Fatal Toxoplasma gondii infection in the giant panda.

Science.gov (United States)

Ma, Hongyu; Wang, Zedong; Wang, Chengdong; Li, Caiwu; Wei, Feng; Liu, Quan

2015-01-01

Toxoplasma gondii can infect nearly all warm-blooded animals. We report an acute fatal T. gondii infection in the endangered giant panda (Ailuropoda melanoleuca) in a zoo in China, characterized by acute gastroenteritis and respiratory symptoms. T. gondii infection was confirmed by immunological and molecular methods. Multilocus nested PCR-RFLP revealed clonal type I at the SAG1 and c29-2 loci, clonal type II at the SAG2, BTUB, GRA6, c22-8, and L358 loci, and clonal type III at the alternative SAG2 and SAG3 loci, thus, a potential new genotype of T. gondii in the giant panda. Other possible pathogens were not detected. To our knowledge, this is the first report of clinical toxoplasmosis in a giant panda. © H. Ma et al., published by EDP Sciences, 2015.

Molecular Cloning Designer Simulator (MCDS: All-in-one molecular cloning and genetic engineering design, simulation and management software for complex synthetic biology and metabolic engineering projects

Directory of Open Access Journals (Sweden)

Zhenyu Shi

2016-12-01

Full Text Available Molecular Cloning Designer Simulator (MCDS is a powerful new all-in-one cloning and genetic engineering design, simulation and management software platform developed for complex synthetic biology and metabolic engineering projects. In addition to standard functions, it has a number of features that are either unique, or are not found in combination in any one software package: (1 it has a novel interactive flow-chart user interface for complex multi-step processes, allowing an integrated overview of the whole project; (2 it can perform a user-defined workflow of cloning steps in a single execution of the software; (3 it can handle multiple types of genetic recombineering, a technique that is rapidly replacing classical cloning for many applications; (4 it includes experimental information to conveniently guide wet lab work; and (5 it can store results and comments to allow the tracking and management of the whole project in one platform. MCDS is freely available from https://mcds.codeplex.com. Keywords: BioCAD, Genetic engineering software, Molecular cloning software, Synthetic biology, Workflow simulation and management

Using "Pseudomonas Putida xylE" Gene to Teach Molecular Cloning Techniques for Undergraduates

Science.gov (United States)

Dong, Xu; Xin, Yi; Ye, Li; Ma, Yufang

2009-01-01

We have developed and implemented a serial experiment in molecular cloning laboratory course for undergraduate students majored in biotechnology. "Pseudomonas putida xylE" gene, encoding catechol 2, 3-dioxygenase, was manipulated to learn molecular biology techniques. The integration of cloning, expression, and enzyme assay gave students…

SAG2 locus genotyping of Toxoplasma gondii in meat products of ...

African Journals Online (AJOL)

Toxoplasmosis is an infection caused by Toxoplasma gondii, an intracellular obligate parasite. Its transmission is usually attributed to ingestion of undercooked or raw meat. The aim of this study was the detection and genotyping of T. gondii in meat products using the molecular method in East Azerbaijan. DNA was ...

Serological and molecular detection of Toxoplasma gondii in Columba livia hunting pigeons of AL-Qadisiyah province

Directory of Open Access Journals (Sweden)

Hiba Riyadth Jameel Al-abodi

2017-07-01

Full Text Available Toxoplasma gondii were detected and diagnosed in 80 Columba livia birds at the province of Al-Qadisiyah during the period started from January 2016 until June 2016 using latex test and rapid cassette test ,Moreover molecular diagnosis with Polymerase Chain Rraction technique for identification of B1 gene had been also used Results showed that antibodies were detected in 11 samples out of 80 (13.75%. of Columba livia pigeons in Al-Qadisiyah province, Since significant highest (P≤0.05 titration were indicated at 1/80 (36.37%,whereas the lowest titer indicated at 1/40 (9.09 and According to type of antibodies in Columba livia toxoplasmosis suspected samples using rapid test cassette, results were indicated that Six samples out of 80 (7.5% were found positive. However IgG, IgM and IgG plus IgM were found in (33.34%,(16.66 and (50 respectively with no significance difference, furthermore results of PCR technique for detection of B1 gene revealed that 5% of samples from the Columba livia were only found positive ,Moreover the B1 gene were had a molecular weight of the private 399bp.It had been concluded that Columba livia birds were found infected with T.gondii with possible transmission to human being .

