44 derivatives with a similar structure of the compounds as in the paper but substituted in position 7 with other substituents (chlorine, bromine methoxy). The activity against M. avium was poor. It can be concluded that a new group of compounds with an excellent activity against M. avium has been found.

  14. Isolation, Identification, and Characterization of a New Highly Pathogenic Field Isolate of Mycobacterium avium spp. avium

    Directory of Open Access Journals (Sweden)

    Liangquan Zhu

    2018-01-01

    Full Text Available Avian tuberculosis is a chronic, contagious zoonotic disease affecting birds, mammals, and humans. The disease is most often caused by Mycobacterium avium spp. avium (MAA. Strain resources are important for research on avian tuberculosis and vaccine development. However, there has been little reported about the newly identified MAA strain in recent years in China. In this study, a new strain was isolated from a fowl with symptoms of avian tuberculosis by bacterial culture. The isolated strain was identified to be MAA by culture, staining, and biochemical and genetic analysis, except for different colony morphology. The isolated strain was Ziehl-Zeelsen staining positive, resistant to p-nitrobenzoic acid, and negative for niacin production, Tween-80 hydrolysis, heat stable catalase and nitrate production. The strain had the DnaJ gene, IS1245, and IS901, as well. Serum agglutination indicated that the MAA strain was of serotype 1. The MAA strain showed strong virulence via mortality in rabbits and chickens. The prepared tuberculin of the MAA strain had similar potency compared to the MAA reference strain and standard tuberculin via a tuberculin skin test. Our studies suggested that this MAA strain tends to be a novel subtype, which might enrich the strain resource of avian tuberculosis.

  15. School Teams up for SSP Functional Models

    Science.gov (United States)

    Pignolet, G.; Lallemand, R.; Celeste, A.; von Muldau, H.

    2002-01-01

    Space Solar Power systems appear increasingly as one of the major solutions to the upcoming global energy crisis, by collecting solar energy in space where this is most easy, and sending it by microwave beam to the surface of the planet, where the need for controlled energy is located. While fully operational systems are still decades away, the need for major development efforts is with us now. Yet, for many decision-makers and for most of the public, SSP often still sounds like science fiction. Six functional demonstration systems, based on the Japanese SPS-2000 concept, have been built as a result of a cooperation between France and Japan, and they are currently used extensively, in Japan, in Europe and in North America, for executive presentations as well as for public exhibitions. There is demand for more models, both for science museums and for use by energy dedicated groups, and a senior high school in La Reunion, France, has picked up the challenge to make the production of such models an integrated practical school project for pre-college students. In December 2001, the administration and the teachers of the school have evaluated the feasibility of the project and eventually taken the go decision for the school year 2002- 2003, when for education purposes a temporary "school business company" will be incorporated with the goal to study and manufacture a limited series of professional quality SSP demonstration models, and to sell them world- wide to institutions and advocacy groups concerned with energy problems and with the environment. The different sections of the school will act as the different services of an integrated business : based on the current existing models, the electronic section will redesign the energy management system and the microwave projector module, while the mechanical section of the school will adapt and re-conceive the whole packaging of the demonstrator. The French and foreign language sections will write up a technical manual for

  16. Etude chimique et biologique des huiles essentielles de Juniperus phoenicea ssp. lycia et Juniperus phoenicea ssp. turbinata du Maroc

    Directory of Open Access Journals (Sweden)

    Mansouri, N.

    2011-01-01

    Full Text Available Chemical and biological study of essential oils of Moroccan Juniperus phoenicea ssp. lycia and Juniperus phoenicea ssp. turbinata. The composition of the essential oils of the branches and berries of Juniperus phoenicea (Cupressaceae, J. phoenicea ssp. lycia (plain and J. phoenicea ssp. turbinata (mountain, obtained by hydrodistillation, collected from Morocco, was analyzed by GC and GC/MS. The yields of essential oils were varying in function of the subspecies and of the part of the plant studied. The essential oils of these tree species are largely dominated by α-pinene and may be an important source of this component of a great interest on the international market. The effectiveness of essential oils from branches of the subspecies lycia against fungi decay wood has also been emphasized.

  17. Involvement of Stat1 in the Phagocytosis of M. avium

    Directory of Open Access Journals (Sweden)

    Sabrina Dominici

    2012-01-01

    Full Text Available Mycobacterium avium is an intracellular pathogen preferentially infecting human macrophages where they activate the JAK/STAT1 pathway. This activation enhances the survival of infected cells, but, at the same time, makes macrophages optimal targets for drugs development against p-tyr701stat1. In this study, we demonstrate that the fast and transient activity of the JAK/STAT1 pathway occurs immediately after macrophages internalization of heat-killed M. avium or inert particles. Furthermore, we show that a persistent Stat1 pathway activation occurs only when an intracellular M. avium infection is established in macrophages. These results strongly indicate different mechanisms of p-tyr701Stat1 activation. In particular, here we report findings aiming at explaining the short-time enhancement of p-tyr701Stat1 and shows its predominant relationship with FcγRs engagement during the internalization process. Furthermore, we demonstrate that opsonized live M. avium is phagocytosed by macrophages involving membrane receptors not related with JAK/STAT1 signalling pathway. On the contrary, heat-inactivated bacilli or latex particles seem to be internalized only after involvement of FcγRs and subsequent Stat1 phosphorylation.

  18. Cryptosporidium avium n. sp (Apicomplexa: Cryptosporidiidae) in birds

    Czech Academy of Sciences Publication Activity Database

    Holubová, Nikola; Sak, Bohumil; Horčičková, Michaela; Hlásková, Lenka; Květoňová, Dana; Menchaca, S.; McEvoy, J.; Kváč, Martin

    2016-01-01

    Roč. 115, č. 6 (2016), s. 2243-2251 ISSN 0932-0113 R&D Projects: GA ČR GA15-01090S Institutional support: RVO:60077344 Keywords : Cryptosporidium avium * morphology * molecular analyses * transmission studies * Cryptosporidium avian genotype V Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.329, year: 2016

  19. Characterization and expression of secA in Mycobacterium avium.

    Science.gov (United States)

    Limia, A; Sangari, F J; Wagner, D; Bermudez, L E

    2001-04-13

    Mycobacterium avium is both a pathogen that infects several hosts such as humans, pigs, and birds, as well as a microorganism that is encountered in environmental sources (soil and water). Protein secretion by the bacterium is likely to influence its ability to overcome adverse and competitive conditions both within or outside the host. Using a combination of cloning and information available in the databank, we characterized the secA gene from M. avium, encoding for a major preprotein translocase subunit associated with the secretion system of prokaryotics. In addition, we cloned the secA promoter sequence in a reporter construct upstream of a promoterless gfp. It was determined that the secA of M. avium shares large homology with the secA of Mycobacterium tuberculosis but not with secA of Mycobacterium leprae. secA expression was determined to be greater at logarithmic growth phase although it was also expressed at low levels during the stationary phase. secA expression was also observed when the bacteria were incubated in water as well as within human monocyte-derived macrophages and in conditions that are associated with biofilm formation. Future evaluation of the sec pathway in M. avium might provide important information about secreted proteins that are required for survival in different environments.

  20. The SSP4: A world of deepening inequality

    Energy Technology Data Exchange (ETDEWEB)

    Calvin, Katherine; Bond-Lamberty, Ben; Clarke, Leon; Edmonds, James; Eom, Jiyong; Hartin, Corinne; Kim, Sonny; Kyle, Page; Link, Robert; Moss, Richard; McJeon, Haewon; Patel, Pralit; Smith, Steve; Waldhoff, Stephanie; Wise, Marshall

    2017-01-01

    The Shared Socioeconomic Pathway 4 (SSP4), “Inequality” or “A Road Divided,” is one of the five SSPs developed to guide the creation of new scenarios for the “Parallel Process”. We describe, in quantitative terms, the SSP4 as implemented by the Global Change Assessment Model (GCAM), the marker model for this scenario. We use demographic and economic assumptions, in combination with technology and non-climate policy assumptions to develop a quantitative representation of energy, land-use and land-cover that are consistent with the SSP4 storyline. The resulting scenario is one with stark differences across regions. High-income regions prosper, continuing to increase their demand for energy and food. Electrification increases in these regions, with the increased generation being met by nuclear and renewables. Low-income regions, however, stagnate due to limited growth in income. These regions continue to depend on traditional biofuels, leading to high pollutant emissions. Due to a declining dependence on fossil fuels in all regions, total radiative forcing only reaches 6.4 Wm-2 in 2100, making this a world with relatively low challenges to mitigation. We explore the effects of mitigation effort on the SSP4 world, finding that the imposition of a carbon price has a varied effect across regions. In particular, the SSP4 mitigation scenarios are characterized by afforestation in the high-income regions and deforestation in the low-income regions. Finally, we compare the GCAM SSP4 results to other integrated assessment model (IAM) quantifications of the SSP4 and to other SSPs, both those generated by GCAM and those of the other IAMs.

  1. In-Silico identification of peptides for the diagnostics of paratuberculosis

    DEFF Research Database (Denmark)

    Tang, Sheila Tuyet; Lund, Ole; Jungersen, Gregers

    Identification of bovine MHC class II reactive peptides that are specific/unique to paratuberculosis and conserved across pathogenic variations of the paratuberculosis proteome will be of high value for development of new vaccines and immune based diagnostics. Here, we present an in silico...... consecutive amino acids was found. This resulted in approximately 80000 20mers that were 100% conserved between the two positive strains, sharing no 8mer overlap to any negative genome. Residual 20mers were next in-silico checked for binding to each of five prevalent bovine class-II MHC molecules using...

  2. Isolation and identification of Mycobacterium avium subspecies silvaticum from a horse.

    Science.gov (United States)

    Chiers, Koen; Deschaght, Pieter; De Baere, Thierry; Dabrowski, Slawomir; Kotlowski, Roman; De Clercq, Dominique; Ducatelle, Richard; Vaneechoutte, Mario

    2012-07-01

    Routine cultivation methods are able to distinguish between isolates of the Mycobacterium avium and the Mycobacterium tuberculosis complex. However, molecular tools are needed to further identify the several subspecies in the M. avium complex, especially for the subspecies avium and silvaticum. A rapid technique using HhaI restriction digestion of a 349 bp amplification product of the 85B antigen (α-antigen) gene was used for the identification of M. avium subsp. silvaticum in a three-year-old gelding presenting with caseous, necrotizing, granulomatous lesions. The result was confirmed by sequencing of the 85B antigen gene. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. Risk analysis of Safety Service Patrol (SSP) systems in Virginia.

    Science.gov (United States)

    Dickey, Brett D; Santos, Joost R

    2011-12-01

    The transportation infrastructure is a vital backbone of any regional economy as it supports workforce mobility, tourism, and a host of socioeconomic activities. In this article, we specifically examine the incident management function of the transportation infrastructure. In many metropolitan regions, incident management is handled primarily by safety service patrols (SSPs), which monitor and resolve roadway incidents. In Virginia, SSP allocation across highway networks is based typically on average vehicle speeds and incident volumes. This article implements a probabilistic network model that partitions "business as usual" traffic flow with extreme-event scenarios. Results of simulated network scenarios reveal that flexible SSP configurations can improve incident resolution times relative to predetermined SSP assignments. © 2011 Society for Risk Analysis.

  4. Vibrio vulnificus bacteriophage SSP002 as a possible biocontrol agent.

    Science.gov (United States)

    Lee, Hyun Sung; Choi, Slae; Shin, Hakdong; Lee, Ju-Hoon; Choi, Sang Ho

    2014-01-01

    A novel Vibrio vulnificus-infecting bacteriophage, SSP002, belonging to the Siphoviridae family, was isolated from the coastal area of the Yellow Sea of South Korea. Host range analysis revealed that the growth inhibition of phage SSP002 is relatively specific to V. vulnificus strains from both clinical and environmental samples. In addition, a one-step growth curve analysis and a bacteriophage stability test revealed a latent period of 65 min, a burst size of 23 ± 2 PFU, as well as broad temperature (20°C to 60°C) and pH stability (pH 3 to 12) ranges. A Tn5 random transposon mutation of V. vulnificus and partial DNA sequencing of the inserted Tn5 regions revealed that the flhA, flhB, fliF, and fleQ mutants are resistant to SSP002 phage infection, suggesting that the flagellum may be the host receptor for infection. The subsequent construction of specific gene-inactivated mutants (flhA, flhB, fliF, and fleQ) and complementation experiments substantiated this. Previously, the genome of phage SSP002 was completely sequenced and analyzed. Comparative genomic analysis of phage SSP002 and Vibrio parahaemolyticus phage vB_VpaS_MAR10 showed differences among their tail-related genes, supporting different host ranges at the species level, even though their genome sequences are highly similar. An additional mouse survival test showed that the administration of phage SSP002 at a multiplicity of infection of 1,000 significantly protects mice from infection by V. vulnificus for up to 2 months, suggesting that this phage may be a good candidate for the development of biocontrol agents against V. vulnificus infection.

  5. Prevalence of paratuberculosis in the dairy goat and dairy sheep industries in Ontario, Canada

    DEFF Research Database (Denmark)

    Bauman, Cathy A.; Jones-Bitton, Andria; Menzies, Paula

    2016-01-01

    A cross-sectional study was undertaken (October 2010 to August 2011) to estimate the prevalence of paratuberculosis in the small ruminant dairy industries in Ontario, Canada. Blood and feces were sampled from 580 goats and 397 sheep (lactating and 2 y of age or older) that were randomly selected...

  6. Mycobacterium avium-intracellulare: a rare cause of subacromial bursitis.

    Science.gov (United States)

    Sinha, Raj; Tuckett, John; Hide, Geoff; Dildey, Petra; Karsandas, Alvin

    2015-01-01

    Septic subacromial bursitis is an uncommon disorder with only a few reported cases in the literature. The most common causative organism is Staphylococcus aureus. We report the case of a 61-year-old female with a septic subacromial bursitis where the causative organism was found to be Mycobacterium avium-intracellulare (MAI). The diagnosis was only made following a biopsy, and we use this case to highlight the importance of recognising the need to consider a biopsy and aspiration in atypical situations.

  7. Determination of chromosomal ploidy in Agave ssp. | Lingling ...

    African Journals Online (AJOL)

    Chromosome observation is necessary to elucidate the structure, function and organization of Agave plants' genes and genomes. However, few researches about chromosome observation of Agave ssp. were done, not only because their chromosome numbers are large, but also because their ploidies are complicated.

  8. Determination of chromosomal ploidy in Agave ssp. | Lingling ...

    African Journals Online (AJOL)

    The root tips of 19 Agave ssp. germplasms were used as materials for determining their chromosomal ploidies. Through normal pre-treatment, fixation, digesting and Giemsa staining, the glass slides with expelled cells on them were obtained. Observed with a light microscope, the results showed that 10 germplasms are ...

  9. Campylobacter fetus ssp jejuni en patología humana

    Directory of Open Access Journals (Sweden)

    Luz Echeverri

    1982-06-01

    Full Text Available Algunos microorganismos, como los Rotovirus y el Compylobocter fetus ssp jejuni reconocidos y estudiados recientemente, han llegado a ocupar un lugar preponderante en el grupo de enteropatógenos considerados como problema en salud pública.

  10. Antihypertensive effect of extracts from Crateva adansonii DC.ssp ...

    African Journals Online (AJOL)

    Crateva adansonii DC.ssp. adansonii (CA) is a medicinal plant used in traditional medicine to treat various diseases including hypertension. The main objective of this study was to assess the effect of the plant on high blood pressure. The crude aqueous extract of CA has undergone a liquid-liquid fractionation with ...

  11. 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...

  12. Detection of quantification of Mycobacterium avium complex organisms in drinking water

    Science.gov (United States)

    The Mycobacterium avium Complex (MAC) includes the species M. avium (MA), M. intracellulare (MI), and others. MAC are listed on the U. S. Environmental Protection Agency’s Contaminant Candidate List 2 (CCL2) due to their association with human disease and occurrence in public dr...

  13. REAL-TIME QUANTITATIVE PCR DETECTION OF MYCOBACTERIUM AVIUM COMPLEX ORGANISMS IN DRINKING WATER

    Science.gov (United States)

    The Mycobacterium avium Complex (MAC) includes the species M. avium (MA), M. intracellulare (MI), and others. MAC are listed on the U.S. Environmental Protection Agency's Contaminant Candidate List (CCL) due to their association with human disease and occurrence in public drinkin...

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

    Science.gov (United States)

    Members of the Mycobacterium avium complex (MAC) are naturally occurring bacteria in the environment. A link has been suggested between M. avium strains in drinking water and clinical isolates from infected individuals. There is a need to develop new screening methodologies tha...

  15. Isolation of mycobacteria other than Mycobacterium avium from porcine lymph nodes

    NARCIS (Netherlands)

    Ingen, van J.; Wisselink, H.J.; Solt-Smits, van C.B.; Boeree, M.J.; Soolingen, D.

    2010-01-01

    Mycobacterium avium causes lymphadenitis in pigs. This presents an economical burden, as these pigs meat is considered inappropriate for consumption. In humans, lymphadenitis due to nontuberculous mycobacteria (NTM) primarily affects children and is caused by a variety of NTM, though M. avium

  16. Isolation of mycobacteria other than Mycobacterium avium from porcine lymph nodes.

    NARCIS (Netherlands)

    Ingen, J. van; Wisselink, H.J.; Solt-Smits, C.B. van; Boeree, M.J.; Soolingen, D. van

    2010-01-01

    Mycobacterium avium causes lymphadenitis in pigs. This presents an economical burden, as these pigs meat is considered inappropriate for consumption. In humans, lymphadenitis due to nontuberculous mycobacteria (NTM) primarily affects children and is caused by a variety of NTM, though M. avium

  17. 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.

  18. Chemical and Antibacterial Polymorphism of Juniperus oxycedrus ssp. oxycedrus and Juniperus oxycedrus ssp. macrocarpa (Cupressaceae Leaf Essential Oils from Tunisia

    Directory of Open Access Journals (Sweden)

    Hnène Medini

    2013-01-01

    Full Text Available Essential oils from Juniperus oxycedrus L. have been used since antiquity for fragrance, flavoring, medicinal, antimicrobial, insecticidal, and cosmetic purposes. Several works studied the chemical composition of the essential oils of Juniperus oxycedrus leaves. The aim of this study is to investigate the chemotaxonomic relationships and antibacterial activity of two Tunisian subspecies: Juniperus oxycedrus ssp. oxycedrus (L. K. Deb. and Juniperus oxycedrus ssp. macrocarpa (S. & m. Ball. In addition, and for the first time, we reported the antibacterial activities of Tunisian J. oxycedrus ssp. macrocarpa and J. oxycedrus ssp. oxycedrus against four bacteria. Essential oils obtained by hydrodistillation were analysed by GC and GC/MS. Fifty-five constituents were identified. Thirty four major compounds were retained for the study of the chemical variability, and α-pinene, sylvestrene, p-cymene, and 13-epi-manoyl oxide were the main ones. The chemical principal components analysis (PCA identified three chemotypes. The study of the antibacterial activity showed that Escherichia coli was found to be extremely resistant (zone diameter 0 mm to all the oils tested, while Staphylococcus aureus was the most sensitive strain (zone diameter 13.5 mm and MIC ranged from 600 to 650 μg/mL.

  19. Effect of pasteurization on survival of Mycobacterium paratuberculosis in milk.

    Science.gov (United States)

    Gao, A; Mutharia, L; Chen, S; Rahn, K; Odumeru, J

    2002-12-01

    Mycobacterium paratuberculosis (Mptb) is the causative agent of Johne's disease of ruminant animals including cattle, goats, and sheep. It has been suggested that this organism is associated with Crohn's disease in humans, and milk is a potential source of human exposure to this organism. A total of 18, including 7 regular batch and 11 high temperature short time (HTST) pasteurization experiments, were conducted in this study. Raw milk or ultra-high temperature pasteurized milk samples were spiked at levels of 10(3), 10(5), and 10(7) cfu of Mptb/ml. Escherichia coli and Mycobacterium bovis BCG strains at 10(7) cfu/ml were used as controls. Pasteurization experiments were conducted using time and temperature standards specified in the Canadian National Dairy Code: regular batch pasteurization method: 63 degrees C for 30 min, and HTST method: 72 degrees C for 15 s. The death curve of this organism was assessed at 63 degrees C. No survivors were detected after 15 min. Each spiked sample was cultured in Middlebrook 7H9 culture broth and Middlebrook 7H11 agar slants. Samples selected from 15 experiments were also subjected to BACTEC culture procedure. Survival of Mptb was confirmed by IS900-based PCR of colonies recovered on slants. No survivors were detected from any of the slants or broths corresponding to the seven regular batch pasteurization trials. Mptb survivors were detected in two of the 11 HTST experiments. One was by both slant and broth culture for the sample spiked to 10(7) cfu/ml of Mptb, while the other was detected by BACTEC for the sample spiked to 10(5) cfu/ml. These results indicate that Mptb may survive HTST pasteurization when present at > or = 10(5) cfu/ml in milk. A total of 710 retail milk samples collected from retail store and dairy plants in southwest Ontario were tested by nested IS900 PCR for the presence of Mptb. Fifteen percent of these samples (n = 110) were positive. However, no survivors were isolated from the broth and agar cultures of

  20. Competition for Antigen between Th1 and Th2 Responses Determines the Timing of the Immune Response Switch during Mycobaterium avium Subspecies paratuberulosis Infection in Ruminants

    Science.gov (United States)

    Magombedze, Gesham; Eda, Shigetoshi; Ganusov, Vitaly V.

    2014-01-01

    Johne's disease (JD), a persistent and slow progressing infection of ruminants such as cows and sheep, is caused by slow replicating bacilli Mycobacterium avium subspecies paratuberculosis (MAP) infecting macrophages in the gut. Infected animals initially mount a cell-mediated CD4 T cell response against MAP which is characterized by the production of interferon (Th1 response). Over time, Th1 response diminishes in most animals and antibody response to MAP antigens becomes dominant (Th2 response). The switch from Th1 to Th2 response occurs concomitantly with disease progression and shedding of the bacteria in feces. Mechanisms controlling this Th1/Th2 switch remain poorly understood. Because Th1 and Th2 responses are known to cross-inhibit each other, it is unclear why initially strong Th1 response is lost over time. Using a novel mathematical model of the immune response to MAP infection we show that the ability of extracellular bacteria to persist outside of macrophages naturally leads to switch of the cellular response to antibody production. Several additional mechanisms may also contribute to the timing of the Th1/Th2 switch including the rate of proliferation of Th1/Th2 responses at the site of infection, efficiency at which immune responses cross-inhibit each other, and the rate at which Th1 response becomes exhausted over time. Our basic model reasonably well explains four different kinetic patterns of the Th1/Th2 responses in MAP-infected sheep by variability in the initial bacterial dose and the efficiency of the MAP-specific T cell responses. Taken together, our novel mathematical model identifies factors of bacterial and host origin that drive kinetics of the immune response to MAP and provides the basis for testing the impact of vaccination or early treatment on the duration of infection. PMID:24415928

  1. 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

    option. Median MAP load in farm bulk tank milk at within-herd infection prevalences from 7.5% to 60% were estimated to be 0.54-7.53 CFU/mL milk. Maximum concentration at the prevalence of 60% could be 1186 CFU/mL caused by shedding of high amounts of MAP in feces. At the prevalence of 15%, discarding...

  2. Cellular responses to Mycobacterium avium, subsp. paratuberculosis in colostrum-deprived and colostrum-replete holstein calves supplemented with fat-soluble vitamins

    Science.gov (United States)

    Immune benefits of colostrum are attributed to passively transferred IgG but also to growth factors, cytokines, antimicrobial peptides, and leukocytes. Non-nutritive compounds in colostrum promote Th2-biased immune responses to early microbial encounters and prevent harmful, inappropriate inflammat...

  3. Detection of Mycobacterium avium subsp. Paratuberculosis in Cattle by using Indirect Absorbed ELISA (enzyme-linked immunosorbent assay system and culture in Alborz Province, Iran

    Directory of Open Access Journals (Sweden)

    Hedyeh Teymouri

    2016-01-01

    Conclusion: Because there is no treatment or cure for Johne's disease, detection of infected cattle and subsequent culling is very important for preventing infection in other cattle. Fecal culture is a standard method for the identification of the disease. However, in Johne's disease, due to prolonged incubation and shedding of the disease, the probability of isolating the responsible agent is very low. To identify the infection, the indirect absorbed ELISA method is used for eradication. This technique is considered as one of the most reliable for identification of the disease worldwide due to its ease of use and low cost. However, for confirmation of ELISA-positive results, culture method has been recommended.

  4. 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 population...... for 24h revealed the presence of a subpopulation of cells probably resistant to the antimicrobial compounds tested. Use of nisin and bacteriocin-producing LAB strains could lead to new intervention strategies for the control of MAP based on in vivo application of probiotic cultures as feed additives...

  5. Within- and between-herd prevalence variation of Mycobacterium avium subsp. paratuberculosis infection among control programme herds in Denmark (2011-2013)

    DEFF Research Database (Denmark)

    Verdugo, Cristobal; Toft, Nils; Nielsen, Søren

    2015-01-01

    estimates. Bayesian posterior probabilities were computed in order to compare prevalence between the years. A total of 665,700 samples were included in the study, from 221,914, 224,040, and 220,466 cows sourced from 1138, 1112, and 1059 herds in years 2011, 2012, and 2013, respectively. In that period, HTP...... estimates of 0.92 (95% posterior probability interval (PPI), 0.87-0.96), 0.78 (95% PPI, 0.74-0.83), and 0.75 (95% PPI, 0.71-0.78) were recorded, respectively. Low TP were observed, with population mean estimates of 0.08 (95% PPI, 0.07-0.08), 0.07 (95% PPI, 0.07-0.08), and 0.07 (95% PPI, 0.......06-0.07) for the three consecutive years. Statistically-important differences were recorded for HTP and population mean TP estimates between years, indicating a trend for a decreasing level of MAP infection at both herd and animal level. Model results showed that MAP infection was widespread among the Dairy cattle herds...

  6. 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...

