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Sample records for glabrata experimentally infected

  1. Competitive Interactions between C. albicans, C. glabrata and C. krusei during Biofilm Formation and Development of Experimental Candidiasis

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    Rossoni, Rodnei Dennis; Barbosa, Júnia Oliveira; Vilela, Simone Furgeri Godinho; dos Santos, Jéssica Diane; de Barros, Patrícia Pimentel; Prata, Márcia Cristina de Azevedo; Anbinder, Ana Lia; Fuchs, Beth Burgwyn; Jorge, Antonio Olavo Cardoso; Mylonakis, Eleftherios; Junqueira, Juliana Campos

    2015-01-01

    In this study, we evaluated the interactions between Candida albicans, Candida krusei and Candida glabrata in mixed infections. Initially, these interactions were studied in biofilms formed in vitro. CFU/mL values of C. albicans were lower in mixed biofilms when compared to the single biofilms, verifying 77% and 89% of C. albicans reduction when this species was associated with C. glabrata and C. krusei, respectively. After that, we expanded this study for in vivo host models of experimental candidiasis. G. mellonella larvae were inoculated with monotypic and heterotypic Candida suspensions for analysis of survival rate and quantification of fungal cells in the haemolymph. In the groups with single infections, 100% of the larvae died within 18 h after infection with C. albicans. However, interaction groups achieved 100% mortality after 72 h of infection by C. albicans-C. glabrata and 96 h of infection by C. albicans-C. krusei. C. albicans CFU/mL values from larvae hemolymph were lower in the interacting groups compared with the monoespecies group after 12 h of infection. In addition, immunosuppressed mice were also inoculated with monotypic and heterotypic microbial suspensions to induce oral candidiasis. C. albicans CFU/mL values recovered from oral cavity of mice were higher in the group with single infection by C. albicans than the groups with mixed infections by C. albicans-C. glabrata and C. albicans-C. krusei. Moreover, the group with single infection by C. albicans had a higher degree of hyphae and epithelial changes in the tongue dorsum than the groups with mixed infections. We concluded that single infections by C. albicans were more harmful for animal models than mixed infections with non-albicans species, suggesting that C. albicans establish competitive interactions with C. krusei and C. glabrata during biofilm formation and development of experimental candidiasis. PMID:26146832

  2. Competitive Interactions between C. albicans, C. glabrata and C. krusei during Biofilm Formation and Development of Experimental Candidiasis.

    Science.gov (United States)

    Rossoni, Rodnei Dennis; Barbosa, Júnia Oliveira; Vilela, Simone Furgeri Godinho; dos Santos, Jéssica Diane; de Barros, Patrícia Pimentel; Prata, Márcia Cristina de Azevedo; Anbinder, Ana Lia; Fuchs, Beth Burgwyn; Jorge, Antonio Olavo Cardoso; Mylonakis, Eleftherios; Junqueira, Juliana Campos

    2015-01-01

    In this study, we evaluated the interactions between Candida albicans, Candida krusei and Candida glabrata in mixed infections. Initially, these interactions were studied in biofilms formed in vitro. CFU/mL values of C. albicans were lower in mixed biofilms when compared to the single biofilms, verifying 77% and 89% of C. albicans reduction when this species was associated with C. glabrata and C. krusei, respectively. After that, we expanded this study for in vivo host models of experimental candidiasis. G. mellonella larvae were inoculated with monotypic and heterotypic Candida suspensions for analysis of survival rate and quantification of fungal cells in the haemolymph. In the groups with single infections, 100% of the larvae died within 18 h after infection with C. albicans. However, interaction groups achieved 100% mortality after 72 h of infection by C. albicans-C. glabrata and 96 h of infection by C. albicans-C. krusei. C. albicans CFU/mL values from larvae hemolymph were lower in the interacting groups compared with the monoespecies group after 12 h of infection. In addition, immunosuppressed mice were also inoculated with monotypic and heterotypic microbial suspensions to induce oral candidiasis. C. albicans CFU/mL values recovered from oral cavity of mice were higher in the group with single infection by C. albicans than the groups with mixed infections by C. albicans-C. glabrata and C. albicans-C. krusei. Moreover, the group with single infection by C. albicans had a higher degree of hyphae and epithelial changes in the tongue dorsum than the groups with mixed infections. We concluded that single infections by C. albicans were more harmful for animal models than mixed infections with non-albicans species, suggesting that C. albicans establish competitive interactions with C. krusei and C. glabrata during biofilm formation and development of experimental candidiasis.

  3. Failed Reverse Total Shoulder Arthroplasty Caused by Recurrent Candida glabrata Infection with Prior Serratia marcescens Coinfection

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    John G. Skedros

    2014-01-01

    Full Text Available This report describes a 58-year-old insulin-dependent diabetic male patient who initially sustained a proximal humerus fracture from a fall. The fracture fixation failed and then was converted to a humeral hemiarthroplasty, which became infected with Candida glabrata and Serratia marcescens. After these infections were believed to be cured with antibacterial and antifungal treatments and two-stage irrigation and debridement, he underwent conversion to a reverse total shoulder arthroplasty. Unfortunately, the C. glabrata infection recurred and, nearly 1.5 years after implantation of the reverse total shoulder, he had a resection arthroplasty (removal of all implants and cement. His surgical and pharmacologic treatment concluded with (1 placement of a tobramycin-impregnated cement spacer also loaded with amphotericin B, with no plan for revision arthroplasty (i.e., the spacer was chronically retained, and (2 chronic use of daily oral fluconazole. We located only three reported cases of Candida species causing infection in shoulder arthroplasties (two C. albicans, one C. parapsilosis. To our knowledge, a total shoulder arthroplasty infected with C. glabrata has not been reported, nor has a case of a C. glabrata and S. marcescens periprosthetic coinfection in any joint. In addition, it is well known that S. marcescens infections are uncommon in periprosthetic joint infections.

  4. Candida glabrata: an emerging oral opportunistic pathogen.

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    Li, L; Redding, S; Dongari-Bagtzoglou, A

    2007-03-01

    Following the widespread use of immunosuppressive therapy and broad-spectrum antimycotic prophylaxis, C. glabrata has emerged as an important opportunistic pathogen in the oral mucosa. In the past, studies on the virulence factors and host-pathogen interactions of this organism were scarce, but continued to rise in recent years. Denture-wearing, immunosuppression, antibiotic therapy, and aging are risk factors for oral colonization or infection with C. glabrata. Compared with C. albicans, C. glabrata exhibits lower oral keratinocyte-adherence capacity, but higher denture-surface-adherence ability. The role of extracellular hydrolase production in the virulence of this organism does not appear to be as important as it is in C. albicans pathogenesis. Although traditionally thought of as a non-transforming yeast organism, both phenotypic switching and pseudohyphal formation have recently been identified in C. glabrata, but their role in pathogenesis is not known. With the exception of granulocyte monocyte colony-stimulating factor, C. glabrata triggers a lower proinflammatory cytokine response in oral epithelial cells than does C. albicans, in a strain-dependent manner. C. glabrata is less susceptible to killing by human beta-defensins than is C. albicans and exhibits various degrees of resistance to the antifungal activity of salivary histatins and mucins. In addition, C. glabrata possesses both innate and acquired resistance against antifungal drugs, due to its ability to modify ergosterol biosynthesis, mitochondrial function, or antifungal efflux. This resistance allows for its relative overgrowth over other susceptible species and may contribute to the recent emergence of C. glabrata infections in chronically immunocompromised populations. Further investigations on the virulence and host-pathogen interactions of C. glabrata are needed to better define the pathogenesis of oral C. glabrata infection in susceptible hosts.

  5. Candida glabrata Binding to Candida albicans Hyphae Enables Its Development in Oropharyngeal Candidiasis.

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    Tati, Swetha; Davidow, Peter; McCall, Andrew; Hwang-Wong, Elizabeth; Rojas, Isolde G; Cormack, Brendan; Edgerton, Mira

    2016-03-01

    Pathogenic mechanisms of Candida glabrata in oral candidiasis, especially because of its inability to form hyphae, are understudied. Since both Candida albicans and C. glabrata are frequently co-isolated in oropharyngeal candidiasis (OPC), we examined their co-adhesion in vitro and observed adhesion of C. glabrata only to C. albicans hyphae microscopically. Mice were infected sublingually with C. albicans or C. glabrata individually, or with both species concurrently, to study their ability to cause OPC. Infection with C. glabrata alone resulted in negligible infection of tongues; however, colonization by C. glabrata was increased by co-infection or a pre-established infection with C. albicans. Furthermore, C. glabrata required C. albicans for colonization of tongues, since decreasing C. albicans burden with fluconazole also reduced C. glabrata. C. albicans hyphal wall adhesins Als1 and Als3 were important for in vitro adhesion of C. glabrata and to establish OPC. C. glabrata cell wall protein coding genes EPA8, EPA19, AWP2, AWP7, and CAGL0F00181 were implicated in mediating adhesion to C. albicans hyphae and remarkably, their expression was induced by incubation with germinated C. albicans. Thus, we found a near essential requirement for the presence of C. albicans for both initial colonization and establishment of OPC infection by C. glabrata.

  6. A Murine Model of Candida glabrata Vaginitis Shows No Evidence of an Inflammatory Immunopathogenic Response.

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    Evelyn E Nash

    Full Text Available Candida glabrata is the second most common organism isolated from women with vulvovaginal candidiasis (VVC, particularly in women with uncontrolled diabetes mellitus. However, mechanisms involved in the pathogenesis of C. glabrata-associated VVC are unknown and have not been studied at any depth in animal models. The objective of this study was to evaluate host responses to infection following efforts to optimize a murine model of C. glabrata VVC. For this, various designs were evaluated for consistent experimental vaginal colonization (i.e., type 1 and type 2 diabetic mice, exogenous estrogen, varying inocula, and co-infection with C. albicans. Upon model optimization, vaginal fungal burden and polymorphonuclear neutrophil (PMN recruitment were assessed longitudinally over 21 days post-inoculation, together with vaginal concentrations of IL-1β, S100A8 alarmin, lactate dehydrogenase (LDH, and in vivo biofilm formation. Consistent and sustained vaginal colonization with C. glabrata was achieved in estrogenized streptozotocin-induced type 1 diabetic mice. Vaginal PMN infiltration was consistently low, with IL-1β, S100A8, and LDH concentrations similar to uninoculated mice. Biofilm formation was not detected in vivo, and co-infection with C. albicans did not induce synergistic immunopathogenic effects. This data suggests that experimental vaginal colonization of C. glabrata is not associated with an inflammatory immunopathogenic response or biofilm formation.

  7. A Murine Model of Candida glabrata Vaginitis Shows No Evidence of an Inflammatory Immunopathogenic Response.

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    Nash, Evelyn E; Peters, Brian M; Lilly, Elizabeth A; Noverr, Mairi C; Fidel, Paul L

    2016-01-01

    Candida glabrata is the second most common organism isolated from women with vulvovaginal candidiasis (VVC), particularly in women with uncontrolled diabetes mellitus. However, mechanisms involved in the pathogenesis of C. glabrata-associated VVC are unknown and have not been studied at any depth in animal models. The objective of this study was to evaluate host responses to infection following efforts to optimize a murine model of C. glabrata VVC. For this, various designs were evaluated for consistent experimental vaginal colonization (i.e., type 1 and type 2 diabetic mice, exogenous estrogen, varying inocula, and co-infection with C. albicans). Upon model optimization, vaginal fungal burden and polymorphonuclear neutrophil (PMN) recruitment were assessed longitudinally over 21 days post-inoculation, together with vaginal concentrations of IL-1β, S100A8 alarmin, lactate dehydrogenase (LDH), and in vivo biofilm formation. Consistent and sustained vaginal colonization with C. glabrata was achieved in estrogenized streptozotocin-induced type 1 diabetic mice. Vaginal PMN infiltration was consistently low, with IL-1β, S100A8, and LDH concentrations similar to uninoculated mice. Biofilm formation was not detected in vivo, and co-infection with C. albicans did not induce synergistic immunopathogenic effects. This data suggests that experimental vaginal colonization of C. glabrata is not associated with an inflammatory immunopathogenic response or biofilm formation.

  8. Influence of the nontarget mollusc Marisa cornuarietis on the hourly cercarial production of Schistosoma mansoni from Biomphalaria glabrata.

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    Ferrer, J R; Pointier, J P; Théron, A; Moné, H

    1991-10-01

    A comparative study of hourly cercarial productivities of Schistosoma mansoni from infected Biomphalaria glabrata was carried out in the presence of either healthy B. glabrata (control) or healthy Marisa cornuarietis (experimental). The results showed that, with M. cornuarietis, almost all the hourly cercarial productivities increased by a factor varying from 1.3 to 2.5 without modification of the shedding period.

  9. Distribution and Schistosoma mansoni infection of Biomphalaria glabrata in different habitats in a rural area in the Jequitinhonha Valley, Minas Gerais, Brazil: environmental and epidemiological aspects

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    Helmut Kloos

    2004-11-01

    Full Text Available This paper examines the distribution and infection of Biomphalaria glabrata with Schistosoma mansoni in all aquatic snail habitats in a rural area in the state of Minas Gerais, Brazil, in relation to physico/biotic and behavioral factors. Snail and environmental surveys were carried out semi-annually between July 2001 and November 2002 at 106 sites. Collected snails were examined in the laboratory for infection. B. glabrata densities were highest in overflow ponds, irrigation ponds, springs, canals and wells, and lowest in fishponds and water tanks. Snail densities were higher during the hot, rainy season except for streams and canals and were statistically associated with the presence of fish, pollution, and vegetation density. Tilapia fish and an unidentified Diptera larva were found to be predators of B. glabrata but ducks were not. Twenty-four of the 25 infected snails were collected in 2001(1.4% infection rate and only one in 2002, after mass chemotherapy. The occurrence of B. glabrata in all 11 snail habitats both at and away from water contact sites studied indicates widespread risk of human infection in the study area. In spite of the strong association between B. glabrata and tilapia in fishponds we do not recommend its use in schistosomiasis control for ecological reasons and its relative inefficiency in streams and dams.

  10. Generational distribution of a Candida glabrata population: Resilient old cells prevail, while younger cells dominate in the vulnerable host.

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    Bouklas, Tejas; Alonso-Crisóstomo, Luz; Székely, Tamás; Diago-Navarro, Elizabeth; Orner, Erika P; Smith, Kalie; Munshi, Mansa A; Del Poeta, Maurizio; Balázsi, Gábor; Fries, Bettina C

    2017-05-01

    Similar to other yeasts, the human pathogen Candida glabrata ages when it undergoes asymmetric, finite cell divisions, which determines its replicative lifespan. We sought to investigate if and how aging changes resilience of C. glabrata populations in the host environment. Our data demonstrate that old C. glabrata are more resistant to hydrogen peroxide and neutrophil killing, whereas young cells adhere better to epithelial cell layers. Consequently, virulence of old compared to younger C. glabrata cells is enhanced in the Galleria mellonella infection model. Electron microscopy images of old C. glabrata cells indicate a marked increase in cell wall thickness. Comparison of transcriptomes of old and young C. glabrata cells reveals differential regulation of ergosterol and Hog pathway associated genes as well as adhesion proteins, and suggests that aging is accompanied by remodeling of the fungal cell wall. Biochemical analysis supports this conclusion as older cells exhibit a qualitatively different lipid composition, leading to the observed increased emergence of fluconazole resistance when grown in the presence of fluconazole selection pressure. Older C. glabrata cells accumulate during murine and human infection, which is statistically unlikely without very strong selection. Therefore, we tested the hypothesis that neutrophils constitute the predominant selection pressure in vivo. When we altered experimentally the selection pressure by antibody-mediated removal of neutrophils, we observed a significantly younger pathogen population in mice. Mathematical modeling confirmed that differential selection of older cells is sufficient to cause the observed demographic shift in the fungal population. Hence our data support the concept that pathogenesis is affected by the generational age distribution of the infecting C. glabrata population in a host. We conclude that replicative aging constitutes an emerging trait, which is selected by the host and may even play an

  11. Assimilation of NAD(+) precursors in Candida glabrata.

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    Ma, Biao; Pan, Shih-Jung; Zupancic, Margaret L; Cormack, Brendan P

    2007-10-01

    The yeast pathogen Candida glabrata is a nicotinamide adenine dinucleotide (NAD(+)) auxotroph and its growth depends on the environmental supply of vitamin precursors of NAD(+). C. glabrata salvage pathways defined in this article allow NAD(+) to be synthesized from three compounds - nicotinic acid (NA), nicotinamide (NAM) and nicotinamide riboside (NR). NA is salvaged through a functional Preiss-Handler pathway. NAM is first converted to NA by nicotinamidase and then salvaged by the Preiss-Handler pathway. Salvage of NR in C. glabrata occurs via two routes. The first, in which NR is phosphorylated by the NR kinase Nrk1, is independent of the Preiss-Handler pathway. The second is a novel pathway in which NR is degraded by the nucleosidases Pnp1 and Urh1, with a minor role for Meu1, and ultimately converted to NAD(+) via the nicotinamidase Pnc1 and the Preiss-Handler pathway. Using C. glabrata mutants whose growth depends exclusively on the external NA or NR supply, we also show that C. glabrata utilizes NR and to a lesser extent NA as NAD(+) sources during disseminated infection.

  12. Proteomics of drug resistance in Candida glabrata biofilms.

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    Seneviratne, C Jayampath; Wang, Yu; Jin, Lijian; Abiko, Y; Samaranayake, Lakshman P

    2010-04-01

    Candida glabrata is a fungal pathogen that causes a variety of mucosal and systemic infections among compromised patient populations with higher mortality rates. Previous studies have shown that biofilm mode of the growth of the fungus is highly resistant to antifungal agents compared with the free-floating or planktonic mode of growth. Therefore, in the present study, we used 2-D DIGE to evaluate the differential proteomic profiles of C. glabrata under planktonic and biofilm modes of growth. Candida glabrata biofilms were developed on polystyrene surfaces and age-matched planktonic cultures were obtained in parallel. Initially, biofilm architecture, viability, and antifungal susceptibility were evaluated. Differentially expressed proteins more than 1.5-fold in DIGE analysis were subjected to MS/MS. The transcriptomic regulation of these biomarkers was evaluated by quantitative real-time PCR. Candida glabrata biofilms were highly resistant to the antifungals and biocides compared with the planktonic mode of growth. Candida glabrata biofilm proteome when compared with its planktonic proteome showed upregulation of stress response proteins, while glycolysis enzymes were downregulated. Similar trend could be observed at transcriptomic level. In conclusion, C. glabrata biofilms possess higher amount of stress response proteins, which may potentially contribute to the higher antifungal resistance seen in C. glabrata biofilms.

  13. Candida glabrata Esophagitis: Are We Seeing the Emergence of a New Azole-Resistant Pathogen?

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    Aze Wilson

    2014-01-01

    Full Text Available Background. Candida glabrata (C. glabrata has become a recognized pathogen in fungal esophagitis. A proportion of these isolates are azole-resistant which may have treatment implications. Variability in the prevalence of this organism exists in the limited data available. Objective. To determine the incidence of C. glabrata esophagitis in a North American hospital setting and to highlight factors that may predispose patients to this condition. Methods. Patient charts were collected from January 1, 2009 to July 30, 2011. Any charts of patients identified as having esophagitis with a positive fungal culture were reviewed for the species of Candida and the presence of factors that would predispose them to esophageal candidiasis. Results. The prevalence of Candida esophagitis based on culture was 2.2% (37 subjects. C. glabrata was the 2nd most prevalent pathogen identified (24.3% or 9 subjects. Of the C. glabrata cohort, all patients had at least one factor predisposing them to candidiasis. Conclusion. C. glabrata esophagitis makes up a large portion of the candidal esophagitis seen in hospital. C. glabrata infections were associated with at least one risk factor for candidal infection. Given its resistance to azole-based therapy, this may have treatment implications for how candidal esophagitis is approached by the clinician.

  14. Candida glabrata olecranon bursitis treated with bursectomy and intravenous caspofungin.

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    Skedros, John G; Keenan, Kendra E; Trachtenberg, Joel D

    2013-01-01

    Orthopedic surgeons are becoming more involved in the care of patients with septic arthritis and bursitis caused by yeast species. This case report involves a middle-aged immunocompromised female who developed a Candida glabrata septic olecranon bursitis that developed after she received a corticosteroid injection in the olecranon bursa for presumed aseptic bursitis. Candida (Torulopsis) glabrata is the second most frequently isolated Candida species from the bloodstream in the United States. Increased use of fluconazole and other azole antifungal agents as a prophylactic treatment for recurrent Candida albicans infections in immunocompromised individuals is one reason why there appears to be increased resistance of C. glabrata and other nonalbicans Candida (NAC) species to fluconazole. In this patient, this infection was treated with surgery (bursectomy) and intravenous caspofungin, an echinocandin. This rare infectious etiology coupled with this intravenous antifungal treatment makes this case novel among cases of olecranon bursitis caused by yeasts.

  15. Human schistosomiasis in Puerto Rico: reduced prevalence rate and absence of Biomphalaria glabrata.

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    Giboda, M; Malek, E A; Correa, R

    1997-11-01

    A combined epidemiologic and malacologic survey of schistosomiasis in Puerto Rico was carried out in areas where previous surveys had reported the prevalence of the disease. This limited survey, with 495 persons examined, found a low prevalence (0.6%) of Schistosoma mansoni infections. The infections were restricted to three people more than 36 years of age. No infections were detected in children 16 years of age or less, and this cohort comprised 57.8% of the study group. Malacologic surveys of the four streams, 10 rivers, and eight lakes throughout the island revealed the absence of intermediate host Biomphalaria glabrata and the presence of Thiara granifera, a competitive species of B. glabrata and the predatory snail Marisa cornuarietis. We believe that the absence of B. glabrata is the primary reason for the sustained reduction in the prevalence of schistosomiasis in Puerto Rico.

  16. Osteomyelitis Caused by Candida glabrata in the Distal Phalanx

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    Shunichi Toki

    2014-01-01

    Full Text Available Osteomyelitis caused by Candida glabrata is rare and its optimal treatment is unknown. Here we report a case of osteomyelitis caused by C. glabrata in the distal phalanx in a 54-year-old woman. Despite partial resection of the nail and administering a 1-month course of antibiotics for paronychia, the local swelling remained and an osteolytic lesion was found. C. glabrata osteomyelitis of the distal phalanx was later diagnosed after curettage. Thereafter, the patient was treated with antifungal agents for 3 months. The infection eventually resolved, and radiological healing of the osteolytic lesion was achieved. Antifungal susceptibility testing should be performed in the case of osteomyelitis caused by nonalbicans Candida species, due to their resistance to fluconazole.

  17. Candida glabrata prosthetic joint infection, successfully treated with anidulafungin: A case report and review of the literature.

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    Koutserimpas, Christos; Samonis, George; Velivassakis, Emmanouil; Iliopoulou-Kosmadaki, Stylliani; Kontakis, Georgios; Kofteridis, Diamantis P

    2018-04-01

    Non-albicans Candida prosthetic joint infection (PJI) is extremely rare. A case of a Candida glabrata knee PJI is a 68-year-old splenectomised female smoker, suffering from chronic obstructive pulmonary disease (COPD) and alcoholism is reported. The patient presented with a peri-prosthetic fracture, 15 years after total knee replacement surgery. Cultures of the intraoperative peri-prosthetic tissue and materials yielded C. glabrata, as well as a methicillin-resistant S. epidermitis. The patient was treated with anidulafungin and vancomycin. The knee prosthetic joint was removed and cement-spacer with vancomycin and gentamycin was placed. Additionally, an external fixation was performed. A second stage revision surgery was planned, after completion of the antimicrobial and antifungal treatment. The patient is followed up for 4 months without signs, symptoms or findings of infection. PJI Candida infections require a high clinical suspicion index. It is of utmost importance to report these cases, since there is no consensus yet of the proper antifungal treatment. Furthermore, a literature review regarding treatment of those cases is provided. First-line treatment with an echinocandin seems most proper, due to their fungicidal properties, their effectiveness against biofilm, as well as their minimal toxicity, making them ideal for long-term use. Further experience is needed, for better understanding the disease's pathogenesis and optimal treatment. © 2017 Blackwell Verlag GmbH.

  18. Time-kill assay and Etest evaluation for synergy with polymyxin B and fluconazole against Candida glabrata.

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    Pankey, George; Ashcraft, Deborah; Kahn, Heather; Ismail, Abdulrahim

    2014-10-01

    Fluconazole-resistant Candida glabrata is an emerging pathogen that causes fungemia. Polymyxin B, a last-resort antibiotic used to treat multidrug-resistant Gram-negative bacterial infections, has been found to possess in vitro fungicidal activity and showed synergy with fluconazole against a single strain of C. glabrata. Since both agents may be used simultaneously in intensive care unit (ICU) patients, this study was performed to test for possible synergy of this combination against 35 C. glabrata blood isolates, using 2 methods: a time-kill assay and an experimental MIC-MIC Etest method. Thirty-five genetically unique C. glabrata bloodstream isolates were collected from 2009 to 2011, identified using an API 20C system, and genotyped by repetitive sequence-based PCR (rep-PCR). MICs were determined by Etest and broth microdilution methods. Synergy testing was performed using a modified bacterial Etest synergy method and time-kill assay, with final results read at 24 h. The Etest method showed synergy against 19/35 (54%) isolates; the time-kill assay showed synergy against 21/35 (60%) isolates. Isolates not showing drug synergy had an indifferent status. Concordance between methods was 60%. In vitro synergy of polymyxin B and fluconazole against the majority of C. glabrata isolates was demonstrated by both methods. The bacterial Etest synergy method adapted well when used with C. glabrata. Etest was easier to perform than time-kill assay and may be found to be an acceptable alternative to time-kill assay with antifungals. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  19. Candida glabrata Biofilms: How Far Have We Come?

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    Célia F. Rodrigues

    2017-03-01

    Full Text Available Infections caused by Candida species have been increasing in the last decades and can result in local or systemic infections, with high morbidity and mortality. After Candida albicans, Candida glabrata is one of the most prevalent pathogenic fungi in humans. In addition to the high antifungal drugs resistance and inability to form hyphae or secret hydrolases, C. glabrata retain many virulence factors that contribute to its extreme aggressiveness and result in a low therapeutic response and serious recurrent candidiasis, particularly biofilm formation ability. For their extraordinary organization, especially regarding the complex structure of the matrix, biofilms are very resistant to antifungal treatments. Thus, new approaches to the treatment of C. glabrata’s biofilms are emerging. In this article, the knowledge available on C. glabrata’s resistance will be highlighted, with a special focus on biofilms, as well as new therapeutic alternatives to control them.

  20. Candida glabrata Biofilms: How Far Have We Come?

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    Rodrigues, Célia F.; Rodrigues, Maria Elisa; Silva, Sónia; Henriques, Mariana

    2017-01-01

    Infections caused by Candida species have been increasing in the last decades and can result in local or systemic infections, with high morbidity and mortality. After Candida albicans, Candida glabrata is one of the most prevalent pathogenic fungi in humans. In addition to the high antifungal drugs resistance and inability to form hyphae or secret hydrolases, C. glabrata retain many virulence factors that contribute to its extreme aggressiveness and result in a low therapeutic response and serious recurrent candidiasis, particularly biofilm formation ability. For their extraordinary organization, especially regarding the complex structure of the matrix, biofilms are very resistant to antifungal treatments. Thus, new approaches to the treatment of C. glabrata’s biofilms are emerging. In this article, the knowledge available on C. glabrata’s resistance will be highlighted, with a special focus on biofilms, as well as new therapeutic alternatives to control them. PMID:29371530

  1. [Evaluation of a rapid trehalase test for the identification of Candida glabrata].

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    Kirdar, Sevin; Gültekin, Berna; Evcil, Gonca; Ozkütük, Aydan; Sener, Asli Gamze; Aydin, Neriman

    2009-04-01

    Candida species which cause local infections, may also lead to fatal systemic infections. The increasing incidence of non-albicans Candida, especially fluconazole susceptible or resistant dose-dependent C. glabrata, increased the importance of rapid and accurate species level identification for Candida. Rapid and correct identification of C. glabrata is essential for the initiation of the appropriate antifungal therapy. This study was conducted to evaluate the performance of the rapid trehalase test in the diagnosis of C. glabrata isolates. A total of 173 Candida strains isolated from various clinical specimens and identified according to germ tube test, growth on cornmeal Tween 80 agar and the colony morphologies on Mast-CHROMagar Candida medium (Mast Diagnostics, UK), were included to the study. The identification of non-albicans Candida species were also confirmed by API 20CAUX (BioMerieux, France) system. Accordingly 86 (50%) of the isolates were identified as C. glabrata, 48 (28%) C. albicans, 17 (10%) C. krusei, 13 (8%) C. tropicalis, 5 (3%) C. parapsilosis, 3 (2%) C. kefyr and 1 (1%) Cutilis. In order to detect the presence of trehalase enzyme in Condida strains, all isolates were grown on Sabouraud dextrose agar containing 4% glucose and then one yeast colony was emulsified in 50 microl of citrate buffer containing 4% (wt/vol) trehalose for 3 h at 37 degrees C. Presence of glucose which emerged after the action of trehalase on trehalose, was detected by a commercial "urinary glucose detection dipstick" (Spinreacta, Spain). All C. glabrata strains yielded positive result by trehalase test. None C. glabrata isolates were found negative by trehalase test except for one strain of C. tropicalis. In this study, the trehalase test allowed identification of C. globrata with 100% sensitivity and 98.9% specificity. It was concluded that trehalase test is a rapid, cost-effective and simple test that can be used for the accurate identification of C. glabrata.

  2. Oxidative stress response to menadione and cumene hydroperoxide in the opportunistic fungal pathogen Candida glabrata

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    Mayra Cuéllar-Cruz

    2009-07-01

    Full Text Available Candida glabrata is an opportunistic fungal pathogen that can cause severe invasive infections and can evade phagocytic cell clearance. We are interested in understanding the virulence of this fungal pathogen, in particular its oxidative stress response. Here we investigated C. glabrata, Saccharomyces cerevisiae and Candida albicans responses to two different oxidants: menadione and cumene hydroperoxide (CHP. In log-phase, in the presence of menadione, C. glabrata requires Cta1p (catalase, while in a stationary phase (SP, Cta1p is dispensable. In addition, C. glabrata is less resistant to menadione than C. albicans in SP. The S. cerevisiae laboratory reference strain is less resistant to menadione than C. glabrata and C. albicans; however S. cerevisiaeclinical isolates (CIs are more resistant than the lab reference strain. Furthermore, S. cerevisiae CIs showed an increased catalase activity. Interestingly, in SP C. glabrata and S. cerevisiae are more resistant to CHP than C. albicans and Cta1p plays no apparent role in detoxifying this oxidant.

  3. MALDI-TOF typing highlights geographical and fluconazole resistance clusters in Candida glabrata.

    Science.gov (United States)

    Dhieb, C; Normand, A C; Al-Yasiri, M; Chaker, E; El Euch, D; Vranckx, K; Hendrickx, M; Sadfi, N; Piarroux, R; Ranque, S

    2015-06-01

    Utilizing matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectra for Candida glabrata typing would be a cost-effective and easy-to-use alternative to classical DNA-based typing methods. This study aimed to use MALDI-TOF for the typing of C. glabrata clinical isolates from various geographical origins and test its capacity to differentiate between fluconazole-sensitive and -resistant strains.Both microsatellite length polymorphism (MLP) and MALDI-TOF mass spectra of 58 C. glabrata isolates originating from Marseilles (France) and Tunis (Tunisia) as well as collection strains from diverse geographic origins were analyzed. The same analysis was conducted on a subset of C. glabrata isolates that were either susceptible (MIC ≤ 8 mg/l) or resistant (MIC ≥ 64 mg/l) to fluconazole.According to the seminal results, both MALDI-TOF and MLP classifications could highlight C. glabrata population structures associated with either geographical dispersal barriers (p typing to investigate C. glabrata infection outbreaks and predict the antifungal susceptibility profile of clinical laboratory isolates. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  4. Heteroresistance to Fluconazole Is a Continuously Distributed Phenotype among Candida glabrata Clinical Strains Associated with In Vivo Persistence

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    Ronen Ben-Ami

    2016-08-01

    Full Text Available Candida glabrata causes persistent infections in patients treated with fluconazole and often acquires resistance following exposure to the drug. Here we found that clinical strains of C. glabrata exhibit cell-to-cell variation in drug response (heteroresistance. We used population analysis profiling (PAP to assess fluconazole heteroresistance (FLCHR and to ask if it is a binary trait or a continuous phenotype. Thirty (57.6% of 52 fluconazole-sensitive clinical C. glabrata isolates met accepted dichotomous criteria for FLCHR. However, quantitative grading of FLCHR by using the area under the PAP curve (AUC revealed a continuous distribution across a wide range of values, suggesting that all isolates exhibit some degree of heteroresistance. The AUC correlated with rhodamine 6G efflux and was associated with upregulation of the CDR1 and PDH1 genes, encoding ATP-binding cassette (ABC transmembrane transporters, implying that HetR populations exhibit higher levels of drug efflux. Highly FLCHRC. glabrata was recovered more frequently than nonheteroresistant C. glabrata from hematogenously infected immunocompetent mice following treatment with high-dose fluconazole (45.8% versus 15%, P = 0.029. Phylogenetic analysis revealed some phenotypic clustering but also variations in FLCHR within clonal groups, suggesting both genetic and epigenetic determinants of heteroresistance. Collectively, these results establish heteroresistance to fluconazole as a graded phenotype associated with ABC transporter upregulation and fluconazole efflux. Heteroresistance may explain the propensity of C. glabrata for persistent infection and the emergence of breakthrough resistance to fluconazole.

  5. On the origin of the Biomphalaria glabrata hemocytes

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    Samaly dos Santos Souza

    2006-10-01

    Full Text Available A histologic, morphometric and ultrastructural study performed on Biomphalaria glabrata submitted to infection with Schistosoma mansoni miracidia failed to provide significant evidences that the so-called amebocyte-producing organ (APO is really the central organ for hemocyte production. In infected snails no general reactive changes appeared in the APO, the mitoses were seen only occasionally, and the possibility of cellular hyperplasia was ruled out by morphometric measurements. Under the electron microscope the APO cells presented an essentially epithelial structure, without features indicative of transition toward hemocytes. On the other hand, the present findings pointed to a multicentric origin for the mollusck hemocytes, as earlier studies had indicated. Dense foci of hemocyte collections appeared sometimes around disintegrating sporocysts and cercariae in several organs and tissues of the infected snails, including a curious accumulation of such cells inside the ventricular cavity of the heart. In the heart and other sites, features suggestive of transformation of vascular space endothelial lining cells into hemocytes were apparent. To some extent, the postulated multicentric origin for B. glabrata hemocytes recapitulates earlier embryologic findings in vertebrates, when mesenchymal vascular spaces generate the circulating and phagocytic blood cells.

  6. Probiotic yeast Saccharomyces boulardii (nom. nud.) modulates adhesive properties of Candida glabrata.

    Science.gov (United States)

    Tomičić, Zorica; Zupan, Jure; Matos, Tadeja; Raspor, Peter

    2016-11-01

    Following the widespread use of immunosuppressive therapy together with broad-spectrum antimycotic therapy, the frequency of mucosal and systemic infections caused by the pathogenic yeast Candida glabrata has increased in the past decades. Due to the resistance of C. glabrata to existing azole drugs, it is very important to look for new strategies helping the treatment of such fungal diseases. In this study, we investigated the effect of the probiotic yeast Saccharomyces boulardii (nom. nud.) on C. glabrata adhesion at different temperatures, pH values, and in the presence of fluconazole, itraconazole and amphotericin B. We also studied the adhesion of C. glabrata co-culture with Candida krusei, Saccharomyces cerevisiae, two bacterial probiotics Lactobacillus rhamnosus and Lactobacillus casei The method used to assess adhesion was crystal violet staining. Our results showed that despite the nonadhesiveness of S. boulardii cells, this probiotic significantly affected the adherence ability of C. glabrata This effect was highly dependent on C. glabrata strain and was either antagonistic or synergistic. Regarding the extrinsic factors, temperature did not indicate any significant influence on this S. boulardii modulatory effect, while at high pH and at increased concentrations of antimycotics, S. boulardii did not manage to repress the adhesion of C. glabrata strains. The experiments of C. glabrata co-cultures with other species showed that the adhesiveness of two separate cultures could not be used to predict the adhesiveness of their co-culture. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. Relative Abundances of Candida albicans and Candida glabrata in In Vitro Coculture Biofilms Impact Biofilm Structure and Formation.

    Science.gov (United States)

    Olson, Michelle L; Jayaraman, Arul; Kao, Katy C

    2018-04-15

    Candida is a member of the normal human microbiota and often resides on mucosal surfaces such as the oral cavity or the gastrointestinal tract. In addition to their commensality, Candida species can opportunistically become pathogenic if the host microbiota is disrupted or if the host immune system becomes compromised. An important factor for Candida pathogenesis is its ability to form biofilm communities. The two most medically important species- Candida albicans and Candida glabrata -are often coisolated from infection sites, suggesting the importance of Candida coculture biofilms. In this work, we report that biofilm formation of the coculture population depends on the relative ratio of starting cell concentrations of C. albicans and C. glabrata When using a starting ratio of C. albicans to C. glabrata of 1:3, ∼6.5- and ∼2.5-fold increases in biofilm biomass were observed relative to those of a C. albicans monoculture and a C. albicans / C. glabrata ratio of 1:1, respectively. Confocal microscopy analysis revealed the heterogeneity and complex structures composed of long C. albicans hyphae and C. glabrata cell clusters in the coculture biofilms, and reverse transcription-quantitative PCR (qRT-PCR) studies showed increases in the relative expression of the HWP1 and ALS3 adhesion genes in the C. albicans / C. glabrata 1:3 biofilm compared to that in the C. albicans monoculture biofilm. Additionally, only the 1:3 C. albicans / C. glabrata biofilm demonstrated an increased resistance to the antifungal drug caspofungin. Overall, the results suggest that interspecific interactions between these two fungal pathogens increase biofilm formation and virulence-related gene expression in a coculture composition-dependent manner. IMPORTANCE Candida albicans and Candida glabrata are often coisolated during infection, and the occurrence of coisolation increases with increasing inflammation, suggesting possible synergistic interactions between the two Candida species in

  8. Female biased sex-ratio in Schistosoma mansoni after exposure to an allopatric intermediate host strain of Biomphalaria glabrata.

    Science.gov (United States)

    Lepesant, Julie M J; Boissier, Jérôme; Climent, Déborah; Cosseau, Céline; Grunau, Christoph

    2013-10-01

    For parasites that require multiple hosts to complete their development, the interaction with the intermediate host may have an impact on parasite transmission and development in the definitive host. The human parasite Schistosoma mansoni needs two different hosts to complete its life cycle: the freshwater snail Biomphalaria glabrata (in South America) as intermediate host and a human or rodents as final host. To investigate the influence of the host environment on life history traits in the absence of selection, we performed experimental infections of two B. glabrata strains of different geographic origin with the same clonal population of S. mansoni. One B. glabrata strain is the sympatric host and the other one the allopatric host. We measured prevalence in the snail, the cercarial infectivity, sex-ratio, immunopathology in the final host and microsatellite frequencies of individual larvae in three successive generations. We show that, even if the parasite population is clonal based on neutral markers, S. mansoni keeps the capacity of generating phenotypic plasticity and/or variability for different life history traits when confront to an unusual environment, in this study the intermediate host. The most dramatic change was observed in sex-ratio: in average 1.7 times more female cercariae were produced when the parasite developed in an allopatric intermediate host. Copyright © 2013 Elsevier Inc. All rights reserved.

  9. The Effectiveness of Voriconazole in Therapy of Candida glabrata's Biofilms Oral Infections and Its Influence on the Matrix Composition and Gene Expression.

    Science.gov (United States)

    Rodrigues, Célia F; Gonçalves, Bruna; Rodrigues, Maria Elisa; Silva, Sónia; Azeredo, Joana; Henriques, Mariana

    2017-08-01

    Candida glabrata is one of most prevalent yeast in fungal infections, especially in immunocompromised patients. Its azole resistance results in a low therapeutic response, particularly when associated with biofilms. The main goal of this work was to study the effectiveness of voriconazole (Vcz) against C. glabrata biofilms oral pathologies, as esophageal or oropharyngeal candidiasis. Antifungal susceptibilities were determined in pre-formed 24-h-biofilms and ERG genes expression was determined by qRT-PCR. Protein quantification was performed using BCA ® Kit, carbohydrate was estimated according to the Dubois assay and β-1,3 glucans concentration were determined using Glucatell ® kit. Finally, ergosterol, Vcz, and fluconazole (Flu) concentrations within the biofilm matrices were determined by RP-HPLC. Results showed that C. glabrata biofilms were more susceptible to Vcz than to Flu and that ERG genes expression evidenced an overexpression of the three ERG genes in the presence of both azoles. The matrix content presented a remarked decrease in proteins and an increase in carbohydrates, namely β-1,3 glucans. Ergosterol was successfully detected and quantified in the biofilm matrices, with no differences in all the considered conditions. Vcz demonstrated better diffusion through the biofilms and better cell penetration capacities, than Flu, indicating that the structure of the drug molecule fully influences its dissemination through the biofilm matrices. This work showed that Vcz is notably more effective than Flu for the treatment of resistant C. glabrata oral biofilms, which demonstrates a clinical relevance in its future use for the treatment of oropharyngeal/esophageal candidiasis caused by this species.

  10. Estudo quantitativo de metais presentes na hemolinfa de Biomphalaria glabrata (Gastropoda, infectadas e não infectadas com Schistosoma mansoni Quantitative study of metal present in the hemolymph of Biomphalaria glabrata (Gastropoda, infected and uninfected with Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Marco Antonio Vasconcelos Santos

    2005-04-01

    Full Text Available Inicialmente, desenvolveu-se um estudo para quantificar e comparar as concentrações de alguns metais presentes em duas amostras de hemolinfa do caramujo Biomphalaria glabrata (infectados e não-infectados com Schistosoma mansoni. A espectrometria de emissão óptica com fonte de plasma induzido (ICP-OES, foi utilizada para analisar os metais nas duas amostras. Os metais estudados foram: alumínio, cálcio, cádmio, cobalto, cromo, cobre, ferro, potássio, magnésio, manganês, chumbo e zinco. Os resultados mostram que, a princípio, os metais não são fatores determinantes no processo de defesa desses organismos contra este parasita, quando presente nos seus tecidos.We conducted a preliminary study to quantify and compare two concentrations of the same metals present in the hemolymph of snail Biomphalaria glabrata. In this context, we used Induction Coupled Plasma Optical Emission Spectroscopy technique (ICP-OES, to analyze the metals in the two samples (snails infected and not infected with Schistosoma mansoni. The metals studied were: aluminum, calcium, cadmium, cobalt, chromium, copper, iron, potassium, magnesium, manganese, lead and zinc. Preliminary results showed that such metals are not involved in the defense of these organisms against the parasite, when present in their tissues.

  11. Mini-chromosomes among danish Candida glabrata isolates originated through two different mechanisms

    DEFF Research Database (Denmark)

    Ahmad, K. M.; Ishchuk, O.; Hellborg, L.

    2012-01-01

    We analyzed 201 strains of the pathogenic yeast Candida glabrata from patients, mainly suffering from systemic infection, at Danish hospitals during 1985 – 1999. Our analysis showed that these strains were closely related but exhibited large karyotype polymorphism. Nine strains contained mini-chr...

  12. Development of Schistosoma mansoni in Biomphalaria tenagophila, Biomphalaria straminea and Biomphalaria glabrata Desenvolvimento do Schistosoma mansoni em Biomphalaria tenagophila, Biomphalaria straminea e Biomphalaria glabrata

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    Cecilia Pereira de Souza

    1995-06-01

    Full Text Available A comparative study of the development of Schistosoma mansoni during the intra-molluscan phase was made by means of histological sections of Biomphalaria tenagophila, B. straminea and B. glabrata from Brazil. Two hundred snails of each species were individually exposed to 50 miracidia of the S. mansoni, AL line. No larvae were observed in the snails fixed 72 h after exposure. In specimens shedding cercariae, 31 days after exposure tissue reactions encapsulating the larvae were seen in B. tenagophila and B. straminea, in the head-foot, mantle collar and renal ducts. No tissue reactions occurred in the digestive glands of these two species. In B. glabrata the presence of numerous sporocysts and cercariae without tissue reactions was observed in the digestive gland, and other organs. The levels of infection of the snails and the average numbers of cercariae shed per day were 32.6% and 79±90 respectively for B. tenagophila, 11.3% and 112±100 for B. straminea and 75.3% and 432±436 for B. glabrata. The lower levels of infection and average numbers of cercariae shed by B. tenagophila and B. straminea are thus related to their more potent internal defense systems.Foi feito estudo comparativo do desenvolvimento do Schistosoma mansoni na fase intra-molusco, através de cortes histológicos, em Biomphalaria tenagophila, B. straminea e B. glabrata. Duzentos moluscos de cada espécie foram expostos individualmente a 50 miracídios de S. mansoni da linhagem AL. Nenhuma larva foi observada nos exemplares fixados 72 horas após a exposição. Nos exemplares eliminando cercárías, 31 dias após a exposição, foram observadas reações teciduais de encapsulamento de larvas em B. tenagophila e B. straminea, na região cefalopodal, colar do manto e dutos renais. Nas glândulas digestivas das duas espécies não foram observadas reações. Em B. glabrata foi registrada a presença de numerosos esporocistos e cercárias sem reação tecidual na gl

  13. Behavior of Schistosoma mansoni-induced histopathological lesions in Biomphalaria glabrata submitted to ionizing radiation

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    Azevedo Carine M.

    2004-01-01

    Full Text Available Present report demonstrates that repeated radiation of Schistosoma mansoni-infected Biomphalaria glabrata, totaling 15,000 rads, caused a sudden, albeit transient, suppression of cercarial shedding. Initially, sporocysts practically disappeared from the snail tissues. The more resistant developing cercariae presented nuclear clumping and vacuolation, before undergoing lysis. No host tissue reaction was evident at any time. Thirty-four days after the last irradiation, the snails resumed cercarial elimination. By that time numerous sporocysts and developing cercariae were detected, disseminated throughout snail tissues in a pattern similar to that of a highly malignant neoplasm, with no signs of host cellular reactions, which on the other hand were present in non-irradiated infected controls. The region of the ovo-testis was apparently destroyed after radiation, but returned to its normal appearance around 40 days after the last radiation. Ionizing radiation affected both host and parasite in S. mansoni-infected Biomphalaria glabrata, but the resulting impressive changes were soon reversed.

  14. Activity of Polyphenolic Compounds against Candida glabrata

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    Ricardo Salazar-Aranda

    2015-09-01

    Full Text Available Opportunistic mycoses increase the morbidity and mortality of immuno-compromised patients. Five Candida species have been shown to be responsible for 97% of worldwide cases of invasive candidiasis. Resistance of C. glabrata and C. krusei to azoles has been reported, and new, improved antifungal agents are needed. The current study was designed to evaluatethe activity of various polyphenolic compounds against Candida species. Antifungal activity was evaluated following the M27-A3 protocol of the Clinical and Laboratory Standards Institute, and antioxidant activity was determined using the DPPH assay. Myricetin and baicalein inhibited the growth of all species tested. This effect was strongest against C. glabrata, for which the minimum inhibitory concentration (MIC value was lower than that of fluconazole. The MIC values against C. glabrata for myricitrin, luteolin, quercetin, 3-hydroxyflavone, and fisetin were similar to that of fluconazole. The antioxidant activity of all compounds was confirmed, and polyphenolic compounds with antioxidant activity had the greatest activity against C. glabrata. The structure and position of their hydroxyl groups appear to influence their activity against C. glabrata.

  15. Epithelial GM-CSF induction by Candida glabrata.

    Science.gov (United States)

    Li, L; Dongari-Bagtzoglou, A

    2009-08-01

    The main cytokine induced by the interaction of oral epithelial cells with C. glabrata is granulocyte monocyte colony-stimulating factor (GM-CSF); however, the mechanisms regulating this response are unknown. Based on previously published information on the interactions of C. albicans with oral epithelial cells, we hypothesized that interaction with viable C. glabrata triggers GM-CSF synthesis via NF-kappaB activation. We found that C. glabrata-induced GM-CSF synthesis was adhesion-dependent, enhanced by endocytosis, and required fungal viability. NF-kappaB activation was noted during interaction of epithelial cells with C. glabrata, and pre-treatment with an NF-kappaB inhibitor partly inhibited GM-CSF synthesis. Blocking TLR4 with anti-TLR4 antibody did not inhibit GM-CSF production. In contrast, an anti-CDw17 antibody triggered significant inhibition of NF-kappaB activation and GM-CSF synthesis. beta-glucans did not stimulate GM-CSF synthesis, suggesting that the CDw17/NF-kappaB/GM-CSF pathway may be beta-glucan-independent. This study provides new insights into the mechanism of GM-CSF induction by C. glabrata.

  16. One small step for a yeast--microevolution within macrophages renders Candida glabrata hypervirulent due to a single point mutation.

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    Sascha Brunke

    2014-10-01

    Full Text Available Candida glabrata is one of the most common causes of candidemia, a life-threatening, systemic fungal infection, and is surpassed in frequency only by Candida albicans. Major factors contributing to the success of this opportunistic pathogen include its ability to readily acquire resistance to antifungals and to colonize and adapt to many different niches in the human body. Here we addressed the flexibility and adaptability of C. glabrata during interaction with macrophages with a serial passage approach. Continuous co-incubation of C. glabrata with a murine macrophage cell line for over six months resulted in a striking alteration in fungal morphology: The growth form changed from typical spherical yeasts to pseudohyphae-like structures - a phenotype which was stable over several generations without any selective pressure. Transmission electron microscopy and FACS analyses showed that the filamentous-like morphology was accompanied by changes in cell wall architecture. This altered growth form permitted faster escape from macrophages and increased damage of macrophages. In addition, the evolved strain (Evo showed transiently increased virulence in a systemic mouse infection model, which correlated with increased organ-specific fungal burden and inflammatory response (TNFα and IL-6 in the brain. Similarly, the Evo mutant significantly increased TNFα production in the brain on day 2, which is mirrored in macrophages confronted with the Evo mutant, but not with the parental wild type. Whole genome sequencing of the Evo strain, genetic analyses, targeted gene disruption and a reverse microevolution experiment revealed a single nucleotide exchange in the chitin synthase-encoding CHS2 gene as the sole basis for this phenotypic alteration. A targeted CHS2 mutant with the same SNP showed similar phenotypes as the Evo strain under all experimental conditions tested. These results indicate that microevolutionary processes in host-simulative conditions

  17. Acute extrarenal kidney damage in the course of infection with fungal strain of Candida glabrata in a patient with type 2 diabetes

    International Nuclear Information System (INIS)

    Szarejko-Paradowska, A.; Bartnicki, P.; Pietrzak, B.; Wilk, R.; Serwa-Stepien, E.; Rysz, J.; Jablonowski, Z.

    2010-01-01

    Background: Acute renal injury is becoming a significant epidemiological problem among patients requiring hospital treatment. Extrarenal aetiology of the kidney injury is recognized in 5 % to 10 % of hospitalized patients; however, the identification of the mycelium of the Candida glabrata as the direct factor causing the acute urinary obstruction is extremely rare. Case Report: A 64-year-old woman was admitted to the clinic because of progressing weakness, nausea and vomiting, poor appetite and reduced urination. On admission, laboratory findings revealed pyuria, inflammatory changes, acute renal failure (eGFR-MDRD 6 ml/min), and hyperglycemia. The patient underwent USG of the abdominal cavity, which showed bilateral hydronephrosis, with lithiasis on the right site. Cystoscopy done the next day revealed that the mucous membrane of the bladder was reddened and had a white coating. During the next several days, a renal fistula was created on the left and right sides. Candida glabrata was isolated from urine, and was sensitive only to voriconazole. V-fend (voriconazole) treatment resulted in increase of diuresis and decrease in creatinine and urea levels. Conclusions: Urinary tract infection caused by Candida glabrata causes significant therapeutic problems. In most cases, these yeasts are resistant to triazole anti-fungal drugs such as fluconazole, which translates into significantly increased mortality of patients. To date, a similar case was described only by one group of doctors, however, due to the intensity of the currently used immunosuppression and multiantibiotic therapy, increased incidence of diabetes and the aging of the population, it is expected that the prevalence of this clinical problem will increase. (authors)

  18. Candida species isolated from different body sites and their antifungal susceptibility pattern: Cross-analysis of Candida albicans and Candida glabrata biofilms.

    Science.gov (United States)

    Cataldi, Valentina; Di Campli, Emanuela; Fazii, Paolo; Traini, Tonino; Cellini, Luigina; Di Giulio, Mara

    2017-08-01

    Candida species are regular commensal in humans, but-especially in immunocompromised patients-they represent opportunistic pathogens giving rise to systemic infection. The aim of the present work was to isolate and characterize for their antifungal profile Candida species from different body sites and to analyze the biofilms produced by C. albicans and C. glabrata isolates. Eighty-one strains of Candida species from 77 patients were identified. Epidemiological study showed that the most isolated species were C. albicans (44), C. glabrata (13) and C. parapsilosis (13) mainly from Hematology, Infectious Diseases, Medicine, Neonatology and Oncology Divisions, the majority of the biological samples were swabs (44) and blood cultures (16). The analysis of the biofilm formation was performed at 24 and 48-hours comparing resistant and susceptible strains of C. albicans to resistant and susceptible strains of C. glabrata. Candida albicans has a greater ability to form biofilm compared to C. glabrata, both in the susceptible and resistant strains reaching maturity after 24 hours with a complex structure composed of blastospores, pseudohyphae, and hyphae embedded in a matrix. On the contrary, C. glabrata biofilm was composed exclusively of blastospores that in the resistant strain, after 24 hours, were organized in a compact multilayer different to the discontinuous structure observed in the susceptible analyzed strains. In conclusion, the increasing of the incidence of Candida species infection together with their emerging drug resistance also related to the biofilm forming capability underline the need to monitor their distribution and susceptibility patterns for improving the surveillance and for a correct management of the infection. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  19. Discriminative power of fatty acid methyl ester (FAME) analysis using the microbial identification system (MIS) for Candida (Torulopsis) glabrata and Saccharomyces cerevisiae.

    Science.gov (United States)

    Peltroche-Llacsahuanga, H; Schmidt, S; Lütticken, R; Haase, G

    2000-12-01

    Candida (Torulopsis) glabrata is frequently isolated in cases of fungal infection and commonly shows acquired or innate fluconazole resistance. Saccharomyces cerevisiae, an emerging opportunistic yeast pathogen, causes serious systemic infections in immunocompromised, and vaginitis and superficial infections in immunocompetent patients. For both species reliable identification in the routine laboratory is mandatory, but species identification of strains, e.g. trehalose-negative C. glabrata, may be difficult. Therefore, gas-liquid chromatography (GLC) of whole cell fatty acid methyl ester (FAME) profiles, that is independent of assimilation profiles of strains and suitable for reliable and rapid identification of clinically important yeasts, was applied. However, frequent misidentification of C. glabrata as S. cerevisiae has been reported when using the Yeast Clinical Database of MIS. Accuracy of MIS identification may be strongly influenced by the amounts of cell mass analyzed. Therefore, the present study compared the MIS results of these two yeasts achieved with different cell masses. Primarily we optimized, especially with respect to cost-effectiveness, the recommended streaking technique yielding a maximal recovery of 90-130 mg of cell mass from one plate, enabling testing of poor growing strains of C. glabrata. For all C. glabrata strains tested (n = 10) the highest identification scores (SI [Similarity Index] range 0.525-0.963, median 0.832) were achieved with 30 to 45 mg of cell mass. Only 5 of 10 S. cerevisiae strains revealed good library comparisons (SI > or = 0.5) when using 30 mg of cell mass, whereas with 45 mg all strains but two revealed this SI-level. For S. cerevisiae a higher amount of cell mass processed (up to 90 mg) was correlated with better identification scores (SI range using 90 mg: 0.464-0.870, median, 0.737). Several passages prior to FAME analysis of C. glabrata strains on recommended media revealed narrowing of SI ranges, but

  20. Analgesic effect of leaf extract from Ageratina glabrata in the hot plate test

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    Guadalupe García P

    2011-10-01

    Full Text Available Ageratina glabrata (Kunth R.M. King & H. Rob., Asteraceae (syn. Eupatorium glabratum Kunth is widely distributed throughout Mexico and popularly known as "chamizo blanco" and "hierba del golpe" for its traditional use as external analgesic remedy. Though glabrata species has been chemically studied, there are no experimentally asserted reports about possible analgesic effects which can be inferred from its genus Ageratina. To fill the gap, we evaluated A. glabrata extracts in an animal model of nociception exploiting thermal stimuli. NMR and mass analyses identified a new thymol derivative, 10-benzoiloxy-6,8,9-trihydroxy-thymol isobutyrate (1, which was computationally converted into a ring-closed structure to explain interaction with the COX-2 enzyme in a ligand-receptor docking study. The resulting docked pose is in line with reported crystal complexes of COX-2 with chromene ligands. Based on the present results of dichloromethane extracts from its dried leaves, it is safe to utter that the plant possesses analgesic effects in animal tests which are mediated through inhibition of COX-2 enzyme.

  1. Molluscicidal and ovicidal activities of plant extracts of the Piperaceae on Biomphalaria glabrata (Say, 1818).

    Science.gov (United States)

    Rapado, L N; Nakano, E; Ohlweiler, F P; Kato, M J; Yamaguchi, L F; Pereira, C A B; Kawano, T

    2011-03-01

    Schistosomiasis is a tropical disease caused by Schistosoma and occurs in 54 countries, mainly in South America, the Caribbean region, Africa and the eastern Mediterranean. Currently, 5 to 6 million Brazilian people are infected and 30,000 are under infection risk. Typical of poor regions, this disease is associated with the lack of basic sanitation and very frequently to the use of contaminated water in agriculture, housework and leisure. One of the most efficient methods of controlling the disease is application of molluscicides to eliminate or to reduce the population of the intermediate host snail Biomphalaria glabrata. Studies on molluscicidal activity of plant extracts have been stimulated by issues such as environmental preservation, high cost and recurrent resistance of snails to synthetic molluscicides. The aim of this study was to determine the molluscicide action of extracts from Piperaceae species on adult and embryonic stages of B. glabrata. Fifteen extracts from 13 Piperaceae species were obtained from stems, leaves and roots. Toxicity of extracts was evaluated against snails at two different concentrations (500 and 100 ppm) and those causing 100% mortality at 100 ppm concentration were selected to obtain the LC₉₀ (lethal concentration of 90% mortality). Piper aduncum, P. crassinervium, P. cuyabanum, P. diospyrifolium and P. hostmannianum gave 100% mortality of adult snails at concentrations ranging from 10 to 60 ppm. These extracts were also assayed on embryonic stages of B. glabrata and those from P. cuyabanum and P. hostmannianum showed 100% ovicidal action at 20 ppm.

  2. Carboxymethylated ɩ-carrageenan conjugated amphotericin B loaded gelatin nanoparticles for treating intracellular Candida glabrata infections.

    Science.gov (United States)

    Aparna, V; Melge, Anu Rohit; Rajan, V K; Biswas, Raja; Jayakumar, R; Gopi Mohan, C

    2018-04-15

    Intercellular Candida glabrata infections are difficult to treat due to poor penetration of drugs into the fungal niche. Delivering amphotericin B (Amp B) into the macrophages where the pathogen inhabits is an effective solution. We are studying the macrophage targeting proficiency of ɩ-carrageenan for the delivery of Amp B using gelatin A nanoparticles (GNPs). The choice of gelatin A was the outcome of in silico inspections where the amino functionalized polymer having the best docking score with Amp B was selected. We prepared a sustained release formulation of amp B loaded carboxymethyl ɩ-carrageenan conjugated gelatin nanoparticles (CMC-Amp B-GNPs) with size 343±12nm and -25±5.3mV zeta potential. The formulations were found to be stable, biocompatible and non-haemolytic. Flow cytometry analysis showed 3 fold higher uptake of CMC-GNPs compared to the GNPs by RAW 264.7 cells. CMC-Amp B-GNPs showed enhanced antifungal activity than bare Amp B and Amp B-GNPs. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. The mating type-like loci of Candida glabrata.

    Science.gov (United States)

    Yáñez-Carrillo, Patricia; Robledo-Márquez, Karina A; Ramírez-Zavaleta, Candy Y; De Las Peñas, Alejandro; Castaño, Irene

    2014-01-01

    Candida glabrata, a haploid and opportunistic fungal pathogen that has not known sexual cycle, has conserved the majority of the genes required for mating and cell type identity. The C. glabrata genome contains three mating-type-like loci called MTL1, MTL2 and MTL3. The three loci encode putative transcription factors, a1, α1 and α2 that regulate cell type identity and sexual reproduction in other fungi like the closely related Saccharomyces cerevisiae. MTL1 can contain either a or α information. MTL2, which contains a information and MTL3 with α information, are relatively close to two telomeres. MTL1 and MTL2 are transcriptionally active, while MTL3 is subject to an incomplete silencing nucleated at the telomere that depends on the silencing proteins Sir2, Sir3, Sir4, yKu70/80, Rif1, Rap1 and Sum1. C. glabrata does not seem to maintain cell type identity, as cell type-specific genes are expressed regardless of the type (or even absence) of mating information. These data highlight important differences in the control of mating and cell type identity between the non-pathogenic yeast S. cerevisiae and C. glabrata, which might explain the absence of a sexual cycle in C. glabrata. The fact that C. glabrata has conserved the vast majority of the genes involved in mating might suggest that some of these genes perhaps have been rewired to control other processes important for the survival inside the host as a commensal or as a human pathogen. This manuscript is part of the series of works presented at the "V International Workshop: Molecular genetic approaches to the study of human pathogenic fungi" (Oaxaca, Mexico, 2012). Copyright © 2013 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.

  4. Overexpression of Aldo-Keto-Reductase in Azole-resistant Clinical Isolates of Candida Glabrata Determined by cDNA-AFLP

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    Mansour Heidari

    2013-01-01

    Full Text Available Background: Candida glabrata causes significant medical problems in immunocompromised patients. Many strains of this yeast are intrinsically resistant to azole antifungal agents, and treatment is problematic, leading to high morbidity and mortality rates in immunosuppressed individuals. The primary goal of this study was to investigate the genes involved in the drug resistance of clinical isolates of C. glabrata.Methods: The clinical isolates of C. glabrata were collected in an epidemiological survey of candidal infection inimmunocompromised patients and consisted of four fluconazole and itraconazole resistant isolates, two fluconazoleand itraconazole sensitive isolates, and C. glabrata CBS 138 as reference strain. Antifungal susceptibility patterns ofthe organisms were determined beforehand by the Clinical and Laboratory Standards Institute (CLSI. The potentialgene(s implicated in antifungal resistance were investigated using complementary DNA- Amplified Fragment Length Polymorphism (cDNA-AFLP. Semi-quantitative RT-PCR was carried out to evaluate the expression of gene(s in resistant isolates as compared to sensitive and reference strains.Results and conclusions: The aldo-keto-reductase superfamily (AKR gene was upregulated in the resistant clinicalisolates as assessed by cDNA-AFLP. Semi-quantitative RT-PCR revealed AKR mRNA expression approximately twice that seen in the sensitive isolates. Overexpression of the AKR gene was associated with increased fluconazole and itraconazole resistance in C. glabrata. The data suggest that upregulation of the AKR gene might give a new insight into the mechanism of azole resistance.

  5. Synergistic effects of tacrolimus and azole antifungal compounds in fluconazole-susceptible and fluconazole-resistant Candida glabrata isolates

    Directory of Open Access Journals (Sweden)

    Laura Bedin Denardi

    2015-03-01

    Full Text Available In vitro interaction between tacrolimus (FK506 and four azoles (fluconazole, ketoconazole, itraconazole and voriconazole against thirty clinical isolates of both fluconazole susceptible and -resistant Candida glabrata were evaluated by the checkerboard microdilution method. Synergistic, indifferent or antagonism interactions were found for combinations of the antifungal agents and FK506. A larger synergistic effect was observed for the combinations of FK506 with itraconazole and voriconazole (43%, followed by that of the combination with ketoconazole (37%, against fluconazole-susceptible isolates. For fluconazole-resistant C. glabrata, a higher synergistic effect was obtained from FK506 combined with ketoconazole (77%, itraconazole (73%, voriconazole (63% and fluconazole (60%. The synergisms that we observed in vitro, notably against fluconazole-resistant C. glabrata isolates, are promising and warrant further analysis of their applications in experimental in vivo studies.

  6. Synergistic anticandidal activity of menthol in combination with itraconazole and nystatin against clinical Candida glabrata and Candida krusei isolates.

    Science.gov (United States)

    Sharifzadeh, Aghil; Khosravi, Ali Reza; Shokri, Hojjatollah; Tari, Paria Samadi

    2017-06-01

    Candida glabrata (C. glabrata) and C. krusei are now emerging as serious hospital acquired infections in immunocompromised patients. Menthol, a terpenic compound, has been reported to have antifungal activity. The aim of this study was to investigate the effect of menthol in combination with itraconazole or nystatin against C. glabrata and C. krusei isolates. The effects of menthol along with itraconazole and nystatin, were evaluated by the Clinical Laboratory Standards Institute (CLSI) M44-A and CLSI M27-A3 methods. The fractional inhibitory concentration index (FICI) was determined for menthol plus itraconazole and nystatin combinations using the checkerboard method. The mean of minimum inhibitory concentration (MIC) values of menthol, nystatin and itraconazole were 53.2, 2.30 and 1.50 μg/ml for C. glabrata isolates and 121, 1.08 and 0.38 μg/ml for C. krusei isolates, respectively. Menthol in combination with itraconazole or nystatin exhibited the synergistic effects against all species of Candida tested. FICI values for menthol plus itraconazole and nystatin combinations ranged from 0.250 to 0.561 and 0.139 to 0.623 for C. glabrata isolates, and 0.182 to 0.750 and 0.188 to 0.760 for C. krusei, respectively. These results support the potential use of menthol as an anticandidal agent, and it can be used complementarily with other conventional antifungal agents. Copyright © 2017. Published by Elsevier Ltd.

  7. Gastrointestinal transit and digestive rhythm in Biomphalaria glabrata (Say)

    International Nuclear Information System (INIS)

    Florschutz, A.; Becker, W.

    1999-01-01

    Transit of X-ray opaque Micropaque™ particles in the digestive tract of Biomphalaria glabrata was investigated by X-radiography. Passage through the whole animal lasted up to 200 min depending on the experimental conditions (starvation, fed ad libitum etc.). For the first time, cyclic activity of the digestive gland could be visualised: the gland is emptied every 60-110 min discharging undigested particles. In the afternoon transport of probably predigested material from more distally located parts of the digestive tract into the esophagus can be observed indicating resorption processes occurring in the esophagus

  8. Candida glabrata um patogénio emergente?

    OpenAIRE

    Matilde, Filipa Alexandra Veiga

    2014-01-01

    Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz Nas últimas décadas as infecções fúngicas têm aumentado exponencialmente. Apesar de Candida albicans (C. albicans) continuar a ser a espécie isolada com maior frequência, as espécies não albicans tem vindo a proliferar rapidamente. Candida glabrata (C. glabrata) é uma das espécies emergentes como patogénio oportunista humano, sendo responsável maioritariamente por candidoses invasivas em pacie...

  9. Effects of high dose rate gamma radiation on survival and reproduction of Biomphalaria glabrata

    Energy Technology Data Exchange (ETDEWEB)

    Cantinha, Rebeca S.; Nakano, Eliana [Instituto Butantan, Sao Paulo, SP (Brazil). Lab. de Parasitologia], e-mail: rebecanuclear@gmail.com, e-mail: eliananakano@butantan.gov.br; Borrely, Sueli I. [Instituto de Pesquisas Energeticas e Nucleares (IPEN-CNEN/SP), Sao Paulo, SP (Brazil). Centro de Tecnologia das Radiacoes], e-mail: sborrely@ipen.br; Amaral, Ademir; Melo, Ana M.M.A. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de Energia Nuclear. Grupo de Estudos em Radioprotecao e Radioecologia (GERAR)], e-mail: amaral@ufpe.br; Silva, Luanna R.S. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de Biofisica e Radiobiologia. Lab. de Radiobiologia], e-mail: amdemelo@hotmail.com, e-mail: luannaribeiro_lua@hotmail.com

    2009-07-01

    Ionizing radiations are known as mutagenic agents, causing lethality and infertility. This characteristic has motivated its application on animal biological control. In this context, the freshwater snail Biomphalaria glabrata can be considered an excellent experimental model to study effects of ionizing radiations on lethality and reproduction. This work was designed to evaluate effects of {sup 60}Co gamma radiation at high dose rate (10.04 kGy/h) on B. glabrata. For this purpose, adult snails were selected and exposed to doses ranging from 20 to 100 Gy, with 10 Gy intervals; one group was kept as control. There was not effect of dose rate in the lethality of gamma radiation; the value of 64,3 Gy of LD{sub 50} obtained in our study was similar to that obtained by other authors with low dose rates. Nevertheless, our data suggest that there was a dose rate effect in the reproduction. On all dose levels, radiation improved the production of embryos for all exposed individuals. However, viability indexes were below 6% and, even 65 days after irradiation, fertility was not recovered. These results are not in agreement with other studies using low dose rates. Lethality was obtained in all groups irradiated, and the highest doses presented percentiles of dead animals above 50%. The results demonstrated that doses of 20 and 30 Gy were ideal for population control of B. glabrata. Further studies are needed; nevertheless, this research evidenced great potential of high dose rate gamma radiation on B. glabrata reproductive control. (author)

  10. Effects of high dose rate gamma radiation on survival and reproduction of Biomphalaria glabrata

    International Nuclear Information System (INIS)

    Cantinha, Rebeca S.; Nakano, Eliana; Silva, Luanna R.S.

    2009-01-01

    Ionizing radiations are known as mutagenic agents, causing lethality and infertility. This characteristic has motivated its application on animal biological control. In this context, the freshwater snail Biomphalaria glabrata can be considered an excellent experimental model to study effects of ionizing radiations on lethality and reproduction. This work was designed to evaluate effects of 60 Co gamma radiation at high dose rate (10.04 kGy/h) on B. glabrata. For this purpose, adult snails were selected and exposed to doses ranging from 20 to 100 Gy, with 10 Gy intervals; one group was kept as control. There was not effect of dose rate in the lethality of gamma radiation; the value of 64,3 Gy of LD 50 obtained in our study was similar to that obtained by other authors with low dose rates. Nevertheless, our data suggest that there was a dose rate effect in the reproduction. On all dose levels, radiation improved the production of embryos for all exposed individuals. However, viability indexes were below 6% and, even 65 days after irradiation, fertility was not recovered. These results are not in agreement with other studies using low dose rates. Lethality was obtained in all groups irradiated, and the highest doses presented percentiles of dead animals above 50%. The results demonstrated that doses of 20 and 30 Gy were ideal for population control of B. glabrata. Further studies are needed; nevertheless, this research evidenced great potential of high dose rate gamma radiation on B. glabrata reproductive control. (author)

  11. Enhanced Efflux Pump Activity in Old Candida glabrata Cells.

    Science.gov (United States)

    Bhattacharya, Somanon; Fries, Bettina C

    2018-03-01

    We investigated the effect of replicative aging on antifungal resistance in Candida glabrata Our studies demonstrate significantly increased transcription of ABC transporters and efflux pump activity in old versus young C. glabrata cells of a fluconazole-sensitive and -resistant strain. In addition, higher tolerance to killing by micafungin and amphotericin B was noted and is associated with higher transcription of glucan synthase gene FKS1 and lower ergosterol content in older cells. Copyright © 2018 American Society for Microbiology.

  12. Ultrastructural studies of Biomphalaria glabrata (Say, 1818) embryo

    International Nuclear Information System (INIS)

    Kikuchi, O.K.; Okazaki, K.; Kawano, T.; Ribeiro, A.A.G.F.C.

    1988-09-01

    Ultrastructural studies of Biomphalaria glabrata embryos (MOllusca: Gastropoda), and important snail vector of schistosomiasis has not been explored. In the present work it was evaluated a suitable electron microscopical technique for embryos processing. Promising results was obtained with double fixation in 1% glutaraldehyde plus 1% osmium tetroxide in 0.05 M cacodylate buffer (pH 7.4), preliminary staining overnight in 1% uranyl acetate and embedding in EPON or Polylite under vacuum. It was used embryos at young trochophore stage wich is characterized by active organogenesis. Some ultrastructural aspects of B. glabrata embryos cells are presented. (author) [pt

  13. New insights into the amphibious life of Biomphalaria glabrata and susceptibility of its egg masses to fungal infection.

    Science.gov (United States)

    Duarte, Glennyha F; Rodrigues, Juscelino; Fernandes, Éverton K K; Humber, Richard A; Luz, Christian

    2015-02-01

    The air-breathing snail Biomphalaria glabrata proliferates in stagnant freshwater, and nothing is known about the survival of eggs in intermittently (rather than perpetually) wet habitats. In the present study their egg masses matured, and juveniles subsequently eclosed and were mobile in a stable water film of transitory habitats simulated by two different simple test devices described here. The viability of eggs maintained in an unstable film however, was diminished. The maturation of egg masses in a water film or in water was significantly prevented by the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae. The efficiency depended on the fungal propagule and test environment. Hyphal bodies were more effective against egg masses than conidia. This appears to be a first report of activity of either entomopathogen against a mollusc. Both devices offer accurate and reproducible conditions to test both biological questions and the effects of substances or pathogens against B. glabrata egg masses in water films. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Dissection of Ire1 functions reveals stress response mechanisms uniquely evolved in Candida glabrata.

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    Taiga Miyazaki

    2013-01-01

    Full Text Available Proper protein folding in the endoplasmic reticulum (ER is vital in all eukaryotes. When misfolded proteins accumulate in the ER lumen, the transmembrane kinase/endoribonuclease Ire1 initiates splicing of HAC1 mRNA to generate the bZIP transcription factor Hac1, which subsequently activates its target genes to increase the protein-folding capacity of the ER. This cellular machinery, called the unfolded protein response (UPR, is believed to be an evolutionarily conserved mechanism in eukaryotes. In this study, we comprehensively characterized mutant phenotypes of IRE1 and other related genes in the human fungal pathogen Candida glabrata. Unexpectedly, Ire1 was required for the ER stress response independently of Hac1 in this fungus. C. glabrata Ire1 did not cleave mRNAs encoding Hac1 and other bZIP transcription factors identified in the C. glabrata genome. Microarray analysis revealed that the transcriptional response to ER stress is not mediated by Ire1, but instead is dependent largely on calcineurin signaling and partially on the Slt2 MAPK pathway. The loss of Ire1 alone did not confer increased antifungal susceptibility in C. glabrata contrary to UPR-defective mutants in other fungi. Taken together, our results suggest that the canonical Ire1-Hac1 UPR is not conserved in C. glabrata. It is known in metazoans that active Ire1 nonspecifically cleaves and degrades a subset of ER-localized mRNAs to reduce the ER load. Intriguingly, this cellular response could occur in an Ire1 nuclease-dependent fashion in C. glabrata. We also uncovered the attenuated virulence of the C. glabrata Δire1 mutant in a mouse model of disseminated candidiasis. This study has unveiled the unique evolution of ER stress response mechanisms in C. glabrata.

  15. Candida (Torulopsis glabrata) liver abscesses eight years after orthotopic liver transplantation.

    Science.gov (United States)

    Annunziata, G M; Blackstone, M; Hart, J; Piper, J; Baker, A L

    1997-04-01

    The authors report the case of a 48-year-old man in whom candida (Torulopsis glabrata) liver abscesses developed 8 years after liver transplantation. After a week of fever, computed tomography and Doppler ultrasonography showed several fluid-filled loculations in the left lobe of the liver and hepatic arterial stenosis. Aspirates from the abscesses contained T. glabrata organisms. This complication probably developed because hepatic arterial stenosis resulted in bile infarcts (bilomas), which were contaminated via the biliary tract with candida from the biliary-enteric anastomosis. Catheter drainage and administration of amphotericin B for 10 weeks permitted successful retransplantation. T. glabrata liver abscesses, a life threatening complication that can occur long after liver transplantation, can be successfully managed by aggressive medical treatment followed by retransplantation.