Molecular cloning and characterization of pathogenesis-related ...

African Journals Online (AJOL)

We described the cloning and characterization of pathogenesis-related protein 5 gene in maize, named ZmPR5 (GenBank Accession Number: HM230665). Molecular and bioinformatic analyses of ZmPR5 revealed an open reading frame (ORF) of 525 bp, encoding a protein of 175 amino acids (aa) and a deduced ...

Quantum dot-based molecular imaging of cancer cell growth using a clone formation assay.

Science.gov (United States)

Geng, Xia-Fei; Fang, Min; Liu, Shao-Ping; Li, Yan

2016-10-01

This aim of the present study was to investigate clonal growth behavior and analyze the proliferation characteristics of cancer cells. The MCF‑7 human breast cancer cell line, SW480 human colon cancer cell line and SGC7901 human gastric cancer cell line were selected to investigate the morphology of cell clones. Quantum dot‑based molecular targeted imaging techniques (which stained pan‑cytokeratin in the cytoplasm green and Ki67 in the cell nucleus yellow or red) were used to investigate the clone formation rate, cell morphology, discrete tendency, and Ki67 expression and distribution in clones. From the cell clone formation assay, the MCF‑7, SW480 and SGC7901 cells were observed to form clones on days 6, 8 and 12 of cell culture, respectively. These three types of cells had heterogeneous morphology, large nuclear:cytoplasmic ratios, and conspicuous pathological mitotic features. The cells at the clone periphery formed multiple pseudopodium. In certain clones, cancer cells at the borderline were separated from the central cell clusters or presented a discrete tendency. With quantum dot‑based molecular targeted imaging techniques, cells with strong Ki67 expression were predominantly shown to be distributed at the clone periphery, or concentrated on one side of the clones. In conclusion, cancer cell clones showed asymmetric growth behavior, and Ki67 was widely expressed in clones of these three cell lines, with strong expression around the clones, or aggregated at one side. Cell clone formation assay based on quantum dots molecular imaging offered a novel method to study the proliferative features of cancer cells, thus providing a further insight into tumor biology.

The first isolation and molecular characterization of Toxoplasma gondii from horses in Serbia.

NARCIS (Netherlands)

Klun, Ivana; Uzelac, Aleksandra; Villena, Isabelle; Mercier, Aurélien; Bobić, Branko; Nikolić, Aleksandra; Rajnpreht, Irena; Opsteegh, Marieke; Aubert, Dominique; Blaga, Radu; van der Giessen, Joke; Djurković-Djaković, Olgica

2017-01-01

Consumption of undercooked or insufficiently cured meat is a major risk factor for human infection with Toxoplasma gondii. Although horsemeat is typically consumed rare or undercooked, information on the risk of T. gondii from infected horse meat to humans is scarce. Here, we present the results of

Molecular and serological prevalence of Toxoplasma gondii and Anaplasma spp. infection in goats from Chongqing Municipality, China

Directory of Open Access Journals (Sweden)

