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Richard W. Gerhold - One of the best experts on this subject based on the ideXlab platform.
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Detection of Trichomonas gallinae in Wild Birds Admitted to a Rehabilitation Center, Florida, USA
Journal of wildlife diseases, 2020Co-Authors: Maria C. Spriggs, Kathryn E. Purple, Richard W. GerholdAbstract:We screened raptors (n=188) and columbids (n=2) presenting to a Tampa, Florida, US rehabilitation center from June 2016 to April 2017 for Trichomonas gallinae. One juvenile Cooper's Hawk (Accipiter cooperii) was culture- and PCR-positive and sequencing identified as genotype E1. A subset of culture-negative (10%, n=19) samples were negative via PCR.
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Improved subtyping affords better discrimination of Trichomonas gallinae strains and suggests hybrid lineages.
Infection genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases, 2019Co-Authors: Abdulwahed F. Alrefaei, Richard W. Gerhold, Diana Bell, Johanna L. Nader, Kevin M. TylerAbstract:Trichomonas gallinae is a protozoan pathogen that causes avian trichomonosis typically associated with columbids (canker) and birds of prey (frounce) that predate on them, and has recently emerged as an important cause of passerine disease. An archived panel of DNA from North American (USA) birds used initially to establish the ITS ribotypes was reanalysed using Iron hydrogenase (FeHyd) gene sequences to provide an alphanumeric subtyping scheme with improved resolution for strain discrimination. Thirteen novel subtypes of T. gallinae using FeHyd gene as the subtyping locus are described. Although the phylogenetic topologies derived from each single marker are complementary, they are not entirely congruent. This may reflect the complex genetic histories of the isolates analysed which appear to contain two major lineages and several that are hybrid. This new analysis consolidates much of the phylogenetic signal generated from the ITS ribotype and provides additional resolution for discrimination of T. gallinae strains. The single copy FeHyd gene provides higher resolution genotyping than ITS ribotype alone. It should be used where possible as an additional, single-marker subtyping tool for cultured isolates.
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Artificially decreased dissolved oxygen increases the persistence of Trichomonas gallinae in water.
International journal for parasitology. Parasites and wildlife, 2019Co-Authors: Kathryn E. Purple, Todd Michael Amacker, Chauntelle Williams, Richard W. GerholdAbstract:Water containing organic material has been shown to increase the persistence of the avian pathogenic protozoa, Trichomonas gallinae. We hypothesized that the decrease in dissolved oxygen due to microbes in the organic material could increase persistence of the microaerophilic trichomonads. Using simulated birdbaths, we determined 1) the levels of dissolved oxygen in distilled water with various amounts of organic material, 2) the concentration of the oxygen-scavenging enzyme Oxyrase® needed to achieve the dissolved oxygen levels obtained in organic material contaminated water, and finally, 3) the persistence of two T. gallinae isolates in Oxyrase®-supplemented water. An average of 9.6% dissolved oxygen was obtained with the addition of 15 g organic material to 500 ml of distilled water, whereas organic material-free water had 86.2% dissolved oxygen. The addition of 0.5% and 1.0% (vol/vol) Oxyrase® to organic material-free water yielded dissolved oxygen of 18.6% and 6.9%, respectively. Using 0.5% and 1.0% concentrations of Oxyrase®, we evaluated the persistence of two trichomonad isolates by inoculating ∼1 million trichomonads into 500 ml distilled water in triplicate. At various time-points, 0.5 ml aliquots of trichomonad-inoculated water were obtained and placed into Hollander Fluid media, incubated at 37 °C, and read by light microscopy every other day for 5 days. In our 1% Oxyrase® treatments, the longest recorded persistence of broad-winged hawk 1 increased from the previously reported 4hrs to 30hrs and Cooper's hawk 4 from 16hrs to 30hrs. These results indicate that the mechanism for organic material-mediated trichomonad persistence is associated with decreased dissolved oxygen, further demonstrating the importance of keeping birdbaths free of organic debris to discourage trichomonad persistence.