  7. Chemical characteristic of salt fermented meat inoculated with Pediococcus ssp

    Science.gov (United States)

    Pramono, Y. B.; Rahayu, E. S.; Suparmo; Utami, T.; Nurwantoro; Yunianto, V. D.

    2018-01-01

    The research goal is knowing of the characteristict of salt fermented meat by Pediococcus ssp. There were microbiological, chemical, and off-flavor compound during fermentation. This study was conducted on research of influence of salt-meat fermentation inoculated used starter. They were included microbiological characteristics, and chemical characteristics. Microbiological characteristics observed were total bacteria, number of coliform groups, bacteria producing bioamine, and total lactic acid bacteria. The result showed that decreasing of coliform and bioamine producer bacteria, and total lactic acid bacteria decreased 3 log cycle. While the soluble protein increased of 7-8% and bioamine increased of 5-6 mg/100 g. And then Off-flavour compound, TVN and TMA increased of 36-20 mg/100g and 16-30 mg/100g, respectively. Conclusion of the research that Pedioccoccus ssp. influenced salt fermented meat.

  8. Lactococcus lactis ssp. lactis as Potential Functional Starter Culture

    Directory of Open Access Journals (Sweden)

    Jelena Cvrtila

    2014-01-01

    Full Text Available The aim of this study is to identify and characterise potential autochthonous functional starter cultures in homemade horsemeat sausage. The dominant microflora in the samples of horsemeat sausage were lactic acid bacteria (LAB, followed by micrococci. Among the LAB, Lactococcus lactis ssp. lactis and Lactobacillus plantarum were the dominant species, and since the first is not common in fermented sausages, we characterised it as a potential functional starter culture. Lactococcus lactis ssp. lactis produced a significant amount of lactic acid, displayed good growth capability at 12, 18 and 22 °C, growth in the presence of 5 % NaCl, good viability after lyophilisation and in simulated gastric and small intestinal juice, antimicrobial activity against test pathogens, and good adhesive properties in vitro.

  9. Lactococcus lactis ssp. lactis as Potential Functional
Starter Culture

    Science.gov (United States)

    Cvrtila, Jelena; Topić, Ivana; Delaš, Frane; Markov, Ksenija

    2014-01-01

    Summary The aim of this study is to identify and characterise potential autochthonous functional starter cultures in homemade horsemeat sausage. The dominant microflora in the samples of horsemeat sausage were lactic acid bacteria (LAB), followed by micrococci. Among the LAB, Lactococcus lactis ssp. lactis and Lactobacillus plantarum were the dominant species, and since the first is not common in fermented sausages, we characterised it as a potential functional starter culture. Lactococcus lactis ssp. lactis produced a significant amount of lactic acid, displayed good growth capability at 12, 18 and 22 °C, growth in the presence of 5% NaCl, good viability after lyophilisation and in simulated gastric and small intestinal juice, antimicrobial activity against test pathogens, and good adhesive properties in vitro. PMID:27904322

  10. Estudio etnobotánico, arquitectura foliar y anatomía vegetativa de Agastache mexicana ssp. mexicana y A. mexicana ssp. xolocotziana Etnobotany, leaf architecture, and vegetative anatomy of Agastache mexicana ssp. mexicana and A. mexicana ssp. xolocotziana

    Directory of Open Access Journals (Sweden)

    Magali A. Santillán-Ramírez

    2008-12-01

    Full Text Available El presente trabajo incluye información etnobotánica, así como la arquitectura foliar y anatomía vegetativa de Agastache mexicana ssp. mexicana (toronjil morado y A. mexicana ssp. xolocotziana (toronjil blanco, con la finalidad de que sean incluidas en la Farmacopea Herbolaria de los Estados Unidos Mexicanos (FHEUM y que sus caracteres anatómicos se utilicen para validar la identidad del material vegetal en el control de calidad. El registro de la información etnobotánica se efectuó por medio de entrevistas semiestructuradas a pobladores del municipio de Temoaya, Estado de México, México. Para el estudio anatómico y arquitectura foliar, se elaboraron preparaciones semipermanentes y permanentes de tallos y hojas. Ambos toronjiles son utilizados por los habitantes de la zona de estudio como recurso terapéutico y de ornato; los distinguen por la forma de las hojas, el color de la flor y el aroma, pero no existen preferencias de uso entre ambos. La arquitectura foliar sigue el patrón acródromo de posición basal con un desarrollo imperfecto; las areolas muestran un desarrollo imperfecto con disposición al azar, con vénulas simples o ramificadas una vez y el margen con dientes cunonioides en ambas especies. Diferencias anatómicas se observan en el tallo y son los tricomas no glandulares uniseriados en mayor abundancia y las bandas de esclerénquima más desarrolladas los caracteres que distinguen a A. mexicana ssp. xolocotziana de A. mexicana ssp. mexicana.This work provides information on ethnobotany, leaf architecture and anatomy of Agastache mexicana ssp. mexicana (toronjil morado and A. mexicana ssp. xolocotziana (toronjil blanco, with the purpose that they will be included in the Pharmacopeia Herbolaria of the Estados Unidos Mexicanos (FHEUM and that their anatomical characters could be used to validate the identity of the botanical material in quality control. We gathered ethnobotanical information using semi

  11. Mycobacterium avium genotype is associated with the therapeutic response to lung infection

    Science.gov (United States)

    Kikuchi, T; Kobashi, Y; Hirano, T; Tode, N; Santoso, A; Tamada, T; Fujimura, S; Mitsuhashi, Y; Honda, Y; Nukiwa, T; Kaku, M; Watanabe, A; Ichinose, M; Drancourt, M

    2014-01-01

    Factors that can interfere with the successful treatment of Mycobacterium avium lung infection have been inadequately studied. To identify a potent predictor of therapeutic responses of M. avium lung infection, we analyzed variable number tandem repeats (VNTR) at 16 minisatellite loci of M. avium clinical isolates. Associations between the VNTR profiling data and a therapeutic response were evaluated in 59 subjects with M. avium lung infection. M. avium lung infection of 30 subjects in whom clarithromycin-containing regimens produced microbiological and radiographic improvement was defined as responsive disease, while that of the remaining 29 subjects was defined as refractory disease. In phylogenetic analysis using the genotypic distance aggregated from 16-dimensional VNTR data, 59 M. avium isolates were divided into three clusters, which showed a nearly significant association with therapeutic responses (p 0.06). We then subjected the raw 16-dimensional VNTR data directly to principal component analysis, and identified the genetic features that were significantly associated with the therapeutic response (p VNTR data from only four minisatellite loci. In conclusion, we identified four mycobacterial minisatellite loci that together were associated with the therapeutic response of M. avium lung infections. PMID:23829301

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

    DEFF Research Database (Denmark)

    Askgaard, D. S.; Giese, Steen Bjørck; Thybo, S.

    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....

  13. Mycobacterium avium complex disseminated infection in a kidney transplant recipient.

    Science.gov (United States)

    Fadlallah, J; Rammaert, B; Laurent, S; Lanternier, F; Pol, S; Franck, N; Mamzer, M F; Dupin, N; Lortholary, O

    2016-02-01

    Mycobacterium avium-intracellulare complex (MAC) infections are well known in immunocompromised patients, notably in human immunodeficiency virus infection, but remain scarcely described in kidney transplantation. Moreover, cutaneous involvement in this infection is very unusual. We describe here a disseminated infection caused by MAC in a kidney transplant recipient revealed by cutaneous lesions. This case highlights the need for an exhaustive, iterative microbiologic workup in the context of an atypical disease presentation in a renal transplant patient, regardless of the degree of immunosuppression. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. Assessment of listing and categorisation of animal diseases within the framework of the Animal Health Law (Regulation (EU) No 2016/429): paratuberculosis

    DEFF Research Database (Denmark)

    EFSA Panel on Animal Health and Welfare; More, Simon J.; Bøtner, Anette

    2017-01-01

    prevention and control rules as in Annex IV and Article 8 on the list of animal species related to paratuberculosis. The assessment has been performed following a methodology composed of information collection and compilation, expert judgement on each criterion at individual and, if no consensus was reached...... performed, paratuberculosis can be considered eligible to be listed for Union intervention as laid down in Article 5(3) of the AHL. The disease would comply with the criteria in Sections 3, 4 and 5 of Annex IV of the AHL, for the application of the disease prevention and control rules referred to in points......Paratuberculosis has been assessed according to the criteria of the Animal Health Law (AHL), in particular criteria of Article 7 on disease profile and impacts, Article 5 on the eligibility of paratuberculosis to be listed, Article 9 for the categorisation of paratuberculosis according to disease...

  15. Molecular characterization and drug susceptibility profile of a Mycobacterium avium subspecies avium isolate from a dog with disseminated infection.

    Science.gov (United States)

    Armas, Federica; Furlanello, Tommaso; Camperio, Cristina; Trotta, Michele; Novari, Gianluca; Marianelli, Cinzia

    2016-01-12

    Mycobacterium avium complex (MAC) infections have been described in many mammalian species including humans and pets. We isolated and molecularly typed the causative agent of a rare case of disseminated mycobacteriosis in a dog. We identified the pathogen as a M. avium subspecies avium by sequencing the partial genes gyrB and rpsA. Considering the zoonotic potential of this infection, and in an attempt to ensure the most effective treatment for the animal, we also determined the drug susceptibility profile of the isolate to the most common drugs used to treat MAC disease in humans. The pathogen was tested in vitro against the macrolide clarithromycin, as well as against amikacin, ciprofloxacin, rifampicin, ethambutol and linezolid by the resazurin microdilution assay. It was found to be sensitive to all tested drugs save ethambutol. Despite the fact that the pathogen was sensitive to the therapies administered, the dog's overall clinical status worsened, and the animal died shortly after antimicrobial susceptibility results became available. Nucleotide sequencing of the embB gene, the target gene most commonly associated with ethambutol resistance, showed new missense mutations when compared to sequences available in public databases. In conclusion, we molecularly identified the MAC pathogen and determined its drug susceptibility profile in a relatively short period of time (seven days). We also characterized new genetic mutations likely to have been involved in the observed ethambutol resistance. Our results confirm the usefulness of both the gyrB and the rpsA genes as biomarkers for an accurate identification and differentiation of MAC pathogens.

  16. 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.

  17. PENGEMBANGAN SSP TEMATIK INTEGRATIF UNTUK MEMBANGUN KARAKTER KEJUJURAN DAN KEPEDULIAN SISWA SD KELAS II

    Directory of Open Access Journals (Sweden)

    Sri Hariyati Qodriyah

    2015-07-01

    Full Text Available Penelitian ini bertujuan untuk menghasilkan perangkat pembelajaran berupa SSP Tematik yang dapat mengembangkan karakter siswa kelas 2 sekolah dasar, meliputi karakter kejujuran dan kepedulian. Penelitian ini merupakan penelitian dan pengembangan yang terdiri dari tujuh tahap, yaitu studi pendahuluan, perencanaan, mengembangkan produk awal, uji coba awal, revisi produk utama, uji coba lapangan, dan revisi terhadap produk operasional. SSP yang dikembangkan dievaluasi oleh seorang ahli materi dan media untuk mengetahui validitasnya. Subjek uji coba berjumlah 90 siswa terdiri dari 23 siswa SD Sonosewu, Kasihan, Bantul, DIY sebagai subjek uji coba terbatas dan 33 siswa untuk kelas control dan 34 siswa untuk kelas eksperimen SD 1 Kadipiro, Kasihan, Bantul, DIY. Hasil penelitian ini berupa SSP yang meliputi: silabus, RPP, LKS, dan lembar penilaian. Hasil evaluasi dari ahli materi dan ahli media untuk menguji tingkat kevalidan SSP menyatakan bahwa SSP yang dikembangkan adalah valid dan berkategori “baik”. Hasil uji coba menunjukkan bahwa SSP yang dikembangkan dinyatakan layak serta memenuhi kriteria praktis dan efektif dalam mengembangkan karakter siswa. Pembelajaran dengan SSP yang dikembangkan mampu mengembangkan karakter kejujuran dan kepedulian. Kata Kunci: SSP, karakter jujur dan peduli   DEVELOPING THEMATIC INTEGRATIVE SSP FOR BUILDING THE CHARACTERS OF HONESTY AND CARE GRADE II STUDENT OF ELEMENTARY SCHOOL Abstract This study aims to produce integrated learning sets in the form of thematic SSP that can develop the characters of honesty and care of grade II students of elementary school. This was a research and development study consisting of seven steps, i.e. preliminary study, planning, preliminary product development, preliminary tryout, main product revision, field tryout, revision of operational product, operational product revision, and revision of the final product. The developed SSP was evaluated by a science subject expert to assess

  18. Essential oil composition and antibacterial activity of Tanacetum argenteum (Lam.) Willd. ssp. argenteum and T. densum (Lab.) Schultz Bip. ssp. amani heywood from Turkey.

    Science.gov (United States)

    Polatoğlu, Kaan; Demirci, Fatih; Demirci, Betül; Gören, Nezhun; Başer, Kemal Hüsnü Can

    2010-01-01

    Water-distilled essential oils from aerial parts of Tanacetum argenteum ssp. argenteum and T. densum ssp. amani from Turkey were analyzed by GC and GC/MS. The essential oil of T. argenteum ssp. argenteum was characterized with alpha-pinene 36.7%, beta-pinene 27.5% and 1,8-cineole 9.8%. T. densum ssp. amani was characterized with beta-pinene 27.2%, 1,8-cineole 13.1%, alpha-pinene 9.7% and p-cymene 8.9%. Antibacterial activity of the oils were evaluated for five Gram-positive and five Gram-negative bacteria by using a broth microdilution assay. The highest inhibitory activity was observed against Bacillus cereus for T. argenteum ssp. argenteum oil (125 microg/mL) when compared with positive control chloramphenicol it showed the same inhibition potency. However, the same oil showed lower inhibitory activity against B. subtilis when compared. The oil of T. densum ssp. amani did not show significant activity against the tested microorganisms. DPPH radical scavenging activity of the T. argenteum ssp. argenteum oil was investigated for 15 and 10 mg/mL concentrations. However, the oil did not show significant activity when compared to positive control alpha-tocopherol. Both oils showed toxicity to Vibrio fischeri in the TLC-bioluminescence assay.

  19. Clinical outbreak of Bordetella avium infection in two turkey breeder flocks.

    Science.gov (United States)

    Kelly, B J; Ghazikhanian, G Y; Mayeda, B

    1986-01-01

    An acute upper respiratory disease was observed in two broad-breasted white (BBW) turkey primary breeder flocks. Associated clinical signs included sneezing, depression, and a deep dry cough originating from large conducting airways. Morbidity reached approximately 15-20% of the hens in an affected house. None of the turkeys died, and total feed consumption was not affected. A minimal effect upon egg production was noticed. Sera from an acutely affected flock exhibited a marked rise in titer to Bordetella avium compared with preinfection sera samples. In Case 1, B. avium was isolated in pure culture from affected birds. In Case 2, B. avium was diagnosed by serological results and clinical signs; bacteriological examination was not attempted. The findings presented here are consistent with an acute clinical outbreak of B. avium-induced turkey rhinotracheitis (turkey coryza) in BBW turkey breeder hens.

  20. 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...

  1. Lymphadenitis in children is caused by Mycobacterium avium hominissuis and not related to 'bird tuberculosis'

    NARCIS (Netherlands)

    Bruijnesteijn van Coppenraet, L.E.S.; de Haas, P.E.W.; Lindeboom, J.A.; Kuijper, E.J.; van Soolingen, D.

    2008-01-01

    Mycobacterium avium is the most commonly encountered mycobacterium species among non-Mycobacterium tuberculosis complex (nontuberculous mycobacteria) isolates worldwide and frequently causes lymphadenitis in children. During a multi-centre study in The Netherlands that was performed to determine the

  2. Analysis of CD4+ Th1 cells in immunity to intracellular Mycobacterium avium infection

    OpenAIRE

    Bakke, Marit Øien

    2013-01-01

    Mycobacterial infections are considered a major health problem internationally. Mycobacterium tuberculosis (Mtb), the infectious agent causing tuberculosis, kills 1.4 million people each year. Other non-tuberculous species such as Mycobacterium avium (M. avium) are less pathogenic but cause opportunistic infections mainly in immune-compromised people. The current treatment for different mycobacterial infections is elaborate, expensive and ineffective and new targets for treatment are needed. ...

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

    Directory of Open Access Journals (Sweden)

    Evelyn Guirado

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

  4. Serological comparison of selected isolates of Aeromonas salmonicida ssp. Salmonicida

    Science.gov (United States)

    Hahnel, G.B.; Gould, R.W.; Boatman, E.S.

    1983-01-01

    Eight isolates of Acronionus salmonicida ssp. salmonicida were collected during furunculosis epizootics in North American Pacific coast states and provinces. Both virulent and avirulent forms of each isolate, confirmed by challenge and electron microscopy, were examined. Serological comparisons by cross-absorption agglutination tests revealed no serological differences between isolates. Using the double diffusion precipitin test, a single band was observed when antigen from a sonicated virulent strain was reacted with antiserum against a sonicated, virulent strain absorbed with homologous, avirulent strain. The presence of the single band was eliminated by excess sonication.

  5. VizieR Online Data Catalog: SSP in NIR. II. Synthesis models (Meneses-Goytia+, 2015)

    NARCIS (Netherlands)

    Meneses-Goytia, S.; Peletier, R. F.; Trager, S. C.; Vazdekis, A.

    2015-01-01

    The present Single Stellar Populations (SSP) models are derived from my Ph.D.'s thesis and this paper. The following nomenclature is used throughout the paper and the website (http://smg.astro-research.net/ssp-models/the-models/) to describe the models, e.g. MarS models use the M08 isochrones (Mar)

  6. Kvalitet, faglighed og legitime videnformer i SSP-samarbejdet under transformation

    DEFF Research Database (Denmark)

    Tamborg, Andreas Lindenskov

    2016-01-01

    Quality, professionalism and legitimate knowledge in a changing SSP-collaboration. This article explores the inter professional collaboration in a crime preventive initiative called SSP, which is a collaboration between school, social work and police. By drawing on interviews of actors from...

  7. 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.

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

    Science.gov (United States)

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

    2011-11-01

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

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

    Directory of Open Access Journals (Sweden)

    Kim Woo

    2006-10-01

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

  10. Enhanced radiometric detection of Mycobacterium paratuberculosis by using filter-concentrated bovine fecal specimens

    International Nuclear Information System (INIS)

    Collins, M.T.; Kenefick, K.B.; Sockett, D.C.; Lambrecht, R.S.; McDonald, J.; Jorgensen, J.B.

    1990-01-01

    A commercial radiometric medium, BACTEC 12B, was modified by addition of mycobactin, egg yolk suspension, and antibiotics (vancomycin, amphotericin B, and nalidixic acid). Decontaminated bovine fecal specimens were filter concentrated by using 3-microns-pore-size, 13-mm-diameter polycarbonate filters, and the entire filter was placed into the radiometric broth. Comparison of the radiometric technique with conventional methods on 603 cattle from 9 Mycobacterium paratuberculosis-infected herds found that of 75 positive specimens, the radiometric technique detected 92% while conventional methods detected 60% (P less than 0.0005). Only 3.9% of radiometric cultures were contaminated. To measure the effect of filter concentration of specimens on the detection rate, 5 cattle with minimal and 5 with moderate ileum histopathology were sampled weekly for 3 weeks. M. paratuberculosis was detected in 33.3% of nonfiltered specimens and 76.7% of filtered specimens (P less than 0.005). Detection rates were directly correlated with the severity of disease, and the advantage of specimen concentration was greatest on fecal specimens from cattle with low-grade infections. Detection times were also correlated with infection severity: 13.4 +/- 5.9 days with smear-positive specimens, 27.9 +/- 8.7 days with feces from cows with typical subclinical infections, and 38.7 +/- 3.8 days with fecal specimens from cows with low-grade infections. Use of a cocktail of vancomycin, amphotericin B, and nalidixic acid for selective suppression of nonmycobacterial contaminants was better than the commercial product PANTA (Becton Dickinson Microbiologic Systems, Towson, Md.) only when specimens contained very low numbers of M. paratuberculosis

  11. 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. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. 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.

  13. Essential-oil composition of Daucus carota ssp. major (Pastinocello Carrot) and nine different commercial varieties of Daucus carota ssp. sativus fruits.

    Science.gov (United States)

    Flamini, Guido; Cosimi, Elena; Cioni, Pier Luigi; Molfetta, Ilaria; Braca, Alessandra

    2014-07-01

    The chemical composition of the essential oils obtained by hydrodistillation from the pastinocello carrot, Daucus carota ssp. major (Vis.) Arcang. (flowers and achenes), and from nine different commercial varieties of D. carota L. ssp. sativus (achenes) was investigated by GC/MS analyses. Selective breeding over centuries of a naturally occurring subspecies of the wild carrot, D. carota L. ssp. sativus, has produced the common garden vegetable with reduced bitterness, increased sweetness, and minimized woody core. On the other hand, the cultivation of the pastinocello carrot has been abandoned, even if, recently, there has been renewed interest in the development of this species, which risks genetic erosion. The cultivated carrot (D. carota ssp. sativus) and the pastinocello carrot (D. carota ssp. major) were classified as different subspecies of the same species. This close relationship between the two subspecies urged us to compare the chemical composition of their essential oils, to evaluate the differences. The main essential-oil constituents isolated from the pastinocello fruits were geranyl acetate (34.2%), α-pinene (12.9%), geraniol (6.9%), myrcene (4.7%), epi-α-bisabolol (4.5%), sabinene (3.3%), and limonene (3.0%). The fruit essential oils of the nine commercial varieties of D. carota ssp. sativus were very different from that of pastinocello, as also confirmed by multivariate statistical analyses. Copyright © 2014 Verlag Helvetica Chimica Acta AG, Zürich.

  14. 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.

  15. The persistence of Mycobacterium avium in a drinking water system, what is the risk to human health?

    Science.gov (United States)

    Drinking water is believed to be a major source of human exposure to nontuberculous mycobacteria (NTM) such as Mycobacterium avium. We monitored the prevalence of M. avium in a drinking water system during the addition of filtration treatment. Our goal was to determine if the pre...

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

    Science.gov (United States)

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

  17. [Usefulness of the variable numbers of tandem repeats (VNTR) analysis for complex infections of Mycobacterium avium and Mycobacterium intracellulare].

    Science.gov (United States)

    Tsunematsu, Noriko; Goto, Mieko; Saiki, Yumiko; Baba, Michiko; Udagawa, Tadashi; Kazumi, Yuko

    2008-09-01

    The bacilli which were isolated from a patient suspected of the mixed infections with Mycobacterium avium and Mycobacterium intracellulare, were analyzed. The genotypes of M. avium in the sedimented fractions of treated sputum and in some colonies isolated from Ogawa medium were compared by the Variable Numbers of Tandem Repeats (VNTR). A woman, aged 57. Mycobacterial species isolated from some colonies by culture in 2004 and 2006 and from the treated sputum in 2006, were determined by DNA sequencing analysis of the 16S rRNA gene. Also, by using VNTR, the genotype of mycobacteria was analyzed. [Results] (1) The colony isolated from Ogawa medium in 2004 was monoclonal M. avium. (2) By VNTR analyses of specimens in 2006, multiple acid-fast bacteria were found in the sputum sediment and in isolated bacteria from Ogawa medium. (3) By analyses of 16S rRNA DNA sequence, M. avium and M. intracellulare were found in the colonies isolated from the sputum sediment and the Ogawa medium in 2006. (4) The same VNTR patterns were obtained in M. avium in 2004 and 2006 when single colony was analyzed. (5) From the showerhead and culvert of the bathroom in the patient's house, M. avium was not detected. By VNTR analyses, it was considered that the mixed infections of M. avium and M. intracellulare had been generated during treatment in this case. Therefore, in the case of suspected complex infection, VNTR analysis would be a useful genotyping method in M. avium complex infection.

  18. Merging DNA typing and network analysis to assess the transmission of paratuberculosis between farms.

    Science.gov (United States)

    Marquetoux, N; Heuer, C; Wilson, P; Ridler, A; Stevenson, M

    2016-11-01

    Paratuberculosis, a chronic enteric infection caused by Mycobacterium subsp. paratuberculosis (MAP), is endemic in all farmed ruminant species in New Zealand. The use of genotyping in combination with network analysis of livestock movement events from one farm location to another has the potential to contribute to our understanding of between-farm transmission events. We studied a population of 122 farms from a corporate commercial livestock enterprise in New Zealand, trading with each other in near isolation from other commercial farms. The data consisted of longitudinal movements to and from these farms between 2006 and 2010, as well as the results of cross-sectional MAP screening and genotyping performed in 2010. We explored associations between past livestock movements and current strain type distribution in this population of farms using quadratic assignment procedure. Our results show that measures of farm clustering within the movement network were significantly associated with sharing of MAP strains. For example, farms closely related by trade were twice as likely to share the same strains of MAP (p=0.033). Other covariates were also associated with the probability of sharing the same strains of MAP, such as being located on the same island (OR=5.8 to 8.7, pfarming the same livestock species and Euclidian distance between farms. The novel approach we used supports the hypothesis that livestock movement is indeed a significant contributor to farm-to-farm transmission of MAP. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Surveillance of bulk raw and commercially pasteurized cows' milk from approved Irish liquid-milk pasteurization plants to determine the incidence of Mycobacterium paratuberculosis.