  16. Predictive Virtual Infection Modeling of Fungal Immune Evasion in Human Whole Blood

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    Maria T. E. Prauße

    2018-03-01

    Full Text Available Bloodstream infections by the human-pathogenic fungi Candida albicans and Candida glabrata increasingly occur in hospitalized patients and are associated with high mortality rates. The early immune response against these fungi in human blood comprises a concerted action of humoral and cellular components of the innate immune system. Upon entering the blood, the majority of fungal cells will be eliminated by innate immune cells, i.e., neutrophils and monocytes. However, recent studies identified a population of fungal cells that can evade the immune response and thereby may disseminate and cause organ dissemination, which is frequently observed during candidemia. In this study, we investigate the so far unresolved mechanism of fungal immune evasion in human whole blood by testing hypotheses with the help of mathematical modeling. We use a previously established state-based virtual infection model for whole-blood infection with C. albicans to quantify the immune response and identified the fungal immune-evasion mechanism. While this process was assumed to be spontaneous in the previous model, we now hypothesize that the immune-evasion process is mediated by host factors and incorporate such a mechanism in the model. In particular, we propose, based on previous studies that the fungal immune-evasion mechanism could possibly arise through modification of the fungal surface by as of yet unknown proteins that are assumed to be secreted by activated neutrophils. To validate or reject any of the immune-evasion mechanisms, we compared the simulation of both immune-evasion models for different infection scenarios, i.e., infection of whole blood with either C. albicans or C. glabrata under non-neutropenic and neutropenic conditions. We found that under non-neutropenic conditions, both immune-evasion models fit the experimental data from whole-blood infection with C. albicans and C. glabrata. However, differences between the immune-evasion models could be

  17. Predictive Virtual Infection Modeling of Fungal Immune Evasion in Human Whole Blood.

    Science.gov (United States)

    Prauße, Maria T E; Lehnert, Teresa; Timme, Sandra; Hünniger, Kerstin; Leonhardt, Ines; Kurzai, Oliver; Figge, Marc Thilo

    2018-01-01

    Bloodstream infections by the human-pathogenic fungi Candida albicans and Candida glabrata increasingly occur in hospitalized patients and are associated with high mortality rates. The early immune response against these fungi in human blood comprises a concerted action of humoral and cellular components of the innate immune system. Upon entering the blood, the majority of fungal cells will be eliminated by innate immune cells, i.e., neutrophils and monocytes. However, recent studies identified a population of fungal cells that can evade the immune response and thereby may disseminate and cause organ dissemination, which is frequently observed during candidemia. In this study, we investigate the so far unresolved mechanism of fungal immune evasion in human whole blood by testing hypotheses with the help of mathematical modeling. We use a previously established state-based virtual infection model for whole-blood infection with C. albicans to quantify the immune response and identified the fungal immune-evasion mechanism. While this process was assumed to be spontaneous in the previous model, we now hypothesize that the immune-evasion process is mediated by host factors and incorporate such a mechanism in the model. In particular, we propose, based on previous studies that the fungal immune-evasion mechanism could possibly arise through modification of the fungal surface by as of yet unknown proteins that are assumed to be secreted by activated neutrophils. To validate or reject any of the immune-evasion mechanisms, we compared the simulation of both immune-evasion models for different infection scenarios, i.e., infection of whole blood with either C. albicans or C. glabrata under non-neutropenic and neutropenic conditions. We found that under non-neutropenic conditions, both immune-evasion models fit the experimental data from whole-blood infection with C. albicans and C. glabrata . However, differences between the immune-evasion models could be observed for the

  18. Fluconazole Resistance Associated with Drug Efflux and Increased Transcription of a Drug Transporter Gene, PDH1, in Candida glabrata

    Science.gov (United States)

    Miyazaki, Haruko; Miyazaki, Yoshitsugu; Geber, Antonia; Parkinson, Tanya; Hitchcock, Christopher; Falconer, Derek J.; Ward, Douglas J.; Marsden, Katherine; Bennett, John E.

    1998-01-01

    Sequential Candida glabrata isolates were obtained from the mouth of a patient infected with human immunodeficiency virus type 1 who was receiving high doses of fluconazole for oropharyngeal thrush. Fluconazole-susceptible colonies were replaced by resistant colonies that exhibited both increased fluconazole efflux and increased transcripts of a gene which codes for a protein with 72.5% identity to Pdr5p, an ABC multidrug transporter in Saccharomyces cerevisiae. The deduced protein had a molecular mass of 175 kDa and was composed of two homologous halves, each with six putative transmembrane domains and highly conserved sequences of ATP-binding domains. When the earliest and most azole-susceptible isolate of C. glabrata from this patient was exposed to fluconazole, increased transcripts of the PDR5 homolog appeared, linking azole exposure to regulation of this gene. PMID:9661006

  19. Differentiation of Candida albicans, Candida glabrata, and Candida krusei by FT-IR and chemometrics by CHROMagar™ Candida.

    Science.gov (United States)

    Wohlmeister, Denise; Vianna, Débora Renz Barreto; Helfer, Virginia Etges; Calil, Luciane Noal; Buffon, Andréia; Fuentefria, Alexandre Meneghello; Corbellini, Valeriano Antonio; Pilger, Diogo André

    2017-10-01

    Pathogenic Candida species are detected in clinical infections. CHROMagar™ is a phenotypical method used to identify Candida species, although it has limitations, which indicates the need for more sensitive and specific techniques. Infrared Spectroscopy (FT-IR) is an analytical vibrational technique used to identify patterns of metabolic fingerprint of biological matrixes, particularly whole microbial cell systems as Candida sp. in association of classificatory chemometrics algorithms. On the other hand, Soft Independent Modeling by Class Analogy (SIMCA) is one of the typical algorithms still little employed in microbiological classification. This study demonstrates the applicability of the FT-IR-technique by specular reflectance associated with SIMCA to discriminate Candida species isolated from vaginal discharges and grown on CHROMagar™. The differences in spectra of C. albicans, C. glabrata and C. krusei were suitable for use in the discrimination of these species, which was observed by PCA. Then, a SIMCA model was constructed with standard samples of three species and using the spectral region of 1792-1561cm -1 . All samples (n=48) were properly classified based on the chromogenic method using CHROMagar™ Candida. In total, 93.4% (n=45) of the samples were correctly and unambiguously classified (Class I). Two samples of C. albicans were classified correctly, though these could have been C. glabrata (Class II). Also, one C. glabrata sample could have been classified as C. krusei (Class II). Concerning these three samples, one triplicate of each was included in Class II and two in Class I. Therefore, FT-IR associated with SIMCA can be used to identify samples of C. albicans, C. glabrata, and C. krusei grown in CHROMagar™ Candida aiming to improve clinical applications of this technique. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. [Comparison of microdilution and disk diffusion methods for the detection of fluconazole and voriconazole susceptibility against clinical Candida glabrata isolates and determination of changing susceptibility with new CLSI breakpoints].

    Science.gov (United States)

    Hazırolan, Gülşen; Sarıbaş, Zeynep; Arıkan Akdağlı, Sevtap

    2016-07-01

    Candida albicans is the most frequently isolated species as the causative agent of Candida infections. However, in recent years, the isolation rate of non-albicans Candida species have increased. In many centers, Candida glabrata is one of the commonly isolated non-albicans species of C.glabrata infections which are difficult-to-treat due to decreased susceptibility to fluconazole and cross-resistance to other azoles. The aims of this study were to determine the in vitro susceptibility profiles of clinical C.glabrata isolates against fluconazole and voriconazole by microdilution and disk diffusion methods and to evaluate the results with both the previous (CLSI) and current species-specific CLSI (Clinical and Laboratory Standards Institute) clinical breakpoints. A total of 70 C.glabrata strains isolated from clinical samples were included in the study. The identification of the isolates was performed by morphologic examination on cornmeal Tween 80 agar and assimilation profiles obtained by using ID32C (BioMérieux, France). Broth microdilution and disk diffusion methods were performed according to CLSI M27-A3 and CLSI M44-A2 documents, respectively. The results were evaluated according to CLSI M27-A3 and M44-A2 documents and new vs. species-specific CLSI breakpoints. By using both previous and new CLSI breakpoints, broth microdilution test results showed that voriconazole has greater in vitro activity than fluconazole against C.glabrata isolates. For the two drugs tested, very major error was not observed with disk diffusion method when microdilution method was considered as the reference method. Since "susceptible" category no more exists for fluconazole vs. C.glabrata, the isolates that were interpreted as susceptible by previous breakpoints were evaluated as susceptible-dose dependent by current CLSI breakpoints. Since species-specific breakpoints remain yet undetermined for voriconazole, comparative analysis was not possible for this agent. The results obtained

  1. Differences in the gene expression profiles of haemocytes from schistosome-susceptible and -resistant biomphalaria glabrata exposed to Schistosoma mansoni excretory-secretory products.

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    Zahida Zahoor

    Full Text Available During its life cycle, the helminth parasite Schistosoma mansoni uses the freshwater snail Biomphalaria glabrata as an intermediate host to reproduce asexually generating cercariae for infection of the human definitive host. Following invasion of the snail, the parasite develops from a miracidium to a mother sporocyst and releases excretory-secretory products (ESPs that likely influence the outcome of host infection. To better understand molecular interactions between these ESPs and the host snail defence system, we determined gene expression profiles of haemocytes from S. mansoni-resistant or -susceptible strains of B. glabrata exposed in vitro to S. mansoni ESPs (20 μg/ml for 1 h, using a 5K B. glabrata cDNA microarray. Ninety-eight genes were found differentially expressed between haemocytes from the two snail strains, 57 resistant specific and 41 susceptible specific, 60 of which had no known homologue in GenBank. Known differentially expressed resistant-snail genes included the nuclear factor kappa B subunit Relish, elongation factor 1α, 40S ribosomal protein S9, and matrilin; known susceptible-snail specific genes included cathepsins D and L, and theromacin. Comparative analysis with other gene expression studies revealed 38 of the 98 identified genes to be uniquely differentially expressed in haemocytes in the presence of ESPs, thus identifying for the first time schistosome ESPs as important molecules that influence global snail host-defence cell gene expression profiles. Such immunomodulation may benefit the schistosome, enabling its survival and successful development in the snail host.

  2. Candida species isolated from the vaginal mucosa of HIV-infected women in Salvador, Bahia, Brazil

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    Paula Matos Oliveira

    Full Text Available BACKGROUND: Vulvovaginal candidiasis (VVC is the second most common vaginal infection. HIV-infection is a risk factor for this infection. OBJECTIVE: To determine the frequency of VVC and to describe the main Candida species isolated and their susceptibility to antifungal drugs in HIV-infected patients, compared to HIV-uninfected women in Salvador, Brazil. METHODS: Cross-sectional study including a group of 64 HIV-infected women and 76 uninfected women, followed up at the AIDS reference center and at the Gynecological Clinic of Escola Bahiana de Medicina e Saúde Pública (Salvador, Bahia, Brazil. RESULTS: Frequency of Candida spp. was higher in HIV-infected women (29.7% than in HIV-uninfected controls (14.5% (p = 0.02. The odds ratio value for vulvovaginal candidiasis in HIV-infected patients was 2.6 (95% CI: 1.07 - 6.32 p = 0.03. Candida albicans was the most commonly isolated species in both HIV-infected (52.3% and uninfected women (85.7%, followed by C. parapsolis in 17.6% and 14.3%, respectively. In HIV-infected women, C. glabrata, C. parapsilosis, and a coinfection of C. albicans and C. glabrata were also identified. There was no significant difference between Candida species isolated from the vaginal mucosa of women with VVC and colonization of the vaginal mucosa of HIV-infected and HIV-uninfected women. One C. glabrata isolate from an HIV-infected patient was resistant to fluconazole and other two isolates exhibited a dose-dependent susceptibility. CONCLUSION: Our results confirm a higher frequency of Candida spp. isolated from the vaginal mucosa of HIV-infected women and a broader spectrum of species involved. Only Candida glabrata isolates showed decreased susceptibility to fluconazole.

  3. Candida glabrata-pneumonia in a non-immuno-suppressed patient: Imaging by digital radiology and CT

    International Nuclear Information System (INIS)

    Bankier, A.; Fleischmann, D.; Wiesmayr, M.; Laczika, K.; Huebsch, P.

    1994-01-01

    Pneumonias caused by Candida glabrata are extremely rare and occur almost exclusively in immunocompromised patients. We report an atypical case of Candida glabrata pneumonia in a non-immunocompromised patient and describe the imaging findings on digital radiography and computed tomography. (orig.) [de

  4. Small chromosomes among Danish Candida glabrata isolates originated through different mechanisms

    DEFF Research Database (Denmark)

    Ahmad, Khadija Mohamed; Ishchuk, Olena P.; Hellborg, Linda

    2013-01-01

    chromosomes, which were smaller than 0.5 Mb. Regarding the year, patient and hospital, these C. glabrata strains had independent origin and the analyzed small chromosomes were structurally not related to each other (i.e. they contained different sets of genes). We suggest that at least two mechanisms could...... exhibited mitotic instability, while the second type, which contained the corresponding genes in only one copy in the genome, was mitotically stable. Apparently, in patients C. glabrata chromosomes are frequently reshuffled resulting in new genetic configurations, including appearance of small chromosomes...

  5. Use of Biomphalaria glabrata hemocytes as a biomarker of exposure to ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Hianna A.M.F.; Lima, Maira V.; Sa, Jose L.F.; Siqueira, Williams N.; Luna Filho, Ricardo L.C.; Melo, Larissa S.A.; Morais, Vinicius H.T.; Melo, Ana M.M.A., E-mail: hiannaamfs@gmail.com, E-mail: mairavasconceloslima@gmail.com, E-mail: luismuma6@gmail.com, E-mail: williams.wns@gmail.com, E-mail: ricardolclf@hotmail.com, E-mail: larissamelo.pe@gmail.com, E-mail: viniciushtmorais@hotmail.com, E-mail: amdemelo@hotmail.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Biofisica e Radiobiologia

    2017-11-01

    The increase of the applications of ionizing radiation in several areas and sectors of modern society has given rise to a greater probability of occurrence of accidents. These accidental occurrences have revealed the need for methods that provide quantitative data on the radiation doses absorbed by biological systems. The mollusk Biomphalaria glabrata presents as a good bioindicator in several works referenced in the literature. In this way, the objective of this work was to evaluate the morphological and quantitative alterations of hemocytes of the Biomphalaria glabrata exposed to ionizing radiation. For the experiments, adult mollusks of the species B. glabrata pigmented were used. The selected mollusks were divided into six groups: five submitted to doses of 10, 20, 30, 40, 50 Gy of {sup 60}Co gamma radiation and the control group. After 48 h, the slides were prepared and then read in a microscope. Quantitative analysis showed a decrease in the total number of hemocytes after irradiation. In the cell classification, a higher number of hyalinocytes were observed in relation to the number of granulocytes, except for the animals exposed to a dose of 30 Gy. The presence of micronuclei and binucleations were observed at all doses used. Apoptosis was observed at doses starting at 30 Gy. Therefore, it is possible to conclude that the morphological and quantitative analysis of B. glabrata hemocytes provided significant data for the identification of biological damage caused by ionizing radiation, allowing the beginning of standardization of the morphological alteration counting technique in B. glabrata hemocytes as An environmental biomarker for the action of physical agents. (author)

  6. Use of Biomphalaria glabrata hemocytes as a biomarker of exposure to ionizing radiation

    International Nuclear Information System (INIS)

    Silva, Hianna A.M.F.; Lima, Maira V.; Sa, Jose L.F.; Siqueira, Williams N.; Luna Filho, Ricardo L.C.; Melo, Larissa S.A.; Morais, Vinicius H.T.; Melo, Ana M.M.A.

    2017-01-01

    The increase of the applications of ionizing radiation in several areas and sectors of modern society has given rise to a greater probability of occurrence of accidents. These accidental occurrences have revealed the need for methods that provide quantitative data on the radiation doses absorbed by biological systems. The mollusk Biomphalaria glabrata presents as a good bioindicator in several works referenced in the literature. In this way, the objective of this work was to evaluate the morphological and quantitative alterations of hemocytes of the Biomphalaria glabrata exposed to ionizing radiation. For the experiments, adult mollusks of the species B. glabrata pigmented were used. The selected mollusks were divided into six groups: five submitted to doses of 10, 20, 30, 40, 50 Gy of "6"0Co gamma radiation and the control group. After 48 h, the slides were prepared and then read in a microscope. Quantitative analysis showed a decrease in the total number of hemocytes after irradiation. In the cell classification, a higher number of hyalinocytes were observed in relation to the number of granulocytes, except for the animals exposed to a dose of 30 Gy. The presence of micronuclei and binucleations were observed at all doses used. Apoptosis was observed at doses starting at 30 Gy. Therefore, it is possible to conclude that the morphological and quantitative analysis of B. glabrata hemocytes provided significant data for the identification of biological damage caused by ionizing radiation, allowing the beginning of standardization of the morphological alteration counting technique in B. glabrata hemocytes as An environmental biomarker for the action of physical agents. (author)

  7. An unexpected response of torulopsis glabrata fusion products to x-irradiation

    International Nuclear Information System (INIS)

    Galeotti, C.L.; Sriprakash, K.S.; Batum, C.M.; Clark-Walker, G.D.

    1981-01-01

    Intra-species fusion products of Saccharomyces cerevisiae, Saccharomyces unisporus and Torulopsis glabrata have been isolated following polyethylene glycol-induced fusion of protoplasts and selection for prototrophic colonies. Staining with lomofungin showed that all fusion products were uninucleate. Measurement of DNA content mostly gave values between haploid and diploid levels indicating that the majority of fusion products were aneuploid. Nevertheless fusion products of S. cerevisiae and S. unisporus were, as expected, more resistant to X-irradiation than their haploid parents. By contrast, the X-ray doze-response curve of all T. glabrata fusion products was indistinguishable from their progenitors despite the fact that mitotic segregants could be recovered amongst the survivors to X-rays. A possible explanation for the behaviour towards X-rays of T. glabrata fusion products is that this species lacks a DNA repair pathway involving recombination between homologous chromosomes. We conclude from this study that the shape of the X-ray dose-response curve should not be taken to indicate the ploidy of new yeast isolates without supporting data. (orig.)

  8. Genotyping of the MTL loci and susceptibility to two antifungal agents of Candida glabrata clinical isolates

    Directory of Open Access Journals (Sweden)

    María Teresa Lavaniegos-Sobrino

    2009-08-01

    Full Text Available The opportunistic fungal pathogen Candida glabrata is the second most common isolate from bloodstream infections worldwide and is naturally less susceptible to the antifungal drug fluconazole than other Candida species. C. glabrata is a haploid yeast that contains three mating-type like loci (MTL, although no sexual cycle has been described. Strains containing both types of mating information at the MTL1 locus are found in clinical isolates, but it is thought that strains containing type a information are more common. Here we investigated if a particular combination of mating type information at each MTLlocus is more prevalent in clinical isolates from hospitalized patients in Mexico and if there is a correlation between mating information and resistance to fluconazole and 5-fluorocytosine. We found that while both types of information at MTL1 are equally represented in a collection of 64 clinical isolates, the vast majority of isolates contain a-type information at MTL2 and α-type at MTL3. We also found no correlation of the particular combination of mating type information at the three MTL loci and resistance to fluconazole.

  9. Reversal of fluconazole resistance induced by a synergistic effect with Acca sellowiana in Candida glabrata strains.

    Science.gov (United States)

    R M Machado, Gabriella da; Pippi, Bruna; Dalla Lana, Daiane Flores; Amaral, Ana Paula S; Teixeira, Mário Lettieri; Souza, Kellen C B de; Fuentefria, Alexandre M

    2016-11-01

    The increased incidence of non-albicans Candida (NAC) resistant to fluconazole (FLZ) makes it necessary to use new therapeutic alternatives. Acca sellowiana (O.berg) Burret (Myrtaceae) is a guava with several proven biological activities. The interaction with fluconazole can be a feasible alternative to overcome this resistance. This study evaluates the in vitro antifungal activity of fractions obtained from the lyophilized aqueous extract of the leaves of A. sellowiana against resistant strains of NAC. The antifungal activity of the fractions was evaluated at 500 μg/mL by microdilution method. Checkerboard assay was performed to determine the effect of the combination of the F2 fraction and antifungal at concentrations: MIC/4, MIC/2, MIC, MIC × 2 and MIC × 4. Candida glabrata showed the lowest MIC values (500-3.90 μg/mL) and the F2 active fraction was the most effective. The association of F2 with FLZ showed a strong synergistic effect (FICI ≤ 0.5) against 100% of C. glabrata resistant isolates. Moreover, the F2 active fraction has demonstrated that probably acts in the cell wall of these yeasts. There was no observed acute dermal toxicity of lyophilized aqueous extract of leaves of A. sellowiana on pig ear skin cells. The interaction between substances present in the F2 active fraction is possibly responsible for the antifungal activity presented by this fraction. This study is unprecedented and suggests that the combination of F2 active fraction and FLZ might be used as an alternative treatment for mucocutaneus infections caused by C. glabrata resistant.

  10. Quantitative Simulations Predict Treatment Strategies Against Fungal Infections in Virtual Neutropenic Patients.

    Science.gov (United States)

    Timme, Sandra; Lehnert, Teresa; Prauße, Maria T E; Hünniger, Kerstin; Leonhardt, Ines; Kurzai, Oliver; Figge, Marc Thilo

    2018-01-01

    The condition of neutropenia, i.e., a reduced absolute neutrophil count in blood, constitutes a major risk factor for severe infections in the affected patients. Candida albicans and Candida glabrata are opportunistic pathogens and the most prevalent fungal species in the human microbiota. In immunocompromised patients, they can become pathogenic and cause infections with high mortality rates. In this study, we use a previously established approach that combines experiments and computational models to investigate the innate immune response during blood stream infections with the two fungal pathogens C. albicans and C. glabrata . First, we determine immune-reaction rates and migration parameters under healthy conditions. Based on these findings, we simulate virtual patients and investigate the impact of neutropenic conditions on the infection outcome with the respective pathogen. Furthermore, we perform in silico treatments of these virtual patients by simulating a medical treatment that enhances neutrophil activity in terms of phagocytosis and migration. We quantify the infection outcome by comparing the response to the two fungal pathogens relative to non-neutropenic individuals. The analysis reveals that these fungal infections in neutropenic patients can be successfully cleared by cytokine treatment of the remaining neutrophils; and that this treatment is more effective for C. glabrata than for C. albicans .

  11. Development of a Candida glabrata dominant nutritional transformation marker utilizing the Aspergillus nidulans acetamidase gene (amdS).

    Science.gov (United States)

    Fu, Jianmin; Blaylock, Morganne; Wickes, Cameron F; Welte, William; Mehrtash, Adrian; Wiederhold, Nathan; Wickes, Brian L

    2016-05-01

    The gene encoding Aspergillus nidulans acetamidase (amdS) was placed under control of Candida albicans ACT1 promoter and terminator sequences and then cloned into a plasmid containing C. glabrata ARS10,CEN8 or ARS10+CEN8 sequences. All plasmids transformed C. glabrata wild-type cells to acetamide+, with the ARS-only containing plasmid transforming cells at the highest frequencies (>1.0 × 10(4) transformants μg(-1)). Plasmids were rapidly lost under non-selective conditions with the frequency dependent on chromosomal element, thus recycling the acetamide- phenotype. The amdS plasmid was used to transform a set of clinical isolates resistant to a variety of antifungal drugs. All strains were successfully transformed to the acetamide+ phenotype at high frequency, confirming that this plasmid construct could be used as a simple dominant marker on virtually any strain. Gap repair experiments demonstrated that just as in Saccharomyces cerevisiae, gap repair functions efficiently inC. glabrata, suggesting that C. glabrata has numerous similarities toS. cerevisiae with regard to ease of molecular manipulation. The amdS system is inexpensive and efficient, and combined with existing C. glabrata plasmid elements, confers a high transformation frequency for C. glabrata with a phenotype that can be easily recycled. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  12. Probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 exhibit strong antifungal effects against vulvovaginal candidiasis-causing Candida glabrata isolates.

    Science.gov (United States)

    Chew, S Y; Cheah, Y K; Seow, H F; Sandai, D; Than, L T L

    2015-05-01

    This study investigates the antagonistic effects of the probiotic strains Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 against vulvovaginal candidiasis (VVC)-causing Candida glabrata. Growth inhibitory activities of Lact. rhamnosus GR-1 and Lact. reuteri RC-14 strains against C. glabrata were demonstrated using a spot overlay assay and a plate-based microtitre assay. In addition, these probiotic lactobacilli strains also exhibited potent candidacidal activity against C. glabrata, as demonstrated by a LIVE/DEAD yeast viability assay performed using confocal laser scanning microscopy. The metabolic activities of all C. glabrata strains were completely shut down in response to the challenges by the probiotic lactobacilli strains. In addition, both probiotic lactobacilli strains exhibited strong autoaggregation and coaggregation phenotypes in the presence of C. glabrata, which indicate that these lactobacilli strains may exert their probiotic effects through the formation of aggregates and, thus the consequent prevention of colonization by C. glabrata. Probiotic Lact. rhamnosus GR-1 and Lact. reuteri RC-14 strains exhibited potent antagonistic activities against all of the tested C. glabrata strains. These lactobacilli exhibited antifungal effects, including those attributed to their aggregation abilities, and their presence caused the cessation of growth and eventual cell death of C. glabrata. This is the first study to report on the antagonistic effects of these probiotic lactobacilli strains against the non-Candida albicans Candida (NCAC) species C. glabrata. © 2015 The Authors published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.

  13. Consumption of Biomphalaria glabrata egg masses and juveniles by the ampullariid snails Pila ovata, Lanistes carinatus and Marisa cornuarietis.

    Science.gov (United States)

    Hofkin, B V; Stryker, G A; Koech, D K; Loker, E S

    1991-04-01

    Lanistes carinatus and Pila ovata from Kenya and Marisa cornuarietis from the Dominican Republic were examined for their ability to consume egg masses and juveniles of Biomphalaria glabrata (M line strain) under laboratory conditions. Adults of all three ampullariid species consumed all egg masses presented to them over a five day period of observation. Juvenile P. ovata consumed significantly more egg masses than juveniles of M. cornuarietis or L. carinatus. Both adult and juvenile ampullariids generally ate more egg masses when maintained under a temperature regime (25-32 degrees C) approximating conditions in coastal Kenya than at 13-25 degrees C which approximates the temperature regime near Nairobi, Kenya. Egg masses attached to floating 'refugia' were not attacked, apparently reflecting the difficulty experienced by ampullariids in reaching free-floating objects. Adults of each ampullariid species ate approximately 25% of all lariids in reaching free-floating objects. Adults of each ampullariid species ate approximately 25% of all 1.5 +/- 0.5 mm B. glabrata juvenile snails presented to them. P. ovata adults consumed significantly more 3.0 +/- 0.5 mm juveniles than adults of the other two species. B. glabrata egg masses and juveniles were consumed even though lettuce was continually present in experimental aquaria. Implications of these results for biological control studies in East Africa are discussed.

  14. Identification of Candida glabrata genes involved in pH modulation and modification of the phagosomal environment in macrophages.

    Directory of Open Access Journals (Sweden)

    Lydia Kasper

    Full Text Available Candida glabrata currently ranks as the second most frequent cause of invasive candidiasis. Our previous work has shown that C. glabrata is adapted to intracellular survival in macrophages and replicates within non-acidified late endosomal-stage phagosomes. In contrast, heat killed yeasts are found in acidified matured phagosomes. In the present study, we aimed at elucidating the processes leading to inhibition of phagosome acidification and maturation. We show that phagosomes containing viable C. glabrata cells do not fuse with pre-labeled lysosomes and possess low phagosomal hydrolase activity. Inhibition of acidification occurs independent of macrophage type (human/murine, differentiation (M1-/M2-type or activation status (vitamin D3 stimulation. We observed no differential activation of macrophage MAPK or NFκB signaling cascades downstream of pattern recognition receptors after internalization of viable compared to heat killed yeasts, but Syk activation decayed faster in macrophages containing viable yeasts. Thus, delivery of viable yeasts to non-matured phagosomes is likely not triggered by initial recognition events via MAPK or NFκB signaling, but Syk activation may be involved. Although V-ATPase is abundant in C. glabrata phagosomes, the influence of this proton pump on intracellular survival is low since blocking V-ATPase activity with bafilomycin A1 has no influence on fungal viability. Active pH modulation is one possible fungal strategy to change phagosome pH. In fact, C. glabrata is able to alkalinize its extracellular environment, when growing on amino acids as the sole carbon source in vitro. By screening a C. glabrata mutant library we identified genes important for environmental alkalinization that were further tested for their impact on phagosome pH. We found that the lack of fungal mannosyltransferases resulted in severely reduced alkalinization in vitro and in the delivery of C. glabrata to acidified phagosomes. Therefore

  15. Prevalence of Candida co-infection in patients with pulmonary tuberculosis.

    Science.gov (United States)

    Kali, Arunava; Charles, Mv Pravin; Noyal, Mariya Joseph; Sivaraman, Umadevi; Kumar, Shailesh; Easow, Joshy M

    2013-01-01

    Candida species are emerging as a potentially pathogenic fungus in patients with broncho-pulmonary diseases. The synergistic growth promoting association of Candida and Mycobacterium tuberculosis has raised increased concern for studying the various Candida spp . and its significance in pulmonary tuberculosis patients during current years. This study was undertaken with the objective of discovering the prevalence of co-infection caused by different Candida species in patients with pulmonary tuberculosis. A total of 75 patients with pulmonary tuberculosis diagnosed by sputum Ziehl-Neelsen staining were included in the study. Candida co-infection was confirmed using the Kahanpaa et al. criteria. Candida species were identified using gram stain morphology, germ tube formation, morphology on cornmeal agar with Tween-80, sugar fermentation tests and HiCrome Candida Agar. Candida co-infection was observed in 30 (40%) of patients with pulmonary tuberculosis. Candida albicans was the most common isolate observed in 50% of the patients with co-infection, followed by C. tropicalis (20%) and C. glabrata (20%). Candida co-infection was found in 62.5% of female patients, while it was observed in only 29.4% of the male patients (P value 0.0133). Mean ± SD age of the patients with C. glabrata infection was 65.83 ± 3.19, while the mean ± SD age of the patients with other Candida infections was 43.25 ± 20.44 (P value 0.0138). Many patients with pulmonary tuberculosis have co-infection with Candida spp. The prevalence of non-albicans Candida species is increasing and may be associated with inadequate response to anti-tubercular drugs. C. glabrata infection has a strong association with old age.

  16. A network of paralogous stress response transcription factors in the human pathogen Candida glabrata.

    Directory of Open Access Journals (Sweden)

    Jawad eMerhej

    2016-05-01

    Full Text Available The yeast Candida glabrata has become the second cause of systemic candidemia in humans. However, relatively few genome-wide studies have been conducted in this organism and our knowledge of its transcriptional regulatory network is quite limited. In the present work, we combined genome-wide chromatin immunoprecipitation (ChIP-seq, transcriptome analyses and DNA binding motif predictions to describe the regulatory interactions of the seven Yap (Yeast AP1 transcription factors of C. glabrata. We described a transcriptional network containing 255 regulatory interactions and 309 potential target genes. We predicted with high confidence the preferred DNA binding sites for 5 of the 7 CgYaps and showed a strong conservation of the Yap DNA binding properties between S. cerevisiae and C. glabrata. We provided reliable functional annotation for 3 of the 7 Yaps and identified for Yap1 and Yap5 a core regulon which is conserved in S. cerevisiae, C. glabrata and C. albicans. We uncovered new roles for CgYap7 in the regulation of iron-sulfur cluster biogenesis, for CgYap1 in the regulation of heme biosynthesis and for CgYap5 in the repression of GRX4 in response to iron starvation. These transcription factors define an interconnected transcriptional network at the cross-roads between redox homeostasis, oxygen consumption and iron metabolism.

  17. Unexpected effects of azole transporter inhibitors on antifungal susceptibility in Candida glabrata and other pathogenic Candida species.

    Science.gov (United States)

    Nagayoshi, Yohsuke; Miyazaki, Taiga; Shimamura, Shintaro; Nakayama, Hironobu; Minematsu, Asuka; Yamauchi, Shunsuke; Takazono, Takahiro; Nakamura, Shigeki; Yanagihara, Katsunori; Kohno, Shigeru; Mukae, Hiroshi; Izumikawa, Koichi

    2017-01-01

    The pathogenic fungus Candida glabrata is often resistant to azole antifungal agents. Drug efflux through azole transporters, such as Cdr1 and Cdr2, is a key mechanism of azole resistance and these genes are under the control of the transcription factor Pdr1. Recently, the monoamine oxidase A (MAO-A) inhibitor clorgyline was shown to inhibit the azole efflux pumps, leading to increased azole susceptibility in C. glabrata. In the present study, we have evaluated the effects of clorgyline on susceptibility of C. glabrata to not only azoles, but also to micafungin and amphotericin B, using wild-type and several mutant strains. The addition of clorgyline to the culture media increased fluconazole susceptibility of a C. glabrata wild-type strain, whereas micafungin and amphotericin B susceptibilities were markedly decreased. These phenomena were also observed in other medically important Candida species, including Candida albicans, Candida parapsilosis, Candida tropicalis, and Candida krusei. Expression levels of CDR1, CDR2 and PDR1 mRNAs and an amount of Cdr1 protein in the C. glabrata wild-type strain were highly increased in response to the treatment with clorgyline. However, loss of Cdr1, Cdr2, Pdr1, and a putative clorgyline target (Fms1), which is an ortholog of human MAO-A, or overexpression of CDR1 did not affect the decreased susceptibility to micafungin and amphotericin B in the presence of clorgyline. The presence of other azole efflux pump inhibitors including milbemycin A4 oxime and carbonyl cyanide 3-chlorophenylhydrazone also decreased micafungin susceptibility in C. glabrata wild-type, Δcdr1, Δcdr2, and Δpdr1 strains. These findings suggest that azole efflux pump inhibitors increase azole susceptibility but concurrently induce decreased susceptibility to other classes of antifungals independent of azole transporter functions.

  18. Metabolic engineering of Candida glabrata for diacetyl production.

    Directory of Open Access Journals (Sweden)

    Xiang Gao

    Full Text Available In this study, Candida glabrata, an efficient pyruvate-producing strain, was metabolically engineered for the production of the food ingredient diacetyl. A diacetyl biosynthetic pathway was reconstructed based on genetic modifications and medium optimization. The former included (i channeling carbon flux into the diacetyl biosynthetic pathway by amplification of acetolactate synthase, (ii elimination of the branched pathway of α-acetolactate by deleting the ILV5 gene, and (iii restriction of diacetyl degradation by deleting the BDH gene. The resultant strain showed an almost 1∶1 co-production of α-acetolactate and diacetyl (0.95 g L(-1. Furthermore, addition of Fe3+ to the medium enhanced the conversion of α-acetolactate to diacetyl and resulted in a two-fold increase in diacetyl production (2.1 g L(-1. In addition, increased carbon flux was further channeled into diacetyl biosynthetic pathway and a titer of 4.7 g L(-1 of diacetyl was achieved by altering the vitamin level in the flask culture. Thus, this study illustrates that C. glabrata could be tailored as an attractive platform for enhanced biosynthesis of beneficial products from pyruvate by metabolic engineering strategies.

  19. Epidemiological investigation of Candida species causing bloodstream infection in paediatric small bowel transplant recipients.

    Science.gov (United States)

    Suhr, Mallory J; Gomes-Neto, João Carlos; Banjara, Nabaraj; Florescu, Diana F; Mercer, David F; Iwen, Peter C; Hallen-Adams, Heather E

    2017-06-01

    Small bowel transplantation (SBT) can be a life-saving medical procedure. However, these recipients experience high risk of bloodstream infections caused by Candida. This research aims to characterise the SBT recipient gut microbiota over time following transplantation and investigate the epidemiology of candidaemia in seven paediatric patients. Candida species from the recipients' ileum and bloodstream were identified by internal transcribed spacer sequence and distinguished to strain by multilocus sequence typing and randomly amplified polymorphic DNA. Antifungal susceptibility of bloodstream isolates was determined against nine antifungals. Twenty-two ileostomy samples harboured at least one Candida species. Fungaemia were caused by Candida parapsilosis, Candida albicans, Candida glabrata, Candida orthopsilosis and Candida pelliculosa. All but three bloodstream isolates showed susceptibility to all the antifungals tested. One C. glabrata isolate showed multidrug resistance to itraconazole, amphotericin B and posaconazole and intermediate resistance to caspofungin. Results are congruent with both endogenous (C. albicans, C. glabrata) and exogenous (C. parapsilosis) infections; results also suggest two patients were infected by the same strain of C. parapsilosis. Continuing to work towards a better understanding of sources of infection-particularly the exogenous sources-would lead to targeted prevention strategies. © 2017 Blackwell Verlag GmbH.

  20. The Effect of Molybdenum Fertilization on Arachis Glabrata Biomass ...

    African Journals Online (AJOL)

    The effect of molybdenum fertilization on biomass and the number of nodules of Arachis glabrata was assessed at the Teaching and Research Farm of the University of Dschang in 2011 at different periods of mowing. A factorial design comparing four doses of molybdenum as ammonium molybdate (0, 0.75, 1.5 and 2.25 ...

  1. Fluconazole affects the alkali-metal-cation homeostasis and susceptibility to cationic toxic compounds of Candida glabrata.

    Science.gov (United States)

    Elicharova, Hana; Sychrova, Hana

    2014-08-01

    Candida glabrata is a salt-tolerant and fluconazole (FLC)-resistant yeast species. Here, we analyse the contribution of plasma-membrane alkali-metal-cation exporters, a cation/proton antiporter and a cation ATPase to cation homeostasis and the maintenance of membrane potential (ΔΨ). Using a series of single and double mutants lacking CNH1 and/or ENA1 genes we show that the inability to export potassium and toxic alkali-metal cations leads to a slight hyperpolarization of the plasma membrane of C. glabrata cells; this hyperpolarization drives more cations into the cells and affects cation homeostasis. Surprisingly, a much higher hyperpolarization of C. glabrata plasma membrane was produced by incubating cells with subinhibitory concentrations of FLC. FLC treatment resulted in a substantially increased sensitivity of cells to various cationic drugs and toxic cations that are driven into the cell by negative-inside plasma-membrane potential. The effect of the combination of FLC plus cationic drug treatment was enhanced by the malfunction of alkali-metal-cation transporters that contribute to the regulation of membrane potential and cation homeostasis. In summary, we show that the combination of subinhibitory concentrations of FLC and cationic drugs strongly affects the growth of C. glabrata cells. © 2014 The Authors.

  2. Occurrence and characterization of Candida nivariensis from a culture collection of Candida glabrata clinical isolates in Malaysia.

    Science.gov (United States)

    Tay, Sun Tee; Lotfalikhani, Azadeh; Sabet, Negar Shafiei; Ponnampalavanar, Sasheela; Sulaiman, Sofiah; Na, Shiang Ling; Ng, Kee Peng

    2014-10-01

    Candida nivariensis and C. bracarensis have been recently identified as emerging yeast pathogens which are phenotypically indistinguishable from C. glabrata. However, there is little data on the prevalence and antifungal susceptibilities of these species. This study investigated the occurrence of C. nivariensis and C. bracarensis in a culture collection of 185 C. glabrata isolates at a Malaysian teaching hospital. C. nivariensis was discriminated from C. glabrata using a PCR assay as described by Enache-Angoulvant et al. (J Clin Microbiol 49:3375-9, 2011). The identity of the isolates was confirmed by sequence analysis of the D1D2 domain and internal transcribed spacer region of the yeasts. The isolates were cultured on Chromogenic CHROMagar Candida (®) agar (Difco, USA), and their biochemical and enzymic profiles were determined. Antifungal susceptibilities of the isolates against amphotericin B, fluconazole, voriconazole and caspofungin were determined using E tests. Clotrimazole MICs were determined using a microbroth dilution method. There was a low prevalence (1.1 %) of C. nivariensis in our culture collection of C. glabrata. C. nivariensis was isolated from a blood culture and vaginal swab of two patients. C. nivariensis grew as white colonies on Chromogenic agar and demonstrated few positive reactions using biochemical tests. Enzymatic profiles of the C. nivariensis isolates were similar to that of C. glabrata. The isolates were susceptible to amphotericin B, fluconazole, voriconazole and caspofungin. Clotrimazole resistance is suspected in one isolate. This study reports for the first time the emergence of C. nivariensis in our clinical setting.

  3. Candida krusei and Candida glabrata reduce the filamentation of Candida albicans by downregulating expression of HWP1 gene.

    Science.gov (United States)

    de Barros, Patrícia Pimentel; Freire, Fernanda; Rossoni, Rodnei Dennis; Junqueira, Juliana Campos; Jorge, Antonio Olavo Cardoso

    2017-07-01

    Pathogenicity of Candida albicans is associated with its capacity switch from yeast-like to hyphal growth. The hyphal form is capable to penetrate the epithelial surfaces and to damage the host tissues. Therefore, many investigations have focused on mechanisms that control the morphological transitions of C. albicans. Recently, certain studies have showed that non-albicans Candida species can reduce the capacity of C. albicans to form biofilms and to develop candidiasis in animal models. Then, the objective of this study was to evaluate the effects of Candida krusei and Candida glabrata on the morphogenesis of C. albicans. Firstly, the capacity of reference and clinical strains of C. albicans in forming hyphae was tested in vitro. After that, the expression of HWP1 (hyphal wall protein 1) gene was determined by quantitative real-time PCR (polymerase chain reaction) assay. For both reference and clinical strains, a significant inhibition of the hyphae formation was observed when C. albicans was incubated in the presence of C. krusei or C. glabrata compared to the control group composed only by C. albicans. In addition, the culture mixed of C. albicans-C. krusei or C. albicans-C. glabrata reduced significantly the expression of HWP1 gene of C. albicans in relation to single cultures of this specie. In both filamentation and gene expression assays, C. krusei showed the higher inhibitory activity on the morphogenesis of C. albicans compared to C. glabrata. C. krusei and C. glabrata are capable to reduce the filamentation of C. albicans and consequently decrease the expression of the HWP1 gene.

  4. Dominant lethal effect of gamma radiation of 60Co in Biomphalaria glabrata (SAY, 1818)

    International Nuclear Information System (INIS)

    Tallarico, Lenita de Freitas

    2003-01-01

    Germ cell mutations are used in ecotoxicological studies as biomarkers of population effects and indicators of ecological changes. Biomphalaria glabrata, a freshwater mollusk, is a good experimental model for biomonitoring studies due to its biological characteristics and the ecological importance of this invertebrate group. The dominant lethal test was established in B. glabrata for the detection of germ cell mutations. Results with chemical mutagens showed that this system is efficient, specific and sensitive in the evaluation of germ cell mutations induced by reference mutagens. In this work, the dominant lethal effects of gamma radiation of 60 Co were studied. A preliminary experiment was done to establish the dose range and to estimate the chronology of spermatogenesis in B. glabrata. This estimate is possible because of the uniformity in response to ionizing radiation between germ cells at homologous stages of spermatogenesis in widely different species. In general, pre-meiotic germ cells are less sensitive to the induction of lethal dominant mutations than post-meiotic cells. This effect can be attributed to: young gametogenic cells - mitotically active - have greater repair ability from sub-lethal DNA damage and there is a selective elimination of the damaged cells. In our system: induction of lethal dominant mutations causes an increase in the frequency of malformations and, cytotoxic effect is displayed as a reduction in the crossing rates. Total duration of spermatogenesis was estimated in approximately 36 days, with the following distribution of stages: 1 to 13 days - spermatogonia, 14 to 20 days - spermatocytes, 21 to 36 days - spermatids and spermatozoa. Based on this chronology, irradiated wild-type snails with 2,5; 10 and 20Gy and crossed with non-irradiated albino snails after 7, 17, 23, 30 and 36 days. The frequencies of malformations in the heterozygous wild-type offspring of the nonirradiated albino snails were used as indicator of germ cell

  5. Growth of Biomphalaria glabrata populations in the presence of the ampullariid snails Pila ovata, Lanistes carinatus and Marisa cornuarietis.

    Science.gov (United States)

    Stryker, G A; Koech, D K; Loker, E S

    1991-06-01

    Adults of three ampullariid species were examined for their ability to affect the population growth of Biomphalaria glabrata (M line strain) under laboratory conditions in which food (lettuce) was supplied ad libitum over an eleven week interval. Lanistes carinatus and Pila ovata were obtained from Kenya, and Marisa cornuarietis originated from the Dominican Republic. The presence of either 3 or 9 L. carinatus or 3 M. cornuarietis per 40 l aquarium did not reduce the population size of B. glabrata below levels attained in control aquaria lacking ampullariids. The presence of 9 M. cornuarietis adults significantly reduced B. glabrata populations by week 10 of the experiment. Presence of 3 or 9 P. ovata per aquarium significantly reduced B. glabrata numbers below control levels starting at week four of the experiment. Aquaria with 9 P. ovata were observed to have fewer B. glabrata egg masses than control aquaria, but the number of egg masses observed was not significantly reduced in aquaria with 3 P. ovata or with 3 or 9 L. carinatus or M. cornuarietis. However, the presence of 3 or 9 adults of each ampullariid species resulted in a significant increase in the percentage of such egg masses that disappeared prior to expected hatching date and that were presumed to have been consumed. The results of his laboratory study suggest that further investigation of the role of ampullariids as biological control agents for pulmonate snails in sub-Saharan Africa should focus on P. ovata.

  6. Weight loss and survival of Biomphalaria Glabrata deprived of water

    Directory of Open Access Journals (Sweden)

    Marc Vianey-Liaud

    1986-06-01

    Full Text Available Immature and mature Biomphalaria glabrata are kept out of water at relative humidities varying from 0 to 100%. When snails are submitted to a saturated atmosphere, they show a slow weight loss and survival may be long. If relative humidity (RH decreases, weight loss becomes important and survival is short. A reduced RH (0 to 65% produces similar effects. During desiccation, fasting has no noticeable effect; survival depends essentially on weight loss.Biomphalaria glabrata maduros ou imaturos são mantidos fora da água, variando a umidade de 0 a 100%. Quando caramujos são submetidos a uma atmosfera saturada, sofrem uma lenta perda de peso e a sobrevivência pode ser longa. Se a umidade relativa decresce, a perda de peso será importante e a sobrevida será abreviada. Uma umidade relativa de 0 a 65% pode produzir efeitos similares. Durante a dessecação, a privação de alimento não tem efeito notável, a sobrevivência dependendo essencialmente da perda de peso.

  7. Silver colloidal nanoparticles: antifungal effect against adhered cells and biofilms of Candida albicans and Candida glabrata.

    Science.gov (United States)

    Monteiro, D R; Gorup, L F; Silva, S; Negri, M; de Camargo, E R; Oliveira, R; Barbosa, D B; Henriques, M

    2011-08-01

    The aim of this study was to evaluate the effect of silver nanoparticles (SN) against Candida albicans and Candida glabrata adhered cells and biofilms. SN (average diameter 5 nm) were synthesized by silver nitrate reduction with sodium citrate and stabilized with ammonia. Minimal inhibitory concentration (MIC) tests were performed for C. albicans (n = 2) and C. glabrata (n = 2) grown in suspension following the Clinical Laboratory Standards Institute microbroth dilution method. SN were applied to adhered cells (2 h) or biofilms (48 h) and after 24 h of contact their effect was assessed by enumeration of colony forming units (CFUs) and quantification of total biomass (by crystal violet staining). The MIC results showed that SN were fungicidal against all strains tested at very low concentrations (0.4-3.3 μg ml(-1)). Furthermore, SN were more effective in reducing biofilm biomass when applied to adhered cells (2 h) than to pre-formed biofilms (48 h), with the exception of C. glabrata ATCC, which in both cases showed a reduction ∼90%. Regarding cell viability, SN were highly effective on adhered C. glabrata and respective biofilms. On C. albicans the effect was not so evident but there was also a reduction in the number of viable biofilm cells. In summary, SN may have the potential to be an effective alternative to conventional antifungal agents for future therapies in Candida-associated denture stomatitis.

  8. Potential use of glycogen level as biomarker of chemical stress in Biomphalaria glabrata

    International Nuclear Information System (INIS)

    Ansaldo, Martin; Nahabedian, Daniel E.; Holmes-Brown, Eduardo; Agote, Marcos; Ansay, Cristina V.; Guerrero, Noemi R. Verrengia; Wider, Eva A.

    2006-01-01

    Biomphalaria glabrata, a freshwater gastropod mollusc, was tested as biondicator organism to assess cadmium, lead and arsenic exposure using acute laboratory bioassays. Modifications of glycogen levels were measured in different anatomical regions of B. glabrata in order to test the usefulness of this parameter as a general biomarker of chemical stress. The snails were exposed 96 h to different concentrations of the following contaminants: 0.1 and 0.05 mg Cd/L; 0.5, 0.1 and 0.05 mg Pb/L; 0.5, 0.1 and 0.05 mg As/L. Significant decreases in the polysaccharide content were observed in gonadal region for all treated animals. Arsenic and lead at 0.1 and 0.5 mg/L level of exposure were also able to decrease the levels of glycogen in the pulmonary and digestive gland region. Glycogen content in the cephalopedal region of treated animals presented a significant decrease (p < 0.05) when compared with control organisms only for arsenic at the highest level of exposure. To establish possible correlations between glycogen and contaminants accumulated by snails, analyses of the elements bioaccumulated in the different anatomical regions of B. glabrata were also performed. Cadmium and lead followed a similar pattern of bioaccumulation with highest values in the digestive gland region. Arsenic bioaccumulation, however, was highest in the gonadal region

  9. Formation of new chromosomes as a virulence mechanism in yeast Candida glabrata

    DEFF Research Database (Denmark)

    Poláková, S.; Blume, C.; Zárate, J. A.

    2009-01-01

    , Candida glabrata, for their genome structure and stability. This organism has recently become the second most prevalent yeast pathogen in humans. Although the gene sequences were well conserved among different strains, their chromosome structures differed drastically. The most frequent events reshaping...

  10. Influence of gamma radiation on the levels of polyphenols and lethality of ethanol extracts of Anacardium occidentale Linn., against Biomphalaria glabrata; Influencia da radiacao gama na acao moluscicida de extratos de cajueiro em Biomphalaria glabrata

    Energy Technology Data Exchange (ETDEWEB)

    Santos, G.H.F.; Silva, E.B., E-mail: santosghf@hotmail.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Energia Nuclear. Grupo de Radioprotecao e Radioecologia; Melo, A.M.M.A.; Lima, C.S.A [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de Biofisica; Amorim, E.L.C.; Peixoto Sobrinho, T.J.S. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Farmacia

    2013-08-15

    Plant materials rich in phenolic compounds, such as Anacardium occidentale Linn., Have been used as alternatives to synthetic pesticides in Biomphalaria glabrata control programs, intermediate host of Schistosoma mansoni. Studies show that ionizing radiation can influence the content of phenolic compounds and thus their biological actions. The aim of this study was to evaluate the influence of gamma radiation of {sup 60}Co in polyphenol composition of hydroalcoholic extracts of bark and leaves of A. occidentale and evaluate the toxicity of these extracts to embryos and adults of B. glabrata. To achieve this goal this, the extracts were irradiated at 10 kGy, the controls being maintained from 0 kGy and positive (CaCO{sub 3}) and negative (H{sub 2}O). We quantified the total phenols by the Folin-Ciocalteau and tannins by precipitation of casein. Extracts were used at a concentration of 100 mg/L. The results showed that the radiation caused the changes to the leaves, the percentage of polyphenols and tannins, and the percentage of lethality in embryos and adults Biomphalaria glabrata, these percentages being: 13 ± 5 (0 kGy) and 27 ± 2.5 (10 kGy), and 36.67 ± 5.77 (0 kGy), and 56.67 ± 5.77 (10 kGy), respectively. Gamma radiation caused significant changes in the levels of polyphenols in the extracts of leaves of Anacardium ocidentale Linn., translated by the increased toxicity of this extract against embryos and adults of Biomphalaria glabrata. This indicates that gamma radiation can be used as an agent potentiating the toxicity of plant extracts on the alternate use of these materials as molluscicides. (author)

  11. Towards New Antifolates Targeting Eukaryotic Opportunistic Infections

    Energy Technology Data Exchange (ETDEWEB)

    Liu, J.; Bolstad, D; Bolstad, E; Wright, D; Anderson, A

    2009-01-01

    Trimethoprim, an antifolate commonly prescribed in combination with sulfamethoxazole, potently inhibits several prokaryotic species of dihydrofolate reductase (DHFR). However, several eukaryotic pathogenic organisms are resistant to trimethoprim, preventing its effective use as a therapeutic for those infections. We have been building a program to reengineer trimethoprim to more potently and selectively inhibit eukaryotic species of DHFR as a viable strategy for new drug discovery targeting several opportunistic pathogens. We have developed a series of compounds that exhibit potent and selective inhibition of DHFR from the parasitic protozoa Cryptosporidium and Toxoplasma as well as the fungus Candida glabrata. A comparison of the structures of DHFR from the fungal species Candida glabrata and Pneumocystis suggests that the compounds may also potently inhibit Pneumocystis DHFR.

  12. Ionotropic Receptors Identified within the Tentacle of the Freshwater Snail Biomphalaria glabrata, an Intermediate Host of Schistosoma mansoni.

    Directory of Open Access Journals (Sweden)

    Di Liang

    Full Text Available Biomphalaria glabrata (B. glabrata is an air-breathing aquatic mollusc found in freshwater habitats across the Western Hemisphere. It is most well-known for its recognized capacity to act as a major intermediate host for Schistosoma mansoni, the human blood fluke parasite. Ionotropic receptors (IRs, a variant family of the ionotropic glutamate receptors (iGluR, have an evolutionary ancient function in detecting odors to initiate chemosensory signaling. In this study, we applied an array of methods towards the goal of identifying IR-like family members in B. glabrata, ultimately revealing two types, the iGluR and IR. Sequence alignment showed that three ligand-binding residues are conserved in most Biomphalaria iGluR sequences, while the IRs did exhibit a variable pattern, lacking some or all known glutamate-interactingresidues, supporting their distinct classification from the iGluRs. We show that B. glabrata contains 7 putative IRs, some of which are expressed within its chemosensory organs. To further investigate a role for the more ancient IR25a type in chemoreception, we tested its spatial distribution pattern within the snail cephalic tentacle by in situ hybridization. The presence of IR25a within presumptive sensory neurons supports a role for this receptor in olfactory processing, contributing to our understanding of the molecular pathways that are involved in Biomphalaria olfactory processing.

  13. Contribution to the histology of Biomphalaria glabrata Contribuição à histologia da Biomphalaria glabrata

    Directory of Open Access Journals (Sweden)

    Queli Teixeira Lemos

    1999-08-01

    Full Text Available A combination of histological techniques applied to the study of Biomphalaria glabrata yielded some interesting new data about the histology of this snail, a major intermediate host of Schistosoma mansoni in Brazil. Three kinds of pigments were identified: a dark pigment which bleached following oxidation with potassium permanganate; a lipofuchsin-like, diastase-resistant PAS-positive pigment and an iron-containing pigment, probably related to hemosiderin. Calcium was detected in small deposits within the connective tissue and forming a dense core inside the chitinous radular teeth. The presence of fibrils, staining with sirius-red and birefringence under polarized light strongly suggest primitive collagen tissue. The radular apparatus appeared as a storing site for glycogen, while abundant Alcian-blue positive material (proteoglycans was extremely concentrated in the radular sac.Uma combinação de várias técnicas histológicas permitiu alguns achados de interesse para a histologia da Biomphalaria glabrata, principal hospedeiro intermediário do Schistosoma mansoni no Brasil. Três tipos de pigmentos foram identificados: um pigmento escuro, que se descora após oxidação pelo permanganato de potássio; outro que é semelhante à lipofuscina, que se cora pelo PAS e é diastase- resistente, e um pigmento que contém ferro, provalvelmente, relacionado com a hemossiderina. O cálcio foi identificado em pequenos depósitos no interior do tecido conjuntivo e formando pequenos núcleos no interior dos dentes quitinosos da rádula. A presença de fibrilas coradas pelo sírius-vermelho, exibindo birrefrigência sob luz polarizada, sugere fortemente a presença de um tecido colágeno primitivo. O aparelho radular foi visto como um local de armazenamento do glicogênio, enquanto a presença de abundante material amorfo positivo para o azul de alciano (proteoglicano apareceu em grande concentração no saco radular.

  14. Chronic vulvovaginal candidiasis: characteristics of women with Candida albicans, C glabrata and no candida.

    Science.gov (United States)

    Geiger, A M; Foxman, B; Sobel, J D

    1995-01-01

    INTRODUCTION--Although as many as 5% of all women complain of chronic vulvovaginitis, little is known about these women. They may often be misdiagnosed and the role of vaginal yeast culture in diagnosing vulvovaginal candidiasis (VVC) among them has not been clearly defined. METHODS--To address these deficiencies, we tabulated initial diagnoses among new patients and conducted a medical record-based, unmatched case-control study among women reporting a history of chronic vulvovaginitis (four or more episodes in the past year) at a vulvovaginitis specialty clinic. Clinical presentation and medical history were compared for women who had a positive vaginal yeast culture for either Candida albicans or C glabrata, or who had a negative culture. RESULTS--One-third of the women had no apparent vulvovaginal disease at their initial visit. All women reported similar symptoms, except for an increased prevalence of painful sexual intercourse in women with C albicans (chi 2 p = 0.014 versus women with C glabrata and p vulvovaginitis. Among women with VVC, subtle differences in clinical presentation do not reliably distinguish women with C albicans from those with C glabrata. Our study also indicates that vaginal yeast cultures, while not necessary for every patient, are valuable in confirming negative diagnoses, detecting microscopy false-negatives, and identifying non-C albicans isolates. Images PMID:7490047

  15. Infectivity of Trichinella papuae for experimentally infected red foxes (Vulpes vulpes)

    DEFF Research Database (Denmark)

    Webster, P.; Malakauskas, A.; Kapel, C. M O

    2002-01-01

    To evaluate infectivity for carnivores as well as other biological characteristics of the newly described Trichinella papuae, eight red foxes were experimentally infected with the parasite. Five weeks after inoculation, T. papuae larvae were recovered from nine different muscle types. The larvae...

  16. Interactions between Candida albicans and Candida glabrata in biofilms: Influence of the strain type, culture medium and glucose supplementation.

    Science.gov (United States)

    Hosida, Thayse Yumi; Cavazana, Thamires Priscila; Henriques, Mariana; Pessan, Juliano Pelim; Delbem, Alberto Carlos Botazzo; Monteiro, Douglas Roberto

    2018-04-01

    The relationship among Candida species may be influenced by several factors. Thus, this study evaluated the interactions between Candida albicans and Candida glabrata in biofilms, varying the strain type, culture medium and glucose supplementation. Biofilms were formed for 48 hours in Sabouraud dextrose broth (SDB) or RPMI 1640, supplemented with 0%, 1% or 5% glucose. Each strain of C. albicans was combined with two strains of C. glabrata, generating four biofilm associations, which were quantified by colony-forming units (CFUs), total biomass and metabolic activity. Data were analysed by ANOVA and Tukey's HSD test (α = 0.05). For CFUs, all associations were classified as indifferent for biofilms formed in RPMI 1640, while for SDB the interactions were antagonistic for C. albicans and indifferent for C. glabrata. The association of reference strains resulted in a dual-species biofilm with biomass significantly higher than that observed for each single biofilm developed in SDB. The metabolic activity of dual-species biofilms did not significantly differ from that found for single ones, except for co-culture of the reference strains. Glucose supplementation and culture media had a significant influence on all parameters. In conclusion, the strain type, culture medium and glucose supplementation influenced the interactions between C. albicans and C. glabrata. © 2017 Blackwell Verlag GmbH.

  17. Candida glabrata tryptophan-based pigment production via the Ehrlich pathway.

    Science.gov (United States)

    Brunke, Sascha; Seider, Katja; Almeida, Ricardo Sergio; Heyken, Antje; Fleck, Christian Benjamin; Brock, Matthias; Barz, Dagmar; Rupp, Steffen; Hube, Bernhard

    2010-04-01

    Pigments contribute to the pathogenicity of many fungi, mainly by protecting fungal cells from host defence activities. Here, we have dissected the biosynthetic pathway of a tryptophan-derived pigment of the human pathogen Candida glabrata, identified key genes involved in pigment production and have begun to elucidate the possible biological function of the pigment. Using transcriptional analyses and a transposon insertion library, we have identified genes associated with pigment production. Targeted deletion mutants revealed that the pigment is a by-product of the Ehrlich pathway of tryptophan degradation: a mutant lacking a tryptophan-upregulated aromatic aminotransferase (Aro8) displayed significantly reduced pigmentation and a recombinantly expressed version of this protein was sufficient for pigment production in vitro. Pigment production is tightly regulated as the synthesis is affected by the presence of alternative nitrogen sources, carbon sources, cyclic AMP and oxygen. Growth of C. glabrata on pigment inducing medium leads to an increased resistance to hydrogen peroxide, an effect which was not observed with a mutant defective in pigmentation. Furthermore, pigmented yeast cells had a higher survival rate when exposed to human neutrophils and caused increased damage in a monolayer model of human epithelia, indicating a possible role of pigmentation during interactions with host cells.

  18. Oxidative stress in mollusks Biomphalaria Glabrata exposed to gamma radiation

    International Nuclear Information System (INIS)

    Silva, Luanna R.S.; Augusto, Ricielle L.; Siqueira, Williams N.; Luna Filho, Ricardo L.C.; Pereira, Dewson R.; França, Elvis J.; Silva, Edvane B.; Melo, Ana M.M.A.

    2017-01-01

    Ionizing radiation can cause biological changes in different organisms such as mollusks from Biomphalaria glabrata species, in which alterations could be observed in the reproductive system of the specimens, prejudicing fertility and fecundity. As the changes may occur due to the lipid peroxidation caused by the action of free radicals on the gonads, the objective of this work was to evaluate the oxidative damage caused by the exposure of B. glabrata mollusks to different doses of gamma radiation. In addition, efforts were carried out to standardize a sensitive and low-cost technique for detecting negative effects caused by high doses of ionizing radiation. For this, each mollusk group (n = 10) was submitted to 0 (control), 10, 15, 20 and 25 Gy (Gammacell- "6"0Co, dose rate 3.532 kGy/h). The TBARS method was applied for the quantification of lipid peroxidation of the gonads of the mollusks after 24 and 48 h. ANOVA, followed by the mean comparison (Tukey) at the 5% of significance level, indicated high concentrations of TBARS in the gonads after 24 h. Otherwise, after 48 h, differences for TBARS concentrations were not significant at the 95% confidence level, determining that the action of free radicals from ionizing radiation on cell membranes mainly occurred within 24 h after irradiation. (author)

  19. Oxidative stress in mollusks Biomphalaria Glabrata exposed to gamma radiation

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Luanna R.S.; Augusto, Ricielle L.; Siqueira, Williams N.; Luna Filho, Ricardo L.C.; Pereira, Dewson R.; França, Elvis J.; Silva, Edvane B.; Melo, Ana M.M.A., E-mail: luannaribeiro_lua@hotmail.com, E-mail: williams.wns@gmail.com, E-mail: ejfranca@gmail.com, E-mail: edvborges@yahoo.com, E-mail: luannaribeiro_lua@hotmail.com, E-mail: williams.wns@gmail.com, E-mail: ricardolclf@hotmail.com, E-mail: dewson.rocha@gmail.com, E-mail: s, E-mail: amdemelo@hotmail.com, E-mail: ricielleaugusto@gmail.com, E-mail: williams.wns@gmail.com, E-mail: ejfranca@gmail.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil); Centro Regional de Ciencias Nucleares do Nordeste (CRCN-NE/CNEN-PE), Recife, PE (Brazil)

    2017-11-01

    Ionizing radiation can cause biological changes in different organisms such as mollusks from Biomphalaria glabrata species, in which alterations could be observed in the reproductive system of the specimens, prejudicing fertility and fecundity. As the changes may occur due to the lipid peroxidation caused by the action of free radicals on the gonads, the objective of this work was to evaluate the oxidative damage caused by the exposure of B. glabrata mollusks to different doses of gamma radiation. In addition, efforts were carried out to standardize a sensitive and low-cost technique for detecting negative effects caused by high doses of ionizing radiation. For this, each mollusk group (n = 10) was submitted to 0 (control), 10, 15, 20 and 25 Gy (Gammacell- {sup 60}Co, dose rate 3.532 kGy/h). The TBARS method was applied for the quantification of lipid peroxidation of the gonads of the mollusks after 24 and 48 h. ANOVA, followed by the mean comparison (Tukey) at the 5% of significance level, indicated high concentrations of TBARS in the gonads after 24 h. Otherwise, after 48 h, differences for TBARS concentrations were not significant at the 95% confidence level, determining that the action of free radicals from ionizing radiation on cell membranes mainly occurred within 24 h after irradiation. (author)

  20. Effect of cadmium, lead and arsenic on the oviposition, hatching and embryonic survival of Biomphalaria glabrata

    International Nuclear Information System (INIS)

    Ansaldo, Martin; Nahabedian, Daniel E.; Di Fonzo, Carla; Wider, Eva A.

    2009-01-01

    Biomphalaria glabrata is a widespread freshwater gastropod mollusc. The easy aquaculture of these organisms allow its use as an accessible tool for contamination bioassays. B. glabrata showed marked metabolic responses when exposed to cadmium, lead and arsenic. Those responses could also affect the reproduction of the snails. Taking into account this hypothesis, B. glabrata were exposed for 96 h (acute laboratory bioassays) to different concentrations of cadmium (0.1, 0.05 and 0 mg/L), lead (0.5, 0.1, 0.05 and 0 mg/L) and arsenic (0.5, 0.1, 0.05 and 0 mg/L). Snails were removed from the aquaria while eggs were left in the same contaminant concentrations. The effect of the assayed toxicants on snail reproduction was registered as the alterations of the total number of laid eggs (TNLE), hatching time and embryonic survival. At 0.10 mg/L cadmium significantly decreased the TNLE (p < 0.05) and no embryos survived. The lowest assayed level (0.05 mg/L) of cadmium, delayed the hatching time twice when it was compared with the control group (p < 0.01). Lead decreased the TNLE at 0.5 mg/L level (p < 0.01). The other assayed doses (0.05 and 0.10 mg/L) also decreased embryonic survival significantly (p < 0.05 and p < 0.01 respectively) and extended twice the time to hatching (p < 0.01). The 0.50 mg/L level killed all embryos. Arsenic at all studied concentrations decreased the TNLE (p < 0.05) while the hatching time was increased by 50%. Embryo survival only decreased at the highest level (0.5 mg/L) of arsenic assayed. In summary, the acute exposure (96 h) to cadmium lead and arsenic, altered the reproduction of B. glabrata, modifying the TNLE, hatching time and embryonic survival

  1. Atividade moluscicida de princípios ativos de folhas de Lycopersicon esculentum (Solanales, Solanaceae em Biomphalaria glabrata (Gastropoda, Planorbidae Moluscicide activity of active principles in the leaves of Lycopersicon esculentum (Solanales, Solanaceae on Biomphalaria glabrata (Gastropoda, Planorbidae

    Directory of Open Access Journals (Sweden)

    Vilma Leyton

    Full Text Available Foram obtidos extratos aquosos e alcoólicos a partir de pó de folhas secas de tomateiro (Lycopersicon esculentum, Mill. c.v. Cereja. Por extração metanólica e precipitação alcalina, foi obtido um produto que denominamos "glicoalcalóide esteroidal bruto" (GEb, no qual foi caracterizada a presença de tomatina. Em ensaios laboratoriais, os extratos aquosos, alcoólicos e o GEb apresentaram atividade moluscicida em Biomphalaria glabrata (Say, 1818. O "glicoalcalóide esteroidal bruto" apresentou alta atividade moluscicida (CL50 = 8,01 ppm e CL90 = 13,17 ppm, comparável à atividade da tomatina. Desovas de B. glabrata mostraram-se resistentes aos extratos testados. Os níveis de atividade moluscicida apresentados pelos diversos extratos e o GEb, apontam apenas o GEb como candidato para a continuação dos estudos visando a sua possível utilização em campo.Aqueous and alcoholic extracts were obtained from crushed dried leaves of tomato plant (Lycopersicon esculentum, Mill. c.v. Cherry. By the use of a methanolic extraction and alkaline precipitation, a product named crude steroidal glycoalkaloid (GEb, was obtained. The presence of tomatidine was characterized in this product. In laboratory, the aqueous and alcoholic extracts and GEb have shown molluscicidal activity against Biomphalaria glabrata (Say, 1818. The crude steroidal glycoalkaloid presented a high molluscicidal activity (LC50 = 8.01 ppm and LC90 = 13.17 ppm, similar to that of tomatine. None of the compounds tested affected B. glabrata egg masses. The level of activity showed by the different extracts and by the GEb, pointed out the GEb as the only candidate able to be considered for further tests toward field trials as molluscicidal agent.

  2. β-Glucan induces reactive oxygen species production in human neutrophils to improve the killing of Candida albicans and Candida glabrata isolates from vulvovaginal candidiasis.

    Directory of Open Access Journals (Sweden)

    Patricia de Souza Bonfim-Mendonça

    Full Text Available Vulvovaginal candidiasis (VVC is among the most prevalent vaginal diseases. Candida albicans is still the most prevalent species associated with this pathology, however, the prevalence of other Candida species, such as C. glabrata, is increasing. The pathogenesis of these infections has been intensely studied, nevertheless, no consensus has been reached on the pathogenicity of VVC. In addition, inappropriate treatment or the presence of resistant strains can lead to RVVC (vulvovaginal candidiasis recurrent. Immunomodulation therapy studies have become increasingly promising, including with the β-glucans. Thus, in the present study, we evaluated microbicidal activity, phagocytosis, intracellular oxidant species production, oxygen consumption, myeloperoxidase (MPO activity, and the release of tumor necrosis factor α (TNF-α, interleukin-8 (IL-8, IL-1β, and IL-1Ra in neutrophils previously treated or not with β-glucan. In all of the assays, human neutrophils were challenged with C. albicans and C. glabrata isolated from vulvovaginal candidiasis. β-glucan significantly increased oxidant species production, suggesting that β-glucan may be an efficient immunomodulator that triggers an increase in the microbicidal response of neutrophils for both of the species isolated from vulvovaginal candidiasis. The effects of β-glucan appeared to be mainly related to the activation of reactive oxygen species and modulation of cytokine release.

  3. Blood biochemistry responses of chickens experimentally infected ...

    African Journals Online (AJOL)

    This study investigated the blood biochemistry responses of cockerels experimentally infected with a velogenic Newcastle disease virus (NDV) strain, KUDU 113. One hundred Isa white cockerels were used for the study. The cockerels were obtained at day-old and randomly divided into groups A- vaccinated and infected, ...

  4. Nitric oxide production by Biomphalaria glabrata haemocytes: effects of Schistosoma mansoni ESPs and regulation through the extracellular signal-regulated kinase pathway

    Directory of Open Access Journals (Sweden)

    Kirk Ruth S

    2009-04-01

    Full Text Available Abstract Background Schistosoma mansoni uses Biomphalaria glabrata as an intermediate host during its complex life cycle. In the snail, the parasite initially transforms from a miracidium into a mother sporocyst and during this process excretory-secretory products (ESPs are released. Nitric oxide (NO and its reactive intermediates play an important role in host defence responses against pathogens. This study therefore aimed to determine the effects of S. mansoni ESPs on NO production in defence cells (haemocytes from schistosome-susceptible and schistosome-resistant B. glabrata strains. As S. mansoni ESPs have previously been shown to inhibit extracellular signal-regulated kinase (ERK phosphorylation (activation in haemocytes from susceptible, but not resistant, B. glabrata the regulation of NO output by ERK in these cells was also investigated. Results Haemocytes from resistant snails challenged with S. mansoni ESPs (20 μg/ml over 5 h displayed an increase in NO production that was 3.3 times greater than that observed for unchallenged haemocytes; lower concentrations of ESPs (0.1–10 μg/ml did not significantly increase NO output. In contrast, haemocytes from susceptible snails showed no significant change in NO output following challenge with ESPs at any concentration used (0.1–20 μg/ml. Western blotting revealed that U0126 (1 μM or 10 μM blocked the phosphorylation (activation status of ERK in haemocytes from both snail strains. Inhibition of ERK signalling by U0126 attenuated considerably intracellular NO production in haemocytes from both susceptible and resistant B. glabrata strains, identifying ERK as a key regulator of NO output in these cells. Conclusion S. mansoni ESPs differentially influence intracellular NO levels in susceptible and resistant B. glabrata haemocytes, possibly through modulation of the ERK signalling pathway. Such effects might facilitate survival of S. mansoni in its intermediate host.

  5. Estratégia competitiva entre Biomphalaria glabrata e Biomphalaria tenagophila: estudos de laboratório

    Directory of Open Access Journals (Sweden)

    Mairy Barbosa Loureiro dos Santos

    1989-01-01

    Full Text Available Observation about conhabitation among B. glabrata and B. tenagophila revealed a greater vulnerability of B. tenagophila population during the process of competition when its density was severaly decreased in 12 trials, moderate in 2 trials. It was higher than B. glabrata in only one trial. Some snail water chemical parameters analysed such as pH, alkalinity, conductivity and oxygen dissolve, an the viability rate of batch of eggs didn't give subsidy to explain the competition mechanism. The newly-born survival, in the situation of cohabitation, was low for both species. This reveals the existence of intra and interspecific competitive interacition. The fertility rate reduction of B. tenagophila during the cohabitation was considered as a cause of its exlusion. One of the factors that seems to have influenced the fertility rate was a possible wrong crossing.

  6. Antiestrogenic constituents of the Thai medicinal plants Capparis flavicans and Vitex glabrata.

    Science.gov (United States)

    Luecha, Prathan; Umehara, Kaoru; Miyase, Toshio; Noguchi, Hiroshi

    2009-11-01

    Antiestrogenic compounds were investigated from Thai indigenous plants for galactogogues since estrogen is reported to suppress lactation in breastfeeding women. The aerial parts of the Thai medicinal plant Capparis flavicans, which has traditionally been used to promote lactation, gave the new compound capparoside A (1), along with 28 known compounds. The leaves of Vitex glabrata belong to the same genus as the chaste tree (Vitex agnus-castus), which is used traditionally to support lactation, and afforded the new compounds khainaoside A (14), khainaoside B (15), and khainaoside C (16), together with six known compounds. The isolates were tested for their biological activity using the estrogen-responsive human breast cancer cell lines MCF-7 and T47D. Syringaresinol (3) and principin (6), from C. flavicans, and khainaoside A (14) showed the most potent inhibitory effects on estrogen-enhanced cell proliferation among all compounds isolated. These results suggest that the lactation-promoting properties of C. flavicans might be related to the inhibitory effect on excess estrogen of women who experience insufficient breastfeeding and highlight the possibility of using V. glabrata leaves for their antiestrogenic properties.

  7. The Compatibility Between Biomphalaria glabrata Snails and Schistosoma mansoni: An Increasingly Complex Puzzle.

    Science.gov (United States)

    Mitta, G; Gourbal, B; Grunau, C; Knight, M; Bridger, J M; Théron, A

    2017-01-01

    This review reexamines the results obtained in recent decades regarding the compatibility polymorphism between the snail, Biomphalaria glabrata, and the pathogen, Schistosoma mansoni, which is one of the agents responsible for human schistosomiasis. Some results point to the snail's resistance as explaining the incompatibility, while others support a "matching hypothesis" between the snail's immune receptors and the schistosome's antigens. We propose here that the two hypotheses are not exclusive, and that the compatible/incompatible status of a particular host/parasite couple probably reflects the balance of multiple molecular determinants that support one hypothesis or the other. Because these genes are involved in a coevolutionary arms race, we also propose that the underlying mechanisms can vary. Finally, some recent results show that environmental factors could influence compatibility. Together, these results make the compatibility between B. glabrata and S. mansoni an increasingly complex puzzle. We need to develop more integrative approaches in order to find targets that could potentially be manipulated to control the transmission of schistosomiasis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Radioprotective effect of the extract of Ziziphus joazeiro and Anacardium occidentale on embryos of Biomphalaria glabrata submitted to ionizing radiation

    International Nuclear Information System (INIS)

    Siqueira, Williams N.; Silva, Luanna R.S.; Silva, Edvane B.; Silva, Ronaldo C.; Lacerda, Laila B.N.; Silva, Hianna A.M.F.; Santos, Mariana L.O.; Sa, Jose L.F.; Melo, Ana M.M.A.

    2011-01-01

    Electromagnetic radiations are energies that can be classified as non-ionizing and ionizing. This type of energy is propagated by a material medium and the vacuum. The important characteristic of ionizing radiation is the localized release of large amounts of energy. The biological effects of radiation result principally from damage to DNA, which is the critical target. Given these harmful effects caused by radiation highlights the importance of acquiring knowledge about the radioprotective substance, because they act to protect the living tissue, decreasing the damage he caused by the effects of radiation. In this study we investigated the radioprotective effect of extract hydroalcoholic of Ziziphus joazeiro and Anacardium occidentale on embryos of Biomphalaria glabrata. The embryos of Biomphalaria glabrata pigmented were divided into 18 groups of 100 specimens. The experimental groups were exposed to the extracts at a concentration of 200 ppm and then irradiated. For irradiation, we used a source of 60 Co (Gammacell of Radionics Labs. Dose rate = 4.359 Gy/h). The viability of the embryos was examined using a stereoscopic microscope and statistical analysis was performed using the test Student-Newman-Keuls and χ 2 . Our results showed that the extracts of hydroalcoholic Ziziphus joazeiro showed radioprotective effect and that the aqueous extract of the bark of Anacardium occidentale exhibited a reduction in its embryotoxic effect. (author)

  9. Radioprotective effect of the extract of Ziziphus joazeiro and Anacardium occidentale on embryos of Biomphalaria glabrata submitted to ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Siqueira, Williams N.; Silva, Luanna R.S.; Silva, Edvane B. [Universidade Federal de Pernambuco (DEN/UFPE), Recife, PE (Brazil). Dept. de Energia Nuclear. Grupo de Estudos em Radioprotecao e Radioecologia; Silva, Ronaldo C. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de Genetica; Lacerda, Laila B.N.; Silva, Hianna A.M.F.; Santos, Mariana L.O.; Sa, Jose L.F.; Melo, Ana M.M.A. [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Dept. de de Biofisica e Radiobiologia. Lab. de Radiobiologia

    2011-07-01

    Electromagnetic radiations are energies that can be classified as non-ionizing and ionizing. This type of energy is propagated by a material medium and the vacuum. The important characteristic of ionizing radiation is the localized release of large amounts of energy. The biological effects of radiation result principally from damage to DNA, which is the critical target. Given these harmful effects caused by radiation highlights the importance of acquiring knowledge about the radioprotective substance, because they act to protect the living tissue, decreasing the damage he caused by the effects of radiation. In this study we investigated the radioprotective effect of extract hydroalcoholic of Ziziphus joazeiro and Anacardium occidentale on embryos of Biomphalaria glabrata. The embryos of Biomphalaria glabrata pigmented were divided into 18 groups of 100 specimens. The experimental groups were exposed to the extracts at a concentration of 200 ppm and then irradiated. For irradiation, we used a source of {sup 60}Co (Gammacell of Radionics Labs. Dose rate = 4.359 Gy/h). The viability of the embryos was examined using a stereoscopic microscope and statistical analysis was performed using the test Student-Newman-Keuls and {chi}{sup 2}. Our results showed that the extracts of hydroalcoholic Ziziphus joazeiro showed radioprotective effect and that the aqueous extract of the bark of Anacardium occidentale exhibited a reduction in its embryotoxic effect. (author)

  10. Bird attributes, plant characteristics, and seed dispersal of Pera glabrata (Schott, 1858), (Euphorbiaceae) in a disturbed cerrado area.