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prevalence and risk factors of Trichomonas gallinae and trichomonosis in golden eagle aquila chrysaetos nestlings in western north america
Journal of Wildlife Diseases, 2018Co-Authors: Benjamin M Dudek, Kathryn E. Purple, Richard W. Gerhold, Michael N Kochert, Joseph G Barnes, Peter H Bloom, Joseph M Papp, Kenneth V Jacobson, Charles R Preston, Chris R VennumAbstract:Abstract Avian trichomonosis, caused by the protozoan Trichomonas gallinae, affects bird-eating raptors worldwide. Raptors can develop trichomonosis by feeding on infected prey, particularly Rock Pigeons (Columba livia), which are a reservoir for T. gallinae. Raptors may be particularly vulnerable to T. gallinae infection in degraded habitats, where changes in resources may cause raptors to switch from foraging on native prey to synanthropic avian species such as Rock Pigeons. Golden Eagles (Aquila chrysaetos) typically forage on mammals; however, habitat across much of their range is experiencing degradation through changes in land use, climate, and human encroachment. In 2015, we examined the prevalence of T. gallinae infection in Golden Eagle nestlings across western North America and conducted an intensive study on factors associated with T. gallinae infection and trichomonosis in southwestern Idaho. We found T. gallinae infection in 13% (12/96) of eagle nestlings across 10 western states and in 41% (...
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Trichomonas gallinae persistence in water with ph and organic material variance
Comparative Parasitology, 2016Co-Authors: Forrest Sturgill, Richard W. GerholdAbstract:ABSTRACT: Trichomonas gallinae is a protozoan parasite commonly found in domestic turkeys, columbids, and passerines. Trichomonosis can cause significant morbidity and mortality in these species and has been associated with contaminated bird feeders and waterers. Previously, we found that T. gallinae persisted up to 1 hr in distilled water with organic material; however, it was recognized that pH may play a role in the persistence of isolates of T. gallinae. To examine the effects of pH, we inoculated 1 × 106 trichomonads into plastic containers in 4 different treatments: 500 ml clean distilled water at pH 5.0 or 7.0, or 500 ml distilled water at pH 5.0 or 7.0 plus 15 g organic matter. Aliquots of 0.5 ml were collected from each container at 0, 0.5, 1, 5, 10, 20, 60 min, or 24 hr and inoculated into a flask with Diamond’s media, incubated at 37°C, and examined daily for 6 d. Maximum persistence of T. gallinae for pH 7 was 60 min, whereas the maximum persistence at pH 5.0 was 20 min. Survival was best in ...
Tiana Tasca - One of the best experts on this subject based on the ideXlab platform.
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Cytotoxicity of solubilization vehicles for Trichomonas gallinae and TriTrichomonas foetus measured by the resazurin microtiter assay
Veterinary Parasitology, 2009Co-Authors: Mariana Duarte, G. A. De Carli, Raquel Brandt Giordani, José Angelo S. Zuanazzi, Alexandre José Macedo, Tiana TascaAbstract:The parasites TriTrichomonas foetus and Trichomonas gallinae present veterinary and economic importance since they cause bovine and avian trichomonosis, respectively. The absence of a specific treatment and the necessity of effective and safe drugs against these etiologic agents have stimulated the search for new antiprotozoal drugs with high activity, low toxicity to the animal, and low cost. Screening of potential antiprotozoal molecules is currently a common practice and different kinds of organic solvents and surfactant vehicles are used, since many bioactive compounds have low water solubility. Thus, it is important to determine the toxicity and to provide the minimal inhibitory concentration (MIC) values of the most common solubilization vehicles used in biological activity in vitro evaluation: ethanol, methanol, isopropanol, acetone, DMSO, tween 20 and tween 80. The assays were conducted employing the resazurin microtiter assay, which demonstrated a rapid, safe, and quantitative method for the in vitro determination of T. foetus and T. gallinae trophozoites viability. In summary, all solvents and surfactants, except ethanol, could be used in cytotoxicity assays against T. foetus, and acetone, tween 20 and tween 80 are the preferential vehicles for in vitro analysis of potential bioactive compounds against T. gallinae, though these must be used with caution.
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Analysis of microtubule cytoskeleton distribution using a fluorescent taxoid in two trichomonadid protozoa: Trichomonas gallinae and TriTrichomonas foetus.