    Science.gov (United States)

    O'Reilly, Ciara E; O'Connor, Lisa; Anderson, Wayne; Harvey, Peter; Grant, Irene R; Donaghy, John; Rowe, Michael; O'Mahony, Pat

    2004-09-01

    Over the 13-month period from October 2000 to November 2001 (inclusive), the Food Safety Authority of Ireland (FSAI) carried out surveillance of Irish bulk raw (n = 389) and commercially pasteurized (n = 357) liquid-milk supplies to determine the incidence of Mycobacterium paratuberculosis. The pasteurization time-temperature conditions were recorded for all pasteurized samples. Overall, 56% of whole-milk pasteurized samples had been heat treated at or above a time-temperature combination of 75 degrees C for 25 s. All analyses were undertaken at the Department of Food Science (Food Microbiology) laboratory at Queen's University Belfast. Each milk sample was subjected to two tests for M. paratuberculosis: immunomagnetic separation-PCR (IMS-PCR; to detect the presence of M. paratuberculosis cells, live or dead) and chemical decontamination and culture (to confirm the presence of viable M. paratuberculosis). Overall, M. paratuberculosis DNA was detected by IMS-PCR in 50 (12.9%; 95% confidence interval, 9.9 to 16.5%) raw-milk samples and 35 (9.8%; 95% confidence interval, 7.1 to 13.3%) pasteurized-milk samples. Confirmed M. paratuberculosis was cultured from one raw-milk sample and no pasteurized-milk samples. It is concluded that M. paratuberculosis DNA is occasionally present at low levels in both raw and commercially pasteurized cows' milk. However, since no viable M. paratuberculosis was isolated from commercially pasteurized cows' milk on retail sale in the Republic of Ireland, current pasteurization procedures are considered to be effective.

  20. Natural killer cell activities of synbiotic Lactobacillus casei ssp. casei in conjunction with dextran.

    Science.gov (United States)

    Ogawa, T; Asai, Y; Tamai, R; Makimura, Y; Sakamoto, H; Hashikawa, S; Yasuda, K

    2006-01-01

    We have reported previously that Lactobacillus casei ssp. casei, together with specific substrate dextran, exhibited an adjuvant effect of stimulating humoral immune responses against bovine serum albumin (BSA) as a model antigen in BALB/c mice. In the present study, among the Lactobacillus species tested, L. casei ssp. casei with dextran significantly elevated the natural killer (NK) cell activities in spleen mononuclear cells from BALB/c mice in comparison to L. casei ssp. casei alone or other Lactobacillus species with or without dextran. Oral administration of L. casei ssp. casei together with dextran also resulted in a significant increase of NK cell activities in healthy human volunteers. Further, L. casei ssp. casei induced significant production of interleukin (IL)-12 in human peripheral blood mononuclear cells and IL-15 mRNA expression in the human intestinal epithelial cell line Caco-2. L. casei ssp. casei with dextran in food also significantly elevated the survival rate of BALB/c mice bearing Meth-A cells. Taken together, these results demonstrate that dietary synbiotic supplementation which is a combination of the L. casei ssp. casei used as a probiotic together with the dextran, a specific substrate as a prebiotic, efficiently elicits murine and human NK cell activities.

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

    Directory of Open Access Journals (Sweden)

    Gupta K

    2010-01-01

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

  2. A validity study of the SSP-School Inclusion Questionnaire

    Directory of Open Access Journals (Sweden)

    DIANA DĂMEAN

    2012-12-01

    Full Text Available The paper is based on the results of a research that aimed to identify the factors influencing school inclusion among disadvantagedstudents. The main outcome of the research was the development of the SSP-School Inclusion Questionnaire, intended to: (1measure school inclusion levels among students; (2 identify students with significantly lower levels of school inclusion, whoneed to be included in intervention programs; and (3 measure the impact of intervention programs using subjective indicators(beneficiaries’ perception of school and teachers, feelings of safety in school, school performance. The questionnaire was completedby 480 vocational school students aged 15-19 from a large developed urban area (Cluj-Napoca in Romania. The purpose of theresearch was to test the validity and reliability of the instrument and to propose it to professionals working in education (schoolpsychologists, school counsellors etc. for efficient data collection and for measuring the impact of educational intervention.

  3. Functional characterization of gynodioecy in Fragaria vesca ssp. bracteata (Rosaceae).

    Science.gov (United States)

    Li, Junmin; Koski, Matthew H; Ashman, Tia-Lynn

    2012-02-01

    Gynodioecy is a phylogenetically widespread and important sexual system where females coexist with hermaphrodites. Because dioecy can arise from gynodioecy, characterization of gynodioecy in close relatives of dioecious and sub-dioecious species can provide insight into this transition. Thus, we sought to determine whether Fragaria vesca ssp. bracteata, a close relative to F. chiloensis and F. virginiana, exhibits the functional and population genetic hallmarks of a gynodioecious species. We compared reproductive allocation of females and hermaphrodites grown in the greenhouse and estimated genetic diversity (allelic diversity, heterozygosity) and inbreeding coefficients for field-collected adults of both sexes using simple sequence repeat (SSR) markers. We estimated mating system and early seed fitness from open-pollinated families of both sex morphs. Under greenhouse conditions, females and hermaphrodites allocated similarly to all reproductive traits except flower number, and, as a consequence, females produced 30 % fewer seeds per plant than hermaphrodites. Under natural conditions, hermaphrodites produce seeds by self-fertilization approx. 75 % of the time, and females produced outcrossed seeds with very little biparental inbreeding. Consistent with inbreeding depression, seeds from open-pollinated hermaphrodites were less likely to germinate than those from females, and family-level estimates of hermaphrodite selfing rates were negatively correlated with germination success and speed. Furthermore, estimates of inbreeding depression based on genetic markers and population genetic theory indicate that inbreeding depression in the field could be high. The joint consideration of allocation and mating system suggests that compensation may be sufficient to maintain females given the current understanding of sex determination. Fragaria vesca ssp. bracteata exhibited similar sex morph-dependent patterns of mating system and genetic diversity, but less reproductive

  4. 12038_2016_9599_Article_Supplementary 1..12

    Indian Academy of Sciences (India)

    Actinobacteria Mammal. Tuberculosis type pulmonary infections. 20. Mycobacterium avium subsp. paratuberculosis K-10. Actinobacteria Mammal. Paratuberculosis. 21. Mycobacterium bovis AF2122/97. Actinobacteria Human, Animal Tuberculosis in cattle. 22. Mycobacterium bovis BCG str. Mexico. Actinobacteria Bovine.

  5. 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

  6. Disseminated mycobacteriosis due to Mycobacterium avium in captive Bengal tiger (Panthera tigris).

    Science.gov (United States)

    Cho, Ho-Seong; Kim, Yong-Hwan; Park, Nam-Yong

    2006-05-01

    A 2-year-old captive female Bengal Tiger (Panthera tigris) died after prolonged anorexia in the Gwangju Uchi Park Zoo, Gwangju, Republic of Korea. Necropsy revealed multiple nodules of varying sizes in the lung, liver, kidney, and spleen. Histopathologic examination revealed a typical granuloma composed of caseous necrotic areas surrounded by lymphocytes with a few giant cells and foamy macrophages. Periodic acid-Schiff stain and Gomori methenamine silver stain did not reveal any fungal bodies. The Ziehl-Neelsen acid-fast stain revealed few acid-fast organisms in the lung, liver, kidney, and spleen. A polymerase chain reaction assay of the lung, liver, kidney, and spleen yielded a positive result for Mycobacterium avium subsp. avium. This is an unusual case of disseminated infection of a wild mammal with avian mycobacteriosis, and is believed to be most likely associated with the feeding of tigers with culled chickens infected with M. avium.

  7. 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.

  8. Hypersensitivity pneumonitis with Mycobacterium avium complex among spa workers.

    Science.gov (United States)

    Moraga-McHaley, Stephanie Ann; Landen, Michael; Krapfl, Heidi; Sewell, C Mack

    2013-01-01

    The New Mexico Department of Health (NMDOH) investigated the cause of two cases of hypersensitivity pneumonitis (HP) in spa maintenance workers with laboratory confirmed Mycobacterium avium complex (MAC). The investigation occurred in tandem with worker protection and swimming pool regulatory investigations by the New Mexico Environment Department at the spa where the workers were employed. The investigation was conducted in order to identify unreported cases, exposure source(s), and to prevent further worker exposure. NMDOH surveyed 57 spa employees about symptoms and exposures, categorized jobs according to self-reported exposure to water, and computed odds ratios for symptom reporting by exposure category. Environmental isolates from spa water and filter swabs were cultured and compared to patient isolates by the Environmental and Applied Microbiology Team, Centers for Disease Control and Prevention (CDC). Workers with the highest exposure reported more HP-like symptoms (OR = 9.6), as did intermediate exposure workers (OR = 6.5), compared to workers with no aerosolized water exposure. Two of 13 environmental isolates were closely related to one of the patient isolates. Workers were likely exposed during spray cleaning of cartridge filters in a poorly ventilated work space. Recommendations include inhibiting organism growth in spa systems, assuring the use of respiratory protection, and adequately ventilating work spaces where filters and equipment are cleaned.

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

    DEFF Research Database (Denmark)

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

    2017-01-01

    Introduction: The aim of this study was to test a commercial bovine enzyme-linked immunosorbent assay for investigating antibody activity against Mycobacterium avium complex. Methods: All patients at the Copenhagen Cystic Fibrosis (CF) Center who had culture for nontuberculous mycobacteria......, corresponding to a sensitivity of 100% (54–100), specificity of 66% (60–72), a positive predictive value of 6% (2–13), and negative predictive value of 100% (98–100). Conclusion: While not suited for direct diagnosis of M. avium complex due to a high number of false positive subjects, the assay proved useful...

  10. Biodegradation of naphthalene by Bordetella avium isolated from petroleum refinery wastewater in Egypt and its pathway

    Directory of Open Access Journals (Sweden)

    M.A.M. Abo-State

    2018-01-01

    The results revealed that isolate MAM-P22 was the best Naphthalene degrader. It degraded 95% of the highest concentration 7 mM. The best Naphthalene degrader bacterial Isolate MAM-22 was identified by 16S-rRNA showed a similarity of 98% to Bordetella avium strain with accession No. 041769.1. The results of GC/MS analysis revealed that Bordetella avium MAM-P22 degraded Naphthalene to give six intermediate compounds, These compounds were 1,2-Benzene dicarboxylic acid, Butyl-2,4-dimethyl-2-nitro-4-pentenoate, 1-Nonen-3-ol, Eicosane, Nonacosane.

  11. An abstract class loader for the SSP and its implementation in TL.

    Energy Technology Data Exchange (ETDEWEB)

    Wickstrom, Gregory Lloyd; Winter, Victor Lono (University of Nebraska at Omaha, Omaha, NE); Fraij, Fares (University of Texas at El Paso, El Paso, TX); Roach, Steve (University of Texas at El Paso, El Paso, TX); Beranek, Jason (University of Nebraska at Omaha, Omaha, NE)

    2004-08-01

    The SSP is a hardware implementation of a subset of the JVM for use in high consequence embedded applications. In this context, a majority of the activities belonging to class loading, as it is defined in the specification of the JVM, can be performed statically. Static class loading has the net result of dramatically simplifying the design of the SSP as well as increasing its performance. Due to the high consequence nature of its applications, strong evidence must be provided that all aspects of the SSP have been implemented correctly. This includes the class loader. This article explores the possibility of formally verifying a class loader for the SSP implemented in the strategic programming language TL. Specifically, an implementation of the core activities of an abstract class loader is presented and its verification in ACL2 is considered.

  12. ICLUS v2.1 land use projections for the Fourth National Climate Assessment (SSP2)

    Data.gov (United States)

    U.S. Environmental Protection Agency — SSP2 is a “middle-of-the-road” projection, where social, economic and technological trends do not shift markedly from historical patterns, resulting in a U.S....

  13. Detection of Bordetella avium by TaqMan real-time PCR in tracheal swabs from wildlife birds.

    Science.gov (United States)

    Stenzel, T; Pestka, D; Tykałowski, B; Śmiałek, M; Koncicki, A; Bancerz-Kisiel, A

    2017-03-28

    Bordetella avium, the causing agent of bordetellosis, a highly contagious infection of the respiratory tract in young poultry, causes significant losses in poultry farming throughout the world. Wildlife birds can be a reservoir of various pathogens that infect farm animals. For this reason the studies were conducted to estimate the prevalence of Bordetella avium in wildlife birds in Poland. Tracheal swab samples were collected from 650 birds representing 27 species. The bacterial DNA was isolated directly from the swabs and screened for Bordetella avium by TaqMan real-time PCR. The assay specificity was evaluated by testing DNA isolated from 8 other bacteria that can be present in avian respiratory tract, and there was no amplification from non-Bordetella avium agents. Test sensitivity was determined by preparing standard tenfold serial dilutions of DNA isolated from positive control. The assay revealed to be sensitive, with detection limit of approximately 4.07x10^2 copies of Bordetella avium DNA. The genetic material of Bordetella avium was found in 54.54% of common pheasants, in 9.09% of Eurasian coots, in 3.22% of black-headed gulls and in 2.77% of mallard ducks. The results of this study point to low prevalence of Bordetella avium infections in wildlife birds. The results also show that described molecular assay proved to be suitable for the rapid diagnosis of bordetellosis in the routine diagnostic laboratory.

  14. ORF Alignment: NC_002944 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available gth = 489 ... Query: 20 ... WWTPDTLGDLLARGLKDNQHNTFRVHSAVRPFAGTFGDVEXXXXXXXXXXXXXXVGPGDV 79 ... WWTP...DTLGDLLARGLKDNQHNTFRVHSAVRPFAGTFGDVE ... VGPGDV Sbjct: 1 ... WWTPDTLGDLLARGLKDNQ... ... protein MAP2188c [Mycobacterium avium subsp. ... paratuberculosis str. k10] ... Len

  15. Development of a Multi-Stage Vaccine against Paratuberculosis in Cattle

    DEFF Research Database (Denmark)

    Thakur, Aneesh

    of quantitative real-time PCR (qPCR) for the evaluation of microbial load in tissues and vaccine efficacy (Article 4). Costimulation of vaccine-induced ex vivo T cells significantly increased IFN-γ levels following use of anti-CD28 and anti-CD49d antibodies (Article 2). Recombinant interleukin IL-12 (rIL-12) also...... to 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...

  16. Multi-stage subunit vaccine development against Mycobacterium paratuberculosis and Johne’s disease in ruminants

    DEFF Research Database (Denmark)

    Jungersen, Gregers

    paratuberculosis provide only partial protection and interfere with diagnostic tests for JD and surveillance for bovine TB. In contrast, recombinant subunit vaccines can be designed to be used without compromising control of bTB and Map. Taking advantage of data from mouse TB studies, and early Map vaccination......- and field-studies we developed a vaccine with a single recombinant fusion protein comprising four acute-stage antigens (Ags) and one latent-stage Ag formulated in adjuvant (FET-vaccine). In post-exposure vaccination of calves and goats with necropsy 8-12 months post inoculation, we determined...... in macrophages. The disease progression is very slow with neonatal animals being the most susceptible to infection, but without development of detectable IFN-γ responses for months after infection and rarely with clinical disease before the second or third year of life. Available whole cell vaccines against...

  17. Continuous-data diagnostic tests for paratuberculosis as a multistage disease

    DEFF Research Database (Denmark)

    Toft, Nils; Nielsen, Søren Saxmose; Jørgensen, Erik

    2005-01-01

    We devised a general method for interpretation of multistage diseases using continuous-data diagnostic tests. As an example, we used paratuberculosis as a multistage infection with 2 stages of infection as well as a noninfected state. Using data from a Danish research project, a fecal culture...... testing scheme was linked to an indirect ELISA and adjusted for covariates (parity, age at first calving, and days in milk). We used the log-transformed optical densities in a Bayesian network to obtain the probabilities for each of the 3 infection stages for a given optical density (adjusted...... for covariates). The strength of this approach was that the uncertainty associated with a test was imposed directly on the individual test result rather than aggregated into the population-based measures of test properties (i.e., sensitivity and specificity)...

  18. 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.

  19. Inactivation of Mycobacterium paratuberculosis and Mycobacterium tuberculosis in fresh soft cheese by gamma radiation

    International Nuclear Information System (INIS)

    Badr, Hesham M.

    2011-01-01

    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 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.

  20. Full Scale Test of a SSP 34m boxgirder 2

    DEFF Research Database (Denmark)

    Jensen, Find Mølholt; Branner, Kim; Nielsen, Per Hørlyk

    was part of a proof of concept investigation for a patent. The tests were performed at the Blaest test facility in August 2007. The tests are an important part of a research project established in cooperation between Risø National Laboratory for sustainable energy – Technical university of Denmark, SSP......This report presents the setup and result from three static full-scale tests of the reinforced glass fiber/epoxy box girder used in a 34m wind turbine blade. One test was without reinforcement one with cap reinforcement and the final test was with rib reinforcement. The cap reinforcement test......-Technology A/S and Blaest (Blade test centre A/S) and it has been performed as a part of Find Mølholt Jensen’s PhD thesis. This report is the second data report containing the complete test data for the three full-scale tests. This report deals only with the test methods and the obtained results...

  1. 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.

  2. 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

  3. 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")

  4. Growth and fruit bearing of the sweet cherry (Prunus avium L

    African Journals Online (AJOL)

    Radunic

    2011-06-06

    Jun 6, 2011 ... Modern intensive production of sweet cherry (Prunus avium L.) tends to planting of high quality cultivars on the dwarfing rootstocks in high density orchards. The most productive training system is used, providing an ideal condition for undisturbed growth and yield. The main objective of this study.

  5. COMPARISON OF MYCOBACTERIUM AVIUM ISOLATES FROM DRINKING WATER AND FROM THE POPULATION SERVED BY THE SYSTEM

    Science.gov (United States)

    Background: Current evidence suggests that drinking water, soil, and produce are potential sources of Mycobacterium avium infections, a pathogen not known to be transmitted person-to-person. Methods: We sampled water during 2000 - 2002 from a large municipal drinking wate...

  6. Improved detection of Mycobacterium avium complex with the Bactec radiometric system

    International Nuclear Information System (INIS)

    Hoffner, S.E.

    1988-01-01

    A reconsideration of the laboratory methods used for primary isolation of mycobacteria other than Mycobacterium tuberculosis is needed due to the increasingly recognized importance of such mycobacterial infections in immunocompromised patients. One example of this is the severe opportunistic infections caused by Mycobacterium avium complex among AIDS patients. In this study, the Bactec radiometric system was compared to conventional culture on solid medium for the detection of M. avium complex in 3,612 selected clinical specimens, mainly of extrapulmonary origin. Of a total number of 63 M. avium complex isolates, the Bactec system detected 58 (92%), compared to 37 (59%) for conventional culture. A much more rapid detection was attained with radiometric technique than with conventional culture. The mean detection time for the cultures positive with both methods was 7.1 and 28.3 days, respectively. The Bactec radiometric system achieves a rapid and significantly more sensitive detection and seems to be an excellent complement to conventional culture in the laboratory diagnosis of infections with the M. avium complex

  7. Shared Mycobacterium avium genotypes observed among unlinked clinical and environmental isolates

    Science.gov (United States)

    Our understanding of the sources of Mycobacterium avium infection is partially based on genotypic matching of pathogen isolates from cases and environmental sources. These approaches assume that genotypic identity is rare in isolates from unlinked cases or sources. To test this, ...

  8. Shared Mycobacterium avium genotypes observed among unlinked clinical and environmental isolates*

    Science.gov (United States)

    Our understanding of the sources of Mycobacterium avium infection is partially based on genotypic matching of pathogen isolates from cases and environmental sources. These approaches assume that genotypic identity is rare in isolates from unlinked cases or sources. To test this a...

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

    Science.gov (United States)

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

  10. Positioning Space Solar Power (SSP) as the Next Logical Step after the International Space Station (ISS)

    Science.gov (United States)

    Charania, A.

    2002-01-01

    At the end of the first decade of the 21st century, the International Space Station (ISS) will stand as a testament of the engineering capabilities of the international community. The choices for the next logical step for this community remain vast and conflicting: a Mars mission, moon colonization, Space Solar Power (SSP), etc. This examination focuses on positioning SSP as one such candidate for consideration. A marketing roadmap is presented that reveals the potential benefits of SSP to both the space community and the global populace at large. Recognizing that scientific efficiency itself has no constituency large enough to persuade entities to outlay funds for such projects, a holistic approach is taken to positioning SSP. This includes the scientific, engineering, exploratory, economic, political, and development capabilities of the system. SSP can be seen as both space exploration related and a resource project for undeveloped nations. Coupling these two non-traditional areas yields a broader constituency for the project that each one alone could generate. Space exploration is many times seen as irrelevant to the condition of the populace of the planet from which the money comes for such projects. When in this new century, billions of people on the planet still have never made a phone call or even have access to clean water, the origins of this skepticism can be understandable. An area of concern is the problem of not living up to the claims of overeager program marketers. Just as the ISS may never live up to the claims of its advocates in terms of space research, any SSP program must be careful in not promising utopian global solutions to any future energy starved world. Technically, SSP is a very difficult problem, even harder than creating the ISS, yet the promise it can hold for both space exploration and Earth development can lead to a renaissance of the relevance of space to the lives of the citizens of the world.

  11. SSP: an interval integer linear programming for de novo transcriptome assembly and isoform discovery of RNA-seq reads.

    Science.gov (United States)

    Safikhani, Zhaleh; Sadeghi, Mehdi; Pezeshk, Hamid; Eslahchi, Changiz

    2013-01-01

    Recent advances in the sequencing technologies have provided a handful of RNA-seq datasets for transcriptome analysis. However, reconstruction of full-length isoforms and estimation of the expression level of transcripts with a low cost are challenging tasks. We propose a novel de novo method named SSP that incorporates interval integer linear programming to resolve alternatively spliced isoforms and reconstruct the whole transcriptome from short reads. Experimental results show that SSP is fast and precise in determining different alternatively spliced isoforms along with the estimation of reconstructed transcript abundances. The SSP software package is available at http://www.bioinf.cs.ipm.ir/software/ssp. © 2013.

  12. [Distribution of PRA patterns of clinical isolates of the Mycobacterium avium complex from Spain and South America].

    Science.gov (United States)

    Murcia, Martha Isabel; Leao, Sylvia Cardoso; Ritacco, Viviana; Palenque, Elia; de Oliveira, Rosangela Siqueira; Reniero, Ana; Menendez, Maria Carmen; Telles, María Alice da Silva; Hadad, David Jamil; Barrera, Lucía; García, María Jesús

    2004-06-01

    Mycobacterium avium complex (MAC) infections are the most frequent systemic infections associated with advanced AIDS. DNA probes for accurate identification of mycobacteria are available but are very expensive in many Latin American settings. Consequently, most Latin American diagnostic laboratories employ inaccurate and outdated tests for mycobacteria identification. Therefore, PCR restriction analysis (PRA) of the hsp65 gene was evaluated for the identification of 163 MAC human isolates originated from Spain and South America. The predominant PRA type in each country was: M. avium type I in Argentina (23/42, 55%) and Brazil (48/72, 67%), M. avium type II in Spain (18/26, 69%) and M. avium type III in Colombia (10/23, 43%). The Colombia frequency is noteworthy, since the PRA type III was quite infrequent in the other three countries. Furthermore, its presence has not been reported outside the Americas. The advantages and disadvantages of PRA in diagnostic mycobacteriology are discussed.

  13. Evolutionary genomics of grape (Vitis vinifera ssp. vinifera) domestication.

    Science.gov (United States)

    Zhou, Yongfeng; Massonnet, Mélanie; Sanjak, Jaleal S; Cantu, Dario; Gaut, Brandon S

    2017-10-31

    We gathered genomic data from grapes ( Vitis vinifera ssp. vinifera ), a clonally propagated perennial crop, to address three ongoing mysteries about plant domestication. The first is the duration of domestication; archaeological evidence suggests that domestication occurs over millennia, but genetic evidence indicates that it can occur rapidly. We estimated that our wild and cultivated grape samples diverged ∼22,000 years ago and that the cultivated lineage experienced a steady decline in population size ( N e ) thereafter. The long decline may reflect low-intensity management by humans before domestication. The second mystery is the identification of genes that contribute to domestication phenotypes. In cultivated grapes, we identified candidate-selected genes that function in sugar metabolism, flower development, and stress responses. In contrast, candidate-selected genes in the wild sample were limited to abiotic and biotic stress responses. A genomic region of high divergence corresponded to the sex determination region and included a candidate male sterility factor and additional genes with sex-specific expression. The third mystery concerns the cost of domestication. Annual crops accumulate putatively deleterious variants, in part due to strong domestication bottlenecks. The domestication of perennial crops differs from that of annuals in several ways, including the intensity of bottlenecks, and it is not yet clear if they accumulate deleterious variants. We found that grape accessions contained 5.2% more deleterious variants than wild individuals, and these were more often in a heterozygous state. Using forward simulations, we confirm that clonal propagation leads to the accumulation of recessive deleterious mutations but without decreasing fitness. Copyright © 2017 the Author(s). Published by PNAS.

  14. Definitive differentiation between single and mixed mycobacterial infections in red deer (Cervus elaphus) by a combination of duplex amplification of p34 and f57 sequences and Hpy188I enzymatic restriction of duplex amplicons.