    Science.gov (United States)

    Francisco, M R; Lunardi, V O; Galetti, M

    2007-11-01

    Several plant characteristics, such as fruit production, nutrient reward, secondary compounds, and fruit color display, affect fruit choice by birds. On the other hand, several bird attributes affect their efficiency as dispersers. Here we investigate the ornithochoric seed dispersal of Pera glabrata Schott (Euphorbiaceae) in a cerrado fragment in southeastern Brazil. A set of bird attributes, such as frequency of visits, number of diaspores eaten, time spent foraging, methods of taking and handling the diaspores and agonistic interactions were analyzed in order to infer about the potential of each species to act as a seed disperser. Birds were the unique seed dispersers of these oil-rich diaspores. We observed 414 bird visits during 60 hours of focal observations in five trees from December 1999 to January 2000. Twenty bird species from seven families ate the diaspores of P. glabrata, but only 14 species were considered potential seed dispersers because they swallowed the diaspores, increasing the probabilities for the seeds to be defecated and/or regurgitated away from the parent trees. The main potential seed dispersers were: Turdus leucomelas (Muscicapidae), Dacnis cayana (Emberizidae), Colaptes melanochloros (Picidae) and Elaenia spp. (Tyrannidae). We did not find any significant seasonal change in the number of visits on the fruiting trees throughout the day. We also did not find any relation between the number of visits per tree and fruit production. The most effective seed dispersers of P. glabrata were generalist birds, which have a high visiting rate, high fruit consumption rate, and spend short periods on the plants. The large number of species recorded as potential seed dispersers of P. glabrata, being most of them very abundant even in Brazilian disturbed areas, may guarantee seed dispersal of this plant in small fragments and regenerating areas.

  11. Suscetibilidade de biomphalaria glabrata, B. straminea e B. tenagophila a diferentes cepas de schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Luiz Candido de Souza Dias

    1987-08-01

    Full Text Available Em condições experimentais foi estudada a suscetibilidade de Biomphalaria glabrata, B. straminea e B. tenagophila a quatro linhagens humanas (MAP, PTH, UPH, e OuH e duas de roedores silvestres (PTR e VPR do Schistosoma mansoni. Grupos de 50 moluscos foram expostos individualmente a 10 miracídios e observados durante 70 dias. Avaliou-se a suscetibilidade dos moluscos ao parasito por meio da % de animais com esporocistos, % de moluscos que eliminavam cercárias e mortalidade conjunta dos animais expostos e infectados. Exemplares de B. glabrata mineira infectaram-se com cepa simpátrica (MAP e com 5 alopátricas do Estado de São Paulo (PTH, VPH, OuH, PTR e VPR. B. glabrata paulista mostrou altas taxas de infecção com as cepas MAP, VPR e OuH do trematódeo. Quatro % dos exemplares B. straminea de São Paulo eliminavam cercárias de cepas simpátricas; com cepa mineira apenas 4% apresentaram esporocistos na vigência de 20 miracídios por molusco; as menores taxas de mortalidade foram registradas com essa espécie de molusco, não sendo maior do que 20%. B. tenagophila paulista foi suscetível apenas às linhagens simpáticas sendo 6% a maior taxa de moluscos que eliminaram cercárias. Os resultados indicam que os movimentos populacionais humanos dentro do território paulista e para fora dele são importantes na disseminação da esquistossomose mansônica.

  12. Infections in orthopaedic surgery : clinical and experimental studies

    NARCIS (Netherlands)

    Vogely, Henri Charles

    2000-01-01

    The diagnostic difficulties, variability in outcome and the heterogeinity of the problem of orthopaedic infections stimulated the author to a study of the literature, and several clinical and experimental studies. The diagnosis prosthesis-related infection can only be reached with an acceptable

  13. Evaluation of radiosensitivity hemocytes of Biomphalaria glabrata exposed to gamma radiation

    International Nuclear Information System (INIS)

    Silva, L.R.S.; Amaral, A.J.; Silva, E.B.; Amancio, F.F.; Melo, A.M.M.A.

    2013-01-01

    The mollusc Biomphalaria glabrata have characteristics that allow them to be identified as an animal model ideal for monitoring areas exposed to chemical agents and physical. This study evaluated the effect of ionizing radiation from Cobalt-60 in haemocytes present in the hemolymph of Biomphalaria glabrata, with the goal of using these cells as indicators of the presence of radiation in aquatic environments. The mollusks were divided into five groups: one control and four subjected doses of 25, 35, 45 and 55 Gy of gamma radiation. After 48 hours of irradiation, the clam hemolymph was collected and slides were prepared and stained with Giemsa for analyses under a light microscope. Statistical analysis was performed using ANOVA and Tukey's test, p <0.05. The results showed that the total number of cells after irradiation reduced compared to control except at a dose of 55 Gy. During data analysis, morphological changes were observed in haemocytes of mollusks subjected to doses of 35, 45 and 55 Gy. These modifications consisted of nucleus bilobulated and nucleo plasmatic bridges. Another change was exclusively observed in the cellular exposure of 55 Gy, where hemocytes showed misshapen nuclei and cytoplasm vacuolisation, suggestive of apoptosis. It is concluded that hemocytes are sensitive to radiation and can be used as indicators of the presence of high doses of ionizing radiation in aquatic environments. (author)

  14. Clinical characteristics of the first cases of invasive candidiasis in China due to pan-echinocandin-resistant Candida tropicalis and Candida glabrata isolates with delineation of their resistance mechanisms

    Directory of Open Access Journals (Sweden)

    Xiao M

    2018-01-01

    Full Text Available Meng Xiao,1,2,* Xin Fan,1–3,* Xin Hou,1,2,4,* Sharon CA Chen,5 He Wang,1,2 Fanrong Kong,5 Zi-Yong Sun,6,* Yun-Zhuo Chu,7 Ying-Chun Xu1,2 1Department of Clinical Laboratory, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 2Beijing Key Laboratory for Mechanisms Research and Precision Diagnosis of Invasive Fungal Diseases, Beijing, People’s Republic of China; 3Department of Infectious Diseases and Clinical Microbiology, Beijing Chaoyang Hospital, Beijing, People’s Republic of China; 4Graduate School, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 5Center for Infectious Diseases and Microbiology Laboratory Services, ICPMR, New South Wales Health Pathology, Westmead Hospital, The University of Sydney, NSW, Australia; 6Department of Clinical Laboratory, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 7Department of Clinical Laboratory, The First Affiliated Hospital of Chinese Medical University, Shenyang, People’s Republic of China *These authors contributed equally to this work Abstract: Echinocandin antifungal agents have become the first-line therapy for invasive candidiasis (IC in many countries. Despite their increasing use, resistance to this class of drug is, overall, still uncommon. Here, we report two patients from the People’s Republic of China with IC, one with infection caused by pan-echinocandin-resistant Candida tropicalis and the other by pan-echinocandin-resistant Candida glabrata. We also describe the mechanisms of drug resistance of these isolates. The echinocandin-resistant C. glabrata isolate was cultured from ascitic fluid of a 46-year-old male patient with intra-abdominal IC developing after surgery in 2012. This patient had had no prior antifungal exposure. The echinocandin-resistant C. tropicalis isolate was cultured from chest drainage

  15. Quantification of HTLV-I proviral load in experimentally infected rabbits

    Directory of Open Access Journals (Sweden)

    Kindt Thomas J

    2005-05-01

    Full Text Available Abstract Background Levels of proviral load in HTLV-1 infected patients correlate with clinical outcome and are reasonably prognostic. Adaptation of proviral load measurement techniques is examined here for use in an experimental rabbit model of HTLV-1 infection. Initial efforts sought to correlate proviral load with route and dose of inoculation and with clinical outcome in this model. These methods contribute to our continuing goal of using the model to test treatments that alleviate virus infection. Results A real-time PCR assay was used to measure proviral load in blood and tissue samples from a series of rabbits infected using HTLV-1 inocula prepared as either cell-free virus particles, infected cells or blood, or by naked DNA injection. Proviral loads from asymptomatically infected rabbits showed levels corresponding to those reported for human patients with clinically silent HTLV-1 infections. Proviral load was comparably increased in 50% of experimentally infected rabbits that developed either spontaneous benign or malignant tumors while infected. Similarly elevated provirus was found in organs of rabbits with experimentally induced acute leukemia/lymphoma-like disease. Levels of provirus in organs taken at necropsy varied widely suggesting that reservoirs of infections exist in non-lymphoid organs not traditionally thought to be targets for HTLV-1. Conclusion Proviral load measurement is a valuable enhancement to the rabbit model for HTLV-1 infection providing a metric to monitor clinical status of the infected animals as well as a means for the testing of treatment to combat infection. In some cases proviral load in blood did not reflect organ proviral levels, revealing a limitation of this method for monitoring health status of HTLV-1 infected individuals.

  16. Molluscicidal activity of Manilkara subsericea (Mart.) dubard on Biomphalaria glabrata (Say, 1818).

    Science.gov (United States)

    Faria, Robson Xavier; Rocha, Leandro Machado; Souza, Eloísa Portugal Barros Silva Soares; Almeida, Fernanda Borges; Fernandes, Caio Pinho; Santos, José Augusto Albuquerque

    2018-02-01

    Schistosomiasis is promoted for species from Schistosoma genus affecting over 200 million people worldwide. Molluscicides are an efficient method to control this disease, being able to reduce intermediate host snail Biomphalaria glabrata number. In function of resistance cases using niclosamide, natural products are promisors to discover new drugs. Manilkara subsericea is endemic to Brazilian sandbanks of Rio de Janeiro State and wide ranges of biological activities. However, there is no studies evaluating its effects as molluscicidal agent. We tested crude extract from leaves of M. subsericea molluscicidal action, as well it ethyl-acetate fraction and isolated substances against B. glabrata. M. subsericea leaves crude extract and ethyl acetate fraction induced 80±4.13% and 86.66±4.59% mortality of adult snails at concentrations of 250ppm after 96h, and their LD 50 values were 118.7±1.62 and 23.41±1.15ppm respectively. Isolated substances from M. subsericea were also considered active. Quercetin, myricetin and ursolic acid, at concentration of 100ppm (96h), were able to induce mortality levels of 100%, 80% and 53.33%, respectively. Our results suggest that M. subsericea can be considered promising as a molluscicide agent. Copyright © 2017. Published by Elsevier B.V.

  17. Pathogenesis of natural and experimental Pseudorabies virus infections in dogs.

    Science.gov (United States)

    Zhang, Letian; Zhong, Cheng; Wang, Jushi; Lu, Zijie; Liu, Lei; Yang, Wanlian; Lyu, Yanli

    2015-03-18

    Since late 2011, cases of suspected canine pseudorabies have increased in north China with the outbreak of swine pseudorabies in the same area, but the pathogenesis of canine Pseudorabies virus (PRV) infections in China is poorly understood. In this study, we investigated the pathogenesis of canine pseudorabies. The pathological changes in 13 dogs that died of natural PRV infections (confirmed by pathogen detection) during 2011-2013 in Beijing were evaluated. An experimental study was also conducted in which healthy adult beagle dogs were administered PRV isolate BJ-YT by subcutaneous injection. The dog tissues were subjected to gross and microscopic examinations and immunohistochemical analysis and the dogs' serum cardiac troponin-I (cTn-I) was measured. Systemic hemorrhage and/or congestion were the most marked pathological changes in both the naturally and experimentally PRV-infected dogs. Macroscopically, the major lesions consisted of petechiae and ecchymoses in both the endocardium and epicardium, thrombi in the mitral valves, hemorrhage in the lungs and thymus, and incomplete contraction of the spleen. Microscopically, the major histopathological findings were systemic hemorrhage and congestion, nonsuppurative ganglioneuritis (in the experimentally infected dogs, unexamined in the naturally PRV-infected dogs), brainstem encephalitis (in the naturally infected dogs), necrosis or exudation in the myocardium, and lymphoid depletion in many lymphoid organs and tissues. Viral antigens were only detected in the brainstems and peripheral ganglia of the infected dogs. Serum cTn-I was significantly higher in the experimentally PRV-infected dogs with myocardial lesions than in the dogs without myocardial lesions. Based on these results, we conclude that virally induced systemic hemorrhage, peripheral nervous system pathology, and/or cardiac injury can individually or collectively cause death in PRV-infected dogs. The respiratory signs of the disease are attributed to

  18. Rapid Discrimination between Candida glabrata, Candida nivariensis, and Candida bracarensis by Use of a Singleplex PCR ▿

    OpenAIRE

    Enache-Angoulvant, A.; Guitard, J.; Grenouillet, F.; Martin, T.; Durrens, P.; Fairhead, C.; Hennequin, C.

    2011-01-01

    We report here a PCR-based assay using a single primer pair targeting the RPL31 gene that allows discrimination between Candida glabrata, Candida bracarensis, and Candida nivariensis according to the size of the generated amplicon.

  19. Time to positivity and detection of growth in anaerobic blood culture vials predict the presence of Candida glabrata in candidemia: a two-center European cohort study.

    Science.gov (United States)

    Cobos-Trigueros, Nazaret; Kaasch, Achim J; Soriano, Alex; Torres, Jorge-Luis; Vergara, Andrea; Morata, Laura; Zboromyrska, Yuliya; De La Calle, Cristina; Alejo, Izaskun; Hernández, Cristina; Cardozo, Celia; Marco, Franscesc; Del Río, Ana; Almela, Manel; Mensa, Josep; Martínez, José Antonio

    2014-08-01

    This study shows the accuracy of exclusive or earlier growth in anaerobic vials to predict Candida glabrata in a large series of candidemic patients from two European hospitals using the Bactec 9240 system. Alternatively, C. glabrata can be predicted by a time to positivity cutoff value, which should be determined for each setting. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  20. Nonhemocyte sources of selected lysosomal enzymes in Biomphalaria glabrata, B. tenagophila and B. straminea (Mollusca: Pulmonata

    Directory of Open Access Journals (Sweden)

    Gary E. Rodrick

    1981-09-01

    Full Text Available The specific activities of acid phosphatase, alkaline phosphatase, β-glucuronidase, lysozymes, glutamate-oxalacetate transaminase and glutamate-pyruvate transaminate were determined in the head-foot and digestive gland of Brazilian Biomphalaria glabrata (Touros, B. tenagophila (Caçapava and B. straminea (Monsenhor Gil. All six enzymes were detected inthe 3000g supernatant. Both cytoplasmic enzymes, glutamate-oxalacetate and glutamate-pyruvate transaminase exhibited the highest specific activities. In the case of the four hydrolytic enzymes assayed, β-glucuronidase exhibited the highest specific activity while lysozyme showed the lowest activity. All six enzymes are thought to be produced by cells within the head-foot and digestive gland of B. glabrata, B. tenagophila and B. straminea.Foram determinadas, na massa cefalopedal e na glândula digestiva de Biomphalaria glabrata de Touros (Rio Grande do Norte B. tenagophila de Cacapava (Sao Paulo e B. straminea de Monsenhor Gil (Piauí, as atividades específicas das seguintes enzimas: fosfatase acida, fosfatase alcalina, beta-glucuronidase, lisozima, transaminase glutâmico-oxalacetica e transaminase glutâmico-piruvica. As seis enzimas referidas foram detectadas no sobrenadante a 3000g. Ambas as enzimas citoplasmaticas - transaminases glutamico-oxalacetica e glutamico-piruvica - mostraram as atividades específicas mais altas. No caso das quatro enzimas hidrolíticas, a beta-glucuronidase revelou a mais alta atividade específica, enquanto a lisozima revelou a mais baixa. E admitido que todas as seis enzimas sao produzidas por celulas presentes na massa cefalopedal e na glândula digestiva das tres especies de moluscos examinadas.

  1. Evaluation of reference genes for real-time quantitative PCR studies in Candida glabrata following azole treatment

    Directory of Open Access Journals (Sweden)

    Li Qingdi

    2012-06-01

    Full Text Available Abstract Background The selection of stable and suitable reference genes for real-time quantitative PCR (RT-qPCR is a crucial prerequisite for reliable gene expression analysis under different experimental conditions. The present study aimed to identify reference genes as internal controls for gene expression studies by RT-qPCR in azole-stimulated Candida glabrata. Results The expression stability of 16 reference genes under fluconazole stress was evaluated using fold change and standard deviation computations with the hkgFinder tool. Our data revealed that the mRNA expression levels of three ribosomal RNAs (RDN5.8, RDN18, and RDN25 remained stable in response to fluconazole, while PGK1, UBC7, and UBC13 mRNAs showed only approximately 2.9-, 3.0-, and 2.5-fold induction by azole, respectively. By contrast, mRNA levels of the other 10 reference genes (ACT1, EF1α, GAPDH, PPIA, RPL2A, RPL10, RPL13A, SDHA, TUB1, and UBC4 were dramatically increased in C. glabrata following antifungal treatment, exhibiting changes ranging from 4.5- to 32.7-fold. We also assessed the expression stability of these reference genes using the 2-ΔΔCT method and three other software packages. The stability rankings of the reference genes by geNorm and the 2-ΔΔCT method were identical to those by hkgFinder, whereas the stability rankings by BestKeeper and NormFinder were notably different. We then validated the suitability of six candidate reference genes (ACT1, PGK1, RDN5.8, RDN18, UBC7, and UBC13 as internal controls for ten target genes in this system using the comparative CT method. Our validation experiments passed for all six reference genes analyzed except RDN18, where the amplification efficiency of RDN18 was different from that of the ten target genes. Finally, we demonstrated that the relative quantification of target gene expression varied according to the endogenous control used, highlighting the importance of the choice of internal controls in such

  2. Deep, multi-stage transcriptome of the schistosomiasis vector Biomphalaria glabrata provides platform for understanding molluscan disease-related pathways

    Directory of Open Access Journals (Sweden)

    Nathan J Kenny

    2016-10-01

    Full Text Available Abstract Background The gastropod mollusc Biomphalaria glabrata is well known as a vector for the tropical disease schistosomiasis, which affects nearly 200 million people worldwide. Despite intensive study, our understanding of the genetic basis of B. glabrata development, growth and disease resistance is constrained by limited genetic resources, constraints for which next-generation sequencing methods provide a ready solution. Methods Illumina sequencing and de novo assembly using the Trinity program was used to generate a high-quality transcriptomic dataset spanning the entirety of in ovo development in schistosomiasis-free B. glabrata. This was subjected to automated (KEGG, BLAST2GO and manual annotation efforts, allowing insight into the gene complements of this species in a number of contexts. Results Excellent dataset recovery was observed, with 133,084 contigs produced of mean size 2219.48 bp. 80,952 (60.8 % returned a BLASTx hit with an E value of less than 10-3, and 74,492 (55.97 % were either mapped or assigned a GO identity using the BLAST2GO program. The CEGMA set of core eukaryotic genes was found to be 99.6 % present, indicating exceptional transcriptome completeness. We were able to identify a wealth of disease-pathway related genes within our dataset, including the Wnt, apoptosis and Notch pathways. This provides an invaluable reference point for further work into molluscan development and evolution, for studying the impact of schistosomiasis in this species, and perhaps providing targets for the treatment of this widespread disease. Conclusions Here we present a deep transcriptome of an embryonic sample of schistosomiasis-free B. glabrata, presenting a comprehensive dataset for comparison to disease-affected specimens and from which conclusions can be drawn about the genetics of this widespread medical model. Furthermore, the dataset provided by this sequencing provides a useful reference point for comparison to other mollusc

  3. Primary peak and chronic malaria infection levels are correlated in experimentally infected great reed warblers.

    Science.gov (United States)

    Asghar, Muhammad; Westerdahl, Helena; Zehtindjiev, Pavel; Ilieva, Mihaela; Hasselquist, Dennis; Bensch, Staffan

    2012-09-01

    Malaria parasites often manage to maintain an infection for several months or years in their vertebrate hosts. In humans, rodents and birds, most of the fitness costs associated with malaria infections are in the short initial primary (high parasitaemia) phase of the infection, whereas the chronic phase (low parasitaemia) is more benign to the host. In wild birds, malaria parasites have mainly been studied during the chronic phase of the infection. This is because the initial primary phase of infection is short in duration and infected birds with severe disease symptoms tend to hide in sheltered places and are thus rarely caught and sampled. We therefore wanted to investigate the relationship between the parasitaemia during the primary and chronic phases of the infection using an experimental infection approach. We found a significant positive correlation between parasitaemia in the primary peak and the subsequent chronic phase of infection when we experimentally infected great reed warblers (Acrocephalus arundinaceus) with Plasmodium ashfordi. The reason for this association remains to be understood, but might arise from individual variation in exoerythrocytic parasite reservoirs in hosts, parasite antigenic diversity and/or host genetics. Our results suggest that the chronic phase parasitaemia can be used to qualitatively infer the parasitaemia of the preceding and more severe primary phase, which is a very important finding for studies of avian malaria in wild populations.

  4. Potassium Uptake Mediated by Trk1 Is Crucial for Candida glabrata Growth and Fitness

    Czech Academy of Sciences Publication Activity Database

    Llopis-Torregrosa, Vincent; Hušeková, Barbora; Sychrová, Hana

    2016-01-01

    Roč. 11, č. 4 (2016), e0153374 E-ISSN 1932-6203 R&D Projects: GA ČR(CZ) GAP302/12/1151; GA ČR(CZ) GA16-03398S EU Projects: European Commission(XE) 606786 - ImResFun Institutional support: RVO:67985823 Keywords : Candida glabrata * potassium uptake * Trk1 protein Subject RIV: EE - Microbiology, Virology Impact factor: 2.806, year: 2016

  5. Primary infection protects pigs against re-infection with Lawsonia intracellularis in experimental challenge studies

    DEFF Research Database (Denmark)

    Riber, Ulla; Hvass, Henriette Cordes; Boutrup, Torsten Snogdal

    2011-01-01

    In two separate trials previous termpigsnext term were experimentally infected with previous termLawsonia intracellularisnext term at 5–6 weeks of age followed by antibiotic treatment and resolution of the previous termprimary infection and then renext term-inoculated at 12–13 weeks of age. A tre...

  6. Oxidative stress in rats experimentally infected by Sporothrix schenckii.

    Science.gov (United States)

    Castro, Verônica S P; Da Silva, Aleksandro S; Thomé, Gustavo R; Wolkmer, Patrícia; Castro, Jorge L C; Costa, Márcio M; Graça, Dominguita L; Oliveira, Daniele C; Alves, Sydney H; Schetinger, Maria R C; Lopes, Sonia T A; Stefani, Lenita M; Azevedo, Maria I; Baldissera, Matheus D; Andrade, Cinthia M

    2017-06-01

    The aim of this study was to evaluate whether oxidative stress occurs in rats experimentally infected by Sporothrix schenckii, and its possible effect on disease pathogenesis. Thirty rats were divided into two groups: the group A (uninfected, n = 18) and the group B (infected by S. schenckii, n=21). Blood samples were collected on days 15, 30 and 40 post-infection (PI). At each sampling time, six rats of the group A, and seven of the group B were bled. TBARS (thiobarbituric acid reactive substances) levels in serum samples were measured to evaluate lipid peroxidation. In addition, catalase (CAT) and superoxide dismutase (SOD) activities, known as biomarkers of antioxidants levels, were verified in whole blood. Seric pro-inflammatory cytokine levels were measured (IFN-γ, TNF-α, and IL-6), which showed that these inflammatory mediators were at higher levels in the infected rats (P sporotrichosis showed significantly higher (p sporotrichosis is a likely mechanism for redox imbalance, and consequently cause the oxidative stress in experimentally infected rats. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Influence of gamma radiation on the levels of polyphenols and lethality of ethanol extracts of Anacardium occidentale Linn., against Biomphalaria glabrata

    International Nuclear Information System (INIS)

    Santos, G.H.F.; Silva, E.B.; Melo, A.M.M.A.; Lima, C.S.A; Amorim, E.L.C.; Peixoto Sobrinho, T.J.S.

    2013-01-01

    Plant materials rich in phenolic compounds, such as Anacardium occidentale Linn., Have been used as alternatives to synthetic pesticides in Biomphalaria glabrata control programs, intermediate host of Schistosoma mansoni. Studies show that ionizing radiation can influence the content of phenolic compounds and thus their biological actions. The aim of this study was to evaluate the influence of gamma radiation of 60 Co in polyphenol composition of hydroalcoholic extracts of bark and leaves of A. occidentale and evaluate the toxicity of these extracts to embryos and adults of B. glabrata. To achieve this goal this, the extracts were irradiated at 10 kGy, the controls being maintained from 0 kGy and positive (CaCO 3 ) and negative (H 2 O). We quantified the total phenols by the Folin-Ciocalteau and tannins by precipitation of casein. Extracts were used at a concentration of 100 mg/L. The results showed that the radiation caused the changes to the leaves, the percentage of polyphenols and tannins, and the percentage of lethality in embryos and adults Biomphalaria glabrata, these percentages being: 13 ± 5 (0 kGy) and 27 ± 2.5 (10 kGy), and 36.67 ± 5.77 (0 kGy), and 56.67 ± 5.77 (10 kGy), respectively. Gamma radiation caused significant changes in the levels of polyphenols in the extracts of leaves of Anacardium ocidentale Linn., translated by the increased toxicity of this extract against embryos and adults of Biomphalaria glabrata. This indicates that gamma radiation can be used as an agent potentiating the toxicity of plant extracts on the alternate use of these materials as molluscicides. (author)

  8. Molluscicidal effect of Euphorbia umbellata (Pax Bruyns latex on Biomphalaria glabrata, Schistosoma mansoni host snail

    Directory of Open Access Journals (Sweden)

    Luciana Patrícia Lima Alves Pereira

    2017-12-01

    Full Text Available ABSTRACT Euphorbia umbellata (Pax Bruyns is an easily cultivated shrub, with occurrence in the tropical regions of the American and African continents. Chemical studies have revealed that the latex of this plant is rich in terpene compounds, which are highly toxic to snails Biomphalaria glabrata (Basommatophora: Planorbidae. The aim of this study was to evaluate the chemical composition and molluscicidal activity of the latex produced by E. umbellata, as well as the safety of its application in aquatic environments. The concentration of latex that killed 90% of the exposed snails after 24 h exposure (LC90 was 3.69 mg/L. Toxicity bioassays using Danio rerio (zebrafish revealed that these animals were less susceptible to latex than planorbids. However, it is important to perform other toxicity tests to ensure the feasibility of using latex to control populations of mollusks that contribute to schistosomiasis transmission. A phytochemical screening performed with the E. umbellata latex identified the triterpenoid and coumarin class. Further studies are warranted to isolate, identify, and test the active compounds of E. umbellata latex in B. glabrata.

  9. Enhanced oxidative killing of azole-resistant Candida glabrata strains with ERG11 deletion.

    Science.gov (United States)

    Kan, V L; Geber, A; Bennett, J E

    1996-01-01

    The susceptibility of genetically defined Candida glabrata strains to killing by H2O2 and neutrophils was assessed. Fluconazole-susceptible L5L and L5D strains demonstrated survival rates higher than those of two fluconazole-resistant strains lacking the ERG11 gene coding for 14 alpha-demethylase. Fluconazole resistance can occur by mechanisms which increase fungal susceptibility to oxidative killing by H2O2 and neutrophils. PMID:8807069

  10. Changes in energetic metabolism of Biomphalaria glabrata (Mollusca, Planorbidae in response to exogenous calcium

    Directory of Open Access Journals (Sweden)

    L. D. Silva

    Full Text Available Abstract Calcium is considered an essential element for the metabolism of aquatic snail Biomphalaria glabrata (Say, 1818, intermediate host of Schistosoma mansoni Sambon, 1907 in Brazil, and represents a limiting factor to its distribution and adaptation to the environment. This study investigated the effect of different concentrations of exogenous CaCO3 on the energetic metabolism of B. glabrata for better understanding the physiological interference of chemical elements dissolved in the environment with the physiology of this species. Sixty-day-old snails were distributed into six groups, five exposed to different concentrations of CaCO3 (20, 40, 60, 80 and 100 mg/L and a control group. The exposure to CaCO3 was assessed over time, with analysis of 15 snails of each group in the following intervals: 1, 14, 21 or 30 days for hemolymph extraction. Concentrations of calcium and glucose in the hemolymph were determined by commercial kits, and organic acids were extracted using an ion exchange column and analyzed by high-performance liquid chromatography. Concentration of calcium in the hemolymph showed no significant difference (p>0.05 from the control group and between the concentrations tested. Concentration of glucose decreased (p<0.05 in the treatments of exposure to 20 and 40 mg/L and increased when exposed to 80 and 100 mg/L CaCO3 compared to control and to other concentrations tested over 30 days. The organic acids pyruvate, oxaloacetate, citrate, succinate, fumarate, beta-hydroxybutyrate and lactate presented increased concentrations, while propionate and acetoacetate, decreased concentrations, when exposed to CaCO3 compared to control. Considering the influence of different periods of exposure to CaCO3, on the 14th day, there were stronger alterations in the metabolism of B. glabrata. In conclusion, exposure to CaCO3 reduced the concentration of glucose, which is metabolized into pyruvate, the final product of glycolysis, and also

  11. SNF3 as high affinity glucose sensor and its function in supporting the viability of Candida glabrata under glucose-limited environment

    Directory of Open Access Journals (Sweden)

    Tzu Shan eNg

    2015-12-01

    Full Text Available Candida glabrata is an emerging human fungal pathogen that has efficacious nutrient sensing and responsiveness ability. It can be seen through its ability to thrive in diverse range of nutrient limited-human anatomical sites. Therefore, nutrient sensing particularly glucose sensing is thought to be crucial in contributing to the development and fitness of the pathogen. This study aimed to elucidate the role of SNF3 (Sucrose Non Fermenting 3 as a glucose sensor and its possible role in contributing to the fitness and survivability of C. glabrata in glucose-limited environment. The SNF3 knockout strain was constructed and subjected to different glucose concentrations to evaluate its growth, biofilm formation, amphotericin B susceptibility, ex vivo survivability and effects on the transcriptional profiling of the sugar receptor repressor (SRR pathway-related genes. The SNF3Δ strain showed a retarded growth in low glucose environments (0.01% and 0.1% in both fermentation and respiration-preferred conditions but grew well in high glucose concentration environments (1% and 2%. It was also found to be more susceptible to amphotericin B in low glucose environment (0.1% and macrophage engulfment but showed no difference in the biofilm formation capability. The deletion of SNF3 also resulted in the down-regulation of about half of hexose transporters genes (4 out of 9. Overall, the deletion of SNF3 causes significant reduction in the ability of C. glabrata to sense limited surrounding glucose and consequently disrupts its competency to transport and perform the uptake of this critical nutrient. This study highlighted the role of SNF3 as a high affinity glucose sensor and its role in aiding the survivability of C. glabrata particularly in glucose limited environment.

  12. Detection of toxoplasma gondii antigens in sera from experimentally infected mice

    International Nuclear Information System (INIS)

    Shojaee, S.; Keshavarz, H.; Rezaian, M.; Mohebali, M.

    2007-01-01

    Detection of Toxoplasma antigen in serum of mice by Immunoblotting. strain. IgG isolated from rabbits that were immunized with T. gondii Immunoblotting was performed to detect T. gondii antigens in sera of mice. Serum samples from mice experimentally infected with T. gondii RH strain. The value of Immunoblotting in diagnosis of toxoplasmosis in acute stage of infection. The antigen bands detected in serum sample of mice were experimentally infected with T. gondii tachyzoite in immunoblotting. Six bands demonstrated on seventh post infection day six bands were identified. Similarly on sixth day four bands, on day five three bands and on fourth post infection day two bands were identified. No band was detected in control group sera. Immunoblotting is a sensitive method for diagnosis of acute stage of toxoplasmosis. (author)

  13. Infectivity and temperature tolerance of non-encapsulating Trichinella zimbabwensis in experimentally infected red foxes (Vulpes vulpes)

    DEFF Research Database (Denmark)

    Hurníková, Z.; Dubinský, P.; Mukaratirwa, S.

    2004-01-01

    The non-encapsulating Trichinella zimbabwensis was evaluated for infectivity in red foxes (Vulpes vulpes), the larval distribution and cold tolerance in fox muscle tissue. Six red foxes were experimentally infected with T. zimbabwensis larvae. Five weeks after inoculation, muscle larvae were...... recovered from 9 different muscle types using artificial digestion method. The establishment of infection in all infected red foxes demonstrated the ability of T. zimbabwensis to complete its life cycle in a carnivore mammal host. The larvae recovered from fox muscle tissue were infective to mice, they have...

  14. Experimental porcine cysticercosis using infected beetles with Taenia solium eggs.

    Science.gov (United States)

    Gomez-Puerta, Luis A; Garcia, Hector H; Gonzalez, Armando E

    2018-07-01

    Beetles are intermediate hosts for human and animal parasites, and several beetle species have been shown to carry Taenia eggs. An experimental porcine cysticercosis infection model was developed using beetles (Ammophorus rubripes) infected with Taenia solium eggs and then using these beetles for oral pig challenge. A total of 18 three months-old Landrace pigs were divided in four groups. Pigs from groups 1, 2, and 3 (n = 6 pigs per group) were challenged with one, three, and six beetles infected with T. solium eggs, containing approximately 52, 156 or 312 eggs respectively. Pigs were necropsied 12 weeks after infection to assess the presence of T. solium metacestode. Porcine cysticercosis by T. solium was produced in 17 out of 18 pigs (94.4%) challenged with infected beetles, all infected pigs had viable cysts. Only one pig from group 1 was negative to the presence of cysts. The median number of metacestodes per pig in groups 1, 2, and 3 were 2 (range 0-71), 26 (range 5-33) and 40 cysts (range 4-111), respectively. Experimental porcine cysticercosis infection is consistently obtained using beetles as mechanical vectors for T. solium eggs. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. FKS2 Mutations Associated with Decreased Echinocandin Susceptibility of Candida glabrata following Anidulafungin Therapy

    OpenAIRE

    Costa-de-Oliveira, Sofia; Miranda, Isabel Marcos; Silva, Raquel M.; Pinto e Silva, Ana; Rocha, Rita; Amorim, Antonio; Rodrigues, Acacio Goncalves; Pina-Vaz, Cidalia

    2011-01-01

    This is the first case report of Candida glabrata-disseminated candidiasis describing the acquisition of echinocandin resistance following anidulafungin treatment. The initial isolates recovered were susceptible to echinocandins. However, during 27 days of anidulafungin treatment, two resistant strains were isolated (from the blood and peritoneal fluid). The resistant peritoneal fluid isolate exhibited a Ser663Pro mutation in position 1987 of FKS2 HS1 (hot spot 1), whereas the resistant blood...

  16. Humoral immune response and spreading of Encephalitozoon cuniculi infection in experimentally infected ponies

    Czech Academy of Sciences Publication Activity Database

    Wagnerová, Pavla; Sak, Bohumil; Květoňová, Dana; Maršálek, M.; Langerová, I.; Kváč, Martin

    2013-01-01

    Roč. 197, 1-2 (2013), s. 1-6 ISSN 0304-4017 Institutional support: RVO:60077344 Keywords : Ponies * Encephalitozoon cuniculi genotype II * PCR * Antibodies * Experimental infection Subject RIV: EC - Immunology Impact factor: 2.545, year: 2013

  17. Duration of pairing and use of allosperm in Biomphalaria glabrata (Gastropoda: Planorbidae Duração do acasalamento e uso do aloesperma em Biomphalaria glabrata (Gastropode: Planorbidae

    Directory of Open Access Journals (Sweden)

    Marc Vianey-Liaud

    1989-03-01

    Full Text Available Virgin homozygous black pigmented and albino Biomphalaria glabrata are paired during a period varying from 1 to 20 days. The rate of cross-fertilized parents is statistically similar for the various lengths of pairing. As a whole, nearly 80% of the albino snails produce a pigmented progeny. This production begins as soon as the snails are mated and continues after their separation. To measure the actual use of the allosperm, its use during the postmating period must be added to the length of mating. So, it appears that the real use of the allosperm is statistically constant (mean slightly inferior to 8 weeks and not related to the length of the previous pairing.Espécimes virgens de Biomphalaria glabrata (um melâncio e um albino são acasalados durante um período variável de 1 a 20 dias. A proporção de espécimes parentais albinos fecundados por cruzamento é semelhante estatisticamente para os diversos períodos de acasalamento. Em conjunto, cerca de 80% dos albinos produziram descendência pigmentada. Essa produção começa assim que os moluscos são acasalados e continua depois que são separados. Para medir a utilização real do aloesperma, sua utilização durante o período de pós-acasalamento. Parece, assim, que a utilização real do aloesperma é estatisticamente constante (média ligeiramente inferior a oito semanas e não relacionada a duração do acasalamento anterior.

  18. Effects of Experimental Sarcocystis neurona-Induced Infection on Immunity in an Equine Model

    Directory of Open Access Journals (Sweden)

    S. Rochelle Lewis

    2014-01-01

    Full Text Available Sarcocystis neurona is the most common cause of Equine Protozoal Myeloencephalitis (EPM, affecting 0.5–1% horses in the United States during their lifetimes. The objective of this study was to evaluate the equine immune responses in an experimentally induced Sarcocystis neurona infection model. Neurologic parameters were recorded prior to and throughout the 70-day study by blinded investigators. Recombinant SnSAG1 ELISA for serum and CSF were used to confirm and track disease progression. All experimentally infected horses displayed neurologic signs after infection. Neutrophils, monocytes, and lymphocytes from infected horses displayed significantly delayed apoptosis at some time points. Cell proliferation was significantly increased in S. neurona-infected horses when stimulated nonspecifically with PMA/I but significantly decreased when stimulated with S. neurona compared to controls. Collectively, our results suggest that horses experimentally infected with S. neurona manifest impaired antigen specific response to S. neurona, which could be a function of altered antigen presentation, lack of antigen recognition, or both.

  19. Effects of Experimental Sarcocystis neurona-Induced Infection on Immunity in an Equine Model.

    Science.gov (United States)

    Lewis, S Rochelle; Ellison, Siobhan P; Dascanio, John J; Lindsay, David S; Gogal, Robert M; Werre, Stephen R; Surendran, Naveen; Breen, Meghan E; Heid, Bettina M; Andrews, Frank M; Buechner-Maxwell, Virginia A; Witonsky, Sharon G

    2014-01-01

    Sarcocystis neurona is the most common cause of Equine Protozoal Myeloencephalitis (EPM), affecting 0.5-1% horses in the United States during their lifetimes. The objective of this study was to evaluate the equine immune responses in an experimentally induced Sarcocystis neurona infection model. Neurologic parameters were recorded prior to and throughout the 70-day study by blinded investigators. Recombinant SnSAG1 ELISA for serum and CSF were used to confirm and track disease progression. All experimentally infected horses displayed neurologic signs after infection. Neutrophils, monocytes, and lymphocytes from infected horses displayed significantly delayed apoptosis at some time points. Cell proliferation was significantly increased in S. neurona-infected horses when stimulated nonspecifically with PMA/I but significantly decreased when stimulated with S. neurona compared to controls. Collectively, our results suggest that horses experimentally infected with S. neurona manifest impaired antigen specific response to S. neurona, which could be a function of altered antigen presentation, lack of antigen recognition, or both.

  20. [Experimental infection of the gerbil (Meriones unguiculatus) by Colombian isolates of Giardia duodenalis].

    Science.gov (United States)

    Arévalo, Adriana; Duque, Sofía; Nicholls, Rubén Santiago

    2005-09-01

    Natural and experimental Giardia infections have been reported from bovines, equines, goats, canines, felines and rodents such as mice, rats and gerbils. The latter have provided successful animal models for Giardia duodenalis and Giardia muris experimental infections. The gerbil model was used to establish the pattern of infection of Colombian Giardia human isolates. Giardia cysts were obtained from stool specimens of symptomatic giardiasis patients by means of sucrose-percoll gradients. Animal inoculation was performed by gastric intubation and injection with 5 x 10(3) Giardia cysts. The course of infection was established by counting cysts every day and trophozoites weekly throughout a period of 30 days. The pattern of cyst excretion was found to be intermittent. Cysts were released during the second and third weeks of infection but not during the first or fourth weeks. The mean minimal number of cysts released per 2-hr collection period was 79 and the mean maximum number was 17,943. Colonization of the small intestine by trophozoites was observed with a mean number ranging from 15,000 to 6,577,778 trophozoites/ml. Gerbils inoculated with G. duodenalis isolates obtained from geographical areas outside Colombia resolved the infection between 86 and 114 days after infection, whereas gerbils infected with Colombian G. duodenalis isolates resolved the infection at 30 days. The gerbil proved to be a good animal model for experimental infection with Colombian isolates of G. duodenalis. Experimental Giardia infection of gerbils permit a sufficient yield of cysts and trophozoites to be used as antigens for the immunization of other animals and to obtain Giardia antibodies that could be used for Giardia antigen detection assays in stool specimens.

  1. Antifungal activity of caspofungin in experimental infective endocarditis caused by Candida albicans.

    Science.gov (United States)

    Victorio, Gerardo Becerra; Bourdon, Lorena Michele Brennan; Benavides, Leonel García; Huerta-Olvera, Selene G; Plascencia, Arturo; Villanueva, José; Martinez-Lopez, Erika; Hernández-Cañaveral, Iván Isidro

    2017-05-01

    Infective endocarditis is a disease characterised by heart valve lesions, which exhibit extracellular matrix proteins that act as a physical barrier to prevent the passage of antimicrobial agents. The genus Candida has acquired clinical importance given that it is increasingly being isolated from cases of nosocomial infections. To evaluate the activity of caspofungin compared to that of liposomal amphotericin B against Candida albicans in experimental infective endocarditis. Wistar rats underwent surgical intervention and infection with strains of C. albicans to develop infective endocarditis. Three groups were formed: the first group was treated with caspofungin, the second with liposomal amphotericin B, and the third received a placebo. In vitro sensitivity was first determined to further evaluate the effect of these treatments on a rat experimental model of endocarditis by semiquantitative culture of fibrinous vegetations and histological analysis. Our semiquantitative culture of growing vegetation showed massive C. albicans colonisation in rats without treatment, whereas rats treated with caspofungin showed significantly reduced colonisation, which was similar to the results obtained with liposomal amphotericin B. The antifungal activity of caspofungin is similar to that of liposomal amphotericin B in an experimental model of infective endocarditis caused by C. albicans.

  2. Fluconazole affects the alkali-metal-cation homeostasis and susceptibility to cationic toxic compounds of Candida glabrata

    Czech Academy of Sciences Publication Activity Database

    Elicharová, Hana; Sychrová, Hana

    2014-01-01

    Roč. 160, č. 8 (2014), s. 1705-1713 ISSN 1350-0872 R&D Projects: GA ČR GAP302/12/1151; GA MŠk(CZ) ED1.1.00/02.0109 Institutional research plan: CEZ:AV0Z50110509 Institutional support: RVO:67985823 Keywords : Candida glabrata * fluconazole * membrane potential Subject RIV: EE - Microbiology, Virology Impact factor: 2.557, year: 2014

  3. Effects of CO-60 gamma radiation on the embryonary development of Biomphalaria Glabrata (Say, 1818)

    International Nuclear Information System (INIS)

    Okazaki, K.

    1988-01-01

    Some aspects of the effects of the ionizing radiation on the embryo and on the genetical material of Biomphalaria glabrata (Mollusca: Gastropoda) are presented. The embryos weresubmitted at various stages of development to doses of 5,10,15,20 and 25 Gy of Co-60 gamma radiation. As a criteia of evaluation of the embryos radiosensitivity, four biological parameters were used: mortality, malformation, hatching and chromossomal aberrations. (M.A.C.) [pt

  4. [Infections of the oral mucosa II. Bacterial, mycotic and viral infections].

    Science.gov (United States)

    Reichart, P A

    1999-11-01

    Non-specific infections of the oral mucosa are rare; however, they may present during HIV infection in the form of gingivo-periodontal lesions. In some of these Candida albicans may play a role in the pathogenesis. Sexually transmitted bacterial infections such as gonorrhoea and syphilis are frequently associated with HIV infection. Since penicillin resistance is frequent in gonorrhoea, the cephalosporines are mainly used for treatment. Syphilis increases the risk for transmission of HIV. Lues maligna with oral manifestations has been described. For this, penicillin G is the therapy of choice. Tuberculosis, characterized by multitherapy resistance, is associated with HIV infections world-wide; oral manifestations are rare. Oral candidiasis during HIV infection is often characterized by therapy resistance against fluconazole and a shift in species, with Candida glabrata and Candida krusei as the emerging species. The azoles are still the mainstay of therapy, particularly fluconazole. Herpes simplex (HSV) infections run an atypical course during HIV disease; resistance against acyclovir is a clinical problem. The association of HSV infection with erythema exudativum multiforme has been clearly shown. Oral hairy leukoplakia caused by Epstein Barr virus is a characteristic infection during immunosuppression. Cytomegalovirus infection is also observed in immunodeficient patients. Cases of ganciclovir resistance have been described. Human herpes virus 8 (HHV 8) is associated with Kaposi's sarcoma. Therapeutic trials have focussed on the inhibition of HHV 8 replication. Over 100 different genotypes of human papillomaviruses are known; some can cause infections of the oral mucosa. Characteristic lesions caused by different HPV genotypes are verruca vulgaris, condyloma acuminatum and focal epithelial hyperplasia.

  5. Experimental reproduction of an Enterococcus cecorum infection in Pekin ducks.

    Science.gov (United States)

    Jung, Arne; Metzner, Martin; Köhler-Repp, Dagmar; Rautenschlein, Silke

    2013-12-01

    Enterococcus cecorum (EC) was thus far only known as a pathogen for broilers and broiler breeders. Recently there was evidence of EC field outbreaks in Pekin duck flocks in Germany. In this study we experimentally reproduced an EC infection in Pekin ducks. At 12 days post hatch, groups of Pekin ducks were infected orally, via the thoracic air sac or intravenously with 1.5 × 10(9) colony-forming units (CFU) of EC per bird or via the air sac with 8.5 × 10(5) or 8.5 × 10(7) CFU per bird. Ducks of the intravenously infected group showed 100% mortality after 2 days post infection. The air sac inoculated high-dose group exhibited a mortality rate of 67%. Birds that were infected with 8.5 × 10(5) and 8.5 × 10(7) CFU showed 6.7% mortality after 7 days post infection. Dead birds displayed pneumonia, airsacculitis, pericarditis and splenitis and EC was re-isolated from these organs. Surviving birds of all groups apart from the orally infected ducks demonstrated clinical signs such as huddling, reduced mobility and diarrhoea. Furthermore, they showed gross pathological lesions including airsacculitis and splenitis and lower bodyweights than the control group at necropsy on days 7, 14 and 21 post infection. The present study clearly confirms that EC is pathogenic for Pekin ducks after experimental infection via the intravenous route or the respiratory tract. EC therefore has to be considered as an emerging avian pathogen not only in broilers but also in Pekin ducks.

  6. BgTEP: An Antiprotease Involved in Innate Immune Sensing in Biomphalaria glabrata

    Directory of Open Access Journals (Sweden)

    Anaïs Portet

    2018-05-01

    Full Text Available Insect thioester-containing protein (iTEP is the most recently defined group among the thioester-containing protein (TEP superfamily. TEPs are key components of the immune system, and iTEPs from flies and mosquitoes were shown to be major immune weapons. Initially characterized from insects, TEP genes homologous to iTEP were further described from several other invertebrates including arthropods, cniderians, and mollusks albeit with few functional characterizations. In the freshwater snail Biomphalaria glabrata, a vector of the schistosomiasis disease, the presence of a TEP protein (BgTEP was previously described in a well-defined immune complex involving snail lectins (fibrinogen-related proteins and schistosome parasite mucins (SmPoMuc. To investigate the potential role of BgTEP in the immune response of the snail, we first characterized its genomic organization and its predicted protein structure. A phylogenetic analysis clustered BgTEP in a well-conserved subgroup of mollusk TEP. We then investigated the BgTEP expression profile in different snail tissues and followed immune challenges using different kinds of intruders during infection kinetics. Results revealed that BgTEP is particularly expressed in hemocytes, the immune-specialized cells in invertebrates, and is secreted into the hemolymph. Transcriptomic results further evidenced an intruder-dependent differential expression pattern of BgTEP, while interactome experiments showed that BgTEP is capable of binding to the surface of different microbes and parasite either in its full length form or in processed forms. An immunolocalization approach during snail infection by the Schistosoma mansoni parasite revealed that BgTEP is solely expressed by a subtype of hemocytes, the blast-like cells. This hemocyte subtype is present in the hemocytic capsule surrounding the parasite, suggesting a potential role in the parasite clearance by encapsulation. Through this work, we report the first

  7. Histopathological study of experimental and natural infections by Trypanosoma cruzi in Didelphis marsupialis

    Directory of Open Access Journals (Sweden)

    João Carlos Araujo Carreira

    1996-10-01

    Full Text Available Didelphis marsupialis, the most important sylvatic reservoir of Trypanosoma cruzi, can also maintain in their anal scent glands the multiplicative forms only described in the intestinal tract of triatomine bugs. A study of 21 experimentally and 10 naturally infected opossums with T. cruzi was undertaken in order to establish the histopathological pattern under different conditions. Our results showed that the inflammation was predominantly lymphomacrophagic and more severe in the naturally infected animals but never as intense as those described in Chagas' disease or in other animal models. The parasitism in both groups was always mild with very scarce amastigote nests in the tissues. In the experimentally infected animals, the inflammation was directly related to the presence of amastigotes nests. Four 24 days-old animals, still in embryonic stage, showed multiple amastigotes nests and moderate inflammatory reactions, but even so they survived longer and presented less severe lesions than experimentally infected adult mice. Parasites were found in smooth, cardiac and/or predominantly striated muscles, as well as in nerve cells. Differing from the experimentally infected opossums parasitism in the naturally infected animals predominated in the heart, esophagus and stomach. Parasitism of the scent glands did not affect the histopathological pattern observed in extraglandular tissues.

  8. Biomphalaria species distribution and its effect on human Schistosoma mansoni infection in an irrigated area used for rice cultivation in northeast Brazil

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    Delmany Moitinho Barboza

    2012-09-01

    Full Text Available The role of irrigated areas for the spread of schistosomiasis is of worldwide concern. The aim of the present study was to investigate the spatial distribution of the intermediate snail host Biomphalaria in an area highly endemic for schistosomiasis due to Schistosoma mansoni, evaluating the relationship between irrigation and types of natural water sources on one hand, and the influence of place and time of water exposure on the intensity of human infection on the other. A geographical information system (GIS was used to map the distribution of the intermediate snail hosts in Ilha das Flores, Sergipe, Brazil, combined with a clinical/epidemiological survey. We observed a direct correlation between the intensity of human infection with S. mansoni and irrigation projects. Malacological studies to identify snail species and infection rates showed that B. glabrata is the main species responsible for human schistosomiasis in the municipality, but that B. straminea also plays a role. Our results provide evidence for a competitive selection between the two snail species in rice fields with a predominance of B. glabrata in irrigation systems and B. straminea in natural water sources.

  9. Cardiac complication after experimental human malaria infection: a case report

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    Druilhe Pierre

    2009-12-01

    Full Text Available Abstract A 20 year-old healthy female volunteer participated in a clinical Phase I and IIa safety and efficacy trial with candidate malaria vaccine PfLSA-3-rec adjuvanted with aluminium hydroxide. Eleven weeks after the third and last immunization she was experimentally infected by bites of Plasmodium falciparum-infected mosquitoes. When the thick blood smear became positive, at day 11, she was treated with artemether/lumefantrine according to protocol. On day 16 post-infection i.e. two days after completion of treatment, she woke up with retrosternal chest pain. She was diagnosed as acute coronary syndrome and treated accordingly. She recovered quickly and her follow-up was uneventful. Whether the event was related to the study procedures such as the preceding vaccinations, malaria infection or antimalarial drugs remains elusive. However, the relation in time with the experimental malaria infection and apparent absence of an underlying condition makes the infection the most probable trigger. This is in striking contrast, however, with the millions of malaria cases each year and the fact that such complication has never been reported in the literature. The rare occurrence of cardiac events with any of the preceding study procedures may even support a coincidental finding. Apart from acute coronary syndrome, myocarditis can be considered as a final diagnosis, but the true nature and patho-physiological explanation of the event remain unclear.

  10. Evaluation of a linear spectral mixture model and vegetation indices (NDVI and EVI) in a study of schistosomiasis mansoni and Biomphalaria glabrata distribution in the state of Minas Gerais, Brazil.

    Science.gov (United States)

    Guimarães, Ricardo J P S; Freitas, Corina C; Dutra, Luciano V; Scholte, Ronaldo G C; Amaral, Ronaldo S; Drummond, Sandra C; Shimabukuro, Yosio E; Oliveira, Guilherme C; Carvalho, Omar S

    2010-07-01

    This paper analyses the associations between Normalized Difference Vegetation Index (NDVI) and Enhanced Vegetation Index (EVI) on the prevalence of schistosomiasis and the presence of Biomphalaria glabrata in the state of Minas Gerais (MG), Brazil. Additionally, vegetation, soil and shade fraction images were created using a Linear Spectral Mixture Model (LSMM) from the blue, red and infrared channels of the Moderate Resolution Imaging Spectroradiometer spaceborne sensor and the relationship between these images and the prevalence of schistosomiasis and the presence of B. glabrata was analysed. First, we found a high correlation between the vegetation fraction image and EVI and second, a high correlation between soil fraction image and NDVI. The results also indicate that there was a positive correlation between prevalence and the vegetation fraction image (July 2002), a negative correlation between prevalence and the soil fraction image (July 2002) and a positive correlation between B. glabrata and the shade fraction image (July 2002). This paper demonstrates that the LSMM variables can be used as a substitute for the standard vegetation indices (EVI and NDVI) to determine and delimit risk areas for B. glabrata and schistosomiasis in MG, which can be used to improve the allocation of resources for disease control.

  11. HoBi-like pestivirus experimental infection in pregnant ewes: Reproductive disorders and generation of persistently infected lambs.

    Science.gov (United States)

    Decaro, Nicola; Losurdo, Michele; Larocca, Vittorio; Lucente, Maria Stella; Mari, Viviana; Varello, Katia; Patruno, Giovanni; Camero, Michele; Sciarra, Marina; Occhiogrosso, Leonardo; Tempesta, Maria; Iulini, Barbara; Buonavoglia, Canio

    2015-08-05

    In order to evaluate sheep as experimental model to test the efficacy of HoBi-like pestivirus vaccines for cattle, 10 sheep at different stages of pregnancy (30 or 50 days) were experimentally infected with the Italian prototype isolate Italy-1/10-1. Irrespective of the stage of pregnancy, virus inoculation resulted in reproductive failures, consisting of abortion, stillbirths or birth of weak or persistently infected (PI) lambs. Aborted fetuses, stillborn and dead lambs displayed extensive histopathological changes, consisting of hemorrhages, congestion and mononuclear infiltration in major organs. Pestiviral antigens were detected by immunohistochemistry in most tissues with remarkable signals in lungs and kidneys. PI lambs were constantly viremic, shed the virus through the nasal secretions and feces and, in all cases but one, did not have detectable HoBi-like pestivirus antibodies before the assumption of colostrum. The single seropositive infected lamb showed low-titer viremia and viral shedding that ceased only several weeks after the 3-month observation period. The study proves that sheep are susceptible to the reproduction failures caused by HoBi-like pestivirus infection and can serve as a suitable model for the evaluation of the fetal protection induced by homologous experimental vaccines. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Morphology and anatomy of leaf miners in two species of Commelinaceae (Commelina diffusa Burm. f. and Floscopa glabrata (Kunth Hassk Morfologia e anatomia de minas foliares em duas espécies de Commelinaceae Commelina diffusa Burm. f. e Floscopa glabrata (Kunth Hassk

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    Paula Maria Elb

    2010-03-01

    Full Text Available In specialized literature, reports on anatomy of miners in host plants are few in number. These agents trigger excavations, or paths, by consumption of plant inner tissues by larvae of several insects. The aim of this work was to investigate leaf miner occurrence in Commelina diffusa (a cosmopolitan plant and Floscopa glabrata (an amphibious plant using anatomical techniques. The place where the plants were collected is subjected to seasonal floods, consequently both the species were exposed to the same weather conditions and seasonal floods. This study showed that members of Agromyzidae and Chironomidae families, which are Diptera endophytophagous larvae types, were responsible for the tunnels. Moreover, in Commelina diffusa Agromyzidae larvae were found, while in Floscopa glabrata three Chironomidae cephalic exuviae were found. The miners, as can be seen from anatomical studies, used only mesophyll parenchyma tissues for feeding, causing the formation of linear mines. In addition, in both the species, the epidermis and the medium-sized vascular units were kept intact, showing no structural modification, such as neoformation of tissues.Existem poucos relatos na literatura sobre anatomia de plantas parasitadas por agentes minadores, os quais promovem escavações ou caminhos através do consumo dos tecidos internos das plantas por larvas de diversos insetos. A proposta deste trabalho foi analisar anatomicamente a ocorrência de minas foliares em Commelina diffusa (planta cosmopolita e Floscopa glabrata (planta anfíbia causadas por espécies de larvas endofitófagas de dípteros, pertencentes a duas famílias: Agromyzidae e Chironomidae. O local onde as plantas foram coletas está sujeito a inundações sazonais, e as duas espécies foram submetidas às mesmas condições climáticas. Em Commelina diffusa foram encontradas larvas da família Agromyzidae e, em Floscopa glabrata observaram-se três exuvias cefálicas de Chironomidae. Os dados anat

  13. Experimental Evaluation of the Pathogenicity of Different Strains of Schistosoma mansoni

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    Antônio Aurélio Euzébio

    2012-01-01

    Full Text Available The pathogenesis of three different Schistosoma mansoni strains from the Brazilian states of Minas Gerais (BH strain and São Paulo (SJ and SD strains was evaluated in experimentally infected mice. Observations of the most severe clinical cases among local patients treated (SD strain in the city of Campinas (São Paulo, Brazil formed the basis of this study. Mice were used as definitive hosts and were infected with cercariae from Biomphalaria tenagophila (SJ and SD strains and Biomphalaria glabrata (BH strains. The parameters analyzed were as follows: number of S. mansoni eggs in mice feces; number of granulomas per tissue area in liver, spleen, lungs, pancreas, and ascending colon; measurements of hepatic and intestinal granulomas; number of adult worms; and measurements of trematode eggs. The comparison among the three strains indicated that the SD strain, isolated in Campinas, presented a higher worm recovery relative to the number of penetrating cercariae. In addition, when compared to the SJ and BH strains, the SD strain demonstrated similar pathogenicity to the BH strain, with a greater quantity of granulomas in the viscera, as well as larger granulomas and eggs. Furthermore, a greater quantity of trematode eggs was also shed in the feces.

  14. A low stringency polymerase chain reaction approach to the identification of Biomphalaria glabrata and B. tenagophila, intermediate snail hosts of Schistosoma mansoni in Brazil

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    Teofânia HDA Vidigal

    1996-12-01

    Full Text Available The low stringency-polymerase chain reaction (LS-PCR with a pair of specific primers for the amplification of the 18S rRNA gene was evaluated as a means of differentiating between the two Schistosoma mansoni intermediate host species in Brazil: Biomphalaria glabrata and B. tenagophila. Individual snails obtained from different states of Brazil were used and the amplification patterns obtained showed a high degree of genetic variability in these species. Nevertheless, 4 and 3 clearly defined specific diagnostic bands was observed in individuals from B. glabrata and B. tenagophila respectively. The detection of snail specific diagnostic bands suggests the possibility of reliable species differentiation at the DNA level using LS-PCR.

  15. Bioactivity Evaluation of Plant Extracts Used in Indigenous Medicine against the Snail, Biomphalaria glabrata, and the Larvae of Aedes aegypti.

    Science.gov (United States)

    Dos Santos, Edilson Alves; de Carvalho, Cenira M; Costa, Ana L S; Conceição, Adilva S; Moura, Flávia de B Prado; Santana, Antônio Euzébio Goulart

    2012-01-01

    This investigation examined the molluscicidal and larvicidal activity of eight plants that are used in the traditional medicine of the Pankararé indigenous people in the Raso da Catarina region, Bahia state, Brazil. The tested plants were chosen based on the results of previous studies. Only those plants that were used either as insect repellents or to treat intestinal parasitic infections were included in the study. Crude extracts (CEs) of these plants were tested for their larvicidal activity (against Aedes aegypti larvae in the fourth instar) and molluscicidal activity (against the snail Biomphalaria glabrata). The plant species Scoparia dulcis and Helicteres velutina exhibited the best larvicidal activities (LC(50) 83.426 mg/L and LC(50) 138.896 mg/L, resp.), and Poincianella pyramidalis, Chenopodium ambrosoides, and Mimosa tenuiflora presented the best molluscicidal activities (LC(50) 0.94 mg/L, LC(50) 13.51 mg/L, and LC(50) 20.22 mg/L, resp.). As we used crude extracts as the tested materials, further study is warranted to isolate and purify the most active compounds.

  16. Bioactivity Evaluation of Plant Extracts Used in Indigenous Medicine against the Snail, Biomphalaria glabrata, and the Larvae of Aedes aegypti

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    Edilson Alves dos Santos

    2012-01-01

    Full Text Available This investigation examined the molluscicidal and larvicidal activity of eight plants that are used in the traditional medicine of the Pankararé indigenous people in the Raso da Catarina region, Bahia state, Brazil. The tested plants were chosen based on the results of previous studies. Only those plants that were used either as insect repellents or to treat intestinal parasitic infections were included in the study. Crude extracts (CEs of these plants were tested for their larvicidal activity (against Aedes aegypti larvae in the fourth instar and molluscicidal activity (against the snail Biomphalaria glabrata. The plant species Scoparia dulcis and Helicteres velutina exhibited the best larvicidal activities (LC50 83.426 mg/L and LC50 138.896 mg/L, resp., and Poincianella pyramidalis, Chenopodium ambrosoides, and Mimosa tenuiflora presented the best molluscicidal activities (LC50 0.94 mg/L, LC50 13.51 mg/L, and LC50 20.22 mg/L, resp.. As we used crude extracts as the tested materials, further study is warranted to isolate and purify the most active compounds.

  17. Transcription factors Asg1p and Hal9p regulate pH homeostasis in Candida glabrata

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    Jing eWu

    2015-08-01

    Full Text Available Candida glabrata is an important microorganism used in commercial fermentation to produce pyruvate, but very little is known about its mechanisms for surviving acid stress in culture. In this study, it was shown that transcription factors Asg1p and Hal9p play essential roles in C. glabrata in the tolerance of acid stress, as the deletion of CgASG1 or CgHAL9 resulted in the inability to survive in an acidic environment. Cgasg1 and Cghal9 mutant strains are unable to maintain pH homeostasis, as evidenced by a decrease in intracellular pH and an increase in reactive oxygen species production, which results in metabolic disorders. The results showed that intracellular acidification was partly due to the diminished activity of the plasma membrane proton pump, CgPma1p. In addition, transcriptome sequencing revealed that Cgasg1 and Cghal9 mutant strains displayed a variety of changes in gene expression under acidic conditions, including genes in the MAPK signaling pathway, plasma membrane or cell wall organization, trehalose accumulation, and the RIM101 signaling pathway. Lastly, quantitative reverse-transcribed PCR and cellular localization showed that CgAsg1p and CgHal9p played independent roles in response to acid stress.

  18. Trypanosoma cruzi infection in the opossum Didelphis marsupialis: absence of neonatal transmission and protection by maternal antibodies in experimental infections

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    Ana M. Jansen

    1994-03-01

    Full Text Available The high rate of natural Trypanosoma cruzi infection found in opossums does not always correlate with appreciable densities of local triatomid populations. One alternative method which might bypass the invertebrate vector is direct transmission from mother to offspring. This possibility was investigated in five T. cruzi infected females and their litters (24 young. The influence of maternal antibodies transferred via lactation, on the course of experimental infection, was also examined. Our results show that neonatal transmission is probably not responsible for the high rate of natural T. cruzi infection among opossums. In addition antibodies of maternal origin confer a partial protection to the young. This was demonstrated by the finding of a double prepatency period and 4,5 fold lower levels of circulating parasites, in experimentally infected pouch young from infected as compared to control uninfected mothes. On the other hand, the duration of patent parasitemia was twice as long as that observed in the control group.

  19. Analysis of toxicity of Anacardium occidentale L. extract submitted to ionizing radiation on embryos of Biomphalaria glabrata and Artemia salina

    International Nuclear Information System (INIS)

    Silva, Hianna A.M.F.; Sa, Jose L.F.; Lima, Claudia S.A.; Amancio, Francisco F.; Melo, Ana M.M.A.; Ribeiro, Luanna R.S.; Santos, Gustavo H.F.; Silva, Edvane B.

    2013-01-01

    The use of gamma radiation as a sterilization method for herbs, herbal medicines and foods, shows positive results regarding the retention of such products, economy and safety of the method. However, it is known that this method of processing plant material can cause chemical changes in these products related to the type of material, its components and the dose received. Evaluated, in the present study, the action of gamma radiation as a modifier of toxicity extract of Anacardium occidentale Linn. To evaluate the toxicity of the extract irradiated at doses of 5.0, 7.5 and 10.0 kGy and concentrations of 250, 500 and 1000 mg/L was used bioassays with Artemia salina and Biomphalaria glabrata. For the test to A. salina, 520 specimens were used divided into groups of 10 larvae. For the bioassay with B. glabrata, 3900 specimens were used divided into groups of, approximately, 100 embryos. Larvae of A. salina and embryos were subjected to extracts irradiated and unirradiated for 24 hours. The bioassay with A. salina, showed a decrease, compared to extract unirradiated and irradiated at doses of 5.0 and 7.5 kGy, of extract irradiated with 10 kGy, where the mortality did not differ from the control group. In tests with embryos was observed an increase in the toxicity of the extract at a dose of 7.5 kGy and a decrease in the dose of 10.0 kGy. The radiation promoted changes in the toxicity of leaves extracts of Anacardium occidentale Linn. on embryos of Biomphalaria glabrata and Artemia salina. (author)

  20. Analysis of toxicity of Anacardium occidentale L. extract submitted to ionizing radiation on embryos of Biomphalaria glabrata and Artemia salina

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Hianna A.M.F.; Sa, Jose L.F.; Lima, Claudia S.A.; Amancio, Francisco F.; Melo, Ana M.M.A., E-mail: hiannaamfs@gmail.com, E-mail: luismuma6@gmail.com, E-mail: claudia.salima@gmail.com, E-mail: amancioff@bol.com.br, E-mail: amdemelo@hotmail.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Biofisica e Radiobiologia; Ribeiro, Luanna R.S.; Santos, Gustavo H.F.; Silva, Edvane B., E-mail: luannaribeiro_lua@hotmail.com, E-mail: santosghf@hotmail.com, E-mail: edvborges@yahoo.com.br [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Energia Nuclear. Grupo de Estudos em Radioprotecao e Radioecologia

    2013-07-01

    The use of gamma radiation as a sterilization method for herbs, herbal medicines and foods, shows positive results regarding the retention of such products, economy and safety of the method. However, it is known that this method of processing plant material can cause chemical changes in these products related to the type of material, its components and the dose received. Evaluated, in the present study, the action of gamma radiation as a modifier of toxicity extract of Anacardium occidentale Linn. To evaluate the toxicity of the extract irradiated at doses of 5.0, 7.5 and 10.0 kGy and concentrations of 250, 500 and 1000 mg/L was used bioassays with Artemia salina and Biomphalaria glabrata. For the test to A. salina, 520 specimens were used divided into groups of 10 larvae. For the bioassay with B. glabrata, 3900 specimens were used divided into groups of, approximately, 100 embryos. Larvae of A. salina and embryos were subjected to extracts irradiated and unirradiated for 24 hours. The bioassay with A. salina, showed a decrease, compared to extract unirradiated and irradiated at doses of 5.0 and 7.5 kGy, of extract irradiated with 10 kGy, where the mortality did not differ from the control group. In tests with embryos was observed an increase in the toxicity of the extract at a dose of 7.5 kGy and a decrease in the dose of 10.0 kGy. The radiation promoted changes in the toxicity of leaves extracts of Anacardium occidentale Linn. on embryos of Biomphalaria glabrata and Artemia salina. (author)

  1. Resistência de Biomphalaria peregrina de Santa Rita do Sapucaí, Minas Gerais, a infecção com três cepas de Schistosoma mansoni Resistance of Biomphalaria peregrina from Santa Rita do Sapucaí, State of Minas Gerais, Brazil, to infection with strain of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Cecília Pereira de Souza

    1988-12-01

    Full Text Available Descendentes do planorbídeo Biomphalaria peregrina, coletados em Santa Rita do Sapucaí, Minas Gerais, Brasil, foram expostos a miracídios de três cepas de Schistosoma mansoni: "LE" de Belo Horizonte, MG; "SJ" de São José dos Campos, SP e "AL" do Estado de Alagoas. Dentre 300 exemplares expostos, nenhum se infectou com as três cepas do trematódeo. Por outro lado, 300 exemplares de B. glabrata, dos grupos de controle, apresentaram taxas de infecção de 61,1 a 95,3% com as três cepas do trematódeo. As taxas de mortalidade de B. peregrina e de B. glabrata foram de 20,0 e de 28,0%, respectivamente.The descendants of the planorbid snail Biomphalaria peregrina, collected in the region of Santa rita do Sapucaí, Minas Gerais, Brazil, were exposed to miracidia of three strains of Schistosoma mansoni: "LE" strain from Belo Horizonte, State of Minas Gerais; "SJ", strain from São José dos Campos, State of São Paulo and "AL" strain from State of Alagoas. Of 300 snails exposed to miracidia of the three strains, none was infected. On the other hand, 300 Biomphalaria glabrata of the control groups showed infection rates of 61.1 to 95.3% with three strains. The mortality rates of B. peregrina and B. glabrata were 20% and 28%, respectively.

  2. Experimental infection of Artibeus intermedius with a vampire bat rabies virus.

    Science.gov (United States)

    Obregón-Morales, Cirani; Aguilar-Setién, Álvaro; Perea Martínez, Leonardo; Galvez-Romero, Guillermo; Martínez-Martínez, Flor Olivia; Aréchiga-Ceballos, Nidia

    2017-06-01

    Experimental infection of Artibeus intermedius, the great fruit-eating bat, was performed with vampire bat rabies isolates. Bats (n=35) were captured in the wild and quarantined prior to experimental infection. No rabies antibodies were detected by rapid fluorescent focus inhibition test (RFFIT) prior to infection. Three doses of rabies virus (RV) and three different routes of infection were used. One out of 35 bats died without showing any clinical signs at day 14 and was positive for rabies. None of the 34 other bats showed clinical signs for rabies, but high antibody titers were detected post-inoculation, suggesting either innate immune response to the vampire bat rabies virus or possible pre-exposure to RV and inoculation leading to a booster effect. Rabies virus was detected by hemi-nested RT-PCR (hnRT-PCR) in the brain (n=3), stomach (n=1) of bats that were negative by immunofluorescence and that survived rabies infection. The bat that died on day 14 was positive by hnRT-PCR on the brain, heart and liver. These results suggest that either previous non-lethal exposure to RV or natural low susceptibility to vampire bat viruses somehow protected Artibeus intermedius from clinical rabies infection leading to a marginal lethality effect on this bats species population in the wild. Copyright © 2017 Elsevier Ltd. All rights reserved.

  3. Genome-Wide Scan and Test of Candidate Genes in the Snail Biomphalaria glabrata Reveal New Locus Influencing Resistance to Schistosoma mansoni.

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    Jacob A Tennessen

    Full Text Available New strategies to combat the global scourge of schistosomiasis may be revealed by increased understanding of the mechanisms by which the obligate snail host can resist the schistosome parasite. However, few molecular markers linked to resistance have been identified and characterized in snails.Here we test six independent genetic loci for their influence on resistance to Schistosoma mansoni strain PR1 in the 13-16-R1 strain of the snail Biomphalaria glabrata. We first identify a genomic region, RADres, showing the highest differentiation between susceptible and resistant inbred lines among 1611 informative restriction-site associated DNA (RAD markers, and show that it significantly influences resistance in an independent set of 439 outbred snails. The additive effect of each RADres resistance allele is 2-fold, similar to that of the previously identified resistance gene sod1. The data fit a model in which both loci contribute independently and additively to resistance, such that the odds of infection in homozygotes for the resistance alleles at both loci (13% infected is 16-fold lower than the odds of infection in snails without any resistance alleles (70% infected. Genome-wide linkage disequilibrium is high, with both sod1 and RADres residing on haplotype blocks >2 Mb, and with other markers in each block also showing significant effects on resistance; thus the causal genes within these blocks remain to be demonstrated. Other candidate loci had no effect on resistance, including the Guadeloupe Resistance Complex and three genes (aif, infPhox, and prx1 with immunological roles and expression patterns tied to resistance, which must therefore be trans-regulated.The loci RADres and sod1 both have strong effects on resistance to S. mansoni. Future approaches to control schistosomiasis may benefit from further efforts to characterize and harness this natural genetic variation.

  4. Reduction in transmission of Schistosoma mansoni by a four-year focal mollusciciding programme against Biomphalaria glabrata in Saint Lucia.

    Science.gov (United States)

    Prentice, M A; Jordan, P; Bartholomew, R K; Grist, E

    1981-01-01

    The effect of transmission of Schistosoma mansoni of a focal snail control programme was investigated over four years amongst approximately 1250 people living in five communities in the steep-sided Soufriere river valley, St. Lucia, West Indies. Bayer 6076 was applied from constant flow drip cans to 12 stream sections at a target dose of 8 mg/litre clonitralide every four weeks. Only proven and potential transmission sites were treated; marsh habitats, where Biomphalaria glabrata were widespread, were ignored. In the stream snail numbers were reduced by 94% in the first year and by 100% thereafter. Incidence of new S. mansoni infections amongst children fell from 18% in the last year before control to 6% and 9% after three and four years respectively. Amongst children and adults in the four years of control the conversion/reversion ratio declined leading to a lowering of the over-all prevalence from 40% to 22%. Parasitologically the results were similar to those of a previously evaluated area-wide mollusciciding programme. The mean annual cost per person protected was US $2.60. This figure is atypically high because the topography of the area severely limited the population size.

  5. Evaluation of potassium permanganate against an experimental subacute infection of Flavobacterium columnare in channel catfish, Icatlurus punctatus

    Science.gov (United States)

    The efficacy of potassium permanganate (KMnO4) as a prophylactic and therapeutic treatment for subacute infection of Flavobacterium columnare was demonstrated in experimentally infected channel catfish, Ictalurus punctatus. Catfish experimentally infected with F. columnare to mimic a subacute infec...

  6. Experimental Salmonella typhimurium infections in rats. I

    DEFF Research Database (Denmark)

    Hougen, H P; Jensen, E T; Klausen, B

    1989-01-01

    The course of experimentally induced Salmonella typhimurium infection was studied in three groups of inbred LEW rats: homozygous +/+, athymic rnu/rnu and isogeneic thymus-grafted rnu/rnu rats. In the first experiment the animals were inoculated intraperitoneally with 10(8) bacteria and all animals...... became severely septicemic and died within a week of inoculation, irrespective of presence or absence of thymus. In the second experiment the animals were inoculated with 10(6) bacteria, and both euthymic and thymus-grafted animals responded with high titres of anti bacterial antibodies while these were...... very low in the athymic nude animals. Polyclonal antibody production was only observed in the euthymic animals and only regarding IgG. Athymic rats were not able to clear the infection, while the thymus-grafted animals reacted like euthymic rats: Very few animals housed the bacteria four weeks after...

  7. A Targeted Capture Linkage Map Anchors the Genome of the Schistosomiasis Vector Snail, Biomphalaria glabrata.

    Science.gov (United States)

    Tennessen, Jacob A; Bollmann, Stephanie R; Blouin, Michael S

    2017-07-05

    The aquatic planorbid snail Biomphalaria glabrata is one of the most intensively-studied mollusks due to its role in the transmission of schistosomiasis. Its 916 Mb genome has recently been sequenced and annotated, but it remains poorly assembled. Here, we used targeted capture markers to map over 10,000 B. glabrata scaffolds in a linkage cross of 94 F1 offspring, generating 24 linkage groups (LGs). We added additional scaffolds to these LGs based on linkage disequilibrium (LD) analysis of targeted capture and whole-genome sequences of 96 unrelated snails. Our final linkage map consists of 18,613 scaffolds comprising 515 Mb, representing 56% of the genome and 75% of genic and nonrepetitive regions. There are 18 large (> 10 Mb) LGs, likely representing the expected 18 haploid chromosomes, and > 50% of the genome has been assigned to LGs of at least 17 Mb. Comparisons with other gastropod genomes reveal patterns of synteny and chromosomal rearrangements. Linkage relationships of key immune-relevant genes may help clarify snail-schistosome interactions. By focusing on linkage among genic and nonrepetitive regions, we have generated a useful resource for associating snail phenotypes with causal genes, even in the absence of a complete genome assembly. A similar approach could potentially improve numerous poorly-assembled genomes in other taxa. This map will facilitate future work on this host of a serious human parasite. Copyright © 2017 Tennessen et al.

  8. Effect of Exposure Dose on Ichthyophonus Prevalence and Infection Intensity in Experimentally Infected Rainbow Trout, Oncorhynchus mykiss.

    Science.gov (United States)

    Kocan, Richard; LaPatra, Scott

    2016-02-01

    This study describes the effect of increasing exposure dose on Ichthyophonus prevalence and infection intensity in experimentally infected rainbow trout, Oncorhynchus mykiss. Specific-pathogen free trout were exposed per os to increasing numbers of Ichthyophonus schizonts obtained from naturally infected donor fish, then sampled after 30 and 60 days post-exposure. Both in vitro explant culture and histology revealed that as the number of schizonts per dose increased there was a proportionate increase in the number of infected fish, as well as an increase in the number of infected organs; parasite density in individual infected organs also increased with dose. Explant culture revealed that all fish exposed to the highest dose (≥2,080 schizonts) became infected, while only 67% of those exposed to the intermediate dose (1,040-1,153 schizonts) were Ichthyophonus-positive after 60 days; Ichthyophonus was not detected in fish exposed to the 2 lowest doses (≤280 schizonts). Histologic examination of individual infected organs also revealed increasing infection prevalence and parasite density in response to exposure to increasing numbers of Ichthyophonus schizonts.