Experimental parasitology, 2007Co-Authors: Patrícia De Brum Vieira, Tiana Tasca, Fernanda Pires Borges, Bárbara Gottardi, Cristiane Dos Santos Stuepp, Anne Brandolt Larré, G. A. De CarliAbstract:Abstract Trichomonas gallinae and TriTrichomonas foetus are flagellated parasitic protozoa of the upper digestive tract of birds and the urogenital tract of cattle, respectively. Both of these species are important in the veterinary field, due to the fact that they cause significant economic losses. Therefore, we investigated the morphology of these parasites by studying microtubule cytoskeleton organization. FLUTAX-2, an active fluorescent derivative of Taxol, was used in this study. This fluorescent taxoid binds to polymerized αβ-tubulin dimers. Our results showed that FLUTAX-2 was able to bind to and stabilize microtubules of intact T. gallinae and T. foetus trophozoites, allowing the microtubular cytoskeleton to be easily observed by fluorescence microscopy. T. foetus and T. gallinae had no differences in their FLUTAX-2 binding profiles. Further studies may allow this technique to be improved, and it may possibly be used as a routine laboratory method for the diagnosis of avian and bovine trichomonosis.
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Characterization of an ecto-5'-nucleotidase (EC 3.1.3.5) activity in intact trophozoites of Trichomonas gallinae.
Veterinary parasitology, 2006Co-Authors: Fernanda Pires Borges, G. A. De Carli, Tiana Tasca, Bárbara Gottardi, Cristiane Dos Santos Stuepp, Anne Brandolt Larré, Carla Denise BonanAbstract:This study describes the enzymatic properties of an ecto-5'-nucleotidase in Trichomonas gallinae. The enzyme hydrolyzes nucleoside monophosphates at pH 7.2 and is activated by divalent cations, such as magnesium. Ecto-5'-nucleotidase activity was insensitive to levamisole, tetramisole (alkaline phosphatase inhibitors), and AMPCP (adenosine 5'-[alpha,beta-methylene]diphosphate), an ecto-5'-nucleotidase inhibitor, whereas 0.1mM ammonium molybdate (considered a potent inhibitor of 5'-nucleotidase activity) completely inhibited the enzyme activity. The apparent K(M) (Michaelis constant) and Vmax (maximum velocity) values for Mg2+-AMP were 466+/-57 microM and 3.7+/-0.59 nmolPi/min/10(6) trichomonads, respectively. Considering that trichomonads lack the ability to synthesize purines and pyrimidines de novo, the presence of an ecto-5'-nucleotidase in intact trophozoites of T. gallinae could be important in regulating the extracellular nucleotide levels and generating adenosine, essential for the survival strategies of the parasite.
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Scanning electron microscopy study of Trichomonas gallinae.
Veterinary parasitology, 2003Co-Authors: Tiana Tasca, Geraldo A De CarliAbstract:A scanning electron microscopy (SEM) study of Trichomonas gallinae (Rivolta, 1878), provided more information about the morphology of this flagellated protozoan. SEM showed the morphological features of the trophozoites; the emergence of the anterior flagella, the structure of the undulating membrane, the position and shape of the pelta, axostyle and posterior flagellum. Of special interest were the pseudocyst forms.
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Trichomonas gallinae: a possible contact-dependent mechanism in the hemolytic activity.
Veterinary parasitology, 2002Co-Authors: G. A. De Carli, Tiana TascaAbstract:Abstract The in vitro hemolytic activity of Trichomonas gallinae was investigated. The parasite was tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of six adult animals of different species (rabbit, rat, chicken, horse, bovine, and sheep). Results showed that T. gallinae lysed all human erythrocytes groups, as well as rabbit, rat, chicken, horse, bovine and sheep erythrocytes. No hemolysin released by the parasites could be identified. Hemolysis did not occur with trichomonad culture supernatants, with sonicated extracts of T. gallinae, or with killed organisms. The scanning electron microscopy (SEM) showed that the erythocytes adhered to the parasite surface and were phagocytosed. These observations suggest that the contact between T. gallinae and erythrocytes may be an important mechanism in the injury caused to the erythrocytes. The hemolytic activity of T. gallinae may be an efficient means of obtaining nutrients for the parasite and allow the investigation of the mechanism used by T. gallinae to damage cellular membranes.