    Science.gov (United States)

    Godfroid, Jacques; Delcorps, Cathy; Irenge, Leonid M; Walravens, Karl; Marché, Sylvie; Gala, Jean-Luc

    2005-09-01

    Severe emaciation and mortalities suggestive of mycobacterial infections were recently reported for both adult and young wild red deer (Cervus elaphus) in the southeastern part of Belgium. In deer, tuberculous lesions are not pathognomonic of Mycobacterium bovis infection due to gross and microscopic similarities with lesions caused by Mycobacterium avium subsp. paratuberculosis or M. avium subsp. avium. The aim of this study was to improve molecular methods for the species-specific identification of M. bovis, M. avium subsp. avium, and M. avium subsp. paratuberculosis in mycobacterial infections of deer. DNA banding patterns were assessed prior to and after Hpy188I restriction of f57-upstream (us)-p34 duplex amplicons. The duplex f57-us-p34 PCR differentiated M. bovis from M. avium subsp. paratuberculosis and M. avium subsp. avium infections, whereas the restriction step differentiated single M. avium subsp. paratuberculosis or M. avium subsp. avium infections from mixed M. avium subsp. paratuberculosis/M. avium subsp. avium infections. The endonuclease Hpy188I cleaves DNA between nucleotides N and G in the unique TCNGA sequence. This restriction site was found at position 168 upstream of the us-p34 initiation codon in all M. avium subsp. avium strains tested, regardless of their origin and the results of IS901 PCR. In contrast, the restriction site was abrogated in all M. avium subsp. paratuberculosis strains tested, independent of their origin, Mycobactin J dependency, and IS900 PCR results. Consequently, a two-step strategy, i.e., duplex us-p34-f57 PCR and Hpy188I restriction, allowed us to exclude M. bovis infection and to identify single (M. avium subsp. paratuberculosis or M. avium subsp. avium) or mixed (M. avium subsp. paratuberculosis/M. avium subsp. avium) infections in wild red deer in Belgium. Accordingly, we propose to integrate, in a functional molecular definition of M. avium subsp. paratuberculosis, the absence of the Hpy188I restriction site from

  15. Immune-enhancing effects of Taishan Pinus massoniana pollen polysaccharides on DNA vaccine expressing Bordetella avium ompA

    Directory of Open Access Journals (Sweden)

    Fujie eZhu

    2016-02-01

    Full Text Available Bordetella avium is the causative agent of bordetellosis, which remains to be the cause of severe losses in the turkey industry. Given the lack of vaccines that can provide good protection, developing a novel vaccine against B. avium infection is crucial. In this study, we constructed a eukaryotic expression plasmid, which expressed the outer membrane protein A (ompA of B. avium, to prepare a B. avium recombinant ompA-DNA vaccine. Three concentrations (low, middle, and high of Taishan Pinus massoniana pollen polysaccharides (TPPPS, a known immunomodulator, were used as adjuvants, and their immune conditioning effects on the developed DNA vaccine were examined. The pure ompA-DNA vaccine, Freund's incomplete adjuvant ompA-DNA vaccine, and the empty plasmid served as the controls. The chickens in each group were separately inoculated with these vaccines three times at 1, 7 and 14 days old. Dynamic changes in antibody production, cytokine secretion, and lymphocyte count were then determined from 7 days to 49 days after the first inoculation. Protective rates of the vaccines were also determined after the third inoculation. Results showed that the pure DNA vaccine obviously induced the production of antibodies, the secretion of cytokines, and the increase in CD4+ and CD8+ T lymphocyte counts in peripheral blood, as well as provided a protective rate of 50% to the B. avium-challenged chickens. The chickens inoculated with the TPPPS adjuvant ompA-DNA vaccine and Freund’s adjuvant ompA-DNA vaccine demonstrated higher levels of immune responses than those inoculated with pure ompA-DNA vaccine, whereas only the ompA-DNA vaccine with 200 mg/mL TPPPS completely protected the chickens against B. avium infection. These findings indicate that the B. avium ompA-DNA vaccine combined with TPPPS is a potentially effective B. avium vaccine.

  16. Clinical significance and epidemiologic analyses of Mycobacterium avium and Mycobacterium intracellulare lung disease from post-marketing surveillance.

    Science.gov (United States)

    Suzuki, Katsuhiro; Kurashima, Atsuyuki; Tatsuno, Kinji; Kadota, Jun-Ichi

    2018-01-01

    In Japan, nontuberculous mycobacterial lung disease is mostly attributable to Mycobacterium avium complex (MAC), i.e., M. avium or M. intracellulare. However, clinical features of the disease caused by these two pathogens have not been studied sufficiently yet. A post-marketing survey of clarithromycin was performed at 130 facilities across Japan. The data on patients with M. avium infection and patients with M. intracellulare infection were selected from this survey for comparison of background variables and clinical features of the two pathogens. Among the patients analyzed (n = 368), 67.4% had M. avium infection and 32.6% had M. intracellulare infection. Stratified analysis revealed no significant differences between the ratio of the two pathogens based on gender, disease type, complication, past medical history, or smoking history. However, the percentage of patients with M. intracellulare infection was significantly higher among those with underlying lung disease than among those without lung disease (p = 0.0217). The percentage of patients with M. intracellulare infection rose significantly with age (p = 0.0296). This age-related change was more significant in women (p = 0.0018). When district-wise analysis was performed for Japan, the percentage of M. intracellulare infection was higher in the Chugoku/Shikoku and Kyushu districts whereas the percentage of M. avium infection was higher in the other districts. This survey revealed some differences in the clinical and epidemiologic features of M. avium and M. intracellulare infection. The significant predominance of M. avium infection among relatively young women is suggestive of an increase in the M. avium/M. intracellulare infection ratio among women in the future. Copyright © 2017 The Japanese Respiratory Society. Published by Elsevier B.V. All rights reserved.

  17. 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....

  18. 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.

  19. The bright-star masks for the HSC-SSP survey

    Science.gov (United States)

    Coupon, Jean; Czakon, Nicole; Bosch, James; Komiyama, Yutaka; Medezinski, Elinor; Miyazaki, Satoshi; Oguri, Masamune

    2018-01-01

    We present the procedure to build and validate the bright-star masks for the Hyper-Suprime-Cam Strategic Subaru Proposal (HSC-SSP) survey. To identify and mask the saturated stars in the full HSC-SSP footprint, we rely on the Gaia and Tycho-2 star catalogues. We first assemble a pure star catalogue down to GGaia Digital Sky Survey (SDSS). We perform visual inspection on the early data from the S16A internal release of HSC-SSP, finding that our star catalogue is 99.2% pure down to GGaia < 18. Second, we build the mask regions in an automated way using stacked detected source measurements around bright stars binned per GGaia magnitude. Finally, we validate those masks by visual inspection and comparison with the literature of galaxy number counts and angular two-point correlation functions. This version (Arcturus) supersedes the previous version (Sirius) used in the S16A internal and DR1 public releases. We publicly release the full masks and tools to flag objects in the entire footprint of the planned HSC-SSP observations at "ftp://obsftp.unige.ch/pub/coupon/brightStarMasks/HSC-SSP/".

  20. Searching for moving objects in HSC-SSP: Pipeline and preliminary results

    Science.gov (United States)

    Chen, Ying-Tung; Lin, Hsing-Wen; Alexandersen, Mike; Lehner, Matthew J.; Wang, Shiang-Yu; Wang, Jen-Hung; Yoshida, Fumi; Komiyama, Yutaka; Miyazaki, Satoshi

    2018-01-01

    The Hyper Suprime-Cam Subaru Strategic Program (HSC-SSP) is currently the deepest wide-field survey in progress. The 8.2 m aperture of the Subaru telescope is very powerful in detecting faint/small moving objects, including near-Earth objects, asteroids, centaurs and Tran-Neptunian objects (TNOs). However, the cadence and dithering pattern of the HSC-SSP are not designed for detecting moving objects, making it difficult to do so systematically. In this paper, we introduce a new pipeline for detecting moving objects (specifically TNOs) in a non-dedicated survey. The HSC-SSP catalogs are sliced into HEALPix partitions. Then, the stationary detections and false positives are removed with a machine-learning algorithm to produce a list of moving object candidates. An orbit linking algorithm and visual inspections are executed to generate the final list of detected TNOs. The preliminary results of a search for TNOs using this new pipeline on data from the first HSC-SSP data release (2014 March to 2015 November) present 231 TNO/Centaurs candidates. The bright candidates with Hr 5 show that the best-fitting slope of a single power law to absolute magnitude distribution is 0.77. The g - r color distribution of hot HSC-SSP TNOs indicates a bluer peak at g - r = 0.9, which is consistent with the bluer peak of the bimodal color distribution in literature.

  1. Some aspects of cultivation and utilization of waxy maize (Zea mays L. ssp. ceratina

    Directory of Open Access Journals (Sweden)

    Agnieszka Klimek-Kopyra

    2012-10-01

    Full Text Available This paper is a review of available literature on Zea mays L.ssp.ceratina. It contains information on the origin, cul- tivation and utilization of waxy maize in the world and can be a contribution to the development of new research on maize cultivation and starch processing technology. Maize, as an old and economically important cereal, played an enormous role in the ancient civilisations of the New World. Among the maize subspecies compared, Z. mays ssp. indurata and Z. mays ssp. indentata are now the most important in Poland. The subspecies Z. mays ssp. saccharata has a marginal role, while Z. mays ssp. ceratina has not been hitherto cultivated. Decisions to introduce the subspecies Z. mays ssp. ceratina into cultivation are based on different grounds, taking into account both agro-climatic conditions and industrial uses of grain processing products. The growing demand for maize grain, stimulated by the increased demand for maize starch and oil in the global market as raw materials that are important in food production, is an impulse for the development of agrobiological research. The development of the starch industry, associated with the demand for industrial starch, will probably contribute to increased interest in this subspecies in Central Europe, also including Poland. Waxy maize grain can be a major ingredient of high-energy feeds for livestock, replacing in this role the type of maize that has been grown for this purpose until now. A great advantage of waxy maize is its specific structure of starch, due to its unique and high amylopectin content (95-98%, which creates unlimited possibilities of industrial use. Currently, waxy maize acreage in Europe does not exceed 2% of the maize crop area in this continent.

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

    Science.gov (United States)

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

    2016-09-01

    Diseases which are caused by non-tuberculous mycobacteria (NTM) are an increasing problem in the developed countries. In Latvia, one of the most clinically important members of NTM is Mycobacterium avium (M. avium), an opportunistic pathogen which has been isolated from several lung disease patients and tissue samples of slaughter pigs. This study was designed to characterize the genetic diversity of the M. avium isolates in Latvia and to compare the distribution of genotypic patterns among humans and pigs. Eleven (Hall and Salipante, 2010) clinical M. avium samples, isolated from patients of Center of Tuberculosis and Lung Diseases (years 2003-2010), and 32 isolates from pig necrotic mesenterial lymph nodes in different regions (years 2003-2007) were analyzed. The majority (42 of 43) of samples were identified as M. avium subsp. hominissuis; one porcine isolate belonged to M. avium subsp. avium. MIRU-VNTR genotyping revealed 13 distinct genotypes, among which nine genotype patterns, including M. avium subsp. avium isolate, were newly identified. IS1245 RFLP fingerprinting of 25 M. avium subsp. hominissuis samples yielded 17 different IS1245 RFLP patterns, allowing an efficient discrimination of isolates. Clusters of identical RFLP profiles were observed within host species, geographical locations and time frame of several years. Additional in silico analysis on simulated MIRU-VNTR genotype population datasets showed that the MIRU-VNTR pattern similarity could partly arise due to probabilistic increase of acquiring homoplasy among subpopulations, thus the similar MIRU-VNTR profiles of M. avium strains even in close geographical proximity should be interpreted with caution. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Variable-Number Tandem-Repeat Analysis of Respiratory and Household Water Biofilm Isolates of “Mycobacterium avium subsp. hominissuis” with Establishment of a PCR Database

    Science.gov (United States)

    Iakhiaeva, Elena; Howard, Susan T.; Brown Elliott, Barbara A.; McNulty, Steven; Newman, Kristopher L.; Falkinham, Joseph O.; Williams, Myra; Kwait, Rebecca; Lande, Leah; Vasireddy, Ravikiran; Turenne, Christine

    2016-01-01

    “Mycobacterium avium subsp. hominissuis” is an important cause of pulmonary disease. It is acquired from environmental sources, but there is no methodology for large population studies. We evaluated the potential of variable-number tandem-repeat (VNTR) analysis. Clinical and household biofilm M. avium isolates underwent molecular identification. Testing for IS901 was done to separate M. avium subsp. avium from M. avium subsp. hominissuis. VNTR types were defined using VNTR loci, and subtyping was performed using 3′ hsp65 and internal transcribed spacer (ITS) sequencing. Forty-nine VNTR types and eight subtypes of M. avium subsp. hominissuis (IS901 negative) were identified among 416 isolates of M. avium from 121 patients and 80 biofilm sites. Of those types, 67% were found only among patient isolates, 11% only among household water isolates, and 23% among both. Of 13 VNTR types that included ≥4 patients, the majority (61.5%) represented geographic clustering (same city). Most VNTR types with multiple patients belonged to the same 3′ hsp65 sequence code (sequevar). A total of 44 isolates belonging to four M. avium subsp. hominissuis VNTR types (8%), including three with the rare Mav-F ITS sequence and 0/8 subspecies, produced amplicons with IS901 PCR primers. By sequencing, all 44 amplicons were not IS901 but ISMav6, which was recently observed in Japan but had not been previously described among U.S. isolates. VNTR analysis of M. avium subsp. hominissuis isolates is easier and faster than pulsed-field gel electrophoresis. Seven VNTR loci separated 417 isolates into 49 types. No isolates of M. avium subsp. avium were identified. The distributions of the VNTR copy numbers, the allelic diversity, and the low prevalence of ISMav6 differed from the findings for respiratory isolates reported from Japan. PMID:26739155

  4. Radiometric assessment of the sensitivity to antituberculotics of Mycobacterium avium-intracellulare and Mycobacterium xenopi

    International Nuclear Information System (INIS)

    Kubin, M.; Lindholm-Levy, P.; Heifets, L. B.

    1994-01-01

    The macrodilution radiometric method using Middlebrook's 7H12 liquid medium enriched with 14 C-palmitic acid, where the growth activity is monitored by measuring liberated 14 CO 2 , was applied to 25 strains of the Mycobacterium avium complex and to 20 strains of Mycobacterium xenopi to determine the minimal inhibitory concentrations of the following chemotherapeutical agents: ciprofloxacine, clofazimine, rifampin, cycloserine, kanamycin, etionamide, ethambutol, and amikacin. In the case of the M. avium complex, slightly or completely resistant strains were found for the majority of drugs. The sensitive strain proportion was highest with clofazimine and amikacin. The M. xenopis strains exhibited generally lower minimal inhibitory concentrations than the avian mycobacteria for all drugs except for cycloserine and ethambutol. The radiometric method using the BACTEC system was found suitable for the determination of the sensitivity of mycobacteria to chemotherapeutic agents: the results are obtained rapidly, within 8 days following inoculation, and the minimal inhibitory concentrations can be evaluated quantitatively. 1 tab., 8 refs

  5. 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.

  6. Microaerobic growth and anaerobic survival of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum

    Directory of Open Access Journals (Sweden)

    Amy Herndon Lewis

    2015-01-01

    Full Text Available Representative strains of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum (MAIS grew at equal rates in laboratory medium at 21% (air and 12% oxygen. Growth in 6% oxygen proceeded at a 1.4–1.8-fold lower rate. Colony formation was the same at 21% (air and 6% oxygen. The MAIS strains survived rapid shifts from aerobic to anaerobic conditions as measured by two experimental approaches (Falkinham (1996 [1]. MAIS cells grown aerobically to log phase in broth were diluted, spread on agar medium, and incubated anaerobically for up to 20 days at 37 °C. Although no colonies formed anaerobically, upon transfer to aerobic conditions, greater than 25% of the colony forming units (CFU survived after 20 days of anaerobic incubation (Prince et al. (1989 [2]. MAIS cells grown in broth aerobically to log phase were sealed and vigorous agitation led to oxygen depletion (Wayne model. After 12 days anaerobic incubation, M. avium and M. scrofulaceum survival were high (>50%, while M. intracellulare survival was lower (22%. M. avium cells shifted to anaerobiosis in broth had increased levels of glycine dehydrogenase and isocitrate lyase. Growth of MAIS strains at low oxygen levels and their survival following a rapid shift to anaerobiosis is consistent with their presence in environments with fluctuating oxygen levels.

  7. Microaerobic growth and anaerobic survival of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum.

    Science.gov (United States)

    Lewis, Amy Herndon; Falkinham, Joseph O

    2015-03-01

    Representative strains of Mycobacterium avium, Mycobacterium intracellulare and Mycobacterium scrofulaceum (MAIS) grew at equal rates in laboratory medium at 21% (air) and 12% oxygen. Growth in 6% oxygen proceeded at a 1.4-1.8-fold lower rate. Colony formation was the same at 21% (air) and 6% oxygen. The MAIS strains survived rapid shifts from aerobic to anaerobic conditions as measured by two experimental approaches (Falkinham (1996) [1]). MAIS cells grown aerobically to log phase in broth were diluted, spread on agar medium, and incubated anaerobically for up to 20 days at 37°C. Although no colonies formed anaerobically, upon transfer to aerobic conditions, greater than 25% of the colony forming units (CFU) survived after 20 days of anaerobic incubation (Prince et al. (1989) [2]). MAIS cells grown in broth aerobically to log phase were sealed and vigorous agitation led to oxygen depletion (Wayne model). After 12 days anaerobic incubation, M. avium and M. scrofulaceum survival were high (>50%), while M. intracellulare survival was lower (22%). M. avium cells shifted to anaerobiosis in broth had increased levels of glycine dehydrogenase and isocitrate lyase. Growth of MAIS strains at low oxygen levels and their survival following a rapid shift to anaerobiosis is consistent with their presence in environments with fluctuating oxygen levels. Copyright © 2015 Asian African Society for Mycobacteriology. Published by Elsevier Ltd. All rights reserved.

  8. [Identification of Mycobacterium avium-intracellulare complex by PCR of AIDS and disseminated mycobacteriosis].

    Science.gov (United States)

    García-Elorriaga, Guadalupe; Degollado-Estrada, Edgar; Villagómez-Ruiz, Alfredo; Cortés-Torres, Nancy; Arreguín-Reséndiz, Lilián; Del Rey-Pineda, Guillermo; González-Bonilla, César

    2016-01-01

    The aim of this study is to differentially identify MAC by PCR in patients with AIDS and disseminated mycobacteriosis. A cross sectional study was conducted in Mexico to identify MAC by Molecular Biology. Two sets of primers were synthesized: MAV and MIN, for M. avium and M. intracellulare, respectively. Whole-cell DNAs obtained from 29 clinical isolates and clinical serum specimens from other 24 patients with AIDS and disseminated mycobacterial infection were extracted and amplified by PCR with the MAV and MIN primers. The MAV and MIN primers each amplified one highly specific 1.3-kb segment of the homologous DNA, respectively. Twenty-nine DNAs from MAC clinical isolates identified by Gen-Probe AccuProbes were amplified with the MAV primers. Of the 24 clinical samples, 3 were positive for M. avium and 6 for M. tuberculosis. Our results demonstrated that PCR technique could be applied for the differentiation of M. avium and M. intracellulare by specific 16S rRNA primers. In patients with advanced stage AIDS and in whom disseminated mycobacteriosis is suspected, the presence of anemia (even with negative cultures), elevated alkaline phosphatase and a median CD4 count of 15.9/mL, the diagnosis of infection by MAC should be strongly considered; we suggest that in accordance with our findings, a more precise stratification of patients in terms of their CD4 T cell counts is warranted.

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

    Science.gov (United States)

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

    2009-04-01

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

  10. L. peruvianum as a source for resistance to C michiganensis ssp. michiganensis in tomato

    NARCIS (Netherlands)

    Heusden, van S.; Vrielink, R.; Lindhout, P.

    1995-01-01

    Twenty three accessions of L. peruvianum were screened for resistance to Clavibater michiganensis ssp. michiganensis (bacterial canker). Resistance was detected in five accessions. One of these, L. peruvianum LA2157 was crossed with a susceptible accession of L. peruvainum and with the cultivated

  11. Identification of functional candidates amongst hypothetical proteins of Treponema pallidum ssp. pallidum.

    Science.gov (United States)

    Naqvi, Ahmad Abu Turab; Shahbaaz, Mohd; Ahmad, Faizan; Hassan, Md Imtaiyaz

    2015-01-01

    Syphilis is a globally occurring venereal disease, and its infection is propagated through sexual contact. The causative agent of syphilis, Treponema pallidum ssp. pallidum, a Gram-negative sphirochaete, is an obligate human parasite. Genome of T. pallidum ssp. pallidum SS14 strain (RefSeq NC_010741.1) encodes 1,027 proteins, of which 444 proteins are known as hypothetical proteins (HPs), i.e., proteins of unknown functions. Here, we performed functional annotation of HPs of T. pallidum ssp. pallidum using various database, domain architecture predictors, protein function annotators and clustering tools. We have analyzed the sequences of 444 HPs of T. pallidum ssp. pallidum and subsequently predicted the function of 207 HPs with a high level of confidence. However, functions of 237 HPs are predicted with less accuracy. We found various enzymes, transporters, binding proteins in the annotated group of HPs that may be possible molecular targets, facilitating for the survival of pathogen. Our comprehensive analysis helps to understand the mechanism of pathogenesis to provide many novel potential therapeutic interventions.

  12. Proteomic analysis of bronchoalveolar lavage fluid proteins from mice infected with Francisella tularensis ssp. novicida.

    Science.gov (United States)

    Varnum, Susan M; Webb-Robertson, Bobbie-Jo M; Pounds, Joel G; Moore, Ronald J; Smith, Richard D; Frevert, Charles W; Skerrett, Shawn J; Wunschel, David

    2012-07-06

    Francisella tularensis causes the zoonosis tularemia in humans and is one of the most virulent bacterial pathogens. We utilized a global proteomic approach to characterize protein changes in bronchoalveolar lavage fluid from mice exposed to one of three organisms, F. tularensis ssp. novicida, an avirulent mutant of F. tularensis ssp. novicida (F.t. novicida-ΔmglA), and Pseudomonas aeruginosa. The composition of bronchoalveolar lavage fluid (BALF) proteins was altered following infection, including proteins involved in neutrophil activation, oxidative stress, and inflammatory responses. Components of the innate immune response were induced including the acute phase response and the complement system; however, the timing of their induction varied. F. tularensis ssp. novicida infected mice do not appear to have an effective innate immune response in the first hours of infection; however, within 24 h, they show an upregulation of innate immune response proteins. This delayed response is in contrast to P. aeruginosa infected animals which show an early innate immune response. Likewise, F.t. novicida-ΔmglA infection initiates an early innate immune response; however, this response is diminished by 24 h. Finally, this study identifies several candidate biomarkers, including Chitinase 3-like-1 (CHI3L1 or YKL-40) and peroxiredoxin 1, that are associated with F. tularensis ssp. novicida but not P. aeruginosa infection.

  13. Essential oil composition of Valeriana officinalis ssp collina cultivated in Bulgaria

    NARCIS (Netherlands)

    Bos, R; Hendriks, H; Pras, N; St Stojanova, A; Georgiev, EV

    2000-01-01

    The essential oils obtained from air-dried subterranean parts of Valeriana officinalis ssp. collina (Wallr.) Nyman (Valerianaceae), 'Shipka,' was analyzed by GC and GC/MS, and 62 components were identified. The oil yield, collected in plants grown in 1995 and 1938, was 0.42% (v/m) and 0.40% (v/m)

  14. PENGEMBANGAN SSP FISIKA BERBASIS PENDEKATAN CTL UNTUK MENINGKATKAN KETERAMPILAN PROSES SAINS DAN MOTIVASI BELAJAR

    Directory of Open Access Journals (Sweden)

    Faiq Makhdum Noor

    2015-04-01

    Full Text Available This research aims to develop a learning device as Subject-Specific Pedagogy (SSP of physics, and to improve science process skills and learning motivation of the students. This research used the 4D model. The subjects of this research were the XI grade students of Muhammadiyah High School 2 Yogyakarta. The data collecting instruments were expert validation sheets, science process skill observation sheets, questionnaire of students’ response to the student books and worksheets, question-naire of students’ response to teaching process, students’ motivation learning sheets, and science process skill paper test. The research data were collected through observation, questionnaire, and paper test, and analyzed quantitatively. The results show as follows. (1 The development of physics SSP is carried out in accordance with the procedures of 4D model product development. (2 The results of expert’s validation show the physics SSP is in the very good criteria. (3 The readability of the student books and worksheets is in the good criteria. (4 The increasing of the score of science process skills and students’ learning motivation in the experimental class is higher than that in the control class. This shows that the physics SSP is fit for use in physics teaching.

  15. Second generation bioethanol production from Saccharum spontaneum L. ssp. aegyptiacum (Willd.) Hack.

    Science.gov (United States)

    Danilo Scordia; Salvatore L. Consentino; Thomas W. Jeffries

    2010-01-01

    Saccharum (Saccharum spontaneum L. ssp. aegyptiacum (Willd.) Hack.), is a rapidly growing, wide ranging high-yield perennial, suitable for second generation bioethanol production. This study evaluated oxalic acid as a pretreatment for bioconversion. Overall sugar yields, sugar degradation products, enzymatic glucan hydrolysis and ethanol production were studied as...

  16. Proteomic analysis of bronchoalveolar lavage fluid proteins from mice infected with Francisella tularensis ssp novicida

    Energy Technology Data Exchange (ETDEWEB)

    Varnum, Susan M.; Webb-Robertson, Bobbie-Jo M.; Pounds, Joel G.; Moore, Ronald J.; Smith, Richard D.; Frevert, Charles; Skerret, Shawn J.; Wunschel, David S.

    2012-07-06

    Francisella tularensis causes the zoonosis tularemia in humans and is one of the most virulent bacterial pathogens. We utilized a global proteomic approach to characterize protein changes in bronchoalveolar lavage fluid from mice exposed to one of three organisms, F. tularensis ssp. novicida, an avirulent mutant of F. tularensis ssp. novicida (F.t. novicida-ΔmglA); and Pseudomonas aeruginosa. The composition of BALF proteins was altered following infection, including proteins involved in neutrophil activation, oxidative stress and inflammatory responses. Components of the innate immune response were induced including the acute phase response and the complement system, however the timing of their induction varied. Francisella tularensis ssp. novicida infected mice do not appear to have an effective innate immune response in the first hours of infection, however within 24 hours they show an upregulation of innate immune response proteins. This delayed response is in contrast to P. aeruginosa infected animals which show an early innate immune response. Likewise, F.t. novicida-ΔmglA infection initiates an early innate immune response, however this response is dimished by 24 hours. Finally, this study identifies several candidate biomarkers, including Chitinase 3-like-1 (CHI3L1 or YKL-40) and peroxiredoxin 1, that are associated with F. tularensis ssp. novicida but not P. aeruginosa infection.

  17. Attempting to restore mountain big sagebrush (Artemisia tridentata ssp. vaseyana) four years after fire

    Science.gov (United States)

    Restoration of shrubs is increasingly needed throughout the world because of altered fire regimes, anthropogenic disturbance, and over-utilization. The native shrub mountain big sagebrush (Artemisia tridentata Nutt. ssp. vaseyana (Rydb.) Beetle) is a restoration priority in western North America be...