  9. Silver colloidal nanoparticles: effect on matrix composition and structure of Candida albicans and Candida glabrata biofilms.

    Science.gov (United States)

    Monteiro, D R; Silva, S; Negri, M; Gorup, L F; de Camargo, E R; Oliveira, R; Barbosa, D B; Henriques, M

    2013-04-01

    The aim of this study was to assess the effect of different silver nanoparticles (SN) concentrations on the matrix composition and structure of Candida albicans and Candida glabrata biofilms. Candida biofilms were developed in 6-well microtiter plates during 48 h. After, these biofilms were exposed to 13.5 or 54 μg SN ml(-1) for 24 h. Then, extracellular matrices were extracted from biofilms and analysed chemically in terms of proteins, carbohydrates and DNA. To investigate the biofilm structure, scanning electron microscopy (SEM) and epifluorescence microscopy were used. SN interfered with the matrix composition of Candida biofilms tested in terms of protein, carbohydrate and DNA, except for the protein content of C. albicans biofilm. By SEM, Candida biofilms treated with SN revealed structural differences, when compared with the control groups. Further, SN showed a trend of agglomeration within the biofilms. Epifluorescence microscopy images suggest that SN induced damage on cell walls of the Candida isolates tested. In general, irrespective of concentration, SN affected the matrix composition and structure of Candida biofilms and these findings may be related to the mechanisms of biocide action of SN. This study reveals new insights about the behaviour of SN when in contact with Candida biofilms. SN may contribute to the development of therapies to prevent or control Candida infections. © 2012 The Society for Applied Microbiology.

  10. Prevalence, virulence factors and antifungal susceptibility of Candida spp. isolated from bloodstream infections in a tertiary care hospital in Brazil.

    Science.gov (United States)

    Canela, Heliara Maria Spina; Cardoso, Bárbara; Vitali, Lucia Helena; Coelho, Harnoldo Colares; Martinez, Roberto; Ferreira, Márcia Eliana da Silva

    2018-01-01

    Candida spp. are responsible for 80% of all systemic fungal infections and are associated with high mortality rates. This study characterised 79 bloodstream isolates of C. albicans, C. glabrata, C. orthopsilosis, C. parapsilosis and C. tropicalis from patients in a Brazilian hospital. The susceptibility to amphotericin B, caspofungin, fluconazole and voriconazole was determined; virulence factor production was assessed based on haemolysin, phospholipase and proteinase activities, and the patients' clinical characteristics were analysed. C. albicans was the predominant species (44%), followed by C. glabrata (19%), C. tropicalis (19%), C. parapsilosis (14%) and C. orthopsilosis (4%). The candidemia incidence was 1.52 per 1000 admissions, and the crude mortality rate was 52%. One C. albicans isolate was resistant to fluconazole and voriconazole. Moreover, 20.2%, 2.5% and 3.8% of the isolates exhibited dose-dependent susceptibility to fluconazole, voriconazole and caspofungin, respectively. In conclusion, although the C. glabrata incidence was higher than that usually described in Brazil, its increase was previously observed in studies conducted worldwide. Furthermore, the azole resistance of the C. albicans isolate could be due to previous exposure to these antifungals. These results highlight the importance of epidemiological studies and will facilitate an improved understanding of candidemia in the studied hospital. © 2017 Blackwell Verlag GmbH.

  11. Evaluation of a simple Theileria annulata culture protocol from experimentally infected bovine whole blood

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    Gharbi M.

    2012-08-01

    Full Text Available We have evaluated a new simple technique using whole blood from experimentally infected cattle for the isolation and cultivation of Theileria annulata. The study was carried out on 20 Holstein-Frisian bovines that had been experimentally infected with a virulent lethal dose of Theileria annulata. This technique has been compared to the classical peripheral blood monocyte isolation with Ficoll carried out on 22 experimentally infected Holstein-Friesian calves. The effectiveness of the reference technique was estimated to 86.4%, whilst the effectiveness of the new technique was 100%. Moreover, this new technique leads to time and money saving estimated to € 3.06 per sample. It decreases the contamination risks by reducing the steps of sample manipulation.

  12. Suppression of inflammatory immune responses in celiac disease by experimental hookworm infection.

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    Henry J McSorley

    Full Text Available We present immunological data from two clinical trials where the effect of experimental human hookworm (Necator americanus infection on the pathology of celiac disease was evaluated. We found that basal production of Interferon- (IFN-γ and Interleukin- (IL-17A from duodenal biopsy culture was suppressed in hookworm-infected participants compared to uninfected controls. Increased levels of CD4+CD25+Foxp3+ cells in the circulation and mucosa are associated with active celiac disease. We show that this accumulation also occurs during a short-term (1 week oral gluten challenge, and that hookworm infection suppressed the increase of circulating CD4+CD25+Foxp3+ cells during this challenge period. When duodenal biopsies from hookworm-infected participants were restimulated with the immunodominant gliadin peptide QE65, robust production of IL-2, IFN-γ and IL-17A was detected, even prior to gluten challenge while participants were strictly adhering to a gluten-free diet. Intriguingly, IL-5 was produced only after hookworm infection in response to QE65. Thus we hypothesise that hookworm-induced TH2 and IL-10 cross-regulation of the TH1/TH17 inflammatory response may be responsible for the suppression of these responses during experimental hookworm infection.

  13. Serum levels of cytokines in water buffaloes experimentally infected with Fasciola gigantica.

    Science.gov (United States)

    Zhang, Fu-Kai; Guo, Ai-Jiang; Hou, Jun-Ling; Sun, Miao-Miao; Sheng, Zhao-An; Zhang, Xiao-Xuan; Huang, Wei-Yi; Elsheikha, Hany M; Zhu, Xing-Quan

    2017-09-15

    Fasciola gigantica infection in water buffaloes causes significant economic losses especially in developing countries. Although modulation of the host immune response by cytokine neutralization or vaccination is a promising approach to control infection with this parasite, our understanding of cytokine's dynamic during F. gigantica infection is limited. To address this, we quantified the levels of serum cytokines produced in water buffaloes following experimental infection with F. gigantica. Five buffaloes were infected via oral gavage with 500 viable F. gigantica metacercariae and blood samples were collected from buffaloes one week before infection and for 13 consecutive weeks thereafter. The levels of 10 cytokines in serum samples were simultaneously determined using ELISA. F. gigantica failed to elicit the production of various pro-inflammatory cytokines, including interleukin-1β (IL-1β), IL-2, IL-6, IL-12, and IFN-γ. On the other hand, evidence of a Th2 type response was detected, but only early in the course of parasite colonization and included modest increase in the levels of IL-10 and IL-13. The results also revealed suppression of the immune responses as a feature of chronic F. gigantica infection in buffaloes. Taken together, F. gigantica seems to elicit a modest Th2 response at early stage of infection in order to downregulate harmful Th1- and Th17-type inflammatory responses in experimentally infected buffaloes. The full extent of anti-F. gigantica immune response and its relation to pathogenesis requires further study. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Candida bloodstream infection: a clinical microbiology laboratory perspective.

    Science.gov (United States)

    Pongrácz, Júlia; Kristóf, Katalin

    2014-09-01

    The incidence of Candida bloodstream infection (BSI) has been on the rise in several countries worldwide. Species distribution is changing; an increase in the percentage of non-albicans species, mainly fluconazole non-susceptible C. glabrata was reported. Existing microbiology diagnostic methods lack sensitivity, and new methods need to be developed or further evaluation for routine application is necessary. Although reliable, standardized methods for antifungal susceptibility testing are available, the determination of clinical breakpoints remains challenging. Correct species identification is important and provides information on the intrinsic susceptibility profile of the isolate. Currently, acquired resistance in clinical Candida isolates is rare, but reports indicate that it could be an issue in the future. The role of the clinical microbiology laboratory is to isolate and correctly identify the infective agent and provide relevant and reliable susceptibility data as soon as possible to guide antifungal therapy.

  15. Imaging experimental infective endocarditis with indium-111-labeled blood cellular components

    International Nuclear Information System (INIS)

    Riba, A.L.; Thakur, M.L.; Gottschalk, A.; Andriole, V.T.; Zaret, B.L.

    1979-01-01

    The capability of radionuclide imaging to detect experimental aortic valve infective endocarditis was assessed with indium-111 ( 111 In)-labeled blood cells. Sequential cardiac imaging and tissue distribution studies were obtained in 17 rabbits with infective endocarditis after administration of 111 In-platelets and in five after 111 In-polymorphonuclear leukocytes. Forty-eight to 72 hours after platelet administration, in vivo imaging demonstrated abnormal 111 In uptake in all animals in the region of the aortic valve in an anatomically distinct pattern. Images of the excised heart showed discrete cardiac uptake conforming to the in vivo image and gross pathological examination. 111 In-platelet uptake in vegetations from the 17 animals averaged 240 +- 41 times greater than that in normal myocardium and 99 +- 15 times greater uptake in blood. In contrast, 111 In-leukocyte cardiac imaging showed no abnormal aortic valve uptake 24 hours after tracer administration and the lesion myocardium activity ratio was only 5 +- 2 (3 +- 1 for lesion/blood activity). Four normal rabbits demonstrated neither positive 111 In-platelet scintigraphs nor abnormal cardiac tissue uptake. Likewise, noncellular 111 In was not concentrated to any significant extent in three animals with infective endocarditis. This study demonstrates that 111 In-platelet, but not leukocyte cardiac imaging, is a sensitive technique for detecting experimental infective endocarditis. The imaging data conform to the cellular pathology of the infective endocarditis vegetation

  16. The dual role of Candida glabrata Drug:H+ Antiporter CgAqr1 (ORF CAGL0J09944g in antifungal drug and acetic acid resistance

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    Catarina eCosta

    2013-06-01

    Full Text Available Opportunistic Candida species often have to cope with inhibitory concentrations of acetic acid, in the acidic environment of the vaginal mucosa. Given that the ability of these yeast species to tolerate stress induced by weak acids and antifungal drugs appears to be a key factor in their persistence and virulence, it is crucial to understand the underlying mechanisms.In this study, the Drug:H+ Antiporter CgAqr1 (ORF CAGL0J09944g, from Candida glabrata, was identified as a determinant of resistance to acetic acid, and also to the antifungal agents flucytosine and, less significantly, clotrimazole. These antifungals were found to act synergistically with acetic acid against this pathogen. The action of CgAqr1 in this phenomenon was analyzed. Using a GFP fusion, CgAqr1 was found to localize to the plasma membrane and to membrane vesicles when expressed in C. glabrata or, heterologously, in Saccharomyces cerevisiae. Given its ability to complement the susceptibility phenotype of its S. cerevisiae homolog, ScAqr1, CgAqr1 was proposed to play a similar role in mediating the extrusion of chemical compounds. Significantly, the expression of this gene was found to reduce the intracellular accumulation of 3H-flucytosine and, to a moderate extent, of 3H-clotrimazole, consistent with a direct role in antifungal drug efflux. Interestingly, no effect of CgAQR1 deletion could be found on the intracellular accumulation of 14C-acetic acid, suggesting that its role in acetic acid resistance may be indirect, presumably through the transport of a still unidentified physiological substrate. Although neither of the tested chemicals induces changes in CgAQR1 expression, pre-exposure to flucytosine or clotrimazole was found to make C. glabrata cells more sensitive to acetic acid stress. Results from this study show that CgAqr1 is an antifungal drug resistance determinant and raise the hypothesis that it may play a role in C. glabrata persistent colonization and

  17. Susceptibility of Biomphalaria tenagophila and Biomphalaria straminea to Schistosoma mansoni infection detected by low stringency polymerase chain reaction

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    JANNOTTI-PASSOS Liana Konovaloff

    2000-01-01

    Full Text Available In order to determine Schistosoma mansoni infection rates in Biomphalaria tenagophila and B. straminea, low stringency polymerase chain reaction (LS-PCR technique was used as a complementary method to light exposure technique. LS-PCR has already been standardized in our laboratory to detect the trematode DNA in B. glabrata. Higher S. mansoni infection rates were detected using conventional method and LS-PCR. The parasite DNA profile was detected in both species after 7-day exposure to miracidia, using LS-PCR. This technique enables early detection of schistosomiasis transmission focuses, in endemic areas, before the beginning of cercariae shedding.

  18. Effects of 60Co gamma radiation on Biomphalaria glabrata (Say, 1818) Embryo, I. Mortality

    International Nuclear Information System (INIS)

    Okazaki, K.; Kawano, T.

    1990-01-01

    A study was conducted on the radiosensitivity of Biomphalaria glabrata embryos submitted to doses of 5, 10,15,20 and 25 Gy of 60 Co during the cleavage, blastula, gastrula, young trochophore and trochophore stages. Mortality was the parameter used to evaluated the damage induced by ionizing radiation. Susceptibility decreased with increasing embryo age and with decreased radiation dose. Estimated LD 50 values (15 days) showed that the cleavage stage (4.3 Gy) was approximately four times more radiosensitive than the trochophore stage (17.0 Gy). The survival curves obtained for each embryo stage are discussed on the basis of the multitarget theory. (author) [pt

  19. The pathogenesis of single experimental infections with Strongylus vulgaris in foals.

    Science.gov (United States)

    Duncan, J L; Pirie, H M

    1975-01-01

    The clinical signs, pathology and clinical pathology associated with single experimental infections of Strongylus vulgaris in worm-free pony foals are described. The major clinical signs which became apparent in the infected foals during the first three weeks were pyrexia, anorexia, dullness and abdominal pain. Within the first two weeks of infection lesions were confined to the intestine and terminal branches of the intestinal arteries and consisted of mucosal, submucosal and serosal haemorrhage together with arteritis of submucosal and serosal arteries and also a marked inflammatory reaction. The main lesion seen three weeks after infection was gross thrombosis of the anterior mesenteric artery or one of its major branches. On section these affected arteries showed marked intimal thickening with infiltration of plasma cells, lymphocytes, macrophages and neutrophils. Between one and four months after infection the gross lesions were predominantly in the arteries and consisted of fibrous thickening of the arterial wall and thrombosis associated with the presence of developing fourth stage larvae. Four months after infection the arterial lesions were still prominent and microscopically there was fibrosis of the wall of the affected artery with wide-spread disruption of the intima. In the adventitia organised thrombi were apparent in the vasa vasorum and resulted in the obliteration of their lumina. The typical lesion associated with the return of fifth stage larvae to the intestine was nodule formation in close proximity to thrombosed terminal intestinal arteries and sections of parasites were seen in the intestinal wall surrounded by neutrophils and necrotic debris. By nine months after infection the arterial lesion had healed, but histologically there was fibrosis of the intima and macrophages containing haemosiderin were seen in the arterial wall. The most significant haematological findings during the experimental period were a marked polymorphonuclear

  20. The introduction of Melanoides tuberculata (Mollusca: Thiaridae) to the island of Saint Lucia (West Indies) and its role in the decline of Biomphalaria glabrata, the snail intermediate host of Schistosoma mansoni.

    Science.gov (United States)

    Pointier, J P

    1993-06-01

    A malacological survey was carried out in May 1992 in the whole hydrographic system of Saint Lucia 11 years after the end of a biological control programme to eliminate Biomphalaria glabrata, the snail intermediate host of Schistosoma mansoni. A competitor snail, Melanoides tuberculata, was introduced to Saint Lucia in 1978 and field experiments in several habitats were conducted by Prentice between 1978 and 1986. At the present time M. tuberculata is the most common freshwater snail in Saint Lucia. The results of the survey, undertaken in sites where B. glabrata occurred in large populations in the past showed (i) the absence of the snail hosts from seven sites now extensively colonized by the competitor (ii) the presence of B. glabrata in low or very low densities in 17 sites together with the competitor and (iii) the presence of the intermediate hosts in large populations in only two sites where M. tuberculata was absent. These results confirm the positive results observed by Prentice. The presence of another planorbid snail, B. straminea, is reported for the first time in Saint Lucia.

  1. Aspectos ultraestruturais de hemócitos de Biomphalaria glabrata Say (1818 (Gastropoda: Planorbidae analisados sob microscopia eletrônica de transmissão Ultrastructural aspects of hemocytes from Biomphalaria glabrata Say (1818 (Gastropoda: Planorbidae analysed with transmission eletronic microscopy

    Directory of Open Access Journals (Sweden)

    Marco Antonio Vasconcelos Santos

    2009-09-01

    Full Text Available Os hemócitos do caramujo Biomphalaria glabrata, um importante transmissor do trematódeo Schistosoma mansoni no Brasil, foram coletados de especimens na região Bragantina, localizada a oeste do estado do Pará. Os hemócitos foram examinados por meio de microscopia eletrônica de transmissão. As células foram fixadas pelo método de rotina com o uso do tampão PHEM (PIPES, HEPES, EGTA e Magnésio. Foram descritos os aspectos ultra-estruturais celulares como inclusões citoplasmáticas limitadas por membranas, mitocôndrias, retículos endoplasmáticos e outros. As observações mostram que esse tampão possui a propriedade de preservação do citoesqueleto celular, apresentando bons resultados na preservação das estruturas dos hemócitos e suas organelas.The blood cells of the pulmonate snail Biomphalaria glabrata from the region of Bragantina in the state of Pará, an important vector of the trematode Schistosoma mansoni in Brazil, were collected and fixed by routine method with PHEM buffer and examined with transmission electron microscopy. Ultrastructural cellular aspects like cytoplasmic inclusions, mitochondrion, vesicles and others organelles are described. The buffer used resulted in good preservations of the hemocytes and their organelles.

  2. Efficacy of imidocarb dipropionate in eliminating Theileria equi from experimentally infected horses.

    Science.gov (United States)

    Grause, Juanita F; Ueti, Massaro W; Nelson, Jeffrey T; Knowles, Donald P; Kappmeyer, Lowell S; Bunn, Thomas O

    2013-06-01

    Theileria equi, one of the causative agents of equine piroplasmosis, is endemic in many regions of the world but is considered a 'foreign' animal disease in the USA. In an effort to prevent the importation of T. equi, stringent serological screening of horses is practiced prior to entry to the USA. Current regulatory options available where horses are found to be infected include permanent quarantine with or without chemotherapy, repatriation, or euthanasia. Chemotherapeutics that eliminate infection and subsequently transmission risk are critical in the management of infected horses. In this study, the efficacy of the drug imidocarb dipropionate against experimental T. equi infection was assessed. Of nine horses experimentally inoculated with T. equi isolated from an animal previously imported from Peru, six were treated with imidocarb dipropionate after the resolution of the acute phase of the disease. Elimination of the parasite was demonstrated in 5/6 by nested PCR, blood transfusions to naïve horses, and reversion to seronegative status. The findings support the use of this drug as a potential treatment option in controlling outbreaks of T. equi, and also suggest that 'combination testing' using both serological and PCR detection methods are necessary to demonstrate clearance of infection. Published by Elsevier Ltd.

  3. KRE5 Suppression Induces Cell Wall Stress and Alternative ER Stress Response Required for Maintaining Cell Wall Integrity in Candida glabrata

    Science.gov (United States)

    Sasaki, Masato; Ito, Fumie; Aoyama, Toshio; Sato-Okamoto, Michiyo; Takahashi-Nakaguchi, Azusa; Chibana, Hiroji; Shibata, Nobuyuki

    2016-01-01

    The maintenance of cell wall integrity in fungi is required for normal cell growth, division, hyphae formation, and antifungal tolerance. We observed that endoplasmic reticulum stress regulated cell wall integrity in Candida glabrata, which possesses uniquely evolved mechanisms for unfolded protein response mechanisms. Tetracycline-mediated suppression of KRE5, which encodes a predicted UDP-glucose:glycoprotein glucosyltransferase localized in the endoplasmic reticulum, significantly increased cell wall chitin content and decreased cell wall β-1,6-glucan content. KRE5 repression induced endoplasmic reticulum stress-related gene expression and MAP kinase pathway activation, including Slt2p and Hog1p phosphorylation, through the cell wall integrity signaling pathway. Moreover, the calcineurin pathway negatively regulated cell wall integrity, but not the reduction of β-1,6-glucan content. These results indicate that KRE5 is required for maintaining both endoplasmic reticulum homeostasis and cell wall integrity, and that the calcineurin pathway acts as a regulator of chitin-glucan balance in the cell wall and as an alternative mediator of endoplasmic reticulum stress in C. glabrata. PMID:27548283

  4. Avaliação da bioatividade dos extratos de cúrcuma (Curcuma longa L., Zingiberaceae em Artemia salina e Biomphalaria glabrata

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    Carlos R. M. da Silva Filho

    Full Text Available A cúrcuma é o rizoma limpo, em boas condições, seco e moído da Curcuma longa L., uma planta herbácea da família Zingiberaceae. Visando novas alternativas para o controle da esquistossomose, os extratos de Curcuma longa L. foram testados para a avaliação da atividade moluscicida contra caramujos adultos da espécie Biomphalaria glabrata, e toxicidade (ensaio de letalidade com Artemia salina. A oleoresina e o óleo essencial de cúrcuma foram ativos contra Artemia salina (CL50 = 80,43 e CL50 = 319,82 μg/mL, respectivamente e também ativos contra os indivíduos adultos de Biomphalaria glabrata (CL50 = 58,3 e CL50 = 46,73 μg/mL, respectivamente. A partir dos resultados obtidos pôde ser concluído que ambos os extratos podem constituir uma alternativa no controle da população desses caramujos e na redução da esquistossomose.

  5. The Candida Pathogenic Species Complex

    Science.gov (United States)

    Turner, Siobhán A.; Butler, Geraldine

    2014-01-01

    Candida species are the most common causes of fungal infection. Approximately 90% of infections are caused by five species: Candida albicans, Candida glabrata, Candida tropicalis, Candida parapsilosis, and Candida krusei. Three (C. albicans, C. tropicalis, and C. parapsilosis) belong to the CTG clade, in which the CTG codon is translated as serine and not leucine. C. albicans remains the most commonly isolated but is decreasing relative to the other species. The increasing incidence of C. glabrata is related to its reduced susceptibility to azole drugs. Genome analysis suggests that virulence in the CTG clade is associated with expansion of gene families, particularly of cell wall genes. Similar independent processes took place in the C. glabrata species group. Gene loss and expansion in an ancestor of C. glabrata may have resulted in preadaptations that enabled pathogenicity. PMID:25183855

  6. Therapeutic effect of cefozopran (SCE-2787), a new parenteral cephalosporin, against experimental infections in mice.

    Science.gov (United States)

    Iizawa, Y; Okonogi, K; Hayashi, R; Iwahi, T; Yamazaki, T; Imada, A

    1993-01-01

    The therapeutic effect of cefozopran (SCE-2787), a new semisynthetic parenteral cephalosporin, against experimental infections in mice was examined. Cefozopran was more effective than cefpiramide and was as effective as ceftazidime and cefpirome against acute respiratory tract infections caused by Klebsiella pneumoniae DT-S. In the model of chronic respiratory tract infection caused by K. pneumoniae 27, cefozopran was as effective as ceftazidime. The therapeutic effect of cefozopran against urinary tract infections caused by Pseudomonas aeruginosa P9 was superior to that of cefpirome and was equal to those of ceftazidime and cefclidin. In addition, cefozopran was more effective than ceftazidime and was as effective as flomoxef in a thigh muscle infection caused by methicillin-sensitive Staphylococcus aureus 308A-1. Against thigh muscle infections caused by methicillin-resistant S. aureus N133, cefozopran was the most effective agent. The potent therapeutic effect of cefozopran in those experimental infections in mice suggests that it would be effective against respiratory tract, urinary tract, and soft tissue infections caused by a variety of gram-positive and gram-negative bacteria in humans. PMID:8431004

  7. Implications of water hardness in ecotoxicological assessments for water quality regulatory purposes: a case study with the aquatic snail Biomphalaria glabrata (Say, 1818

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    EC Oliveira-Filho

    Full Text Available Water hardness is a property depending on the presence of alkaline earth metals, mainly calcium and magnesium. Among the strategies for water quality monitoring, ecotoxicological assays are performed to minimize impacts and classify water bodies. For these laboratory evaluations parameters are previously defined in the guidelines, including water hardness for both cultivation and testing medium. The present work was performed to evaluate the effects of different levels of water hardness on the survival and reproduction of the freshwater snail Biomphalaria glabrata and discuss the influence of natural water hardness on the results of ecotoxicological tests with these environmental samples. Comparing the groups it was possible to observe that those maintained in waters with least hardness had lower reproductive success, while the groups maintained in highest hardness showed better reproduction. These data show that waters with low hardness make the reproduction of the snail B. glabrata unfeasible, and this reveal a problem for ecotoxicity assays using natural water samples.

  8. Immunologic responses in corn snakes (Pantherophis guttatus) after experimentally induced infection with ferlaviruses.

    Science.gov (United States)

    Neul, Annkatrin; Schrödl, Wieland; Marschang, Rachel E; Bjick, Tina; Truyen, Uwe; von Buttlar, Heiner; Pees, Michael

    2017-04-01

    OBJECTIVE To measure immunologic responses of snakes after experimentally induced infection with ferlaviruses. ANIMALS 42 adult corn snakes (Pantherophis guttatus) of both sexes. PROCEDURES Snakes were inoculated intratracheally with genogroup A (n = 12), B (12), or C (12) ferlavirus (infected groups) or cell-culture supernatant (6; control group) on day 0. Three snakes from each infected group were euthanized on days 4, 16, 28, and 49, and 3 snakes from the control group were euthanized on day 49. Blood samples were collected from live snakes on days -6 (baseline), 4, 16, 28, and 49. Hematologic tests were performed and humoral responses assessed via hemagglutination-inhibition assays and ELISAs. Following euthanasia, gross pathological and histologic evaluations and virus detection were performed. RESULTS Severity of clinical signs of and immunologic responses to ferlavirus infection differed among snake groups. Hematologic values, particularly WBC and monocyte counts, increased between days 4 and 16 after infection. A humoral response was identified between days 16 and 28. Serum IgM concentrations increased from baseline earlier than IgY concentrations, but the IgY relative increase was higher at the end of the study. The hemagglutination-inhibition assay revealed that the strongest reactions in all infected groups were against the strain with which they had been infected. Snakes infected with genogroup A ferlavirus had the strongest immune response, whereas those infected with genogroup B had the weakest responses. CONCLUSIONS AND CLINICAL RELEVANCE Results of this experimental study suggested that the ferlavirus strain with the highest virulence induced the weakest immune response in snakes.

  9. Characterising the mucosal and systemic immune responses to experimental human hookworm infection.

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    Soraya Gaze

    2012-02-01

    Full Text Available The mucosal cytokine response of healthy humans to parasitic helminths has never been reported. We investigated the systemic and mucosal cytokine responses to hookworm infection in experimentally infected, previously hookworm naive individuals from non-endemic areas. We collected both peripheral blood and duodenal biopsies to assess the systemic immune response, as well as the response at the site of adult worm establishment. Our results show that experimental hookworm infection leads to a strong systemic and mucosal Th2 (IL-4, IL-5, IL-9 and IL-13 and regulatory (IL-10 and TGF-β response, with some evidence of a Th1 (IFN-γ and IL-2 response. Despite upregulation after patency of both IL-15 and ALDH1A2, a known Th17-inducing combination in inflammatory diseases, we saw no evidence of a Th17 (IL-17 response. Moreover, we observed strong suppression of mucosal IL-23 and upregulation of IL-22 during established hookworm infection, suggesting a potential mechanism by which Th17 responses are suppressed, and highlighting the potential that hookworms and their secreted proteins offer as therapeutics for human inflammatory diseases.

  10. Detection of Toxoplasma gondii DNA in sera samples of mice experimentally infected

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    H. Langoni

    2006-04-01

    Full Text Available Detection of Toxoplasma gondii (T. gondii DNA in blood can help to diagnose the disease in its acute phase; however, it must be considered that hemoglobin, present in blood, can inhibit polymerase activity, making impracticable the detection of DNA in samples. Mice were experimentally infected via oral route with ME49 and BTU2 strains cysts and RH strain tachyzoites; polymerase chain reaction was used to detect T. gondii DNA in mice sera 18, 24, 48, 96, and 192 hours post infection (PI. Toxoplama gondii DNA was detected in only one animal infected with BTU2 strain, genotype III (isolated from a dog with neurological signs 18 hours PI. The agent's DNA was not detected in any sample of the other experimental groups. New studies must be carried out to verify the technique sensitivity in researches on this agent's genetic material using sera samples of acute-phase toxoplasmosis patients, especially in cases of immunosuppression.

  11. Estudo da potencialidade de populações de Biomphalaria straminea do Estado de Minas Gerais, como hospedeiras do Schistosoma mansoni Potentiality of the Biomphalaria straminea populations of the State of Minas Gerais, as hosts of Schistosoma mansoni

    Directory of Open Access Journals (Sweden)

    Cecília Pereira de Souza

    1983-09-01

    Full Text Available Caramujos de Biomphalaria straminea, descendentes de exemplares coletados em nove municípios do Estado de Minas Gerais, foram infectados experimentalmente com três cepas de Schistosoma mansoni: "LE", procedente de Belo Horizonte (MG; "SJ", procedente de São José dos Campos (SP e "AL" procedente do Nordeste (AL. As taxas de infeção variaram de 0,0 a 24,0% com a cepa "LE"; de 0,0 a 16% com a cepa "SJ" e de 2,0 a 9,0% com a cepa "AL". Os índices de infecção experimental obtidos foram semelhantes aos registrados por outros autores, para B. straminea dessa região. Comparou-se o número de cercárias de cepa "LE", eliminadas por oito exemplares de B. straminea de Baldim e oito Biomphalaria glabrata do controle, após 30 minutos de exposição à luz. O número de cercárias eliminadas por B. straminea foi de 4.550, aproximadamente cinco vezes menor que o de B. glabrata, 22.679. Discute-se a potencialidade desses moluscos como hospedeiros do S. mansoni nessa região.The decendents of Biomphalaria straminea snails collected in nine regions from the State of Minas Gerais were experimentally infected with three strains of Schistosoma mansoni: "LE", from Belo Horizonte, Minas Gerais; "SJ", from São José dos Campos, State of São Paulo and "AL", from State of Alagoas. The infection rates obtained were of 0 to 24% (LE strain, 0 to 16% (SJ strain and 2 to 9% (AL strain. These infection rates were similar to those obtained by other authors for B. straminea from this region. Comparation were made between the numbers of cercariae (LE strain shed by eight specimens of B. straminea from Baldim and eight B. glabrata of the control group, after 30 minutes of exposure to light. B. straminea shed 4,550 cercariae, about five times less than B. glabrata (22,679. The authors discuss the potentiality of theses molluscs as hosts of S. mansoni in this region.

  12. E-ADA activity in serum of lambs experimentally infected with Haemonchus contortus.

    Science.gov (United States)

    Da Silva, Aleksandro S; Fausto, Guilherme C; Grando, Thirssa H; Cadore, Carlos A; Pimentel, Victor C; Jaques, Jeandre A; Schetinger, Maria R C; Monteiro, Silvia G; Leal, Marta L R

    2013-08-01

    The aim of this study was to evaluate adenosine deaminase (E-ADA) activity in sera of lambs experimentally infected with Haemonchus contortus. We used 12 lambs divided into 2 groups; Group A had 5 healthy, non-infected animals (control) and Group B had 7 healthy animals infected with H. contortus . Lambs were infected orally with 500 larvae (L3) per animal every 2 days, for a period of 20 days, and later the infection was confirmed by examination of feces (eggs per gram [EPG] via fecal egg count). Blood collection was performed at days 0, 20, 40, 60, and 80 post-infection (PI) for analysis of E-ADA activity. Animals in Group A showed negative EPG throughout the experiment unlike those from Group B that had elevated EPG counts. E-ADA activity was reduced in the serum of animals infected with H. contortus when compared to non-infected controls at days 20, 40, 60, and 80 PI. Therefore, it is concluded that infection with H. contortus influences the E-ADA activity in lambs.

  13. Experimental cross-species infection of common marmosets by titi monkey adenovirus.

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    Guixia Yu

    Full Text Available Adenoviruses are DNA viruses that infect a number of vertebrate hosts and are associated with both sporadic and epidemic disease in humans. We previously identified a novel adenovirus, titi monkey adenovirus (TMAdV, as the cause of a fulminant pneumonia outbreak in a colony of titi monkeys (Callicebus cupreus at a national primate center in 2009. Serological evidence of infection by TMAdV was also found in a human researcher at the facility and household family member, raising concerns for potential cross-species transmission of the virus. Here we present experimental evidence of cross-species TMAdV infection in common marmosets (Callithrix jacchus. Nasal inoculation of a cell cultured-adapted TMAdV strain into three marmosets produced an acute, mild respiratory illness characterized by low-grade fever, reduced activity, anorexia, and sneezing. An increase in virus-specific neutralization antibody titers accompanied the development of clinical signs. Although serially collected nasal swabs were positive for TMAdV for at least 8 days, all 3 infected marmosets spontaneously recovered by day 12 post-inoculation, and persistence of the virus in tissues could not be established. Thus, the pathogenesis of experimental inoculation of TMAdV in common marmosets resembled the mild, self-limiting respiratory infection typically seen in immunocompetent human hosts rather than the rapidly progressive, fatal pneumonia observed in 19 of 23 titi monkeys during the prior 2009 outbreak. These findings further establish the potential for adenovirus cross-species transmission and provide the basis for development of a monkey model useful for assessing the zoonotic potential of adenoviruses.

  14. Transcriptomic study of 39 ostreid herpesvirus 1 genes during an experimental infection.

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    Segarra, Amélie; Faury, Nicole; Pépin, Jean-François; Renault, Tristan

    2014-06-01

    Massive mortality outbreaks have been reported in France since 2008 among Pacific oysters, Crassostrea gigas, with the detection of a particular OsHV-1 variant called μVar. Virus infection can be induced in healthy spat in experimental conditions allowing to better understand the disease process, including viral gene expression. Although gene expression of other herpesviruses has been widely studied, we provide the first study following viral gene expression of OsHV-1 over time. In this context, an in vivo transcriptomic study targeting 39 OsHV-1 genes was carried out during an experimental infection of Pacific oyster spat. For the first time, several OsHV-1 mRNAs were detected by real-time PCR at 0 h, 2 h, 4 h, 18 h, 26 h and 42 h post-injection. Several transcripts were detected at 2h post-infection and at 18 h post-infection for all selected ORFs. Quantification of virus gene expression at different times of infection was also carried out using an oyster housekeeping gene, Elongation factor. Developing an OsHV-1-specific reverse transcriptase real time PCR targeting 39 viral gene appears a new tool in terms of diagnosis and can be used to complement viral DNA detection in order to monitor viral replication. Copyright © 2014. Published by Elsevier Inc.

  15. Schistosoma mansoni control in Cul de Sac Valley, Saint Lucia. I. A two-year focal surveillance-mollusciciding programme for the control of Biomphalaria glabrata.

    Science.gov (United States)

    Barnish, G; Christie, J D; Prentice, M A

    1980-01-01

    An area-wide mollusciciding campaign in Cul de Sac valley, St. Lucia reduced incidence of Schistosoma mansoni from 22% to 4.3% between 1970 and 1975. Following this, a two-year focal surveillance-mollusciciding programme was introduced. Sites of potential transmission of S. mansoni were identified and routinely searched for Biomphalaria glabrata. If found, the site was treated with clonitralide 25% emulsifiable concentrate. Two chemotherapy campaigns supplemented the snail control programme. As a result of the combined measures, incidence of the infection dropped from 4.3% to 1.0% and from 2.2% to 0.6% in areas originally of high and low transmission respectively. The cost of protecting the 7,000 population was US $20,362: of these costs, labour absorbed 68%, transport 24%, equipment 4% and molluscicide 4%. The cost per person per year protected was US $1.45 which compares favourably with the $3.24 of the previous scheme. Although effective and relatively cheap, this programme was still dependent on a high standard of supervision for maximum benefit.

  16. Excretion of [3H]prednisolone in clinically normal and experimentally infected bovine udders

    International Nuclear Information System (INIS)

    Geleta, J.N.; Shimoda, W.; Mercer, H.D.

    1984-01-01

    The excretion rate of [3H]prednisolone from clinically normal and experimentally infected udders of 10 lactating cows was studied. Each quarter of 6 cows was injected with a single dose of [3H]prednisolone mixed with non-radioactive prednisolone equivalent to 10 mg in 10 ml of peanut oil base. Each of the remaining 4 cows was given 40 mg of nonradioactive prednisolone and [3H]prednisolone in 60% ethanol IV. Control and postadministration samples of blood, milk, and urine were examined for radioactivity. The effects of [3H]prednisolone were evaluated in the same cows, first in clinically normal udders, then 2 weeks later in udders experimentally infected with Streptococcus agalactiae. Absorption and elimination of prednisolone were the same before and after induced infection. Within 3 hours after intramammary injection, 95% of the labeled prednisolone was absorbed systemically, less than 5% of this dose was recovered in milk, and 29% was excreted in urine. After IV injection of [3H]prednisolone, less than 0.2% of the total radioactivity was recovered in milk and less than 46% was excreted in urine. Clinical mastitis induced by S agalactiae was moderate. Circulating blood leukocytes and somatic cells in the milk of normal cows remained essentially unchanged. The leukocyte response to induced infection was rapid in blood and milk. Large numbers of leukocytes were noticed in the milk and a severe leukopenia occurred. Prednisolone treatment did not alter the number of somatic cells in milk or reduce the inflammatory response of experimentally infected cows

  17. Excretion of (3H)prednisolone in clinically normal and experimentally infected bovine udders

    Energy Technology Data Exchange (ETDEWEB)

    Geleta, J.N.; Shimoda, W.; Mercer, H.D.

    1984-08-01

    The excretion rate of (3H)prednisolone from clinically normal and experimentally infected udders of 10 lactating cows was studied. Each quarter of 6 cows was injected with a single dose of (3H)prednisolone mixed with non-radioactive prednisolone equivalent to 10 mg in 10 ml of peanut oil base. Each of the remaining 4 cows was given 40 mg of nonradioactive prednisolone and (3H)prednisolone in 60% ethanol IV. Control and postadministration samples of blood, milk, and urine were examined for radioactivity. The effects of (3H)prednisolone were evaluated in the same cows, first in clinically normal udders, then 2 weeks later in udders experimentally infected with Streptococcus agalactiae. Absorption and elimination of prednisolone were the same before and after induced infection. Within 3 hours after intramammary injection, 95% of the labeled prednisolone was absorbed systemically, less than 5% of this dose was recovered in milk, and 29% was excreted in urine. After IV injection of (3H)prednisolone, less than 0.2% of the total radioactivity was recovered in milk and less than 46% was excreted in urine. Clinical mastitis induced by S agalactiae was moderate. Circulating blood leukocytes and somatic cells in the milk of normal cows remained essentially unchanged. The leukocyte response to induced infection was rapid in blood and milk. Large numbers of leukocytes were noticed in the milk and a severe leukopenia occurred. Prednisolone treatment did not alter the number of somatic cells in milk or reduce the inflammatory response of experimentally infected cows.

  18. Genetic variability of Taenia solium cysticerci recovered from experimentally infected pigs and from naturally infected pigs using microsatellite markers.

    Science.gov (United States)

    Pajuelo, Mónica J; Eguiluz, María; Roncal, Elisa; Quiñones-García, Stefany; Clipman, Steven J; Calcina, Juan; Gavidia, Cesar M; Sheen, Patricia; Garcia, Hector H; Gilman, Robert H; Gonzalez, Armando E; Zimic, Mirko

    2017-12-01

    The adult Taenia solium, the pork tapeworm, usually lives as a single worm in the small intestine of humans, its only known definitive host. Mechanisms of genetic variation in T. solium are poorly understood. Using three microsatellite markers previously reported [1], this study explored the genetic variability of T. solium from cysts recovered from experimentally infected pigs. It then explored the genetic epidemiology and transmission in naturally infected pigs and adult tapeworms recovered from human carriers from an endemic rural community in Peru. In an initial study on experimental infection, two groups of three piglets were each infected with proglottids from one of two genetically different tapeworms for each of the microsatellites. After 7 weeks, pigs were slaughtered and necropsy performed. Thirty-six (92.3%) out of 39 cysts originated from one tapeworm, and 27 (100%) out of 27 cysts from the other had exactly the same genotype as the parental tapeworm. This suggests that the microsatellite markers may be a useful tool for studying the transmission of T. solium. In the second study, we analyzed the genetic variation of T. solium in cysts recovered from eight naturally infected pigs, and from adult tapeworms recovered from four human carriers; they showed genetic variability. Four pigs had cysts with only one genotype, and four pigs had cysts with two different genotypes, suggesting that multiple infections of genetically distinct parental tapeworms are possible. Six pigs harbored cysts with a genotype corresponding to one of the identified tapeworms from the human carriers. In the dendrogram, cysts appeared to cluster within the corresponding pigs as well as with the geographical origin, but this association was not statistically significant. We conclude that genotyping of microsatellite size polymorphisms is a potentially important tool to trace the spread of infection and pinpoint sources of infection as pigs spread cysts with a shared parental genotype.

  19. Genetic variability of Taenia solium cysticerci recovered from experimentally infected pigs and from naturally infected pigs using microsatellite markers.

    Directory of Open Access Journals (Sweden)

    Mónica J Pajuelo

    2017-12-01

    Full Text Available The adult Taenia solium, the pork tapeworm, usually lives as a single worm in the small intestine of humans, its only known definitive host. Mechanisms of genetic variation in T. solium are poorly understood. Using three microsatellite markers previously reported [1], this study explored the genetic variability of T. solium from cysts recovered from experimentally infected pigs. It then explored the genetic epidemiology and transmission in naturally infected pigs and adult tapeworms recovered from human carriers from an endemic rural community in Peru. In an initial study on experimental infection, two groups of three piglets were each infected with proglottids from one of two genetically different tapeworms for each of the microsatellites. After 7 weeks, pigs were slaughtered and necropsy performed. Thirty-six (92.3% out of 39 cysts originated from one tapeworm, and 27 (100% out of 27 cysts from the other had exactly the same genotype as the parental tapeworm. This suggests that the microsatellite markers may be a useful tool for studying the transmission of T. solium. In the second study, we analyzed the genetic variation of T. solium in cysts recovered from eight naturally infected pigs, and from adult tapeworms recovered from four human carriers; they showed genetic variability. Four pigs had cysts with only one genotype, and four pigs had cysts with two different genotypes, suggesting that multiple infections of genetically distinct parental tapeworms are possible. Six pigs harbored cysts with a genotype corresponding to one of the identified tapeworms from the human carriers. In the dendrogram, cysts appeared to cluster within the corresponding pigs as well as with the geographical origin, but this association was not statistically significant. We conclude that genotyping of microsatellite size polymorphisms is a potentially important tool to trace the spread of infection and pinpoint sources of infection as pigs spread cysts with a shared

  20. PCR diagnostics underestimate the prevalence of avian malaria (Plasmodium relictum) in experimentally-infected passerines

    Science.gov (United States)

    Jarvi, Susan I.; Schultz, Jeffrey J.; Atkinson, Carter T.

    2002-01-01

    Several polymerase chain reaction (PCR)-based methods have recently been developed for diagnosing malarial infections in both birds and reptiles, but a critical evaluation of their sensitivity in experimentally-infected hosts has not been done. This study compares the sensitivity of several PCR-based methods for diagnosing avian malaria (Plasmodium relictum) in captive Hawaiian honeycreepers using microscopy and a recently developed immunoblotting technique. Sequential blood samples were collected over periods of up to 4.4 yr after experimental infection and rechallenge to determine both the duration and detectability of chronic infections. Two new nested PCR approaches for detecting circulating parasites based on P. relictum 18S rRNA genes and the thrombospondin-related anonymous protein (TRAP) gene are described. The blood smear and the PCR tests were less sensitive than serological methods for detecting chronic malarial infections. Individually, none of the diagnostic methods was 100% accurate in detecting subpatent infections, although serological methods were significantly more sensitive (97%) than either nested PCR (61–84%) or microscopy (27%). Circulating parasites in chronically infected birds either disappear completely from circulation or to drop to intensities below detectability by nested PCR. Thus, the use of PCR as a sole means of detection of circulating parasites may significantly underestimate true prevalence.

  1. Hematologic profile of hematophagous Desmodus rotundus bats before and after experimental infection with rabies virus

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    Marilene Fernandes de Almeida

    2014-06-01

    Full Text Available Introduction Hematophagous Desmodus rotundus bats play an important role in the rabies lifecycle. This study describes the hematological profile of these bats before and after experimental infection with rabies virus. Methods Cells counts were performed in a Neubauer chamber. Results The average values of erythrocytes and leucocytes counts in blood before experimental infections were 9.97 × 106mm3 and 4.80 × 103mm3, respectively. Neutrophils represented 69.9% of white blood cells and the lymphocytes represented 26.9%. Following the experimental infections, the average numbers of erythrocytes and leucocytes was 9.43 × 106mm3 and 3.98 × 103mm3, respectively. Neutrophils represented 40% of white blood cells and the lymphocytes represented 59%. Conclusions The hematological profile given in this study can serve as reference values for D. rotundus bats.

  2. Identification of Candida Species Associated with Vulvovaginal Candidiasis by Multiplex PCR

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    Mahnaz Mahmoudi Rad

    2012-01-01

    Full Text Available Background. Vulvovaginal candidiasis is a common infection. The aim of this study was to identify the species of vaginal Candida isolates by using multiplex PCR technique. Methods. 191 isolates from patients admitted to Mahdieh hospital were identified. The vaginal swab specimens were cultured on Sabouraud Dextrose Agar. The ITS1 region between the 18S and 5.8S rRNA genes and a specific DNA fragment within the ITS2 region were amplified. The multiplex PCR products were separated by electrophoresis in 2% agarose gel, visualized by staining with ethidium bromide, and photographed. Descriptive statistics, Chi-square test, and Spearman correlation were used to summarize the findings. Results. C. albicans and C. glabrata were the most common species isolated from the specimens. A mix of C. glabrata and C. albicans was the most common mixed infection isolated from the samples. The analysis revealed a significant positive association between older age and infection with C. glabrata isolates (Spearman’s rho = 0.89, P=0.015. Conclusion. Multiplex PCR is a fast, yet reliable method to identify Candida species. C. albicans and then C. glabrata are the two most common causes of vulvovaginal candidiasis. The number of mixed fungal infections is higher among Iranian population compared to international reports.

  3. Efficacy of albendazole against Taenia multiceps larvae in experimentally infected goats.

    Science.gov (United States)

    Afonso, Sónia M S; Neves, Luis; Pondja, Alberto; Macuamule, Cristiano; Mukaratirwa, Samson; Arboix, Margarita; Cristòfol, Carles; Capece, Bettencourt P S

    2014-12-15

    A controlled trial was conducted to evaluate the efficacy of three therapeutics regimes of albendazole (ABZ) against Taenia multiceps larvae in experimental infected goats. Forty-nine goats experimentally infected with 3000 T. multiceps eggs were selected and randomly divided into treatment or control groups. Treatment with 10mg/kg for 3 days for group 1 (G1), 10mg/kg for group 2 (G2) and 20mg/kg/day for group 3 (G3) was applied 2 months after infection; group 4 (G4) served as a control group. A treatment with doses of 10mg/kg/day for 3 days on group 5 (G5) and group 6 (G6) was used as control, 5 months after the infection. The efficacy of ABZ was assessed as percentage of non-viable cysts which were determined by morphologic characteristics, movement and methyl blue staining technique. The efficacy of ABZ against 2 months old cysts was significantly different from the control and were 90.3% (28/31), 72.7% (8/11) and 73.9% (14/19) for G1, G2 and G3, respectively. No differences were observed in cyst viability between treated and control groups for 5-month old cysts. The results in this study indicate that ABZ is effective in goats against 2-month-old cysts of T. multiceps larva located in tissues outside the brain. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Experimental Infection and Clearance of Coccidian Parasites in Mercury-Exposed Zebra Finches.

    Science.gov (United States)

    Ebers Smith, Jessica H; Cristol, Daniel A; Swaddle, John P

    2018-01-01

    Mercury is a globally distributed, persistent environmental contaminant that affects the health of many taxa. It can suppress the immune system, which often plays a role in defense against parasites. However, there have been few investigations of whether mercury affects the abilities of animals to resist parasitic infection. Here, we exposed zebra finches to a lifetime dietary exposure of methylmercury (1.2 μg/g wet weight) and experimentally infected them with coccidian parasites to examine the effect of methylmercury exposure on parasitic infection. The mercury-exposed birds did not have an altered immune response (heterophil:lymphocyte ratio) nor a reduced ability to clear the infection. However, mercury-exposed birds tended to have higher parasite loads at the time when we expected the greatest immune response (2-3 weeks post-infection). Although mercury did not greatly influence the infection-course of this parasite in captivity, responses may be more accentuated in the wild where birds face additional immune challenges.

  5. Comparison of genome engineering using the CRISPR-Cas9 system in C. glabrata wild-type and lig4 strains.

    Science.gov (United States)

    Cen, Yuke; Timmermans, Bea; Souffriau, Ben; Thevelein, Johan M; Van Dijck, Patrick

    2017-10-01

    Candida glabrata is reported as the second most prevalent human opportunistic fungal pathogen in North America and is threatening patients all over the world. Its incidence is rising, while it has developed resistance to the most widely used antifungal drugs, necessitating new approaches based on better insight into the biology of the organism. Despite its close phylogenetic relationship with Saccharomyces cerevisiae, generating precise genomic alterations in this species is problematic. Previously we have shown that deletion of LIG4, which encodes an enzyme involved in Non-Homologous End Joining (NHEJ), strongly enhances the probability of obtaining correctly modified transformants. In this work we used the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR associated protein 9 (Cas9) system to genetically engineer the C. glabrata genome, targeting the genes ADE2, MET15 and SOK2, located on different chromosomes. We used the CRISPR-Cas9 technology to replace the open reading frame (ORF) by the SAT1 selective marker or introduced a premature stop codon in ADE2 and MET15, as they are easily scored by their adenine or methionine auxotrophy, respectively. The SOK2 gene was modified by insertion of a triple HA-tag sequence and the transformants were verified in a western blot. The CRISPR-Cas9 mediated targeting efficiency varies depending on the gene targeted and the genetic modification performed. We show that CRISPR-Cas9 mediated genome editing is more efficient than the conventional method in the wild-type strain, moreover it has the big advantage being marker-free. In previous work, we showed that the targeting efficiency is highly increased in the lig4Δ strain using the conventional way to delete genes in C. glabrata. Using the CRISPR-Cas9 system in this strain, the percentage of correct transformants is consistently higher compared to the wild-type strain. This indicates that using the lig4 mutant as such is already a strong

  6. Molecular diagnosis of Eimeria stiedae in hepatic tissue of experimentally infected rabbits.

    Science.gov (United States)

    Hassan, Khaled M; Arafa, Waleed M; Mousa, Waheed M; Shokier, Khaled A M; Shany, Salama A; Aboelhadid, Shawky M

    2016-10-01

    The early detection of Eimeria stiedae in the hepatic tissue of experimentally infected rabbits was investigated using molecular assay. Forty 6-week-old male New Zealand rabbits were divided into two groups. Group A (30 animals) was infected with 2.5 × 10(4) sporulated oocysts of E. stiedae per animal on Day 0 and Group B (10 animals) was used as the uninfected controls. Three animals from Group A and one from Group B were sacrificed at 0, 3, 6, 9, 12, 15, 18, 21, 24 and 27 days post infection (PI). Gross and microscopic post-mortem findings were recorded. Polymerase chain reaction (PCR) of the E. stiedae internal transcribed spacer 1 genomic region was conducted on blood, liver tissue, and feces from the Group A experimentally infected animals. Macroscopically, the liver showed irregular yellowish white nodules pathognomonic to E. stiedae infection beginning on Day 15 PI. Hepatomegaly and ascites were obvious from Day 21-24 PI. The presence of different E. stiedae schizonts and gametocytes in the histopathological sections of the biliary epithelium were evident on Day 15 PI. The E. stiedae PCR was first positive in liver tissues on Day 12 and in fecal samples on Day 18 PI, but the blood samples were negative. In conclusion, the PCR can be used for early diagnosis and control of E. stiedae schizonts before shedding of the oocysts in feces. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Influência do magnésio metálico e diferentes sais de magnésio em desovas de Biomphalaria glabrata (Say, 1818 Influence of metallic magnesium and various magnesium salts on egg-masses of Biomphalaria glabrata (Say, 1818

    Directory of Open Access Journals (Sweden)

    Nelymar Martineli Mendes

    1983-12-01

    Full Text Available Soluções de magnésio metálico e diferentes sais de magnésio foram testadas em laboratório, a fim de comprovar a ação das mesmas sobre a oviposição de B. glabrata. Usaram-se, para cada solução, dez caramujos adultos originários da Pampulha, Belo Horizonte-MG (Brasil, criados em laboratório. No experimento I foram estudadas soluções em água desclorada de magnésio elementar (Mg, cloreto de magnésio (MgCl2.6H2O, carbonato de magnésio [(MgCO34.Mg(OH2.nH2O], nitrato de magnésio [Mg(NO32. 6H2O] e sulfato de magnésio (MgSO4.7H2O, e no experimento II, Mg, Mg + latossolo, MgCO3, MgCO3 + latossolo, sempre nas concentrações de 500 e 1.000 ppm. Durante duas semanas os planorbídeos permaneceram em água para adaptação; em seguida, por quatro semanas em exposição com as soluções testadas e, posteriormente, em água por mais duas semanas para recuperação. A cada sete dias as desovas eram contadas, eliminadas e as soluções renovadas. Foi observado que as soluções de Mg e MgCO3, isoladamente ou associada ao latossolo, produziram uma diminuição significativa do número de desovas dos caramujos, mas não parada completa de oviposição, quando comparados os períodos de adaptação com os de exposição. As soluções de Mg e MgCO3 não devem ser consideradas como produtos promissores para o controle da população de B. glabrata, em virtude da diminuição das desovas só terem sido observadas quando foram utilizadas concentrações muito altas.Solutions of magnesium metallic and other salts of magnesium were tested in the laboratory to study the action on B. glabrata oviposition. For each solution, 10 adult snails from Pampulha, Belo Horizonte-MG (Brazil, reared in the laboratory, were used. In experiment I solutions, in dechlorinated water, of magnesium (Mg, magnesium chloride (MgCl2. 6H2O, magnesium carbonate [(MgCO34.Mg(OH2.nH2O], magnesium nitrate [Mg(NO32.6H2O] and magnesium sulfate (MgSO4.7H2O were tested; and in

  8. A paralogue of the phosphomutase-like gene family in Candida glabrata, CgPmu2, gained broad-range phosphatase activity due to a small number of clustered substitutions.

    Science.gov (United States)

    Orlando, Kelly A; Iosue, Christine L; Leone, Sarah G; Davies, Danielle L; Wykoff, Dennis D

    2015-10-15

    Inorganic phosphate is required for a range of cellular processes, such as DNA/RNA synthesis and intracellular signalling. The phosphate starvation-inducible phosphatase activity of Candida glabrata is encoded by the gene CgPMU2 (C. glabrata phosphomutase-like protein). CgPMU2 is part of a three-gene family (∼75% identical) created through gene duplication in the C. glabrata clade; only CgPmu2 is a PHO-regulated broad range acid phosphatase. We identified amino acids that confer broad range phosphatase activity on CgPmu2 by creating fusions of sections of CgPMU2 with CgPMU1, a paralogue with little broad range phosphatase activity. We used site-directed mutagenesis on various fusions to sequentially convert CgPmu1 to CgPmu2. Based on molecular modelling of the Pmu proteins on to a histidine phosphatase crystal structure, clusters of amino acids were found in two distinct regions that were able to confer phosphatase activity. Substitutions in these two regions together conferred broad phosphatase activity on CgPmu1. Interestingly, one change is a histidine adjacent to the active site histidine of CgPmu2 and it exhibits a novel ability to partially replace the conserved active site histidine in CgPmu2. Additionally, a second amino acid change was able to confer nt phosphatase activity to CgPmu1, suggesting single amino acid changes neofunctionalize CgPmu2. © 2015 Authors; published by Portland Press Limited.

  9. Heterogeneous infectiousness in guinea pigs experimentally infected with Trypanosoma cruzi.

    Science.gov (United States)

    Castillo-Neyra, Ricardo; Borrini Mayorí, Katty; Salazar Sánchez, Renzo; Ancca Suarez, Jenny; Xie, Sherrie; Náquira Velarde, Cesar; Levy, Michael Z

    2016-02-01

    Guinea pigs are important reservoirs of Trypanosoma cruzi, the causative parasite of Chagas disease, and in the Southern Cone of South America, transmission is mediated mainly by the vector Triatoma infestans. Interestingly, colonies of Triatoma infestans captured from guinea pig corrals sporadically have infection prevalence rates above 80%. Such high values are not consistent with the relatively short 7-8 week parasitemic period that has been reported for guinea pigs in the literature. We experimentally measured the infectious periods of a group of T. cruzi-infected guinea pigs by performing xenodiagnosis and direct microscopy each week for one year. Another group of infected guinea pigs received only direct microscopy to control for the effect that inoculation by triatomine saliva may have on parasitemia in the host. We observed infectious periods longer than those previously reported in a number of guinea pigs from both the xenodiagnosis and control groups. While some guinea pigs were infectious for a short time, other "super-shedders" were parasitemic up to 22 weeks after infection, and/or positive by xenodiagnosis for a year after infection. This heterogeneity in infectiousness has strong implications for T. cruzi transmission dynamics and control, as super-shedder guinea pigs may play a disproportionate role in pathogen spread. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  10. An experimental model of mycobacterial infection under corneal flaps

    Directory of Open Access Journals (Sweden)

    C.B.D. Adan

    2004-07-01

    Full Text Available In order to develop a new experimental animal model of infection with Mycobacterium chelonae in keratomileusis, we conducted a double-blind prospective study on 24 adult male New Zealand rabbits. One eye of each rabbit was submitted to automatic lamellar keratotomy with the automatic corneal shaper under general anesthesia. Eyes were immunosuppressed by a single local injection of methyl prednisolone. Twelve animals were inoculated into the keratomileusis interface with 1 µl of 10(6 heat-inactivated bacteria (heat-inactivated inoculum controls and 12 with 1 µl of 10(6 live bacteria. Trimethoprim drops (0.1%, w/v were used as prophylaxis for the surgical procedure every 4 h (50 µl, qid. Animals were examined by 2 observers under a slit lamp on the 1st, 3rd, 5th, 7th, 11th, 16th, and 23rd postoperative days. Slit lamp photographs were taken to document clinical signs. Animals were sacrificed when corneal disease was detected and corneal samples were taken for microbiological analysis. Eleven of 12 experimental rabbits developed corneal disease, and M. chelonae could be isolated from nine rabbits. Eleven of the 12 controls receiving a heat-inactivated inoculum did not develop corneal disease. M. chelonae was not isolated from any of the control rabbits receiving a heat-inactivated inoculum, or from the healthy cornea of control rabbits. Corneal infection by M. chelonae was successfully induced in rabbits submitted to keratomileusis. To our knowledge, this is the first animal model of M. chelonae infection following corneal flaps for refractive surgery to be described in the literature and can be used for the analysis of therapeutic responses.

  11. Cnh1 Na+/H+ antiporter and Ena1 Na+-ATPase play different roles in cation homeostasis and cell physiology of Candida glabrata

    Czech Academy of Sciences Publication Activity Database

    Krauke, Yannick; Sychrová, Hana

    2011-01-01

    Roč. 11, č. 1 (2011), s. 29-41 ISSN 1567-1356 R&D Projects: GA MŠk(CZ) LC531 Grant - others:EC(XE) MRTN-CT-2004-512481 Institutional research plan: CEZ:AV0Z50110509 Keywords : Candida glabrata * potassium homeostasis * podium detoxification * Ena 1 ATPase * Cnh1 antiporter Subject RIV: EE - Microbiology, Virology Impact factor: 2.403, year: 2011

  12. Metabolomic profiling in cattle experimentally infected with Mycobacterium avium subsp. paratuberculosis.

    Directory of Open Access Journals (Sweden)

    Jeroen De Buck

    Full Text Available The sensitivity of current diagnostics for Johne's disease, a slow, progressing enteritis in ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP, is too low to reliably detect all infected animals in the subclinical stage. The objective was to identify individual metabolites or metabolite profiles that could be used as biomarkers of early MAP infection in ruminants. In a monthly follow-up for 17 months, calves infected at 2 weeks of age were compared with aged-matched controls. Sera from all animals were analyzed by 1H nuclear magnetic resonance spectrometry. Spectra were acquired, processed, and quantified for analysis. The concentration of many metabolites changed over time in all calves, but some metabolites only changed over time in either infected or non-infected groups and the change in others was impacted by the infection. Hierarchical multivariate statistical analysis achieved best separation between groups between 300 and 400 days after infection. Therefore, a cross-sectional comparison between 1-year-old calves experimentally infected at various ages with either a high- or a low-dose and age-matched non-infected controls was performed. Orthogonal Projection to Latent Structures Discriminant Analysis (OPLS DA yielded distinct separation of non-infected from infected cattle, regardless of dose and time (3, 6, 9 or 12 months after infection. Receiver Operating Curves demonstrated that constructed models were high quality. Increased isobutyrate in the infected cattle was the most important agreement between the longitudinal and cross-sectional analysis. In general, high- and low-dose cattle responded similarly to infection. Differences in acetone, citrate, glycerol and iso-butyrate concentrations indicated energy shortages and increased fat metabolism in infected cattle, whereas changes in urea and several amino acids (AA, including the branched chain AA, indicated increased protein turnover. In conclusion, metabolomics

  13. Antidiarrheal Thymol Derivatives from Ageratina glabrata. Structure and Absolute Configuration of 10-Benzoyloxy-8,9-epoxy-6-hydroxythymol Isobutyrate

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    Celia Bustos-Brito

    2016-09-01

    Full Text Available Chemical investigation of the leaves from Ageratina glabrata yielded four new thymol derivatives, namely: 10-benzoyloxy-8,9-dehydro-6-hydroxythymol isobutyrate (4, 10-benzoyloxy-8,9-dehydrothymol (5, 10-benzoyloxythymol (6 and 10-benzoyloxy-6,8-dihydroxy-9-isobutyryl-oxythymol (7. In addition, (8S-10-benzoyloxy-8,9-epoxy-6-hydroxythymol isobutyrate (1, together with other two already known thymol derivatives identified as 10-benzoyloxy-8,9-epoxy-6-methoxythymol isobutyrate (2 and 10-benzoyloxy-8,9-epoxythymol isobutyrate (3 were also obtained. In this paper, we report the structures and complete assignments of the 1H and 13C-NMR data of compounds 1–7, and the absolute configuration for compound 1, unambiguously established by single crystal X-ray diffraction, and evaluation of the Flack parameter. The in vitro antiprotozoal assay showed that compound 1 and its derivative 1a were the most potent antiamoebic and antigiardial compounds. Both compounds showed selectivity and good antiamoebic activity comparable to emetine and metronidazole, respectively, two antiprotozoal drugs used as positive controls. In relation to anti-propulsive effect, compound 1 and 1a showed inhibitory activity, with activities comparable to quercetin and compound 9, two natural antipropulsive compounds used as positive controls. These data suggest that compound 1 may play an important role in antidiarrheal properties of Ageratina glabrata.

  14. Experimental infection with the small intestinal trematode, Haplorchis pumilio, in young dogs

    DEFF Research Database (Denmark)

    Nissen, Sofie; Nguyen, Lan Anh Thi; Dalsgaard, Anders

    2013-01-01

    included as uninfected controls. Faecal examination for eggs was performed twice weekly using a sieving and sedimentation technique. Body temperature and weight of the dogs were measured as was total white blood cells, blood eosinophils and packed cell volume. Subsets of dogs were examined post......Fishborne zoonotic trematodes (FZT) are highly prevalent in Southeast Asia. Recent studies on the role of domestic animals in the transmission of FZT in Northern Vietnam found that dogs, mainly infected with Haplorchis pumilio, contributed widely to the transmission of FZT. On this background, we...... conducted an experimental infection with H. pumilio to elucidate population dynamics and host reactions in dogs. Eight household-reared dogs (3-6 months old), were each orally infected with 500 H. pumilio metacercariae obtained by artificial digestion of naturally infected fish. Another eight dogs were...

  15. Experimental infection of South American camelids with bluetongue virus serotype 8.

    Science.gov (United States)

    Schulz, Claudia; Eschbaumer, Michael; Rudolf, Miriam; König, Patricia; Keller, Markus; Bauer, Christian; Gauly, Matthias; Grevelding, Christoph G; Beer, Martin; Hoffmann, Bernd

    2012-01-27

    Bluetongue (BT) is an infectious, non-contagious disease of wild and domestic ruminants. It is caused by bluetongue virus (BTV) and transmitted by Culicoides biting midges. Since 1998, BT has been emerging throughout Europe, threatening not only the naïve ruminant population. Historically, South American camelids (SAC) were considered to be resistant to BT disease. However, recent fatalities related to BTV in captive SAC have raised questions about their role in BTV epidemiology. Data on the susceptibility of SAC to experimental infection with BTV serotype 8 (BTV-8) were collected in an animal experiment. Three alpacas (Vicugna pacos) and three llamas (Lama glama) were experimentally infected with BTV-8. They displayed very mild clinical signs. Seroconversion was first measured 6-8 days after infection (dpi) by ELISA, and neutralising antibodies appeared 10-13 dpi. BTV-8 RNA levels in blood were very low, and quickly cleared after seroconversion. However, spleens collected post-mortem were still positive for BTV RNA, over 71 days after the last detection in blood samples. Virus isolation was only possible from blood samples of two alpacas by inoculation of highly sensitive interferon alpha/beta receptor-deficient (IFNAR(-/-)) mice. An in vitro experiment demonstrated that significantly lower amounts of BTV-8 adsorb to SAC blood cells than to bovine blood cells. Although this experiment showed that SAC are generally susceptible to a BTV-8 infection, it indicates that these species play a negligible role in BTV epidemiology. Copyright © 2011 Elsevier B.V. All rights reserved.

  16. Nutritional Status Driving Infection by Trypanosoma cruzi: Lessons from Experimental Animals

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    Guilherme Malafaia

    2011-01-01

    Full Text Available This paper reviews the scientific knowledge about protein-energy and micronutrient malnutrition in the context of Chagas disease, especially in experimental models. The search of articles was conducted using the electronic databases of SciELO (Scientific Electronic Library Online, PubMed and MEDLINE published between 1960 and March 2010. It was possible to verify that nutritional deficiencies (protein-energy malnutrition and micronutrient malnutrition exert a direct effect on the infection by T. cruzi. However, little is known about the immunological mechanisms involved in the relationship “nutritional deficiencies and infection by T. cruzi”. A hundred years after the discovery of Chagas disease many aspects of this illness still require clarification, including the effects of nutritional deficiencies on immune and pathological mechanisms of T. cruzi infection.

  17. Experimental infection of Rickettsia parkeri in the Rhipicephalus microplus tick.

    Science.gov (United States)

    Cordeiro, Matheus Dias; de Azevedo Baêta, Bruna; Cepeda, Patricia Barizon; Teixeira, Rafaella Câmara; Ribeiro, Carla Carolina Dias Uzedo; de Almeida Valim, Jaqueline Rodrigues; Pinter, Adriano; da Fonseca, Adivaldo Henrique

    2018-01-01

    This study aimed to evaluate, by means of artificial feeding, the interaction between a pathogenic rickettsia and the hard tick R. microplus. We used partially engorged females fed on calves free of Rickettsia spp. Group 1 (G1), containing 20 ticks, was fed bovine blood only. Group 2 (G2), containing 20 ticks, was fed blood containing uninfected VERO cells, and group 3 (G3), containing 40 ticks, was fed blood containing VERO cells infected with Rickettsia parkeri. Biological parameters of the non-parasitic phase and a possible bacterial transmission to the tick eggs and to guinea pigs were evaluated. At the end of oviposition, all G3 females were PCR-positive for genes specific for the genus Rickettsia. Although no guinea pigs were infected, the experimental infection of R. microplus by R. parkeri caused a deleterious effect on the oviposition and provided the first report of transovarian transmission of rickettsia in this tick. Copyright © 2017 Elsevier GmbH. All rights reserved.

  18. Experimental Infection with Sporulated Oocysts of Eimeria maxima (Apicomplexa: Eimeriidae) in Broiler

    Science.gov (United States)

    Brito, Luciana da S.; Pereira, Elder N.; da Silva, Augusta A.; Bentivóglio Costa Silva, Vinícius; Freitas, Fagner L. da C.

    2014-01-01

    Through this study we assessed the metabolic and pathological changes in broilers experimentally infected with oocysts of Eimeria maxima. To perform the experiment, we used 150 broiler strain cooB males, with ten days of age, were randomized according to weight and randomly assigned to two experimental groups: the control group was inoculated with 0.5 mL of distilled water; the infected group inoculated with 0.5 mL of solution containing 5 × 104 sporulated oocysts of Eimeria maxima. The live performance was evaluated on day 0 (day of inoculation), 5°, 10°, 15°, 25°, and 35° dpi, being slaughtered by cervical dislocation, fifteen birds/group. Although the sum in meat production was higher in the control group, the weight of the heart and gizzard of the experimental animals showed no significant difference, while the liver had difference on day 5°, 15°, and 35° dpi. The pathologic evaluation showed congested mucosa and presence of large amounts of mucus at 6 dpi. Therefore, it is concluded that the dose of 5 × 104  E. maxima inoculated in the experimental group was enough to cause harm to the animal organism. PMID:26464925

  19. Effect of some parasitic infection on neurotransmitters in the brain of experimentally infected mice before and after treatment.

    Science.gov (United States)

    Abdel Ghafar, A E; Elkowrany, S E; Salem, S A; Menaisy, A A; Fadel, W A; Awara, W M

    1996-08-01

    The effects of some parasitic infection (bilharziasis, toxocariasis and trichinosis) on the brain of experimentally infected mice were investigated. Eighty animals were classified into four groups, group I contained five non infected animals as a control group. The other groups each contained twenty-five mice infected with Schistosoma mansoni (group II), Toxocara canis (group III) and Trichinella spiralis (group IV). Each infected group was divided into two subgroups (a,b). Subgroup (a) left untreated and subgroups (b) treated by praziquantel (in group II) and mebendazole (in group III and IV). Histopathological and immunological examination using peroxidase antiperoxidase (PAP) technique and neurotransmitters estimation (nor-epinephrine, dopamine and serotonine) were carried. In the untreated animals, there were mild histopathological changes and mild antigenic deposition in subgroups (IIa and IIIa) and marked changes in subgroup (IVa). There were significant decrease in dopamine in subgroup (IIIa), not improved after treatment (subgroup IIIb) and significant decrease in nor-epinephrine and serotonine in subgroup (IVa) improved after treatment in subgroup (IVb). The neurotransmitters changes may explain the motor, behavioural and emotional changes that occurred with these parasites.

  20. Aspectos da ecologia reprodutiva de Pera glabrata (Schott Poepp. ex Baill. (Euphorbiaceae em uma área de Cerrado no estado de São Paulo

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    Juliana Ribeirão de Freitas

    2011-12-01

    Full Text Available Pera glabrata é uma árvore que apresenta ampla distribuição no Brasil. A espécie vegetal é de importância para a conservação e recuperação de áreas degradadas, pois está presente em áreas impactadas, produz e dispersa grande quantidade de sementes e constitui-se em fonte alimentar para elevado número de espécies animais. Apesar da importância fitossociológica da espécie, ainda não existem estudos que abordem a sua ecologia reprodutiva. Este trabalho teve como objetivo caracterizar aspectos da fenologia reprodutiva, da morfologia floral, dos sistemas reprodutivo e de polinização e da dispersão de sementes da espécie. O estudo foi realizado em uma área de Cerrado no município de São Carlos, SP. Verificou-se que Pera glabrata é dioica e apresenta floração massiva e as flores dos dois sexos são pequenas, involucradas, amarelas e de antese diurna. As flores masculinas apresentam néctar e emitem odor adocicado, e as femininas não oferecem recursos perceptíveis aos visitantes florais. As flores foram visitadas por 32 espécies de Diptera e Hymenoptera de pequeno porte. Ocorre a formação de frutos e sementes por autogamia. Foram identificadas 25 espécies de aves visitando indivíduos com frutos maduros, das quais 16 ingeriram as sementes ariladas. Pera glabrata é autogâmica, com síndrome de polinização por diversos pequenos insetos e com dispersão ornitocórica de suas sementes.

  1. Experimental models of acute infection and Toll-like receptor driven septic shock.

    Science.gov (United States)

    Ferstl, Ruth; Spiller, Stephan; Fichte, Sylvia; Dreher, Stefan; Kirschning, Carsten J

    2009-01-01

    Mainly Gram-negative and Gram-positive bacterial infections, but also other infections such as with fungal or viral pathogens, can cause the life-threatening clinical condition of septic shock. Transgression of the host immune response from a local level limited to the pathogen's place of entry to the systemic level is recognised as a major mode of action leading to sepsis. This view has been established upon demonstration of the capacity of specific pathogen-associated molecular patterns (PAMPs) to elicit symptoms of septic shock upon systemic administration. Immune stimulatory PAMPs are agonists of soluble, cytoplasmic, as well as/or cell membrane-anchored and/or -spanning pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs). However, reflection of pathogen-host crosstalk triggering sepsis pathogenesis upon an infection by a host response to challenge with an isolated PAMP is incomplete. Therefore, an experimental model more reflective of pathogen-host interaction requires experimental host confrontation with a specific pathogen in its viable form resulting in a collective stimulation of a variety of specific PRRs. This chapter describes methods to analyse innate pathogen sensing by the host on both a cellular and systemic level.

  2. Technetium-99m stannous pyrophosphate imaging of experimental infective endocarditis

    International Nuclear Information System (INIS)

    Riba, A.L.; Downs, J.; Thakur, M.L.; Gottschalk, A.; Andriole, V.T.; Zaret, B.L.

    1978-01-01

    Technetium-99m stannous pyrophosphate (/sup 99m/Tc-PYP) cardiac scintigraphy was performed in 15 rabbits with experimental Streptococcus sanguis aortic-valve infective endocarditis. The animals were imaged five to seven days after the administration of bacteria, and in each case abnormal accumulation of the tracer was visualized in the region of the aortic valve. Three types of cardiac scintigraphic patterns were demonstrated: focal, multifocal, and extensive, each correlating well with the anatomical extent of the lesion as defined by gross pathology. Tissue distribution studies demonstrated a 30 +- 5.3 (mean +- SEM) fold excess of radionuclide uptake in the infective endocarditis lesion compared with that of normal myocardium. Imaging of excised hearts from four animals showed an excellent correlation with in vivo imaging as well as gross pathology. Five animals with nonbacterial thrombotic aortic valve endocarditis demonstrated similar scintigraphic and tissue distribution results. In contrast, four normal animals failed to demonstrate abnormal /sup 99m/Tc-PYP cardiac scintigrams or tissue uptake. This study demonstrates that /sup 99m/Tc-PYP cardiac scintigraphy is a sensitive technique to detect experimental aortic valve endocarditis

  3. Horses experimentally infected with Sarcocystis neurona develop altered immune responses in vitro.

    Science.gov (United States)

    Witonsky, Sharon G; Ellison, Siobhan; Yang, Jibing; Gogal, Robert M; Lawler, Heather; Suzuki, Yasuhiro; Sriranganathan, Namalwar; Andrews, Frank; Ward, Daniel; Lindsay, David S

    2008-10-01

    Equine protozoal myeloencephalitis (EPM) due to Sarcocystis neurona infection is 1 of the most common neurologic diseases in horses in the United States. The mechanisms by which most horses resist disease, as well as the possible mechanisms by which the immune system may be suppressed in horses that develop EPM, are not known. Therefore, the objectives of this study were to determine whether horses experimentally infected with S. neurona developed suppressed immune responses. Thirteen horses that were negative for S. neurona antibodies in serum and cerebrospinal fluid (CSF) were randomly assigned to control (n = 5) or infected (n = 8) treatment groups. Neurologic exams and cerebrospinal fluid analyses were performed prior to, and following, S. neurona infection. Prior to, and at multiple time points following infection, immune parameters were determined. All 8 S. neurona-infected horses developed clinical signs consistent with EPM, and had S. neurona antibodies in the serum and CSF. Both infected and control horses had increased percentages (P < 0.05) of B cells at 28 days postinfection. Infected horses had significantly decreased (P < 0.05) proliferation responses as measured by thymidine incorporation to nonspecific mitogens phorbol myristate acetate (PMA) and ionomycin (I) as soon as 2 days postinfection.

  4. Experimental infection of cattle, sheep and pigs with 'Hobi'-like pestivirus.

    Science.gov (United States)

    Decaro, Nicola; Mari, Viviana; Lucente, Maria Stella; Sciarretta, Rossana; Moreno, Ana; Armenise, Carlo; Losurdo, Michele; Camero, Michele; Lorusso, Eleonora; Cordioli, Paolo; Buonavoglia, Canio

    2012-03-23

    To date, limited information is available on the ability of 'Hobi'-like pestiviruses (putative bovine viral diarrhoea 3) to infect and cause disease in animal species traditionally affected by pestiviruses. In order to obtain new insights into host range and pathogenic potential of this atypical pestivirus, BVDV-seronegative calves (n=5), lambs (n=5) and piglets (n=5) were experimentally infected with the European 'Hobi'-like strain Italy-1/10-1, whereas two animals per species served as uninfected controls. Appearance of clinical signs, leukopenia, viremia, viral shedding and seroconversion were monitored for 28 days post-infection. Calves and lambs were successfully infected, displaying respiratory signs (nasal discharge), moderate hyperthermia and leukopenia, viremia and viral shedding through the nasal and faecal routes. Antibody responses were observed in both animal species by ELISA and virus neutralisation assays. In contrast, inoculated piglets did not display any clinical signs nor leukopenia and viral RNA was not detected in any biological samples. Nevertheless, the presence of detectable antibodies by virus neutralisation accounted for a successful, albeit limited infection of these animals. Copyright © 2011 Elsevier B.V. All rights reserved.