Kathryn E. Purple - One of the best experts on this subject based on the ideXlab platform.
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Detection of Trichomonas gallinae in Wild Birds Admitted to a Rehabilitation Center, Florida, USA
Journal of wildlife diseases, 2020Co-Authors: Maria C. Spriggs, Kathryn E. Purple, Richard W. GerholdAbstract:We screened raptors (n=188) and columbids (n=2) presenting to a Tampa, Florida, US rehabilitation center from June 2016 to April 2017 for Trichomonas gallinae. One juvenile Cooper's Hawk (Accipiter cooperii) was culture- and PCR-positive and sequencing identified as genotype E1. A subset of culture-negative (10%, n=19) samples were negative via PCR.
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Artificially decreased dissolved oxygen increases the persistence of Trichomonas gallinae in water.
International journal for parasitology. Parasites and wildlife, 2019Co-Authors: Kathryn E. Purple, Todd Michael Amacker, Chauntelle Williams, Richard W. GerholdAbstract:Water containing organic material has been shown to increase the persistence of the avian pathogenic protozoa, Trichomonas gallinae. We hypothesized that the decrease in dissolved oxygen due to microbes in the organic material could increase persistence of the microaerophilic trichomonads. Using simulated birdbaths, we determined 1) the levels of dissolved oxygen in distilled water with various amounts of organic material, 2) the concentration of the oxygen-scavenging enzyme Oxyrase® needed to achieve the dissolved oxygen levels obtained in organic material contaminated water, and finally, 3) the persistence of two T. gallinae isolates in Oxyrase®-supplemented water. An average of 9.6% dissolved oxygen was obtained with the addition of 15 g organic material to 500 ml of distilled water, whereas organic material-free water had 86.2% dissolved oxygen. The addition of 0.5% and 1.0% (vol/vol) Oxyrase® to organic material-free water yielded dissolved oxygen of 18.6% and 6.9%, respectively. Using 0.5% and 1.0% concentrations of Oxyrase®, we evaluated the persistence of two trichomonad isolates by inoculating ∼1 million trichomonads into 500 ml distilled water in triplicate. At various time-points, 0.5 ml aliquots of trichomonad-inoculated water were obtained and placed into Hollander Fluid media, incubated at 37 °C, and read by light microscopy every other day for 5 days. In our 1% Oxyrase® treatments, the longest recorded persistence of broad-winged hawk 1 increased from the previously reported 4hrs to 30hrs and Cooper's hawk 4 from 16hrs to 30hrs. These results indicate that the mechanism for organic material-mediated trichomonad persistence is associated with decreased dissolved oxygen, further demonstrating the importance of keeping birdbaths free of organic debris to discourage trichomonad persistence.
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prevalence and risk factors of Trichomonas gallinae and trichomonosis in golden eagle aquila chrysaetos nestlings in western north america
Journal of Wildlife Diseases, 2018Co-Authors: Benjamin M Dudek, Kathryn E. Purple, Richard W. Gerhold, Michael N Kochert, Joseph G Barnes, Peter H Bloom, Joseph M Papp, Kenneth V Jacobson, Charles R Preston, Chris R VennumAbstract:Abstract Avian trichomonosis, caused by the protozoan Trichomonas gallinae, affects bird-eating raptors worldwide. Raptors can develop trichomonosis by feeding on infected prey, particularly Rock Pigeons (Columba livia), which are a reservoir for T. gallinae. Raptors may be particularly vulnerable to T. gallinae infection in degraded habitats, where changes in resources may cause raptors to switch from foraging on native prey to synanthropic avian species such as Rock Pigeons. Golden Eagles (Aquila chrysaetos) typically forage on mammals; however, habitat across much of their range is experiencing degradation through changes in land use, climate, and human encroachment. In 2015, we examined the prevalence of T. gallinae infection in Golden Eagle nestlings across western North America and conducted an intensive study on factors associated with T. gallinae infection and trichomonosis in southwestern Idaho. We found T. gallinae infection in 13% (12/96) of eagle nestlings across 10 western states and in 41% (...