Zhou Zuoyong

2018-01-01

Full Text Available Toxoplasmosis and anaplasmosis are severe zoonotic diseases, the former caused by Toxoplasma gondii and the latter by Anaplasma spp. In the present study, 332 goat blood samples were randomly collected from Chongqing Municipality, China to screen for T. gondii and Anaplasma spp. We used a polymerase chain reaction (PCR to detect DNA, and enzyme-linked immunosorbent assay (ELISA to test for T. gondii antibodies. The prevalence of T. gondii and Anaplasma spp. was 38% and 35% respectively by PCR, and 42% for T. gondii antibodies by ELISA. The co-infection rate by T. gondii and Anaplasma was 13%, where the two predominant pathogens co-infecting were Anaplasma phagocytophilum + A. bovis (10%, followed by T. gondii + A. phagocytophilum (9.64%. While co-infection by three pathogens varied ranging from 1.81% to 5.72%, less than 1% of goats were found to be positive for four pathogens. This is the first investigation of T. gondii and Anaplasma spp. infection in goats from Chongqing.

Molecular cloning of cellulase genes from indigenous bacterial isolates

International Nuclear Information System (INIS)

Jong Bor Chyan; Pauline Liew Woan Ying; Mat Rasol Awang

2006-01-01

Indigenous cellulolytic bacterial isolates having high activities in degrading carboxymethyl cellulose (CMC) were isolated from local environments. Identification of these isolates were performed by molecular techniques. By using polymerase chain reaction (PCR) techniques, PCR products encoding cellulase gene were amplified from the total genomic DNAs. Purified PCR product was successfully cloned and expressed in Escherichia coli host system. The complete nucleotide sequences of the cellulase genes determined. The analysis of amino acid sequences deduced from the genes indicated that the cloned DNA fragments show high homology to those of endoglucanase genes of family GH5. All cloned genes consist of an N-terminal signal peptide, a catalytic domain of family 5 glycosyl hydrolase and a cellulose-binding domain of family III. (Author)

Molecular cloning and characterization of a novel human testis ...

Indian Academy of Sciences (India)

Molecular cloning and characterization of a novel human testis-specific gene by use of ... pared against 70 other libraries, and the hits showing >10- fold differences .... proteins or testis-development-related proteins such as TSP-. NY, TPX1 ...

Molecular cloning and characterization of a putative OGG_N domain ...

African Journals Online (AJOL)

Molecular cloning and characterization of a putative OGG_N domain from the camel, Camelus dromedarius. Farid Shokry Ataya, Mohammad Saud Alanazi, Dalia Fouad, Hehsam Mahmoud Saeed, Mohammad Bazzi ...

First molecular evidence of Toxoplasma gondii in opossums (Didelphis virginiana) from Yucatan, Mexico

OpenAIRE

Torres-Castro, M.; Noh-Pech, H.; Puerto-Hern?ndez, R.; Reyes-Hern?ndez, B.; Panti-May, A.; Hern?ndez-Betancourt, S.; Yeh-Gorocica, A.; Gonz?lez-Herrera, L.; Zavala-Castro, J.; Puerto, F.I.

2016-01-01

Toxoplasma gondii is an obligate intracellular parasite recognized as a causal agent of toxoplasmosis; zoonotic disease endemic in many countries worldwide, including Mexico. Different species of animals participate in the wild cycle infection, including opossums of the species Didelphis virginiana. Thirteen D. virginiana were captured in Yucatan, Mexico. Detection of T. gondii was achieved by Polymerase Chain Reaction, which determined an infection of 76.9% (10/13) in brains. Positive amplic...

Structure of Toxoplasma gondii fructose-1,6-bisphosphate aldolase

International Nuclear Information System (INIS)

Boucher, Lauren E.; Bosch, Jürgen

2014-01-01

The structure of T. gondii fructose-1,6-bisphosphate aldolase, a glycolytic enzyme and structural component of the invasion machinery, was determined to a resolution of 2.0 Å. The apicomplexan parasite Toxoplasma gondii must invade host cells to continue its lifecycle. It invades different cell types using an actomyosin motor that is connected to extracellular adhesins via the bridging protein fructose-1,6-@@bisphosphate aldolase. During invasion, aldolase serves in the role of a structural bridging protein, as opposed to its normal enzymatic role in the glycolysis pathway. Crystal structures of the homologous Plasmodium falciparum fructose-1,6-bisphosphate aldolase have been described previously. Here, T. gondii fructose-1,6-bisphosphate aldolase has been crystallized in space group P22 1 2 1 , with the biologically relevant tetramer in the asymmetric unit, and the structure has been determined via molecular replacement to a resolution of 2.0 Å. An analysis of the quality of the model and of the differences between the four chains in the asymmetric unit and a comparison between the T. gondii and P. falciparum aldolase structures is presented