  5. Experimental feline enteric coronavirus infection reveals an aberrant infection pattern and shedding of mutants with impaired infectivity in enterocyte cultures

    Science.gov (United States)

    Desmarets, Lowiese M. B.; Vermeulen, Ben L.; Theuns, Sebastiaan; Conceição-Neto, Nádia; Zeller, Mark; Roukaerts, Inge D. M.; Acar, Delphine D.; Olyslaegers, Dominique A. J.; Van Ranst, Marc; Matthijnssens, Jelle; Nauwynck, Hans J.

    2016-01-01

    Feline infectious peritonitis (FIP) results from mutations in the viral genome during a common feline enteric coronavirus (FECV) infection. Since many virological and immunological data on FECV infections are lacking, the present study investigated these missing links during experimental infection of three SPF cats with FECV strain UCD. Two cats showed mild clinical signs, faecal shedding of infectious virus from 4 dpi, a cell-associated viraemia at inconsistent time points from 5 dpi, a highly neutralising antibody response from 9 dpi, and no major abnormalities in leukocyte numbers. Faecal shedding lasted for 28–56 days, but virus shed during this stage was less infectious in enterocyte cultures and affected by mutations. Remarkably, in the other cat neither clinical signs nor acute shedding were seen, but virus was detected in blood cells from 3 dpi, and shedding of non-enterotropic, mutated viruses suddenly occurred from 14 dpi onwards. Neutralising antibodies arose from 21 dpi. Leukocyte numbers were not different compared to the other cats, except for the CD8+ regulatory T cells. These data indicate that FECV can infect immune cells even in the absence of intestinal replication and raise the hypothesis that the gradual adaptation to these cells can allow non-enterotropic mutants to arise. PMID:26822958

  6. Experimental and natural infections in MyD88- and IRAK-4-deficient mice and humans

    Science.gov (United States)

    von Bernuth, Horst; Picard, Capucine; Puel, Anne; Casanova, Jean-Laurent

    2013-01-01

    Most Toll-like-receptors (TLRs) and interleukin-1 receptors (IL-1Rs) signal via myeloid differentiation primary response 88 (MyD88) and interleukin-1 receptor-associated kinase 4 (IRAK-4). The combined roles of these two receptor families in the course of experimental infections have been assessed in MyD88- and IRAK-4-deficient mice for almost fifteen years. These animals have been shown to be susceptible to 46 pathogens: 27 bacteria, 8 viruses, 7 parasites, and 4 fungi. Humans with inborn MyD88 or IRAK-4 deficiency were first identified in 2003. They suffer from naturally occurring life-threatening infections caused by a small number of bacterial species, although the incidence and severity of these infections decrease with age. Mouse TLR- and IL-1R-dependent immunity mediated by MyD88 and IRAK-4 seems to be vital to combat a wide array of experimentally administered pathogens at most ages. By contrast, human TLR- and IL-1R-dependent immunity mediated by MyD88 and IRAK-4 seems to be effective in the natural setting against only a few bacteria and is most important in infancy and early childhood. The roles of TLRs and IL-1Rs in protective immunity deduced from studies in mutant mice subjected to experimental infections should therefore be reconsidered in the light of findings for natural infections in humans carrying mutations as discussed in this review. PMID:23255009

  7. Chimeras between C. glabrata Cnh1 and S. cerevisiae Nha1 Naplus/Hplus-antiporters are functional proteins increasing the salt tolerance of yeast cells

    Czech Academy of Sciences Publication Activity Database

    Krauke, Yannick; Sychrová, Hana

    2010-01-01

    Roč. 55, č. 5 (2010), s. 435-441 ISSN 0015-5632 R&D Projects: GA MŠk(CZ) LC531 Grant - others:EC(XE) MRTN-CT-2004-512481 Institutional research plan: CEZ:AV0Z50110509 Keywords : C. glabrata * S. cerevisiae * Nha1 antiporter Subject RIV: EE - Microbiology, Virology Impact factor: 0.977, year: 2010

  8. Endogenous growth factor stimulation of hemocyte proliferation induces resistance to Schistosoma mansoni challenge in the snail host.

    Science.gov (United States)

    Pila, Emmanuel A; Gordy, Michelle A; Phillips, Valerie K; Kabore, Alethe L; Rudko, Sydney P; Hanington, Patrick C

    2016-05-10

    Digenean trematodes are a large, complex group of parasitic flatworms that infect an incredible diversity of organisms, including humans. Larval development of most digeneans takes place within a snail (Gastropoda). Compatibility between snails and digeneans is often very specific, such that suitable snail hosts define the geographical ranges of diseases caused by these worms. The immune cells (hemocytes) of a snail are sentinels that act as a crucial barrier to infection by larval digeneans. Hemocytes coordinate a robust and specific immunological response, participating directly in parasite killing by encapsulating and clearing the infection. Hemocyte proliferation and differentiation are influenced by unknown digenean-specific exogenous factors. However, we know nothing about the endogenous control of hemocyte development in any gastropod model. Here, we identify and functionally characterize a progranulin [Biomphalaria glabrata granulin (BgGRN)] from the snail B. glabrata, a natural host for the human blood fluke Schistosoma mansoni Granulins are growth factors that drive proliferation of immune cells in organisms, spanning the animal kingdom. We demonstrate that BgGRN induces proliferation of B. glabrata hemocytes, and specifically drives the production of an adherent hemocyte subset that participates centrally in the anti-digenean defense response. Additionally, we demonstrate that susceptible B. glabrata snails can be made resistant to infection with S. mansoni by first inducing hemocyte proliferation with BgGRN. This marks the functional characterization of an endogenous growth factor of a gastropod mollusc, and provides direct evidence of gain of resistance in a snail-digenean infection model using a defined factor to induce snail resistance to infection.

  9. Anatomopathological study in BALB/c mice brains experimentally infected with Toxoplasma gondii

    Directory of Open Access Journals (Sweden)

    Marcos Gontijo da Silva

    Full Text Available Toxoplasmosis is one of the most important diseases of the nervous central system, leading to severe symptoms and, many times, irreversible sequelae. This work demonstrated the main anatomopathological lesions caused by Toxoplasma gondii in brains from experimentally infected BALB/c mice. We analyzed 51 cases of mice that developed toxoplasmosis after experimental infection by intraperitoneal inoculation of blood, amniotic liquid and cerebrospinal fluid from fetuses, newly born children and pregnant women with clinical and laboratory signals of toxoplasmosis. In all experiments where we detected the parasite in mice we also detected pathological lesions in the animal brains with great polymorphism between experiments. Edema was the most found lesion in all cases. Besides, it was possible to demonstrate the inflammatory process in 82.4% of cases and necrosis in 64.7% of cases, in agreement with the literature that describes severe neurological damage in its hosts.

  10. Experimental infection of serotine bats (Eptesicus serotinus) with European bat lyssavirus type 1a.

    Science.gov (United States)

    Freuling, C; Vos, A; Johnson, N; Kaipf, I; Denzinger, A; Neubert, L; Mansfield, K; Hicks, D; Nuñez, A; Tordo, N; Rupprecht, C E; Fooks, A R; Müller, T

    2009-10-01

    The serotine bat (Eptesicus serotinus) accounts for the vast majority of bat rabies cases in Europe and is considered the main reservoir for European bat lyssavirus type 1 (EBLV-1, genotype 5). However, so far the disease has not been investigated in its native host under experimental conditions. To assess viral virulence, dissemination and probable means of transmission, captive bats were infected experimentally with an EBLV-1a virus isolated from a naturally infected conspecific from Germany. Twenty-nine wild caught bats were divided into five groups and inoculated by intracranial (i.c.), intramuscular (i.m.) or subcutaneous (s.c.) injection or by intranasal (i.n.) inoculation to mimic the various potential routes of infection. One group of bats was maintained as uninfected controls. Mortality was highest in the i.c.-infected animals, followed by the s.c. and i.m. groups. Incubation periods varied from 7 to 26 days depending on the route of infection. Rabies did not develop in the i.n. group or in the negative-control group. None of the infected bats seroconverted. Viral antigen was detected in more than 50% of the taste buds of an i.c.-infected animal. Shedding of viable virus was measured by virus isolation in cell culture for one bat from the s.c. group at 13 and 14 days post-inoculation, i.e. 7 days before death. In conclusion, it is postulated that s.c. inoculation, in nature caused by bites, may be an efficient way of transmitting EBLV-1 among free-living serotine bats.

  11. Lipid and glucose metabolism of broilers (Gallus gallus domesticus experimentally infected with Eimeria acervulina Tyzzer, 1929 oocysts

    Directory of Open Access Journals (Sweden)

    FLC Freitas

    2008-09-01

    Full Text Available Lipid and glucose metabolism of 76 ten-day-old Cobb male broilers, experimentally infected with Eimeria acervulina, was studied for 30 days. Birds were distributed in 2 groups: one infected with 1x10(6 E. acervulina sporulated oocysts, and the other inoculated with distilled water. Pathological e biochemical liver changes were assessed, as well as plasma glucose concentrations and total cholesterol, HDL, LDL, VLDL, fatty-acid, and triglyceride levels in the serum. The infected broilers presented hypoglycemia associated with a reduction in liver glycogen. In addition, these birds developed fatty liver, and there were changes in all lipid classes in the serum. Lipid and glucose metabolism was dramatically changed in broilers experimentally infected with 1x10(6 E. acervulina oocysts.

  12. Experimental Andes virus infection in deer mice: characteristics of infection and clearance in a heterologous rodent host.

    Directory of Open Access Journals (Sweden)

    Jessica R Spengler

    Full Text Available New World hantaviruses can cause hantavirus cardiopulmonary syndrome with high mortality in humans. Distinct virus species are hosted by specific rodent reservoirs, which also serve as the vectors. Although regional spillover has been documented, it is unknown whether rodent reservoirs are competent for infection by hantaviruses that are geographically separated, and known to have related, but distinct rodent reservoir hosts. We show that Andes virus (ANDV of South America, carried by the long tailed pygmy rice rat (Oligoryzomys longicaudatus, infects and replicates in vitro and in vivo in the deer mouse (Peromyscus maniculatus, the reservoir host of Sin Nombre virus (SNV, found in North America. In experimentally infected deer mice, viral RNA was detected in the blood, lung, heart and spleen, but virus was cleared by 56 days post inoculation (dpi. All of the inoculated deer mice mounted a humoral immune response by 14 dpi, and produced measurable amounts of neutralizing antibodies by 21 dpi. An up-regulation of Ccl3, Ccl4, Ccl5, and Tgfb, a strong CD4⁺ T-cell response, and down-regulation of Il17, Il21 and Il23 occurred during infection. Infection was transient with an absence of clinical signs or histopathological changes. This is the first evidence that ANDV asymptomatically infects, and is immunogenic in deer mice, a non-natural host species of ANDV. Comparing the immune response in this model to that of the immune response in the natural hosts upon infection with their co-adapted hantaviruses may help clarify the mechanisms governing persistent infection in the natural hosts of hantaviruses.

  13. Laboratory experimental infection of sheep to Ornithobilharzia turkestanicum and its confirmation using post-mortem examination and histopathology

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    gholamreza karimi

    2014-11-01

    Full Text Available Ornithobilharzia turkestanicum from genus Ornithobilharzia genus and family Schistosomatidae is an important agent of parasitological infection in sheep. This parasite has been reported from Russia, China, Turkestan (Kazakhstan, Kyrgyzstan, Turkmenistan and Uzbekistan, Pakistan, Iraq, Turkey and Iran. Parasitological infection due to this agent could be one of the important factors of decreasing the production rate of livestock in Iran. The purpose of this study, was to experimentally infect sheep with this parasite and confirm the infection by post-mortem examination and Histopathology which was done successfully. Twenty five sheep were used in the study of which 10 sheep were experimentally infected by Ornithobilharzia turkestanikum using subcutaneous injection and 10 sheep by skin contact method and the other 5 sheep were kept as control. Result of post-mortem and Histopathology during a one year period confirmed that all of sheep were infected and adult worm, were seen in their mesentery. Mean number of cercaria used for inducing the infection was 6425 and 462 adult worms were collected post-mortem. There was no significant relationship between the number of cercaria and adult worms collected. Male sheep were more infected than female.

  14. Carbohydrate-rich high-molecular-mass antigens are strongly recognized during experimental Histoplasma capsulatum infection

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    Fabrine Sales Massafera Tristão

    2012-04-01

    Full Text Available INTRODUCTION: During histoplasmosis, Histoplasma capsulatum soluble antigens (CFAg can be naturally released by yeast cells. Because CFAg can be specifically targeted during infection, in the present study we investigated CFAg release in experimental murine histoplasmosis, and evaluated the host humoral immune response against high-molecular-mass antigens (hMMAg. >150 kDa, the more immunogenic CFAg fraction. METHODS: Mice were infected with 2.2x10(4 H. capsulatum IMT/HC128 yeast cells. The soluble CFAg, IgG anti-CFAg, IgG anti-hMMAg, and IgG-hMMAg circulating immune complexes (CIC levels were determined by enzymelinked immunosorbent assay, at days 0, 7, 14, and 28 post-infection. RESULTS: We observed a progressive increase in circulating levels of CFAg, IgG anti-CFAg, IgG anti-hMMAg, and IgG-hMMAg CIC after H. capsulatum infection. The hMMAg showed a high percentage of carbohydrates and at least two main immunogenic components. CONCLUSIONS: We verified for the first time that hMMAg from H. capsulatum IMT/HC128 strain induce humoral immune response and lead to CIC formation during experimental histoplasmosis.

  15. Persistence of experimental Rocio virus infection in the golden hamster (Mesocricetus auratus

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    Daniele Freitas Henriques

    2012-08-01

    Full Text Available Rocio virus (ROCV is an encephalitic flavivirus endemic to Brazil. Experimental flavivirus infections have previously demonstrated a persistent infection and, in this study, we investigated the persistence of ROCV infection in golden hamsters (Mesocricetus auratus. The hamsters were infected intraperitoneally with 9.8 LD50/0.02 mL of ROCV and later anaesthetised and sacrificed at various time points over a 120-day period to collect of blood, urine and organ samples. The viral titres were quantified by real-time-polymerase chain reaction (qRT-PCR. The specimens were used to infect Vero cells and ROCV antigens in the cells were detected by immunefluorescence assay. The levels of antibodies were determined by the haemagglutination inhibition technique. A histopathological examination was performed on the tissues by staining with haematoxylin-eosin and detecting viral antigens by immunohistochemistry (IHC. ROCV induced a strong immune response and was pathogenic in hamsters through neuroinvasion. ROCV was recovered from Vero cells exposed to samples from the viscera, brain, blood, serum and urine and was detected by qRT-PCR in the brain, liver and blood for three months after infection. ROCV induced histopathological changes and the expression of viral antigens, which were detected by IHC in the liver, kidney, lung and brain up to four months after infection. These findings show that ROCV is pathogenic to golden hamsters and has the capacity to cause persistent infection in animals after intraperitoneal infection.

  16. Experimental infection with Brazilian Newcastle disease virus strain in pigeons and chickens

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    Adriano de Oliveira Torres Carrasco

    2016-03-01

    Full Text Available Abstract This study was designed with the goal of adding as much information as possible about the role of pigeons (Columba livia and chickens (Gallus gallus in Newcastle disease virus epidemiology. These species were submitted to direct experimental infection with Newcastle disease virus to evaluate interspecies transmission and virus-host relationships. The results obtained in four experimental models were analyzed by hemagglutination inhibition and reverse transcriptase polymerase chain reaction for detection of virus shedding. These techniques revealed that both avian species, when previously immunized with a low pathogenic Newcastle disease virus strain (LaSota, developed high antibody titers that significantly reduced virus shedding after infection with a highly pathogenic Newcastle disease virus strain (São Joao do Meriti and that, in chickens, prevent clinical signs. Infected pigeons shed the pathogenic strain, which was not detected in sentinel chickens or control birds. When the presence of Newcastle disease virus was analyzed in tissue samples by RT-PCR, in both species, the virus was most frequently found in the spleen. The vaccination regimen can prevent clinical disease in chickens and reduce viral shedding by chickens or pigeons. Biosecurity measures associated with vaccination programs are crucial to maintain a virulent Newcastle disease virus-free status in industrial poultry in Brazil.

  17. Pathogenesis of Riemerella anatipestifer in turkeys after experimental mono-infection via respiratory routes or dual infection together with the avian metapneumovirus.

    Science.gov (United States)

    Rubbenstroth, Dennis; Ryll, Martin; Behr, Klaus-Peter; Rautenschlein, Silke

    2009-12-01

    Riemerella anatipestifer (RA) is the causative agent of septicaemic and exudative diseases in a variety of bird species. Despite numerous outbreaks, little is known about the pathogenicity of RA for turkeys. We investigated the development of RA-induced disease in commercial turkey poults following RA inoculation via different respiratory routes. Inoculation by aerosol or injection into the abdominal air sac led to systemic infection and mild gross lesions, including pericarditis, epicarditis and airsacculitis, which were less pronounced compared with field outbreaks. It was speculated, that viral pathogens, such as the avian metapneumovirus (aMPV), may exacerbate RA pathogenesis under field conditions. We inoculated turkey poults with virulent aMPV. Subsequently, aMPV-infected and virus-free birds were exposed 3 to 5 days later to a high dose of RA by aerosol (>10(10) colony-forming units/ml in 8 ml aerosol per 11 or 12 birds) or were inoculated 4 days later with a low RA dose (10(4.9) colony-forming units per bird) via the intranasal route. Intranasal RA inoculation with the low bacterial dose led to a respiratory and systemic RA infection in aMPV-infected birds, while virus-free birds remained RA-negative. Following exposure to a high RA dose by aerosol, aMPV-infected groups showed slightly enhanced incidences of gross lesions and RA re-isolation. The present study clearly confirms that RA is pathogenic for turkeys after experimental inoculation via respiratory routes, which are speculated to be the natural route of infection. However, experimental models in this study did not reproduce the severity of RA-related disease as observed under field conditions, which emphasizes the importance of other contributing factors. aMPV-induced respiratory lesions may serve as a predisposing factor for the establishment of RA infection, since they favour colonization of the bacterium.

  18. Facultative Sterol Uptake in an Ergosterol-Deficient Clinical Isolate of Candida glabrata Harboring a Missense Mutation in ERG11 and Exhibiting Cross-Resistance to Azoles and Amphotericin B

    NARCIS (Netherlands)

    Hull, Claire M.; Parker, Josie E.; Bader, Oliver; Weig, Michael; Gross, Uwe; Warrilow, Andrew G. S.; Kelly, Diane E.; Kelly, Steven L.

    We identified a clinical isolate of Candida glabrata (CG156) exhibiting flocculent growth and cross-resistance to fluconazole (FLC), voriconazole (VRC), and amphotericin B (AMB), with MICs of >256, >256, and 32 mu g ml(-1), respectively. Sterol analysis using gas chromatography-mass spectrometry

  19. Morphology-Independent Virulence of Candida Species during Polymicrobial Intra-abdominal Infections with Staphylococcus aureus.

    Science.gov (United States)

    Nash, Evelyn E; Peters, Brian M; Fidel, Paul L; Noverr, Mairi C

    2016-01-01

    Intra-abdominal polymicrobial infections cause significant morbidity and mortality. An experimental mouse model of Candida albicans-Staphylococcus aureus intra-abdominal infection (IAI) results in 100% mortality by 48 to 72 h postinoculation, while monomicrobial infections are avirulent. Mortality is associated with robust local and systemic inflammation without a requirement for C. albicans morphogenesis. However, the contribution of virulence factors coregulated during the yeast-to-hypha transition is unknown. This also raised the question of whether other Candida species that are unable to form hyphae are as virulent as C. albicans during polymicrobial IAI. Therefore, the purpose of this study was to evaluate the ability of non-albicans Candida (NAC) species with various morphologies and C. albicans transcription factor mutants (efg1/efg1 and cph1/cph1) to induce synergistic mortality and the accompanying inflammation. Results showed that S. aureus coinoculated with C. krusei or C. tropicalis was highly lethal, similar to C. albicans, while S. aureus-C. dubliniensis, S. aureus-C. parapsilosis, and S. aureus-C. glabrata coinoculations resulted in little to no mortality. Local and systemic interleukin-6 (IL-6) and prostaglandin E2 (PGE2) levels were significantly elevated during symptomatic and/or lethal coinfections, and hypothermia strongly correlated with mortality. Coinoculation with C. albicans strains deficient in the transcription factor Efg1 but not Cph1 reversed the lethal outcome. These results support previous findings and demonstrate that select Candida species, without reference to any morphological requirement, induce synergistic mortality, with IL-6 and PGE2 acting as key inflammatory factors. Mechanistically, signaling pathways controlled by Efg1 are critical for the ability of C. albicans to induce mortality from an intra-abdominal polymicrobial infection. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  20. Coronative antibody tires in sera of healthy adults and experimentally infected volunteers.

    Science.gov (United States)

    Bradburne, A F; Somerset, B A

    1972-06-01

    Six coronaviruses isolated in the U.S.A. have been inoculated into volunteers and all produced colds. Between 10 and 20% of infected volunteers developed heterologous antibody responses after these and other experimental infections with coronaviruses. The haemagglutination-inhibition test with the OC43 virus strain was found to detect antibody rises after infection with a variety of strains.Studies on normal adult sera taken between 1965 and 1970 revealed a high frequency of neutralizing antibody to one strain (229 E) and a frequency of HI antibody to strain OC43 which fluctuated from year to year. Complement-fixing antibodies to these two viruses were also found, revealing an apparent increase in the activity of coronaviruses in the general population of the U.K., during the winter of 1968-9.

  1. Experimental gonococcal infection in male volunteers: Cumulative experience with Neisseria gonorrhoeae strains FA1090 and MS11mkC

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    Marcia Metzgar Hobbs

    2011-05-01

    Full Text Available Experimental infection of male volunteers with Neisseria gonorrhoeae is safe and reproduces the clinical features of naturally acquired gonococcal urethritis. Human inoculation studies have helped define the natural history of experimental infection with two well-characterized strains of N. gonorrhoeae, FA1090 and MS11mkC. The human model has proved useful for testing the importance of putative gonococcal virulence factors for urethral infection in men. Studies with isogenic mutants have improved our understanding of the requirements for gonococcal LOS structures, pili, opacity proteins, IgA1 protease and the ability of infecting organisms to obtain iron from human transferrin and lactoferrin during uncomplicated urethritis. The model also presents opportunities to examine innate host immune responses that may be exploited or improved in development and testing of gonococcal vaccines. Here we review results to date with human experimental gonorrhea.

  2. Experimental rhinovirus infection in volunteers.

    Science.gov (United States)

    Bardin, P G; Sanderson, G; Robinson, B S; Holgate, S T; Tyrrell, D A

    1996-11-01

    Experimental viral disease studies in volunteers have clarified many aspects of the pathogenesis of human viral disease. Recently, interest has focused on rhinovirus-associated asthma exacerbations, and new volunteer studies have suggested that airway responsiveness (AR) is enhanced during a cold. For scientific, ethical and safety reasons, it is important to use validated methods for the preparation of a virus inoculum and that the particular virological characteristics and host responses should not be altered. We have prepared a new human rhinovirus (HRV) inoculum using recent guidelines and assessed whether disease characteristics (for example, severity of colds or changes in AR) were retained. Studies were conducted in 25 clinically healthy volunteers using a validated HRV inoculum in the first 17 and a new inoculum in the subsequent eight subjects. Severity of cold symptoms, nasal wash albumin levels and airway responsiveness were measured, and the new inoculum was prepared from nasal washes obtained during the cold. The new inoculum was tested using standard virological and serological techniques, as well as a polymerase chain reaction for Mycoplasma pneumoniae. No contaminating viruses or organisms were detected and the methods suggested were workable. Good clinical colds developed in 20 of the 25 subjects and median symptom scores were similar in the validated and new inoculum groups (18 and 17.5, respectively; p=0.19). All subjects shed virus, and there were no differences noted in viral culture scores, nasal wash albumin and rates of seroconversion in the two groups. Although airway responsiveness increased in both groups (p=0.02 and p=0.05), the degree of change was similar. We have performed experimental rhinovirus infection studies and demonstrated similar clinical disease in two inoculum groups. Amplified airway responsiveness was induced; continuing studies will define the mechanisms and suggest modes of treatment.

  3. Pathogenesis of reproductive failure induced by Trypanosoma vivax in experimentally infected pregnant ewes

    Science.gov (United States)

    2013-01-01

    The present study was aimed at investigating the effect of experimental infection by Trypanosoma vivax in different stages of pregnancy, determining the pathogenesis of reproductive failure, and confirming transplacental transmission. We used 12 pregnant ewes distributed into four experimental groups: G1, was formed by three ewes infected with T. vivax in the first third of pregnancy (30 days); G2 comprised three infected ewes in the final third of pregnancy (100 days); G3 and G4 were composed of three non-infected ewes with the same gestational period, respectively. Each ewe of G1 and G2 was inoculated with 1.25 × 105 tripomastigotes. Clinical examination, determination of parasitemia, serum biochemistry (albumin, total protein, glucose, cholesterol, and urea), packed cell volume (PCV), serum progesterone, and pathological examination were performed. Placenta, amniotic fluid, blood and tissues from the fetuses and stillbirths were submitted to PCR. Two ewes of G1 (Ewe 1 and 3) presented severe infection and died in the 34th and 35th days post-infection (dpi), respectively; but both fetuses were recovered during necropsy. In G2, Ewe 5 aborted two fetuses on the 130th day (30 dpi) of pregnancy; and Ewe 6 aborted one fetus in the 140th day (40 dpi) of gestation. Ewes 2 and 4 delivered two weak lambs that died five days after birth. Factors possibly involved with the reproductive failure included high parasitemia, fever, low PCV, body score, serum glucose, total protein, cholesterol, and progesterone. Hepatitis, pericarditis, and encephalitis were observed in the aborted fetuses. The presence of T. vivax DNA in the placenta, amniotic fluid, blood, and tissues from the fetuses confirms the transplacental transmission of the parasite. Histological lesion in the fetuses and placenta also suggest the involvement of the parasite in the etiopathogenesis of reproductive failure in ewes. PMID:23289625

  4. Endoscopic Endonasal Transsphenoidal Drainage of a Spontaneous Candida glabrata Pituitary Abscess.

    Science.gov (United States)

    Strickland, Ben A; Pham, Martin; Bakhsheshian, Joshua; Carmichael, John; Weiss, Martin; Zada, Gabriel

    2018-01-01

    Noniatrogenic pituitary abscess remains a rare clinical entity, and is the indication for surgery in abscess caused by Candida species, and also provide an intraoperative video showing the endoscopic management of this pathology. A 33-year-old woman presented with headache, hypopituitarism, and vision loss in the setting of diabetic ketoacidosis, and was found to have multiple abscesses in the liver, lung, kidney, and uterus. Brain magnetic resonance imaging revealed a 15-mm cystic sellar mass with restricted diffusion. The patient underwent urgent evacuation of the abscess via an endoscopic endonasal transsphenoidal route, with obvious purulent material filling the sella, later identified as Candida glabrata. Antimicrobial therapy was refined appropriately, and she exhibited significant improvement in neurologic function, although endocrinopathy has persisted. With timely management, including a combination of surgical drainage and appropriate antimicrobial therapy, neurologic outcomes are good in most cases of pituitary abscess; however, endocrinopathy often does not improve. Although most reported cases with identified causative organisms speciate bacteria, some cases are of fungal etiology and require different antimicrobial agents. This further underscores the importance of identifying the causative agent. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Antifungal susceptibility testing of Candida species isolated from the immunocompromised patients admitted to ten university hospitals in Iran: comparison of colonizing and infecting isolates.

    Science.gov (United States)

    Badiee, Parisa; Badali, Hamid; Boekhout, Teun; Diba, Kambiz; Moghadam, Abdolkarim Ghadimi; Hossaini Nasab, Ali; Jafarian, Hadis; Mohammadi, Rasoul; Mirhendi, Hossein; Najafzadeh, Mohammad Javad; Shamsizadeh, Ahmad; Soltani, Jafar

    2017-11-21

    Antifungal susceptibility testing is a subject of interest in the field of medical mycology. The aim of the present study were the distributions and antifungal susceptibility patterns of various Candida species isolated from colonized and infected immunocompromised patients admitted to ten university hospitals in Iran. In totally, 846 Candida species were isolated from more than 4000 clinical samples and identified by the API 20 C AUX system. Antifungal susceptibility testing was performed by broth microdilution method according to CLSI. The most frequent Candida species isolated from all patients was Candida albicans (510/846). The epidemiological cutoff value and percentage of wild-type species for amphotericin B and fluconazole in Candida albicans, Candida tropicalis, Candida glabrata and Candida krusei were 0.5 μg/ml (95%) and 4 μg/ml (96%); 1 μg/ml (95%) and 8 μg/ml (95%); 0.5 μg/ml (99%) and 19 μg/ml (98%); and 4 μg/ml (95%) and 64 μg/ml (95%), respectively. The MIC90 and epidemiological cutoff values to posaconazole in Candida krusei were 0.5 μg/ml. There were significant differences between infecting and colonizing isolates of Candida tropicalis in MIC 90 values of amphotericin B, and isolates of Candida glabrata in values of amphotericin B, caspofungin, and voriconazole (P Candida species (colonizing and infecting isolates) in immunocompromised patients are not the same and acquired resistance was seen in some species.

  6. Amphotericin B and caspofungin resistance in Candida glabrata isolates recovered from a critically ill patient

    DEFF Research Database (Denmark)

    Krogh-Madsen, Mikkel; Arendrup, Maiken Cavling; Heslet, Lars

    2006-01-01

    BACKGROUND: Consecutive Candida glabrata isolates recovered from a patient in an intensive care unit were resistant to amphotericin B (minimum inhibitory concentration, up to 32 mu g/mL; determined by Etest [AB Biodisk]). Analyses at the national reference laboratory showed that some isolates were...... also resistant to azoles and caspofungin. In this study, 4 isolates were studied thoroughly using susceptibility assays and a mouse model and to determine clonality. METHODS: Different broth microdilution tests, Etests, and time-kill studies for antifungals were performed in different media. Three...... isolates obtained from nonrelated patients, and a reference strain. RESULTS: The murine model indicated that 1 isolate was resistant to amphotericin B, 1 had intermediate susceptibility, and 1 was fully susceptible. Two of the 3 isolates were resistant to caspofungin. Microdilution methods did not reliably...

  7. Small sarcocysts can be a feature of experimental infections with Sarcocystis neurona merozoites.

    Science.gov (United States)

    Marsh, Antoinette E; Chaney, Sarah B; Howe, Daniel K; Saville, William J; Reed, Stephen M

    2017-10-15

    Several reports indicate the presence of small tissue cysts associated with Sarcocystis neurona infections. Several failed attempts to develop tissue cysts in potential intermediate host using in vitro derived parasites originally isolated from horses with equine protozoal myeloencephalitis suggest that the experimental methods to achieve bradyzoites with those isolates was not possible. Those prior studies reported the lack of detectable sarcocysts based on histology and in vivo feeding trials. A recent report of successful production and detection of small sarcocysts triggered us to review archived tissues from earlier experimental infection studies. The retrospective review sought to determine if small sized sarcocysts were not detected due to their relatively smaller size and infrequency as compared to larger sized sarcocysts produced with other isolates in these experimental inoculation trials. Tissues from two prior in vivo inoculation studies, involving in vitro-produced parasites inoculated into laboratory-reared cats and raccoons, were re-examined by immunohistochemistry staining to more easily detect the tissue cysts. In the experimental cat study no small tissue cysts were seen, consistent with the original publication results. However, in the experimental raccoon study, one raccoon inoculated with an EPM-derived isolate, SN-UCD1, had small sarcocysts not reported in the original publication. This retrospective study suggests that much closer scrutiny of tissues, including the use of immunohistochemistry on tissue sections is required to detect the smaller S. neurona sarcocysts associated with the experimental inoculations of the isolates originally derived from horses with EPM. Published by Elsevier B.V.

  8. Padronização da técnica de marcação de caramujos Biomphalaria glabrata com radioestroncio Standardization of the technic of labeling Biomphalaria glabrata snails with radiostrontium

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    Miguel Archanjo Muniz Leal

    1982-06-01

    Full Text Available Caramujos Biomphalaria glabrata vetor da esquistossomose mansônica, utilizados em experiências de campo, nas quais se faz necessário sua identificação e/ou localização, devem ser marcados com radioestrôncio 85,89 ou 90. Sua marcação em laboratório deve ser procedida em aquários de vidro sem fundo de areia, temperatura de 26,5 ± 1ºC, pH ajustado ao valor encontrado no "habitat", aeração artificial, alimentação diária com pequenas porções de alface fresca e 25 ml de água desionizada por caramujo. O nível de atividade no caramujo pode ser estimado sabendo-se que para o tempo de marcação de 5 dias e atividades iniciais de 0,036 a 0,362 µCi/ml, a captação global para grupos de 10 caramujos está em torno de 84 ± 2% e que a marcação em água contendo cálcio, praticamente não se altera até concentrações de 50ppm. A sobrevida ao longo de nove semanas não foi influenciada pela presença de radioestrôncio no caramujo.The vector snail of schistosomiasis mansoni, Biomphalaria glabrata, when used in field experiments in which it is to be recovered and/or located, should be labeled with radiostrontium 85,89 or 90. The labeling in laboratory should be carried out in an artificially aerated aquarium with 25 ml of deionized water per snail, without sand at the bottom, at a temperature of 26.5 ± 1ºC, pH adjusted to the value found in the natural habitat, and the snails should be fed with small pieces of fresh lettuce. The level of snail activity can be estimated taking into account that, for a labeling time of 5 days and initial activities of 0.036 to 0.362 µCi/ml, the global uptake for groups of 10 snails is about 84 ± 2% and that labeling in calcium-containing water parctically does not change till a 50 ppm concentration. Survival along 9 weeks was not influenced by the presence of radiostrontium in the snail.

  9. Transplacental and oral transmission of wild-type bluetongue virus serotype 8 in cattle after experimental infection

    NARCIS (Netherlands)

    Backx, A.; Heutink, C.G.; Rooij, van E.M.A.; Rijn, van P.A.

    2009-01-01

    Potential vertical transmission of wild-type bluetongue virus serotype 8 (BTV-8) in cattle was explored in this experiment. We demonstrated transplacental transmission of wild-type BTV-8 in one calf and oral infection with BTV-8 in another calf. Following the experimental BTV-8 infection of seven

  10. The Strategy to Survive Primary Malaria Infection: An Experimental Study on Behavioural Changes in Parasitized Birds.

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    Andrey Mukhin

    Full Text Available Avian malaria parasites (Haemosporida, Plasmodium are of cosmopolitan distribution, and they have a significant impact on vertebrate host fitness. Experimental studies show that high parasitemia often develops during primary malaria infections. However, field studies only occasionally reveal high parasitemia in free-living birds sampled using the traditional methods of mist-netting or trapping, and light chronic infections predominate. The reason for this discrepancy between field observation and experimental data remains insufficiently understood. Since mist-netting is a passive capture method, two main parameters determine its success in sampling infected birds in wildlife, i. e. the presence of parasitized birds at a study site and their mobility. In other words, the trapping probability depends on the survival rate of birds and their locomotor activity during infection. Here we test (1 the mortality rate of wild birds infected with Plasmodium relictum (the lineage pSGS1, (2 the changes in their behaviour during presence of an aerial predator, and (3 the changes in their locomotor activity at the stage of high primary parasitemia.We show that some behavioural features which might affect a bird's survival during a predator attack (time of reaction, speed of flush flight and take off angle did not change significantly during primary infection. However, the locomotor activity of infected birds was almost halved compared to control (non-infected birds during the peak of parasitemia. We report (1 the markedly reduced mobility and (2 the 20% mortality rate caused by P. relictum and conclude that these factors are responsible for the underrepresentation of birds in mist nets and traps during the stage of high primary parasitemia in wildlife. This study indicates that the widespread parasite, P. relictum (pSGS1 influences the behaviour of birds during primary parasitemia. Experimental studies combined with field observations are needed to better

  11. The Strategy to Survive Primary Malaria Infection: An Experimental Study on Behavioural Changes in Parasitized Birds

    Science.gov (United States)

    Mukhin, Andrey; Palinauskas, Vaidas; Platonova, Elena; Kobylkov, Dmitry; Vakoliuk, Irina; Valkiūnas, Gediminas

    2016-01-01

    Avian malaria parasites (Haemosporida, Plasmodium) are of cosmopolitan distribution, and they have a significant impact on vertebrate host fitness. Experimental studies show that high parasitemia often develops during primary malaria infections. However, field studies only occasionally reveal high parasitemia in free-living birds sampled using the traditional methods of mist-netting or trapping, and light chronic infections predominate. The reason for this discrepancy between field observation and experimental data remains insufficiently understood. Since mist-netting is a passive capture method, two main parameters determine its success in sampling infected birds in wildlife, i. e. the presence of parasitized birds at a study site and their mobility. In other words, the trapping probability depends on the survival rate of birds and their locomotor activity during infection. Here we test (1) the mortality rate of wild birds infected with Plasmodium relictum (the lineage pSGS1), (2) the changes in their behaviour during presence of an aerial predator, and (3) the changes in their locomotor activity at the stage of high primary parasitemia.We show that some behavioural features which might affect a bird's survival during a predator attack (time of reaction, speed of flush flight and take off angle) did not change significantly during primary infection. However, the locomotor activity of infected birds was almost halved compared to control (non-infected) birds during the peak of parasitemia. We report (1) the markedly reduced mobility and (2) the 20% mortality rate caused by P. relictum and conclude that these factors are responsible for the underrepresentation of birds in mist nets and traps during the stage of high primary parasitemia in wildlife. This study indicates that the widespread parasite, P. relictum (pSGS1) influences the behaviour of birds during primary parasitemia. Experimental studies combined with field observations are needed to better understand the

  12. Experimental infection and transmission of Leishmania by Lutzomyia cruzi (Diptera: Psychodidae: Aspects of the ecology of parasite-vector interactions.

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    Everton Falcão de Oliveira

    2017-02-01

    Full Text Available Several parameters should be addressed before incriminating a vector for Leishmania transmission. Those may include its ability to become infected by the same Leishmania species found in humans, the degree of attractiveness for reservoirs and humans and capacity to sustain parasite infection under laboratory conditions. This study evaluated the vectorial capacity of Lutzomyia cruzi for Leishmania infantum and gathered information on its ability to harbor L. amazonensis. Laboratory-reared Lu. cruzi were infected experimentally by feeding them on dogs infected naturally with L. infantum and hamsters infected with L. amazonensis. Sand fly attractiveness to dogs and humans was determined using wild caught insects. The expected daily survival of infected Lu. cruzi, the duration of the gonotrophic cycle, and the extrinsic incubation period were also investigated for both parasites. Vector competence was investigated for both Leishmania species. The mean proportion of female sand flies that fed on hosts was 0.40. For L. infantum and L. amazonensis, Lu. cruzi had experimental infection rates of 10.55% and 41.56%, respectively. The extrinsic incubation period was 3 days for both Leishmania species, regardless of the host. Survival expectancy of females infected with L. infantum and L. amazonensis after completing the gonotrophic cycle was 1.32 and 0.43, respectively. There was no association between L. infantum infection and sand fly longevity, but L. amazonensis-infected flies had significantly greater survival probabilities. Furthermore, egg-laying was significantly detrimental to survival. Lu. cruzi was found to be highly attracted to both dogs and humans. After a bloodmeal on experimentally infected hosts, both parasites were able to survive and develop late-stage infections in Lu. cruzi. However, transmission was demonstrated only for L. amazonensis-infected sand flies. In conclusion, Lu. cruzi fulfilled several of the requirements of vectorial

  13. Experimental infection and transmission of Leishmania by Lutzomyia cruzi (Diptera: Psychodidae): Aspects of the ecology of parasite-vector interactions.

    Science.gov (United States)

    Falcão de Oliveira, Everton; Oshiro, Elisa Teruya; Fernandes, Wagner de Souza; Murat, Paula Guerra; Medeiros, Márcio José de; Souza, Alda Izabel; Oliveira, Alessandra Gutierrez de; Galati, Eunice Aparecida Bianchi

    2017-02-01

    Several parameters should be addressed before incriminating a vector for Leishmania transmission. Those may include its ability to become infected by the same Leishmania species found in humans, the degree of attractiveness for reservoirs and humans and capacity to sustain parasite infection under laboratory conditions. This study evaluated the vectorial capacity of Lutzomyia cruzi for Leishmania infantum and gathered information on its ability to harbor L. amazonensis. Laboratory-reared Lu. cruzi were infected experimentally by feeding them on dogs infected naturally with L. infantum and hamsters infected with L. amazonensis. Sand fly attractiveness to dogs and humans was determined using wild caught insects. The expected daily survival of infected Lu. cruzi, the duration of the gonotrophic cycle, and the extrinsic incubation period were also investigated for both parasites. Vector competence was investigated for both Leishmania species. The mean proportion of female sand flies that fed on hosts was 0.40. For L. infantum and L. amazonensis, Lu. cruzi had experimental infection rates of 10.55% and 41.56%, respectively. The extrinsic incubation period was 3 days for both Leishmania species, regardless of the host. Survival expectancy of females infected with L. infantum and L. amazonensis after completing the gonotrophic cycle was 1.32 and 0.43, respectively. There was no association between L. infantum infection and sand fly longevity, but L. amazonensis-infected flies had significantly greater survival probabilities. Furthermore, egg-laying was significantly detrimental to survival. Lu. cruzi was found to be highly attracted to both dogs and humans. After a bloodmeal on experimentally infected hosts, both parasites were able to survive and develop late-stage infections in Lu. cruzi. However, transmission was demonstrated only for L. amazonensis-infected sand flies. In conclusion, Lu. cruzi fulfilled several of the requirements of vectorial capacity for L. infantum

  14. Highly pathogenic avian influenza virus (H5N1) in experimentally infected adult mute swans.

    Science.gov (United States)

    Kalthoff, Donata; Breithaupt, Angele; Teifke, Jens P; Globig, Anja; Harder, Timm; Mettenleiter, Thomas C; Beer, Martin

    2008-08-01

    Adult, healthy mute swans were experimentally infected with highly pathogenic avian influenza virus A/Cygnus cygnus/Germany/R65/2006 subtype H5N1. Immunologically naive birds died, whereas animals with preexisting, naturally acquired avian influenza virus-specific antibodies became infected asymptomatically and shed virus. Adult mute swans are highly susceptible, excrete virus, and can be clinically protected by preexposure immunity.

  15. Processing of predicted substrates of fungal Kex2 proteinases from Candida albicans, C. glabrata, Saccharomyces cerevisiae and Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Bader Oliver

    2008-07-01

    Full Text Available Abstract Background Kexin-like proteinases are a subfamily of the subtilisin-like serine proteinases with multiple regulatory functions in eukaryotes. In the yeast Saccharomyces cerevisiae the Kex2 protein is biochemically well investigated, however, with the exception of a few well known proteins such as the α-pheromone precursors, killer toxin precursors and aspartic proteinase propeptides, very few substrates are known. Fungal kex2 deletion mutants display pleiotropic phenotypes that are thought to result from the failure to proteolytically activate such substrates. Results In this study we have aimed at providing an improved assembly of Kex2 target proteins to explain the phenotypes observed in fungal kex2 deletion mutants by in vitro digestion of recombinant substrates from Candida albicans and C. glabrata. We identified CaEce1, CA0365, one member of the Pry protein family and CaOps4-homolog proteins as novel Kex2 substrates. Conclusion Statistical analysis of the cleavage sites revealed extended subsite recognition of negatively charged residues in the P1', P2' and P4' positions, which is also reflected in construction of the respective binding pockets in the ScKex2 enzyme. Additionally, we provide evidence for the existence of structural constrains in potential substrates prohibiting proteolysis. Furthermore, by using purified Kex2 proteinases from S. cerevisiae, P. pastoris, C. albicans and C. glabrata, we show that while the substrate specificity is generally conserved between organisms, the proteinases are still distinct from each other and are likely to have additional unique substrate recognition.

  16. Age-dependent differences in cytokine and antibody responses after experimental RSV infection in a bovine model

    DEFF Research Database (Denmark)

    Grell, S.N.; Riber, Ulla; Tjørnehøj, Kirsten

    2005-01-01

    Respiratory syncytial virus (RSV) causes severe respiratory disease in both infants and calves. As in humans, bovine RSV (BRSV) infections are most severe in the first 6 months of life. In this study, experimental infection with BRSV was performed in calves aged 1-5, 9-16 or 32-37 weeks. Compared...

  17. Alteration in the endogenous intestinal flora of swiss webster mice by experimental Angiostrongylus costaricensis infection

    Directory of Open Access Journals (Sweden)

    Vandack Nobre

    2004-11-01

    Full Text Available The association between worm infections and bacterial diseases has only recently been emphasized. This study examined the effect of experimental Angiostrongylus costaricensis infection on endogenous intestinal flora of Swiss Webster mice. Eight mice aging six weeks were selected for this experiment. Four were infected with A. costaricensis and the other four were used as controls. Twenty eight days after the worm infection, all mice in both groups were sacrificed and samples of the contents of the ileum and colon were obtained and cultured for aerobic and anaerobic bacteria. In the mice infected with A. costaricensis there was a significant increase in the number of bacteria of the endogenous intestinal flora, accompanied by a decrease in the number of Peptostreptococcus spp. This alteration in the intestinal flora of mice infected by the nematode may help to understand some bacterial infections described in humans.

  18. Experimental Infection of Taenia saginata eggs in Bali Cattle: Distribution and Density of Cysticercus bovis

    Directory of Open Access Journals (Sweden)

    Nyoman Sadra Dharmawan

    2009-12-01

    Full Text Available The objective of this study was to observe the development, distribution, and infection density ofTaenia saginata metacestodes in Bali cattle. Three Bali cattle were experimentally infected with T. saginataeggs which were collected from taeniasis patients. The experimental animal was inoculated with : i1000,00 T. saginata; ii 500,000 eggs; and iii 1,000,000 eggs, respectivelly 100,000 (cattle 1, 500,000(cattle 2, and 1,000,000 (cattle 3 T. saginata eggs, respectively. To observe the development of cysticerci,all cattle were slaughtered at 24 weeks post infection. To observe their distribution and density, slicingwas done to the cattle?s tissues. The study results showed that cysts were found distributed to all muscletissues and some visceral organs such as heart, diaphragm, lungs, and kidney of the cattle infected with100,000 and 500,000 T. saginata eggs. Density of the cyst was in the range of 11 to 95 cysts per 100 gramsof tissue. The highest density was noted in the heart (58/100 grams and in diaphragm (55/100 grams.This study has confirmed that T. saginata eggs derived from taeniasis patient in Bali, if infected to Balicattle can develop and spread to all muscle tissues and some visceral organs. From this study it wasconcluded that it is necessary to include the heart in the meat inspection at slaughter house for possibilityof T. saginata cyst infection.$?

  19. Effects of Co60 gamma radiation on Biomphalaria glabrata (Say, 1818) Embryo. II. Malformations

    International Nuclear Information System (INIS)

    Okazaki, K.; Kawano, T.

    1990-01-01

    The morphogenetic effects of ionizing radiation were investigated in Biomphalaria glabrata embryos irradiated in the cleavage, blastula, gastrula, young trochophore and trochophore stages with 5 to 25 Gy doses of 60 CO gamma radiation. The number of malformed embryos rapidly increased with increasing radiation dose, reaching a maximum between 5th to 8th day after irradiation in all stages analyzed. Susceptibility to malformation induction was higher the younger than the age of the irradiated embryo. However, for the cleavage stage the frequency of malformed embryos was inversely proportional to radiation dose for the same radiation dose. Several types of morphogenetic malformations were obtained, among then cephalic malformations, exogastrula, shell malformations and embryos with everted stomodeum, unspecific malformations being the most frequent. The results show that the types of malformation induced by radiation probably are not radiation-specific and do not depend on the dose applied [pt

  20. Intestinal colonization of broiler chickens by Campylobacter spp. in an experimental infection study

    DEFF Research Database (Denmark)

    Bahrndorff, Simon; Garcia Clavero, Ana Belén; Vigre, Håkan

    2015-01-01

    Consumption of poultry meat is considered as one of the main sources of human campylobacteriosis, and there is clearly a need for new surveillance and control measures based on quantitative data on Campylobacter spp. colonization dynamics in broiler chickens. We conducted four experimental...... infection trials, using four isolators during each infection trial to evaluate colonization of individual broiler chickens by Campylobacter jejuni over time. Individual and pooled faecal samples were obtained at days 4, 7 and 12 post-inoculation (p.i.) and caecal samples at day 12 p.i. There were large...

  1. The problems of urinary tract infections with Candida spp. aetiology in women.

    Science.gov (United States)

    Tomczak, Hanna; Szałek, Edyta; Grześkowiak, Edmund

    2014-08-29

    Urinary tract infections (UTIs) in women are a growing clinical concern. The most frequent risk factors of UTIs with fungal aetiology in women are: antibiotic therapy (especially broad-spectrum antibiotics), immunosuppressive therapy, diabetes, malnutrition, pregnancy, and frequent intercourse. The aim of the study was to analyse urinary tract infections with Candida spp. aetiology in women hospitalised at the Clinical Hospital in Poznań, Poland, between 2009 and 2011. The investigations revealed that as many as 71% of positive urine cultures with Candida fungi came from women. The following fungi were most frequently isolated from the patients under analysis: C. albicans (47%), C. glabrata (31%), C. tropicalis (6%), C. krusei (3%). In order to diagnose a UTI the diagnosis cannot be based on a single result of a urine culture. Due to the small number of antifungal drugs and high costs of treatment, antifungal drugs should be applied with due consideration and care.

  2. Phenotypic characterisation of cell populations in the brains of horses experimentally infected with West Nile virus.

    Science.gov (United States)

    Delcambre, G H; Liu, J; Streit, W J; Shaw, G P J; Vallario, K; Herrington, J; Wenzlow, N; Barr, K L; Long, M T

    2017-11-01

    West Nile virus (WNV), a mosquito borne member of the Flaviviridae, is one of the most commonly diagnosed agents of viral encephalitis in horses and people worldwide. A cassette of markers for formalin-fixed paraffin-embedded tissue and an archive of tissues from experimental infections in the horse were used to investigate the equine neuroimmune response to WNV meningoencephalomyelitis to phenotype the early response to WNV infection in the horse. Quantitative analysis using archived tissue from experimentally infected horses. The thalamus and hindbrain from 2 groups of 6 horses were compared and consisted of a culture positive tissues from WNV experimentally horses, in the other, normal horses. Formalin-fixed paraffin-embedded tissue from the thalamus and hindbrain were immunolabeled for microglia, astrocytes, B cells, macrophages/neutrophils, CD3 + T cells. Fresh frozen tissues were immunolabeled for CD4 + and CD8 + T lymphocyte cell markers. Cell counts were obtained using a computer software program. Differences, after meeting assumptions of abnormality, were computed using a general linear model with a Tukey test (Phorses, Iba-1 + microglia, CD3 + T lymphocyte and MAC387 + macrophage staining were significantly increased. The T cell response for the WNV-challenged horses was mixed, composed of CD4 + and CD8 + T lymphocytes. A limited astrocyte response was also observed in WNV-challenged horses, and MAC387 + and B cells were the least abundant cell populations. The results of this study were limited by a single collection time post-infection. Furthermore, a comprehensive analysis of cellular phenotypes is needed for naturally infected horses. Unfortunately, in clinical horses, there is high variability of sampling in terms of days post-infection and tissue handling. The data show that WNV-challenged horses recruit a mixed T cell population at the onset of neurologic disease. © 2017 EVJ Ltd.

  3. Experimental infection of Leishmania (L. chagasi in a cell line derived from Lutzomyia longipalpis (Diptera:Psychodidae

    Directory of Open Access Journals (Sweden)

    Felio J Bello

    2005-10-01

    Full Text Available The present work describes the in vitro infection of a cell line Lulo, derived from Lutzomyia longipalpis embryonic tissue, by Leishmania chagasi promastigotes. This infection process is compared with a parallel one developed using the J774 cell line. The L. chagasi MH/CO/84/CI-044B strain was used for experimental infection in two cell lines. The cells were seeded on glass coverslips in 24-well plates to reach a final number of 2 x 10(5 cells/well. Parasites were added to the adhered Lulo and J774 cells in a 10:1 ratio and were incubated at 28 and 37ºC respectively. After 2, 4, 6, 8, and 10 days post-infection, the cells were extensively washed with PBS, fixed with methanol, and stained with Giemsa. The number of internalized parasites was determined by counting at least 400 cultured cells on each coverslip. The results showed continuous interaction between L. chagasi promastigotes with the cell lines. Some ultrastructural characteristics of the amastigote forms were observed using transmission electron microscopy. The highest percentage of infection in Lulo cells was registered on day 6 post-infection (29.6% and on day 4 in the J774 cells (51%. This work shows similarities and differences in the L. chagasi experimental infection process in the two cell lines. However, Lulo cells emerge as a new model to study the life-cycle of this parasite.

  4. Respiratory disease in ball pythons (Python regius) experimentally infected with ball python nidovirus.

    Science.gov (United States)

    Hoon-Hanks, Laura L; Layton, Marylee L; Ossiboff, Robert J; Parker, John S L; Dubovi, Edward J; Stenglein, Mark D

    2018-04-01

    Circumstantial evidence has linked a new group of nidoviruses with respiratory disease in pythons, lizards, and cattle. We conducted experimental infections in ball pythons (Python regius) to test the hypothesis that ball python nidovirus (BPNV) infection results in respiratory disease. Three ball pythons were inoculated orally and intratracheally with cell culture isolated BPNV and two were sham inoculated. Antemortem choanal, oroesophageal, and cloacal swabs and postmortem tissues of infected snakes were positive for viral RNA, protein, and infectious virus by qRT-PCR, immunohistochemistry, western blot and virus isolation. Clinical signs included oral mucosal reddening, abundant mucus secretions, open-mouthed breathing, and anorexia. Histologic lesions included chronic-active mucinous rhinitis, stomatitis, tracheitis, esophagitis and proliferative interstitial pneumonia. Control snakes remained negative and free of clinical signs throughout the experiment. Our findings establish a causal relationship between nidovirus infection and respiratory disease in ball pythons and shed light on disease progression and transmission. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  5. Impact of the post-weaning parasitism history on an experimental Haemonchus contortus infection in Creole goat kids.

    Science.gov (United States)

    Ceï, W; Mahieu, M; Philibert, L; Arquet, R; Alexandre, G; Mandonnet, N; Bambou, J C

    2015-01-15

    Gastrointestinal nematode (GIN) infections have an important negative impact on small ruminant production. The selection of genotypes resistant to these parasitic infections is a promising alternative control strategy. Thus, resistance against GIN is an important component of small ruminant breeding schemes, based on phenotypic measurements of resistance in immune mature infected animals. In this study we evaluated both the impact of the post-weaning parasitism history on the response to an experimental Haemonchus contortus infection of resistant and susceptible Creole kids chosen on the basis of their estimated breeding value, and the interaction with the kid's genetic status. During the post-weaning period (from 3 months until 7 months of age) Creole kids were reared at pasture according to four different levels of a mixed rotational stocking system with Creole cattle: 100% (control), 75% (GG75), 50% (GG50), and 25% (GG25) of the total stocking rate of the pasture. The level of infection of the kids decreased significantly at 50% and 25% of the total stocking rate. After the post-weaning period at pasture, at 11 months of age kids were experimentally infected with H. contortus. The faecal egg counts (FEC) were significantly lower in the groups showing the highest FEC at pasture. This result suggests that a degree of protection against an experimental H. contortus infection occurred during the post-weaning period and was dependant on the level of parasitism. Interestingly, no interaction was observed between this level of protection and the genetic status. In conclusion, the level of post-weaning natural parasitism history at pasture would not influence the genetic status evaluation. More generally our results suggest that it would be better to expose kids to a high level of gastrointestinal parasitism during the post-weaning period in order to increase the basal level of resistance thereafter. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Overcoming the heterologous bias: An in vivo functional analysis of multidrug efflux transporter, CgCdr1p in matched pair clinical isolates of Candida glabrata

    International Nuclear Information System (INIS)

    Puri, Nidhi; Manoharlal, Raman; Sharma, Monika; Sanglard, Dominique; Prasad, Rajendra

    2011-01-01

    Research highlights: → First report to demonstrate an in vivo expression system of an ABC multidrug transporter CgCdr1p of C. glabrata. → First report on the structure and functional characterization of CgCdr1p. → Functional conservation of divergent but typical residues of CgCdr1p. → CgCdr1p elicits promiscuity towards substrates and has a large drug binding pocket with overlapping specificities. -- Abstract: We have taken advantage of the natural milieu of matched pair of azole sensitive (AS) and azole resistant (AR) clinical isolates of Candida glabrata for expressing its major ABC multidrug transporter, CgCdr1p for structure and functional analysis. This was accomplished by tagging a green fluorescent protein (GFP) downstream of ORF of CgCDR1 and integrating the resultant fusion protein at its native chromosomal locus in AS and AR backgrounds. The characterization confirmed that in comparison to AS isolate, CgCdr1p-GFP was over-expressed in AR isolates due to its hyperactive native promoter and the GFP tag did not affect its functionality in either construct. We observed that in addition to Rhodamine 6 G (R6G) and Fluconazole (FLC), a recently identified fluorescent substrate of multidrug transporters Nile Red (NR) could also be expelled by CgCdr1p. Competition assays with these substrates revealed the presence of overlapping multiple drug binding sites in CgCdr1p. Point mutations employing site directed mutagenesis confirmed that the role played by unique amino acid residues critical to ATP catalysis and localization of ABC drug transporter proteins are well conserved in C. glabrata as in other yeasts. This study demonstrates a first in vivo novel system where over-expression of GFP tagged MDR transporter protein can be driven by its own hyperactive promoter of AR isolates. Taken together, this in vivo system can be exploited for the structure and functional analysis of CgCdr1p and similar proteins wherein the arte-factual concerns encountered in using

  7. Overcoming the heterologous bias: An in vivo functional analysis of multidrug efflux transporter, CgCdr1p in matched pair clinical isolates of Candida glabrata

    Energy Technology Data Exchange (ETDEWEB)

    Puri, Nidhi; Manoharlal, Raman; Sharma, Monika [Membrane Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi 110 067 (India); Sanglard, Dominique [Institut de Microbiologie, Centre Hospitalier Universitaire Vaudois, 1011 Lausanne (Switzerland); Prasad, Rajendra, E-mail: rp47jnu@gmail.com [Membrane Biology Laboratory, School of Life Sciences, Jawaharlal Nehru University, New Delhi 110 067 (India)

    2011-01-07

    Research highlights: {yields} First report to demonstrate an in vivo expression system of an ABC multidrug transporter CgCdr1p of C. glabrata. {yields} First report on the structure and functional characterization of CgCdr1p. {yields} Functional conservation of divergent but typical residues of CgCdr1p. {yields} CgCdr1p elicits promiscuity towards substrates and has a large drug binding pocket with overlapping specificities. -- Abstract: We have taken advantage of the natural milieu of matched pair of azole sensitive (AS) and azole resistant (AR) clinical isolates of Candida glabrata for expressing its major ABC multidrug transporter, CgCdr1p for structure and functional analysis. This was accomplished by tagging a green fluorescent protein (GFP) downstream of ORF of CgCDR1 and integrating the resultant fusion protein at its native chromosomal locus in AS and AR backgrounds. The characterization confirmed that in comparison to AS isolate, CgCdr1p-GFP was over-expressed in AR isolates due to its hyperactive native promoter and the GFP tag did not affect its functionality in either construct. We observed that in addition to Rhodamine 6 G (R6G) and Fluconazole (FLC), a recently identified fluorescent substrate of multidrug transporters Nile Red (NR) could also be expelled by CgCdr1p. Competition assays with these substrates revealed the presence of overlapping multiple drug binding sites in CgCdr1p. Point mutations employing site directed mutagenesis confirmed that the role played by unique amino acid residues critical to ATP catalysis and localization of ABC drug transporter proteins are well conserved in C. glabrata as in other yeasts. This study demonstrates a first in vivo novel system where over-expression of GFP tagged MDR transporter protein can be driven by its own hyperactive promoter of AR isolates. Taken together, this in vivo system can be exploited for the structure and functional analysis of CgCdr1p and similar proteins wherein the arte-factual concerns

  8. Isolation of Trichophyton mentogrophytes var mentogrophytes from naturally infected laboratory albino rats: experimental infection and treatment in rabbits

    Directory of Open Access Journals (Sweden)

    N. A. Issa

    2009-01-01

    Full Text Available The present study demonstrated for the first time the occurrence of dermatophytosis in naturally infected rats and from asymptomatic and from breeding boxes of white rats kept in animal housing of college of Veterinary Medicine, University of Dohuk, Iraq. The prevalence rate of infection was (28%, clinically infected rats characterized by appearance of scaly ovoid type lesions with crusty edge and patch of hair loss mostly seen on the back, neck and face of the infected rats, itching was reported in some rats. Only one species of the trichophyton, T. mentogrophytes var mentogrophytes was isolated with growth rate (85.71% of samples collected from clinically infected rats, and (28.57% from asymptomatic and from breeding cages, the growth was observed within the 21 days at 25ºC on Sabouraud's Dextrose Agar. Lacto phenol cotton blue staining slides of T. mentogrophytes var mentogrophytes revealed both microconidia and macroconidia. Microconidia found in numerous numbers often in dense cluster which were hyaline, smooth walled and predominantly spherical to sub spherical in shape, varying numbers of chlamydoconidia. Spiral hyphae and smooth, thin walled clavate shaped multicelled macroconidia were also present. The study also dealt with experimental infection in rabbits with T. mentogrophytes var mentogrophytes and treated by two drugs, natural herbal preparation of acidic pomegranate (Punica granatum fruit and synthetic nystatine ointment. The complete recovery of lesions was recorded after 14 days and 21 days of topical application of a pomegranate and nystatine ointment for 5 successive days respectively.

  9. Experimental infection of highly pathogenic avian influenza virus H5N1 in black-headed gulls (Chroicocephalus ridibundus)

    OpenAIRE

    Ramis , Antonio; van Amerongen , Geert; van de Bildt , Marco; Leijten , Loneke; Vanderstichel , Raphael; Osterhaus , Albert; Kuiken , Thijs

    2014-01-01

    Historically, highly pathogenic avian influenza viruses (HPAIV) rarely resulted in infection or clinical disease in wild birds. However, since 2002, disease and mortality from natural HPAIV H5N1 infection have been observed in wild birds including gulls. We performed an experimental HPAIV H5N1 infection of black-headed gulls (Chroicocephalus ridibundus) to determine their susceptibility to infection and disease from this virus, pattern of viral shedding, clinical signs, pathological changes a...

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

    Directory of Open Access Journals (Sweden)

    Alyssa N Malone

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

  11. Avaliação do tipo de substrato e do período de armazenamento para a germinação de sementes de Cordia glabrata (Mart. DC. = Evaluation of substrate type and storage period for the germination of Cordia glabrata (Mart. DC seeds

    Directory of Open Access Journals (Sweden)

    Juliana Gadum

    2009-07-01

    Full Text Available Informações sobre a viabilidade e substratos adequados para a germinação de sementes de espécies arbóreas nativas são escassas. Cordia glabrata (louro-preto, espécie pioneira e de ampla distribuição, está incluída entre essas espécies. Suas característicaspermitem a utilização em ornamentação, na prática apícola e na medicina popular. Levando em conta seu potencial de utilização, este trabalho teve por objetivo avaliar as taxas de germinação, em diferentes substratos, de sementes recém-colhidas e armazenadas até 120 dias em laboratório, as quais foram coletadas no Pantanal do Rio Negro, Estado do Mato Grosso do Sul. Foram realizados testes com sementes recém-colhidas e após 30, 60, 90 e 120 dias de armazenamento, em três tipos de substrato (papel-filtro, solo arenoso e solo arenoso + matéria orgânica. Os resultados revelaram que C. glabrata possui pequena porcentagem de água (1,55% nas sementes, que decai com o período de armazenamento. Seu período de viabilidade é curto (90 dias e os melhores resultados foram obtidos com sementes recém-colhidas germinadas em papel-filtro (34%, com índice de velocidade de germinação de 10.Information about the feasibility and adequate substrates for the germination of seeds from native arboreous species are scarce. Cordiaglabrata (louro preto, a pioneering species with wide distribution, is included among these species. Its characteristics allow its use in ornamentation, apiculture and popular medicine. Considering its potential of utilization, this work had as objective to evaluate thegermination rates, in different substrates, of recently reaped seeds stored for up to 120 days in laboratory, which were collected in Pantanal of the Rio Negro, Mato Grosso do Sul State. Tests were done with recently reaped seeds and after 30, 60, 90 and 120 days ofstorage, in three kinds of substrate (paper filter, sandy soil and sandy soil + organic matter. The results revealed that C

  12. Synergistic mutual potentiation of antifungal activity of Zuccagnia punctata Cav. and Larrea nitida Cav. extracts in clinical isolates of Candida albicans and Candida glabrata.

    Science.gov (United States)

    Butassi, Estefanía; Svetaz, Laura A; Ivancovich, Juan J; Feresin, Gabriela E; Tapia, Alejandro; Zacchino, Susana A

    2015-06-01

    Zuccagnia punctata Cav. (Fabaceae) and Larrea nitida Cav. (Zygophyllaceae) are indistinctly or jointly used in traditional medicine for the treatment of fungal-related infections. Although their dichloromethane (DCM) extract have demonstrated moderate antifungal activities when tested on their own, antifungal properties of combinations of both plants have not been assessed previously. The aim of this study was to establish with statistical rigor whether Z. punctata (ZpE) and L. nitida DCM extract (LnE) interact synergistically against the clinically important fungi Candida albicans and Candida glabrata and to characterize the most synergistic combinations. For synergism assessment, the statistical-based Boik's design was applied. Eight ZpE-LnE fixed-ratio mixtures were prepared from four different months of 1 year and tested against Candida strains. Lϕ (Loewe index) of each mixture at different fractions affected (ϕ) allowed for the finding of the most synergistic combinations, which were characterized by HPLC fingerprint and by the quantitation of the selected marker compounds. Lϕ and confidence intervals were determined in vitro with the MixLow method, once the estimated parameters from the dose-response curves of independent extracts and mixtures, were obtained. Markers (four flavonoids for ZpE and three lignans for LnE) were quantified in each extract and their combinations, with a valid HPLC-UV method. The 3D-HPLC profiles of the most synergistic mixtures were obtained by HPLC-DAD. Three over four IC50ZpE/IC50LnE fixed-ratio mixtures displayed synergistic interactions at effect levels ϕ > 0.5 against C. albicans. The dosis of the most synergistic (Lϕ = 0.62) mixture was 65.96 µg/ml (ZpE = 28%; LnE = 72%) containing 8 and 36% of flavonoids and lignans respectively. On the other hand, one over four IC50ZpE/IC50LnE mixtures displays synergistic interactions at ϕ > 0.5 against C. glabrata. The dosis of the most synergistic (Lϕ = 0.67) mixture was 168

  13. Clinical presentation, convalescence, and relapse of rocky mountain spotted fever in dogs experimentally infected via tick bite.

    Directory of Open Access Journals (Sweden)

    Michael L Levin

    Full Text Available Rocky Mountain spotted fever (RMSF is a tick-borne disease caused by R. rickettsii in North and South America. Domestic dogs are susceptible to infection and canine RMSF can be fatal without appropriate treatment. Although clinical signs of R. rickettsii infection in dogs have been described, published reports usually include descriptions of either advanced clinical cases or experimental infections caused by needle-inoculation of cultured pathogen rather than by tick bite. The natural progression of a tick-borne R. rickettsii infection has not been studied in sufficient detail. Here, we provide a detailed description of clinical, hematological, molecular, and serological dynamics of RMSF in domestic dogs from the day of experimental exposure to infected ticks through recovery. Presented data indicate that neither the height/duration of fever nor detection of rickettsial DNA in dogs' blood by PCR are good indicators for clinical prognosis. Only the apex and subsequent subsidence of neutrophilia seem to mark the beginning of recovery and allow predicting a favorable outcome in Rickettsia-infected dogs, even despite the continuing persistence of mucosal petechiae and skin rash. On the other hand the appropriate (doxycycline antibiotic therapy of sufficient duration is crucial in prevention of RMSF relapses in dogs.

  14. Clinical presentation, convalescence, and relapse of rocky mountain spotted fever in dogs experimentally infected via tick bite.

    Science.gov (United States)

    Levin, Michael L; Killmaster, Lindsay F; Zemtsova, Galina E; Ritter, Jana M; Langham, Gregory

    2014-01-01

    Rocky Mountain spotted fever (RMSF) is a tick-borne disease caused by R. rickettsii in North and South America. Domestic dogs are susceptible to infection and canine RMSF can be fatal without appropriate treatment. Although clinical signs of R. rickettsii infection in dogs have been described, published reports usually include descriptions of either advanced clinical cases or experimental infections caused by needle-inoculation of cultured pathogen rather than by tick bite. The natural progression of a tick-borne R. rickettsii infection has not been studied in sufficient detail. Here, we provide a detailed description of clinical, hematological, molecular, and serological dynamics of RMSF in domestic dogs from the day of experimental exposure to infected ticks through recovery. Presented data indicate that neither the height/duration of fever nor detection of rickettsial DNA in dogs' blood by PCR are good indicators for clinical prognosis. Only the apex and subsequent subsidence of neutrophilia seem to mark the beginning of recovery and allow predicting a favorable outcome in Rickettsia-infected dogs, even despite the continuing persistence of mucosal petechiae and skin rash. On the other hand the appropriate (doxycycline) antibiotic therapy of sufficient duration is crucial in prevention of RMSF relapses in dogs.

  15. Inactivation of Candida glabrata by a humid DC argon discharge afterglow: dominant contributions of short-lived aqueous active species

    Science.gov (United States)

    Xiong, Qing; Liu, Hongbin; Lu, Weiping; Chen, Qiang; Xu, Le; Wang, Xia; Zhu, Qunlin; Zeng, Xue; Yi, Ping

    2017-05-01

    Plasma medicine applications are currently attracting significant interest all over the world. Bactericidal treatments of Candida glabrata cultured in saline suspension are performed in this study by a room-temperature reactive afterglow of a DC-driven argon discharge. Water vapor was added to the discharge to study the inactivation contributions of reactive hydrolytic species including OH and H2O2 transporting along the gas flow to the treated solutions. The inactivation results indicate that the dominant roles in the bactericidal treatments are played by the short-lived aqueous active species, but not the stable species like H2O2aq (aq indicates an aqueous species). Further analysis shows that the ·OHaq radicals play an important role in the inactivation process. The ·OHaq radicals in the suspension are mostly produced from the direct dissolution of the OH species in the reactive afterglow. With the increase of added water vapor content, the ·OHaq production increases and enhances the inactivation efficiency of C. glabrata. Furthermore, it is found that the ambient air diffusion shows essential effects on the bactericidal activity of the remote humid argon discharge. Higher bactericidal effects can be obtained in open-space treatments compared to in a controlled Ar + H2O gas atmosphere. Key active air-byproduct species are believed to be generated in the suspension during the treatments and contributing to the inactivation process. Based on chemical analysis, the peroxynitrous acid ONOOHaq is considered as the key antimicrobial air-byproduct species. These results indicate the important dependence of plasma biomedical effects on the processing environment, which finally relates to the critical contributions of the key reactive species formed therein.

  16. Facultative Sterol Uptake in an Ergosterol-Deficient Clinical Isolate of Candida glabrata Harboring a Missense Mutation in ERG11 and Exhibiting Cross-Resistance to Azoles and Amphotericin B

    Science.gov (United States)

    Hull, Claire M.; Parker, Josie E.; Bader, Oliver; Weig, Michael; Gross, Uwe; Warrilow, Andrew G. S.; Kelly, Diane E.

    2012-01-01

    We identified a clinical isolate of Candida glabrata (CG156) exhibiting flocculent growth and cross-resistance to fluconazole (FLC), voriconazole (VRC), and amphotericin B (AMB), with MICs of >256, >256, and 32 μg ml−1, respectively. Sterol analysis using gas chromatography-mass spectrometry (GC-MS) revealed that CG156 was a sterol 14α-demethylase (Erg11p) mutant, wherein 14α-methylated intermediates (lanosterol was >80% of the total) were the only detectable sterols. ERG11 sequencing indicated that CG156 harbored a single-amino-acid substitution (G315D) which nullified the function of native Erg11p. In heterologous expression studies using a doxycycline-regulatable Saccharomyces cerevisiae erg11 strain, wild-type C. glabrata Erg11p fully complemented the function of S. cerevisiae sterol 14α-demethylase, restoring growth and ergosterol synthesis in recombinant yeast; mutated CG156 Erg11p did not. CG156 was culturable using sterol-free, glucose-containing yeast minimal medium (glcYM). However, when grown on sterol-supplemented glcYM (with ergosta 7,22-dienol, ergosterol, cholestanol, cholesterol, Δ7-cholestenol, or desmosterol), CG156 cultures exhibited shorter lag phases, reached higher cell densities, and showed alterations in cellular sterol composition. Unlike comparator isolates (harboring wild-type ERG11) that became less sensitive to FLC and VRC when cultured on sterol-supplemented glcYM, facultative sterol uptake by CG156 did not affect its azole-resistant phenotype. Conversely, CG156 grown using glcYM with ergosterol (or with ergosta 7,22-dienol) showed increased sensitivity to AMB; CG156 grown using glcYM with cholesterol (or with cholestanol) became more resistant (MICs of 2 and >64 μg AMB ml−1, respectively). Our results provide insights into the consequences of sterol uptake and metabolism on growth and antifungal resistance in C. glabrata. PMID:22615281

  17. Fungal Urinary Tract Infection in Burn Patients‎

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    Suad Yousuf Aldorkee

    2017-11-01

    Full Text Available Background: Urinary tract infection is the most common hospital-acquired infection. Fungal species are unusual causes of urinary tract infection in healthy individuals, but common in the hospital setting or among patients with predisposing diseases and structural abnormalities of the kidney and collecting system. Burn patients are susceptible to nosocomial infections owing to the immunocompromising effects of burn injury, cutaneous and respiratory tract injury, prolonged intensive care unit stays and broad-spectrum antibiotic therapy. Objective: The study population includes adult patients of both genders who presented with different percentages of body burns. Urine sample was collected from each patient at the time of admission and weekly thereafter for 6 weeks and sent for general urine examination and urine culture to test for the possibility of fungal growth. Those who found to develop fungal UTI by urine culture during their hospitalization and had no infection at the time of admission were selected as subjects for our study. Results: 28 (18.6% patients had positive fungal culture during their hospitalization, 11 of them were males and 17 were females, the most common age of presentation was 41-50 years and the mean age ± SD was (44.4 ± 10.7 years. The most common isolated fungi were Candida albicans (64.3%, followed by Candida glabrata (21.4% and Candida tropicalis (7.1%. The majority of patients developed infection within the 2nd and 3rd weeks of hospitalization, however, those who presented with total body surface area burned > 40% developed an earlier infection within the 1st week. Female gender, urethral catheterization and diabetes mellitus were significantly associated with higher risk of infection as the P values were 0.03, 0.005 and 0.004 respectively. Conclusion: Fungal urinary tract infection occurred in 18.6% of burn patients. The most common causative fungi are candida species. Advanced age, female gender, high percentage of

  18. Antifungal susceptibility testing of Candida species isolated from the immunocompromised patients admitted to ten university hospitals in Iran: comparison of colonizing and infecting isolates

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    Parisa Badiee

    2017-11-01

    Full Text Available Abstract Background Antifungal susceptibility testing is a subject of interest in the field of medical mycology. The aim of the present study were the distributions and antifungal susceptibility patterns of various Candida species isolated from colonized and infected immunocompromised patients admitted to ten university hospitals in Iran. Methods In totally, 846 Candida species were isolated from more than 4000 clinical samples and identified by the API 20 C AUX system. Antifungal susceptibility testing was performed by broth microdilution method according to CLSI. Results The most frequent Candida species isolated from all patients was Candida albicans (510/846. The epidemiological cutoff value and percentage of wild-type species for amphotericin B and fluconazole in Candida albicans, Candida tropicalis, Candida glabrata and Candida krusei were 0.5 μg/ml (95% and 4 μg/ml (96%; 1 μg/ml (95% and 8 μg/ml (95%; 0.5 μg/ml (99% and 19 μg/ml (98%; and 4 μg/ml (95% and 64 μg/ml (95%, respectively. The MIC90 and epidemiological cutoff values to posaconazole in Candida krusei were 0.5 μg/ml. There were significant differences between infecting and colonizing isolates of Candida tropicalis in MIC 90 values of amphotericin B, and isolates of Candida glabrata in values of amphotericin B, caspofungin, and voriconazole (P < 0.05. Conclusions Our findings suggest that the susceptibility patterns of Candida species (colonizing and infecting isolates in immunocompromised patients are not the same and acquired resistance was seen in some species.

  19. Lab-scale preparations of Candida albicans and dual Candida albicans-Candida glabrata biofilms on the surface of medical-grade polyvinyl chloride (PVC) perfusion tube using a modified gravity-supported free-flow biofilm incubator (GS-FFBI).