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Survey for Trichomonas gallinae in Northern Bobwhites (Colinus virginianus) from the Rolling Plains Ecoregion, Oklahoma and Texas, USA
Journal of wildlife diseases, 2015Co-Authors: Andrea Bruno, Kathryn E. Purple, Richard W. Gerhold, Alan M. Fedynich, Dale RollinsAbstract:Northern Bobwhites (Colinus virginianus) have been in decline throughout the southeastern US. Prevalence of Trichomonas gallinae in wild bobwhites is unknown, although T. gallinae caused morbidity and mortality in experimentally infected bobwhites. Many species of Columbidae (pigeons and doves) in Texas are hosts to T. gallinae. Bobwhites potentially may become exposed to this protozoan through supplemental feed or water sources contaminated by columbids infected with T. gallinae. All of 506 bobwhites collected in Oklahoma and Texas, 2011-13, were PCR negative for T. gallinae. These data suggest T. gallinae is not contributing to the population decline of bobwhites in this region.
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Trichomonas gallinae Persistence in Four Water Treatments
Journal of wildlife diseases, 2015Co-Authors: Kathryn E. Purple, Jacob M. Humm, R. Brian Kirby, Christina G. Saidak, Richard W. GerholdAbstract:Trichomonas gallinae is a protozo- an parasite commonly found in columbids, passerines, and several raptor species. Although T. gallinae is thought to spread between individuals and across species through shared water sources, little research has been con- ducted regarding the persistence of T. gallinae in the environment. To determine the persis- tence of T. gallinae in various communal water sources, we inoculated 1310 6 trichomonads into 500 mL samples of distilled water, quarry water, bird bath water, and rain barrel water in two replicates. Aliquots of 0.5 mL were collected from each source at 21, 0, 15, 30, and 60 min; aliquots were incubated at 37 C and examined for trichomonads by light mi- croscopy for five consecutive days. Live tricho- monads were observed in all samples and at all sampling times except prior to inoculation (21 min). The pH of water sources ranged from an average of 5.9 to 7.4 postsampling. Our findings indicate that T. gallinae can persist for up to 60 min in various water treatments and thus be infectious for birds drinking T. gallinae-con- taminated water.
G. A. De Carli - One of the best experts on this subject based on the ideXlab platform.
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Cytotoxicity of solubilization vehicles for Trichomonas gallinae and TriTrichomonas foetus measured by the resazurin microtiter assay
Veterinary Parasitology, 2009Co-Authors: Mariana Duarte, G. A. De Carli, Raquel Brandt Giordani, José Angelo S. Zuanazzi, Alexandre José Macedo, Tiana TascaAbstract:The parasites TriTrichomonas foetus and Trichomonas gallinae present veterinary and economic importance since they cause bovine and avian trichomonosis, respectively. The absence of a specific treatment and the necessity of effective and safe drugs against these etiologic agents have stimulated the search for new antiprotozoal drugs with high activity, low toxicity to the animal, and low cost. Screening of potential antiprotozoal molecules is currently a common practice and different kinds of organic solvents and surfactant vehicles are used, since many bioactive compounds have low water solubility. Thus, it is important to determine the toxicity and to provide the minimal inhibitory concentration (MIC) values of the most common solubilization vehicles used in biological activity in vitro evaluation: ethanol, methanol, isopropanol, acetone, DMSO, tween 20 and tween 80. The assays were conducted employing the resazurin microtiter assay, which demonstrated a rapid, safe, and quantitative method for the in vitro determination of T. foetus and T. gallinae trophozoites viability. In summary, all solvents and surfactants, except ethanol, could be used in cytotoxicity assays against T. foetus, and acetone, tween 20 and tween 80 are the preferential vehicles for in vitro analysis of potential bioactive compounds against T. gallinae, though these must be used with caution.
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Characterization of an ecto-5'-nucleotidase (EC 3.1.3.5) activity in intact trophozoites of Trichomonas gallinae.