Taxonomy Icon Data: Toxoplasma gondii [Taxonomy Icon

Lifescience Database Archive (English)

Full Text Available Toxoplasma gondii Toxoplasma gondii Toxoplasma_gondii_L.png Toxoplasma_gondii_NL.png Toxoplasma..._gondii_S.png Toxoplasma_gondii_NS.png http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Toxoplasma...+gondii&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Toxoplasma+gondii&t=NL http://biosciencedbc.j...p/taxonomy_icon/icon.cgi?i=Toxoplasma+gondii&t=S http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Toxoplas...ma+gondii&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=113 ...

LILRB4 Decrease on uDCs Exacerbate Abnormal Pregnancy Outcomes Following Toxoplasma gondii Infection

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Shaowei Zhan

2018-03-01

Full Text Available Toxoplasma gondii (T. gondii infection in early pregnancy can result in miscarriage, dead fetus, and other abnormalities. The LILRB4 is a central inhibitory receptor in uterine dendritic cells (uDCs that plays essential immune-regulatory roles at the maternal–fetal interface. In this study, T. gondii-infected human primary uDCs and T. gondii-infected LILRB4-/- pregnant mice were utilized. The immune mechanisms underlying the role of LILRB4 on uDCs were explored in the development of abnormal pregnancy outcomes following T. gondii infection in vitro and in vivo. Our results showed that the expression levels of LILRB4 on uDCs from normal pregnant mice were obviously higher than non-pregnant mice, and peaked in mid-gestation. The LILRB4 expression on uDC subsets, especially tolerogenic subsets, from mid-gestation was obviously down-regulated after T. gondii infection and LILRB4 decrease could further regulate the expression of functional molecules (CD80, CD86, and HLA-DR or MHC II on uDCs, contributing to abnormal pregnancy outcomes. Our results will shed light on the molecular immune mechanisms of uDCs in abnormal pregnancy outcomes by T. gondii infection.

Detection of Toxoplasma gondii DNA in Brazilian oysters (Crassostrea rhizophorae).

Science.gov (United States)

Ribeiro, L A; Santos, L K N S S; Brito, P A; Maciel, B M; Da Silva, A V; Albuquerque, G R

2015-05-04

The aim of this study was to detect evidence of Toxoplasma gondii using polymerase chain reaction (PCR)-based techniques in oysters (Crassostrea rhizophorae) obtained from the southern coastal region of Bahia, Brazil. A total of 624 oysters were collected, and the gills and digestive glands were dissected. Each tissue sample was separated into pools containing tissues (of the same type) from three animals, leading to a total of 416 experimental samples for analysis (208 samples each from the gills and digestive glands). Molecular analysis using PCR-based detection of the T. gondii AF 146527 repetitive fragment yielded negative results for all samples. However, when nested-PCR was used for detection of the T. gondii SAG-1 gene, 17 samples were positive, with the gills being the tissue with maximal detection of the parasite. These positive results were confirmed by sample sequencing. It is therefore suggested that C. rhizophorae oysters are capable of filtering and retaining T. gondii oocysts in their tissue. This represents a risk to public health because they are traditionally ingested in natura.

Molecular Cloning Designer Simulator (MCDS): All-in-one molecular cloning and genetic engineering design, simulation and management software for complex synthetic biology and metabolic engineering projects.