    Science.gov (United States)

    Shao, Jing; Lu, KeQiao; Tian, Ge; Cui, YanYan; Yan, YuanYuan; Wang, TianMing; Zhang, XinLong; Wang, ChangZhong

    2015-02-01

    The assembly of a man-made gravity-supported free-flow biofilm incubator (GS-FFBI) was described, which was composed of a gas cushion injector and four incubators. The GS-FFBI had the characteristics of (i) a bottom-up flow direction, and (ii) lab-scale biofilm preparation without the use of a multichannel pump. Two opportunistic fungal strains, namely Candida albicans and Candida glabrata, were employed to incubate C. albicans and dual C. albicans-C. glabrata biofilms on the surface of medical-grade polyvinyl chloride perfusion tube. In terms of the results from {2, 3-bis (2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide} (XTT) assay, dry weight measurement, colony-forming unit counting, susceptibility test, and scanning electron microscopy, it was demonstrated that GS-FFBI could form both stable single and dual Candida biofilms with no significant variations among the four incubators or the three daily incubations within 21h, and could operate for at least 96h smoothly with no contamination of stock medium. The results also indicated, for the first time, that C. albicans and C. glabrata might be co-existent competitively and symbiotically in the dual biofilms with flowing media. GS-FFBI would be a useful device to study in vitro morphological and physiological features of microbial biofilms in the medical settings. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Experimental Infections of Oryzomys couesi with Sympatric Arboviruses from Mexico

    Science.gov (United States)

    Deardorff, Eleanor R.; Forrester, Naomi L.; Travassos da Rosa, Amelia P.; Estrada-Franco, Jose G.; Navarro-Lopez, Roberto; Tesh, Robert B.; Weaver, Scott C.

    2010-01-01

    Coues rice rat (Oryzomys couesi), a species abundant throughout Central America, was evaluated experimentally for the ability to serve as an amplifying host for three arboviruses: Patois (Bunyaviridae, Orthobunyavirus), Nepuyo (Orthobunyavirus), and Venezuelan equine encephalitis virus subtype ID (Togaviridae, Alphavirus). These three viruses have similar ecologies and are known to co-circulate in nature. Animals from all three cohorts survived infection and developed viremia with no apparent signs of illness and long-lasting antibodies. Thus, O. couesi may play a role in the general maintenance of these viruses in nature. PMID:20134016

  1. Redistribution of carbon flux in Torulopsis glabrata by altering vitamin and calcium level.

    Science.gov (United States)

    Liu, Liming; Li, Yin; Zhu, Yang; Du, Guocheng; Chen, Jian

    2007-01-01

    Manipulation of cofactor (thiamine, biotin and Ca(2+)) levels as a potential tool to redistribute carbon flux was studied in Torulopsis glabrata. With sub-optimization of vitamin in fermentation medium, the carbon flux was blocked at the key node of pyruvate, and 69 g/L pyruvate was accumulated. Increasing the concentrations of thiamine and biotin could selectively open the valve of carbon flux from pyruvate to pyruvate dehydrogenase complex, the pyruvate carboxylase (PC) pathway and the channel into the TCA cycle, leading to the over-production of alpha-ketoglutarate. In addition, the activity of PC was enhanced with Ca(2+) present in fermentation medium. By combining high concentration's vitamins and CaCO(3) as the pH buffer, a batch culture was conducted in a 7-L fermentor, with the pyruvate concentration decreased to 21.8 g/L while alpha-ketoglutarate concentration increased to 43.7 g/L. Our study indicated that the metabolic flux could be redistributed to overproduce desired metabolites with manipulating the cofactor levels. Furthermore, the manipulation of vitamin level provided an alternative tool to realize metabolic engineering goals.

  2. Pathogenesis of Candida albicans infections in the alternative chorio-allantoic membrane chicken embryo model resembles systemic murine infections.

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    Ilse D Jacobsen

    Full Text Available Alternative models of microbial infections are increasingly used to screen virulence determinants of pathogens. In this study, we investigated the pathogenesis of Candida albicans and C. glabrata infections in chicken embryos infected via the chorio-allantoic membrane (CAM and analyzed the virulence of deletion mutants. The developing immune system of the host significantly influenced susceptibility: With increasing age, embryos became more resistant and mounted a more balanced immune response, characterized by lower induction of proinflammatory cytokines and increased transcription of regulatory cytokines, suggesting that immunopathology contributes to pathogenesis. While many aspects of the chicken embryo response resembled murine infections, we also observed significant differences: In contrast to systemic infections in mice, IL-10 had a beneficial effect in chicken embryos. IL-22 and IL-17A were only upregulated after the peak mortality in the chicken embryo model occurred; thus, the role of the Th17 response in this model remains unclear. Abscess formation occurs frequently in murine models, whereas the avian response was dominated by granuloma formation. Pathogenicity of the majority of 15 tested C. albicans deletion strains was comparable to the virulence in mouse models and reduced virulence was associated with significantly lower transcription of proinflammatory cytokines. However, fungal burden did not correlate with virulence and for few mutants like bcr1Δ and tec1Δ different outcomes in survival compared to murine infections were observed. C. albicans strains locked in the yeast stage disseminated significantly more often from the CAM into the embryo, supporting the hypothesis that the yeast morphology is responsible for dissemination in systemic infections. These data suggest that the pathogenesis of C. albicans infections in the chicken embryo model resembles systemic murine infections but also differs in some aspects. Despite

  3. Immunodiagnosis of systemic aspergillosis. I. Antigenemia detected by radioimmunoassay in experimental infection

    International Nuclear Information System (INIS)

    Weiner, M.H.; Coats-Stephen, M.

    1979-01-01

    Because systemic aspergillosis is difficult to diagnose ante mortem, a study to improve immunodiagnosis was undertaken in a rabbit model of disseminated infection. We found that the predominant humoral response of infected animals was directed against four Aspergillus antigens identified by crossed immunoelectrophoresis. One of these antigens, a cell-wall carbohydrate, was purified by gel-filtration chromatography and was used to develop a radiommunoassay. The sensitivity of this assay was increased by testing for serum-bound antigen as well as for free antigen. When the sensitivity of the RIA was evaluated in the animal model, antigenemia was detected in 78% of 51 rabbits with disseminated infection and ante mortem in 86% of 42 rabbits with lethal infection. By contrast, with immunoprecipitin analysis only eight of 51 rabbits were positive for antigen, and six of 51 rabbits were positive for Aspergillus antibody. The specificity of the RIA was also tested. Negative controls for antigen included sera from 76 normal rabbits and sera from 25 rabbits with systemic candidiasis. The Candida control group is pertinent because 48% of these rabbits had specific Candida antigenemia detected by a mannan RIA. This study demonstrates that Aspergillus antigenemia occurs during the course of experimental disseminated aspergillosis and illustrates the potential of an Aspergillus antigen RIA for sensitive, specific immunodiagnosis of human infections

  4. Comportamento reológico de méis de florada de silvestre Rheological behavior of honey from Serjania glabrata flowers

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    Alexandre J. de M. Queiroz

    2007-04-01

    Full Text Available Estudou-se o comportamento reológico de méis de Apis mellifera produzidos no estado do Piauí, no semi-árido brasileiro, elaborados com florada predominante de silvestre (Serjania glabrata. As amostras foram coletadas ao longo do ano 2000, a partir de coletas realizadas por apicultores, associações e cooperativas de apicultura da região abrangida pelos municípios de Picos, Itainópolis, Vera Mendes e Isaías Coelho. Após as coletas as amostras passaram pelos processos de centrifugação, filtragem e decantação. As medidas reológicas foram feitas nas temperaturas de 20 a 40 ºC utilizando-se um viscosímetro Brookfield, modelo RVT. As leituras de velocidade de rotação e torque foram transformadas em valores de taxa de deformação e tensão de cisalhamento. Os dados de taxa de deformação e tensão de cisalhamento foram ajustados pelas equações da Lei-da-Potência e de Herschel-Bulkley. As amostras apresentaram comportamento pseudoplástico. As equações se ajustaram adequadamente aos resultados experimentais de tensão de cisalhamento em função da taxa de deformação. O aumento de temperatura reduziu a viscosidade aparente em níveis próximos de 80%. Os valores de viscosidade aparente foram bem ajustados por uma equação do tipo Arrhenius.The rheological behavior of Apis mellifera honey produced in the State of Piauí, in the Brazilian semi-arid, obtained with predominance of Serjania glabrata flowers was studied. The samples were collected in the year 2000 by beekeepers and associations and cooperatives of beekeepers in the area covered by the municipal districts of Picos, Itainópolis, Vera Mendes and Isaías Coelho. The samples were centrifuged, filtered, decanted and the rheological measures at temperatures of 20 to 40 ºC using a Brookfield Viscometer RVT model were made. The readings of rotation speed and torque were transformed in shear rate and shear stress values. The data of shear rate and shear stress were fitted

  5. Avaliação do tipo de substrato e do período de armazenamento para a germinação de sementes de Cordia glabrata (Mart. DC. - DOI: 10.4025/actascibiolsci.v31i3.1013 Evaluation of substrate type and storage period for the germination of Cordia glabrata (Mart. DC seeds - DOI: 10.4025/actascibiolsci.v31i3.1013

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    Juliana Gadum

    2009-07-01

    Full Text Available Informações sobre a viabilidade e substratos adequados para a germinação de sementes de espécies arbóreas nativas são escassas. Cordia glabrata (louro-preto, espécie pioneira e de ampla distribuição, está incluída entre essas espécies. Suas características permitem a utilização em ornamentação, na prática apícola e na medicina popular. Levando em conta seu potencial de utilização, este trabalho teve por objetivo avaliar as taxas de germinação, em diferentes substratos, de sementes recém-colhidas e armazenadas até 120 dias em laboratório, as quais foram coletadas no Pantanal do Rio Negro, Estado do Mato Grosso do Sul. Foram realizados testes com sementes recém-colhidas e após 30, 60, 90 e 120 dias de armazenamento, em três tipos de substrato (papel-filtro, solo arenoso e solo arenoso + matéria orgânica. Os resultados revelaram que C. glabrata possui pequena porcentagem de água (1,55% nas sementes, que decai com o período de armazenamento. Seu período de viabilidade é curto (90 dias e os melhores resultados foram obtidos com sementes recém-colhidas germinadas em papel-filtro (34%, com índice de velocidade de germinação de 10.Information about the feasibility and adequate substrates for the germination of seeds from native arboreous species are scarce. Cordia glabrata (louro preto, a pioneering species with wide distribution, is included among these species. Its characteristics allow its use in ornamentation, apiculture and popular medicine. Considering its potential of utilization, this work had as objective to evaluate the germination rates, in different substrates, of recently reaped seeds stored for up to 120 days in laboratory, which were collected in Pantanal of the Rio Negro, Mato Grosso do Sul State. Tests were done with recently reaped seeds and after 30, 60, 90 and 120 days of storage, in three kinds of substrate (paper filter, sandy soil and sandy soil + organic matter. The results revealed that C

  6. Plasma thromboxane B2 levels in horses experimentally infected with Strongylus vulgaris.

    Science.gov (United States)

    Cambridge, H; Reynoldson, J A; Dunsmore, J D

    1989-06-01

    Plasma thromboxane B2 (TXB2) the stable inactive metabolite of thromboxane A2 (TXA2), was measured daily by specific radioimmunoassay in three groups of animals before and after experimental infection with Strongylus vulgaris. Infection of four 'parasite naive' foals produced a typical acute syndrome with intermittent but statistically insignificant rises in TXB2 levels. Interpretation of results was complicated by the presence of a non-septic peritonitis associated with implantation of the foals with electrodes for recording myoelectrical activity. In two foals of similar age, with some natural exposure to S. vulgaris, there was little or no clinical response to infection and increases in TXB2 were absent. Baseline levels were also much lower, indicating that the peritonitis may have affected the results obtained in the first group of foals. Severe mesenteric arteritis was confirmed at necropsy in all six foals. A third group of yearling horses, all with natural exposure to the parasite, were generally resistant to infection. One animal developed arteritis with clinical signs of diarrhoea and mild colic, and also showed intermittent increases in TXB2. The mean plasma TXB2 level after infection was significantly higher than in the control period, although absolute levels were lower than those recorded in the 'parasite naive' foals. Other animals in this group had low TXB2 levels and minimal arteritis was found at necropsy. These results indicate that although infection appears to have an effect on plasma TXB2, the changes are inconsistent and not reliable indicators of the presence of verminous arteritis. The results also confirm the difficulty in establishing infection and the variability of the response in animals with previous exposure.

  7. Experimental infection of highly pathogenic avian influenza virus H5N1 in black-headed gulls (Chroicocephalus ridibundus)

    NARCIS (Netherlands)

    A. Ramis (Antonio); G. van Amerongen (Geert); M.W.G. van de Bildt (Marco); L.M.E. Leijten (Lonneke); R. Vanderstichel (R.); A.D.M.E. Osterhaus (Albert); T. Kuiken (Thijs)

    2014-01-01

    textabstractHistorically, highly pathogenic avian influenza viruses (HPAIV) rarely resulted in infection or clinical disease in wild birds. However, since 2002, disease and mortality from natural HPAIV H5N1 infection have been observed in wild birds including gulls. We performed an experimental

  8. [Experimental model of activated Lamblia (Giardia) muris infection in albino mice].

    Science.gov (United States)

    Irikov, O A; Kovalenko, F P

    2007-01-01

    Experimental L. muris infection was reproduced in 100% of the intact albino mice intragastrically given levomycin in an average total dose of 15.88-34.84 or 0.88-1.02 g/kg for 18-34 days. With levomycin administration, the intensity of giardiasis was 1121.6-8540.1 (mean 4830.9) thousand L. muris trophozoites per animal. The total number of trophozoites per animal decreased to 302.2-3481.4 (mean 1546.4) thousand and 28.1-324.0 (mean 109.4) thousand specimens 5-8 and 11-13 days after discontinuation of the antibiotic, respectively. The maximum number of L. muris trophozoites was observed in the proximal and middle portions of the murine small intestine during and after the administration oflevomycin. The highest isolation of cysts was seen 12-14 days after the initiation of administration of the antibiotic. Following 8-10 days of terminations of a course of levomycin therapy the native smear of animal feces showed no Lamblia cysts. In mice with activated infection, the isolation rate of Lamblia cysts was directly related to the intensity of intestinal infection with trophozoites of the parasite.

  9. Quasi-experimental Studies in the Fields of Infection Control and Antibiotic Resistance, Ten Years Later: A Systematic Review.

    Science.gov (United States)

    Alsaggaf, Rotana; O'Hara, Lyndsay M; Stafford, Kristen A; Leekha, Surbhi; Harris, Anthony D

    2018-02-01

    OBJECTIVE A systematic review of quasi-experimental studies in the field of infectious diseases was published in 2005. The aim of this study was to assess improvements in the design and reporting of quasi-experiments 10 years after the initial review. We also aimed to report the statistical methods used to analyze quasi-experimental data. DESIGN Systematic review of articles published from January 1, 2013, to December 31, 2014, in 4 major infectious disease journals. METHODS Quasi-experimental studies focused on infection control and antibiotic resistance were identified and classified based on 4 criteria: (1) type of quasi-experimental design used, (2) justification of the use of the design, (3) use of correct nomenclature to describe the design, and (4) statistical methods used. RESULTS Of 2,600 articles, 173 (7%) featured a quasi-experimental design, compared to 73 of 2,320 articles (3%) in the previous review (Pquasi-experimental design; and 68 (39%) identified their design using the correct nomenclature. In addition, 2-group statistical tests were used in 75 studies (43%); 58 studies (34%) used standard regression analysis; 18 (10%) used segmented regression analysis; 7 (4%) used standard time-series analysis; 5 (3%) used segmented time-series analysis; and 10 (6%) did not utilize statistical methods for comparisons. CONCLUSIONS While some progress occurred over the decade, it is crucial to continue improving the design and reporting of quasi-experimental studies in the fields of infection control and antibiotic resistance to better evaluate the effectiveness of important interventions. Infect Control Hosp Epidemiol 2018;39:170-176.

  10. E-NTPDase and E-ADA activities in rats experimental infected by Cryptococcus neoformans.

    Science.gov (United States)

    de Azevedo, Maria Isabel; Ferreiro, Laerte; Da Silva, Aleksandro S; Tonin, Alexandre A; Ruchel, Jader B; Rezer, João F P; França, Raqueli T; Zimmermann, Carine E P; Leal, Daniela B R; Duarte, Marta M M F; Lopes, Sonia T A; Flores, Mariana M; Fighera, Rafael; Santurio, Janio M

    2014-11-07

    Cryptococcus neoformans, the etiological agent of cryptococcosis, is an opportunistic fungal pathogen of immunocompromised individuals. The aim of this study was to evaluate the activities of E-NTPDase and E-ADA in rats experimentally infected by C. neoformans var. grubii. Adult rats (35) were divided in two groups: 18 for the control group (uninfected) (A), and 17 for the infected group (B). Each group was separated into three sub-groups (A1, A2, A3-B1, B2, B3), and samples were collected on 10, 20, and 30 days post-infection (PI). Leukocyte counts, IFN-γ, TNF-α, IgM, IgG levels, and E-NTPDase and E-ADA activities were analyzed. It was possible to observe that IgG and IgM seric levels of infected rats were significantly elevated (PADA activity had a significant reduction (PADA activity in lymphocytes increased significantly (PADA), concomitantly with an inflammatory response (increased levels of cytokines and immunoglobulins) associated with inflammatory infiltrates and histological lesions in the lung. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Ultrastructural identification of Ehrlich ia sp in an experimentally infected dog in Venezuela

    International Nuclear Information System (INIS)

    Gutierrez, N.; Martinez, M.; Arraga Alvarado, C.; Bretana, A.; Pacheco, I.; Comach, G.

    1999-01-01

    This study is the first report made in Venezuela concerning the ultrastructural characteristics of Ehrlich ia sp in mononuclear blood cells from an experimentally infected dog. The animal developed clinical manifestations characteristic of the infection, and typical intracytoplasmic inclusion bodies were clearly seen in blood smears stained with modified Giemsa examined by light microscopy. Microorganisms were visualized by transmission electron microscopy. The cytoplasmic inclusions, consisted of membrane-lined vacuole-containing elementary bodies. The organisms were extremely pleomorphic. Elementary bodies were surrounded by two distinct membranes and each was constituted by electro-dense granules. These findings corresponded to the described electron microscopy morphology which characterizes the Ehrlich ia genus

  12. Experimental infections with Mycoplasma agalactiae identify key factors involved in host-colonization.

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    Eric Baranowski

    Full Text Available Mechanisms underlying pathogenic processes in mycoplasma infections are poorly understood, mainly because of limited sequence similarities with classical, bacterial virulence factors. Recently, large-scale transposon mutagenesis in the ruminant pathogen Mycoplasma agalactiae identified the NIF locus, including nifS and nifU, as essential for mycoplasma growth in cell culture, while dispensable in axenic media. To evaluate the importance of this locus in vivo, the infectivity of two knock-out mutants was tested upon experimental infection in the natural host. In this model, the parental PG2 strain was able to establish a systemic infection in lactating ewes, colonizing various body sites such as lymph nodes and the mammary gland, even when inoculated at low doses. In these PG2-infected ewes, we observed over the course of infection (i the development of a specific antibody response and (ii dynamic changes in expression of M. agalactiae surface variable proteins (Vpma, with multiple Vpma profiles co-existing in the same animal. In contrast and despite a sensitive model, none of the knock-out mutants were able to survive and colonize the host. The extreme avirulent phenotype of the two mutants was further supported by the absence of an IgG response in inoculated animals. The exact role of the NIF locus remains to be elucidated but these data demonstrate that it plays a key role in the infectious process of M. agalactiae and most likely of other pathogenic mycoplasma species as many carry closely related homologs.

  13. Comparison of abortion and infection after experimental challenge of pregnant bison and cattle with Brucella abortus strain 2308

    Science.gov (United States)

    A comparative study was conducted using data from naive bison (n=45) and cattle (n=46) from 8 and 6 studies, respectively, in which a standardized Brucella abortus strain 2308 experimental challenge was administered. The incidence of abortion, fetal infection, uterine or mammary infection, or infec...

  14. Cross-Species Infectivity of H3N8 Influenza Virus in an Experimental Infection in Swine.

    Science.gov (United States)

    Solórzano, Alicia; Foni, Emanuela; Córdoba, Lorena; Baratelli, Massimiliano; Razzuoli, Elisabetta; Bilato, Dania; Martín del Burgo, María Ángeles; Perlin, David S; Martínez, Jorge; Martínez-Orellana, Pamela; Fraile, Lorenzo; Chiapponi, Chiara; Amadori, Massimo; del Real, Gustavo; Montoya, María

    2015-11-01

    Avian influenza A viruses have gained increasing attention due to their ability to cross the species barrier and cause severe disease in humans and other mammal species as pigs. H3 and particularly H3N8 viruses, are highly adaptive since they are found in multiple avian and mammal hosts. H3N8 viruses have not been isolated yet from humans; however, a recent report showed that equine influenza A viruses (IAVs) can be isolated from pigs, although an established infection has not been observed thus far in this host. To gain insight into the possibility of H3N8 avian IAVs to cross the species barrier into pigs, in vitro experiments and an experimental infection in pigs with four H3N8 viruses from different origins (equine, canine, avian, and seal) were performed. As a positive control, an H3N2 swine influenza virus A was used. Although equine and canine viruses hardly replicated in the respiratory systems of pigs, avian and seal viruses replicated substantially and caused detectable lesions in inoculated pigs without previous adaptation. Interestingly, antibodies against hemagglutinin could not be detected after infection by hemagglutination inhibition (HAI) test with avian and seal viruses. This phenomenon was observed not only in pigs but also in mice immunized with the same virus strains. Our data indicated that H3N8 IAVs from wild aquatic birds have the potential to cross the species barrier and establish successful infections in pigs that might spread unnoticed using the HAI test as diagnostic tool. Although natural infection of humans with an avian H3N8 influenza A virus has not yet been reported, this influenza A virus subtype has already crossed the species barrier. Therefore, we have examined the potential of H3N8 from canine, equine, avian, and seal origin to productively infect pigs. Our results demonstrated that avian and seal viruses replicated substantially and caused detectable lesions in inoculated pigs without previous adaptation. Surprisingly, we

  15. Inactivation of Candida glabrata by a humid DC argon discharge afterglow: dominant contributions of short-lived aqueous active species

    International Nuclear Information System (INIS)

    Xiong, Qing; Liu, Hongbin; Xu, Le; Wang, Xia; Zhu, Qunlin; Lu, Weiping; Chen, Qiang; Zeng, Xue; Yi, Ping

    2017-01-01

    Plasma medicine applications are currently attracting significant interest all over the world. Bactericidal treatments of Candida glabrata cultured in saline suspension are performed in this study by a room-temperature reactive afterglow of a DC-driven argon discharge. Water vapor was added to the discharge to study the inactivation contributions of reactive hydrolytic species including OH and H 2 O 2 transporting along the gas flow to the treated solutions. The inactivation results indicate that the dominant roles in the bactericidal treatments are played by the short-lived aqueous active species, but not the stable species like H 2 O 2aq (aq indicates an aqueous species). Further analysis shows that the ·OH aq radicals play an important role in the inactivation process. The ·OH aq radicals in the suspension are mostly produced from the direct dissolution of the OH species in the reactive afterglow. With the increase of added water vapor content, the ·OH aq production increases and enhances the inactivation efficiency of C. glabrata . Furthermore, it is found that the ambient air diffusion shows essential effects on the bactericidal activity of the remote humid argon discharge. Higher bactericidal effects can be obtained in open-space treatments compared to in a controlled Ar + H 2 O gas atmosphere. Key active air-byproduct species are believed to be generated in the suspension during the treatments and contributing to the inactivation process. Based on chemical analysis, the peroxynitrous acid ONOOH aq is considered as the key antimicrobial air-byproduct species. These results indicate the important dependence of plasma biomedical effects on the processing environment, which finally relates to the critical contributions of the key reactive species formed therein. (paper)

  16. [INTESTINAL FAILURE AND YERSINIA PSEUDOTUBERCULOSIS TRANSLOCATION IN THE DEVELOPMENTOF EXPERIMENTAL GENERALIZED INFECTION].

    Science.gov (United States)

    Chicherin, I Yu; Pogorelky, I P; Lundovskikh, I A; Darmov, I V; Gorshkov, A S; Shabalina, M R

    2016-01-01

    To determine the value of intestinal failure and translocation of bacteria Y. pseudotuberculosis, and normal intestinal microbiota in the initiation and generalization of infection in experimental pseudotuberculosis in conventional white mice, as well as pathological manifestation of it as a response to the adhesion and colonization of the mucosus membrane by pathogenic bacteria Y. pseudotuberculosis. Experimental models of pseudotuberculosis in conventional white mice used the pathogenic Y. pseudotuberculosis 147 serotype I strain, containing a calcium-dependence plasmid with a molecular weight of 47 MDa. Cultivation of the pseudotuberculosis pathogen given its psychrophilic was performed on Hottinger agar at a temperature of (4-5) °C. The lactobacilli strain L plantarum 8P-A3 was isolated from a lyophilized commercial probiotic Lactobacterin (manufactured by "NPO Microgen", Russia) and used to obtain native culture supernatant fluid of lactobacilli, the composition of which was detected by gas-liquid chromatography with mass-selective detection. Gentamicin for parenteral administration was manufactured by JSC "Biochemist", Russia. Pathomorphological examination was performed on the 4-6th day of the experiment. Fragments of the small intestine, liver, kidneys, and lungs from dead animals were chosen for examination. Tissues were fixed in 10% neutral formalin, dehydrated in isopropanol and embedded in paraffin. Preparations were stained with Ehrlich hematoxylin and eosin, examined on the microscope "Mikmed-2" (JSC "LOMO", Russia) under magnification x 200-x1000. Statistical processing of the experimental results was carried out according to the method of Kerber in modification of I.P. Ashmarin and A.A. Vorobyov. The role of intestinal failure and translocation of bacteria Y. pseudotuberculosis, and normal intestinal microbiota in the initiation and generalization of infection in animals has been found. It has been proved that the oral administration of supernatant

  17. Studies on survival, biological activities and behavior of Biomphalaria glabrata, the host snail of Schistosomiasis, submitted to increased hydrostatic pressure: a technique Estudos sobre a sobrevivência, atividades biológicas e comportamento de Biomphalaria glabrata, hospedeiro intermediário da esquistossomose, submetido a aumento da pressão hidrostática: uma técnica

    Directory of Open Access Journals (Sweden)

    P. Jurberg

    1988-03-01

    Full Text Available To study changes in survival, in biological activities and behavior of planorbids submitted to increased hydrostatic pressure, we developed a technique using two transparent chambers and a hydraulic piston. The apparatus permitted renewal of the liquid medium without substantial variations in pressure, thus eliminating excretion products and maintaining the desired O2 level and thereby permitting us to evaluate the effects of pressure independently of the occurrence of anoxia. Pressure was maintained without any contact of the liquid medium with compressed air, a situation which reproduced with relative fidelity what occurs in nature and assured the presence of the same amounts of gases in the two observation chambers (Control and Experimental. Biomphalaria glabrata was found to be able to survive at least 48 hours when submitted to 49.02 x 10**4 Pa (equivalent to a water depth of 48.8 m, continuing to day egg masses and showing few behavioral changes when compared with the control group.Para estudar mudanças na sobrevivência, atividades biológicas e comportamento de planorbídeos submetidos a aumento de pressão hidrostática, desenvolvemos uma técnica que utiliza duas câmaras transparentes e um pistão hidráulico. O aparelho permitiu a renovação do meio líquido sem variações substanciais na pressão, eliminando assim os produtos de excreção e mantendo o nível de O2 disseolvido desejado, e desse modo permitido-nos avaliar o efeito da pressão independente da ocorrência de anoxia. A pressão foi mantida sem nenhum contato do meio líquido com o ar comprimido, situação que produziu com relativa fidelildade o que ocorre na natureza, e assegurou a presença da mesma quantidade de gases nas duas câmaras de observação. Biomphalaria glabrata foi capaz de sobreviver pelo menos 48 horas quando submetida a 49,02 x 10 Pa (equivalente a 48 m de profundidade continuando a pôr massas ovíferas, e mostrando poucas modifica

  18. Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice

    Directory of Open Access Journals (Sweden)

    Jamile R. Silva

    2016-01-01

    Full Text Available Ureaplasma diversum is an opportunistic pathogen associated with uterine inflammation, impaired embryo implantation, infertility, abortions, premature birth of calves and neonatal pneumonia in cattle. It has been suggested that the intra-uterine infection by Ureaplasma diversum can cause vascular changes that hinder the success of pregnancy. Thus, the aim of this study was to evaluate the changes of intrauterine site of A/J mice in estrus or proestrus phase inoculated with Ureaplasma diversum. The infection was monitored at 24, 48 and 72 hours by the PCR methodology to detect the Ureaplasma in the inoculation site and the profile of circulating blood cells. Morphological changes, intensity of inflammation and the production of cytokines were compared. The infected mice showed local inflammation through the production of IFN-γ and TNF-α. Ureaplasma diversum infections in the reproductive tract of studied mice seemed to be associated with the production of pro-inflammatory cytokines in uterine parenchyma. The levels of TNF-α of infected mice were dependent on the bacterial load of inoculated Ureaplasma. Uterine experimental infections by Ureaplasma diversum have not been mentioned yet and herein we presented the first report of an intrauterine infection model in mice.

  19. Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice.

    Science.gov (United States)

    Silva, Jamile R; Ferreira, Lício F A A; Oliveira, Percíllia V S; Nunes, Ivanéia V; Pereira, Ítalo S; Timenetsky, Jorge; Marques, Lucas M; Figueiredo, Tiana B; Silva, Robson A A

    2016-01-01

    Ureaplasma diversum is an opportunistic pathogen associated with uterine inflammation, impaired embryo implantation, infertility, abortions, premature birth of calves and neonatal pneumonia in cattle. It has been suggested that the intra-uterine infection by Ureaplasma diversum can cause vascular changes that hinder the success of pregnancy. Thus, the aim of this study was to evaluate the changes of intrauterine site of A/J mice in estrus or proestrus phase inoculated with Ureaplasma diversum. The infection was monitored at 24, 48 and 72 hours by the PCR methodology to detect the Ureaplasma in the inoculation site and the profile of circulating blood cells. Morphological changes, intensity of inflammation and the production of cytokines were compared. The infected mice showed local inflammation through the production of IFN-γ and TNF-α. Ureaplasma diversum infections in the reproductive tract of studied mice seemed to be associated with the production of pro-inflammatory cytokines in uterine parenchyma. The levels of TNF-α of infected mice were dependent on the bacterial load of inoculated Ureaplasma. Uterine experimental infections by Ureaplasma diversum have not been mentioned yet and herein we presented the first report of an intrauterine infection model in mice.

  20. Metabolomic profiling of faecal extracts from Cryptosporidium parvum infection in experimental mouse models.

    Directory of Open Access Journals (Sweden)

    Josephine S Y Ng Hublin

    Full Text Available Cryptosporidiosis is a gastrointestinal disease in humans and animals caused by infection with the protozoan parasite Cryptosporidium. In healthy individuals, the disease manifests mainly as acute self-limiting diarrhoea, but may be chronic and life threatening for those with compromised immune systems. Control and treatment of the disease is challenged by the lack of sensitive diagnostic tools and broad-spectrum chemotherapy. Metabolomics, or metabolite profiling, is an emerging field of study, which enables characterisation of the end products of regulatory processes in a biological system. Analysis of changes in metabolite patterns reflects changes in biochemical regulation, production and control, and may contribute to understanding the effects of Cryptosporidium infection in the host environment. In the present study, metabolomic analysis of faecal samples from experimentally infected mice was carried out to assess metabolite profiles pertaining to the infection. Gas-chromatography mass spectrometry (GC-MS carried out on faecal samples from a group of C. parvum infected mice and a group of uninfected control mice detected a mean total of 220 compounds. Multivariate analyses showed distinct differences between the profiles of C. parvum infected mice and uninfected control mice,identifying a total of 40 compounds, or metabolites that contributed most to the variance between the two groups. These metabolites consisted of amino acids (n = 17, carbohydrates (n = 8, lipids (n = 7, organic acids (n = 3 and other various metabolites (n = 5, which showed significant differences in levels of metabolite abundance between the infected and uninfected mice groups (p < 0.05. The metabolites detected in this study as well as the differences in abundance between the C. parvum infected and the uninfected control mice, highlights the effects of the infection on intestinal permeability and the fate of the metabolites as a result of nutrient scavenging by the

  1. Experimental infection of duck origin virulent Newcastle disease virus strain in ducks.

    Science.gov (United States)

    Dai, Yabin; Cheng, Xu; Liu, Mei; Shen, Xinyue; Li, Jianmei; Yu, Shengqing; Zou, Jianmin; Ding, Chan

    2014-07-17

    Newcastle disease (ND) caused by virulent Newcastle disease virus (NDV) is an acute, highly contagious and fatal viral disease affecting most species of birds. Ducks are generally considered to be natural reservoirs or carriers of NDV while being resistant to NDV strains, even those most virulent for chickens; however, natural ND cases in ducks have been gradually increasing in recent years. In the present study, ducks of different breeds and ages were experimentally infected with duck origin virulent NDV strain duck/Jiangsu/JSD0812/2008 (JSD0812) by various routes to investigate the pathogenicity of NDV in ducks. Six breeds (mallard, Gaoyou, Shaoxing, Jinding, Shanma, and Pekin ducks) were infected intramuscularly (IM) with JSD0812 strain at the dose of 5 × 108 ELD50. Susceptibility to NDV infection among breeds varied, per morbidity and mortality. Mallard ducks were the most susceptible, and Pekin ducks the most resistant. Fifteen-, 30-, 45-, 60-, and 110-day-old Gaoyou ducks were infected with JSD0812 strain at the dose of 5 × 108 ELD50 either IM or intranasally (IN) and intraocularly (IO), and their disease development, viral shedding, and virus tissue distribution were determined. The susceptibility of ducks to NDV infection decreased with age. Most deaths occurred in 15- and 30-day-old ducklings infected IM. Ducks infected IN and IO sometimes exhibited clinical signs, but seldom died. Clinical signs were primarily neurologic. Infected ducks could excrete infectious virus from the pharynx and/or cloaca for a short period, which varied with bird age or inoculation route; the longest period was about 7 days. The rate of virus isolation in tissues from infected ducks was generally low, even in those from dead birds, and it appeared to be unrelated to bird age and infection route. The results confirmed that some of the naturally occurring NDV virulent strains can cause the disease in ducks, and that ducks play an important role in the epidemiology of ND. The

  2. Infectivity of DWV associated to flower pollen: experimental evidence of a horizontal transmission route.

    Directory of Open Access Journals (Sweden)

    Maurizio Mazzei

    Full Text Available Deformed wing virus (DWV is a honeybee pathogen whose presence is generally associated with infestation of the colony by the mite Varroa destructor, leading to the onset of infections responsible for the collapse of the bee colony. DWV contaminates bee products such as royal jelly, bee-bread and honey stored within the infected hive. Outside the hive, DWV has been found in pollen loads collected directly from infected as well as uninfected forager bees. It has been shown that the introduction of virus-contaminated pollen into a DWV-free hive results in the production of virus-contaminated food, whose role in the development of infected bees from virus-free eggs has been experimentally demonstrated. The aim of this study was twofold: (i to ascertain the presence of DWV on pollen collected directly from flowers visited by honeybees and then quantify the viral load and (ii determine whether the virus associated with pollen is infective. The results of our investigation provide evidence that DWV is present on pollen sampled directly from visited flowers and that, following injection in individuals belonging to the pollinator species Apis mellifera, it is able to establish an active infection, as indicated by the presence of replicating virus in the head of the injected bees. We also provide the first indication that the pollinator species Osmia cornuta is susceptible to DWV infection.

  3. Testing the sensitivity of Nested PCR method to detect Aspergillus fumigates in experimentally infected Sputum samples

    International Nuclear Information System (INIS)

    Ramadan, A.; Soukkaria, S.

    2013-01-01

    Fungal infections caused by Aspergillus species generally are occupying a second place among invasive fungal infections in the world, especially A. fumigatus, which is considered the main cause of invasive Aspergillosis (IA). Although IA rarely infects immunocompetent individuals, however, it can lead to death in immunocompromised patients. Therefore, it is necessary to diagnose the infection early in order to treat the disease efficiently. However, the conventional diagnostic tools, currently used to detect infections, has low sensitivity and reliability. Polymerase chain reaction (PCR) technology distribution as a molecular and high sensitive technology has allowed us to make comparative study between sensitivity of traditional currently used diagnostic method and Nested-PCR, the result of the study of sputum samples that experimentally infected with different concentrations of A.fumigatus spores ramping from 10 to10 6 spore/ml, have high sensitivity and specificity of Nested-PCR in detecting the lower concentrations, comparing with traditional diagnostic method (culture on Sabouraud media) that were negative in all concetrations. (author)

  4. Studies on vertical transmission of Trichinella spiralis in experimentally infected guinea pigs (Cavia porcellus).

    Science.gov (United States)

    Riva, Eliana; Fiel, Cesar; Bernat, Gisele; Muchiut, Sebastián; Steffan, Pedro

    2017-08-01

    An experimental study to enhance knowledge on the capability of Trichenella spiralis to pass from guinea pigs to progeny at different periods of pregnancy or lactation was performed. For this purpose, 18 female adult guinea pigs were inoculated with 100 or 1000 T. spiralis muscle larvae (ML) during early, late gestation and during lactation period. The presence of T. spiralis (ML) in mothers and newborns was studied through enzymatic digestion from muscle samples. ML were observed in 9 of 42 newborn guinea pigs and levels of infection were significantly higher when infections of mothers were done during late gestation (p = 0.0046) with the high infective dose (p = 0.0043). T. spiralis ML were not recovered from any of the newborns from mothers infected in the lactation period. Ten out of 18 infected mothers presented larvae 1 in their mammary glands. Muscle samples from the tongue and the masseter showed the highest larval burdens. These observations confirm previous reports on that ML of T. spiralis are capable to pass through placental tissues to reach and encyst in striated muscle groups of newborn guinea pigs. This study may also reinforce the importance of preventive programs to control trichinellosis in those endemic areas where pregnant women would have high risk of infection.

  5. Pathology, clinical signs, and tissue distribution of Toxoplasma gondii in experimentally infected reindeer (Rangifer tarandus

    Directory of Open Access Journals (Sweden)

    Émilie Bouchard

    2017-12-01

    Full Text Available Toxoplasma gondii is a zoonotic parasite found in vertebrates worldwide for which felids serve as definitive hosts. Despite low densities of felids in northern Canada, Inuit people in some regions show unexpectedly high levels of exposure, possibly through handling and consumption of Arctic wildlife. Free-ranging caribou (Rangifer tarandus are widely harvested for food across the Canadian North, show evidence of seroexposure to T. gondii, and are currently declining in numbers throughout the Arctic. We experimentally infected three captive reindeer (conspecific with caribou with 1000, 5000 or 10,000 oocysts of T. gondii via stomach intubation to assess clinical signs of infection, pathology, and tissue distribution. An unexposed reindeer served as a negative control. Signs of stress, aggression, and depression were noted for the first two weeks following infection. By 4 weeks post infection, all infected reindeer were positive on a modified agglutination test at the highest titer tested (1:200 for antibodies to T. gondii. At 20 weeks post infection, no gross abnormalities were observed on necropsy. Following histopathology and immunohistochemistry, tissue cysts were visualized in the reindeer given the highest and lowest dose of oocysts. Focal pleuritis and alveolitis were associated with respiratory problems in reindeer given the middle dose. DNA of T. gondii was detected following traditional DNA extraction and conventional PCR on 25 mg samples from 17/33 muscles and organs, and by magnetic capture DNA extraction from 100 g samples from all 26 tissues examined. This research demonstrated that reindeer/caribou can serve as intermediate hosts for T. gondii, and that the parasite may be associated with health effects in wildlife. The presence of T. gondii in all tissues tested, many of which are commonly consumed raw, smoked, or dried in northern communities, suggests that caribou may serve as a source of human exposure to T

  6. Distribution of yeast species associated with oral lesions in HIV-infected patients in Southwest Uganda.

    LENUS (Irish Health Repository)

    Agwu, Ezera

    2012-04-01

    Oropharyngeal candidiasis remains a significant clinical problem in HIV-infected and AIDS patients in regions of Africa where anti-retroviral therapy isn\\'t readily available. In this study we identified the yeast populations associated with oral lesions in HIV-infected patients in Southwest Uganda who were receiving treatment with nystatin and topical clotrimazole. Samples were taken from 605 patients and 316 (52%) of these yielded yeast growth following incubation on Sabouraud dextrose agar. Samples were subsequently re-plated on CHROMagar Candida medium to facilitate identification of the yeast species present. The majority (56%) of culture-positive samples yielded a mix of two or more species. Candida albicans was present in 87% (274\\/316) of patient samples and accounted for 87% (120\\/138) of single species samples. Candida glabrata, Candida tropicalis and Candida norvegensis were also found in cultures that yielded a single species. No Candida dubliniensis isolates were identified in this population.

  7. Distribution of yeast species associated with oral lesions in HIV-infected patients in Southwest Uganda.

    Science.gov (United States)

    Agwu, Ezera; Ihongbe, John C; McManus, Brenda A; Moran, Gary P; Coleman, David C; Sullivan, Derek J

    2012-04-01

    Oropharyngeal candidiasis remains a significant clinical problem in HIV-infected and AIDS patients in regions of Africa where anti-retroviral therapy isn't readily available. In this study we identified the yeast populations associated with oral lesions in HIV-infected patients in Southwest Uganda who were receiving treatment with nystatin and topical clotrimazole. Samples were taken from 605 patients and 316 (52%) of these yielded yeast growth following incubation on Sabouraud dextrose agar. Samples were subsequently re-plated on CHROMagar Candida medium to facilitate identification of the yeast species present. The majority (56%) of culture-positive samples yielded a mix of two or more species. Candida albicans was present in 87% (274/316) of patient samples and accounted for 87% (120/138) of single species samples. Candida glabrata, Candida tropicalis and Candida norvegensis were also found in cultures that yielded a single species. No Candida dubliniensis isolates were identified in this population.

  8. Immunohistochemical detection of Tritrichomonas foetus in experimentally infected mice

    Directory of Open Access Journals (Sweden)

    Monteavaro Cristina Esther

    2000-01-01

    Full Text Available The need to intensify knowledge of the pathogenesis of bovine genital trichomoniasis (BGT led to the use of alternative animal models such as the mouse. Nevertheless, it is necessary to elucidate the dynamics of the infection in this animal species, evaluating different stages of the colonization and evolution of the pathological alterations. The immunohistochemistry (IHC offers advantages over the routine histopathological staining techniques for the detection of the protozoan in tissues, cellular detritus and inside the macrophages. The goal of the present study was to demonstrate the presence of Tritrichomonas foetus in the reproductive tract of infected mice using an IHC technique. Female BALB/c mice were infected with a suspension of T. foetus by intravaginal route, in the estrum phase, detected by exfoliative vaginal cytology. After 10 weeks, the animals were sacrificed; uterus and vagina were fixed and histologically processed. Some slides were stained with HE. The rest of the slides were processed for IHC. An immunoadsorbed polyclonal serum against T. foetus was used. The avidine-biotine technique (HistoMouse, Zymed[tm] was employed. The histopathological studies showed a dilation of the uterine glands, presence of macrophages in the lumen of the organ and inner part of the endometrial glands. No T. foetus was identified using this method. The IHQ allowed additionally the identification of the protozoan in the endometrium, endometrial glands, uterine lumen and inside neutrophils and macrophages. The cytological studies stained with IHC showed either isolated T. foetus adhered to epithelial cells or inside macrophages. This technique proves to be a useful tool for the study of the pathogenesis of bovine genital trichomoniasis (BGT in an experimental model.

  9. Experimentally induced intravaginal Tritrichomonas foetus infection in a mouse model Infecção experimental intravaginal com Tritricho-monas foetus em modelo camundongo

    Directory of Open Access Journals (Sweden)

    Pedro Soto

    2005-12-01

    Full Text Available The interest to develop research on the host-parasite relationship in bovine tritrichomonosis has accomplished the use of experimental models alternative to cattle. The BALB/c mouse became the most appropriate species susceptible to vaginal Tritrichomonas foetus infection requiring previous estrogenization. For the need of an experimental model without persistent estrogenization and with normal estrous cycles, the establishment and persistence of vaginal infection on BALB/c mouse with different concentrations of T. foetus in two experimental groups was evaluated. Group A was treated with 5mg of b-estradiol 3-benzoate to synchronize the estrous, 48 hours before the T. foetus vaginal inoculation, and Group B was inoculated in natural estrus. At 5-7 days after treatment, estrogenic effect decreased allowing all animals to cycle regularly during the experiment. From the first week post-infection, samples of vaginal mucus were taken from all animals during 34 weeks, in order to evaluate the course of infection and the stage of the estrus cycle. Group A showed 93.6% of infected animals, and Group B showed 38%. Different doses of T. foetus were assayed to establish the vaginal infection, with a persistence of 34 weeks. Although different behavior was observed in each subgroup belonging to either Group A or Group B, there were no significant differences among the infecting doses used. The b-estradiol 3-benzoate treatment had a favorable effect on the establishment of the infection (PA necessidade de esclarecer a relação agente-hospedeiro na tricomoníase bovina deu motivo para o uso de modelos experimentais alternativos ao bovino. O camundongo BALB/c resultou como espécie mais adequada para a infeção vaginal com Tritrichomonas foetus, requerendo uma estrogenização prévia. Visando a necessidade de um modelo experimental sem estrogenização persistente e com ciclos estrais normais, foi avaliada a instalação e persistência da infeção vaginal

  10. Experimental infection of pigs with two East European variants of Type 1 PRRSV

    DEFF Research Database (Denmark)

    Hjulsager, Charlotte Kristiane; Larsen, Lars Erik; Heegaard, Peter M. H.

    Porcine reproductive and respiratory syndrome viruses (PRRSV) have been divided into Type 1 (European) and Type 2 (North American) viruses. PRRSV are very diverse and Type 1 viruses have even been further divided into subtypes. While Type 1 viruses from Western Europe belong to subtype 1, viruses...... the subtype 1 strains. The aim of this project was to study the infection dynamics and clinical and pathological impact of two east European Type 1 strains. In an experimental trial, infection of pigs with the Russian subtype 2 strain “Ili6” and the Belarusian atypical isolate “Bor59” were compared...... to an early “Lelystad-like” Danish subtype 1 isolate “18794”. Groups of seven pigs of unique high sanitary status were infected with one of the three PRRSV isolates, and a fourth group served as sham-inoculated controls. The pigs were monitored for 24 days, and nasal swabs and blood samples were taken at 0, 3...

  11. An investigation on vertical transmission of Leishmania infantum in experimentally infected dogs and assessment of offspring's infectiousness potential by xenodiagnosis.

    Science.gov (United States)

    Ben Slimane, T; Chouihi, E; Ben Hadj Ahmed, S; Chelbi, I; Barhoumi, W; Cherni, S; Zoghlami, Z; Gharbi, M; Zhioua, E

    2014-12-15

    Dogs are the main reservoir host of Leishmania infantum, etiologic agent of human visceral leishmaniasis (HVL) and canine visceral leishmaniasis (CVL). Transmission of L. infantum to humans and dogs is mainly through the bite of infected sand flies. In the Western Mediterranean basin, Phlebotomus perniciosus is the main vector of L. infantum. However, occasional vertical transmission of L. infantum has been reported. This study investigated L. infantum vertical transmission in offspring of experimentally infected dogs. Among 14 surviving puppies from three female beagle dogs that developed CVL following an experimental infection with L. infantum, one was tested positive by indirect immunofluorescence antibody test, by PCR and by xenodiagnosis with a high parasite burden in the spleen at 14 months old. None of the remaining puppies were tested positive for L. infantum. These findings strongly suggest that infected puppies following vertical transmission can sustain infection and contribute in infecting sand flies with L. infantum. Any strategy for controlling CVL should take into consideration the vertical transmission of L. infantum. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Experimental infection of Marmota monax with a novel hepatitis A virus.

    Science.gov (United States)

    Yu, Jie-Mei; Li, Li-Li; Xie, Guang-Cheng; Zhang, Cui-Yuan; Ao, Yuan-Yun; Duan, Zhao-Jun

    2018-05-01

    To establish an animal model for the newly identified Marmota Himalayana hepatovirus, MHHAV, so as to develop a better understanding of the infection of hepatitis A viruses. Five experimental woodchucks (Marmota monax) were inoculated intravenously with the purified MHHAV from wild woodchuck feces. One animal injected with PBS was defined as a control. Feces and blood were routinely collected. After the animals were subjected to necropsy, different tissues were collected. The presence of viral RNA and negative sense viral RNA was analyzed in all the samples and histopathological and in situ hybridization analysis was performed for the tissues. MHHAV infection caused fever but no severe symptoms or death. Virus was shed in feces beginning at 2 dpi, and MHHAV RNA persisted in feces for ~2 months, with a biphasic increase, and in blood for ~30 days. Viral RNA was detected in all the tissues, with high levels in the liver and spleen. Negative-strand viral RNA was detected only in the liver. Furthermore, the animals showed histological signs of hepatitis at 45 dpi. MHHAV can infect M. monax and is associated with hepatic disease. Therefore, this animal can be used as a model of HAV pathogenesis and to evaluate antiviral and anticancer therapeutics.

  13. Aeromonas caviae inhibits hepatic enzymes of the phosphotransfer network in experimentally infected silver catfish: Impairment on bioenergetics.

    Science.gov (United States)

    Baldissera, M D; Souza, C F; Verdi, C M; Dos Santos, K L M; Da Veiga, M L; da Rocha, M I U M; Santos, R C V; Vizzotto, B S; Baldisserotto, B

    2018-03-01

    Several studies have been demonstrated that phosphotransfer network, through the adenylate kinase (AK) and pyruvate kinase (PK) activities, allows for new perspectives leading to understanding of disease conditions associated with disturbances in energy metabolism, metabolic monitoring and signalling. In this sense, the aim of this study was to evaluate whether experimental infection by Aeromonas caviae alters hepatic AK and PK activities of silver catfish Rhamdia quelen. Hepatic AK and PK activities decreased in infected animals compared to uninfected animals, as well as the hepatic adenosine triphosphate (ATP) levels. Also, a severe hepatic damage was observed in the infected animals due to the presence of dilation and congestion of vessels, degeneration of hepatocytes and loss of liver parenchyma architecture and sinusoidal structure. Therefore, we have demonstrated, for the first time, that experimental infection by A. caviae inhibits key enzymes linked to the communication between sites of ATP generation and ATP utilization. Moreover, the absence of a reciprocal compensatory mechanism between these enzymes contributes directly to hepatic damage and for a severe energetic imbalance, which may contribute to disease pathophysiology. © 2017 John Wiley & Sons Ltd.

  14. Fibrinogen and fibronectin binding cooperate for valve infection and invasion in Staphylococcus aureus experimental endocarditis

    NARCIS (Netherlands)

    Que, Yok-Ai; Haefliger, Jacques-Antoine; Piroth, Lionel; François, Patrice; Widmer, Eleonora; Entenza, José M; Sinha, Bhanu; Herrmann, Mathias; Francioli, Patrick; Vaudaux, Pierre; Moreillon, Philippe

    2005-01-01

    The expression of Staphylococcus aureus adhesins in Lactococcus lactis identified clumping factor A (ClfA) and fibronectin-binding protein A (FnBPA) as critical for valve colonization in rats with experimental endocarditis. This study further analyzed their role in disease evolution. Infected

  15. Experimental Maedi Visna Virus Infection in sheep: a morphological, immunohistochemical and PCR study after three years of infection

    Directory of Open Access Journals (Sweden)

    S Preziuso

    2009-06-01

    Full Text Available A morphological, immunohistochemical and polymerase chain reaction (PCR study was performed on eight ewes experimentally infected with an Italian strain of Maedi-Visna Virus (MVV in order to evaluate the lesions and the viral distribution after three years of infection. At the moment of euthanasia, seven sheep were seropositive for MVV, while one sheep in poor body conditions was seronegative since one year. Lungs, pulmonary lymph nodes, udder, supramammary lymph nodes, carpal joints, the CNS, spleen and bone marrow of the eight infected sheep were collected for histology, for immunohistochemical detection of the MVV core protein p28 and for PCR amplification of a 218 bp viral DNA sequence of the pol region. The most common histological findings consisted of interstitial lymphoproliferative pneumonia and lymphoproliferative mastitis of different severity, while no lesions were observed in the CNS. MVV p28 antigen was immunohistochemically labelled in lungs, udder, pulmonary lymph nodes, spleen and bone marrow but not in the CNS of all the eight infected sheep. A 218 bp sequence of MVV pol region was detected in lung of a seropositive and of the seroconverted negative sheep. The results suggest that (i MVV causes heterogeneous lesions in homogeneously reared ewes, (ii MVV p28 antigen is detectable not only in inflammed target organs, but also in pulmonary lymph nodes, spleen and bone marrow, and (iii immunohistochemistry and PCR are useful methods for Maedi-Visna diagnosis in suspected cases, also when serological tests are negative.

  16. Biological control of schistosome transmission in flowing water habitats.

    Science.gov (United States)

    Jobin, W R; Laracuente, A

    1979-09-01

    Marisa cornuarietis was evaluated in Puerto Rico for control of schistosome transmission in flowing water. A population of Biomphalaria glabrata and their schistosome infections disappeared after introduction of 20,000 M. cornuarietis to an endemic stream, while in nearby untreated streams the B. glabrata population remained stable and the schistosome prevalence increased. This method cost U.S. $0.10 per capita for over a year of protection, 5%-10% of the cost of chemical control.

  17. Experimental infection with Escherichia coli 0149 : F4ac in weaned piglets

    DEFF Research Database (Denmark)

    Jensen, Gerda M.; Frydendahl, Kai; Svendsen, Ove

    2006-01-01

    adhesion test made after slaughter of piglets. However, in an experimental infection study with the purpose to obtain diarrhoeic piglets, it would be an advantage to test for susceptibility prior to experimentation. The Mucin 4 gene on porcine chromosome 13 has been proposed as a candidate gene...... for the production of the specific ETEC F4ab/ac receptor, and a DNA marker-based test has been developed to allow genotyping for ETEC F4ab/ac resistance/susceptibility [Jorgensen, C.B., Cirera, S., Archibald, A.L., Anderson, L., Fredholm, M., Edfors-Lilja, I., 2004. Porcine polymorphisms and methods for detecting...... them. International application published under the patent cooperation treaty (PCT). PCT/DK2003/000807 or WO2004/048606-A2]. The aim of this study was to test an experimental model for ETEC O149:F4ac-induced diarrhoea in piglets, selected for susceptibility towards ETEC O149:F4ac adhesion prior...

  18. Detection and distribution of ostreid herpesvirus 1 in experimentally infected Pacific oyster spat.

    Science.gov (United States)

    Segarra, Amélie; Baillon, Laury; Faury, Nicole; Tourbiez, Delphine; Renault, Tristan

    2016-01-01

    High mortality rates are reported in spat and larvae of Pacific oyster Crassostrea gigas and associated with ostreid herpesvirus 1 (OsHV-1) detection in France. Although the viral infection has been experimentally reproduced in oyster larvae and spat, little knowledge is currently available concerning the viral entry and its distribution in organs and tissues. This study compares OsHV-1 DNA and RNA detection and localization in experimentally infected oysters using two virus doses: a low dose that did not induce any mortality and a high dose inducing high mortality. Real time PCR demonstrated significant differences in terms of viral DNA amounts between the two virus doses. RNA transcripts were detected in oysters receiving the highest dose of viral suspension whereas no transcript was observed in oysters injected with the low dose. This study also allowed observing kinetics of viral DNA and RNA detection in different tissues of oyster spat. Finally, viral detection was significantly different in function of tissues (p<0.005), time (p<0.005) with an interaction between tissues and time (p<0.005) for each probe. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Cranberry juice and combinations of its organic acids are effective against experimental urinary tract infection

    DEFF Research Database (Denmark)

    Jensen, Heidi Dorthe; Struve, Carsten; Christensen, Søren Brøgger

    2017-01-01

    The antibacterial effect of cranberry juice and the organic acids therein on infection by uro28 pathogenic Escherichia coli was studied in an experimental mouse model of urinary tract infection (UTI). Reduced bacterial counts were found in the bladder (P ... administered singly, did not have any effect in the UTI model. Apparently, the antibacterial effect of the organic acids from cranberry juice on UTI can be obtained by administering a combination of malic acid and either citric or quinic acid. This study show for the first time that cranberry juice reduce E...

  20. A comparative study of clinical manifestations, haematological and serological responses after experimental infection with Anaplasma phagocytophilum in two Norwegian sheep breeds

    Directory of Open Access Journals (Sweden)

    Sandstedt Karin

    2011-02-01

    Full Text Available Abstract Background It has been questioned if the old native Norwegian sheep breed, Old Norse Sheep (also called Norwegian Feral Sheep, normally distributed on coastal areas where ticks are abundant, is more protected against tick-borne infections than other Norwegian breeds due to a continuously high selection pressure on pasture. The aim of the present study was to test this hypothesis in an experimental infection study. Methods Five-months-old lambs of two Norwegian sheep breeds, Norwegian White (NW sheep and Old Norse (ON sheep, were experimentally infected with a 16S rRNA genetic variant of Anaplasma phagocytophilum (similar to GenBank accession number M73220. The experiment was repeated for two subsequent years, 2008 and 2009, with the use of 16 lambs of each breed annually. Ten lambs of each breed were inoculated intravenously each year with 0.4 ml A. phagocytophilum-infected blood containing approximately 0.5 × 106 infected neutrophils/ml. Six lambs of each breed were used as uninfected controls. Half of the primary inoculated lambs in each breed were re-challenged with the same infectious dose at nine (2008 and twelve (2009 weeks after the first challenge. The clinical, haematological and serological responses to A. phagocytophilum infection were compared in the two sheep breeds. Results The present study indicates a difference in fever response and infection rate between breeds of Norwegian sheep after experimental infection with A. phagocytophilum. Conclusion Although clinical response seems to be less in ON-lambs compared to NW-lambs, further studies including more animals are needed to evaluate if the ON-breed is more protected against tick-borne infections than other Norwegian breeds.

  1. The subunit structure of the extracellular hemoglobin of Biomphalaria glabrata

    International Nuclear Information System (INIS)

    Arndt, Marcio H.L.; Naves, Cristiani F.; Xavier, Luciana P.; Santoro, Marcelo M.

    1997-01-01

    Full text. The hemoglobin of Biomphalaria glabrata was purified to homogeneity by a two step purification protocol using a gel filtration column (Superose 6 HR/Pharmacia ) followed by an anion exchange chromatography (MONO-Q Sepharose/Pharmacia). The dissociation products were analysed by a 5 - 15 % Polyacrylamide gel electrophoresis containing Sodium Dodecyl Sulfate (SDS-PAGE) giving a band of 270 K Daltons and a band of 180 K Daltons after reduction with β-mercaptoethanol. The same profile was obtained in a 3.5 % Agarose gel electrophoresis containing SDS (SDS-AGE) showing additional bands of higher molecular weight. These bands were proposed to be monomers, dimers and trimers and, after reduction in a Bidimensional SDS-AGE, the proposed monomers and dimers were decomposed in two and four bands that were interpreted as 1 - 4 chains. The hemoglobin was digested by four different proteases ( Thrombin, Trypsin, Chymotrypsin and Subtilisin ) showing several equivalent fragments with molecular weights multiples of its minimum molecular weight ( 17.7 K Daltons). The circular dichroism spectrum of the protein showed a characteristic high α-helix content. We proposed that this hemoglobin is a pentamer of approx. 360 K Daltons subunits each formed by two 180 K Daltons chains linked in pairs by disulfide bridges and each of these chains comprises ten Heme binding domains. These data were compared to other Planorbidae extracellular hemoglobins. Up to now, the quaternary structure of this hemoglobin (shape and disposition of the subunits) is unknown. It is intended to elucidate its structure by Small Angle X-Ray Scattering in Brazilian National Laboratory of Synchrotron Light (LNLS). (author)

  2. Experimental Infection and Detection of Necrotizing Hepatopancreatitis Bacterium in the American Lobster Homarus americanus

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    Luz A. Avila-Villa

    2012-01-01

    Full Text Available Necrotizing hepatopancreatitis bacterium (NHPB is an obligated intracellular bacteria causing severe hepatopancreatic damages and mass mortalities in penaeid shrimp. The worldwide distribution of penaeid shrimp as alien species threatens the life cycle of other crustacean species. The aim of the experiment was to evaluate the possibility of experimentally infecting the American lobster (Homarus americanus with NHPB extracted from shrimp hepatopancreas. Homogenates from infected shrimp were fed by force to lobsters. Other group of lobsters was fed with homogenates of NHPB-free hepatopancreas. After the 15th day from initial inoculation, the presence of NHPB was detected by polymerase chain reaction in feces and hepatopancreas from lobsters inoculated with infected homogenates. Necrotized spots were observed in the surface of lobster hepatopancreas. In contrast, lobsters fed on NHPB-free homogenates resulted negative for NHPB. Evidence suggests the plasticity of NHPB which can infect crustacean from different species and inhabiting diverse latitudes. Considering the results, the American lobster could be a good candidate to maintain available NHPB in vivo.

  3. Experimental Infection and Detection of Necrotizing Hepatopancreatitis Bacterium in the American Lobster Homarus americanus

    Science.gov (United States)

    Avila-Villa, Luz A.; Gollas-Galván, Teresa; Martínez-Porchas, Marcel; Mendoza-Cano, Fernando; Hernández-López, Jorge

    2012-01-01

    Necrotizing hepatopancreatitis bacterium (NHPB) is an obligated intracellular bacteria causing severe hepatopancreatic damages and mass mortalities in penaeid shrimp. The worldwide distribution of penaeid shrimp as alien species threatens the life cycle of other crustacean species. The aim of the experiment was to evaluate the possibility of experimentally infecting the American lobster (Homarus americanus) with NHPB extracted from shrimp hepatopancreas. Homogenates from infected shrimp were fed by force to lobsters. Other group of lobsters was fed with homogenates of NHPB-free hepatopancreas. After the 15th day from initial inoculation, the presence of NHPB was detected by polymerase chain reaction in feces and hepatopancreas from lobsters inoculated with infected homogenates. Necrotized spots were observed in the surface of lobster hepatopancreas. In contrast, lobsters fed on NHPB-free homogenates resulted negative for NHPB. Evidence suggests the plasticity of NHPB which can infect crustacean from different species and inhabiting diverse latitudes. Considering the results, the American lobster could be a good candidate to maintain available NHPB in vivo. PMID:22645497

  4. Controlled chaos of polymorphic mucins in a metazoan parasite (Schistosoma mansoni interacting with its invertebrate host (Biomphalaria glabrata.

    Directory of Open Access Journals (Sweden)

    Emmanuel Roger

    Full Text Available Invertebrates were long thought to possess only a simple, effective and hence non-adaptive defence system against microbial and parasitic attacks. However, recent studies have shown that invertebrate immunity also relies on immune receptors that diversify (e.g. in echinoderms, insects and mollusks (Biomphalaria glabrata. Apparently, individual or population-based polymorphism-generating mechanisms exists that permit the survival of invertebrate species exposed to parasites. Consequently, the generally accepted arms race hypothesis predicts that molecular diversity and polymorphism also exist in parasites of invertebrates. We investigated the diversity and polymorphism of parasite molecules (Schistosoma mansoni Polymorphic Mucins, SmPoMucs that are key factors for the compatibility of schistosomes interacting with their host, the mollusc Biomphalaria glabrata. We have elucidated the complex cascade of mechanisms acting both at the genomic level and during expression that confer polymorphism to SmPoMuc. We show that SmPoMuc is coded by a multi-gene family whose members frequently recombine. We show that these genes are transcribed in an individual-specific manner, and that for each gene, multiple splice variants exist. Finally, we reveal the impact of this polymorphism on the SmPoMuc glycosylation status. Our data support the view that S. mansoni has evolved a complex hierarchical system that efficiently generates a high degree of polymorphism-a "controlled chaos"-based on a relatively low number of genes. This contrasts with protozoan parasites that generate antigenic variation from large sets of genes such as Trypanosoma cruzi, Trypanosoma brucei and Plasmodium falciparum. Our data support the view that the interaction between parasites and their invertebrate hosts are far more complex than previously thought. While most studies in this matter have focused on invertebrate host diversification, we clearly show that diversifying mechanisms also

  5. Controlled Chaos of Polymorphic Mucins in a Metazoan Parasite (Schistosoma mansoni) Interacting with Its Invertebrate Host (Biomphalaria glabrata)

    Science.gov (United States)

    Roger, Emmanuel; Grunau, Christoph; Pierce, Raymond J.; Hirai, Hirohisa; Gourbal, Benjamin; Galinier, Richard; Emans, Rémi; Cesari, Italo M.; Cosseau, Céline; Mitta, Guillaume

    2008-01-01

    Invertebrates were long thought to possess only a simple, effective and hence non-adaptive defence system against microbial and parasitic attacks. However, recent studies have shown that invertebrate immunity also relies on immune receptors that diversify (e.g. in echinoderms, insects and mollusks (Biomphalaria glabrata)). Apparently, individual or population-based polymorphism-generating mechanisms exists that permit the survival of invertebrate species exposed to parasites. Consequently, the generally accepted arms race hypothesis predicts that molecular diversity and polymorphism also exist in parasites of invertebrates. We investigated the diversity and polymorphism of parasite molecules (Schistosoma mansoni Polymorphic Mucins, SmPoMucs) that are key factors for the compatibility of schistosomes interacting with their host, the mollusc Biomphalaria glabrata. We have elucidated the complex cascade of mechanisms acting both at the genomic level and during expression that confer polymorphism to SmPoMuc. We show that SmPoMuc is coded by a multi-gene family whose members frequently recombine. We show that these genes are transcribed in an individual-specific manner, and that for each gene, multiple splice variants exist. Finally, we reveal the impact of this polymorphism on the SmPoMuc glycosylation status. Our data support the view that S. mansoni has evolved a complex hierarchical system that efficiently generates a high degree of polymorphism—a “controlled chaos”—based on a relatively low number of genes. This contrasts with protozoan parasites that generate antigenic variation from large sets of genes such as Trypanosoma cruzi, Trypanosoma brucei and Plasmodium falciparum. Our data support the view that the interaction between parasites and their invertebrate hosts are far more complex than previously thought. While most studies in this matter have focused on invertebrate host diversification, we clearly show that diversifying mechanisms also exist on

  6. Fasciola hepatica: comparative metacercarial productions in experimentally-infected Galba truncatula and Pseudosuccinea columella

    Directory of Open Access Journals (Sweden)

    Vignoles Philippe

    2015-01-01

    Full Text Available As large numbers of metacercariae of Fasciola hepatica are necessary for research, experimental infections of Galba truncatula and Pseudosuccinea columella with this digenean were carried out to determine the better intermediate host for metacercarial production and, consequently, the most profitable snail for decreasing the cost price of these larvae. Pre-adult snails (4 mm in shell height originating from two populations per lymnaeid species were individually exposed to two or five miracidia, raised at 23 °C and followed for cercarial shedding up to their death. Compared to values noted in G. truncatula, the survival of P. columella on day 30 post-exposure was significantly greater, while the prevalence of F. hepatica infection was significantly lower. In the four P. columella groups, metacercarial production was significantly greater than that noted in the four groups of G. truncatula (347–453 per cercariae-shedding snail versus 163–275, respectively. Apart from one population of G. truncatula, the use of five miracidia per snail at exposure significantly increased the prevalence of F. hepatica in P. columella and the other population of G. truncatula, whereas it did not have any clear effect on the mean number of metacercariae. The use of P. columella for experimental infections with F. hepatica resulted in significantly higher metacercarial production than that noted with G. truncatula, in spite of a lower prevalence for the former lymnaeid. This finding allows for a significant decrease in the cost price of these larvae for commercial production.

  7. Candida glabrata binds to glycosylated and lectinic receptors on the coronary endothelial luminal membrane and inhibits flow sense and cardiac responses to agonists.

    Science.gov (United States)

    Torres-Tirado, David; Knabb, Maureen; Castaño, Irene; Patrón-Soberano, Araceli; De Las Peñas, Alejandro; Rubio, Rafael

    2016-01-01

    Candida glabrata (CG) is an opportunistic fungal pathogen that initiates infection by binding to host cells via specific lectin-like adhesin proteins. We have previously shown the importance of lectin-oligosaccharide binding in cardiac responses to flow and agonists. Because of the lectinic-oligosaccharide nature of CG binding, we tested the ability of CG to alter the agonist- and flow-induced changes in cardiac function in isolated perfused guinea pig hearts. Both transmission and scanning electron microscopy showed strong attachment of CG to the coronary endothelium, even after extensive washing. CG shifted the coronary flow vs. auricular-ventricular (AV) delay relationship upward, indicating that greater flow was required to achieve the same AV delay. This effect was completely reversed with mannose, partially reversed with galactose and N-acetylgalactosamine, but hyaluronan had no effect. Western blot analysis was used to determine binding of CG to isolated coronary endothelial luminal membrane (CELM) receptors, and the results indicate that flow-sensitive CELM receptors, ANG II type I, α-adrenergic 1A receptor, endothelin-2, and VCAM-1 bind to CG. In addition, CG inhibited agonist-induced effects of bradykinin, angiotensin, and phenylephrine on AV delay, coronary perfusion pressure, and left ventricular pressure. Mannose reversed the inhibitory effects of CG on the agonist responses. These results suggest that CG directly binds to flow-sensitive CELM receptors via lectinic-oligosaccharide interactions with mannose and disrupts the lectin-oligosaccharide binding necessary for flow-induced cardiac responses. Copyright © 2016 the American Physiological Society.

  8. Analysis of experimental mink enteritis virus infection in mink: in situ hybridization, serology, and histopathology

    DEFF Research Database (Denmark)

    Uttenthal, Åse; Larsen, S; Lund, E

    1990-01-01

    Strand-specific hybridization probes were used in in situ hybridization studies to localize cells containing mink enteritis virus (MEV) virion DNA or MEV replicative-form DNA and mRNA. Following the experimental MEV infection of 3-month-old unvaccinated mink, a significant increase in serum antib...

  9. Pathogenesis of canine distemper virus in experimentally infected raccoon dogs, foxes, and minks.

    Science.gov (United States)

    Zhao, Jianjun; Shi, Ning; Sun, Yangang; Martella, Vito; Nikolin, Veljko; Zhu, Chunsheng; Zhang, Hailing; Hu, Bo; Bai, Xue; Yan, Xijun

    2015-10-01

    Canine distemper virus (CDV) infects a broad range of carnivores and causes a highly contagious disease with severe immunosuppression. The disease severity markedly varies in different species. To investigate the pathogenesis of CDV in raccoon dog (Nyctereutes procyonoides), fox (Vulpes vulpes) and mink (Neovison vison) species, three groups of CDV sero-negative animals were infected with CDV strain LN(10)1. This CDV strain belongs to the Asia-1 genotype, which is epidemiologically predominant in carnivores in China. CDV infection provoked marked differences in virulence in the three species that were studied. Raccoon dogs developed fever, severe conjunctivitis, and pathological lesions, with 100% (5/5) mortality and with high viral RNA loads in organs within 15 days post infection (dpi). In infected foxes, the onset of the disease was delayed, with 40% (2/5) mortality by 21 dpi. Infected minks developed only mild clinical signs and pathological lesions, and mortality was not observed. Raccoon dogs and foxes showed more severe immune suppression (lymphopenia, decreased lymphocyte proliferation, viremia and low-level virus neutralizing antibodies) than minks. We also observed a distinct pattern of cytokine mRNA transcripts at different times after infection. Decreased IFN-γ and IL-4 mRNA responses were evident in the animals with fatal disease, while up-regulation of these cytokines was observed in the animals surviving the infection. Increased TNF-α response was detected in animals with mild or severe clinical signs. Based on the results, we could distinguish three different patterns of disease after experimental CDV infection, e.g. a mild form in minks, a moderate form in foxes and a severe disease in raccoon dogs. The observed differences in susceptibility to CDV could be related to distinct host cytokine profiles. Comparative evaluation of CDV pathogenesis in various animal species is pivotal to generate models suitable for the evaluation of CDV

  10. Detection and quantification of pestivirus in experimentally infected pregnant ewes and their progeny

    Directory of Open Access Journals (Sweden)

    Juste Ramón A

    2009-11-01

    Full Text Available Abstract Background Border disease virus (BDV causes important reproductive losses, and eradication strategies focus on the identification and removal of persistently infected animals arising after in uterine infection. BDV infection dynamics were studied in 13 ewes experimentally infected with BDV-4 genotype at 3 phases of pregnancy [days 108 (group A, 76 (group B and 55 (group C] by quantification of viral RNA in blood collected on days -1 to parturition using quantitative real-time RT-PCR (qRT-PCR. Viral RNA loads were also measured in blood/foetal fluid and tissue samples from their offspring at lambing (3 foetuses, 7 stillborns, 15 lambs. qRT-PCR results were compared with those obtained by conventional RT-PCR and used to predict persistent infections. Results Viral RNA was detected in the ewes between days 2-15 p.i. The viraemia reached its highest peak between days 6-7 p.i. with a second peak at days 11-12 p.i. qRT-PCR was significantly faster to perform (less than 1 h than conventional RT-PCR and detected BDV RNA in more ewes, being detection more continuous and prolonged in time. The virus was detected in peripheral blood in a higher percentage of lambs than in tissues, where differences in viral genome copies were more marked. Skin and cerebral cortex showed the highest viral RNA loads, and spleen and spinal cord the lowest. High viral RNA loads were observed in several animals in group B and all in group C, infected during middle and early foetal development, respectively, but also in one lamb from group A, infected during late foetal development. Serology and viral genome copy number estimates in blood and tissues were used to establish a quantitative cut-off threshold for transient viraemia. Conclusion Viral RNA quantification showed potential for the discrimination between persistent infections and transient viraemia using single-time point blood sampling and raised questions regarding foetal immune system development and the

  11. Histological assessment of granulomas in natural and experimental Schistosoma mansoni infections using whole slide imaging.

    Directory of Open Access Journals (Sweden)

    Kátia B Amaral

    Full Text Available The pathology of schistosomiasis mansoni, a neglected tropical disease of great clinical and socioeconomic importance, results from the parasite eggs that become trapped in host tissues, particularly in the liver and intestines. Continuous antigenic stimulation from these eggs leads to recruitment of inflammatory cells to the sites of infection with formation of periovular granulomas. These complex structures have variable size and composition and are the most striking histopathological feature of schistosomiasis mansoni. However, evaluation of granulomas by conventional microscopy methods is time-consuming and limited, especially in large-scale studies. Here, we used high resolution Whole Slide Imaging (WSI, which allows fast scanning of entire histological slides, and multiple morphometric evaluations, to assess the granulomatous response elicited in target organs (liver, small and large intestines of two models of schistosomiasis mansoni. One of the advantages of WSI, also termed virtual microscopy, is that it generates images that simultaneously offer high resolution and a wide field of observation. By using a model of natural (Nectomys squamipes, a wild reservoir captured from endemic areas in Brazil and experimental (Swiss mouse infection with Schistosoma mansoni, we provided the first detailed WSI characterization of granulomas and other pathological aspects. WSI and quantitative analyses enabled a fast and reliable assessment of the number, evolutional types, frequency and areas of granulomas and inflammatory infiltrates and revealed that target organs are differentially impacted by inflammatory responses in the natural and experimental infections. Remarkably, high-resolution analysis of individual eosinophils, key cells elicited by this helminthic infection, showed a great difference in eosinophil numbers between the two infections. Moreover, features such as the intestinal egg path and confluent granulomas were uncovered. Thus, WSI may

  12. Effect of wide spectrum anti-helminthic drugs upon Schistosoma mansoni experimentally infected mice

    Directory of Open Access Journals (Sweden)

    PANCERA Christiane Finardi

    1997-01-01

    Full Text Available Mebendazole, albendazole, levamisole and thiabendazole are well known as active drugs against several nematode species, and against cestodes as well, when the first two drugs are considered. None of the drugs have proven activity, however, against trematodes. We tested the effect of these drugs on the fecal shedding of schistosome eggs and the recovering of adult schistosomes, after portal perfusion in Schistosoma mansoni experimentally infected mice. Balb/c mice infected with 80 S. mansoni cercariae were divided into three groups, each in turn subdivided into four other groups, for each tested drug. The first group was treated with each one of the studied drugs 25 days after S. mansoni infection; the second group was submitted to treatment with each one of the drugs 60 days after infection. Finally, the third group, considered as control, received no treatment. No effect upon fecal shedding of S. mansoni eggs and recovering of schistosomes after portal perfusion was observed when mice were treated with either mebendazole or albendazole. Mice treated with either levamisole or thiabendazole, on the other hand, showed a significant reduction in the recovering of adult schistosomes after portal perfusion, mainly when both drugs were given during the schistosomula evolution period, i.e., 25 days after cercariae penetration, probably due to unspecific immunomodulation

  13. Experimental reproduction of beak atrophy and dwarfism syndrome by infection in cherry valley ducklings with a novel goose parvovirus-related parvovirus.

    Science.gov (United States)

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

    2016-02-01

    Infection of clinically susceptible ducks, including cherry valley and Muscovy ducks, with a novel goose parvovirus (GPV)-related virus (N-GPV) can result in beak atrophy and dwarfism syndrome (BADS). To obtain new insights into the host range and pathogenic potential of this novel waterfowl parvovirus, cherry valley ducklings (n=20) were experimentally infected with N-GPV strain SDLC01. An equal number of ducklings served as uninfected controls. The appearance of clinical signs, histopathological changes, viral shedding, and seroconversion was monitored for 20 days post-infection. Infection status of all ducks was monitored using indirect ELISA, virus neutralization test, nested PCR, clinical indicators, and microscopic examination. Three ducks developed the typical clinical, gross, and histological changes of BADS. By study day 6, the infected ducks had seroconverted to N-GPV. The antibodies raised were neutralizing against the SDLC01 strain in vitro. Here we successfully developed an experimental infection model for studying the pathogenicity and role of N-GPV in BADS. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Therapy of the experimental infection by Strongyloides venezuelensis in rats with injectable ivermectin or levamizole

    Directory of Open Access Journals (Sweden)

    Rubens Campos

    1989-02-01

    Full Text Available For the therapy of human strongyloidiasis, are necessary effective drugs to eliminate both larvae and adult worm parasitism, which may also be used by parenteral route, to obviate the particular conditions presented by many patients. A study based on the experimental infection by Strongyloides venezuelensis in rats was done, administering injectable ivermectin or levamizole. Both drugs were shown to be active, when used in single doses of 0.2 to 0.5 mg/kg of ivermectin, or 26 mg/kg for levamizole. Ivermectin was slightly more effective as far as larval stage of the infection is concerned, and the same happened for levamisole for the adult worm stage. Promising perspectives are visualized to improve the therapy of patients with serious disseminated infection by Strongyloides stercoralis.