  18. THE ISOLATION AND IDENTIFICATION OF MYCOBACTERIUM AVIUM COMPLEX (MAC) RECOVERED FROM LOS ANGELES POTABLE WATER, A POSSIBLE SOURCE OF INFECTION IN AIDS PATIENTS

    Science.gov (United States)

    Los Angeles water was investigated as a possible source of Mycobacterium avium complex (MAC) infection in patients with AIDS. MAC consists of M.avium (MA), M. intracellulare (MI) and Mycobacterium X (MX)(positive for MAC by DNA probe but not MA or MI). The study included 13 reser...

  19. PENGEMBANGAN SSP TEMATIK-INTEGRATIF UNTUK MEMBANGUN KARAKTER DISIPLIN DAN KREATIF SISWA KELAS I SD

    Directory of Open Access Journals (Sweden)

    Wawan Wahyu Setyawan

    2015-01-01

    DEVELOPING THEMATIC-INTEGRATED SSP FOR BUILDING THE CHARACTERS OF DISCIPLINE AND CREATIVE OF GRADE I STUDENT ELEMENTARY SCHOOL ABSTRACT This study aims to produce integrated learning sets in the form of thematic SSP that can develop the characters of discipline and creative of grade I students of elementary school.This was a research and development study consisting of seven steps, namely: preliminary study and data collection, planning, product draft development, preliminary field tryout, revision of the tryout result, field tryout, product finalization based the field tryout result, field implementation testing, final product finalization, and dissemination. The developed SSP was evaluated by a science subject expert to assess the validity. The tryout subjects comprised 82 students consisting of 26 students of SD Baran, Pundong, Bantul, DIY in the small-scale tryout, 25 students in the control class of SD Manding Tengah, Bantul, DIY and 31 students in the experimental class of SD 1 Tulung, Bantul, DIY. The results of this study are in the form of SSP consisting of: a syllabus, lesson plans, course sheets, student worksheets, and evaluation sheets. The results of the evaluation show that the developed syllabus is good score of 47. The developed lesson plans score of 67, student course sheets score of 65, student worksheets score of 47, and assessment instruments, are very good , while the developed learning media are good score of 66. The results of the tryout show that the developed SSP otherwise meet the criteria for the practical views of teacher assessment score of 281 and a score of 730 student responses. In field trials the results obtained for the control class average pretest score of 70.8 and an average posttest score of 80.8. While the experimental class scored an average of 74.1 pretest and posttest score of 90.9. Improved control class scores of 10 and an increase of 16.8 experimental class. Teaching using developed SSP is capable of developing

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

    Directory of Open Access Journals (Sweden)

    José de Arimatéa Freitas

    2001-06-01

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

  1. NCBI nr-aa BLAST: CBRC-CREM-01-1284 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CREM-01-1284 ref|NP_962383.1| SugI [Mycobacterium avium subsp. paratuberculosis... K-10] gb|AAS05999.1| SugI [Mycobacterium avium subsp. paratuberculosis K-10] NP_962383.1 2e-85 55% ...

  2. 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% ...

  3. Streptococcus mutans SpaP binds to RadD of Fusobacterium nucleatum ssp. polymorphum.

    Science.gov (United States)

    Guo, Lihong; Shokeen, Bhumika; He, Xuesong; Shi, Wenyuan; Lux, Renate

    2017-10-01

    Adhesin-mediated bacterial interspecies interactions are important elements in oral biofilm formation. They often occur on a species-specific level, which could determine health or disease association of a biofilm community. Among the key players involved in these processes are the ubiquitous fusobacteria that have been recognized for their ability to interact with numerous different binding partners. Fusobacterial interactions with Streptococcus mutans, an important oral cariogenic pathogen, have previously been described but most studies focused on binding to non-mutans streptococci and specific cognate adhesin pairs remain to be identified. Here, we demonstrated differential binding of oral fusobacteria to S. mutans. Screening of existing mutant derivatives indicated SpaP as the major S. mutans adhesin specific for binding to Fusobacterium nucleatum ssp. polymorphum but none of the other oral fusobacteria tested. We inactivated RadD, a known adhesin of F. nucleatum ssp. nucleatum for interaction with a number of gram-positive species, in F. nucleatum ssp. polymorphum and used a Lactococcus lactis heterologous SpaP expression system to demonstrate SpaP interaction with RadD of F. nucleatum ssp. polymorphum. This is a novel function for SpaP, which has mainly been characterized as an adhesin for binding to host proteins including salivary glycoproteins. In conclusion, we describe an additional role for SpaP as adhesin in interspecies adherence with RadD-SpaP as the interacting adhesin pair for binding between S. mutans and F. nucleatum ssp. polymorphum. Furthermore, S. mutans attachment to oral fusobacteria appears to involve species- and subspecies-dependent adhesin interactions. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

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

    Directory of Open Access Journals (Sweden)

    Robson F.C. Almeida

    2006-12-01

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

  5. Chemical Composition of Different Botanical Origin Honeys Produced by Sicilian Black Honeybees (Apis mellifera ssp. sicula).

    Science.gov (United States)

    Mannina, Luisa; Sobolev, Anatoly P; Di Lorenzo, Arianna; Vista, Silvia; Tenore, Gian Carlo; Daglia, Maria

    2015-07-01

    In 2008 a Slow Food Presidium was launched in Sicily (Italy) for an early warning of the risk of extinction of the Sicilian native breed of black honeybee (Apis mellifera L. ssp sicula). Today, the honey produced by these honeybees is the only Sicilian honey produced entirely by the black honeybees. In view of few available data regarding the chemical composition of A. mellifera ssp. sicula honeys, in the present investigation the chemical compositions of sulla honey (Hedysarum coronarium L.) and dill honey (Anethum graveolens L.) were studied with a multimethodological approach, which consists of HPLC-PDA-ESI-MSn and NMR spectroscopy. Moreover, three unifloral honeys (lemon honey (obtained from Citrus limon (L.) Osbeck), orange honey (Citrus arantium L.), and medlar honey (Eriobotrya japonica (Thunb.) Lindl)), with known phenol and polyphenol compositions, were studied with NMR spectroscopy to deepen the knowledge about sugar and amino acid compositions.

  6. ARE ENTEROCOCCUS FAECIUM CRL 183 AND LACTOBACILLUS HELVETICUS SSP. JUGURTI 416 ABLE TO PRODUCE ANTIMICROBIAL SUBSTANCE?

    Directory of Open Access Journals (Sweden)

    R. BEDANI

    2008-12-01

    Full Text Available

    Production of antimicrobial substance by E. faecium CRL 183 and L. helveticus ssp jugurti 416 was tested against pathogenic bacteria and microorganisms isolated from Wistar rat faeces, using a spot-on-the-lawn assay. Three pathogenic bacteria (Listeria monocytogenes 1/2a, Salmonella enteridites 193/95, Escherichia coli O157:H7 and twenty-fi ve colonies, isolated from animal faeces, belonging to the following groups Enterococcus, Lactobacillus, Clostridium, Bacteroides and Enterobacteriaceae, were used as indicator microorganisms. The results showed that E. faecium CRL 183 and L. helveticus ssp jugurti 416 did not produce signifi cant antimicrobial activity against the indicator microorganisms tested. More tests needed to be carried out with other indicator bacteria and other culture media to confi rm the lack of antimicrobial production.

  7. Chemical Composition and Antimicrobial Activity of Thymus praecox Opiz ssp. polytrichus Essential Oil from Serbia

    Directory of Open Access Journals (Sweden)

    Nada V. Petrović

    2016-03-01

    Full Text Available Chemical composition and antimicrobial activity of the essential oil of wild growing Thymus praecox Opiz ssp. polytrichus were studied. trans-Nerolidol (19.79%, germacrene D (18.48% and thymol (9.62% were the main components in essential oil. This study is the first report of the antimicrobial activity of essential oil obtained from the T. praecox Opiz ssp. polytrichus. Antimicrobial activity of essential oil was investigated on Bacillus cereus, Micrococcus flavus, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, Pseudomonas aeruginosa, Enterobacter cloacae, Salmonella typhimurium, Aspergillus fumigatus, A. versicolor, A. ochraceus, A. niger, Trichoderma viride, Penicillium funiculosum, P. ochrochloron, and P. verrucosum var. cyclopium strains. In the antimicrobial assays, essential oil showed high antimicrobial potential (MIC 19–150 m g/mL, MBC 39–300 m g/mL for bacteria; and MIC 19.5–39 m g/mL, MFC 39–78 m g/mL for fungi.

  8. Molecular mechanisms regulating flowering time in sweet cherry (Prunus avium L.)

    DEFF Research Database (Denmark)

    Ionescu, Irina Alexandra

    as a result of hydrogen cyanamide treatment: the jasmonate pathway, the hydrogen cyanide pathway and the cytokinin pathway. We further analyzed the levels of cyanogenic glucosides and their derivatives during endodormancy and its release in sweet cherry and almond (Prunus dulcis (Mill.) D. A. Webb). Prunasin...... example is the agrochemical hydrogen cyanamide, which besides its successful application in agriculture constitutes an excellent experimental system to research controlled endodormancy release. In this project, we treated dormant sweet cherry (Prunus avium L.) flower buds with hydrogen cyanamide...

  9. 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.

  10. Henoch-Schönlein purpura associated with pulmonary Mycobacterium avium-intracellulare complex.

    Science.gov (United States)

    Yano, Shuichi

    2004-09-01

    A 57-year-old woman presented with bloody sputum and high grade fever. She had been treated for Mycobacterium avium-intracellulare complex (MAC). High grade fever slightly decreased and bloody sputum disappeared after two weeks, but low grade fever persisted. After 3 days of recurrence of bloody sputum, she suddenly complained of palpable pururitic lesions on the bilateral lower extremities with bilateral gonalgia. Although there are some reports of direct skin lesions due to MAC, there are no reports of hypersensitivity vasculitis, Henoch-Schönlein purpura, in MAC. It is necessary to consider MAC infection as a potential cause of Henoch-Schönlein purpura.

  11. 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...... 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...

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

    International Nuclear Information System (INIS)

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

    1988-01-01

    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

  13. The 172-kb genomic DNA region of the O. rufipogon yld1.1 locus: comparative sequence analysis with O. sativa ssp. japonica and O. sativa ssp. indica.

    Science.gov (United States)

    Song, Beng-Kah; Hein, Ingo; Druka, Arnis; Waugh, Robbie; Marshall, David; Nadarajah, Kalaivani; Yap, Soon-Joo; Ratnam, Wickneswari

    2009-02-01

    Common wild rice (Oryza rufipogon) plays an important role by contributing to modern rice breeding. In this paper, we report the sequence and analysis of a 172-kb genomic DNA region of wild rice around the RM5 locus, which is associated with the yield QTL yld1.1. Comparative sequence analysis between orthologous RM5 regions from Oryza sativa ssp. japonica, O. sativa ssp. indica and O. rufipogon revealed a high level of conserved synteny in the content, homology, structure, orientation, and physical distance of all 14 predicted genes. Twelve of the putative genes were supported by matches to proteins with known function, whereas two were predicted by homology to rice and other plant expressed sequence tags or complementary DNAs. The remarkably high level of conservation found in coding, intronic and intergenic regions may indicate high evolutionary selection on the RM5 region. Although our analysis has not defined which gene(s) determine the yld1.1 phenotype, allelic variation and the insertion of transposable elements, among other nucleotide changes, represent potential variation responsible for the yield QTL. However, as suggested previously, two putative receptor-like protein kinase genes remain the key suspects for yld1.1.

  14. Consumption of Dairy Yogurt Containing Lactobacillus paracasei ssp. paracasei, Bifidobacterium animalis ssp. lactis and Heat-Treated Lactobacillus plantarum Improves Immune Function Including Natural Killer Cell Activity

    Directory of Open Access Journals (Sweden)

    Ayoung Lee

    2017-05-01

    Full Text Available The aim of this study was to investigate the impact of consuming dairy yogurt containing Lactobacillus paracasei ssp. paracasei (L. paracasei, Bifidobacterium animalis ssp. lactis (B. lactis and heat-treated Lactobacillus plantarum (L. plantarum on immune function. A randomized, open-label, placebo-controlled study was conducted on 200 nondiabetic subjects. Over a twelve-week period, the test group consumed dairy yogurt containing probiotics each day, whereas the placebo group consumed milk. Natural killer (NK cell activity, interleukin (IL-12 and immunoglobulin (Ig G1 levels were significantly increased in the test group at twelve weeks compared to baseline. Additionally, the test group had significantly greater increases in serum NK cell activity and interferon (IFN-γ and IgG1 than placebo group. Daily consumption of dairy yogurt containing L. paracasei, B. lactis and heat-treated L. plantarum could be an effective option to improve immune function by enhancing NK cell function and IFN-γ concentration (ClinicalTrials.gov: NCT03051425.

  15. Evaluating the Strange Situation Procedure (SSP to assess the bond between dogs and humans.

    Directory of Open Access Journals (Sweden)

    Therese Rehn

    Full Text Available The Strange Situation Procedure (SSP is increasingly being used to study attachment between dogs and humans. It has been developed from the Ainsworth Strange Situation Procedure, which is used extensively to investigate attachment between children and their parents. In this experiment, 12 female beagle dogs were tested in two treatments to identify possible order effects in the test, a potential weakness in the SSP. In one treatment (FS, dogs participated together with a 'familiar person' and a 'stranger'. In a control treatment (SS, the same dogs participated together with two unfamiliar people, 'stranger A' and 'stranger B'. Comparisons were made between episodes within as well as between treatments. As predicted in FS, dogs explored more in the presence of the familiar person than the stranger. Importantly, they also explored more in the presence of stranger A (who appeared in the same order as the familiar person and followed the same procedure than stranger B in SS. Furthermore, comparisons between treatments, where a familiar person was present in FS and stranger A was present in SS, showed no differences in exploration. In combination, these results indicate that the effect of a familiar person on dogs' exploratory behaviour, a key feature when assessing secure attachment styles, could not be tested reliably due to the order in which the familiar person and the stranger appear. It is proposed that in the future only counterbalanced versions of the SSP are used. Alternatively, since dogs reliably initiated more contact with the familiar person compared to the strangers, it is suggested that future studies on attachment in dogs towards humans should focus either on the behaviour of the dog in those episodes of the SSP when the person returns, or on reunion behaviour in other studies, specially designed to address dog-human interactions at this time.

  16. Failure of PCR to Detect Treponema pallidum ssp. pertenue DNA in Blood in Latent Yaws.

    Science.gov (United States)

    Marks, Michael; Katz, Samantha; Chi, Kai-Hua; Vahi, Ventis; Sun, Yongcheng; Mabey, David C; Solomon, Anthony W; Chen, Cheng Y; Pillay, Allan

    2015-01-01

    Yaws, caused by Treponema pallidum ssp. pertenue, is a neglected tropical disease closely related to venereal syphilis and is targeted for eradication by 2020. Latent yaws represents a diagnostic challenge, and current tools cannot adequately distinguish between individuals with true latent infection and individuals who are serofast following successful treatment. PCR on blood has previously been shown to detect T. pallidum DNA in patients with syphilis, suggesting that this approach may be of value in yaws. We performed real-time PCR for Treponema pallidum ssp. pertenue on blood samples from 140 children with positive T. pallidum Particle Agglutination (TPPA) and Rapid Plasma Reagin (RPR) tests and 7 controls (negative serology), all collected as part of a prospective study of yaws in the Solomon Islands. All samples were also tested by a nested PCR for T. pallidum. 12 patients had clinical evidence of active yaws whilst 128 were considered to have latent yaws. 43 children had high titre rapid plasma reagins (RPRs) of ≥1:32. PCR testing with both assays gave negative results in all cases. It is possible that the failure to detect T. pallidum ssp. pertenue in blood reflects lower loads of organism in latent yaws compared to those in latent infection with T. pallidum ssp. pertenue, and/or a lower propensity for haematogenous dissemination in yaws than in syphilis. As the goal of the yaws control programme is eradication, a tool that can differentiate true latent infection from individuals who are serofast would be of value; however, PCR of blood is not that tool.

  17. Brain SPECT analysis by 3D-SSP and clinical features of Parkinson's disease

    International Nuclear Information System (INIS)

    Mito, Yasunori; Yoshida, Kazuto; Makino, Kenichi; Yabe, Ichiro; Kikuchi, Seiji; Sasaki, Hidenao; Tashiro, Kunio

    2006-01-01

    The aim of the present study is to investigate the association of symptoms in Parkinson's disease (PD) with cerebral perfusion on single photon emission computed tomography (SPECT). The clinical features of PD were compared with SPECT images of the brain obtained by three-dimensional stereotactic surface projection (3D-SSP) analysis. Thirty-eight patients who had PD without dementia (17 men and 21 women with a mean age of 68.6±4.7 years) were enrolled in this study. Their symptoms were rated using the unified parkinson's disease rating scale (UPDRS). Within a week, all patients were examined by SPECT with I-123, and reconstructed images were analyzed with 3D-SSP using an image-analysis software, iSSP ver. 3.5. Data on brain surface perfusion extracted by 3D-SSP analysis were compared between the PD patients and the normal control group. The same comparisons were made for subgroups of PD patients with severe symptoms, such as tremor, gait disturbance, bradykinesia, and the UPDRS motor score. Cerebral perfusion was decreased at the anterior cingulate cortex and occipital lobe of the PD patients compared with the normal controls. In the subgroups with severe gait disturbance and severe bradykinesia, additional hypoperfusion was seen at the lateral frontal association and lateral temporal association and the medial frontal gyrus, and by the pixel-by-pixel comparison, perfusion was significantly decreased (p<0.05) at the medial frontal gyrus and anterior cingulate cortex compared with the normal control group. In PD patients, severe gait disturbance and bradykinesia may be correlated with hypoperfusion of the medial aspect of the frontal lobe. This suggests that functional disturbance of the supplementary motor area and other parts of the frontal lobe are involved in the development of gait disturbance and bradykinesia in PD. (author)

  18. Evaluating physiological stress in Sumatran tigers (Panthera tigris ssp. sumatrae) managed in Australian zoos

    OpenAIRE

    Parnell, Tempe; Narayan, Edward J.; Magrath, Michael J. L.; Roe, Sheila; Clark, Giles; Nicolson, Vere; Martin-Vegue, Patrick; Mucci, Al; Hero, Jean-Marc

    2014-01-01

    Glucocorticoid quantification using non-invasive methods provides a powerful tool for assessing the health and welfare of wildlife in zoo-based programmes. In this study, we provide baseline data on faecal-based glucocorticoid (cortisol) monitoring of Sumatran tigers (Panthera tigris ssp. sumatrae) managed at the Melbourne Zoo in Victoria, Australia. We sampled five tigers daily for 60?days. Faecal cortisol metabolites (FCMs) in tiger faecal extracts were quantified using enzyme immunoassays ...

  19. Evaluating the Strange Situation Procedure (SSP) to Assess the Bond between Dogs and Humans

    Science.gov (United States)

    Rehn, Therese; McGowan, Ragen T. S.; Keeling, Linda J.

    2013-01-01

    The Strange Situation Procedure (SSP) is increasingly being used to study attachment between dogs and humans. It has been developed from the Ainsworth Strange Situation Procedure, which is used extensively to investigate attachment between children and their parents. In this experiment, 12 female beagle dogs were tested in two treatments to identify possible order effects in the test, a potential weakness in the SSP. In one treatment (FS), dogs participated together with a ‘familiar person’ and a ‘stranger’. In a control treatment (SS), the same dogs participated together with two unfamiliar people, ‘stranger A’ and ‘stranger B’. Comparisons were made between episodes within as well as between treatments. As predicted in FS, dogs explored more in the presence of the familiar person than the stranger. Importantly, they also explored more in the presence of stranger A (who appeared in the same order as the familiar person and followed the same procedure) than stranger B in SS. Furthermore, comparisons between treatments, where a familiar person was present in FS and stranger A was present in SS, showed no differences in exploration. In combination, these results indicate that the effect of a familiar person on dogs' exploratory behaviour, a key feature when assessing secure attachment styles, could not be tested reliably due to the order in which the familiar person and the stranger appear. It is proposed that in the future only counterbalanced versions of the SSP are used. Alternatively, since dogs reliably initiated more contact with the familiar person compared to the strangers, it is suggested that future studies on attachment in dogs towards humans should focus either on the behaviour of the dog in those episodes of the SSP when the person returns, or on reunion behaviour in other studies, specially designed to address dog-human interactions at this time. PMID:23437277

  20. Stem rust (Puccinia graminis ssp. graminicola Urban its hosts and harmfulness in grasses grown for seed

    Directory of Open Access Journals (Sweden)

    Maria Prończuk

    2013-12-01

    Full Text Available Stem rust development on four species of grasses was studied in field experiments conducted at Radzików in 1997-2001. Population of Puccinia graminis ssp. graminicola from different hosts was characterised and their harmfulness for grass grown for seed was estimated. The materials for study were ecotypes and strains of Lolium perenne, Festuca rubra, Poa pratensis and Deschampsia caespitosa collected in breeding nursery and cultivars and strains of L.perenne, F.rubra, P.pratensis cultivated for seed. It was found that the changes in environmental conditions during last years influenced earlier occurrence of stem rust on grasses in Poland. All examined species were the host of P.graminis ssp. graminicola, however the period of infection of particular hosts were different. L.perenne and D.caespitosa were infected in early summer but F.rubra and P.pratensis in late summer or in the autumn. Morphological analysis of spores of P.graminis ssp. graminicola have shoved significant differences between populations obtained from L.perenne and D.caespitosa. Some differences were found between populations from F.rubra and P.pratensis also, but they need more study. Every year occurrence of stem rust on L.perenne and D.caespitosa and its relation with spring temperature in Radzików indicated that populations of patogen could overwinter in local turf. Incidental appearance of stem rust on F.rubra and P.pratensis in centre of Poland allowed to suppose that spores of these forms might be transfer by wind from other regions. The investigation revealed that stem rust can be dangerous for L.perenne grown for seed when infection occurs at flowering time. It has been established that infection of F.rubra and P.pratensis in autumn should not be disregarded. Damages of leaves by P.graminis ssp. graminicola substantially limited plant heading in the next year.

  1. Extinction risk assessment for the species survival plan (SSP) population of the Bali mynah (Leucopsar rothschildi).

    Science.gov (United States)

    Earnhardt, Joanne M; Thompson, Steven D; Faust, Lisa J

    2009-05-01

    The Bali mynah Species Survival Plan (SSP), an Association of Zoos and Aquariums program, strives to maintain the genetic and demographic health of its population, avoid unplanned changes in size, and minimize the risk of population extinction. The SSP population meets current demographic and genetic objectives with a population size of 209 birds at 61 institutions and 96% genetic diversity (GD) retained from the source population. However, participating institutions have expressed concerns regarding space allocation, target population size (TPS), breeding restrictions, inbreeding depression, and harvest in relation to future population availability and viability. Based on these factors, we assess five questions with a quantitative risk assessment, specifically a population viability analysis (PVA) using ZooRisk software. Using an individual-based stochastic model, we project potential population changes under different conditions (e.g. changes in TPS and genetic management) to identify the most effective management actions. Our projections indicate that under current management conditions, population decline and extinction are unlikely and that although GD will decline over 100 years the projected loss does not exceed levels acceptable to population managers (less than 90% GD retained). Model simulations indicate that the combination of two genetic management strategies (i.e. priority breeding based on mean kinship and inbreeding avoidance) benefits the retention of GD and reduces the accumulation of inbreeding. The current TPS (250) is greater than necessary to minimize the risk of extinction for the SSP population but any reduction in TPS must be accompanied by continued application of genetic management. If carefully planned, birds can be harvested for transfer to Bali for a reintroduction program without jeopardizing the SSP population.

  2. Failure of PCR to Detect Treponema pallidum ssp. pertenue DNA in Blood in Latent Yaws.

    Directory of Open Access Journals (Sweden)

    Michael Marks

    Full Text Available Yaws, caused by Treponema pallidum ssp. pertenue, is a neglected tropical disease closely related to venereal syphilis and is targeted for eradication by 2020. Latent yaws represents a diagnostic challenge, and current tools cannot adequately distinguish between individuals with true latent infection and individuals who are serofast following successful treatment. PCR on blood has previously been shown to detect T. pallidum DNA in patients with syphilis, suggesting that this approach may be of value in yaws. We performed real-time PCR for Treponema pallidum ssp. pertenue on blood samples from 140 children with positive T. pallidum Particle Agglutination (TPPA and Rapid Plasma Reagin (RPR tests and 7 controls (negative serology, all collected as part of a prospective study of yaws in the Solomon Islands. All samples were also tested by a nested PCR for T. pallidum. 12 patients had clinical evidence of active yaws whilst 128 were considered to have latent yaws. 43 children had high titre rapid plasma reagins (RPRs of ≥1:32. PCR testing with both assays gave negative results in all cases. It is possible that the failure to detect T. pallidum ssp. pertenue in blood reflects lower loads of organism in latent yaws compared to those in latent infection with T. pallidum ssp. pertenue, and/or a lower propensity for haematogenous dissemination in yaws than in syphilis. As the goal of the yaws control programme is eradication, a tool that can differentiate true latent infection from individuals who are serofast would be of value; however, PCR of blood is not that tool.

  3. Cytogenetic diversity of simple sequences repeats in morphotypes of Brassica rapa ssp. chinensis

    Directory of Open Access Journals (Sweden)

    Jinshuang Zheng

    2016-07-01

    Full Text Available A significant fraction of the nuclear DNA of all eukaryotes is occupied by simple sequence repeats (SSRs. Although thesis sequences have sparked great interest as a means of studying genetic variation, linkage mapping and evolution, little attention had been paid to the chromosomal distribution and cytogenetic diversity of these sequences. This paper report the long-range organization of all possible classes of mono-, di- and tri-nucleotide SSRs in Brassica rapa. Fluorescence in situ hybridization (FISH was used to characterize the cytogenetic diversity of SSRs among morphotypes of B. rapa ssp. chinensis. The proportion of different SSR motifs varied among morphtypes of B. rapa, with trinucleotide SSRs more prevalent in the genome of B. rapa ssp. chinensis. The chromosomal characterizations of mono-, di- and tri-nucleotide repeats have been acquired. The data has revealed the non-random and motif-dependent chromosome distribution of SSRs in different morphtypes, and allowed the relative variability characterized by SSRs amount and similar chromosomal distribution in centromeric/peri-centromeric heterochromatin. The differences of SSRs in the abundance and distribution indicated the driving force of SSRs in relationship with the evolution of B. rapa species. The results provided a comprehensive view on the SSR sequence distribution and evolution for comparison among morphtypes B. rapa ssp. chinensis.