Science.gov (United States)

Shi, Zhenyu; Vickers, Claudia E

2016-12-01

Molecular Cloning Designer Simulator (MCDS) is a powerful new all-in-one cloning and genetic engineering design, simulation and management software platform developed for complex synthetic biology and metabolic engineering projects. In addition to standard functions, it has a number of features that are either unique, or are not found in combination in any one software package: (1) it has a novel interactive flow-chart user interface for complex multi-step processes, allowing an integrated overview of the whole project; (2) it can perform a user-defined workflow of cloning steps in a single execution of the software; (3) it can handle multiple types of genetic recombineering, a technique that is rapidly replacing classical cloning for many applications; (4) it includes experimental information to conveniently guide wet lab work; and (5) it can store results and comments to allow the tracking and management of the whole project in one platform. MCDS is freely available from https://mcds.codeplex.com.

Toxoplasma gondii decreases the reproductive fitness in mice

Czech Academy of Sciences Publication Activity Database

Dvořáková-Hortová, K.; Šídlová, A.; Děd, Lukáš; Hladovcová, D.; Vieweg, M.; Weidner, W.; Steger, K.; Stopka, P.; Paradowska-Dogan, A.

2014-01-01

Roč. 9, č. 6 (2014), s. 1-11 E-ISSN 1932-6203 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:86652036 Keywords : Toxoplasma gondii * reproductive fitness * DNA methylation Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.234, year: 2014

Interaction and cystogenesis of Toxoplasma gondii within skeletal muscle cells in vitro

Directory of Open Access Journals (Sweden)

Erick Vaz Guimarães

2009-03-01

Full Text Available Infection by the protozoan parasite Toxoplasma gondii is widely prevalent in humans and animals. To prevent human infection, all meat should be well cooked before consumption, since the parasite is present in skeletal muscle. In this context, the use of skeletal muscle cells (SkMCs as a cellular model opens up new approaches to investigate T. gondii-host cell interactions. Immunofluorescent detection of proteins that are stage-specific for bradyzoites indicated that complete cystogenesis of T. gondii in in vitro cultures of SkMCs occurs after 96 h of infection. Ultrastructural analysis showed that, after 48 h of interaction, there were alterations on the parasitophorous vacuole membrane, including greater thickness and increased electron density at the inner face of the membrane. The present study demonstrates the potential use of primary cultures of SkMCs to evaluate different molecular aspects of T. gondii invasion and cystogenesis and presents a promising in vitro model for the screening of drug activities toward tissue cysts and bradyzoites.

Toxoplasma gondii in Blood Donors: A Study in Boyer-Ahmad County, Southwest Iran

Directory of Open Access Journals (Sweden)

Abdolali Moshfe

2018-01-01

Full Text Available Toxoplasma gondii is an important foodborne protozoan that can be transmitted through infected blood containing tachyzoite form of the parasite. The current study aimed to evaluate the prevalence of T. gondii infection and related risk factors among healthy blood donors in Boyer-Ahmad County, southwest Iran. Blood samples were taken from 285 healthy blood donors who voluntarily agreed to participate in this study. Sera and buffy coat were isolated from the blood samples for serological and molecular evaluations. The sera were tested for anti-T. gondii antibodies (both IgG and IgM, using a commercial ELISA kit. The buffy coat of seropositive cases was evaluated for detection of T. gondii DNA by PCR. Moreover, a structured questionnaire, containing socioepidemiological data and possible risk factors, was filled out by each participant during sample collection. Anti-T. gondii antibodies were detected in sera of 48/285 (16.8% participants. Only two of the subjects (0.7% were seropositive for both IgG and IgM antibodies. T. gondii DNA was not detected in buffy coat of any of the seropositive cases. Risk factors such as contact with soil (OR, 9.7; 95% CI, 4.9–19.4 and consumption of semicooked meat (OR, 2.5; 95% CI, 1.2–5.03 were statistically associated with seropositivity to T. gondii. The seroprevalence rate of T. gondii antibodies in the blood donors of Boyer-Ahmad County was not high in comparison with other regions in Iran. In this study, consumption of undercooked meats, job, and contact with soil were independent risk factors associated with T. gondii infection, which can be considered as potential sources of T. gondii infection.