  15. Novel experimental Pseudomonas aeruginosa lung infection model mimicking long-term host-pathogen interactions in cystic fibrosis

    DEFF Research Database (Denmark)

    Moser, Claus; van Gennip, Maria; Bjarnsholt, Thomas

    2009-01-01

    Moser C, van Gennip M, Bjarnsholt T, Jensen PO, Lee B, Hougen HP, Calum H, Ciofu O, Givskov M, Molin S, Hoiby N. Novel experimental Pseudomonas aeruginosa lung infection model mimicking long-term host-pathogen interactions in cystic fibrosis. APMIS 2009; 117: 95-107. The dominant cause of premature...... death in patients suffering from cystic fibrosis (CF) is chronic lung infection with Pseudomonas aeruginosa. The chronic lung infection often lasts for decades with just one clone. However, as a result of inflammation, antibiotic treatment and different niches in the lungs, the clone undergoes...... and 2003) of the chronic lung infection of one CF patient using the seaweed alginate embedment model. The results showed that the non-mucoid clones reduced their virulence over time, resulting in faster clearing of the bacteria from the lungs, improved pathology and reduced pulmonary production...

  16. The epidemiology and outcomes of invasive Candida infections among organ transplant recipients in the United States: results of the Transplant-Associated Infection Surveillance Network (TRANSNET).

    Science.gov (United States)

    Andes, David R; Safdar, Nasia; Baddley, John W; Alexander, Barbara; Brumble, Lisa; Freifeld, Allison; Hadley, Susan; Herwaldt, Loreen; Kauffman, Carol; Lyon, G Marshall; Morrison, Vicki; Patterson, Thomas; Perl, Trish; Walker, Randall; Hess, Tim; Chiller, Tom; Pappas, Peter G

    2016-12-01

    Invasive candidiasis (IC) is a common cause of mortality in solid organ transplant recipients (OTRs), but knowledge of epidemiology in this population is limited. The present analysis describes data from 15 US centers that prospectively identified IC from nearly 17 000 OTRs. Analyses were undertaken to determine predictors of infection and mortality. A total of 639 cases of IC were identified. The most common species was Candida albicans (46.3%), followed by Candida glabrata (24.4%) and Candida parapsilosis (8.1%). In 68 cases >1 species was identified. The most common infection site was bloodstream (44%), followed by intra-abdominal (14%). The most frequently affected allograft groups were liver (41.1%) and kidney (35.3%). All-cause mortality at 90 days was 26.5% for all species and was highest for Candida tropicalis (44%) and C. parapsilosis (35.2%). Non-white race and female gender were more commonly associated with non-albicans species. A high rate of breakthrough IC was seen in patients receiving antifungal prophylaxis (39%). Factors associated with mortality include organ dysfunction, lung transplant, and treatment with a polyene antifungal. The only modifiable factor identified was choice of antifungal drug class based upon infecting Candida species. These data highlight the common and distinct features of IC in OTRs. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  17. Experimental infection of plants with an herbivore-associated bacterial endosymbiont influences herbivore host selection behavior.

    Directory of Open Access Journals (Sweden)

    Thomas Seth Davis

    Full Text Available Although bacterial endosymbioses are common among phloeophagous herbivores, little is known regarding the effects of symbionts on herbivore host selection and population dynamics. We tested the hypothesis that plant selection and reproductive performance by a phloem-feeding herbivore (potato psyllid, Bactericera cockerelli is mediated by infection of plants with a bacterial endosymbiont. We controlled for the effects of herbivory and endosymbiont infection by exposing potato plants (Solanum tuberosum to psyllids infected with "Candidatus Liberibacter solanacearum" or to uninfected psyllids. We used these treatments as a basis to experimentally test plant volatile emissions, herbivore settling and oviposition preferences, and herbivore population growth. Three important findings emerged: (1 plant volatile profiles differed with respect to both herbivory and herbivory plus endosymbiont infection when compared to undamaged control plants; (2 herbivores initially settled on plants exposed to endosymbiont-infected psyllids but later defected and oviposited primarily on plants exposed only to uninfected psyllids; and (3 plant infection status had little effect on herbivore reproduction, though plant flowering was associated with a 39% reduction in herbivore density on average. Our experiments support the hypothesis that plant infection with endosymbionts alters plant volatile profiles, and infected plants initially recruited herbivores but later repelled them. Also, our findings suggest that the endosymbiont may not place negative selection pressure on its host herbivore in this system, but plant flowering phenology appears correlated with psyllid population performance.

  18. Assessment of experimental infection for dogs usingGallus gallus chorioallantoic membranes inoculated withNeospora caninum

    Directory of Open Access Journals (Sweden)

    Alexandre Dias Munhoz

    Full Text Available The aim of this study was to evaluate parasitism kinetics and tissue lesions in the first week of infection by Neospora caninum in dogs fed Gallus gallus chorioallantoic membranes (CMs previously infected in ovo. Five two-month-old pups were used. Each dog was given five CMs that were previously infected with N. caninum via the oral route. Four animals were euthanized in the first week of infection. All four dogs had their stools examined one week prior to and up to the day they were euthanized. The stools of the uneuthanized dog were collected for 30 days. After euthanasia, organ sections were utilized for histopathology, immunohistochemistry, indirect immunofluorescent tissue reactions, PCR and real-time PCR to detect parasites. Necropsy revealed that the small and large intestines, spleen, and lungs were affected. No oocysts orN. caninum DNA were identified in the stool samples. Real-time PCR was the most sensitive technique used to detect the protozoa in tissues, which were identified in 41% of the analyzed samples. Our results indicate that an experimental model using previously infected CMs appears to be a useful model for the study of the host-parasite relationship during the infection's acute phase.

  19. Experimental infection of two South American reservoirs with four distinct strains of Trypanosoma cruzi

    Science.gov (United States)

    Roellig, Dawn M.; McMillan, Katherine; Ellis, Angela E.; Vandeberg, John L.; Champagne, Donald E.; Yabsley, Michael J.

    2010-01-01

    SUMMARY Trypanosoma cruzi (Tc), the causative agent of Chagas disease, is a diverse species with 2 primary genotypes, TcI and TcII, with TcII further subdivided into 5 subtypes (IIa–e). This study evaluated infection dynamics of 4 genetically and geographically diverse T. cruzi strains in 2 South American reservoirs, degus (Octodon degus) and grey short-tailed opossums (Monodelphis domestica). Based on prior suggestions of a genotype-host association, we hypothesized that degus (placental) would more readily become infected with TcII strains while short-tailed opossums (marsupial) would be a more competent reservoir for a TcI strain. Individuals (n = 3) of each species were intraperitoneally inoculated with T. cruzi trypomastigotes of TcIIa [North America (NA)-raccoon (Procyon lotor) origin], TcI [NA-Virginia opossum (Didelphis virginiana)], TcIIb [South America (SA)-human], TcIIe (SA-Triatoma infestans), or both TcI and TcIIa. Parasitaemias in experimentally infected degus peaked earlier (7–14 days post-inoculation (p.i.)) compared with short-tailed opossums (21–84 days p.i.). Additionally, peak parasitaemias were higher in degus; however, the duration of detectable parasitaemias for all strains, except TcIIa, was greater in short-tailed opossums. Infections established in both host species with all genotypes, except for TcIIa, which did not establish a detectable infection in short-tailed opossums. These results indicate that both South American reservoirs support infections with these isolates from North and South America; however, infection dynamics differed with host and parasite strain. PMID:20128943

  20. Comparative Experimental Infection Study in Dogs with Ehrlichia canis, E. chaffeensis, Anaplasma platys and A. phagocytophilum.

    Science.gov (United States)

    Nair, Arathy D S; Cheng, Chuanmin; Ganta, Chanran K; Sanderson, Michael W; Alleman, Arthur R; Munderloh, Ulrike G; Ganta, Roman R

    2016-01-01

    Dogs acquire infections with the Anaplasmataceae family pathogens, E. canis, E. chaffeensis, E. ewingii, A. platys and A. phagocytophilum mostly during summer months when ticks are actively feeding on animals. These pathogens are also identified as causing diseases in people. Despite the long history of tick-borne diseases in dogs, much remains to be defined pertaining to the clinical and pathological outcomes of infections with these pathogens. In the current study, we performed experimental infections in dogs with E. canis, E. chaffeensis, A. platys and A. phagocytophilum. Animals were monitored for 42 days to evaluate infection-specific clinical, hematological and pathological differences. All four pathogens caused systemic persistent infections detectible throughout the 6 weeks of infection assessment. Fever was frequently detected in animals infected with E. canis, E. chaffeensis, and A. platys, but not in dogs infected with A. phagocytophilum. Hematological differences were evident in all four infected groups, although significant overlap existed between the groups. A marked reduction in packed cell volume that correlated with reduced erythrocytes and hemoglobin was observed only in E. canis infected animals. A decline in platelet numbers was common with E. canis, A. platys and A. phagocytophilum infections. Histopathological lesions in lung, liver and spleen were observed in all four groups of infected dogs; infection with E. canis had the highest pathological scores, followed by E. chaffeensis, then A. platys and A. phagocytophilum. All four pathogens induced IgG responses starting on day 7 post infection, which was predominantly comprised of IgG2 subclass antibodies. This is the first detailed investigation comparing the infection progression and host responses in dogs after inoculation with four pathogens belonging to the Anaplasmataceae family. The study revealed a significant overlap in clinical, hematological and pathological changes resulting from the

  1. Evaluation of metaphylactic RNA interference to prevent equine herpesvirus type 1 infection in experimental herpesvirus myeloencephalopathy in horses.

    Science.gov (United States)

    Perkins, Gillian A; Van de Walle, Gerlinde R; Pusterla, Nicola; Erb, Hollis N; Osterrieder, Nikolaus

    2013-02-01

    To evaluate metaphylactic RNA interference to prevent equine herpesvirus type 1 (EHV-1) infection in experimental herpesvirus myeloencephalopathy in horses and to determine whether horses infected with a neuropathogenic strain of the virus that develop equine herpesvirus myeloencephalopathy (EHM) have differences in viremia. 13 seronegative horses. EHV-1 strain Ab4 was administered intranasally on day 0, and small interfering RNAs (siRNAs [EHV-1 specific siRNAs {n = 7} or an irrelevant siRNA {6}]) were administered intranasally 24 hours before and 12, 24, 36, and 48 hours after infection. Physical and neurologic examinations, nasal swab specimens, and blood samples were collected for virus isolation and quantitative PCR assay. Data from the study were combined with data from a previous study of 14 horses. No significant difference was detected in clinical variables, viremia, or detection of EHV-1 in nasal swab specimens of horses treated with the EHV-1 targeted siRNAs (sigB3-siOri2) versus controls. No significant differences in viremia were detected between horses that developed EHM and those that did not. Administration of siRNAs targeted against EHV-1 around the time of EHV-1 infection was not protective with this experimental design. Horses infected with the neuropathogenic EHV-1 strain Ab4 that developed EHM did not have a more pronounced viremia.

  2. Use of quantitative real-time RT-PCR to investigate the correlation between viremia and viral shedding of canine distemper virus, and infection outcomes in experimentally infected dogs.

    Science.gov (United States)

    Sehata, Go; Sato, Hiroaki; Ito, Toshihiro; Imaizumi, Yoshitaka; Noro, Taichi; Oishi, Eiji

    2015-07-01

    We used real-time RT-PCR and virus titration to examine canine distemper virus (CDV) kinetics in peripheral blood and rectal and nasal secretions from 12 experimentally infected dogs. Real-time RT-PCR proved extremely sensitive, and the correlation between the two methods for rectal and nasal (r=0.78, 0.80) samples on the peak day of viral RNA was good. Although the dogs showed diverse symptoms, viral RNA kinetics were similar; the peak of viral RNA in the symptomatic dogs was consistent with the onset of symptoms. These results indicate that real-time RT-PCR is sufficiently sensitive to monitor CDV replication in experimentally infected dogs regardless of the degree of clinical manifestation and suggest that the peak of viral RNA reflects active CDV replication.

  3. Effects of Ichthyophonus hoferi on condition indices and blood chemistry of experimentally infected rainbow trout (Oncorhynchus mykiss).

    Science.gov (United States)

    Rand, T G; Cone, D K

    1990-07-01

    Body condition, hepatosomatic index and blood chemistry of Oncorhynchus mykiss experimentally infected with a tissue dwelling fish pathogenic fungus, Ichthyophonus hoferi, were monitored over a 6 wk period. This was to determine whether the infection constituted a stress manifest by changes in the hypothalamic-pituitary interrenal axis, and especially plasma cortisol levels. Infection caused anaemia and leucopenia but did not change the condition, hepatosomatic indices, or plasma chloride, cholesterol, cortisol, creatinine, glucose, osmolarity, potassium, total protein, sodium and T4. It is suggested that increased cortisol levels may not be a normal component of the stress response of fish to disease caused by invasive infectious agents.

  4. Oral Yeast Colonization and Fungal Infections in Peritoneal Dialysis Patients: A Pilot Study

    Directory of Open Access Journals (Sweden)

    Liliana Simões-Silva

    2017-01-01

    Full Text Available Peritonitis and exit-site infections are important complications in peritoneal dialysis (PD patients that are occasionally caused by opportunistic fungi inhabiting distant body sites. In this study, the oral yeast colonization of PD patients and the antifungal susceptibility profile of the isolated yeasts were accessed and correlated with fungal infection episodes in the following 4 years. Saliva yeast colonization was accessed in 21 PD patients and 27 healthy controls by growth in CHROMagar-Candida® and 18S rRNA/ITS sequencing. PD patients presented a lower oral yeast prevalence when compared to controls, namely, Candida albicans. Other species were also isolated, Candida glabrata and Candida carpophila. The antifungal susceptibility profiles of these isolates revealed resistance to itraconazole, variable susceptibility to caspofungin, and higher MIC values of posaconazole compared to previous reports. The 4-year longitudinal evaluation of these patients revealed Candida parapsilosis and Candida zeylanoides as PD-related exit-site infectious agents, but no correlation was found with oral yeast colonization. This pilot study suggests that oral yeast colonization may represent a limited risk for fungal infection development in PD patients. Oral yeast isolates presented a variable antifungal susceptibility profile, which may suggest resistance to some second-line drugs, highlighting the importance of antifungal susceptibility assessment in the clinical practice.

  5. Effects of single or trickle Haemonchus contortus experimental infection on digestibility and host responses of naïve Creole kids reared indoor.

    Science.gov (United States)

    Bambou, J C; Cei, W; Camous, S; Archimède, H; Decherf, A; Philibert, L; Barbier, C; Mandonnet, N; González-García, E

    2013-01-31

    The objective of this study was to compare the effects of the type of Haemonchus contortus experimental infection (trickle infection, TI versus single infection, SI) on feed intake, nutrients digestibility, parasitological and haematological measures, and plasma leptin in Creole kids. The animals were infected over 2 periods (challenge 1 and challenge 2) of 6 weeks each, corresponding respectively to the primary and the secondary infection. Periods prior infection (1 week each) were considered as controls. The primary infection was realized with 35 Creole kids (18.40±3.76 kg BW) housed in individual boxes and fed a hay-based diet. The secondary infection continued with 29 kids (21.90±3.40 kg BW) from the initial 35. A total of 6 kids and 8 kids were slaughtered for measuring nematode burden at the end of the primary and the secondary infection, respectively. Measurements of nutrients digestibility were made at 0, 3 and 5 weeks post-infection for both challenges. Faecal egg count (FEC), blood eosinophilia and packed cell volume (PCV) were monitored weekly. Feed intake (dry matter intake, DMI) and nutrients digestibility were negatively affected by H. contortus infection only during the primary infection. Plasma leptin changed significantly over time (P=0.0002) but was not affected by the infection type. Effect of infection type was observed only on crude protein digestibility during the primary infection, which was lower in the TI group (P<0.01). The overall level of blood eosinophilia was significantly higher in the TI group (P<0.0001) during both challenges. The overall FEC mean was significantly higher in the SI compared with the TI groups, during both challenges (P<0.02). These results were related to the mean female length significantly higher in the SI group compared with the TI group during challenge 1 (P=0.004), and the number of adult nematode significantly lower in the TI group compared with the SI group during the challenge 2 (P=0.05). The results

  6. Sympatric and allopatric experimental infections of the planorbid snail Gyraulus chinensis with miracidia of Euparyphium albuferensis (Trematoda: Echinostomatidae).

    Science.gov (United States)

    Muñoz-Antoli, C; Marín, A; Trelis, M; Toledo, R; Esteban, J-G

    2010-12-01

    An experimental infection with echinostomatid miracidia in sympatric or 'local' vs. allopatric or 'away' snail combinations, as a model to examine parasite compatibility, was carried out. We employed Euparyphium albuferensis miracidia to infect Gyraulus chinensis snails, from three different natural parks: Albufera (Valencia, Spain); the Ebro Delta (Tarragona, Spain) and Coto de Doñana (Huelva, Spain). Insignificant differences between the three snail strains were noted for the infection rate and the rhythm of daily cercarial production. However, a significantly higher total cercarial production per snail, patent period and life span were observed in local snails. The different infection characteristics in the three G. chinensis strains considered reveal that E. albuferensis miracidia demonstrate local adaptation.

  7. The effect of gamma radiation and caffeine during the embrionic development of Biomphalaria glabrata

    International Nuclear Information System (INIS)

    Kawano, T.

    1982-01-01

    Some experiments have been carried out, dealing with the interations of caffeine and gamma radiation in the embryonic development of Biomphalaria glabrata. Gamma radiation was used in a dosis of 1000r (exposure time of 1/2 hour) and caffeine concentration was 0.1% (Exposure time of 24 hours). Experiment I. The embryos were irradiated before treatment with caffeine. In this experiment we have observed a synergistic effect between caffeine and gamma radiation. Experiment II. The embryos in the caffeine solution were irradiated 8 hours after the beginning of the experiment. The synergistic effect observed in this experiment was reduced when compared with the first one. Experiment III. In this experiment we made the pre-treatment with caffeine. We have not observed any synergistic effect between the drug and gamma radiation. Experiment IV. Embryos were irradiated, and after 24 hours treated with caffeine during another 24 hours. As in the previous experiment, we have not observed any synergism between the radiation and the drug. According to the above results, we conclude that the synergistic effect between caffeine and gamma radiation in the embryos was more effective when the embryos have been treated with caffeine after irradiation. (Author) [pt

  8. Evaluation of Urinary Tract Infections Due to Candida Species

    Directory of Open Access Journals (Sweden)

    Yeser Karaca Derici

    2016-02-01

    Full Text Available Aim: Although urinary tract infections often caused by bacteria, fungal etiology is detected in a significant number of infections in which Candida is the leading cause. In this study we aimed to evaluate the distribution of Candida strains isolated from urine samples in our hospital. Material and Method: Candida species were identified based on germ tube test, colony morphology on chrom agar Candida (Biomerieux, France and API ID32C AUX (Biomerieux, France commercial kit. Data were analyzed with SPSS 15.0 software for data analysis. Results: During March 2011-March 2014 a total of 109662 urine cultures were evaluated and 24364 samples revealed significant growth. Of the significant growth detected 24364 (22% samples 1096 (4.5% were defined as yeasts. The isolates most frequently detected in this study were C. albicans (50.5%, C. tropicalis (15.9%, C. glabrata (12.7%, C. parapsilosis (7.2%, C. kefyr (5.8%, C. krusei (5.5%. The highest yeast growth was observed in anesthesia intensive care unit. Discussion: In our study, the most frequently isolated species of yeast in the urine was C. albicans. Determination of Candida species and their clinical distributions in hospitals is very important in terms of giving direction to the treatment and measures to be taken.

  9. IL-18 potentiated whole blood IFN-γ assay can identify cell-mediated immune responses towards Lawsonia intracellularis in experimentally infected pigs

    DEFF Research Database (Denmark)

    Riber, Ulla; Jakobsen, Jeanne Toft; Hvass, Henriette Cordes

    Lawsonia intracellularis is an obligate intracellular bacteria causing proliferative enteropathy (PE) in pigs. The infection causes diarrhoea, retarded growth and sudden death in pigs and is one of the most economically important diseases in the swine industry worldwide. The infection is one...... indications that cell-mediated immune responses (CMI) are important for the protection against infections with L. intracellularis and in mice models IFN-γ has been shown to play a key role in the host defence against experimental infections . In L. intracellularis infected pigs, IFN-γ is only sparsely...

  10. Potential Role of Carvedilol in the Cardiac Immune Response Induced by Experimental Infection with Trypanosoma cruzi

    Directory of Open Access Journals (Sweden)

    Aline Luciano Horta

    2017-01-01

    Full Text Available Trypanosoma cruzi causes a cardiac infection characterized by an inflammatory imbalance that could become the inciting factor of the illness. To this end, we evaluated the role of carvedilol, a beta-blocker with potential immunomodulatory properties, on the immune response in C57BL/6 mice infected with VL-10 strain of T. cruzi in the acute phase. Animals (n=40 were grouped: (i not infected, (ii infected, (iii infected + carvedilol, and (iv not infected + carvedilol. We analyzed parameters related to parasitemia, plasma levels of TNF, IL-10, and CCL2, and cardiac histopathology after the administration of carvedilol for 30 days. We did not observe differences in the maximum peaks of parasitemia in the day of their detection among the groups. The plasma TNF was elevated at 60 days of infection in mice treated or not with carvedilol. However, we observed a decreased CCL2 level and increased IL-10 levels in those infected animals treated with carvedilol, which impacted the reduction of the inflammatory infiltration in cardiac tissue. For this experimental model, carvedilol therapy was not able to alter the levels of circulating parasites but modulates the pattern of CCL2 and IL-10 mediators when the VL10 strain of T. cruzi was used in C57BL6 mice.

  11. Experimental infection in Cavia porcellus by infected Amblyomma ovale nymphs with Rickettsia sp. (Atlantic rainforest strain).

    Science.gov (United States)

    Brustolin, Joice Magali; da Silva Krawczak, Felipe; Alves, Marta Elena Machado; Weiller, Maria Amélia; de Souza, Camila Lopes; Rosa, Fábio Brum; Cadore, Gustavo Cauduro; Dos Anjos Lopes, Sônia Terezinha; Labruna, Marcelo Bahia; Vogel, Fernanda Silveira Flores; de Avila Botton, Sônia; Sangioni, Luís Antônio

    2018-03-01

    This study describes experimental infection of guinea pigs (Cavia porcellus) infested with naturally infected Amblyomma ovale nymphs with Rickettsia sp. (Atlantic rainforest strain), and the capacity of A. ovale nymphs to transmit this bacterium. Twenty-six guinea pigs were divided into the following groups: G1, 10 animals infested with uninfected A. ovale nymphs; G2, 10 animals infested with nymphs infected with Rickettsia sp. (Atlantic rainforest strain); and G3, 6 animals without tick infestation. Blood samples were taken 7, 14, 21, and 28 days post-infestation for serological and hematological tests. For histopathological analysis and rickettsial DNA detection, fragments of the spleen, lung, brain, and liver were harvested after euthanasia. The average feeding period for nymphs was 6.6 days for G1 and 6 days for G2. Hemolymph and PCR assays, performed to detect the causative agent in ticks, indicated that in G1, all ticks were negative, and in G2, all nymphs were positive by PCR and 80% (8/10) was positive by hemolymph tests. The only clinical change was skin scarring at the tick attachment site. Hematological parameters indicated leukopenia and total plasma protein (TPP) increased with decreased platelets in G1. In G2, leukocytosis, neutrophilia, monocytosis, an increase in platelets, and reduced TPP were observed. Only G2 guinea pigs were seroconverted (80%; 8/10). Histopathology tests indicated mild, diffuse hemosiderosis and mild, multifocal, follicular hyperplasia in the spleen. Molecular analysis did not detect Rickettsia sp. DNA in C. porcellus tissues. We demonstrated the capacity of A. ovale nymphs to transmit Rickettsia sp. (Atlantic rainforest strain) to guinea pigs.

  12. Immune responsiveness associated with experimental Encephalitozoon intestinalis infection in immunocompetent rats

    Directory of Open Access Journals (Sweden)

    Omalu ICJ

    2007-01-01

    Full Text Available Purpose: Microsporidial infections have been recognized as an increasingly important infection in immuncompromised patients, particularly those infected with HIV/AIDS. This study was designed to study immune responses associated with experimental Encephalitozoon intestinalis infection in immunocompetent rats. Materials and Methods: Thirty-four Rats in 3 groups, A (Control, B (Intraperitoneal and C (Oral were given injections of 0.5 ml of 2 x 10 6 of purified spores of Encephalitotozoon intestinalis spores and were observed for serum specific IgG for 21 days using both direct and indirect ELISA. Results: In indirect ELISA, specific lgG were detected on days 7, 14 and 21 for the group B rats and on day 21 for group C and in direct ELISA method, specific lgG were detected in-group B rats on days 7 and 21, for group C rats on day 21 only, while in the control rats, specific lgG were not detected. There was no significant difference between the direct and indirect methods (df=1, X 2 , P>0.05. E. intestinalis was observed in stool samples of rats in 1/12 (08.33% on days 14 and 21 in group B, and in 4/10 (33.33%, 3/10 (25.00% and 2/10 (16.67% on days 7, 14 and 21 respectively in group C. In group A, which is the control rats, no microsporidia were observed on days 0, 7, 14 and 21. Conclusions: There were no changes in the T-lymphocyte counts of rats prior to and after inoculation with spores. Extensive lesions were observed along the intestinal walls especially on the middle and lower sections of group C rats only.

  13. Antibacterial efficacy of ethyl acetate fraction of Psidium guajava leaf aqueous extract on experimental Escherichia coli (O78) infection in chickens.

    Science.gov (United States)

    Geidam, Y A; Ambali, A G; Onyeyili, P A; Tijjani, M B; Gambo, H I; Gulani, I A

    2015-03-01

    This study was desingned to examine the efficacy of ethyl acetate fraction of aqueous extracted Psidium guajava leaves on chicks experimentally-infected with diarrheagenic strain of Escherichia coli O78. A total of 60 ISA brown male chicks were randomly divided into 6 Groups of ten chicks each in separate cages. Group A was not infected and not treated. Groups B, C and D were infected and treated with extracts at a dose of 25, 50 and 100 mg/kg respectively for 10 days. Group E was infected and treated with oxytetracycline while Group F was infected, but left untreated. Chicks from all groups were closely monitored for clinical signs, body weight change and fecal bacterial shedding load during the course of the experiment. Diarrhea, vents pasted with feces, drop in feed intake accompanied by slow weight gain and decreased activity was observed in infected untreated groups. Groups treated with graded doses of the extract experienced a dose-dependent decreased in severity of the clinical signs shown compared to the infected untreated group. Bacterial shedding load was found to be lower in groups treated with the extract and oxytetracycline than those without intervention. Ethyl acetate soluble fraction of leaf extract of Psidium guajava effectively controlled diarrhea and decreased the severity of other clinical signs caused by experimental E. coli infections in chicks.

  14. In situ hybridization for the detection of infectious laryngotracheitis virus in sections of trachea from experimentally infected chickens

    DEFF Research Database (Denmark)

    Nielsen, O.L.; Handberg, Kurt; Jørgensen, Poul Henrik

    1998-01-01

    An in situ hybridization procedure for the detection of infectious laryngotracheitis virus (ILTV) in experimentally infected chickens is described. Formalin-fixed, paraffin-embedded sections of trachea, taken from chickens on days 3-10 post-inoculation (p.i.) with ILTV were hybridized with a mixt......An in situ hybridization procedure for the detection of infectious laryngotracheitis virus (ILTV) in experimentally infected chickens is described. Formalin-fixed, paraffin-embedded sections of trachea, taken from chickens on days 3-10 post-inoculation (p.i.) with ILTV were hybridized...... on day 5 p.i. No hybridization was observed in 3 out of 3 chickens examined on day 10 p.i. ILTV nucleic acid was detected in nuclei of degenerated tracheal epithelial cells and in intranuclear inclusion bodies of syncytia....

  15. Effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids.

    Science.gov (United States)

    Bambou, J C; Archimède, H; Arquet, R; Mahieu, M; Alexandre, G; González-Garcia, E; Mandonnet, N

    2011-06-10

    The aim of the present study was to test the effect of dietary supplementation on resistance to experimental infection with Haemonchus contortus in Creole kids. One trial with three replicates involved a total of 154 female kids that were chosen from three successive cohorts of the Creole flock of INRA-Gardel in 2007. The kids were placed into four treatments according to the amount of concentrate they received: G0 (no concentrate and a quality Dichantium spp. hay ad libitum, HAY), G1 (HAY+100g commercial concentrate d(-1)), G2 (HAY+200 g commercial concentrate d(-1)), G3 (HAY+300 g commercial concentrate d(-1)). The G0-G3 groups were infected with a single dose of 10,000 H. contortus third stage larvae (L(3)) at Day 0 (D0). Each infected group was comprised of one half resistant and one half susceptible genetically indexed kids. The average breeding values on egg excretion at 11 months of age were distant of 0.70, 0.65, 0.61 and 0.61 genetic standard deviations in G0, G1, G2 and G3, respectively. The faecal egg count (FEC), packed cell volume (PCV), eosinophilia (EOSI) and dry matter intake (DMI) indices were monitored weekly until 42 days post-infection. Enzyme-linked immunosorbent assay was carried out on serum samples to determine the level of IgA anti-H. contortus L(3) crude extracts and adult excretion/secretion products (ESP). The 10,000 L(3) dose received by the kids induced a severe infection: 8000 eggs per gram at the FEC peak, a PCV less than 15% and mortality. Interestingly, the supplemented animals in G3 showed a higher level of EOSI but a lower level of IgA anti-L3 and IgA anti-ESP than non-supplemented animals (G0). Resistant and susceptible kids had significantly different FEC variations within the groups. Susceptible kids had a 1.6 times higher egg output than resistant kids in G0. This difference was not found in the supplemented groups. The results of this study showed that supplementary feeding improved resistance of Creole kids to H. contortus

  16. Toxoplasma gondii: infection natural congenital in cattle and an experimental inoculation of gestating cows with oocysts.

    Science.gov (United States)

    Costa, Gustavo Henrique Nogueira; da Costa, Alvimar José; Lopes, Welber Daniel Zanetti; Bresciani, Katia Denise Saraiva; dos Santos, Thais Rabelo; Esper, César Roberto; Santana, Aureo Evangelista

    2011-01-01

    Two studies, of a natural infection and an experimental infection, were performed in order to study congenital transmission of Toxoplasma gondii in cattle. In the first study, 50 fetuses were harvested from gestating cows that were eutanasied at a municipal slaughterhouse in Jaboticabal, São Paulo state, Brazil. In the second study, 11 gestating cows were divided into four groups for inoculation with T. gondii: GI consisted of three cows inoculated with 1.0 × 10(5) oocysts during their first trimester of gestation; GII consisted of three cows inoculated with 1.0 × 10(5) oocysts during their second trimester of gestation; GIII consisted of three cows inoculated with 1.0 × 10(5) oocysts during their last trimester of gestation; and GIV consisted of two control cows, one during its first and the other during its second trimester of gestation. In both studies, the presence of T. gondii was confirmed both indirectly by immunofluorescence assay (IFAT). In the natural infection experiment, 18% (9/50) of the gestating cows were confirmed to have specific antibodies (IFAT--1:64) against T. gondii. The bioassay was able to diagnose the presence of T. gondii in the tissue samples from three calves. In the second experiment, the nine cows from groups I, II and III presented with specific antibodies (IFAT) against T. gondii. In contrast, T. gondii could not be detected by IFAT, histopathological examination or the bioassay in any of the nine calves born to cows experimentally infected with T. gondii oocysts. Based on the results from both studies, we conclude that congenital infection of T. gondii in cattle, while infrequent, does occur naturally. The pathogenicity of the strain of T. gondii may influence the likelihood of this route of transmission. Copyright © 2010 Elsevier Inc. All rights reserved.

  17. Aspects of resistance to experimental infection with Trypanosoma cruzi

    International Nuclear Information System (INIS)

    Dias, Viviane Liotti

    2010-01-01

    Chagas disease, a zoonosis caused by the protozoan Trypanosoma cruzi, has a wide distribution in Latin America and extends from the southern part of the United States to Argentina. A number of 10 million of infected people is estimated and another 25 million exposed to the risk. Although discovered over a century, Chagas disease is still a serious infection that causes great socioeconomic impact, with no effective treatment at the chronic phase and in which, a lack of scientific knowledge can be observed. The main goal of this work was that obtaining and using consomic strain of mice, the resistance could be investigated. Consomic strains were produced by programmed mating, in which the animals were monitored with DNA polymorphic markers, and one of his chromosomes was replaced by his homologue from another strain. As parental, were used, the inbred strains C57BL/6/J Unib with resistant phenotype (donor) and as receiver, the A/JUnib strain, that has a susceptible phenotype. These models were used to produce five consomic strains: for the chromosomes 7 (CSs7), 11 (CSs11), 14 (CSs14), 17 (CSs17) and 19 (CSs19), described by Passos et al. (2003) as important in controlling infection caused by the Y strain of T. cruzi. In experimental testing, the consomics were inoculated intraperitoneally at doses of 10 1 , 10 2 , 10 3 and 10 4 using as control, animals from both parental lines. In all consomics, resistance was higher than that observed in the susceptible parental. In a second protocol, the consomics were mated with scheduled associations and the progenies were challenged with inocula employing increasing doses of trypomastigotes. The resistance observed in this group was also higher than that observed in the parental with susceptible phenotype. The observed results demonstrate that the use of the consomic strains that were produced order to assess the contribution of each chromosome in the resistance, as well as the effects of association between chromosomes are an

  18. Aberrant chlamydial developmental forms in the gastrointestinal tract of pigs spontaneously and experimentally infected with Chlamydia suis.

    Science.gov (United States)

    Pospischil, Andreas; Borel, Nicole; Chowdhury, Emdad H; Guscetti, Franco

    2009-03-16

    The phenomenon of persistence is well known from in vitro studies, where it is associated with the production of aberrant bodies, but its occurrence in vivo is less well documented. The objective of this study was to search for aberrant bodies in intestinal tissues from pigs, describe their ultrastructure, and investigate the suitability of immunohistochemical staining for chlamydial heat shock protein 60 (cHSP60) to detect such forms. Intestinal tissues derived from pigs naturally and experimentally infected with Chlamydia (C.) suis were examined by immunohistochemistry, transmission electron microscopy and immunogold electron microscopy. The chlamydial species involved in the natural infection were determined using an Array Tube Microarray to C. suis and Chlamydophila abortus. Ultrastructurally, aberrant bodies were detected in the gut of both naturally and experimentally infected pigs. Immunogold electron microscopy showed that the aberrant bodies were labeled less strongly than the normal forms by antibodies against LPS and cHSP60 respectively. It was concluded that aberrant bodies occur in vivo in pigs and that the gnotobiotic pig model might be suitable for the study of chlamydial persistence in vivo. The antibody against cHSP60 does not appear to be suitable to specifically detect such forms.

  19. Histopathological and parasitological investigations of ear healthy skin of dogs naturally and experimentally infected with Leishmania (Leishmania) chagasi.

    Science.gov (United States)

    Figueiredo, Maria Marta; Moura, Eliane Perlatto; Costa, Miriam Maria; Ribeiro, Vitor Marcio; Michalick, Marilene Suzan; Tafuri, Washington Luiz; Tafuri, Wagner Luiz

    2010-07-01

    Although 90% of clinical cases of American visceral leishmaniasis (AVL) occur in the northeastern region of Brazil, the incidence of cases in recent years has increased in southeastern states such as Minas Gerais (MG), where the disease has been reported in several cities, including Belo Horizonte, the state capital. Some studies have shown a strong correlation between the incidence of AVL and canine visceral leishmaniasis (CVL) in Belo Horizonte. A study of 108 dogs with parasite Leishmania chagasi detected by immuno-histochemistry in healthy ear skin was obtained from two distinct geographical areas: 55 from a metropolitan area of the municipality (Santa Luzia, MG) and 53 dogs from a central area of Belo Horizonte. In parallel, a group of 10 beagles were experimentally infected with L. chagasi. Considering the clinical aspects of all naturally infected dogs, symptomatic dogs were more frequent than asymptomatic ones, especially animals from the metropolitan area compared with the central area (79.6% and 20.3%, respectively). A chronic exudate was observed in the ear of 51 out of 55 dogs naturally infected from the metropolitan area (92.7%) and 45 out of 53 dogs naturally infected from the central area (84.9%). Importantly, asymptomatic dogs from the central area harbor more parasites in the skin than the asymptomatic ones from the metropolitan area. In addition, a profound difference was noted in the intensity of the inflammatory reaction and parasite load in the skin of experimental infected dogs.

  20. Efficiency of oxytetracycline treatment in rainbow trout experimentally infected with Flavobacterium psychrophilum strains having different in vitro antibiotic susceptibilities

    DEFF Research Database (Denmark)

    Bruun, Morten Sichlau; Madsen, Lone; Dalsgaard, Inger

    2003-01-01

    The medication effect of oxytetracycline on groups of rainbow trout fry experimentally infected with three strains of Flavobacterium psychrophilum was investigated. The infection model was based on intraperitoneal injection of the pathogen and treatment was done using medicated feed resulting...... in 100 mg oxytetracycline/kg fish for 10 days. The three F. psychrophilum strains had different antimicrobial susceptibilities and successful treatment was only obtained in the trial using a strain with a MICOTC of 0.25 mug/ml. No effect of treatment was seen in the group infected with a strain having...

  1. The Leishmania HSP20 Is Antigenic during Natural Infections, but, as DNA Vaccine, It does not Protect BALB/c Mice against Experimental L. amazonensis Infection

    Directory of Open Access Journals (Sweden)

    Ana M. Montalvo-Álvarez

    2008-01-01

    Full Text Available Protozoa of the genus Leishmania are causative agents of leishmaniasis, an important health problem in both human and veterinary medicine. Here, we describe a new heat shock protein (HSP in Leishmania, belonging to the small HSP (sHSP family in kinetoplastids. The protein is highly conserved in different Leishmania species, showing instead significant divergence with sHSP's from other organisms. The humoral response elicited against this protein during Leishmania infection has been investigated in natural infected humans and dogs, and in experimentally infected hamsters. Leishmania HSP20 is a prominent antigen for canine hosts; on the contrary, the protein seems to be a poor antigen for human immune system. Time-course analysis of appearance of anti-HSP20 antibodies in golden hamsters indicated that these antibodies are produced at late stages of the infection, when clinical symptoms of disease are patent. Finally, the protective efficacy of HSP20 was assessed in mice using a DNA vaccine approach prior to challenge with Leishmania amazonensis.

  2. Experimental treatment with sodium stibogluconate of hamsters infected with Leishmania (Leishmania) chagasi and Leishmania (Leishmania) amazonensis Tratamento experimental com stibogluconato de sódio em hamsters infectados com Leishmania (Leishmania) chagasi e Leishmania (Leishmania) amazonensis

    OpenAIRE

    Elizabeth M. de Figueiredo; Jaime Costa e Silva; Reginaldo P. Brazil

    1999-01-01

    The present paper reports the experimental treatment of hamsters infected with Leishmania chagasi and Leishmania amazonensis with sodium stibogluconate (20mg/kg/day x 20 days). Only with L. chagasi did the treatment result in the complete elimination of parasites from the spleen. However, no parasitological cure was achieved in hamsters infected with L. amazonensis.O presente trabalho é um relato do tratamento experimental de hamsters infectado com Leishmania chagasi e Leishmania amazonensis ...

  3. Experimental infection of snakes with Ophidiomyces ophiodiicola causes pathological changes that typify snake fungal disease

    Science.gov (United States)

    Lorch, Jeffrey M.; Lankton, Julia S.; Werner, Katrien; Falendysz, Elizabeth A.; McCurley, Kevin; Blehert, David S.

    2015-01-01

    Snake fungal disease (SFD) is an emerging skin infection of wild snakes in eastern North America. The fungus Ophidiomyces ophiodiicola is frequently associated with the skin lesions that are characteristic of SFD, but a causal relationship between the fungus and the disease has not been established. We experimentally infected captive-bred corn snakes (Pantherophis guttatus) in the laboratory with pure cultures of O. ophiodiicola. All snakes in the infected group (n = 8) developed gross and microscopic lesions identical to those observed in wild snakes with SFD; snakes in the control group (n = 7) did not develop skin infections. Furthermore, the same strain of O. ophiodiicola used to inoculate snakes was recovered from lesions of all animals in the infected group, but no fungi were isolated from individuals in the control group. Monitoring progression of lesions throughout the experiment captured a range of presentations of SFD that have been described in wild snakes. The host response to the infection included marked recruitment of granulocytes to sites of fungal invasion, increased frequency of molting, and abnormal behaviors, such as anorexia and resting in conspicuous areas of enclosures. While these responses may help snakes to fight infection, they could also impact host fitness and may contribute to mortality in wild snakes with chronic O. ophiodiicola infection. This work provides a basis for understanding the pathogenicity of O. ophiodiicola and the ecology of SFD by using a model system that incorporates a host species that is easy to procure and maintain in the laboratory.

  4. Behavioral and hormonal changes associated with the infective dose in experimental taeniasis in golden hamsters (Mesocricetus auratus).

    Science.gov (United States)

    Domínguez-Roldan, Rosa; Hallal-Calleros, Claudia; Sciutto, Edda; Hernández, Marisela; Aguirre-Flores, Virginio; García-Jiménez, Sara; Báez-Saldaña, Armida; Flores-Pérez, Fernando Iván

    2016-07-01

    It has been reported that behavioral changes relate to infection in different parasitoses. However, the relation between the extent of the behavioral changes and the magnitude of the infection has been scarcely studied. The aim of this study was to evaluate the relationship between different doses of infection and the behavioral changes induced in the experimental Taenia pisiformis taeniasis in golden hamsters. Groups of nine hamsters were infected with three or six T. pisiformis metacestodes. The locomotor activity was quantified daily in an open field test during the 21 days after infection; anxiety test was performed in an elevated plus-maze with a dark/light area at 7, 14 and 21 days post-infection, and serum cortisol levels were determined by radioimmunoassay before infection and at day 22 after infection. The challenge itself induced modifications on behavior and cortisol levels in hamsters, with or without successful infection (taenia development). Animals challenged with three metacestodes induced a decrease in locomotor activity and an increase in anxiety in infected animals. A higher and earlier decrease in locomotor activity and increased anxiety levels were observed in hamsters challenged with six cysticerci, which were accompanied by higher levels of sera cortisol at the end of the experiment. At necropsy, 44-55% of hamster became infected with an efficiency of implantation of 22-26%, challenged with three or six cysticerci respectively. The challenge of hamsters with metacestodes, promote behavioral changes in an extent dependent on the magnitude of the challenge, disregarding the effectiveness of the infection. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Influence of infection by Toxoplasma gondii on purine levels and E-ADA activity in the brain of mice experimentally infected mice.

    Science.gov (United States)

    Tonin, Alexandre A; Da Silva, Aleksandro S; Casali, Emerson A; Silveira, Stephanie S; Moritz, Cesar E J; Camillo, Giovana; Flores, Mariana M; Fighera, Rafael; Thomé, Gustavo R; Morsch, Vera M; Schetinger, Maria Rosa C; Rue, Mario De La; Vogel, Fernanda S F; Lopes, Sonia T A

    2014-07-01

    The aim of this study was to assess the purine levels and E-ADA activity in the brain of mice (BALB/c) experimentally infected with Toxoplasma gondii. In experiment I (n=24) the mice were infected with RH strain of T. gondii, while in experiment II (n=36) they were infected with strain ME-49 of T. gondii. Our results showed that, for RH strain (acute phase), an increase in both periods in the levels of ATP, ADP, AMP, adenosine, hypoxanthine, xanthine (only on day 6 PI) and uric acid (only on day 6 PI). By the other hand, the RH strain led, on days 4 and 6 PI, to a reduction in the concentration of inosine. ME-49, a cystogenic strain, showed some differences in acute and chronic phase, since on day 6 PI the levels of ATP and ADP were increased, while on day 30 these same nucleotides were reduced. On day 60 PI, ME-49 induced a reduction in the levels of ATP, ADP, AMP, adenosine, inosine and xanthine, while uric acid was increased. A decrease of E-ADA activity was observed in brain on days 4 and 6 PI (RH), and 30 PI (ME-49); however on day 60 PI E-ADA activity was increased for infection by ME-49 strain. Therefore, it was possible to conclude that infection with T. gondii changes the purine levels and the activity of E-ADA in brain, which may be associated with neurological signs commonly observed in this disease. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. Vaccine-mediated immune responses to experimental pulmonary Cryptococcus gattii infection in mice.

    Directory of Open Access Journals (Sweden)

    Ashok K Chaturvedi

    Full Text Available Cryptococcus gattii is a fungal pathogen that can cause life-threatening respiratory and disseminated infections in immune-competent and immune-suppressed individuals. Currently, there are no standardized vaccines against cryptococcosis in humans, underlying an urgent need for effective therapies and/or vaccines. In this study, we evaluated the efficacy of intranasal immunization with C. gattii cell wall associated (CW and/or cytoplasmic (CP protein preparations to induce protection against experimental pulmonary C. gattii infection in mice. BALB/c mice immunized with C. gattii CW and/or CP protein preparations exhibited a significant reduction in pulmonary fungal burden and prolonged survival following pulmonary challenge with C. gattii. Protection was associated with significantly increased pro-inflammatory and Th1-type cytokine recall responses, in vitro and increased C. gattii-specific antibody production in immunized mice challenged with C. gattii. A number of immunodominant proteins were identified following immunoblot analysis of C. gattii CW and CP protein preparations using sera from immunized mice. Immunization with a combined CW and CP protein preparation resulted in an early increase in pulmonary T cell infiltrates following challenge with C. gattii. Overall, our studies show that C. gattii CW and CP protein preparations contain antigens that may be included in a subunit vaccine to induce prolonged protection against pulmonary C. gattii infection.

  7. Gag- and env-specific serum antibodies in cats after natural and experimental infection with feline immunodeficiency virus.

    NARCIS (Netherlands)

    G.F. Rimmelzwaan (Guus); C.H.J. Siebelink (Kees); H. Broos; G.A. Drost; K. Weijer (Kees); R. van Herwijnen (Rob); A.D.M.E. Osterhaus (Albert)

    1994-01-01

    textabstractIn order to monitor the antibody response to feline immunodeficiency virus (FIV) in cats, following experimental and natural infection, enzyme-linked immunosorbent assays (ELISAs) were developed using recombinant env and gag proteins and p24-specific monoclonal antibodies. It was shown

  8. Experimental infection of Pacific oyster Crassostrea gigas spat by ostreid herpesvirus 1: demonstration of oyster spat susceptibility

    Directory of Open Access Journals (Sweden)

    Schikorski David

    2011-02-01

    Full Text Available Abstract In 2008 and 2009, acute mortalities occurred in France among Pacific cupped oyster, Crassostrea gigas, spat. Different hypothesis including the implication of environmental factors, toxic algae and/or pathogens have been explored. Diagnostic tests indicated that OsHV-1 including a particular genotype, termed OsHV-1 μVar, was detected in most of samples and especially in moribund oysters with the highlighting of virus particles looking like herpes viruses by TEM examination. In this study, an experimental protocol to reproduce OsHV-1 infection in laboratory conditions was developed. This protocol was based on the intramuscular injection of filtered (0.22 μm tissue homogenates prepared from naturally OsHV-1 infected spat collected on French coasts during mortality outbreaks in 2008. Results of the experimental trials showed that mortalities were induced after injection. Moreover, filtered tissue homogenates induced mortalities whereas the same tissue homogenates exposed to an ultraviolet (UV treatment did not induce any mortality suggesting that oyster spat mortalities require the presence of a UV sensitive agent. Furthermore, analysis of injected oyster spat revealed the detection of high amounts of OsHV-1 DNA by real-time quantitative PCR. Finally, TEM analysis demonstrated the presence of herpes virus particles. The developed protocol allowed to maintain sources of infective virus which can be useful for the development of further studies concerning the transmission and the development of OsHV-1 infection.

  9. Inhibiting fungal multidrug resistance by disrupting an activator-Mediator interaction.

    Science.gov (United States)

    Nishikawa, Joy L; Boeszoermenyi, Andras; Vale-Silva, Luis A; Torelli, Riccardo; Posteraro, Brunella; Sohn, Yoo-Jin; Ji, Fei; Gelev, Vladimir; Sanglard, Dominique; Sanguinetti, Maurizio; Sadreyev, Ruslan I; Mukherjee, Goutam; Bhyravabhotla, Jayaram; Buhrlage, Sara J; Gray, Nathanael S; Wagner, Gerhard; Näär, Anders M; Arthanari, Haribabu

    2016-02-25

    Eukaryotic transcription activators stimulate the expression of specific sets of target genes through recruitment of co-activators such as the RNA polymerase II-interacting Mediator complex. Aberrant function of transcription activators has been implicated in several diseases. However, therapeutic targeting efforts have been hampered by a lack of detailed molecular knowledge of the mechanisms of gene activation by disease-associated transcription activators. We previously identified an activator-targeted three-helix bundle KIX domain in the human MED15 Mediator subunit that is structurally conserved in Gal11/Med15 Mediator subunits in fungi. The Gal11/Med15 KIX domain engages pleiotropic drug resistance transcription factor (Pdr1) orthologues, which are key regulators of the multidrug resistance pathway in Saccharomyces cerevisiae and in the clinically important human pathogen Candida glabrata. The prevalence of C. glabrata is rising, partly owing to its low intrinsic susceptibility to azoles, the most widely used antifungal agent. Drug-resistant clinical isolates of C. glabrata most commonly contain point mutations in Pdr1 that render it constitutively active, suggesting that this transcriptional activation pathway represents a linchpin in C. glabrata multidrug resistance. Here we perform sequential biochemical and in vivo high-throughput screens to identify small-molecule inhibitors of the interaction of the C. glabrata Pdr1 activation domain with the C. glabrata Gal11A KIX domain. The lead compound (iKIX1) inhibits Pdr1-dependent gene activation and re-sensitizes drug-resistant C. glabrata to azole antifungals in vitro and in animal models for disseminated and urinary tract C. glabrata infection. Determining the NMR structure of the C. glabrata Gal11A KIX domain provides a detailed understanding of the molecular mechanism of Pdr1 gene activation and multidrug resistance inhibition by iKIX1. We have demonstrated the feasibility of small-molecule targeting of a

  10. Rapid and reliable MALDI-TOF mass spectrometry identification of Candida non-albicans isolates from bloodstream infections.

    Science.gov (United States)

    Pulcrano, Giovanna; Iula, Dora Vita; Vollaro, Antonio; Tucci, Alessandra; Cerullo, Monica; Esposito, Matilde; Rossano, Fabio; Catania, Maria Rosaria

    2013-09-01

    Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) fingerprinting has recently become an effective instrument for rapid microbiological diagnostics and in particular for identification of micro-organisms directly in a positive blood culture. The aim of the study was to evaluate a collection of 82 stored yeast isolates from bloodstream infection, by MALDI-TOF MS; 21 isolates were identified also directly from positive blood cultures and in the presence of other co-infecting micro-organisms. Of the 82 isolates grown on plates, 64 (76%) were correctly identified by the Vitek II system and 82 (100%) by MALDI-TOF MS; when the two methods gave different results, the isolate was identified by PCR. MALDI-TOF MS was unreliable in identifying two isolates (Candida glabrata and Candida parapsilosis) directly from blood culture; however, direct analysis from positive blood culture samples was fast and effective for the identification of yeast, which is of great importance for early and adequate treatment. © 2013. Published by Elsevier B.V. All rights reserved.

  11. The effect of a fenbendazole treatment on cyst excretion and weight gain in calves experimentally infected with Giardia duodenalis.

    Science.gov (United States)

    Geurden, Thomas; Vandenhoute, Els; Pohle, Herbert; Casaert, Stijn; De Wilde, Nathalie; Vercruysse, Jozef; Claerebout, Edwin

    2010-04-19

    A total of 28 Holstein-Friesian calves were experimentally infected with 10(5)Giardia duodenalis cysts. Eleven days later, all animals were allocated into two groups of 14 animals each, based on the average pre-treatment cyst counts. Treatment was randomly assigned to one of the two groups, and all animals in the treatment group received a daily oral dosage of 15mg fenbendazole per kg bodyweight during 3 consecutive days. The calves in the control group received a placebo (water). From 3 days after treatment onwards, cyst excretion was determined three times a week during 4 consecutive weeks. The faecal consistency and general health were recorded on a daily basis, and all animals were weighed prior to treatment and weekly thereafter. At the end of the experimental period, there was a significant (Pfenbendazole treated and untreated calves experimentally infected with G. duodenalis, although additional data need to confirm the need for treatment in natural conditions.

  12. EFFECT OF HIGHLY ACTIVE ANTIRETROVIRAL THERAPY ON VAGINAL Candida spp. ISOLATION IN HIV-INFECTED COMPARED TO HIV-UNINFECTED WOMEN

    Directory of Open Access Journals (Sweden)

    Silvia de Souza Dantas ALCZUK

    2015-04-01

    Full Text Available Vulvovaginal candidiasis (VVC in HIV-infected women contributed to the impairment of their quality of life. The aim of this study was to evaluate the effect of highly active antiretroviral therapy (HAART use on the vaginal Candida spp. isolation in HIV-infected compared to HIV-uninfected women. This cross-sectional study included 178 HIV-infected (HIV group and 200 HIV-uninfected women (control that were studied at the Specialized Assistance Service (SAE for sexually transmitted diseases (STD/AIDS of the city of Maringá, Brazil, from April 1 to October 30, 2011. The yeasts were isolated and identified by phenotypic and molecular methods. The in vitro antifungal susceptibility to fluconazole, itraconazole, nystatin and amphotericin B was tested by the reference microdilution method. Higher frequencies of total vaginal Candida spp. isolation were found in the HIV-infected group than in the control group. However, both groups showed a similar frequency of colonization and VVC. Although C. albicans was the most frequent and sensitive to azolics and polyenes in both HIV-infected and uninfected women, the emerging resistance of C. glabrata to amphotericin B in the HIV-infected women was observed. Although higher frequency of vaginal Candida spp. isolation had been observed in the HIV-infected than in HIV-uninfected women, colonization and VVC showed similar frequency in both groups, indicating that HAART appears to protect against vaginal colonization and VVC.

  13. Experimental Mycoplasma gallisepticum infections in captive-reared wild turkeys

    Science.gov (United States)

    Rocke, Tonie E.; Yuill, Thomas M.; Amundson, Terry E.

    1988-01-01

    The effects of Mycoplasma gallisepticum (MG) infections on egg production, fertility, and hatchability were studied in captive-reared wild turkeys (Meleagris gallopavo). Three groups of adult birds, each consisting of four hens and two toms, were exposed to MG by the respiratory route at the beginning of their breeding season. Fourteen control birds received sterile growth medium. Although no mortality of infected or control birds occurred, egg production during the first breeding season after infection was reduced. The mean number of eggs/hen/day produced by infected groups the first breeding season postexposure (PE) was significantly lower than the control value. The mean number of eggs produced daily by the same hens 1 yr later was unaffected by MG infection. The pecentage of fertile eggs produced by infected groups was slightly reduced in both the first and second breeding seasons PE. Hatchability of fertile eggs from infected hens was significantly lower than eggs from control hens. Productivity may be impaired if MG infections occur in free-ranging wild turkey populations.

  14. Lactobacillus paracasei feeding improves the control of secondary experimental meningococcal infection in flu-infected mice.

    Science.gov (United States)

    Belkacem, Nouria; Bourdet-Sicard, Raphaëlle; Taha, Muhamed-Kkeir

    2018-04-10

    The use of probiotics to improve anti-microbial defence, such as for influenza infections, is increasingly recommended. However, no data are available on the effect of probiotics on flu-associated secondary bacterial infections. There is strong evidence of a spatiotemporal association between influenza virus infection and invasive Neisseria meningitidis. We thus investigated the effect of feeding mice Lactobacillus paracasei CNCM I-1518 in a mouse model of sequential influenza-meningococcal infection. We intranasally infected BALB/c mice with a strain of influenza A virus (IAV) H3N2 that was first adapted to mice. Seven days later, a secondary bacterial infection was induced by intranasal administration of bioluminescent N. meningitidis. During the experiment, mice orally received either L. paracasei CNCM I-1518 or PBS as a control. The effect of L. paracasei administration on secondary bacterial infection by N. meningitidis was evaluated. Oral consumption of L. paracasei CNCM I-1518 reduced the weight loss of infected mice and lowered the bioluminescent signal of infecting meningococci. This improvement was associated with higher recruitment of inflammatory myeloid cells, such as interstitial monocytes and dendritic cells, to the lungs. Our data highlight the role of the gut-lung axis. L. paracasei CNCM I-1518 may boost the defence against IAV infection and secondary bacterial infection, which should be further studied and validated in clinical trials.

  15. Experimental study of European bat lyssavirus type-2 infection in Daubenton's bats (Myotis daubentonii).

    Science.gov (United States)

    Johnson, Nicholas; Vos, Ad; Neubert, Larissa; Freuling, Conrad; Mansfield, Karen L; Kaipf, Ingrid; Denzinger, Annette; Hicks, Dan; Núñez, Alex; Franka, Richard; Rupprecht, Charles E; Müller, Thomas; Fooks, Anthony R

    2008-11-01

    European bat lyssavirus type 2 (EBLV-2) can be transmitted from Daubenton's bats to humans and cause rabies. EBLV-2 has been repeatedly isolated from Daubenton's bats in the UK but appears to be present at a low level within the native bat population. This has prompted us to investigate the disease in its natural host under experimental conditions, to assess its virulence, dissemination and likely means of transmission between insectivorous bats. With the exception of direct intracranial inoculation, only one of seven Daubenton's bats inoculated by subdermal inoculation became infected with EBLV-2. Both intramuscular and intranasal inoculation failed to infect the bats. No animal inoculated with EBLV-2 seroconverted during the study period. During infection, virus excretion in saliva (both viral RNA and live virus) was confirmed up to 3 days before the development of rabies. Disease was manifested as a gradual loss of weight prior to the development of paralysis and then death. The highest levels of virus were measured in the brain, with much lower levels of viral genomic RNA detected in the tongue, salivary glands, kidney, lung and heart. These observations are similar to those made in naturally infected Daubenton's bats and this is the first documented report of isolation of EBLV-2 in bat saliva. We conclude that EBLV-2 is most likely transmitted in saliva by a shallow bite.

  16. Efficacy of one dose vaccination against experimental infection with two Mycoplasma hyopneumoniae strains.

    Science.gov (United States)

    Michiels, Annelies; Arsenakis, Ioannis; Boyen, Filip; Krejci, Roman; Haesebrouck, Freddy; Maes, Dominiek

    2017-08-29

    Mycoplasma hyopneumoniae (M. hyopneumoniae) is the primary agent of enzootic pneumonia in pigs. Pigs are often infected with different M. hyopneumoniae strains. This study assessed the efficacy of vaccination against experimental infection with two genetically different M. hyopneumoniae strains in weaned piglets. At 33 days of age (D0), 45 M. hyopneumoniae-free piglets were randomly assigned to three different groups: 1) negative control group (NCG; n = 5): not vaccinated, not infected, 2) positive control group (PCG; n = 20): not vaccinated, infected, and 3) vaccination group (VG; n = 20): single vaccination with an inactivated whole-cell M. hyopneumoniae vaccine (Hyogen®, Ceva) (D1), infected. The PCG and VG were endotracheally inoculated with 7 × 10 7 CCU in 7 ml of the highly virulent M. hyopneumoniae strain F7.2C (D24) and 7 × 10 7 CCU in 7 ml low virulent strain F1.12A (D25). A respiratory disease score (RDS) was assessed from D24 until D53. At D53 (euthanasia), macroscopic lung lesions (MLL) were scored, log copies of M. hyopneumoniae DNA (qPCR) and IL-1 and IL-6-concentrations (ELISA) on bronchoalveolar lavage fluid were determined. The RDS and MLL at euthanasia were respectively 0, 1.20 and 0.55 (P hyopneumoniae strain as the vaccinated pigs coughed significantly less, and showed significantly less lung lesions compared to the non-vaccinated challenged pigs: the vaccinated animals showed a 52.9% lower RDS and 91.0% lower MLL compared to the PCG. In the bronchoalveolar lavage fluid collected at the necropsy of the vaccinated pigs, a significantly lower amount of M. hyopneumoniae-DNA and a significantly lower IL-1 and IL-6 concentration was found compared to the pigs of the PCG.

  17. Experimental infection of Aphanomyces invadans and susceptibility in seven species of tropical fish

    Directory of Open Access Journals (Sweden)

    Seyedeh F. Afzali

    2015-09-01

    Full Text Available Aim: Epizootic ulcerative syndrome (EUS causes by aquatic oomycete fungus, Aphanomyces invadans is a dangerous fish disease of a wide range of fresh and brackish water, wild and farmed fish throughout the world. The objective of the present study was to determine the susceptibility of a number of tropical fish species to the EUS and compare the severity of infection between experimental groups. Materials and Methods: Snakehead, Channa striata (Bloch, 1793; snakeskin gourami, Trichopodus pectoralis (Regan, 1910; koi carp, Cyprinus carpio (Linnaeus, 1758; broadhead catfish, Clarias macrocephalus (Günther, 1864; goldfish, Carassius auratus (Linnaeus, 1758; climbing perch, Anabas testudineus (Bloch, 1792; and Nile tilapia, Oreochromis niloticus (Linnaeus, 1758 were challenged by intramuscular injection using zoospores of Aphanomyces invadans (NJM9701. The infected fish skins and muscles were examined for EUS histopathological characteristics, and the results on the severity of lesions and mortality were analyzed using SPSS program. Results: All zoospore-injected fish were shown to be susceptible to the EUS infection except Nile tilapia. Although, the general histopathological pattern was similar in the zoospore-injected group, but there were some variation in granulomatous reaction, that is the presence or absence of giant cells, and time of mortality were detected. The result of statistical analysis showed that there was a significant difference between species, (c2=145.11 and p<0.01. Conclusion: Gourami, koi carp, and catfish were demonstrated to be highly susceptible while goldfish and climbing perch were found to be moderately susceptible to the EUS infection. These findings suggested that the cellular response of fish to mycotic infection and granulomatous reaction varied in different fish species, which could not be an indicator of susceptibility or resistant to the EUS itself, although it was shown that the granulation rate and the level of

  18. Molecular detection of candida species from hospitalized patient’s specimens.

    Science.gov (United States)

    Camacho-Cardoso, José Luis; Martínez-Rivera, María Ángeles; Manzano-Gayosso, Patricia; Méndez-Tovar, Luis Javier; López-Martínez, Rubén; Hernández-Hernández, Francisca

    To identify the most frequent Candida species in specimens from patients hospitalized in different medical centers of Mexico City, with suspected fungal infection. Specimens were grown on Sabouraud dextrose agar at 28°C for 72 h. In addition, DNA was extracted. Isolates were grown on CHROMagar Candida™, at 37°C for 48 h. The molecular identification was performed by polymerase chain reaction (PCR) using primers specific for four species. Eighty one specimens were processed and included: bronchial lavage, pleural, cerebrospinal, peritoneal, ascites and bile fluids; blood, sputum, bone marrow, oro-tracheal cannula and ganglion. By culture, 30 samples (37%) were positive, and by PCR, 41 (50.6%). By PCR, the frequency of species was: Candida albicans 82.9%, Candida tropicalis 31.7%, Candida glabrata 24.4%, and Candida parapsilosis 4.9%. In 34.1% of specimens a species mixture was detected suggesting a co-infection: Two species in five specimens (C. albicans-C tropicalis and C. albicans-C glabrata), and three species in three specimens (C. albicans-C. glabrata-C. tropicalis). The PCR is an useful tool for detection the most common Candida species causing infection in hospitalized patients, it avoids the requirement of culture weather we start from clinical specimen and it favors the early diagnosis of invasive candidiasis. Copyright: © 2017 SecretarÍa de Salud

  19. Tupaia belangeri as an experimental animal model for viral infection.

    Science.gov (United States)

    Tsukiyama-Kohara, Kyoko; Kohara, Michinori

    2014-01-01

    Tupaias, or tree shrews, are small mammals that are similar in appearance to squirrels. The morphological and behavioral characteristics of the group have been extensively characterized, and despite previously being classified as primates, recent studies have placed the group in its own family, the Tupaiidae. Genomic analysis has revealed that the genus Tupaia is closer to humans than it is to rodents. In addition, tupaias are susceptible to hepatitis B virus and hepatitis C virus. The only other experimental animal that has been demonstrated to be sensitive to both of these viruses is the chimpanzee, but restrictions on animal testing have meant that experiments using chimpanzees have become almost impossible. Consequently, the development of the tupaia for use as an animal infection model could become a powerful tool for hepatitis virus research and in preclinical studies on drug development.

  20. Biochemical studies in experimentally Escherichia coli infected broiler chicken supplemented with neem (Azadirachta indica leaf extract

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    Vikash Sharma

    2015-11-01

    Full Text Available Aim: An experimental study was conducted on 192-day-old broiler chicks for evaluating the effect of 10% neem leaf extract (NLE supplementationon biochemical parameters in chickens experimentally infected with Escherichia coli O78 at 107 CFU/0.5 ml at 7 days of age. Materials and Methods: The 192-day-old broiler chicks were procured. These chicks were divided into two groups (A and B containing 96 birds each on the 1st day. Diet of all the chicks of Group A was supplemented with 10%NLE in water, whereas chicks of Group B were given feed and water devoid of NLE supplementation throughout the experiment. After rearing for 1 week, chicks of both the groups (A and B were again divided into two subgroups (Group A into A1 and A2 and Group B into B1 and B2 of 54 and 42 birds, respectively. At the age of 7 days all the chicks of groups A1 and B1 were injected with E. coli O78 at 107 CFU/0.5 ml intraperitoneally. Blood samples were collected from six chicks from each group at day 0, 2, 4, 7, 14, 21, 28 days post-infection and serum was separated for biochemical studies. Results: There was a significant increase in serum alanine transaminase (ALT, aspartate transaminase (AST, lactate dehydrogenase (LDH activities, globulin concentration and a decrease in total protein (TP, albumin concentrations, and alkaline phosphatase (ALP activity in both the infected groups. However, the changes in biochemical values, i.e., ALT, AST, LDH, ALP, TP, albumin, and globulin wereof lower magnitude in NLE supplemented group suggesting hepatoprotective and cardioprotective effect of NLE. Conclusions: Fromthe present study, it is reasonable to conclude that significant increase in the value of ALT, AST, LDH, globulin, and significant decrease in the value of ALP, TP, and albumin was of lower magnitude in supplemented infected group (A1 as compared to non-supplemented infected group (B1 suggesting hepatoprotective and cardioprotective effect of NLE.

  1. Candida-induced prosthetic joint infection. A literature review including 72 cases and a case report.

    Science.gov (United States)

    Cobo, Fernando; Rodríguez-Granger, Javier; López, Enrique M; Jiménez, Gemma; Sampedro, Antonio; Aliaga-Martínez, Luis; Navarro-Marí, José María

    2017-02-01

    The clinical and microbiological characteristics of prosthetic joint infection (PJI) caused by Candida species is described, including 72 cases in the literature and a case of Candida glabrata infection handled at the present centre. We describe one patient and using the key words 'fungal prosthetic joint infection' and 'candida prosthetic joint infection' we searched MEDLINE (National Library of Medicine, Bethesda, MD), Web of Science, CINAHL and Cochrane systematic review databases for case reports of this condition. Out of the 73 patients, 38 were female; mean age at diagnosis was 65.7 (± SD 18) yrs; 50 had risk factors for candidal infection such as systemic disease (e.g. rheumatoid arthritis, Sjogren's syndrome, systemic lupus erythematosus) and/or immunosuppressive therapy in 18 (24.6%) cases, diabetes mellitus in 14 (19.1%), immunosuppression due to malignant or chronic disease in 24 (32.8%) and long-term antibiotic use in four (5.4%) patients. Infection site was the knee in 36 patients and hip in 35; pain was present in 43 patients and swelling in 23 and the mean surgery-diagnosis interval was 32 months. The most frequent species was C. albicans, followed by C. parapsilosis. The diagnosis was obtained from joint fluid aspirate in 33 cases and intra-operative samples in 16. Susceptibility to antifungals was tested in only 21 isolates. The most frequently used antifungals were fluconazole and amphotericin B. Two-stage exchange arthroplasty was performed in 30 patients and resection arthroplasty in 31; 56 patients were cured with a combination of medical and surgical treatment; one patient died from the infection. PJI caused by Candida requires a high index of suspicion; surgery with long-term antifungal therapy is recommended.

  2. Experimental infection of Balb/c nude mice with Hepatitis E virus

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    Zhu Jianguo

    2009-06-01

    Full Text Available Abstract Background Several animal species can reportedly act as reservoirs for Hepatitis E virus (HEV, a zoonotic pathogen. HEV and antibody to the virus have been detected in a variety of animals including rodents. Pig and rat models for HEV have been established for HEV, but a nude mouse has not yet been developed. Methods Balb/c nude mice were inoculated with swine HEV, both orally and via intravenous injection to insure infection. Negative control and experimental contact-exposed groups of mice were also included in the study. The liver, spleen, kidney, jejunum, ileum, cecum and colon of each mouse from all three groups were collected for reverse transcription nested polymerase chain reaction (RT-nPCR detection, indirect immunofluorescence observation and histopathologic examination. The sera from nude mice were tested for anti-HEV IgG by enzyme linked immunosorbent assay (ELISA. Activities of liver enzymes, including alanine aminotransferase (ALT, aspartate aminotransferase (AST and alkaline phosphatase (ALP, as well as total bilirubin (TBIL were also measured in the sera of the nude mice. Results HEV antigens and HEV RNA were detected in liver, spleen, kidney, jejunum, ileum and colon both by indirect immunofluorescence and by RT-nPCR in all of the inoculated and in one of the contact-exposed nude mice. Histopathological changes were observed in the liver and spleen of these mice. Infected mice showed increased levels of AST, ALP, and anti-HEV IgG in sera. The livers of contact-exposed mice showed obvious histopathological damage. Conclusion Nude mice could be readily infected by HEV isolated from pigs. The nude mouse may therefore be a useful animal model for studying the pathogenesis of HEV.

  3. Diagnostic performances of two rapid tests for detection of feline leukemia virus antigen in sera of experimentally feline leukemia virus-infected cats

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    Matthew R Krecic

    2017-12-01

    Full Text Available Objectives The objective of this study was to compare the diagnostic sensitivities and specificities of WITNESS FeLV-FIV (Zoetis and SNAP FIV/FeLV Combo Test (IDEXX for the detection of FeLV p27 antigen in the sera of experimentally feline leukemia virus (FeLV-infected cats. Methods Diagnostic sensitivities of WITNESS and SNAP were determined through testing of 47 serum samples collected from cats day 56 post-experimental infection with a virulent FeLV Rickard strain. Successful experimental infection was confirmed based on observation of FeLV antigen and proviral DNA in anti-coagulated (EDTA whole-blood samples by immunofluorescent antibody (IFA test and PCR, respectively. Diagnostic specificities of both tests were determined through testing of sera of 92 laboratory-housed, non-FeLV-exposed specific pathogen-free (SPF cats. Results Forty-one of 47 blood samples were IFA positive, whereas all 47 samples were PCR positive. All 92 non-FeLV-infected SPF cats were IFA and PCR negative. In comparison to IFA as the reference method, both WITNESS and SNAP tests yielded equivalent sensitivities and specificities of 100% and 97.8%, respectively. In comparison to PCR as the reference method, both WITNESS and SNAP tests likewise performed equivalently, with sensitivities and specificities of 91.5% and 100%, respectively. Conclusions and relevance Sensitivity and specificity of WITNESS FeLV-FIV for identifying FeLV p27 antigen in the sera of these experimentally FeLV-infected and non-FeLV-exposed SPF cats equaled those of the SNAP FIV/FeLV Combo Test. However, all positive results, regardless of the point-of-care test used, should be confirmed before making clinical decisions such as segregation from other cats or euthanasia.

  4. Temporal dynamics of 'HoBi'-like pestivirus quasispecies in persistently infected calves generated under experimental conditions.

    Science.gov (United States)

    Weber, Matheus N; Bauermann, Fernando V; Canal, Cláudio W; Bayles, Darrell O; Neill, John D; Ridpath, Julia F

    2017-01-02

    'HoBi'-like virus is an atypical group within the Pestivirus genus that is implicated in economic losses for cattle producers due to both acute and persistent infections. Pestivirus strains exist as quasispecies (swarms of individual viruses) in infected animals and the viral populations making up the quasispecies differ widely in size and diversity in each animal. In the present study the viral quasispecies circulating in persistently infected (PI) calves, generated and maintained under experimental conditions using two different 'HoBi'-like strains, was observed over time. An increase in genetic variability and the development of certain mutations was observed over time. Mutations observed included the loss of a putative N-linked glycosylation site in the E2 region and the change of specific residues in E1/E2. It is hypothesized that these changes may be the results on continued adaption of the pestivirus to individual hosts. This is the first study characterizing variation in the viral swarms of animals persistently infected with HoBi-like viruses over time. Studies of the shifts in PI viral swarms will contribute to our understanding of the host and viral mechanisms that function in the maintenance of pestivirus persistent infections. Published by Elsevier B.V.

  5. Pathogenesis and Immunohistochemical Studies of Caprine Pleuropneumonia in Experimentally Infected Goats

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    Umer Sadique*, Zafar Iqbal Chaudhry1, Muhammad Younus Rana2, Aftab Ahmad Anjum3, Zahoor-Ul-Hassan, Abdul Sajid and Muhammad Mushtaq

    2012-06-01

    Full Text Available This study was designed to evaluate the pathogenesis of caprine pleuropneumonia (CPP in the experimentally inoculated goats with Mycoplasma mycoides subspecies Capri (Mmc. For this purpose, 12 goats (Group B were inoculated with bacterial isolates of Mmc while four goats were kept as untreated control (Group A. Clinical signs of the disease were recorded twice daily. Two goats from group B were sacrificed on weekly basis to demonstrate gross pathological lesions in different organs. Tissue samples from lungs, trachea, liver, heart, kidney, spleen, and small intestines were preserved for histopathological studies. The lungs and lymph nodes were preserved to demonstrate the antigen in tissue by using immuno- histochemical technique. The disease was successfully reproduced in all infected goats with severe manifestation. The clinical signs and gross lesions of the disease were mild at the beginning and became severe at the third and fourth weeks and then progressed to moderate and chronic forms. The histopathological lesions characteristic of CPP were found in all the organs. Antigen of Mmc was detected in tissue sections of lungs and lymph nodes. In conclusion, the disease was efficiently reproduced in experimental animals that showed acute septicemic form with lethal outcome.

  6. Experimental Infection of Snakes with Ophidiomyces ophiodiicola Causes Pathological Changes That Typify Snake Fungal Disease.

    Science.gov (United States)

    Lorch, Jeffrey M; Lankton, Julia; Werner, Katrien; Falendysz, Elizabeth A; McCurley, Kevin; Blehert, David S

    2015-11-17

    Snake fungal disease (SFD) is an emerging skin infection of wild snakes in eastern North America. The fungus Ophidiomyces ophiodiicola is frequently associated with the skin lesions that are characteristic of SFD, but a causal relationship between the fungus and the disease has not been established. We experimentally infected captive-bred corn snakes (Pantherophis guttatus) in the laboratory with pure cultures of O. ophiodiicola. All snakes in the infected group (n = 8) developed gross and microscopic lesions identical to those observed in wild snakes with SFD; snakes in the control group (n = 7) did not develop skin infections. Furthermore, the same strain of O. ophiodiicola used to inoculate snakes was recovered from lesions of all animals in the infected group, but no fungi were isolated from individuals in the control group. Monitoring progression of lesions throughout the experiment captured a range of presentations of SFD that have been described in wild snakes. The host response to the infection included marked recruitment of granulocytes to sites of fungal invasion, increased frequency of molting, and abnormal behaviors, such as anorexia and resting in conspicuous areas of enclosures. While these responses may help snakes to fight infection, they could also impact host fitness and may contribute to mortality in wild snakes with chronic O. ophiodiicola infection. This work provides a basis for understanding the pathogenicity of O. ophiodiicola and the ecology of SFD by using a model system that incorporates a host species that is easy to procure and maintain in the laboratory. Skin infections in snakes, referred to as snake fungal disease (SFD), have been reported with increasing frequency in wild snakes in the eastern United States. While most of these infections are associated with the fungus Ophidiomyces ophiodiicola, there has been no conclusive evidence to implicate this fungus as a primary pathogen. Furthermore, it is not understood why the

  7. Identification and Determination of Drug Resistant of Candida species isolated from Hospital Acquired Infections

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    Kambiz Diba

    2015-01-01

    Full Text Available Background & aim: Currently, the use of antifungal azole group and yeasts resistant to these drugs is increasing. The aim of this study was to isolate and identify the yeasts obtained from candidiasis patients and furthermore determining thier antifungal resistance. Methods: In the present descriptive study, infections samples were collected from 256 patients with suspected nosocomial candidiasis, then direct exam and culture were performed. Yeast colonies were identified using phenotypic methods, polymerase chain reaction method and enzyme digestion. Data were analyzed using Descriptive statistical tests. Results: Of sixty isolated yeast, thirty-seven cases of Candida albicans (61.6%, seven cases of C. krusei and C. glabrata (11.6% each, five cases of C. dubliniensis (8.3% and four cases of C. tropicalis (6.6% were indicated. The study showed that the sensitivity of C. albicans and C. cruise species to amphotericin B was negligible in disk diffusion and very sensitve in microdilution. Conclusion: Inspite of the results of antifungal susceptibility test of strains studied did not show high resistance, but screening for drug-resistant Candida isolates in Candida infection by disk diffusion and microdilution methods for new cases of drug resistance is reasonable.