  4. Simultaneous genotyping of HPA-17w to -21w by PCR-SSP in Chinese Cantonese.

    Science.gov (United States)

    Zhou, Haojie; Ding, Haoqiang; Chen, Yangkai; Li, Xiaofan; Ye, Xin; Nie, Yongmei

    2015-01-01

    Studies have reported the polymorphism of human platelet antigen (HPA)-17w, -18w, -19w, -20w, and -21w. However, the distribution of these five antigens in Chinese Cantonese is still unknown. In this study, we designed new sequence-specific primers for HPA-19w to -21w and used published primers for HPA-17w and -18w to develop a polymerase chain reaction with the sequence-specific primers (PCR-SSP) method for simultaneously genotyping HPA-17w to -21w. A total of 820 unrelated Cantonese apheresis platelet donors in Guangzhou were involved in this study. Among the five HPAs, complete a/a homozygosity was observed for HPA-17w to -20w with an allele frequency of 1.0000. For HPA-21w, nine individuals (9/820, 1.10%) were found to be HPA-21a/bw heterozygous and the allele frequencies of HPA-21a and HPA-21bw were 0.9945 (1631/1640) and 0.0055 (9/1640), respectively. The reliability of the PCR-SSP method was determined by comparing with the genotyping results by DNA sequencing, and no inconsistencies were observed between the two methods. This study provides a reliable PCR-SSP method for simultaneously genotyping HPA-17w to -21w and could improve HPA-matched platelet transfusion in Chinese Cantonese.

  5. Environmental Restoration and Waste Management Site-Specific Plan (SSP) for fiscal year 1992 (FY92)

    International Nuclear Information System (INIS)

    1991-09-01

    The FY-92 Site-Specific Plan (FY-92 SSP) for environmental restoration and waste management at the Idaho National Engineering Laboratory (INEL) is designed to provide the reader with easy access to the status of environmental restoration and waste management activities at INEL. The first chapter provides background on INIEL's physical environment, site history and mission, and general information about the site and its facilities. In addition, this chapter discusses the inter-relationships between the Site Specific Plan, the Environmental Restoration and Waste Management Five-Year Plan, the environmental restoration and waste management prioritization systems, and the Activity Data Sheets (ADSs) for environmental restoration and waste management. This discussion should help readers understand what the SSP is and how it fits into the environmental restoration and waste management process at INEL. This understanding should provide the reader with a better context for understanding the discussions in the SSP as well as a better feel for how and what to comment on during the public comment period that will be held from the first of September through the end of October 1991

  6. Propionic acid fermentation of glycerol and glucose by Propionibacterium acidipropionici and Propionibacterium freudenreichii ssp. shermanii.

    Science.gov (United States)

    Himmi, E H; Bories, A; Boussaid, A; Hassani, L

    2000-04-01

    A comparative study was carried out in anaerobic batch cultures on 20 g/l of either glycerol or glucose using two propionibacteria strains, Propionibacterium acidipropionici and Propionibacterium freudenreichii ssp. shermanii. In all cases, fermentation end-products were the same and consisted of propionic acid as the major product, acetic acid as the main by-product and two minor metabolites, n-propanol and succinic acid. Evidence was provided that greater production of propionic acid by propionibacteria was obtained with glycerol as carbon and energy sources. P. acidipropionici showed higher efficiency in glycerol conversion to propionic acid with a faster substrate consumption (0.64 g l(-1) h(-1)) and a higher propionic acid production (0.42 g l(-1) h(-1) and 0.79 mol/mol). The almost exclusive production of propionic acid from glycerol by this bacterium suggested an homopropionic tendency of this fermentation. Acetic acid final concentration was two times lower on glycerol (2 g/l) than on glucose (4 g/l) for both micro-organisms. P. freudenreichii ssp. shermanii exhibited a glycerol fermentation pattern typical of non-associated glycerol-consumption-product formation. This could indicate a particular metabolism for P. freudenreichii ssp. shermanii oriented towards the production of other specific components. These results tend to show that glycerol could be an excellent alternative to conventional carbon sources such as carbohydrates for propionic acid production.

  7. A nano particle vector comprised of poly lactic-co-glycolic acid and monophosphoryl lipid A and recombinant Mycobacterium avium subsp paratuberculosis peptides stimulate a pro-immune profile in bovine macrophages

    Science.gov (United States)

    Current research and development of antigens for vaccination often center on purified recombinant proteins, viral vectored subunits, and synthetic peptides, most of which suffer from poor immunogenicity and are subject to degradation. For these reasons, efficient delivery systems and potent immunost...

  8. Addendum to “Novel antigens for detection of cell mediated immune responses to Mycobacterium avium subsp. paratuberculosis infection in cattle” [Vet. Immunol. Immunopathol. 143 (2011) 46–54

    DEFF Research Database (Denmark)

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

    2012-01-01

    In the original published paper, the identities of the 14 novel antigens used were not revealed. Their identities are now available to supplement Table 1 in the original paper, as listed in Table S1, which also includes the names of the corresponding M. tuberculosis and M. bovis orthologs. In Table...

  9. Comparison of the results of climate change impact assessment between RCP8.5 and SSP2 scenarios

    Science.gov (United States)

    Lee, D. K.; Park, J. H.; Park, C.; Kim, S.

    2017-12-01

    Climate change scenarios are mainly published by the Intergovernmental Panel on Climate Change (IPCC), and include SRES (Special Report on Emission Scenario) scenarios (IPCC Third Report), RCP (Representative Concentration Pathways) scenarios (IPCC 5th Report), and SSP (Shared Socioeconomic Pathways) scenarios. Currently widely used RCP scenarios are based on how future greenhouse gas concentrations will change. In contrast, SSP scenarios are that predict how climate change will change in response to socio-economic indicators such as population, economy, land use, and energy change. In this study, based on RCP 8.5 climate data, we developed a new Korean scenario using the future social and economic scenarios of SSP2. In the development of the scenario, not only Korea's emissions but also China and Japan's emissions were considered in terms of space. In addition, GHG emissions and air pollutant emissions were taken into consideration. Using the newly developed scenarios, the impacts assessments of the forest were evaluated and the impacts were evaluated using the RCP scenarios. The average precipitation is similar to the SSP2 scenario and the RCP8.5 scenario, but the SSP2 scenario shows the maximum value is lower than RCP8.5 scenario. This is because the SSP2 scenario simulates the summer precipitation weakly. The temperature distribution is similar for both scenarios, and it can be seen that the average temperature in the 2090s is higher than that in the 2050s. At present, forest net primary productivity of Korea is 693 tC/km2, and it is 679 tC/km2 when SSP2 scenario is applied. Also, the damage of forest by ozone is about 4.1-5.1%. On the other hand, when SSP2 scenario is applied, the forest net primary productivity of Korea is 607 tC/km2 and the forest net primary productivity of RCP8.5 scenario is 657 tC/km2. The analysis shows that the damage caused by climate change is reduced by 14.2% for the SSP2 scenario and 6.9% for the RCP8.5 scenario. The damage caused

  10. Utility of gallium imaging in the diagnosis of Mycobacterium avium-intracellulare infection in patients with the acquired immunodeficiency syndrome

    International Nuclear Information System (INIS)

    Bach, M.C.; Bagwell, S.P.; Masur, H.

    1986-01-01

    Whole body Ga-67 scans revealed increased uptake in lymph nodes accessible for biopsy in three patients with the acquired immunodeficiency syndrome (AIDS) infected by Mycobacterium avium-intracellulare (MAI). In diagnostically difficult cases where the usual methods for diagnosing MAI are not helpful, Ga-67 studies may be of value

  11. COMPARISON OF MYCOBACTERIUM AVIUM ISOLATES FROM A DRINKING WATER DISTRIBUTION SYSTEM AND FROM THE POPULATION SERVED BY THE SYSTEM

    Science.gov (United States)

    Background: Current evidence suggests that drinking water, soil, and produce are potential sources of Mycobacterium avium infections, a pathogen not known to be transmitted person-to-person. Methods: We sampled water during 2000-2002 from a large municipal drinking water ...

  12. MOLECULAR COMPARISON OF MYCOBACTERIUM AVIUM ISOLATED FROM A FRINKING WATER DISTRIBUTION SYSTEM AND FROM THE POPULATION SERVED BY THE SYSTEM

    Science.gov (United States)

    There is evidence that drinking water, soil, and produce may be sources of Mycobacterium avium infections, a pathogen not known to be transmitted person-to-person. We sampled water from a large municipal drinking water distribution system in which surface source water is used. M...

  13. Seed washing, exogenous application of gibberellic acid, and cold stratification enhance the germination of sweet cherry (Prunus avium L.) seed

    NARCIS (Netherlands)

    Javanmard, T.; Zamani, Z.; Keshavarz Afshar, R.; Hashemi, M.; Struik, P.C.

    2014-01-01

    Seed germination in sweet cherry (Prunus avium L.) is a slow and lengthy process which has delayed breeding efforts. In this study, seed from ripe fruit of the sweet cherry cultivar ‘Lambert’ were collected and, after removing the endocarp, various dormancy-breaking treatments such as seed washing,

  14. Coexistent Pseudogout and Mycobacterium avium-intracellulare Septic Arthritis in a Patient with HIV and ESRD

    Directory of Open Access Journals (Sweden)

    Wais Afzal

    2016-01-01

    Full Text Available Pseudogout is a crystal-induced arthropathy characterized by the deposition of calcium pyrophosphate dihydrate (CPPD crystals in synovial fluid, menisci, or articular cartilage. Although not very common, this entity can be seen in patients with chronic kidney disease (CKD. Septic arthritis due to Mycobacterium avium-intracellulare (MAI is a rare entity that can affect immunocompromised patients such as those with acquired immunodeficiency syndrome (AIDS or those who are on immunosuppressive drugs. Here, we describe a 51-year-old female who presented with fever, right knee pain, swelling, warmth, and decreased range of motion for several days. The initial assessment was consistent with pseudogout, with negative bacterial and fungal cultures. However, due to high white blood cell (WBC count in the synovial fluid analysis, she was empirically started on intravenous (IV vancomycin and piperacillin-tazobactam and discharged on IV vancomycin and cefepime, while acid-fast bacilli (AFB culture was still in process. Seventeen days later, AFB culture grew Mycobacterium avium-intracellulare (MAI, and she was readmitted for relevant management. This case illustrates that septic arthritis due to MAI should be considered in the differential diagnosis of septic arthritis in immunocompromised patients.

  15. [Mycobacterium avium tumoral infection mimicking a lung adenocarcinoma: A potential diagnostic pitfall].

    Science.gov (United States)

    Fiogbe, A A; Liistro, G; Hoton, D; Pieters, T

    2016-04-01

    The incidence of atypical mycobacterial infection in Europe is estimated at one case per 100,000 persons/year. Despite the low incidence of Mycobacterium avium infection, it can result in a nodular lesion simulating lung cancer. We report a case of atypical mycobacteriosis, mimicking lung cancer, which led to a lobectomy. It was a right pulmonary upper lobe nodule found in a 63-year-old COPD patient, partially nephrectomized for renal carcinoma, and weekly treated by methotrexate for rheumatoid arthritis. FDG uptake was weakly positive on PET-CT (SUV=2.2) in the upper fissure. Bronchoscopy yielded no lesions and no bacteriological findings. Percutaneous transthoracic lung biopsy revealed lung adenocarcinoma stage T1 (a) N0M0. An upper lobectomy with lymphadenectomy was performed. Histological examination revealed epithelioid granuloma surrounded by giant cells suggestive of tuberculomas. The bronchial washing fluid culture was positive for Mycobacterium avium after 7 weeks. In pseudo-neoplastic forms of atypical mycobacteriosis, the presence of alveolar, inflammatory cytonuclear abnormalities can mimic an adenocarcinoma. Making the difference between the cytonuclears defects related to inflammation or neoplasia remains a daily challenge in histopathology. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  16. Evaluation of test-strategies for estimating probability of low prevalence of paratuberculosis in Danish dairy herds

    DEFF Research Database (Denmark)

    Sergeant, E.S.G.; Nielsen, Søren S.; Toft, Nils

    2008-01-01

    -sensitivity and specificity; use of a distribution of observed values (rather than a fixed, low value) for design prevalence; and estimates of the probability of low prevalence (Pr-Low) based on a specific number of test-positive animals, rather than for a result less than or equal to a specified cut-point number of reactors...... of this study was to develop a method to estimate the probability of low within-herd prevalence of paratuberculosis for Danish dairy herds. A stochastic simulation model was developed using the R(R) programming environment. Features of this model included: use of age-specific estimates of test....... Using this model, five herd-testing strategies were evaluated: (1) milk-ELISA on all lactating cows; (2) milk-ELISA on lactating cows 4 years old; (4) faecal culture on all lactating cows; and (5) milk-ELISA plus faecal culture in series on all lactating cows. The five testing strategies were evaluated...

  17. 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 a MAP protein expressed in latent infection (FET11 vaccine). FET11 vaccine proteins were formulated with CAF01 adjuvant and injected to MAP challenged calves at two different ages. 28 calves divided into two FET11 vaccine groups, a commercial vaccine and a control group were used in the study...... and 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...

  18. Paratuberculose em caprinos e ovinos no Brasil Paratuberculosis in goats and sheep in Brazil

    Directory of Open Access Journals (Sweden)

    Diego M. Oliveira

    2010-01-01

    caprinos e ovinos no Brasil, determinar medidas de controle e avaliar a possibilidade de transmissão para humanos.This paper reports paratuberculosis in sheep and goats for the first time in Brazil. On Farm 1 in a flock of 33 goats and 13 sheep, one adult female goat presented progressive weight loss during about one year, and soft feces during one month before death. All the animals from the flock were submitted to the comparative tuberculin test. One sheep was positive to the avian tuberculin (2.2% and in two the results were inconclusive. On Farm 2, in a flock of 200 sheep and 80 goats, one adult ewe presented progressive weight loss during about one year, and soft feces during 20 days before death. All sheep older than four months of age and 23 goats were submitted to tuberculin test with avian tuberculin; 47 (25.4% sheep were positive, 115 (61.5% had inconclusive results, and 25 (13.4% were negative. No positive tuberculin test were observed in the goats, but in 11 (47.8% the test was inconclusive and in 12 (52.2% was negative. At necropsy both affected animals had enlarged mesenteric lymph nodes. In the affected sheep, the gut wall was thickened and corrugated, mainly in the ileum and ileocecal valve. Histologically, granulomas with mainly foamy macrophages were observed in the lamina propria and submucosa of the gut, in mesenteric lymph nodes, and in the goat also in the liver. One tuberculin positive sheep and one tuberculin negative goat from Farm 1 were euthanized and necropsied. A mild thickened of the intestinal wall was observed grossly in both animals. Inflammatory infiltrate with mainly lymphocytes was observed on the submucosa and lamina propria of the gut. In the four cases acid-alcohol positive rods, positive for Mycobacterium spp. on immunohistochemical examination were observed in macrophages and lymphocytes. New studies are necessary to know the prevalence and economical impact of paratuberculosis in goats and sheep in Brazil, to determine control

  19. 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.

  20. Paratuberculosis Vaccination Causes Only Limited Cross-Reactivity in the Skin Test for Diagnosis of Bovine Tuberculosis

    Science.gov (United States)

    Garrido, Joseba M.; Vazquez, Patricia; Molina, Elena; Plazaola, Jose M.; Sevilla, Iker A.; Geijo, Maria V.; Alonso-Hearn, Marta; Juste, Ramon A.

    2013-01-01

    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. PMID:24303029

  1. Detection of Mycobacterium tuberculosis and Mycobacterium avium Complexes by Real-Time PCR in Bovine Milk from Brazilian Dairy Farms.

    Science.gov (United States)

    Bezerra, André Vinícius Andrade; Dos Reis, Emily Marques; Rodrigues, Rogério Oliveira; Cenci, Alexander; Cerva, Cristine; Mayer, Fabiana Quoos

    2015-05-01

    Foodborne diseases are a public health problem worldwide. The consumption of contaminated raw milk has been recognized as a major cause of transmission of bovine tuberculosis to humans. Other mycobacteria that may be present in raw milk and may cause diseases are those belonging to the Mycobacterium avium complex. In this study, molecular biology tools were applied to investigate raw milk contamination with Mycobacterium spp. in family dairy farms from Rio Grande do Sul, southern Brazil. Furthermore, different variables related to the source of the milk, herd characteristics, and management were evaluated for their effect on milk contamination. Five hundred and two samples were analyzed, of which 354 were from the Northwest region (102 farms with samples from 93 bulk tanks and 261 animals) and 148 from the South region of the state (22 farms with samples from 23 bulk tanks and 125 animals). Among them, 10 (1.99%) and 7 (1.39%) were positive for Mycobacterium tuberculosis (9 confirmed as Mycobacterium bovis) and M. avium complexes, respectively. There was no difference in the frequencies of positive samples between the regions or the sample sources. Of the positive samples, 4 were collected from a bulk tank (1 positive for M. avium and 3 for M. tuberculosis). Moreover, 1 sample was positive concomitantly for M. tuberculosis and M. avium complexes. On risk analysis, no variable was associated with raw milk contamination by M. tuberculosis complex species. However, washing the udders of all animals and drying them with paper towels were weakly classified as risk factors for M. avium contamination. Positive samples were obtained from both animals and bulk tanks, which emphasizes the importance of tuberculosis control programs and provides evidence that milk monitoring can be used as a control practice. Moreover, the findings of this study reinforce the need for awareness of the problems of raw milk consumption among the general population.

  2. Surveillance of Bulk Raw and Commercially Pasteurized Cows' Milk from Approved Irish Liquid-Milk Pasteurization Plants To Determine the Incidence of Mycobacterium paratuberculosis

    OpenAIRE

    O'Reilly, Ciara E.; O'Connor, Lisa; Anderson, Wayne; Harvey, Peter; Grant, Irene R.; Donaghy, John; Rowe, Michael; O'Mahony, Pat

    2004-01-01

    Over the 13-month period from October 2000 to November 2001 (inclusive), the Food Safety Authority of Ireland (FSAI) carried out surveillance of Irish bulk raw (n = 389) and commercially pasteurized (n = 357) liquid-milk supplies to determine the incidence of Mycobacterium paratuberculosis. The pasteurization time-temperature conditions were recorded for all pasteurized samples. Overall, 56% of whole-milk pasteurized samples had been heat treated at or above a time-temperature combination of ...

  3. Ordeñando micobacterias del ganado? Impacto económico y en salud de Tuberculosis bovina y Paratuberculosis en Colombia

    Directory of Open Access Journals (Sweden)

    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.

  4. In vitro susceptibility of Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis, Mycobacterium avium, Mycobacterium fortuitum, and Mycobacterium chelonae to ticarcillin in combination with clavulanic acid.

    OpenAIRE

    Casal, M J; Rodriguez, F C; Luna, M D; Benavente, M C

    1987-01-01

    The in vitro susceptibility of Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium africanum, Mycobacterium avium, Mycobacterium fortuitum, and Mycobacterium chelonae (M. chelonei) to ticarcillin in combination with calvulanic acid (CA) was studied by the agar dilution method. All the M. tuberculosis, M. bovis, and M. africanum strains were inhibited at a ticarcillin concentration of 32 micrograms/ml or lower in combination with 5 micrograms of CA. M. chelonae and M. avium strains ...

  5. Comparison of a Variable-Number Tandem-Repeat (VNTR) Method for Typing Mycobacterium avium with Mycobacterial Interspersed Repetitive-Unit-VNTR and IS1245 Restriction Fragment Length Polymorphism Typing▿ †

    OpenAIRE

    Inagaki, Takayuki; Nishimori, Kei; Yagi, Tetsuya; Ichikawa, Kazuya; Moriyama, Makoto; Nakagawa, Taku; Shibayama, Takami; Uchiya, Kei-ichi; Nikai, Toshiaki; Ogawa, Kenji

    2009-01-01

    Mycobacterium avium complex (MAC) infections are increasing annually in various countries, including Japan, but the route of transmission and pathophysiology of the infection remain unclear. Currently, a variable-number tandem-repeat (VNTR) typing method using the Mycobacterium avium tandem repeat (MATR) loci (MATR-VNTR) is employed in Japan for epidemiological studies using clinical isolates of M. avium. In this study, the usefulness of this MATR-VNTR typing method was compared with that of ...

  6. A putative serine protease, SpSsp1, from Saprolegnia parasitica is recognised by sera of rainbow trout, Oncorhynchus mykiss

    Science.gov (United States)

    Minor, Kirsty L.; Anderson, Victoria L.; Davis, Katie S.; Van Den Berg, Albert H.; Christie, James S.; Löbach, Lars; Faruk, Ali Reza; Wawra, Stephan; Secombes, Chris J.; Van West, Pieter

    2014-01-01

    Saprolegniosis, the disease caused by Saprolegnia sp., results in considerable economic losses in aquaculture. Current control methods are inadequate, as they are either largely ineffective or present environmental and fish health concerns. Vaccination of fish presents an attractive alternative to these control methods. Therefore we set out to identify suitable antigens that could help generate a fish vaccine against Saprolegnia parasitica. Unexpectedly, antibodies against S. parasitica were found in serum from healthy rainbow trout, Oncorhynchus mykiss. The antibodies detected a single band in secreted proteins that were run on a one-dimensional SDS-polyacrylamide gel, which corresponded to two protein spots on a two-dimensional gel. The proteins were analysed by liquid chromatography tandem mass spectrometry. Mascot and bioinformatic analysis resulted in the identification of a single secreted protein, SpSsp1, of 481 amino acid residues, containing a subtilisin domain. Expression analysis demonstrated that SpSsp1 is highly expressed in all tested mycelial stages of S. parasitica. Investigation of other non-infected trout from several fish farms in the United Kingdom showed similar activity in their sera towards SpSsp1. Several fish that had no visible saprolegniosis showed an antibody response towards SpSsp1 suggesting that SpSsp1 might be a useful candidate for future vaccination trial experiments. PMID:25088077

  7. Identification of Two Novel Mycobacterium avium Allelic Variants in Pig and Human Isolates from Brazil by PCR-Restriction Enzyme Analysis

    Science.gov (United States)

    Leão, Sylvia Cardoso; Briones, Marcelo R. S.; Sircili, Marcelo Palma; Balian, Simone Carvalho; Mores, Nelson; Ferreira-Neto, José Soares

    1999-01-01

    Mycobacterium avium complex (MAC) is composed of environmental mycobacteria found widely in soil, water, and aerosols that can cause disease in animals and humans, especially disseminated infections in AIDS patients. MAC consists of two closely related species, M. avium and M. intracellulare, and may also include other, less-defined groups. The precise differentiation of MAC species is a fundamental step in epidemiological studies and for the evaluation of possible reservoirs for MAC infection in humans and animals. In this study, which included 111 pig and 26 clinical MAC isolates, two novel allelic M. avium PCR-restriction enzyme analysis (PRA) variants were identified, differing from the M. avium PRA prototype in the HaeIII digestion pattern. Mutations in HaeIII sites were confirmed by DNA sequencing. Identification of these isolates as M. avium was confirmed by PCR with DT1-DT6 and IS1245 primers, nucleic acid hybridization with the AccuProbe system, 16S ribosomal DNA sequencing, and biochemical tests. The characterization of M. avium PRA variants can be useful in the elucidation of factors involved in mycobacterial virulence and routes of infection and also has diagnostic significance, since they can be misidentified as M. simiae II and M. kansasii I if the PRA method is used in the clinical laboratory for identification of mycobacteria. PMID:10405407

  8. RNA-seq Analysis of Cold and Drought Responsive Transcriptomes of Zea mays ssp. mexicana L.

    Science.gov (United States)

    Lu, Xiang; Zhou, Xuan; Cao, Yu; Zhou, Meixue; McNeil, David; Liang, Shan; Yang, Chengwei

    2017-01-01

    The annual Zea mays ssp. mexicana L. is a member of teosinte, a wild relative of the Zea mays spp. mays L. This subspecies has strong growth and regeneration ability, high tiller numbers, high protein and lysine content as well as resistance to many fungal diseases, and it can be effectively used in maize improvement. In this study, we reported a Zea mays ssp. mexicana L. transcriptome by merging data from untreated control (CK), cold (4°C) and drought (PEG2000, 20%) treated plant samples. A total of 251,145 transcripts (N50 = 1,269 bp) and 184,280 unigenes (N50 = 923 bp) were predicted, which code for homologs of near 47% of the published maize proteome. Under cold conditions, 2,232 and 817 genes were up-regulated and down-regulated, respectively, while fewer genes were up-regulated (532) and down-regulated (82) under drought stress, indicating that Zea mays ssp. mexicana L. is more sensitive to the applied cold rather than to the applied drought stresses. Functional enrichment analyses identified many common or specific biological processes and gene sets in response to drought and cold stresses. The ABA dependent pathway, trehalose synthetic pathway and the ICE1-CBF pathway were up-regulated by both stresses. GA associated genes have been shown to differentially regulate the responses to cold in close subspecies in Zea mays. These findings and the identified functional genes can provide useful clues for improving abiotic stress tolerance of maize. PMID:28223998

  9. Global cost analysis on adaptation to sea level rise based on RCP/SSP scenarios

    Science.gov (United States)

    Kumano, N.; Tamura, M.; Yotsukuri, M.; Kuwahara, Y.; Yokoki, H.