Veterinary parasitology, 2006Co-Authors: Fernanda Pires Borges, G. A. De Carli, Tiana Tasca, Bárbara Gottardi, Cristiane Dos Santos Stuepp, Anne Brandolt Larré, Carla Denise BonanAbstract:This study describes the enzymatic properties of an ecto-5'-nucleotidase in Trichomonas gallinae. The enzyme hydrolyzes nucleoside monophosphates at pH 7.2 and is activated by divalent cations, such as magnesium. Ecto-5'-nucleotidase activity was insensitive to levamisole, tetramisole (alkaline phosphatase inhibitors), and AMPCP (adenosine 5'-[alpha,beta-methylene]diphosphate), an ecto-5'-nucleotidase inhibitor, whereas 0.1mM ammonium molybdate (considered a potent inhibitor of 5'-nucleotidase activity) completely inhibited the enzyme activity. The apparent K(M) (Michaelis constant) and Vmax (maximum velocity) values for Mg2+-AMP were 466+/-57 microM and 3.7+/-0.59 nmolPi/min/10(6) trichomonads, respectively. Considering that trichomonads lack the ability to synthesize purines and pyrimidines de novo, the presence of an ecto-5'-nucleotidase in intact trophozoites of T. gallinae could be important in regulating the extracellular nucleotide levels and generating adenosine, essential for the survival strategies of the parasite.
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Trichomonas gallinae: a possible contact-dependent mechanism in the hemolytic activity.
Veterinary parasitology, 2002Co-Authors: G. A. De Carli, Tiana TascaAbstract:Abstract The in vitro hemolytic activity of Trichomonas gallinae was investigated. The parasite was tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of six adult animals of different species (rabbit, rat, chicken, horse, bovine, and sheep). Results showed that T. gallinae lysed all human erythrocytes groups, as well as rabbit, rat, chicken, horse, bovine and sheep erythrocytes. No hemolysin released by the parasites could be identified. Hemolysis did not occur with trichomonad culture supernatants, with sonicated extracts of T. gallinae, or with killed organisms. The scanning electron microscopy (SEM) showed that the erythocytes adhered to the parasite surface and were phagocytosed. These observations suggest that the contact between T. gallinae and erythrocytes may be an important mechanism in the injury caused to the erythrocytes. The hemolytic activity of T. gallinae may be an efficient means of obtaining nutrients for the parasite and allow the investigation of the mechanism used by T. gallinae to damage cellular membranes.
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Hemolytic activity of fresh isolates and clones of Trichomonas gallinae
Parasitología al día, 1999Co-Authors: Tiana Tasca, G. A. De CarliAbstract:In in vitro assay the hemolytic activity of live isolates and clones of Trichomonas gallinae was investigated. The fresh isolates and clones were tested against human erythrocytes of groups A, B, AB, and O, and against erythrocytes of six adult animals of different species (rabbit, rat, chicken, horse, bovine, and sheep). Results showed that each of the isolates and clones tested lysed all human blood groups, as well as rabbit, rat, chicken, horse, bovine and sheep erythrocytes. No hemolysin released by the parasites could be identified. Parasite culture supernatants of hemolysis assays, tested in the presence of human erythrocytes did not induce any hemolytic activity. Trichomonads culture supernatants from 18, 24, and 48 hr extracts of sonicated parasites or cold killed organisms tested with human erythrocytes did not exhibit hemolysis. Although an hemolytic activity of T. gallinae was clearly demonstrated, a relationship between hemolysis and cell pathogenicity is not yet clearly established.
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PREVALENCE OF Trichomonas gallinae FROM THE UPPER DIGESTIVE TRACT OF THE COMMON PIGEON, Columba livia IN THE SOUTHERN BRAZILIAN STATE, RIO GRANDE DO SUL
Parasitología al día, 1999Co-Authors: Tiana Tasca, G. A. De CarliAbstract:ABSTRACT Trichomonas gallinae (Rivolta, 1878) is a parasitic protozoan of the upper digestive tract and various organs of different avian groups, being common among the Columbids, was isolated described and its prevalence studies in the Southern Brazilian State, Rio Grande do Sul. The morphology study of the live specimens done by examination of fresh and stained specimens, showed that the T. gallinae isolated has the same morphological characteristics as the upper digestive tract trichomonads previously described by other authors
Michael Hess - One of the best experts on this subject based on the ideXlab platform.