  8. Cytokine, antibody and proliferative cellular responses elicited by Taenia solium calreticulin upon experimental infection in hamsters.

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    Fela Mendlovic

    Full Text Available Taenia solium causes two diseases in humans, cysticercosis and taeniosis. Tapeworm carriers are the main risk factor for neurocysticercosis. Limited information is available about the immune response elicited by the adult parasite, particularly the induction of Th2 responses, frequently associated to helminth infections. Calreticulin is a ubiquitous, multifunctional protein involved in cellular calcium homeostasis, which has been suggested to play a role in the regulation of immune responses. In this work, we assessed the effect of recombinant T. solium calreticulin (rTsCRT on the cytokine, humoral and cellular responses upon experimental infection in Syrian Golden hamsters (Mesocricetus auratus. Animals were infected with T. solium cysticerci and euthanized at different times after infection. Specific serum antibodies, proliferative responses in mesenteric lymph nodes and spleen cells, as well as cytokines messenger RNA (mRNA were analyzed. The results showed that one third of the infected animals elicited anti-rTsCRT IgG antibodies. Interestingly, mesenteric lymph node (MLN cells from either infected or non-infected animals did not proliferate upon in vitro stimulation with rTsCRT. Additionally, stimulation with a tapeworm crude extract resulted in increased expression of IL-4 and IL-5 mRNA. Upon stimulation, rTsCRT increased the expression levels of IL-10 in spleen and MLN cells from uninfected and infected hamsters. The results showed that rTsCRT favors a Th2-biased immune response characterized by the induction of IL-10 in mucosal and systemic lymphoid organs. Here we provide the first data on the cytokine, antibody and cellular responses to rTsCRT upon in vitro stimulation during taeniasis.

  9. Experimental study of action of autostrains Aerococcus viridans on the model Pseudomonas infection

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    D. O. Stepanskyi

    2017-05-01

    Full Text Available The paper presents the results of a study of the action of Aerococcus autostrains on the model of a chronic blue pus infection. For the study of the action of Aerococcus autosymbiont strains on Pseudomonas aeruginosa, three of the most biochemically and antagonistically active isolates were selected: 1 5m2015 (isolated from mice; 2 3k2015 (isolated from rats; 3 3ch2015 (isolated from humans. Experiments were conducted on 84 white outbred mice weighing 16–17 g, 60 were used as the experimental, and 24 as the control group. In the experimental group of animals, infected wounds were treated by Aerococcus autosymbiont strains once daily (0.2 billion ml–1 till recovery. The drug was administered under the scab with a syringe. In the control animals the wound was treated by isotonic sodium chloride solution (concentration 0.9% with the same route of administration and for the same period of time. It was found that from the very first days of application of Aerococci autosymbiont strains, perifocal inflammation was less severe in most animals in the research group compared with the control group. Starting from the fourth day of usage of Aerococcus autosymbiont strains the number of pseudomonades, contained in secretions from wounds in the experimental group of mice was significantly lower than in the control animals. It was revealed that in case of application of Aerococcus strain (5m2015 isolated from mice, the animals had better indicators of recovery, dynamics of local clinical signs of inflammation and the number of pseudomonades contained in the wound in comparison with other Aerococcus autostrains isolated from rats and humans. The wounds purified from pus and covered with dry scab faster. For example, wounds completely healed with dry scab rejection by the 11th day of observation in 44 of 58 surviving mice (75.9%. In the control group a similar pattern was observed in only 3 of 17 mice (17.6% by that period. The number of Pseudomonas

  10. Influence of experimental distemper infection on the distribution of lead in dogs previously subacutely intoxicated with lead carbonate

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    White, D.J.; Marshall, A.J.; McLeod, S.

    1975-01-01

    The ability of experimental canine distemper infection to mobilize body lead deposits has been studied in Beagle dogs previously subacutely intoxicated with lead carbonate. For comparative purposes dogs were included which had either received lead only or distemper only or remained undosed. It was found that in dogs predosed with lead, distemper infection resulted in a significant increase in lead levels in blood and urine; this coincided with the peak body temperatures reached on the third day post infection. It was also found that the lead content of the liver and bone of these dogs was considerably higher than that of dogs receiving lead alone; at the same time bone phosphorus showed a marked decrease while bone calcium values remained similar to undosed controls.

  11. DNA immunization against experimental genital herpes simplex virus infection.

    Science.gov (United States)

    Bourne, N; Stanberry, L R; Bernstein, D I; Lew, D

    1996-04-01

    A nucleic acid vaccine, expressing the gene encoding herpes simplex virus (HSV) type 2 glycoprotein D (gD2) under control of the cytomegalovirus immediate-early gene promoter, was used to immunize guinea pigs against genital HSV-2 infection. The vaccine elicited humoral immune responses comparable to those seen after HSV-2 infection. Immunized animals exhibited protection from primary genital HSV-2 disease with little or no development of vesicular skin lesions and significantly reduced HSV-2 replication in the genital tract. After recovery from primary infection, immunized guinea pigs experienced significantly fewer recurrences and had significantly less HSV-2 genomic DNA detected in the sacral dorsal root ganglia compared with control animals. Thus, immunization reduced the burden of latent infection resulting from intravaginal HSV-2 challenge, and a nucleic acid vaccine expressing the HSV-2 gD2 antigen protected guinea pigs against genital herpes, limiting primary infection and reducing the magnitude of latent infection and the frequency of recurrent disease.

  12. Candidiasis: a fungal infection--current challenges and progress in prevention and treatment.

    Science.gov (United States)

    Hani, Umme; Shivakumar, Hosakote G; Vaghela, Rudra; Osmani, Riyaz Ali M; Shrivastava, Atul

    2015-01-01

    Despite therapeutic advances candidiasis remains a common fungal infection most frequently caused by C. albicans and may occur as vulvovaginal candidiasis or thrush, a mucocutaneous candidiasis. Candidiasis frequently occurs in newborns, in immune-deficient people like AIDS patients, and in people being treated with broad spectrum antibiotics. It is mainly due to C. albicans while other species such as C. tropicalis, C. glabrata, C. parapsilosis and C. krusei are increasingly isolated. OTC antifungal dosage forms such as creams and gels can be used for effective treatment of local candidiasis. Whereas, for preventing spread of the disease to deeper vital organs, candidiasis antifungal chemotherapy is preferred. Use of probiotics and development of novel vaccines is an advanced approach for the prevention of candidiasis. Present review summarizes the diagnosis, current status and challenges in the treatment and prevention of candidiasis with prime focus on host defense against candidiasis, advancements in diagnosis, probiotics role and recent progress in the development of vaccines against candidiasis.

  13. Sarcocystis neurona infections in raccoons (Procyon lotor): evidence for natural infection with sarcocysts, transmission of infection to opossums (Didelphis virginiana), and experimental induction of neurologic disease in raccoons.

    Science.gov (United States)

    Dubey, J P; Saville, W J; Stanek, J F; Lindsay, D S; Rosenthal, B M; Oglesbee, M J; Rosypal, A C; Njoku, C J; Stich, R W; Kwok, O C; Shen, S K; Hamir, A N; Reed, S M

    2001-10-24

    Equine protozoal myeloencephalitis (EPM) is a serious neurologic disease of horses in the Americas and Sarcocystis neurona is the most common etiologic agent. The distribution of S. neurona infections follows the geographical distributions of its definitive hosts, opossums (Didelphis virginiana, Didelphis albiventris). Recently, cats and skunks were reported as experimental and armadillos as natural intermediate hosts of S. neurona. In the present report, raccoons (Procyon lotor) were identified as a natural intermediate host of S. neurona. Two laboratory-raised opossums were found to shed S. neurona-like sporocysts after ingesting tongues of naturally-infected raccoons. Interferon-gamma gene knockout (KO) mice fed raccoon-opossum-derived sporocysts developed neurologic signs. S. neurona was identified immunohistochemically in tissues of KO mice fed sporocysts and the parasite was isolated in cell cultures inoculated with infected KO mouse tissues. The DNA obtained from the tongue of a naturally-infected raccoon, brains of KO mice that had neurological signs, and from the organisms recovered in cell cultures inoculated with brains of neurologic KO mice, corresponded to that of S. neurona. Two raccoons fed mature S. neurona sarcocysts did not shed sporocysts in their feces, indicating raccoons are not likely to be its definitive host. Two raccoons fed sporocysts from opossum feces developed clinical illness and S. neurona-associated encephalomyelitis was found in raccoons killed 14 and 22 days after feeding sporocysts; schizonts and merozoites were seen in encephalitic lesions.

  14. Improving Diagnosis and Treatment of Staphylococcus aureus Infections : Experimental Studies

    NARCIS (Netherlands)

    S. van den Berg (Sanne)

    2015-01-01

    markdownabstract__Abstract__ Staphylococcus aureus is an opportunistic pathogen that causes a variety of infections, ranging from mild skin infections like furuncles and impetigo, to severe, lifethreatening infections including endocarditis, osteomyelitis and pneumonia. Invasive infections are

  15. Humoral immunity through immunoglobulin M protects mice from an experimental actinomycetoma infection by Nocardia brasiliensis.

    Science.gov (United States)

    Salinas-Carmona, Mario C; Pérez-Rivera, Isabel

    2004-10-01

    An experimental model of infection with Nocardia brasiliensis, used as an example of a facultative intracellular pathogen, was tested. N. brasiliensis was injected into the rear foot pads of BALB/c mice to establish an infection. Within 30 days, infected animals developed a chronic actinomycetoma infection. Batch cultures of N. brasiliensis were used to purify P61, P38, and P24 antigens; P61 is a catalase, and P38 is a protease with strong caseinolytic activity. Active and passive immunizations of BALB/c mice with these three purified soluble antigens were studied. Protection was demonstrated for actively immunized mice. However, immunity lasted only 30 days. Other groups of immunized mice were bled at different times, and their sera were passively transferred to naive recipients that were then infected with N. brasiliensis. Sera collected 5, 6, and 7 days after donor immunization conferred complete, long-lasting protection. The protective effect of passive immunity decreased when sera were collected 2 weeks after donor immunization. However, neither the early sera (1-, 2-, and 3-day sera) nor the later sera (30- or 45-day sera) prevented the infection. Hyperimmune sera with the highest levels of immunoglobulin G (IgG) to N. brasiliensis antigens did not protect at all. The antigens tested induced two IgM peaks. The first peak was present 3 days after immunization but was not antigen specific and did not transfer protection. The second peak was evident 7 days after immunization, was an IgM response, was antigen specific, and conferred protection. This results clearly demonstrate that IgM antibodies protect the host against a facultative intracellular bacterium.

  16. Molecular identification of Heterakis spumosa obtained from brown rats (Rattus norvegicus) in Japan and its infectivity in experimental mice.

    Science.gov (United States)

    Šnábel, Viliam; Utsuki, Daisuke; Kato, Takehiro; Sunaga, Fujiko; Ooi, Hong-Kean; Gambetta, Barbara; Taira, Kensuke

    2014-09-01

    Heterakis spumosa is a nematode of invasive rodents, mainly affiliated with Rattus spp. of Asian origin. Despite the ecological importance and cosmopolitan distribution, little information is available on the genetic characteristics and infectivity to experimental animals of this roundworm. Heterakis isolates obtained from naturally infected brown rats caught in 2007 in the city of Sagamihara, east central Honshu, Japan, and maintained by laboratory passages were subjected to mitochondrial sequence analysis and experimental infection in mice. Sequencing of the cox1 gene revealed that nucleotides of H. spumosa and previously examined Heterakis isolonche isolates from gallinaceous birds in Japan differed by 11.2-12.2% that conforms to the range expected for interspecific differences. The two H. spumosa isolates differed by a single 138T/C non-synonymous substitution in the 393-bp mt sequence. In a dendrogram, the H. spumosa samples formed a subcluster with members of the nematode superfamily Heterakoidea, H. isolonche and Ascaridia galli. In an experimental infection study, ICR, AKR, B10.BR and C57BL/6 mice strains were inoculated with 200 H. spumosa eggs/head and necropsied at 14 and 90 days post-inoculation (DPI) when the number of worms was recorded. Eggs were initially detected in faeces from 32-35 DPI in ICR, AKR and B10.BR mice and the highest mean number of eggs per gram of faeces (EPG) was 4,800 at 38 DPI, 2,200 at 58 DPI and 800 at 44 and 72 DPI in ICR, AKR and B10.BR mice, respectively. No eggs were observed in faeces of the C57BL/6 mouse strain during the experiment. A similar number of juvenile worms were isolated from all mouse strains at 14 DPI, whereas no adult worms were detected in C57BL/6 mice at 90 DPI.

  17. Avian influenza in shorebirds: experimental infection of ruddy turnstones (Arenaria interpres) with avian influenza virus

    OpenAIRE

    Hall, Jeffrey S.; Krauss, Scott; Franson, J. Christian; TeSlaa, Joshua L.; Nashold, Sean W.; Stallknecht, David E.; Webby, Richard J.; Webster, Robert G.

    2012-01-01

    Please cite this paper as: Hall et al. (2012) Avian influenza in shorebirds: experimental infection of ruddy turnstones (Arenaria interpres) with avian influenza virus. Influenza and Other Respiratory Viruses DOI: 10.1111/j.1750‐2659.2012.00358.x. Background  Low pathogenic avian influenza viruses (LPAIV) have been reported in shorebirds, especially at Delaware Bay, USA, during spring migration. However, data on patterns of virus excretion, minimal infectious doses, and clinical outcome are l...

  18. Compatibilidad entre nueve cepas de Biomphalaria glabrata de áreas endémicas y no endémicas y una cepa de Schistosoma mansoni venezolanas

    OpenAIRE

    Pino,Luz A.; Matinella,Liboria; Morales C.,Gustavo

    1999-01-01

    Se infectaron experimentalmente 9 lotes de 32 caracoles B. glabrata (de 5 a 7mm de diámetro) con miracidios de la cepa C5 de Schistosoma mansoni a razón de 5 miracidios por caracol, pertenecientes a las siguientes cepas: En el área endémica de transmisión de Esquistosomiasis mansoni a) Sector Puerta Negra, Lago Valencia, b) Cagua c) Ingenio Bolívar (Estado Aragua) d) Mariara e) Caserío El 25 f) Güigüe (Estado Carabobo). Fuera del área endémica de transmisión g) Anzoátegui (Estado Lara), h) Ch...

  19. Circulating and broncho-alveolar interleukin-6 in relation to body temperature in an experimental model of bovine Chlamydia psittaci infection.

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    Annette Prohl

    Full Text Available In rodent models of experimentally induced fever, the important role of interleukin-6 (IL-6 as a circulating endogenous pyrogen is well established. Studies employing larger animal species and real infections are scarce. Therefore, we assessed bioactive IL-6 in peripheral blood and in broncho-alveolar lavage fluid (BALF of calves after intra-bronchial inoculation with vital Chlamydia psittaci (Cp, with inactivated Cp, or with BGM cells. Only calves inoculated with vital Cp developed fever (peak at 2-3 days after challenge and significantly increased IL-6 activity. Controls inoculated with either inactivated Cp or BGM cells also expressed increased bioactive IL-6, but no fever developed. Activity of IL-6 in BALF was significantly higher compared to blood serum. This experimental model of Cp infection revealed no apparent relation between IL-6 in blood and body temperature, but did reveal a relation between IL-6 and other markers of inflammation in BALF. We conclude that a local inflammatory response in the lungs of infected calves caused fever, which developed by mechanisms including other mediators besides IL-6.

  20. Fungal infection risk groups among school children

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    Elżbieta Ejdas

    2014-08-01

    Full Text Available The aim of the study was to evaluate the relationship between ocurrence of fungi in children and living environment (city - countryside, sex, age, diet, undergone diseases therapy with antibiotics and exposure to hospital environment, and to indicate children potentially vulnerable to fungal infections. The material was consisted of swabs collected from the oral cavily, the throat and the nose of healthy children, aged 6-9 and 10-15, from both urban and rural environmens. Candida albicans, the basic aetiological factor in thc majority of mycoses recorded in humans, unquestionably prevailed in the group of the 13 speciec of yeast-like fungi and yeasts isolated. Records of C. glabrata and C. krusei increasing numbers of whose strains show resistance to basic antimycoties, as well as relatively frequent records of Trichosporon beigelii, Saccharomycopsis capsularis and Saccharomyces sp., fungi whose expansiveness and enzymatic activity have been growing, may be considered disconcerting. Vulnerability to fungal infection increases following anti-bacterial antibiotic therapy in the majority of subjects regardless season or age. This is particularly true primarily of the most stable ontocoenosis of the throat. Younger children, on the other hand, are the most vulnerable foUowing infection of the respiratory system. Fungi are likely to colonise the nose in this case. Children living in the countryside who had been ll immediately prior to the collection of the material constitute the highest risk group of the occurrence of fungi in any of the ontocoenoses studied. A greater number of positive inoculations were recorded in these children in comparison to the children from the city. It may be indicative of a more extensive spectrum of natural reservoirs of fungi and the vectors of their transmission in rural areas than those in the city, lower health hygiene and lower immunity or of a more common carriage of fungi among rural children.

  1. Effect of Experimental Coccidiosis Infections on Body Weight Gain ...

    African Journals Online (AJOL)

    Infections with E. tenella in broiler breeder males showed that body weight gains of the uninfected males were significantly greater (p< 0.05) at 5, 7 and 14 days post inoculation (dpi) than those of the infected groups. Sperm productions at 0, 5 and 7 dpi (0=day of inoculation with infected oocysts) for the uninfected controls ...

  2. Detection and localization of rabbit hepatitis e virus and antigen in systemic tissues from experimentally intraperitoneally infected rabbits.

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    Jingjing Mao

    Full Text Available Rabbit hepatitis E virus (HEV is a novel genotype of HEV, and is considered to pose a risk of zoonotic transmission. Research into the systemic distribution of rabbit HEV in rabbits during different periods of infection has rarely been reported. To better understand this virus, we infected rabbits with second-passage rabbit HEV via an intraperitoneal route. After inoculation, the infection showed two types, temporary and constant infection. The detection of HEV RNA in the feces varied with time, and serum antigen correlated with fecal HEV RNA. Viremia only appeared 72 days after inoculation. The rabbits remained antibody negative throughout the experimental period. When HEV was localized, several organs besides the liver were HEV RNA positive. Tissue antigen was observed immunohistochemically in the different cells of various organs, especially in parts of the small intestine and the characteristic rabbit gut-associated lymphoid tissue. These data provide valuable information for future research into the pathogenesis of HEV.

  3. Monitoring Activity for Recognition of Illness in Experimentally Infected Weaned Piglets Using Received Signal Strength Indication ZigBee-based Wireless Acceleration Sensor

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    Sonia Tabasum Ahmed

    2016-01-01

    Full Text Available In this experiment, we proposed and implemented a disease forecasting system using a received signal strength indication ZigBee-based wireless network with a 3-axis acceleration sensor to detect illness at an early stage by monitoring movement of experimentally infected weaned piglets. Twenty seven piglets were divided into control, Salmonella enteritidis (SE infection, and Escherichia coli (EC infection group, and their movements were monitored for five days using wireless sensor nodes on their backs. Data generated showed the 3-axis movement of piglets (X-axis: left and right direction, Y-axis: anteroposterior direction, and Z-axis: up and down direction at five different time periods. Piglets in both infected groups had lower weight gain and feed intake, as well as higher feed conversion ratios than the control group (p<0.05. Infection with SE and EC resulted in reduced body temperature of the piglets at day 2, 4, and 5 (p<0.05. The early morning X-axis movement did not differ between groups; however, the Y-axis movement was higher in the EC group (day 1 and 2, and the Z-axis movement was higher in the EC (day 1 and SE group (day 4 during different experimental periods (p<0.05. The morning X and Y-axis movement did not differ between treatment groups. However, the Z-axis movement was higher in both infected groups at day 1 and lower at day 4 compared to the control (p<0.05. The midday X-axis movement was significantly lower in both infected groups (day 4 and 5 compared to the control (p<0.05, whereas the Y-axis movement did not differ. The Z-axis movement was highest in the SE group at day 1 and 2 and lower at day 4 and 5 (p<0.05. Evening X-axis movement was highest in the control group throughout the experimental period. During day 1 and 2, the Z-axis movement was higher in both of the infected groups; whereas it was lower in the SE group during day 3 and 4 (p<0.05. During day 1 and 2, the night X-axis movement was lower and the Z

  4. Efficacy of a Gal-lectin subunit vaccine against experimental Entamoeba histolytica infection and colitis in baboons (Papio sp.).

    Science.gov (United States)

    Abd Alla, Mohamed D; Wolf, Roman; White, Gary L; Kosanke, Stanley D; Cary, David; Verweij, Jaco J; Zhang, Mie-Jie; Ravdin, Jonathan I

    2012-04-26

    To determine the efficacy of a Gal-lectin based intranasal synthetic peptide vaccine, we developed a new experimental primate model of Entamoeba histolytica intestinal infection. Release of xenic E. histolytica trophozoites (5×10(6)) into the small bowel of baboons (Papio sp.) resulted in a rapid intestinal anti-amebic antibody response and a brief infection; however, release of trophozoites directly into the cecum (5 baboons) elicited a sustained E. histolytica infection, as determined by quantitative fecal PCR, and an ulcerative, inflammatory colitis observed on colonoscopy and histopathology. In three controlled experiments, baboons received four immunizations at seven day intervals of 1600 μg of the vaccine/nostril, with Cholera toxin, 20 μg/nostril as adjuvant; vaccinated (n=6) and control baboons (n=6) baboons were then challenged via colonoscopy with xenic trophozoites (5×10(6)). During 90 days of follow up, 250 of 415 (60.24%) fecal samples in control baboons had a (+) PCR for E. histolytica, compared to only 36 of 423 (8.51%) samples from vaccinated baboons (P<0.001). All 6 vaccinated baboons were free of infection by the 51st day after challenge, 5 of 6 controls positive had (+) fecal PCRs for up to 126 days post-challenge (P=0.019). Inflammatory colitis developed in 4 of 6 control baboons post-challenge, with invasive E. histolytica trophozoites present in 2 of the 4 on histopathology. There was no evidence of inflammatory colitis or parasite invasion in any of the vaccinated baboons; there was a strong inverse correlation between positive ELISA OD value indicating the presence of intestinal anti-peptide IgA antibodies and baboons having a positive fecal PCR CT value, P<0.001. In conclusion, we developed a novel primate model of E. histolytica intestinal infection and demonstrated that a Gal-lectin-based intranasal synthetic peptide vaccine was highly efficacious in preventing experimental E. histolytica infection and colitis in baboons. Copyright

  5. Experimental Pseudomonas anguilliseptica infection in turbot Psetta maxima (L.: a histopathological and immunohistochemical study

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    JL Romalde

    2009-06-01

    Full Text Available Experimental infection with Pseudomonas anguilliseptica was performed both by intraperitoneal (i.p. and bath route on juvenile turbot (Psetta maxima in order to evaluate the pathology induced. Turbot was found to be sensitive to i.p. challenge (1.7x106 CFU/fish but no to bath exposure. The i.p. challenge induced septicaemic infection and mortality. Externally, moribund fish showed distended abdomen and pale areas at day 9. The gross pathological internal signs present were abundant ascitic fluid in the peritoneal cavity, pale and enlarged spleen, pale and friable liver, and congestive and dilated gut with yellowish exudates. On histopathological examination, bacterial invasion was common in all the tissues studied but the most prominent pathological changes were observed in gut, spleen and kidney after 7 day with features of necrosis. The immunohistochemical findings support the widespread localization of the bacteria after the i.p. injection since the P. anguilliseptica was detected in spleen from day 1 post injection, in liver, kidney and gut from day 4, in muscle from day 7 and in brain from day 9. The difficulties in infecting healthy fish by bath challenge can be explained by the opportunistic nature of this pathogen.

  6. Experimental infection with the Toxoplasma gondii ME-49 strain in the Brazilian BR-1 mini pig is a suitable animal model for human toxoplasmosis

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    Farlen José Bebber Miranda

    2015-02-01

    Full Text Available Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain or orally infected with cysts (ME-49 strain. Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs' peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis.

  7. Experimental Infection of Rhodnius prolixus (Hemiptera, Triatominae with Mycobacterium leprae Indicates Potential for Leprosy Transmission.

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    Arthur da Silva Neumann

    Full Text Available Leprosy is a chronic dermato-neurological disease caused by infection with Mycobacterium leprae. In 2013 almost 200,000 new cases of leprosy were detected around the world. Since the first symptoms take from years to decades to appear, the total number of asymptomatic patients is impossible to predict. Although leprosy is one of the oldest records of human disease, the mechanisms involved with its transmission and epidemiology are still not completely understood. In the present work, we experimentally investigated the hypothesis that the mosquitoes Aedes aegypti and Culex quinquefasciatus and the hemiptera Rhodnius prolixus act as leprosy vectors. By means of real-time PCR quantification of M. leprae 16SrRNA, we found that M. leprae remained viable inside the digestive tract of Rhodnius prolixus for 20 days after oral infection. In contrast, in the gut of both mosquito species tested, we were not able to detect M. leprae RNA after a similar period of time. Inside the kissing bug Rhodnius prolixus digestive tract, M. leprae was initially restricted to the anterior midgut, but gradually moved towards the hindgut, in a time course reminiscent of the life cycle of Trypanosoma cruzi, a well-known pathogen transmitted by this insect. The maintenance of M. leprae infectivity inside the digestive tract of this kissing bug is further supported by successful mice footpad inoculation with feces collected 20 days after infection. We conclude that Rhodnius prolixus defecate infective M. leprae, justifying the evaluation of the presence of M. leprae among sylvatic and domestic kissing bugs in countries endemic for leprosy.

  8. Experimental Infection of Rhodnius prolixus (Hemiptera, Triatominae) with Mycobacterium leprae Indicates Potential for Leprosy Transmission.

    Science.gov (United States)

    Neumann, Arthur da Silva; Dias, Felipe de Almeida; Ferreira, Jéssica da Silva; Fontes, Amanda Nogueira Brum; Rosa, Patricia Sammarco; Macedo, Rafael Enrique; Oliveira, José Henrique; Teixeira, Raquel Lima de Figueiredo; Pessolani, Maria Cristina Vidal; Moraes, Milton Ozório; Suffys, Philip Noel; Oliveira, Pedro L; Sorgine, Marcos Henrique Ferreira; Lara, Flavio Alves

    2016-01-01

    Leprosy is a chronic dermato-neurological disease caused by infection with Mycobacterium leprae. In 2013 almost 200,000 new cases of leprosy were detected around the world. Since the first symptoms take from years to decades to appear, the total number of asymptomatic patients is impossible to predict. Although leprosy is one of the oldest records of human disease, the mechanisms involved with its transmission and epidemiology are still not completely understood. In the present work, we experimentally investigated the hypothesis that the mosquitoes Aedes aegypti and Culex quinquefasciatus and the hemiptera Rhodnius prolixus act as leprosy vectors. By means of real-time PCR quantification of M. leprae 16SrRNA, we found that M. leprae remained viable inside the digestive tract of Rhodnius prolixus for 20 days after oral infection. In contrast, in the gut of both mosquito species tested, we were not able to detect M. leprae RNA after a similar period of time. Inside the kissing bug Rhodnius prolixus digestive tract, M. leprae was initially restricted to the anterior midgut, but gradually moved towards the hindgut, in a time course reminiscent of the life cycle of Trypanosoma cruzi, a well-known pathogen transmitted by this insect. The maintenance of M. leprae infectivity inside the digestive tract of this kissing bug is further supported by successful mice footpad inoculation with feces collected 20 days after infection. We conclude that Rhodnius prolixus defecate infective M. leprae, justifying the evaluation of the presence of M. leprae among sylvatic and domestic kissing bugs in countries endemic for leprosy.

  9. Detection of arboviruses and other micro-organisms in experimentally infected mosquitoes using massively parallel sequencing.

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    Sonja Hall-Mendelin

    Full Text Available Human disease incidence attributed to arbovirus infection is increasing throughout the world, with effective control interventions limited by issues of sustainability, insecticide resistance and the lack of effective vaccines. Several promising control strategies are currently under development, such as the release of mosquitoes trans-infected with virus-blocking Wolbachia bacteria. Implementation of any control program is dependent on effective virus surveillance and a thorough understanding of virus-vector interactions. Massively parallel sequencing has enormous potential for providing comprehensive genomic information that can be used to assess many aspects of arbovirus ecology, as well as to evaluate novel control strategies. To demonstrate proof-of-principle, we analyzed Aedes aegypti or Aedes albopictus experimentally infected with dengue, yellow fever or chikungunya viruses. Random amplification was used to prepare sufficient template for sequencing on the Personal Genome Machine. Viral sequences were present in all infected mosquitoes. In addition, in most cases, we were also able to identify the mosquito species and mosquito micro-organisms, including the bacterial endosymbiont Wolbachia. Importantly, naturally occurring Wolbachia strains could be differentiated from strains that had been trans-infected into the mosquito. The method allowed us to assemble near full-length viral genomes and detect other micro-organisms without prior sequence knowledge, in a single reaction. This is a step toward the application of massively parallel sequencing as an arbovirus surveillance tool. It has the potential to provide insight into virus transmission dynamics, and has applicability to the post-release monitoring of Wolbachia in mosquito populations.

  10. Deletion of the Candida glabrata ERG3 and ERG11 genes: effect on cell viability, cell growth, sterol composition, and antifungal susceptibility.

    Science.gov (United States)

    Geber, A; Hitchcock, C A; Swartz, J E; Pullen, F S; Marsden, K E; Kwon-Chung, K J; Bennett, J E

    1995-01-01

    We have cloned and sequenced the structural genes encoding the delta 5,6 sterol desaturase (ERG3 gene) and the 14 alpha-methyl sterol demethylase (ERG11 gene) from Candida glabrata L5 (leu2). Single and double mutants of these genes were created by gene deletion. The phenotypes of these mutants, including sterol profiles, aerobic viabilities, antifungal susceptibilities, and generation times, were studied. Strain L5D (erg3 delta::LEU2) accumulated mainly ergosta-7,22-dien-3 beta-ol, was aerobically viable, and remained susceptible to antifungal agents but had a slower generation time than its parent strain. L5LUD (LEU2 erg11 delta::URA3) strains required medium supplemented with ergosterol and an anaerobic environment for growth. A spontaneous aerobically viable mutant, L5LUD40R (LEU erg11 delta::URA3), obtained from L5LUD (LEU2 erg11 delta::URA3), was found to accumulate lanosterol and obtusifoliol, was resistant to azole antifungal agents, demonstrated some increase in resistance to amphotericin B, and exhibited a 1.86-fold increase in generation time in comparison with L5 (leu2). The double-deletion mutant L5DUD61 (erg3 delta::LEU2 erg11 delta::URA3) was aerobically viable, produced mainly 14 alpha-methyl fecosterol, and had the same antifungal susceptibility pattern as L5LUD40R (LEU2 erg11 delta::URA3), and its generation time was threefold greater than that of L5 (leu2). Northern (RNA) analysis revealed that the single-deletion mutants had a marked increase in message for the undeleted ERG3 and ERG11 genes. These results indicate that differences in antifungal susceptibilities and the restoration of aerobic viability exist between the C. glabrata ergosterol mutants created in this study and those sterol mutants with similar genetic lesions previously reported for Saccharomyces cerevisiae. PMID:8593007

  11. Efficacy of a 60Co irradiated vaccine for experimentally infected calves with dictyocaulus viviparus

    International Nuclear Information System (INIS)

    Zurita, Edgar; Paredes, Julio; Fernandez, Ardey

    1991-01-01

    Dictyocaulus viviparus larvae in non-infected stage (L.1) were cultured in vitro to their infective stage (L.3) and were irradiated with 40 Krad from 6 0 C o and used as a vaccine. The oral experimental vaccine dose was 1000 L.3/animal. Three groups were formed with 8 calves in each one: group No. 1 and group No. 2 were vaccinated at 10 weeks of age. Four weeks later group No. 1 was infected with non-irradiated 60 L.3/Kg. of animal weight 'challenge dose'. After four weeks of post-vaccination group No. 2 was revaccinated and 4 weeks later it was infected with the 'challenge dose' as the previous group. Eight calves constituted group No. 3, four in each group; they were infected with only the challenge dose respectively. Information data on respiratory and cardiac frequency, temperature, weight, and L.1 count/g. of feces were obtained from each animal three times a week. After five weeks of post-challenge all animals were slaughtered to observe anatomopathologic lessions in the heart and lungs; the number of adult Dictyocaulus viviparus present in the respiratory tract were search 't'. Student test was used for the statistical analysis. The weight increment difference between animals of group No. 1 related control group was 7Kg.; and 11.25 Kg/animal in group No. 2 respectively. The percentage of protection confered by the vaccine to the subjects in group No. 1 relative to the respectivecontrol group, was 83.2; that for the group No. 2 was 88.5 per cent. Post-morten examination revealed severe anatomopathologic lessions in the control groups; only few lessions were observed in group No. 1 and practically none in group No. 2

  12. Spectrum of Opportunistic Fungal Infections in HIV/AIDS Patients in Tertiary Care Hospital in India

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    Ravinder Kaur

    2016-01-01

    Full Text Available HIV related opportunistic fungal infections (OFIs continue to cause morbidity and mortality in HIV infected patients. The objective for this prospective study is to elucidate the prevalence and spectrum of common OFIs in HIV/AIDS patients in north India. Relevant clinical samples were collected from symptomatic HIV positive patients (n=280 of all age groups and both sexes and subjected to direct microscopy and fungal culture. Identification as well as speciation of the fungal isolates was done as per the standard recommended methods. CD4+T cell counts were determined by flow cytometry using Fluorescent Activated Cell Sorter Count system. 215 fungal isolates were isolated with the isolation rate of 41.1%. Candida species (86.5% were the commonest followed by Aspergillus (6.5%, Cryptococcus (3.3%, Penicillium (1.9%, and Alternaria and Rhodotorula spp. (0.9% each. Among Candida species, Candida albicans (75.8% was the most prevalent species followed by C. tropicalis (9.7%, C. krusei (6.4%, C. glabrata (4.3%, C. parapsilosis (2.7%, and C. kefyr (1.1%. Study demonstrates that the oropharyngeal candidiasis is the commonest among different OFIs and would help to increase the awareness of clinicians in diagnosis and early treatment of these infections helping in the proper management of the patients especially in resource limited countries like ours.

  13. Cis and trans acting factors involved in human cytomegalovirus experimental and natural latent infection of CD14 (+ monocytes and CD34 (+ cells.

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    Cyprian C Rossetto

    Full Text Available The parameters involved in human cytomegalovirus (HCMV latent infection in CD14 (+ and CD34 (+ cells remain poorly identified. Using next generation sequencing we deduced the transcriptome of HCMV latently infected CD14 (+ and CD34 (+ cells in experimental as well as natural latency settings. The gene expression profile from natural infection in HCMV seropositive donors closely matched experimental latency models, and included two long non-coding RNAs (lncRNAs, RNA4.9 and RNA2.7 as well as the mRNAs encoding replication factors UL84 and UL44. Chromatin immunoprecipitation assays on experimentally infected CD14 (+ monocytes followed by next generation sequencing (ChIP-Seq were employed to demonstrate both UL84 and UL44 proteins interacted with the latent viral genome and overlapped at 5 of the 8 loci identified. RNA4.9 interacts with components of the polycomb repression complex (PRC as well as with the MIE promoter region where the enrichment of the repressive H3K27me3 mark suggests that this lncRNA represses transcription. Formaldehyde Assisted Isolation of Regulatory Elements (FAIRE, which identifies nucleosome-depleted viral DNA, was used to confirm that latent mRNAs were associated with actively transcribed, FAIRE analysis also showed that the terminal repeat (TR region of the latent viral genome is depleted of nucleosomes suggesting that this region may contain an element mediating viral genome maintenance. ChIP assays show that the viral TR region interacts with factors associated with the pre replication complex and a plasmid subclone containing the HCMV TR element persisted in latently infected CD14 (+ monocytes, strongly suggesting that the TR region mediates viral chromosome maintenance.

  14. Efficacy of the amino-acetonitrile derivative, monepantel, against experimental and natural adult stage gastro-intestinal nematode infections in sheep.

    Science.gov (United States)

    Sager, Heinz; Hosking, Barry; Bapst, Béatrice; Stein, Philip; Vanhoff, Kathleen; Kaminsky, Ronald

    2009-01-22

    Multiple drug resistance by nematodes, against anthelmintics has become an important economic problem in sheep farming worldwide. Here we describe the efficacy of monepantel, a developmental molecule from the recently discovered anthelmintic class, the amino-acetonitrile derivatives (AADs). Efficacy was tested against adult stage gastro-intestinal nematodes (GINs) in experimentally and naturally infected sheep at a dose of 2.5mg/kg body weight when administered as an oral solution. Some of the isolates used in experimental infection studies were known to be resistant to the benzimidazoles or levamisole anthelmintics; strains resistant to the macrocyclic lactones were not available for these tests. Worm count-based efficacies of >98% were determined in these studies. As an exception, Oesophagostomum venulosum was only reduced by 88% in one study, albeit with a low worm burden in the untreated controls (geometric mean 15.4 worms). Similar efficacies for monepantel were also confirmed in naturally infected sheep. While the efficacy against most species was >99%, the least susceptible species was identified as Nematodirus spathiger, and although efficacy was 92.4% in one study it was generally >99%. Several animals were infected with Trichuris ovis, which was not eliminated after the treatment. Monepantel demonstrated high activity against a broad range of the important GINs of sheep, which makes this molecule an interesting candidate for use in this species, particularly in regions with problems of anthelmintic resistance. Monepantel was well tolerated by the treated sheep, with no treatment related adverse events documented.

  15. Ivermectin: activity against larval Strongylus vulgaris and adult Trichostrongylus axei in experimental infections in ponies.

    Science.gov (United States)

    Lyons, E T; Drudge, J H; Tolliver, S C

    1982-08-01

    Activity of ivermectin, administered IM at the dosage rate of 200 micrograms/kg of body weight, was evaluated in controlled tests against migrating larvae of Strongylus vulgaris and adult Trichostrongylus axei in experimental infections in 6 ponies raised worm-free. Ponies were given 2,190 or 2,400 infective 3rd-stage larvae of S vulgaris at 7 days before treatment and 22,000 or 22,750 infective 3rd-stage larvae of T axei at 42 or 45 days before treatment. Three ponies were given ivermectin plus vehicle, and 3 ponies were given the vehicle only; the ponies were euthanatized 7 or 9 days after treatment. At necropsy, 4th-stage S vulgaris larvae were not recovered from visceral arteries of the 3 ivermectin plus vehicle-treated ponies, but 21 to 40 larvae were recovered from each of the 3 vehicle-treated ponies. Also at necropsy, adult T axei (140 specimens) were recovered from only 1 ot the 3 ivermectin plus vehicle-treated ponies, but 4,610 to 6,410 specimens were found in each of the 3 vehicle-treated ponies. Toxicosis was not observed after treatment.

  16. Experimental Cowpox Virus (CPXV) Infections of Bank Voles: Exceptional Clinical Resistance and Variable Reservoir Competence.

    Science.gov (United States)

    Franke, Annika; Ulrich, Rainer G; Weber, Saskia; Osterrieder, Nikolaus; Keller, Markus; Hoffmann, Donata; Beer, Martin

    2017-12-19

    Cowpox virus (CPXV) is a zoonotic virus and endemic in wild rodent populations in Eurasia. Serological surveys in Europe have reported high prevalence in different vole and mouse species. Here, we report on experimental CPXV infections of bank voles ( Myodes glareolus ) from different evolutionary lineages with a spectrum of CPXV strains. All bank voles, independently of lineage, sex and age, were resistant to clinical signs following CPXV inoculation, and no virus shedding was detected in nasal or buccal swabs. In-contact control animals became only rarely infected. However, depending on the CPXV strain used, inoculated animals seroconverted and viral DNA could be detected preferentially in the upper respiratory tract. The highest antibody titers and virus DNA loads in the lungs were detected after inoculation with two strains from Britain and Finland. We conclude from our experiments that the role of bank voles as an efficient and exclusive CPXV reservoir seems questionable, and that CPXV may be maintained in most regions by other hosts, including other vole species. Further investigations are needed to identify factors that allow and modulate CPXV maintenance in bank voles and other potential reservoirs, which may also influence spill-over infections to accidental hosts.

  17. Dynamics of Mixed- Candida Species Biofilms in Response to Antifungals.

    Science.gov (United States)

    Vipulanandan, G; Herrera, M; Wiederhold, N P; Li, X; Mintz, J; Wickes, B L; Kadosh, D

    2018-01-01

    Oral infections caused by Candida species, the most commonly isolated human fungal pathogen, are frequently associated with biofilms. Although Candida albicans is the predominant organism found in patients with oral thrush, a biofilm infection, there is an increasing incidence of oral colonization and infections caused by non- albicans Candida species, including C. glabrata, C. dubliniensis, and C. tropicalis, which are frequently more resistant to antifungal treatment. While single-species Candida biofilms have been well studied, considerably less is known about the dynamics of mixed- Candida species biofilms and how these dynamics are altered by antifungal treatment. To address these questions, we developed a quantitative polymerase chain reaction-based approach to determine the precise species composition of mixed- Candida species biofilms formed by clinical isolates and laboratory strains in the presence and absence of clinically relevant concentrations of 3 commonly used antifungals: fluconazole, caspofungin, and amphotericin B. In monospecies biofilms, fluconazole exposure favored growth of C. glabrata and C. tropicalis, while caspofungin generally favored significant growth of all species to a varying degree. Fluconazole was not effective against preformed mixed- Candida species biofilms while amphotericin B was potent. As a general trend, in mixed- Candida species biofilms, C. albicans lost dominance in the presence of antifungals. Interestingly, presence in mixed versus monospecies biofilms reduced susceptibility to amphotericin B for C. tropicalis and C. glabrata. Overall, our data suggest that antifungal treatment favors the growth of specific non- albicans Candida species in mixed- Candida species biofilms.

  18. Species distribution & antifungal susceptibility pattern of oropharyngeal Candida isolates from human immunodeficiency virus infected individuals

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    Partha Pratim Das

    2016-01-01

    Results: From the 59 culture positive HIV seropositive cases, 61 Candida isolates were recovered; Candidaalbicans (n=47, 77.0%, C. dubliniensis (n=9, 14.7%, C. parapsilosis (n=2, 3.2%, C. glabrata (n=2, 3.2%, and C. famata (n=1, 1.6%. Candida colonization in HIV-seropositive individuals was significantly higher than that of HIV-seronegative (control group. Antifungal susceptibility testing revealed (n=6, 9.3% C. albicans isolates resistant to voriconazole and fluconazole by disk-diffusion method whereas no resistance was seen by Fungitest method. Interpretation & conclusions: C. albicans was the commonest Candida species infecting or colonizing HIV seropositive individuals. Oropharyngeal Candida isolates had high level susceptibility to all the major antifungals commonly in use. Increased level of immunosuppression in HIV-seropositives and drug resistance of non-albicans Candida species makes identification and susceptibility testing of Candida species necessary in different geographical areas of the country.

  19. IL-6 is Upregulated in Late-Stage Disease in Monkeys Experimentally Infected with Trypanosoma brucei rhodesiense

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    Dawn Nyawira Maranga

    2013-01-01

    Full Text Available The management of human African trypanosomiasis (HAT is constrained by lack of simple-to-use diagnostic, staging, and treatment tools. The search for novel biomarkers is, therefore, essential in the fight against HAT. The current study aimed at investigating the potential of IL-6 as an adjunct parameter for HAT stage determination in vervet monkey model. Four adult vervet monkeys (Chlorocebus aethiops were experimentally infected with Trypanosoma brucei rhodesiense and treated subcuratively at 28 days after infection (dpi to induce late stage disease. Three noninfected monkeys formed the control group. Cerebrospinal fluid (CSF and blood samples were obtained at weekly intervals and assessed for various biological parameters. A typical HAT-like infection was observed. The late stage was characterized by significant (P<0.05 elevation of CSF IL-6, white blood cell count, and total protein starting 35 dpi with peak levels of these parameters coinciding with relapse parasitaemia. Brain immunohistochemical staining revealed an increase in brain glial fibrillary acidic protein expression indicative of reactive astrogliosis in infected animals which were euthanized in late-stage disease. The elevation of IL-6 in CSF which accompanied other HAT biomarkers indicates onset of parasite neuroinvasion and show potential for use as an adjunct late-stage disease biomarker in the Rhodesian sleeping sickness.

  20. Avian influenza in shorebirds: experimental infection of ruddy turnstones (Arenaria interpres) with avian influenza virus

    Science.gov (United States)

    Hall, Jeffrey S.; Krauss, Scott; Franson, J. Christian; TeSlaa, Joshua L.; Nashold, Sean W.; Stallknecht, David E.; Webby, Richard J.; Webster, Robert G.

    2013-01-01

    Background: Low pathogenic avian influenza viruses (LPAIV) have been reported in shorebirds, especially at Delaware Bay, USA, during spring migration. However, data on patterns of virus excretion, minimal infectious doses, and clinical outcome are lacking. The ruddy turnstone (Arenaria interpres) is the shorebird species with the highest prevalence of influenza virus at Delaware Bay. Objectives: The primary objective of this study was to experimentally assess the patterns of influenza virus excretion, minimal infectious doses, and clinical outcome in ruddy turnstones. Methods: We experimentally challenged ruddy turnstones using a common LPAIV shorebird isolate, an LPAIV waterfowl isolate, or a highly pathogenic H5N1 avian influenza virus. Cloacal and oral swabs and sera were analyzed from each bird. Results: Most ruddy turnstones had pre-existing antibodies to avian influenza virus, and many were infected at the time of capture. The infectious doses for each challenge virus were similar (103·6–104·16 EID50), regardless of exposure history. All infected birds excreted similar amounts of virus and showed no clinical signs of disease or mortality. Influenza A-specific antibodies remained detectable for at least 2 months after inoculation. Conclusions: These results provide a reference for interpretation of surveillance data, modeling, and predicting the risks of avian influenza transmission and movement in these important hosts.

  1. Experimental infection of octopus vulgaris (Cuvier, 1797) with Photobacterium damsela subsp. piscicida. Immunohistochemical tracking of antigen and tissue responses.

    Science.gov (United States)

    Bakopoulos, Vasileios; White, Daniella; Valsamidis, Michail-Aggelos; Vasilaki, Feli

    2017-03-01

    Adult common octopus individuals were intramuscularly infected with Photobacterium damsela subsp. piscicida in order to investigate if this species is sensitive to this common and important fish pathogen. The fate of the bacterial antigens and the tissue responses of Octopus vulgaris were studied employing immunohistochemical techniques. Strong reaction at the site of injection was evident from day 2 post-infection that continued until day 14. Great numbers of hemocytes that were attracted at the site of infection were involved in phagocytosis of bacteria. Very early in the infection, a transition of cells to fibroblasts and an effort to isolate the infection was observed. During the course of the study, very large necrotic cells were seen at the site of infection, whereas during the later stages hemocytes with phagocytosed bacteria were observed in well-defined pockets inside the muscle tissue. None of the internal organs tested for the presence of the bacterium were positive with the exception of the digestive gland where antigen staining was observed which was not associated with hemocyte infiltration. The high doses of bacterial cells used in this experimental infection and the lack of disease signs from Octopus vulgaris suggest that, under normal conditions, octopus is resistant to Photobacterium damsela subsp. piscicida. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Experimental intraocular infection of exotic cockerels with field strain of velogenic Newcastle disease virus in Nigeria

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    Samaila Jonathan Badau

    2015-12-01

    Full Text Available Experimental intraocular (conjunctival infection of exotic cockerels with a new field strain of viscerotropic velogenic Newcastle Disease Virus (NDV was conducted to explore the concurrence of some pathological changes with humoral immune responses. After the NDV infection of 4-week-old cockerels, pathologic changes and antibody responses were observed. The clinical signs observed after the artificial inoculation included inappetence, depression, diarrhea, dyspnea, wing and leg paralysis, torticollis and weight loss. Morbidity due to the NDV was 100%, but mortality was 80% by day 18-21 post-infection. Early hyperthermia followed by terminal hypothermia, decreased packed cell volume (PCV, and 231.4 folds peak-antibody response were observed. Necrotic and/or inflammatory lesions were present in the proventriculus, intestine, liver, spleen, kidney and brain. Neurologic and digestive tract perturbations occurred in 10% and 85% of cases, respectively. The disease consistently caused stunted growth, decreased PCV, and necro-inflammatroy lesions concurrent with antibody response, suggesting probable involvement of immune-mediated mechanisms and cell membrane desialylation by viral neuraminidase in the pathogenesis.

  3. Neither hippurate-negative Brachyspira pilosicoli nor Brachyspira pilosicoli type strain caused diarrhoea in early-weaned pigs by experimental infection

    DEFF Research Database (Denmark)

    Fossi, M.; Ahlsten, K.; Pohjanvirta, T.

    2005-01-01

    Fossi M, Ahlsten K, Pohjanvirta T, Anttila M, Kokkonen T, Jensen TK, Boye M, Sukura A, Pelkola K, Pelkonen S: Neither hippurate-negative Brachyspira pilosicoli nor Brachyspira pilosicoli type strain caused diarrhoea in early-weaned pigs by experimental infection. Acta vet. scand. 2005, 46, 257...

  4. Antifungal Activity of 14-Helical β-Peptides against Planktonic Cells and Biofilms of Candida Species

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    Namrata Raman

    2015-08-01

    Full Text Available Candida albicans is the most prevalent cause of fungal infections and treatment is further complicated by the formation of drug resistant biofilms, often on the surfaces of implanted medical devices. In recent years, the incidence of fungal infections by other pathogenic Candida species such as C. glabrata, C. parapsilosis and C. tropicalis has increased. Amphiphilic, helical β-peptide structural mimetics of natural antimicrobial α-peptides have been shown to exhibit specific planktonic antifungal and anti-biofilm formation activity against C. albicans in vitro. Here, we demonstrate that β-peptides are also active against clinically isolated and drug resistant strains of C. albicans and against other opportunistic Candida spp. Different Candida species were susceptible to β-peptides to varying degrees, with C. tropicalis being the most and C. glabrata being the least susceptible. β-peptide hydrophobicity directly correlated with antifungal activity against all the Candida clinical strains and species tested. While β-peptides were largely ineffective at disrupting existing Candida biofilms, hydrophobic β-peptides were able to prevent the formation of C. albicans, C. glabrata, C. parapsilosis and C. tropicalis biofilms. The broad-spectrum antifungal activity of β-peptides against planktonic cells and in preventing biofilm formation suggests the promise of this class of molecules as therapeutics.

  5. Changes in blood sugar levels of rats experimentally infected with Trypanosoma brucei and treated with imidocarb dipropionate and diminazene aceturate

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    Nwoha Rosemary Ijeoma Ogechi

    2016-01-01

    Full Text Available Objective: To determine the effect of Trypanosoma brucei (T. brucei on blood sugar level of infected rats. Methods: The experiment was done with 42 albino rats grouped into 3 groups of 14 members each. Group A was uninfected (control group, Group B was infected with T. brucei and treated with diminazene aceturate, and Group C was infected with T. brucei and treated with imidocarb dipropionate. Blood samples were collected from the media canthus of the experimental rats on Days 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 for the assessment of change in blood sugar levels. The blood sugar levels were determined with a glucometer (Accu-chek active serial No. GN: 10023338. Results: By 4 to 5 days post infection, there was a significant increase (P 0.05 was observed in the groups when compared with the control group till Day 12 of the experiment. Conclusions: T. brucei caused a significant increase in blood sugar of infected rats.

  6. Increased pulmonary secretion of tumor necrosis factor-alpha in calves experimentally infected with bovine respiratory syncytial virus

    DEFF Research Database (Denmark)

    Rontved, C. M.; Tjørnehøj, Kirsten; Viuff, B.

    2000-01-01

    , of which 23 were experimentally infected with BRSV and five were given a mock inoculum. The presence of the cytokine tumor necrosis factor alpha (TNF-alpha) in the BAL fluids was detected and quantified by a capture ELISA. TNF-alpha was detected in 21 of the infected animals. The amount of TNF-alpha...... in the BAL fluid of calves killed post inoculation day (PID) 2 and 4 was at the same very low level as in the uninfected control animals. Large amounts of TNF-alpha were detected on PID 6, maximum levels of TNF-alpha were reached on PID 7, and smaller amounts of TNF-alpha were seen on PID 8. The high levels...... of TNF-alpha appeared on the days where severe lung lesions and clinical signs were obvious and the amounts of BRSV-antigen were at their greatest. Although Pasteurellaceae were isolated from some of the BRSV-infected calves, calves treated with antibiotics before and through the whole period...

  7. Efficacy of sulfonamides and Baycox(®) against Isospora suis in experimental infections of suckling piglets.

    Science.gov (United States)

    Joachim, Anja; Mundt, Hans-Christian

    2011-12-01

    Sulfonamide treatment of piglets against neonatal coccidiosis has frequently been suggested in the literature. In order to evaluate the efficacy of sulfonamides against experimental Isospora suis infections in suckling piglets (oral infection with 1,500 sporulated oocysts of I. suis per piglet on the fourth day of life), two trials were conducted. In trial I, oral sulfadimidine (group Sulfa-Oral) was applied in doses of 100 mg/kg of body weight (BW) 1 day before infection and 75 mg/kg BW daily for the following 5 days, and sulfamethoxypyrimidine (SMP) was applied parenterally in daily doses of 75 mg/kg BW for the same time period. In trial II, SMP was applied parenterally in doses of 75 mg/kg BW (a) from the day of infection daily for 7 days (SMP-Standard), (b) for 2 days starting on the day of infection (SMP-Early), (c) for 3 days starting 2 days post-infection (d.p.i.; SMP-Middle), (d) for 2 days starting 5 d.p.i. (SMP-Late), and (e) every other day from the day of infection until 6 d.p.i. (SMP-Alternating), as well as (f) orally in doses of 75 mg/kg BW from the day of infection for 7 days (SMP-Oral). The sulfonamide-treated groups were compared to a toltrazuril-treated group (single oral treatment with Baycox® 5% suspension, 20 mg/kg BW 2 d.p.i.) and to a water-treated Control group. Each group consisted of seven to nine piglets. The parameters evaluated were oocyst excretion and fecal consistency/diarrhea from 4 to 15 d.p.i. Sulfa-Oral, SMP-Early, and SMP-Late had no significant effect in reduction of oocyst excretion and diarrhea, whereas treatment for 3-7 days with SMP reduced both parasite shedding and diarrhea significantly. Oral treatment with SMP was comparable to parenteral application. Baycox® in a single application had the most pronounced effect and completely suppressed oocyst excretion and diarrhea during the examination period. It could be shown that repeated application of sulfonamides, provided that the appropriate time period after infection

  8. An immunohistochemical study of Flexibacter psychrophilus infection in experimentally and naturally infected rainbow trout (Oncorhynchus mykiss) fry

    DEFF Research Database (Denmark)

    Evensen, O.; Lorenzen, Ellen

    1996-01-01

    An immunohistochemical method is described for the detection of Flexibacter psychrophilus in formalin-fixed, parafiin-wax-embedded fry of rainbow trout. Rabbit antiserum as well as rainbow trout hyperimmune serum were used in the study. The distribution and tissue localization of the bacterium wa...... are typically found during the chronic stage of the disease....... and experimentally infected fry showed that there was a localization of bacteria in the monocyte-macrophage system, in skin lesions, and in the retina and the choroid gland of the eye. The dermal changes included superficial or deep ulcers extending to the subcutaneous tissue or the musculature accompanied...... by inflammatory cell infiltrates in which polymorphonuclear inflammatory cells were shown to contain the bacterium in the cytoplasm by immunostaining. The eye changes were likewise a common finding in chronic cases with severe inflammatory changes in the retina and with numerous bacteria in inflammatory (mainly...

  9. Corneal Opacity in Domestic Ducks Experimentally Infected With H5N1 Highly Pathogenic Avian Influenza Virus.

    Science.gov (United States)

    Yamamoto, Y; Nakamura, K; Yamada, M; Mase, M

    2016-01-01

    Domestic ducks can be a key factor in the regional spread of H5N1 highly pathogenic avian influenza (HPAI) virus in Asia. The authors performed experimental infections to examine the relationship between corneal opacity and H5N1 HPAI virus infection in domestic ducks (Anas platyrhyncha var domestica). A total of 99 domestic ducks, including 3 control birds, were used in the study. In experiment 1, when domestic ducks were inoculated intranasally with 2 H5N1 HPAI viruses, corneal opacity appeared more frequently than neurologic signs and mortality. Corneal ulceration and exophthalmos were rare findings. Histopathologic examinations of the eyes of domestic ducks in experiment 2 revealed that corneal opacity was due to the loss of corneal endothelial cells and subsequent keratitis with edema. Influenza viral antigen was detected in corneal endothelial cells and some other ocular cells by immunohistochemistry. Results suggest that corneal opacity is a characteristic and frequent finding in domestic ducks infected with the H5N1 HPAI virus. Confirming this ocular change may improve the detection rate of infected domestic ducks in the field. © The Author(s) 2015.

  10. Detection of H5N1 high-pathogenicity avian influenza virus in meat and tracheal samples from experimentally infected chickens.

    Science.gov (United States)

    Das, Amaresh; Spackman, Erica; Thomas, Colleen; Swayne, David E; Suarez, David L

    2008-03-01

    The Asian H5N1 highly pathogenic avian influenza (HPAI) virus causes a systemic disease with high mortality of poultry and is potentially zoonotic. In both chickens and ducks, the virus has been demonstrated to replicate in both cardiac and skeletal muscle cells. Experimentally, H5N1 HPAI virus has been transmitted to chickens through the consumption of raw infected meat. In this study, we investigated virus replication in cardiac and skeletal muscle and in the trachea of chickens after experimental intranasal inoculation with the H5N1 HPAI virus. The virus was detected in tissues by real-time reverse transcription-polymerase chain reaction (RRT-PCR) and virus isolation, and in the trachea by RRT-PCR and a commercial avian influenza (AI) viral antigen detection test. A modified RNA extraction protocol was developed for rapid detection of the virus in tissues by RRT-PCR. The H5N1 HPAI virus was sporadically detected in meat and the tracheas of infected birds without any clinical sign of disease as early as 6 hr postinfection (PI), and was detected in all samples tested at 24 hr PI and later. No differences in sensitivity were seen between virus isolation and RRT-PCR in meat samples. The AI viral antigen detection test on tracheal swabs was a useful method for identifying infected chickens when they were sick or dead, but was less sensitive in detecting infected birds when they were preclinical. This study provides data indicating that preslaughter tracheal swab testing can identify birds infected with HPAI among the daily mortality and prevent infected flocks from being sent to processing plants. In addition, the modified RNA extraction and RRT-PCR test on meat samples provide a rapid and sensitive method of identifying HPAI virus in illegal contraband or domestic meat samples.

  11. Experimental Model of Tuberculosis in the Domestic Goat after Endobronchial Infection with Mycobacterium caprae ▿

    Science.gov (United States)

    Pérez de Val, Bernat; López-Soria, Sergio; Nofrarías, Miquel; Martín, Maite; Vordermeier, H. Martin; Villarreal-Ramos, Bernardo; Romera, Nadine; Escobar, Manel; Solanes, David; Cardona, Pere-Joan; Domingo, Mariano

    2011-01-01

    Caprine tuberculosis (TB) has increased in recent years, highlighting the need to address the problem the infection poses in goats. Moreover, goats may represent a cheaper alternative for testing of prototype vaccines in large ruminants and humans. With this aim, a Mycobacterium caprae infection model has been developed in goats. Eleven 6-month-old goats were infected by the endobronchial route with 1.5 × 103 CFU, and two other goats were kept as noninfected controls. The animals were monitored for clinical and immunological parameters throughout the experiment. After 14 weeks, the goats were euthanized, and detailed postmortem analysis of lung lesions was performed by multidetector computed tomography (MDCT) and direct observation. The respiratory lymph nodes were also evaluated and cultured for bacteriological analysis. All infected animals were positive in a single intradermal comparative cervical tuberculin (SICCT) test at 12 weeks postinfection (p.i.). Gamma interferon (IFN-γ) antigen-specific responses were detected from 4 weeks p.i. until the end of the experiment. The humoral response to MPB83 was especially strong at 14 weeks p.i. (13 days after SICCT boost). All infected animals presented severe TB lesions in the lungs and associated lymph nodes. M. caprae was recovered from pulmonary lymph nodes in all inoculated goats. MDCT allowed a precise quantitative measure of TB lesions. Lesions in goats induced by M. caprae appeared to be more severe than those induced in cattle by M. bovis over a similar period of time. The present work proposes a reliable new experimental animal model for a better understanding of caprine tuberculosis and future development of vaccine trials in this and other species. PMID:21880849

  12. Peste des Petits Ruminants Virus Tissue Tropism and Pathogenesis in Sheep and Goats following Experimental Infection

    Science.gov (United States)

    Truong, Thang; Boshra, Hani; Embury-Hyatt, Carissa; Nfon, Charles; Gerdts, Volker; Tikoo, Suresh; Babiuk, Lorne A.; Kara, Pravesh; Chetty, Thireshni; Mather, Arshad; Wallace, David B.; Babiuk, Shawn

    2014-01-01

    Peste des petits ruminants (PPR) is a viral disease which primarily affects small ruminants, causing significant economic losses for the livestock industry in developing countries. It is endemic in Saharan and sub-Saharan Africa, the Middle East and the Indian sub-continent. The primary hosts for peste des petits ruminants virus (PPRV) are goats and sheep; however recent models studying the pathology, disease progression and viremia of PPRV have focused primarily on goat models. This study evaluates the tissue tropism and pathogenesis of PPR following experimental infection of sheep and goats using a quantitative time-course study. Upon infection with a virulent strain of PPRV, both sheep and goats developed clinical signs and lesions typical of PPR, although sheep displayed milder clinical disease compared to goats. Tissue tropism of PPRV was evaluated by real-time RT-PCR and immunohistochemistry. Lymph nodes, lymphoid tissue and digestive tract organs were the predominant sites of virus replication. The results presented in this study provide models for the comparative evaluation of PPRV pathogenesis and tissue tropism in both sheep and goats. These models are suitable for the establishment of experimental parameters necessary for the evaluation of vaccines, as well as further studies into PPRV-host interactions. PMID:24498032

  13. Evaluation of the therapeutic effect of potassium permanganate at early stages of an experimental acute infection of Flavobacterium columnare in channel catfish (Ictalurus punctatus)

    Science.gov (United States)

    The efficacy of potassium permanganate (KMnO4) against early stages of an experimental acute infection of Flavobacterium columnare in channel catfish (Ictalurus punctatus) was evaluated. Fish were experimentally challenged, by waterborne exposure for 2 h to F. columnare after cutaneous abrasion, an...

  14. Distribution patterns and predilection muscles of Trichinella zimbabwensis larvae in experimentally infected Nile crocodiles (Crocodylus niloticus Laurenti

    Directory of Open Access Journals (Sweden)

    Louis J. La Grange

    2014-02-01

    Full Text Available No controlled studies have been conducted to determine the predilection muscles of Trichinella zimbabwensis larvae in Nile crocodiles (Crocodylus niloticus or the influence of infection intensity on the distribution of the larvae in crocodiles. The distribution of larvae in muscles of naturally infected Nile crocodiles and experimentally infected caimans (Caiman crocodilus and varans (Varanus exanthematicus have been reported in literature. To determine the distribution patterns of T. zimbabwensis larvae and predilection muscles, 15 crocodiles were randomly divided into three cohorts of five animals each, representing high infection (642 larvae/kg of bodyweight average, medium infection (414 larvae/kg of bodyweight average and low infection (134 larvae/kg of bodyweight average cohorts. In the high infection cohort, high percentages of larvae were observed in the triceps muscles (26% and hind limb muscles (13%. In the medium infection cohort, high percentages of larvae were found in the triceps muscles (50%, sternomastoid (18% and hind limb muscles (13%. In the low infection cohort, larvae were mainly found in the intercostal muscles (36%, longissimus complex (27%, forelimb muscles (20% and hind limb muscles (10%. Predilection muscles in the high and medium infection cohorts were similar to those reported in naturally infected crocodiles despite changes in infection intensity. The high infection cohort had significantly higher numbers of larvae in the sternomastoid, triceps, intercostal, longissimus complex, external tibial flexor, longissimus caudalis and caudal femoral muscles (p < 0.05 compared with the medium infection cohort. In comparison with the low infection cohort, the high infection cohort harboured significantly higher numbers of larvae in all muscles (p < 0.05 except for the tongue. The high infection cohort harboured significantly higher numbers of larvae (p < 0.05 in the sternomastoid, triceps, intercostal, longissimus complex

  15. [Five steps to decreasing nosocomial infections in large immature premature infants: A quasi-experimental study].

    Science.gov (United States)

    García González, Ana; Leante Castellanos, José Luis; Fuentes Gutiérrez, Carmen; Lloreda García, José María; Fernández Fructuoso, José Ramón; Gómez Santos, Elisabet; García González, Verónica

    2017-07-01

    An evaluation is made of the impact of a series of five interventions on the incidence of hospital-related infections in a level iii neonatal unit. Quasi-experimental, pre-post intervention study, which included preterm infants weighing 1,500g at birth or delivered at <32 weeks gestation, admitted in the 12 months before and after the measures were implemented (January 2014). The measures consisted of: optimising hand washing, following a protocol for insertion and handling of central intravenous catheters, encouraging breastfeeding; applying a protocol for rational antibiotic use, and establishing a surveillance system for multi-resistant bacteria. The primary endpoint was to assess the incidence of hospital-acquired infections before and after implementing the interventions. Thirty-three matched patients were included in each period. There was an incidence of 8.7 and 2.7 hospital-related infections/1,000 hospital stay days in the pre- and post-intervention periods, respectively (P<.05). Additionally, patients in the treatment group showed a statistically-significant decrease in days on mechanical ventilation, use of blood products, and vasoactive drugs. The strategy, based on implementing five specific measures in a unit with a high rate of hospital-related infections, proved effective in reducing their incidence. This reduction could contribute to lowering the use of mechanical ventilation, blood products, and vasoactive drugs. Copyright © 2016 Asociación Española de Pediatría. Publicado por Elsevier España, S.L.U. All rights reserved.

  16. Assessment of Domestic Goats as Models for Experimental and Natural Infection with the North American Isolate of Rickettsia slovaca.

    Science.gov (United States)

    Lukovsky-Akhsanov, Nicole; Keating, M Kelly; Spivey, Pamela; Lathrop, George W; Powell, Nathaniel; Levin, Michael L

    2016-01-01

    Rickettsia slovaca is a tick-borne human pathogen that is associated with scalp eschars and neck lymphadenopathy known as tick-borne lymphadenopathy (TIBOLA) or Dermacentor-borne necrosis erythema and lymphadenopathy (DEBONEL). Originally, R. slovaca was described in Eastern Europe, but since recognition of its pathogenicity, human cases have been reported throughout Europe. European vertebrate reservoirs of R. slovaca remain unknown, but feral swine and domestic goats have been found infected or seropositive for this pathogen. Recently, a rickettsial pathogen identical to R. slovaca was identified in, and isolated from, the American dog tick, Dermacentor variabilis. In previous experimental studies, this organism was found infectious to guinea pigs and transovarially transmissible in ticks. In this study, domestic goats (Capra hircus) were experimentally inoculated with the North American isolate of this R. slovaca-like agent to assess their reservoir competence-the ability to acquire the pathogens and maintain transmission between infected and uninfected ticks. Goats were susceptible to infection as demonstrated by detection of the pathogen in skin biopsies and multiple internal tissues, but the only clinical sign of illness was transient fever noted in three out of four goats, and reactive lymphoid hyperplasia. On average, less than 5% of uninfected ticks acquired the pathogen while feeding upon infected goats. Although domestic goats are susceptible to the newly described North American isolate of R. slovaca, they are likely to play a minor role in the natural transmission cycle of this pathogen. Our results suggest that goats do not propagate the North American isolate of R. slovaca in peridomestic environments and clinical diagnosis of infection could be difficult due to the brevity and mildness of clinical signs. Further research is needed to elucidate the natural transmission cycle of R. slovaca both in Europe and North America, as well as to identify a

  17. Experimental infection of macaques with a wild water bird-derived highly pathogenic avian influenza virus (H5N1.

    Directory of Open Access Journals (Sweden)

    Tomoko Fujiyuki

    Full Text Available Highly pathogenic avian influenza virus (HPAIV continues to threaten human health. Non-human primate infection models of human influenza are desired. To establish an animal infection model with more natural transmission and to determine the pathogenicity of HPAIV isolated from a wild water bird in primates, we administered a Japanese isolate of HPAIV (A/whooper swan/Hokkaido/1/2008, H5N1 clade 2.3.2.1 to rhesus and cynomolgus monkeys, in droplet form, via the intratracheal route. Infection of the lower and upper respiratory tracts and viral shedding were observed in both macaques. Inoculation of rhesus monkeys with higher doses of the isolate resulted in stronger clinical symptoms of influenza. Our results demonstrate that HPAIV isolated from a water bird in Japan is pathogenic in monkeys by experimental inoculation, and provide a new method for HPAIV infection of non-human primate hosts, a good animal model for investigation of HPAIV pathogenicity.

  18. Immunodiagnosis of systemic aspergillosis. I. Antigenemia detected by radioimmunoassay in experimental infection. [/sup 125/I tracer technique

    Energy Technology Data Exchange (ETDEWEB)

    Weiner, M.H.; Coats-Stephen, M.

    1979-01-01

    Because systemic aspergillosis is difficult to diagnose ante mortem, a study to improve immunodiagnosis was undertaken in a rabbit model of disseminated infection. We found that the predominant humoral response of infected animals was directed against four Aspergillus antigens identified by crossed immunoelectrophoresis. One of these antigens, a cell-wall carbohydrate, was purified by gel-filtration chromatography and was used to develop a radiommunoassay. The sensitivity of this assay was increased by testing for serum-bound antigen as well as for free antigen. When the sensitivity of the RIA was evaluated in the animal model, antigenemia was detected in 78% of 51 rabbits with disseminated infection and ante mortem in 86% of 42 rabbits with lethal infection. By contrast, with immunoprecipitin analysis only eight of 51 rabbits were positive for antigen, and six of 51 rabbits were positive for Aspergillus antibody. The specificity of the RIA was also tested. Negative controls for antigen included sera from 76 normal rabbits and sera from 25 rabbits with systemic candidiasis. The Candida control group is pertinent because 48% of these rabbits had specific Candida antigenemia detected by a mannan RIA. This study demonstrates that Aspergillus antigenemia occurs during the course of experimental disseminated aspergillosis and illustrates the potential of an Aspergillus antigen RIA for sensitive, specific immunodiagnosis of human infections.

  19. A novel approach to parasite population genetics: experimental infection reveals geographic differentiation, recombination and host-mediated population structure in Pasteuria ramosa, a bacterial parasite of Daphnia.

    Science.gov (United States)

    Andras, J P; Ebert, D

    2013-02-01

    The population structure of parasites is central to the ecology and evolution of host-parasite systems. Here, we investigate the population genetics of Pasteuria ramosa, a bacterial parasite of Daphnia. We used natural P. ramosa spore banks from the sediments of two geographically well-separated ponds to experimentally infect a panel of Daphnia magna host clones whose resistance phenotypes were previously known. In this way, we were able to assess the population structure of P. ramosa based on geography, host resistance phenotype and host genotype. Overall, genetic diversity of P. ramosa was high, and nearly all infected D. magna hosted more than one parasite haplotype. On the basis of the observation of recombinant haplotypes and relatively low levels of linkage disequilibrium, we conclude that P. ramosa engages in substantial recombination. Isolates were strongly differentiated by pond, indicating that gene flow is spatially restricted. Pasteuria ramosa isolates within one pond were segregated completely based on the resistance phenotype of the host-a result that, to our knowledge, has not been previously reported for a nonhuman parasite. To assess the comparability of experimental infections with natural P. ramosa isolates, we examined the population structure of naturally infected D. magna native to one of the two source ponds. We found that experimental and natural infections of the same host resistance phenotype from the same source pond were indistinguishable, indicating that experimental infections provide a means to representatively sample the diversity of P. ramosa while reducing the sampling bias often associated with studies of parasite epidemics. These results expand our knowledge of this model parasite, provide important context for the large existing body of research on this system and will guide the design of future studies of this host-parasite system. © 2012 Blackwell Publishing Ltd.

  20. Desenvolvimento de um modelo experimental de falha óssea infectada na ulna de coelhos Development of an experimental model of infected bone void in the ulna of rabbits

    Directory of Open Access Journals (Sweden)

    Matheus Lemos Azi

    2012-01-01

    Full Text Available OBJETIVO: Desenvolver um modelo experimental que permita estudar a regeneração de grandes falhas ósseas em condições de infecção. MÉTODO: Falhas ósseas segmentares de 15mm foram criadas cirurgicamente na ulna de 12 coelhos e inoculadas com 5x10(8 unidades formadoras de colônia (UFC de S. aureus. O desbridamento da infecção foi realizado duas semanas após, seguida da aplicação sistêmica de gentamicina por quatro semanas. Os animais foram acompanhados por um período de 12 semanas para avaliação do controle da infecção e da regeneração óssea. RESULTADOS: A regeneração espontânea foi inferior a 25% do defeito na avaliação radiográfica e histológica. CONCLUSÃO: A Falha óssea infectada de 15mm na ulna de coelhos é incapaz de alcançar a regeneração completa sem tratamentos adicionais. Nível de Evidência V, Estudo experimental.OBJECTIVE: Develop a model that allowed the study of bone regeneration in infection conditions. METHOD: A 15 mm defect was surgically created in the rabbit ulna and inoculated with 5x10(8 colony-forming units (CFU of S. aureus. Surgical debridement was performed two weeks after and systemic gentamicin was administered for four weeks. Animals were followed up to 12 weeks to evaluate infection control and bone regeneration. RESULT: Bone regeneration was inferior to 25% of the defect in radiological and histological analysis. CONCLUSION: Infected bone defect of 15 mm in the rabbit ulna was unable to achieve full regeneration without further treatment. Level of Evidence V, Experimental Study.

  1. EXPERIMENTAL INFECTION BY Trypanosoma vivax IN GOATS INFECÇÃO EXPERIMENTAL EM CAPRINOS COM Trypanosoma vivax

    Directory of Open Access Journals (Sweden)

    Francisco David Nascimento Sousa

    2008-10-01

    Full Text Available

    Four goats were infected intravenously with 1.0 mL of cattle blood containing about 1.25 x 105 Trypanosoma vivax derived from spontaneous outbreak in cattle at Catolé do Rocha city, Paraíba, Brazil. Other four goats were used as controls. Parasitemia and body temperature were determined daily for 40 days. Animals were weighted each 7 days, and blood samples for blood cells counts were collected each 5 days. It was obtained a sample of liquor from each animal before death; cerebrospinal fluid samples were submitted to biochemical and cytological evaluations, density determination and parasite detection. A positive correlation was found between body temperature and parasitemia in infected animals. These animals presented anemia, leukopenia, hypoglycemia, decreased serum levels of total proteins and cholesterol, and nervous symptoms. Examination of cerebrospinal fluid resulted in decrease of glucose levels and increase in lactate dehydrogenase, cell counts and presence of the parasite. At necropsy it was found pale carcass, generalized infartation of lymphonodes, pulmonary edema, and liquid accumulation of pericardium. Histological changes were characterized by interstitial pneumonia, miocarditis, cardiac fibrosis, meningitis, and encephalitis. All observed changes confirm patogenicity of T. vivax.

    KEY WORDS: Experimental infection, trypanosomiasis, patogenicity.

    Quatro caprinos foram infectados experimentalmente por via intravenosa com 1,0 ml de sangue contendo aproximadamente 1,25 x 105 tripanossomas/ml, utilizando-se um isolado de Trypanosoma vivax de bovinos infectados naturalmente no município de Catolé do Rocha, Paraíba. A parasitemia e a temperatura foram determinadas diariamente durante quarenta dias. A cada cinco dias realizaram-se coletas de sangue para hemograma e análise bioquímica sérica. Antes do

  2. Highly (H5N1 and low (H7N2 pathogenic avian influenza virus infection in falcons via nasochoanal route and ingestion of experimentally infected prey.

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    Kateri Bertran

    Full Text Available An experimental infection with highly pathogenic avian influenza (HPAI and low pathogenic avian influenza (LPAI viruses was carried out on falcons in order to examine the effects of these viruses in terms of pathogenesis, viral distribution in tissues and viral shedding. The distribution pattern of influenza virus receptors was also assessed. Captive-reared gyr-saker (Falco rusticolus x Falco cherrug hybrid falcons were challenged with a HPAI H5N1 virus (A/Great crested grebe/Basque Country/06.03249/2006 or a LPAI H7N2 virus (A/Anas plathyrhynchos/Spain/1877/2009, both via the nasochoanal route and by ingestion of previously infected specific pathogen free chicks. Infected falcons exhibited similar infection dynamics despite the different routes of exposure, demonstrating the effectiveness of in vivo feeding route. H5N1 infected falcons died, or were euthanized, between 5-7 days post-infection (dpi after showing acute severe neurological signs. Presence of viral antigen in several tissues was confirmed by immunohistochemistry and real time RT-PCR (RRT-PCR, which were generally associated with significant microscopical lesions, mostly in the brain. Neither clinical signs, nor histopathological findings were observed in any of the H7N2 LPAI infected falcons, although all of them had seroconverted by 11 dpi. Avian receptors were strongly present in the upper respiratory tract of the falcons, in accordance with the consistent oral viral shedding detected by RRT-PCR in both H5N1 HPAI and H7N2 LPAI infected falcons. The present study demonstrates that gyr-saker hybrid falcons are highly susceptible to H5N1 HPAI virus infection, as previously observed, and that they may play a major role in the spreading of both HPAI and LPAI viruses. For the first time in raptors, natural infection by feeding on infected prey was successfully reproduced. The use of avian prey species in falconry husbandry and wildlife rehabilitation facilities could put valuable birds

  3. Highly (H5N1) and low (H7N2) pathogenic avian influenza