    2017-12-01

    Low-lying areas are the most vulnerable to sea level rise (SLR) due to climate change in the future. In order to adapt to SLR, it is necessary to decide whether to retreat from vulnerable areas or to install dykes to protect them from inundation. Therefore, cost- analysis of adaptation using coastal dykes is one of the most essential issues in the context of climate change and its countermeasures. However, few studies have globally evaluated the future costs of adaptation in coastal areas. This study tries to globally analyze the cost of adaptation in coastal areas. First, global distributions of projected inundation impacts induced by SLR including astronomical high tide were assessed. Economic damage was estimated on the basis of the econometric relationship between past hydrological disasters, affected population, and per capita GDP using CRED's EM-DAT database. Second, the cost of adaptation was also determined using the cost database and future scenarios. The authors have built a cost database for installed coastal dykes worldwide and applied it to estimating the future cost of adaptation. The unit costs of dyke construction will increase with socio-economic scenario (SSP) such as per capita GDP. Length of vulnerable coastline is calculated by identifying inundation areas using ETOPO1. Future cost was obtained by multiplying the length of vulnerable coastline and the unit cost of dyke construction. Third, the effectiveness of dyke construction was estimated by comparing cases with and without adaptation.As a result, it was found that incremental adaptation cost is lower than economic damage in the cases of SSP1 and SSP3 under RCP scenario, while the cost of adaptation depends on the durability of the coastal dykes.

  10. Full Scale Test SSP 34m blade, Combined load. Data report

    DEFF Research Database (Denmark)

    Nielsen, Per Hørlyk; Nielsen, Magda; Jensen, Find Mølholt

    This report is part of the research project entitled “Eksperimentel vingeforskning: Strukturelle mekanismer i nutidens og fremtidens store vinger under kombineret last” where a 34m wind turbine blade from SSP-Technology A/S was tested in combined flap and edgewise load. The applied load is 55......% of an imaginary extreme event based on the certification load of the blade. This report describes the reason for choosing the loads and the load direction and the method of applying the loads to the blade. A novel load introduction allows the blade to deform in a more realistic manner, allowing the observation...

  11. Chemical Composition and Possible in Vitro Phytotoxic Activity of Helichrsyum italicum (Roth Don ssp. italicum

    Directory of Open Access Journals (Sweden)

    Vincenzo De Feo

    2011-09-01

    Full Text Available The chemical composition of the essential oil of Helichrysum italicum (Roth Don ssp. italicum, collected in the National Park of Cilento and Diano Valley, Southern Italy, was studied by means of GC and GC/MS. Forty four compounds of 45 constituents were identified in the oil, mainly oxygenated sesquiterpenes. The essential oil was evaluated for its potential in vitro phytotoxic activity against germination and early radicle elongation of radish and garden cress. The radicle elongation of radish was significantly inhibited at the highest doses tested, while germination of both seeds was not affected.

  12. FINE ROOT DISTRIBUTION OF Eucalyptus globulus ssp. maidenii IN RELATION TO SOME SOIL PROPERTIES

    Directory of Open Access Journals (Sweden)

    Juan José Gaitán

    2010-08-01

    Full Text Available A study was conducted to determine the vertical distribution of Eucalyptus globulus ssp. maidenii fine root (with diameters of up to 2 mm density to a depth of  0,5 m and their relationships with soil properties. Roots were sampled near the stem of 10 trees with the soil corer method. Fine root distribution is influenced by soil physical and chemical properties. A very high density of fine roots was found at 20 cm depht,  while a low density was observed in the clayey horizon. Fine root density (DRF correlated positively with organic C and extractable P and negatively with soil bulk density.

  13. Lineages of Streptococcus equi ssp. equi in the Irish equine industry.

    Science.gov (United States)

    Moloney, Emma; Kavanagh, Kerrie S; Buckley, Tom C; Cooney, Jakki C

    2013-01-01

    Streptococcus equi ssp. equi is the causative agent of 'Strangles' in horses. This is a debilitating condition leading to economic loss, yard closures and cancellation of equestrian events. There are multiple genotypes of S. equi ssp. equi which can cause disease, but to date there has been no systematic study of strains which are prevalent in Ireland. This study identified and classified Streptococcus equi ssp. equi strains isolated from within the Irish equine industry. Two hundred veterinary isolates were subjected to SLST (single locus sequence typing) based on an internal sequence from the seM gene of Streptococcus equi ssp equi. Of the 171 samples which successfully gave an amplicon, 162 samples (137 Irish and 24 UK strains) gave robust DNA sequence information. Analysis of the sequences allowed division of the isolates into 19 groups, 13 of which contain at least 2 isolates and 6 groups containing single isolates. There were 19 positions where a DNA SNP (single nucleotide polymorphism) occurs, and one 3 bp insertion. All groups had multiple (2-8) SNPs. Of the SNPs 17 would result in an amino acid change in the encoded protein. Interestingly, the single isolate EI8, which has 6 SNPs, has the three base pair insertion which is not seen in any other isolate, this would result in the insertion of an Ile residue at position 62 in that protein sequence. Comparison of the relevant region in the determined sequences with the UK Streptococcus equi seM MLST database showed that Group B (15 isolates) and Group I (2 isolates), as well as the individual isolates EI3 and EI8, are unique to Ireland, and some groups are most likely of UK origin (Groups F and M), but many more probably passed back and forth between the two countries. The strains occurring in Ireland are not clonal and there is a considerable degree of sequence variation seen in the seM gene. There are two major clades causing infection in Ireland and these strains are also common in the UK.

  14. Antispasmodic effects and structure-activity relationships of labdane diterpenoids from Marrubium globosum ssp. libanoticum.

    Science.gov (United States)

    Rigano, Daniela; Aviello, Gabriella; Bruno, Maurizio; Formisano, Carmen; Rosselli, Sergio; Capasso, Raffaele; Senatore, Felice; Izzo, Angelo A; Borrelli, Francesca

    2009-08-01

    Marrubium globosum ssp. libanoticum is a medicinal plant used in Lebanon to reduce pain and smooth muscle spasms. A chloroform extract obtained from M. globosum aerial parts reduced acetylcholine-induced contractions in the isolated mouse ileum. The purification of this extract identified, among 12 isolated labdane diterpenoids, four new compounds, named 13-epicyllenin A (4), 13,15-diepicyllenin A (5), marrulibacetal (9), and marrulactone (11). Their structures were determined by spectroscopic methods. Compound 9, which exerted antispasmodic activity, is likely the active ingredient of the extract. Preliminary structure-activity relationships for this class of compounds are suggested.

  15. Phytochemical and pharmacological studies on the acetonic extract of Marrubium globosum ssp. libanoticum.

    Science.gov (United States)

    Rigano, Daniela; Grassia, Armando; Borrelli, Francesca; Aviello, Gabriella; Piozzi, Franco; Bruno, Maurizio; Arnold, Nelly Apostolides; Capasso, Raffaele; Senatore, Felice

    2006-05-01

    A new natural labdane diterpene, marrulibanoside, was isolated from the acetonic extract of aerial parts of M. globosum Montbr. et Auch. ex Benth. ssp. libanoticum Boiss. (Lamiaceae). Its structure was determined by spectroscopic methods such as 1D and 2D NMR and mass spectrometry. Pharmacological studies have shown that the extract of M. globosum exerts anti-inflammatory effects in the rat paw oedema induced by carrageenin resulting in reduced paw swelling. This activity, which seems due to marrulibanoside, is a consequence of iNOS and COX-2 activities inhibition.

  16. PENGEMBANGAN SUBJECT SPECIFIC PEDAGOGY (SSP IPA TERPADU UNTUK MENINGKATKAN HASIL BELAJAR SISWA

    Directory of Open Access Journals (Sweden)

    Fitri Yuliawati

    2016-05-01

    Full Text Available Background of this research is the analysis of the results of interviews some junior secondary schools in Yogyakarta and the conclusion that teachers do not use an integrated science teaching and teachers still find difficulties in the application of learning science in an integrated manner. It can be influenced by several factors, such as the lack of reference used by teachers in presenting the material Integrated Science relevantly, most of science teachers are from educational background of chemistry, physics, and biology instead of science education, so that teachers find difficulties to create an integrated learning of science. In addition, teachers feel difficulty in determining the depth of the material, limits of integration in integrated science teaching, and did not know the concept of integrated science teaching. This research is a development research. Learning tools developed included : student books, lesson plans, student activity sheets and evaluation tools. The development of the learning which is done in this study use the models of 4D development which includes the step of Define (definition which at this stage conducted a needs analysis. Design : it is the stage of Subject Specific Pedagogy (SSP software design. Development, it is the stage of development after the draft was made followed by a learning device validation by experts. This stage is also conducted to seek input from all the responses, reactions and comments from teachers, students, and observers so that it can be used for further improvement of science teaching later. The Disseminate, it is the stage of field tests are widely but not done. Data collection instruments used in this study include : test items and questionnaire. The conclusion of this development research are as follows : the results of the validation SSP integrated science by learning tools expert, material experts and media experts indicate the category of Very Good (SB so that SSP integrated

  17. EVALUACIÓN DE UNA COLECCIÓN DEL GÉNERO Triticum: TRIGO HARINERO (Triticum aestivum ssp aestivum), TRIGO DURO (Triticum turgidum ssp. durum) Y TRITICALE (X Triticum secale Wittmack) EN LAS CONDICIONES DEL OCCIDENTE DE CUBA

    OpenAIRE

    R. Plana; Marta Álvarez; M. Varela

    2006-01-01

    En el Instituto Nacional de Ciencias Agrícolas (INCA) se evaluó una colección del género Triticum, con el objetivo de conocer el comportamiento de diferentes especies en las condiciones del occidente de Cuba. Las especies estudiadas fueron: Triticum aestivum ssp. aestivum, Triticum turgidum ssp. durum y X Triticum secale Wittmack. Se utilizó un diseño experimental de bloques al azar con cuatro repeticiones. Los caracteres analizados fueron altura de la planta (cm), longitud de las espigas (cm...

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

    LENUS (Irish Health Repository)

    Judge, EP

    2016-04-01

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

  19. Mycobacterium avium intracellulare complex causing olecranon bursitis and prosthetic joint infection in an immunocompromised host

    Directory of Open Access Journals (Sweden)

    Eugene M. Tan

    2016-01-01

    Full Text Available Case: A 73-year-old immunocompromised male presented with recurrent left elbow swelling due to Mycobacterium avium intracellulare complex (MAC olecranon bursitis. 3 years after completing MAC treatment, he underwent right total knee arthroplasty (TKA. 1 year later, he developed TKA pain and swelling and was diagnosed with MAC prosthetic joint infection (PJI. He underwent TKA resection, reimplantation, and 12 months of anti-MAC therapy. This patient is the seventh case report of MAC olecranon bursitis and the third case report of MAC PJI. He is the only report of both MAC olecranon bursitis and PJI occurring in the same patient. Informed consent: This patient was informed and agreed to the publication of this material.

  20. Opportunistic Pathogens Mycobacterium Avium Complex (MAC) and Legionella spp. Colonise Model Shower.

    Science.gov (United States)

    Whiley, Harriet; Giglio, Steven; Bentham, Richard

    2015-07-24

    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.

  1. Phenolic compounds, antioxidant and antimicrobial properties of the wild cherry (Prunus avium L. stem

    Directory of Open Access Journals (Sweden)

    Ademović Zahida

    2017-01-01

    Full Text Available The aim of this study was to determine the total phenolic content, evaluate antioxidant propertie and antimicrobial potential, and identify phenolic compounds in alcoholic and aqueous extracts of the wild cherry (Prunus avium L. stems collected in Bosnia and Herzegovina. Alcoholic extracts had higher contents of phenolic and flavonoid components, as well as the antioxidant and ferric reducing antioxidant capacity in comparison to aqueous extracts. All extracts were characterized by HPLC analysis. Furthermore, for the first time, the antimicrobial properties of wild cherry stem extracts were evaluated. Quercetin and (+-catechin were the main compounds identified in the alcoholic extract, followed by chlorogenic acid and rutin. Quercetin was also the major component detected in aqueous extracts. Besides, alcoholic extract showed better antibacterial properties against Staphylococcus aureus as a representative gram-positive bacteria than infusion, whereas none of the samples showed antibacterial activity against Escherichia coli and fungus Candida albicans.

  2. Biological Evaluation, Overpressured Layer Chromatography (OPLC) separation and Isolation of a New Acetylenic Derivative Compound from Prangos platychlaena ssp. platychlaena Fruit Essential Oils

    Science.gov (United States)

    In the present work, fruits of Prangos platychlaena Boiss. ex Tchihat. ssp. platychlaena (Apiaceae) collected from eastern (Sample A) and central (Sample B) parts of Turkey were subjected to hydrodistillation to yield essential oils (EOs). The chemical composition of P. platychlaena ssp. platychlaen...

  3. 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

  4. 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

  5. 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

  6. Removal of Mycobacterium avium subspecies hominissuis (MAH) from drinking water by coagulation, flocculation and sedimentation processes.

    Science.gov (United States)

    Wong, E A; Shin, G-A

    2015-03-01

    There has been a growing concern over human exposure to Mycobacterium avium subspecies hominissuis (MAH) through drinking water due to its ubiquitous presence in natural waters and remarkable resistance to both chemical and physical disinfectants in drinking water treatment processes. However, little is known about the effectiveness of physico-chemical water treatment processes to remove MAH. Therefore, we determined the removal of MAH by alum coagulation, flocculation and sedimentation processes in optimized drinking water treatment conditions using standard jar test equipment. Contrary to the prevailing hypothesis, the results of this study show that removal of MAH by coagulation, flocculation and sedimentation processes was only moderate (approx. 0.65 log10) under low turbidity treatment conditions and the removal of MAH was actually lower than that of Escherichia coli (reference bacterium) in all the waters tested. Overall, the results of this study suggested that coagulation, flocculation and sedimentation processes may not be a reliable treatment option for removing MAH, and more efforts to find an effective control measures against MAH should be made to reduce the risk of MAH infection from drinking water. Despite a growing concern over human exposure to Mycobacterium avium subspecies hominissuis (MAH) through drinking water and its remarkable resistance to water disinfectants, little is known about the effectiveness of physico-chemical water treatment processes to remove MAH. Contrary to the prevailing hypothesis, the results of this study suggest that coagulation, flocculation and sedimentation processes may not be a reliable treatment option for MAH removal. As these processes have been the last remaining conventional drinking water treatment processes that might be effective against MAH, more efforts should be urgently made to find an effective control measures against this important waterborne pathogen. © 2014 The Society for Applied Microbiology.

  7. Development of a typing system for epidemiological studies of porcine toxin-producing Pasteurella multocida ssp. multocida in Denmark

    DEFF Research Database (Denmark)

    Fussing, V.; Nielsen, Jens; Bisgaard, M.

    1999-01-01

    The aim of the present study was to evaluate capsular-typing, plasmid-profiling, phage-typing and ribotyping for epidemiological studies of toxin-producing Pasteurella multocida ssp. multocida in Denmark. The evaluation of methods was based on 68 strains from nasal swabs and 14 strains from...... by HindIII ribotyping, as 85% of isolates from all herds were assigned to one ribotype. In conclusion, HindIII ribotyping seems to represent a useful tool for epidemiological studies of toxigenic P. multocida ssp. multocida....

  8. Congenital vestibular disease in captive Sumatran tigers (Panthera tigris ssp. sumatrae) in Australasia.

    Science.gov (United States)

    Wheelhouse, Jaimee L; Hulst, Frances; Beatty, Julia A; Hogg, Carolyn J; Child, Georgina; Wade, Claire M; Barrs, Vanessa R

    2015-11-01

    The Sumatran tiger (Panthera tigris ssp. sumatrae) is a critically endangered species in the wild. To ensure that demographic and genetic integrity are maintained in the longer term, those Sumatran tigers held in captivity are managed as a global population under a World Association of Zoos and Aquariums Global Species Management Plan (GSMP). A retrospective study, including segregation and pedigree analysis, was conducted to investigate potential cases of congenital vestibular disease (CVD) in captive Sumatran tigers in Australasian zoos using medical and husbandry records, as well as video footage obtained from 50 tigers between 1975 and 2013. Data from the GSMP Sumatran tiger studbook were made available for pedigree and segregation analysis. Fourteen cases of CVD in 13 Sumatran tiger cubs and one hybrid cub (Panthera tigris ssp. sumatrae × Panthera tigris) were identified. Vestibular signs including head tilt, circling, ataxia, strabismus and nystagmus were observed between birth and 2 months of age. These clinical signs persisted for a median of 237 days and had resolved by 2 years of age in all cases. Pedigree analysis revealed that all affected tigers were closely related and shared a single common ancestor in the last four generations. A genetic cause for the disease is suspected and, based on pedigree and segregation analysis, an autosomal dominant mode of inheritance is likely. Further investigations to determine the world-wide prevalence and underlying pathology of this disorder are warranted. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Effect of plant age on PCB accumulation by Cucurbita pepo ssp. pepo.

    Science.gov (United States)

    Low, Jennifer E; Whitfield Aslund, Melissa L; Rutter, Allison; Zeeb, Barbara A

    2010-01-01

    A greenhouse experiment was conducted to investigate polychlorinated biphenyl (PCB) uptake and translocation from soil over time in pumpkin plants (Cucurbita pepo ssp. pepo cv. Howden). Plants were grown in weathered soil collected from a former industrial site contaminated with Aroclor 1248 (mean [PCB](soil) = 6.5 mg kg(-1)). Plants were harvested five times over 42 d and analyzed for total PCB concentration in the root and shoot tissues. The concentration of PCBs in the root was not significantly different between harvests (mean [PCB](root) = 21.5 mg kg(-1)). The concentration of PCBs in the shoots was also relatively stable over time (mean [PCB](shoot) = 3.5 mg kg(-1)) despite increases in shoot biomass (fresh weight of 4.3 g at Day 12 to 59 g at Day 42). This suggests that PCBs were continuously accumulated throughout the growth period. The trends found in this study indicate the optimal time to harvest C. pepo ssp pepo plants to maximize PCB phytoextraction is when the plant shoot has reached its maximum biomass.

  10. Identification of (+)-erythro-mefloquine as an active enantiomer with greater efficacy than mefloquine against Mycobacterium avium infection in mice.

    Science.gov (United States)

    Bermudez, Luiz E; Inderlied, Clark B; Kolonoski, Peter; Chee, Christopher B; Aralar, Priscilla; Petrofsky, Mary; Parman, Toufan; Green, Carol E; Lewin, Anita H; Ellis, William Y; Young, Lowell S

    2012-08-01

    Infection caused by Mycobacterium avium is common in AIDS patients who do not receive treatment with highly active antiretroviral therapy (HAART) or who develop resistance to anti-HIV therapy. Mefloquine, a racemic mixture used for malaria prophylaxis and treatment, is bactericidal against M. avium in mice. MICs of (+)-erythro-, (-)-erythro-, (+)-threo-, and (-)-threo-mefloquine were 32 μg/ml, 32 μg/ml, 64 μg/ml, and 64 μg/ml, respectively. The postantibiotic effect for (+)-erythro-mefloquine was 36 h (MIC) and 41 h for a concentration of 4× MIC. The mefloquine postantibiotic effect was 25 h (MIC and 4× MIC). After baseline infection was established (7 days), the (+)- and (-)-isomers of the diastereomeric threo- and erythro-α-(2-piperidyl)-2,8-bis(trifluoromethyl)-4-quinolinemethanol were individually used to orally treat C57BL/6 bg(+)/bg(+) beige mice that were infected intravenously with M. avium. Mice were also treated with commercial mefloquine and diluent as controls. After 4 weeks of treatment, the mice were harvested, and the number of bacteria in spleen and liver was determined. Mice receiving (+)- or (-)-threo-mefloquine or (-)-erythro-mefloquine had numbers of bacterial load in tissues similar to those of untreated control mice at 4 weeks. Commercial mefloquine had a bactericidal effect. However, mice given the (+)-erythro-enantiomer for 4 weeks had a significantly greater reduction of bacterial load than those given mefloquine. Thus, (+)-erythro-mefloquine is the active enantiomer of mefloquine against M. avium and perhaps other mycobacteria.

  11. The clinical efficacy of a clarithromycin-based regimen for Mycobacterium avium complex disease: A nationwide post-marketing study.

    Science.gov (United States)

    Kadota, Jun-Ichi; Kurashima, Atsuyuki; Suzuki, Katsuhiro

    2017-05-01

    The revised 2007 American Thoracic Society/Infectious Diseases Society of America statement recommend clarithromycin-based combination therapy for treatment of Mycobacterium avium complex lung disease and stipulates approximately 1 year of continuous treatment after bacilli negative conversion. However, supporting data are insufficient. Our objective was to obtain data on the clinical outcome of clarithromycin-based daily regimens by conducting a nationwide retrospective post-marketing study of M. avium complex lung disease. In accordance with the Japanese guidelines, patients were enrolled in this survey according to their chest radiographic findings and microbiologic test results. They were treated with a multidrug regimen including clarithromycin, rifampicin, and ethambutol (clarithromycin-based regimen) until bacilli negative conversion, and the treatment was continued for approximately 1 year after the initial conversion. Data were collected before administration, at the time of bacilli negative conversion, at the end of treatment, and at 6 months after the end of treatment. Of the 466 subjects enrolled in the study, 271 patients who received clarithromycin at 800 mg/day underwent evaluation for M. avium complex disease. The final bacilli negative conversion rate in those patients was 94.7%. The bacteriological relapse rate was 5.0% (5/100 patients). Bacteriological relapse was noted in patients treated for less than 15 months after conversion. No life-threatening or serious adverse drug reactions were observed. This study demonstrated that a clarithromycin-based daily regimen can yield a high bacteriological conversion rate in M. avium complex disease. After conversion, treatment for less than 15 months might be insufficient to prevent bacteriological relapse. Copyright © 2017 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  12. Post-storage cell wall metabolism in two sweet cherry (Prunus avium L.) cultivars displaying different postharvest performance

    OpenAIRE

    Belge, Burcu; Comabella, Eva; Graell i Sarle, Jordi; Lara Ayala, Isabel

    2015-01-01

    The biochemical processes underlying firmness loss of sweet cherry (Prunus avium L.) fruit are poorly understood.Studies on cell wall metabolism of sweet cherry have been generally undertaken during on-tree development or at harvest maturity, while published reports on postharvest changes are scarce and fragmentary. In this work, cell wall modifications after storage at 0 ºC were studied in two cherry cultivars ('Celeste' and 'Somerset') displaying different postharvest potential. Firmness wa...

  13. Detection of Mycobacteria, Mycobacterium avium Subspecies, and Mycobacterium tuberculosis Complex by a Novel Tetraplex Real-Time PCR Assay

    Science.gov (United States)

    Molina, Elena; Elguezabal, Natalia; Pérez, Valentín; Garrido, Joseba M.

    2015-01-01

    Mycobacterium tuberculosis complex, Mycobacterium avium, and many other nontuberculous mycobacteria are worldwide distributed microorganisms of major medical and veterinary importance. Considering the growing epidemiologic significance of wildlife-livestock-human interrelation, developing rapid detection tools of high specificity and sensitivity is vital to assess their presence and accelerate the process of diagnosing mycobacteriosis. Here we describe the development and evaluation of a novel tetraplex real-time PCR for simultaneous detection of Mycobacterium genus, M. avium subspecies, and M. tuberculosis complex in an internally monitored single assay. The method was evaluated using DNA from mycobacterial (n = 38) and nonmycobacterial (n = 28) strains, tissues spiked with different CFU amounts of three mycobacterial species (n = 57), archival clinical samples (n = 233), and strains isolated from various hosts (n = 147). The minimum detectable DNA amount per reaction was 50 fg for M. bovis BCG and M. kansasii and 5 fg for M. avium subsp. hominissuis. When spiked samples were analyzed, the method consistently detected as few as 100 to 1,000 mycobacterial CFU per gram. The sensitivity and specificity values for the panel of clinical samples were 97.5 and 100% using a verified culture-based method as the reference method. The assays performed on clinical isolates confirmed these results. This PCR was able to identify M. avium and M. tuberculosis complex in the same sample in one reaction. In conclusion, the tetraplex real-time PCR we designed represents a highly specific and sensitive tool for the detection and identification of mycobacteria in routine laboratory diagnosis with potential additional uses. PMID:25588660

  14. Molecular characterization of Mycobacterium avium subspecies hominissuis isolated from humans, cattle and pigs in the Uganda cattle corridor using VNTR analysis.

    Science.gov (United States)

    Muwonge, Adrian; Oloya, James; Kankya, Clovice; Nielsen, Sigrun; Godfroid, Jacques; Skjerve, Eystein; Djønne, Berit; Johansen, Tone B

    2014-01-01

    Members of the Mycobacterium avium complex (MAC) cause disease in both human and animals. Their ubiquitous nature makes them both successful microbes and difficult to source track. The precise characterization of MAC species is a fundamental step in epidemiological studies and evaluating of possible reservoirs. This study aimed at identifying and characterizing Mycobacterium avium subsp. hominissuis isolated from human, slaughter cattle and pigs in various parts of the Uganda cattle corridor (UCC) at two temporal points using variable number of tandem repeat (VNTR) analysis. A total of 46 M. avium isolates; 31 from 997 pigs, 12 from 43 humans biopsies and three from 61 cattle lesions were identified to subspecies level using IS1245 and IS901 PCR, thereafter characterized using VNTR. Twelve loci from two previously described VNTR methods were used and molecular results were analyzed and interpreted using Bionumerics 6.1. 37 of the isolates were identified as M. avium subsp. hominissuis and four as M. avium subsp. avium, while five could not be differentiated, possibly due to mixed infection. There was distinct clustering that coincides with the temporal and spatial differences of the isolates. The isolates from humans and cattle in the North Eastern parts of the UCC shared identical VNTR genotypes. The panel of loci gave an overall discriminatory power of 0.88. Some loci were absent in several isolates, probably reflecting differences in isolates from Uganda/Africa compared to isolates previously analyzed by these methods in Europe and Asia. The findings indicate a molecular difference between M. avium subsp. hominissuis isolates from pigs in Mubende and cattle and human in the rest of the UCC. Although human and cattle shared VNTR genotypes in the North Eastern parts of the UCC, it is most likely a reflection of a shared environmental source. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. 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.