Loop-mediated isothermal amplification (LAMP): Early detection of Toxoplasma gondii infection in mice

OpenAIRE

Kong, Qing-Ming; Lu, Shao-Hong; Tong, Qun-Bo; Lou, Di; Chen, Rui; Zheng, Bin; Kumagai, Takashi; Wen, Li-Yong; Ohta, Nobuo; Zhou, Xiao-Nong

2012-01-01

Abstract Background Toxoplasmosis is a widespread zoonotic parasitic disease that occurs in both animals and humans. Traditional molecular assays are often difficult to perform, especially for the early diagnosis of Toxoplasma gondii infections. Here, we established a novel loop-mediated isothermal amplification targeting the 529 bp repeat element (529 bp-LAMP) to detect T. gondii DNA in blood samples of experimental mice infected with tachyzoites of the RH strain. Findings The assay was perf...

Molecular cloning of osteoma-inducing replication-competent murine leukemia viruses from the RFB osteoma virus stock

DEFF Research Database (Denmark)

Pedersen, Lene; Behnisch, Werner; Schmidt, Jörg

1992-01-01

We report the molecular cloning of two replication-competent osteoma-inducing murine leukemia viruses from the RFB osteoma virus stock (M. P. Finkel, C. A. Reilly, Jr., B. O. Biskis, and I. L. Greco, p. 353-366, in C. H. G. Price and F. G. M. Ross, ed., Bone--Certain Aspects of Neoplasia, 1973......). Like the original RFB osteoma virus stock, viruses derived from the molecular RFB clones induced multiple osteomas in mice of the CBA/Ca strain. The cloned RFB viruses were indistinguishable by restriction enzyme analysis and by nucleotide sequence analysis of their long-terminal-repeat regions...

Toxoplasma gondii and schizophrenia

OpenAIRE

Mokhtari Mohammadreza; Mokhtari Mojgan

2006-01-01

Recent epidemiologic studies indicate that infectious agents may contribute to some cases of schizophrenia. In animals, infection with Toxoplasma gondii can alter behavior and neurotransmitter function. In humans, acute infection with T. gondii can produce psychotic symptoms similar to those displayed by persons with schizophrenia. Since 1953, a total of 19 studies of T. gondii antibodies in persons with schizophrenia and other severe psychiatric disorders and in controls have been reported; ...

A universal mini-vector and an annealing of PCR products (APP)-based cloning strategy for convenient molecular biological manipulations.

Science.gov (United States)

Liu, Xia; Li, Tuoping; Hart, Darren J; Gao, Song; Wang, Hongling; Gao, Herui; Xu, Shumin; Zhang, Yifeng; Liu, Yifei; An, Yingfeng

2018-03-18

Currently, the most widely used strategies for molecular cloning are sticky-end ligation-based cloning, TA cloning, blunt-end ligation-based cloning and ligase-independent cloning. In this study we have developed a novel mini-vector pANY1 which can simultaneously meet the requirements of all these cloning strategies. In addition, the selection of appropriate restriction digestion sites is difficult in some cases because of the presence of internal sites. In this study, an annealing of PCR products (APP)-based sticky-end cloning strategy was introduced to avoid this issue. Additionally, false positives occur during molecular cloning, which increases the workload of isolating positive clones. The plasmid pANY1 contains a ccdB cassette between multiple cloning sites, which efficiently avoids these false positives. Therefore, this mini-vector should serve as a useful tool with wide applications in biosciences, agriculture, food technologies, etc. Copyright © 2018 Elsevier Inc. All rights reserved.

Molecular transformation, gene cloning, and gene expression systems for filamentous fungi

Science.gov (United States)

Gold, Scott E.; Duick, John W.; Redman, Regina S.; Rodriguez, Rusty J.