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Multi-locus sequence typing confirms the clonality of Trichomonas gallinae isolates circulating in European finches.
Parasitology, 2013Co-Authors: Petra Ganas, Becki Lawson, Barbara Jaskulska, Marko Zadravec, Michael Hess, Ivana BilicAbstract:In recent years, Trichomonas gallinae emerged as the causative agent of an infectious disease of passerine birds in Europe leading to epidemic mortality of especially greenfinches Chloris chloris and chaffinches Fringilla coelebs . After the appearance of finch trichomonosis in the UK and Fennoscandia, the disease spread to Central Europe. Finch trichomonosis first reached Austria and Slovenia in 2012. In the present study the genetic heterogeneity of T. gallinae isolates from incidents in Austria and Slovenia were investigated and compared with British isolates. For this purpose comparative sequence analyses of the four genomic loci ITS1-5.8S-ITS2, 18S rRNA, rpb1 and Fe-hydrogenase were performed. The results corroborate that one clonal T. gallinae strain caused the emerging infectious disease within passerine birds and that the disease is continuing to spread in Europe. The same clonal strain was also found in a columbid bird from Austria. Additionally, the present study demonstrates clearly the importance of multi-locus sequence typing for discrimination of circulating T. gallinae strains.
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Axenization and optimization of in vitro growth of clonal cultures of TetraTrichomonas gallinarum and Trichomonas gallinae.
Experimental parasitology, 2009Co-Authors: Aziza Amin, C. Neubauer, Dieter Liebhart, Elvira Grabensteiner, Michael HessAbstract:A rapid and simple procedure was established to obtain clonal axenic cultures of TetraTrichomonas gallinarum and Trichomonas gallinae and to optimize their in vitro growth conditions. Medium 199 was used for axenization of two genetically different clones of T. gallinarum and T. gallinae. Six different media were used to optimize the growth behaviour of axenically grown parasites: Medium 199, TYM, TYI-S-33, Hollander fluid (HF), Trichomonas vaginalis (TV) and modified TV media. The highest cell yields for both axenic clones of T. gallinarum were obtained in modified TV medium without antibiotics. The maximum numbers of trophozoites of T. gallinae were obtained in an optimized HF medium. This study demonstrated that axenic cultures for T. gallinarum and T. gallinae could be obtained avoiding the migration technique through a V-tube. Following axenization and optimization, both clones of T. gallinarum and T. gallinae could be propagated both aerobically and anaerobically.
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Fine structure of the bird parasites Trichomonas gallinae and TetraTrichomonas gallinarum from cultures
Parasitology Research, 2009Co-Authors: Heinz Mehlhorn, Saleh Al-quraishy, Amin Aziza, Michael HessAbstract:The trophozoites of Trichomonas gallinae and TetraTrichomonas gallinarum were studied by means of light and electron microscopy after cloning and cultivating them axenically. T . gallinae trophozoites varied in shape reaching from ovoidal to pyriform and had a size of about 7–11 µm. They were provided with four free flagella and a fifth recurrent one, which did not become free at the posterior pole. The nucleus was ovoid, had a size of about 2.5–3 µm, and was situated closely below the basal bodies of the flagella. The axostyle consisted of a row of microtubules running from the region of the apical basal bodies to the posterior end of the cell. In addition to flagellated stages, which contained food vacuoles, hydrogenosomes, a costa-like structure, and glycogen granules besides lacunes of endoplasmic reticulum, spherical, nonflagellated, and cyst-like stages occurred. The trophozoites of T . gallinarum appeared mostly pear-shaped and ranged in size from 6 to 15 µm. They had also four free anterior flagella and a fifth recurrent one, which became free at the posterior pole in contrast to that of T . gallinae . Another clearly visible difference to T . gallinae was the occurrence of a sphere of lacunes of the endoplasmic reticulum surrounding in a regular distance the nucleus with its typical perinuclear membranes. Furthermore, the food vacuoles appeared very large. However, both species clearly differed from the trophozoites of Histomonas meleagridis .