  16. IS1245 restriction fragment length polymorphism typing of Mycobacterium avium from patients admitted to a reference hospital in Campinas, Brazil

    Directory of Open Access Journals (Sweden)

    A.C. Panunto

    2003-10-01

    Full Text Available Mycobacterium avium is an important pathogen among immunodeficient patients, especially patients with AIDS. The natural history of this disease is unclear. Several environmental sources have been implicated as the origin of this infection. Polyclonal infection with this species is observed, challenging the understanding of its pathogenesis and treatment. In the present study 45 M. avium strains were recovered from 39 patients admitted to a reference hospital between 1996 and 1998. Species identification was performed using a species-specific nucleic acid hybridization test (AccuProbe® from Gen-Probe®. Strains were genotyped using IS1245 restriction fragment length polymorphism typing. Blood was the main source of the organism. In one patient with disseminated disease, M. avium could be recovered more than once from potentially sterile sites. Strains isolated from this patient had different genotypes, indicating that the infection was polyclonal. Four patient clones were characterized in this population, the largest clone being detected in eight patients. This finding points to a common-source transmission of the organism.

  17. Mycobacterium avium MAV2052 protein induces apoptosis in murine macrophage cells through Toll-like receptor 4.

    Science.gov (United States)

    Lee, Kang-In; Choi, Han-Gyu; Son, Yeo-Jin; Whang, Jake; Kim, Kwangwook; Jeon, Heat Sal; Park, Hye-Soo; Back, Yong Woo; Choi, Seunga; Kim, Seong-Woo; Choi, Chul Hee; Kim, Hwa-Jung

    2016-04-01

    Mycobacterium avium and its sonic extracts induce apoptosis in macrophages. However, little is known about the M. avium components regulating macrophage apoptosis. In this study, using multidimensional fractionation, we identified MAV2052 protein, which induced macrophage apoptosis in M. avium culture filtrates. The recombinant MAV2052 induced macrophage apoptosis in a caspase-dependent manner. The loss of mitochondrial transmembrane potential (ΔΨm), mitochondrial translocation of Bax, and release of cytochrome c from mitochondria were observed in macrophages treated with MAV2052. Further, reactive oxygen species (ROS) production was required for the apoptosis induced by MAV2052. In addition, ROS and mitogen-activated protein kinases were involved in MAV2052-mediated TNF-α and IL-6 production. ROS-mediated activation of apoptosis signal-regulating kinase 1 (ASK1)-JNK pathway was a major signaling pathway for MAV2052-induced apoptosis. Moreover, MAV2052 bound to Toll-like receptor (TLR) 4 molecule and MAV2052-induced ROS production, ΔΨm loss, and apoptosis were all significantly reduced in TLR4(-/-) macrophages. Altogether, our results suggest that MAV2052 induces apoptotic cell death through TLR4 dependent ROS production and JNK pathway in murine macrophages.

  18. Pengembangan Subject Specific Pedagogy (SSP Tematik Berbasis Local Wisdom Untuk Membangun Karakter Hormat dan Kepedulian Siswa SD

    Directory of Open Access Journals (Sweden)

    Amaliyah Ulfah

    2018-01-01

    Full Text Available This study aims at generating possible thematic Subject Specific Pedagogy (SSP based on local wisdom and examining the results teachers and students perception on the pedagogical approach developed in the preliminary field testing. Drawing upon the notion of research and development by Borg & Gall, this research was conducted through different stages involving information gathering, planning, developing preliminary product, preliminary field testing, and revising. The subjects were a small group of second grade students of SD Muhammadiyah Bodon and SD Muhammadiyah Sidoarum Yogyakarta. Each consists of 10 students. The data gathering instrument consists of product validation, character observation sheets, and teacher assessment sheets. The data gathering instrument consists of product validation tool, character observation sheets, and teachers’ assessment sheets. The SSP product generated in this study consists of lesson plans, teaching materials, worksheets, and expert perception. According to the experts of media, materials, and evaluation, the SSP product is in good categories. First grade teachers result a good score to the syllabus and lesson plans, while the worksheets and evaluation are in very good category. The results from the students responses to the textbooks is 78, which is in very good category, while the students worksheets score 71 which includes in excellent category. Therefore, the developed thematic SSP based on local wisdom can be declared appropriate for use in learning.

  19. Introgression of genetic material from Zea mays ssp. Mexicana into cultivated maize was facilitated by tissue culture

    International Nuclear Information System (INIS)

    Wang, L.; Gu, X.; Qu, M.; Luan, J.; Zhang, J.

    2012-01-01

    Zea mays ssp. mexicana, a wild relative of cultivated maize (Z. mays ssp. mays), is a useful gene resource for maize breeding. In this study, two populations were generated by conventional breeding scheme (population I) or tissue culture regime (population II), respectively, to introgress genetic material of Z. mays ssp. mexicana into maize. Karyotype analysis showed that the arm ratios of 10 pairs of chromosomes in parent maize Ye515 and derivative lines from 2 different populations with 26% and 38% chromosome variation frequencies, respectively. Alien chromatin was detected in the root tip cells of progeny plants through genomic in situ hybridization (GISH). There were 3.3 chromosomes carrying alien chromatin on average in population I and 6.5 in population II. The hybridization signals were located mainly at the terminal or sub terminal regions of the chromosomes and the sizes were notably variant among lines. Based on those results, it is concluded that the introgression of genetic material from Z. mays ssp. mexicana into cultivated maize was facilitated by tissue culture, and subsequently some excellent materials for maize breeding were created. (author)

  20. CITOGENETICS EFFECTS INDUCED BY THE ASCORBIC ACID TREATMENT OF LARIX DECIDUA MILL. SSP. CARPATICA AND PICEA ABIES (L. KARST

    Directory of Open Access Journals (Sweden)

    Ioana Ieremia

    2006-08-01

    Full Text Available The paper present the influence of ascorbic acid upon the mitotic division of Larix decidua Mill ssp. carpatica and Picea abies (L. Karst. The treatment is applied of two variants, germinated seed in ascorbic acid (variantAand germinated seeds in disttilate water, than treated with ascorbic acid in 3 concentrations (variant B.

  1. Heterologous Expression and Characterization of an N-Acetyl-beta-D-hexosaminidase from Lactococcus lactis ssp. lactis IL1403

    Czech Academy of Sciences Publication Activity Database

    Nguyen, A. H.; Nguyen, T.-H.; Křen, Vladimír; Eijsink, V. G. H.; Haltrich, D.; Peterbauer, C.

    2012-01-01

    Roč. 60, č. 12 (2012), s. 3275-3281 ISSN 0021-8561 R&D Projects: GA ČR(CZ) GAP207/11/0629 Keywords : N-acetyl-beta-D-hexosaminidase * Lactococcus lactis ssp lactis IL1403 * pNP-GlcNAc Subject RIV: CE - Biochemistry Impact factor: 2.906, year: 2012

  2. Analysis of expressed sequence tags from Musa acuminata ssp. burmannicoides, var. Calcutta 4 (AA) leaves submitted to temperature stresses

    NARCIS (Netherlands)

    Santos, C.R.; Martins, N.F.; Horberg, H.M.; Almeida, E.R.P.; Coelho, M.C.F.; Togawa, R.; Silva, F.R.; Caetano, A.R.; Miller, R.N.G.; Souza, M.T.

    2005-01-01

    In order to discover genes expressed in leaves of Musa acuminata ssp. burmannicoides var. Calcutta 4 (AA), from plants submitted to temperature stress, we produced and characterized two full-length enriched cDNA libraries. Total RNA from plants subjected to temperatures ranging from 5°C to 25°C and

  3. Fate of Clavibacter michiganensis ssp. sepedonicus, the causal organism of bacterial ring rot in potato, in weeds and field crops.

    NARCIS (Netherlands)

    Wolf, van der J.M.; Beckhoven, van J.R.C.M.; Hukkanen, A.; Karjalainen, R.; Muller, P.

    2005-01-01

    Crops and weeds were tested for their ability to host Clavibacter michiganensis ssp. sepedonicus (Cms), the causal agent of bacterial ring rot in potato. Ten crops grown in rotation with potato in Europe, namely maize, wheat, barley, oat, bush bean, broad bean, rape, pea and onion and five cultivars

  4. Chemical composition and antifungal activities of essential oils of Satureja thymbra L. and Salvia pomifera ssp. calycina (Sm.) Hayek

    NARCIS (Netherlands)

    Glamoclija, J.; Sokovic, M.; Vukojevic, J.; Milenkovic, I.; Griensven, van L.J.L.D.

    2006-01-01

    This work covers the chemical composition and antifungal activities of essential oils isolated from savory (Satureja thymbra) and sage (Salvia pomifera ssp. calycina) analyzed using GC/MS. The main components of S. thymbra oil were gamma-terpinene (23.2%) and carvacrol (48.5%). The main components

  5. Lignan formation in hairy root cultures of Edelweiss (Leontopodium nivale ssp. alpinum (Cass.) Greuter)

    Science.gov (United States)

    Wawrosch, Christoph; Schwaiger, Stefan; Stuppner, Hermann; Kopp, Brigitte

    2014-01-01

    A hairy root line of Edelweiss (Leontopodium nivale ssp. alpinum (Cass.) Greuter) was obtained upon transformation with Agrobacterium rhizogenes strain ATCC15834. Elicitation of this line with silver nitrate, sucrose, methyl jasmonate and yeast extract at various concentrations in most cases resulted in a stimulation of lignan biosynthesis. Through elicitation with 6% sucrose the roots accumulated the pharmacologically active lignans leoligin and 5-methoxy-leoligin at levels of 0.0678% and 0.0372%, respectively, without significant growth inhibition. These lignan levels were comparable to those found in intact roots of cultivated Edelweiss. The biotechnological production of leoligin could be an attractive option for the continuous, field culture-independent production of the valuable secondary metabolites leoligin and 5-methoxy-leoligin. PMID:24932777

  6. Combining ability for maturity and plant height in brassica rapa (l.) ssp. dichotoma (roxb.) hanelt

    International Nuclear Information System (INIS)

    Nasim, A.; Farhatullah, A.; Khan, N.U.; Azam, S.M.; Nasim, Z.

    2014-01-01

    A 5 * 5 F1 diallel cross hybrids of Brassica rapa (L.) ssp. dichotoma (Roxb.) Hanelt along with parents were evaluated through combining ability for days to flowering (initiation and completion), days to maturity and plant height. Highly significant differences were recorded for all the traits. Mean squares due to general, specific and reciprocal combining ability were significant for all the traits except plant height for which the latter two components were non-significant. Prevalence of additive (plant height), non-additive (days to flowering completion; days to maturity) and reciprocal effects (days to flowering initiation) were detected. Parental line G-403 was best general combiner for all the traits. The F1 hybrids G-902 * G-265 (days to flowering initiation), G-902 * G-403 (days to flowering completion), G-265 * G-1500 (days to maturity) and G-909 * G-265 (plant height) were superior and may be exploited for future breeding programs. (author)

  7. Assessment of (Fouquieria splendens ssp. breviflora Cell Cultures Response Under to Water Stress

    Directory of Open Access Journals (Sweden)

    Leonor Angélica Guerrero Zúñiga

    2017-05-01

    Full Text Available Plant cell cultures are homogenous experimental systems, highly controllable that allow the study of short and large water stress adaptations without the interference of the different tissues and development of plants. An approach to understand these adaptations is through the presence of induced proteins; as a result of changes in genetic expression. This work analyze the response of Fouquieria splendens ssp. breviflora cell cultures exposed to abscisic acid (ABA, through the electrophoretic characterization of quantity and quality of stress induced proteins. There were recorded low molecular weight polypeptides (< 35kDa, common in experiments under ABA 10mM, followed by the association with 20 and 30mM ABA conditions, with a particularly response of cell cultures without the stress agent.

  8. Identification of Streptococcus equi ssp. zooepidemicus surface associated proteins by enzymatic shaving.

    Science.gov (United States)

    Wei, Zigong; Fu, Qiang; Liu, Xiaohong; Xiao, Pingping; Lu, Zhaohui; Chen, Yaosheng

    2012-10-12

    Streptococcus equi ssp. zooepidemicus (Streptococcus zooepidemicus, SEZ) is responsible for a wide variety of infections in many species. Attempts to control the infection caused by this agent are hampered by a lack of effective vaccines and useful diagnostic kits. Surface proteins of bacterial species are usually involved in interaction with host and hopefully act as biomarkers for serodiagnosis and subunit vaccine components. In this study, the surface proteins of SEZ C55138 strain were systematically identified by surface shaving with trypsin and a total of 20 surface associated proteins were found. Further analysis of five selected novel proteins (SzM, FBP, SAP, CSP and 5'-Nu) revealed that they all expressed in vivo and their recombinant derived proteins could be reactive with convalescent sera. These identified immunogenic surface proteins have potential as SEZ vaccine candidates and diagnostic markers. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. A new Epichloe species with interspecific hybrid origins from Poa pratensis ssp. pratensis in Liyang, China.

    Science.gov (United States)

    Yan, Kang; Yanling, Ji; Kunran, Zhu; Hui, Wang; Huimin, Miao; Zhiwei, Wang

    2011-01-01

    We describe a new Epichloë species found in symbiosis with Poa pratensis ssp. pratensis in Liyang, China. Stromata characteristic of Epichloë spp. were present on some of the reproductive tillers of individual host grasses. Only three of the 98 stromata observed on field plants became orange and produced perithecia. Phylogenetic analyses based on sequences of tubB and tefA indicated that this Epichloë sp. was an interspecific hybrid related to both E. yangzii and members in the E. typhina complex clade (ETC). Allele-1 of tefA and tubB grouped in the E. bromicola/E. yangzii clade; allele-2 of these two genes clustered in a distinct subclade in the ETC. This is the first report of an Epichloë species that has interspecific hybrid origins. We propose the name Epichloë liyangensis Z. Wang, Y. Kang et H. Miao, sp. nov. for this species.

  10. Óleo essencial de Gochnatia polymorpha (Less Cabr. ssp floccosa Cabr.

    Directory of Open Access Journals (Sweden)

    Maria Élida Alves Stefanello

    2006-10-01

    Full Text Available The essential oil isolated by hydrodistillation from flowers and root bark of Gochnatia polymorpha (Less Cabr. ssp floccosa Cabr. (Asteraceae was analyzed by capillary GC/MS. Thirty components were identified in oil of flowers. The composition depends on the stage of flowering. At the beginning of flowering, the main components were (E-nerolidol (20,4% and eugenol (17,9% whereas at the end they were phenylacetaldehyde (14,6% and tricosane (12,0%. In the essential oil of root bark ten sesquiterpenes were identified. The major components were beta-bisabolene (31,0% and beta-bisabolol (21,5%. This paper describes for the first time the composition of essential oil in the genus Gochnatia.

  11. Óleo essencial de Gochnatia polymorpha (Less Cabr. ssp floccosa Cabr.

    Directory of Open Access Journals (Sweden)

    Stefanello Maria Élida Alves

    2006-01-01

    Full Text Available The essential oil isolated by hydrodistillation from flowers and root bark of Gochnatia polymorpha (Less Cabr. ssp floccosa Cabr. (Asteraceae was analyzed by capillary GC/MS. Thirty components were identified in oil of flowers. The composition depends on the stage of flowering. At the beginning of flowering, the main components were (E-nerolidol (20,4% and eugenol (17,9% whereas at the end they were phenylacetaldehyde (14,6% and tricosane (12,0%. In the essential oil of root bark ten sesquiterpenes were identified. The major components were beta-bisabolene (31,0% and beta-bisabolol (21,5%. This paper describes for the first time the composition of essential oil in the genus Gochnatia.

  12. Reduction of linoleic acid inhibition in production of conjugated linoleic acid by Propionibacterium freudenreichii ssp. shermanii.

    Science.gov (United States)

    Rainio, A; Vahvaselkä, M; Suomalainen, T; Laakso, S

    2001-08-01

    A method for the production of conjugated linoleic acid (CLA) from linoleic acid (LA) using growing cultures of Propionibacterium freudenreichii ssp. shermanii JS was developed. The growth inhibitory effect of LA was eliminated by dispersing it in a sufficient concentration of polyoxyethylene sorbitan monooleate detergent. For the whey permeate medium used, the optimum LA:detergent ratio was 1:15 (w/w). As a result, the cultures tolerated at least 1000 microg x mL(-1) LA, which was converted to CLA with 57%-87% efficiency. The cis-9, trans-11 and trans-9, cis-11 isomers constituted 85%-90% of the CLA produced. The feasibility of the method was demonstrated also in de Man Rogosa-Sharpe (MRS) broth.

  13. Full scale test SSP 34m blade, combined load. Data report

    Energy Technology Data Exchange (ETDEWEB)

    Nielsen, Per H.; Nielsen, Magda; Jensen, Find M. (and others)

    2010-11-15

    This report is part of the research project where a 34m wind turbine blade from SSP-Technology A/S was tested in combined flap and edgewise load. The applied load is 55% of an imaginary extreme event based on the certification load of the blade. This report describes the reason for choosing the loads and the load direction and the method of applying the loads to the blade. A novel load introduction allows the blade to deform in a more realistic manner, allowing the observation of e.g. transverse shear distortion. The global and local deformation of the blade as well as the blades' respond to repeated tests has been studied and the result from these investigations are presented, including the measurements performed. (Author)

  14. Effect of explant origin and different growth regulators on micropropagation of Pistacia atlantica ssp. mutica

    Directory of Open Access Journals (Sweden)

    Ali-Ashraf Mehrabi

    2015-06-01

    Full Text Available Propagation of wild pistachio as a multipurpose woody species is a hard and tedious task. In this research, an effective in vitro protocol was developed for rapid proliferation of wild pistachio (Pistacia atlantica ssp. mutica in MS medium supplemented with B5 vitamins and different growth regulators. Rooting of plantlets was tested by two treatments containing Rhizopon and IBA in ex vitro. With respect to the results, the nodal segments explants, produced the highest shoot frequency, leaf frequency and the tallest shoots. On the other hand, the tallest shoots were generated from shoot tip explant and medium containing of TDZ plus IAA. Both treatments (Rhizopon and IBA led to a remarkable increase in the number of roots, root length and rooting percentage compared to the control. These results may be applied for rapid proliferation to spread the pistachio trees and shrubs that are difficult and time consuming.

  15. Assessing the diversity of arbuscular mycorrhizal fungi in semiarid shrublands dominated by Artemisia tridentata ssp. wyomingensis.

    Science.gov (United States)

    Carter, Keith A; Smith, James F; White, Merlin M; Serpe, Marcelo D

    2014-05-01

    Variation in the abiotic environment and host plant preferences can affect the composition of arbuscular mycorrhizal (AMF) assemblages. This study analyzed the AMF taxa present in soil and seedlings of Artemisia tridentata ssp. wyomingensis collected from sagebrush steppe communities in southwestern Idaho, USA. Our aims were to determine the AMF diversity within and among these communities and the extent to which preferential AMF-plant associations develop during seedling establishment. Mycorrhizae were identified using molecular methods following DNA extraction from field and pot culture samples. The extracted DNA was amplified using Glomeromycota specific primers, and identification of AMF was based on phylogenetic analysis of sequences from the large subunit-D2 rDNA region. The phylogenetic analyses revealed seven phylotypes, two within the Claroideoglomeraceae and five within the Glomeraceae. Four phylotypes clustered with known species including Claroideoglomus claroideum, Rhizophagus irregularis, Glomus microaggregatum, and Funneliformis mosseae. The other three phylotypes were similar to several published sequences not included in the phylogenetic analysis, but all of these were from uncultured and unnamed glomeromycetes. Pairwise distance analysis revealed some phylotypes with high genetic variation. The most diverse was the phylotype that included R. irregularis, which contained sequences showing pairwise differences up to 12 %. Most of the diversity in AMF sequences occurred within sites. The smaller genetic differentiation detected among sites was correlated with differences in soil texture. In addition, multiplication in pot cultures led to differentiation of AMF communities. Comparison of sequences obtained from the soil with those from A. tridentata roots revealed no significant differences between the AMF present in these samples. Overall, the sites sampled were dominated by cosmopolitan AMF taxa, and young seedlings of A. tridentata ssp

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

    Science.gov (United States)

    Hilborn, Elizabeth D.; Arduino, Matthew J.; Pruden, Amy; Edwards, Marc A.

    2015-01-01

    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 preexisting risk factors and frequently require hospitalization. Objectives The objectives of this report are to alert professionals of the impact of OPPPs, the fact that 30% of the population may be exposed to OPPPs, and the need to develop means to reduce OPPP exposure. We herein present a review of the epidemiology and ecology of these three bacterial OPPPs, specifically to identify common and unique features. Methods A Water Research Foundation–sponsored workshop gathered experts from across the United States to review the characteristics of OPPPs, identify problems, and develop a list of research priorities to address critical knowledge gaps with respect to increasing OPPP-associated disease. Discussion OPPPs share the common characteristics of disinfectant resistance and growth in biofilms in water distribution systems or premise plumbing. Thus, they share a number of habitats with humans (e.g., showers) that can lead to exposure and infection. The frequency of OPPP-infected individuals is rising and will likely continue to rise as the number of at-risk individuals is increasing. Improved reporting of OPPP disease and increased understanding of the genetic, physiologic, and structural characteristics governing the persistence and growth of OPPPs in drinking water distribution systems and premise plumbing is needed. Conclusions Because broadly effective community-level engineering interventions for the control of OPPPs have yet to be identified, and because the number of at-risk individuals will continue to rise, it is likely that OPPP-related infections will continue to increase. However, it is possible that individuals can take measures (e.g., raise hot water heater temperatures and filter

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

    Directory of Open Access Journals (Sweden)

    Patrícia Carvalho de Sequeira

    2005-11-01

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

  18. Comparison of Variable-Number Tandem-Repeat Markers typing and IS1245 Restriction Fragment Length Polymorphism fingerprinting of Mycobacterium avium subsp. hominissuis from human and porcine origins

    Directory of Open Access Journals (Sweden)

    Marttila Harri

    2010-03-01

    Full Text Available Abstract Background Animal mycobacterioses are regarded as a potential zoonotic risk and cause economical losses world wide. M. avium subsp. hominissuis is a slow-growing subspecies found in mycobacterial infected humans and pigs and therefore rapid and discriminatory typing methods are needed for epidemiological studies. The genetic similarity of M. avium subsp. hominissuis from human and porcine origins using two different typing methods have not been studied earlier. The objective of this study was to compare the IS1245 RFLP pattern and MIRU-VNTR typing to study the genetic relatedness of M. avium strains isolated from slaughter pigs and humans in Finland with regard to public health aspects. Methods A novel PCR-based genotyping method, variable number tandem repeat (VNTR typing of eight mycobacterial interspersed repetitive units (MIRUs, was evaluated for its ability to characterize Finnish Mycobacterium avium subsp. hominissuis strains isolated from pigs (n = 16 and humans (n = 13 and the results were compared with those obtained by the conventional IS1245 RFLP method. Results The MIRU-VNTR results showed a discriminatory index (DI of 0,92 and the IS1245 RFLP resulted in DI 0,98. The combined DI for both methods was 0,98. The MIRU-VNTR test has the advantages of being simple, reproducible, non-subjective, which makes it suitable for large-scale screening of M. avium strains. Conclusions Both typing methods demonstrated a high degree of similarity between the strains of human and porcine origin. The parallel application of the methods adds epidemiological value to the comparison of the strains and their origins. The present approach and results support the hypothesis that there is a common source of M. avium subsp. hominissuis infection for pigs and humans or alternatively one species may be the infective source to the other.

  19. Consequence of Mycobacterium avium complex pulmonary disease judging from the change of the chest CT image

    International Nuclear Information System (INIS)

    Fujiwara, Kiyohiro

    2008-01-01

    The long term consequence of the disease in Mycobacterium avium complex pulmonary disease (MACPD) is scarcely reported. This paper describes consequences of CT images and clinical symptoms in MACPD patients with rather poorer prognosis than usual during chemotherapy for one or more years in authors' hospital until May 2007. Subjects are 17 patients (average age 65.3 y, M 6/F 11) diagnosed as MACPD by the criteria by Jap. Soc. Tuberculosis (2003), whose follow up period is 14-105 (av. 58.1) months, and are classified in tuberculoid type (tt, 2 cases), bronchiectasis post surgery (2) and bronchia type (bt, 13, mostly primary MACPD). Chemotherapy is done with clarithromycin (CAM)+ethambutol (EB)+rifampicin (RHP) (+streptomycin (SM) for progression). Consequences of typical chest CT images are presented for each classification in this paper. Cavitation is seen even in bt as well as in tt and, if observed, the disease tends to deteriorate. In the secondary MACPD post surgery, the exacerbation of clinical symptom is often more severe despite slow changes in CT finding than in bt. Thus, careful follow up is necessary for the two cases above. (R.T.)

  20. Imaging features of Mycobacterium avium-intracellulare complex (MAC) in children with AIDS

    Energy Technology Data Exchange (ETDEWEB)

    Pursner, M. [State University of New York, Health Science Center at Brooklyn, New York, NY (United States); Haller, J.O. [Beth Israel Medical Center, Department of Radiology, New York, NY (United States); Berdon, W.E. [Babies Hospital, Department of Radiology, New York, NY (United States)

    2000-06-01

    Purpose. The purpose of this paper was to review the imaging features of Mycobacterium avium-intracellulare complex (MAC) in 16 pediatric patients with human immunodeficiency virus (HIV). Materials and methods. We reviewed the pertinent clinical records of 16 children diagnosed with MAC between January 1990 and June 1998. These 16 cases were blood- or biopsy-proven to have MAC infection. Their plain films, abdominal, and chest CT scans were then reviewed and the findings were analyzed with reference to the few reported cases of children with MAC. Results. Abdominal findings: all but one had retroperitoneal adenopathy, mesenteric adenopathy or both. Ten patients had hepatomegaly, while nine patients were found to have splenomegaly. Four patients had nonspecific thickened gallbladder wall, while intestinal wall thickening and thickened stomach folds were identified in six of ten patients. Necrotic, fluid-filled nodes were also found. Chest findings included mediastinal adenopathy, cystic/cavitary lesions and bronchiectasis. One patient developed a fistula between the mediastinal lymph nodes, esophagus, and bronchial tree. Conclusion. Pediatric patients with HIV who develop MAC infection may present with massive lymph-node enlargement. This can occur not only in mesenteric and retroperitoneal nodes but also in hilar and posterior mediastinal nodes as well. As in MTB infection, these nodes can break down with development of fistulous tracts to both esophagus and adjacent lung. The major differential diagnostic consideration besides MTB is lymphoma. (orig.)