2001-01-01

This chapter discusses the molecular transformation, gene cloning, and gene expression systems for filamentous fungi. Molecular transformation involves the movement of discrete amounts of DNA into cells, the expression of genes on the transported DNA, and the sustainable replication of the transforming DNA. The ability to transform fungi is dependent on the stable replication and expression of genes located on the transforming DNA. Three phenomena observed in bacteria, that is, competence, plasmids, and restriction enzymes to facilitate cloning, were responsible for the development of molecular transformation in fungi. Initial transformation success with filamentous fungi, involving the complementation of auxotrophic mutants by exposure to sheared genomic DNA or RNA from wt isolates, occurred with low transformation efficiencies. In addition, it was difficult to retrieve complementing DNA fragments and isolate genes of interest. This prompted the development of transformation vectors and methods to increase efficiencies. The physiological studies performed with fungi indicated that the cell wall could be removed to generate protoplasts. It was evident that protoplasts could be transformed with significantly greater efficiencies than walled cells.

Molecular detection of Toxoplasma gondii and Neospora caninum in birds from South Africa.

Science.gov (United States)

Lukášová, Radka; Kobédová, Kateřina; Halajian, Ali; Bártová, Eva; Murat, Jean-Benjamin; Rampedi, Kgethedi Michael; Luus-Powell, Wilmien J

2018-02-01

There are not any records on the detection of Toxoplasma gondii and Neospora caninum in tissues of wild birds in the African continent. The aim of the study was to investigate the occurrence of DNA from these protozoan parasites in brain tissue samples collected in years 2014-2015 from 110 wild and domestic birds of 15 orders. Birds came mainly from the province of Limpopo (n=103); the other seven birds came from other five provinces of South Africa. Parasite DNAs were detected by PCR in animal brains. While all samples were negative for N. caninum, T. gondii DNA was detected in three (2.7%) birds: a Red-eyed Dove (Streptopelia semitorquata), a Laughing Dove (S. senegalensis) and a Southern-Yellow-billed Hornbill (Tockus leucomelas), all from Limpopo province. Positive samples were selected for genotyping by a 15 microsatellite markers method in a single multiplex PCR assay. Only the sample from the Red-eyed Dove was successfully genotyped and characterized as type II. This is the first detection of T. gondii in tissue of native African wild birds and the first study focusing on N. caninum in birds from South Africa. Copyright © 2017. Published by Elsevier B.V.

Particle infectivity of HIV-1 full-length genome infectious molecular clones in a subtype C heterosexual transmission pair following high fidelity amplification and unbiased cloning

Energy Technology Data Exchange (ETDEWEB)

Deymier, Martin J., E-mail: mdeymie@emory.edu [Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road NE, Atlanta, GA 30329 (United States); Claiborne, Daniel T., E-mail: dclaibo@emory.edu [Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road NE, Atlanta, GA 30329 (United States); Ende, Zachary, E-mail: zende@emory.edu [Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road NE, Atlanta, GA 30329 (United States); Ratner, Hannah K., E-mail: hannah.ratner@emory.edu [Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road NE, Atlanta, GA 30329 (United States); Kilembe, William, E-mail: wkilembe@rzhrg-mail.org [Zambia-Emory HIV Research Project (ZEHRP), B22/737 Mwembelelo, Emmasdale Post Net 412, P/BagE891, Lusaka (Zambia); Allen, Susan, E-mail: sallen5@emory.edu [Zambia-Emory HIV Research Project (ZEHRP), B22/737 Mwembelelo, Emmasdale Post Net 412, P/BagE891, Lusaka (Zambia); Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA (United States); Hunter, Eric, E-mail: eric.hunter2@emory.edu [Emory Vaccine Center, Yerkes National Primate Research Center, 954 Gatewood Road NE, Atlanta, GA 30329 (United States); Department of Pathology and Laboratory Medicine, Emory University, Atlanta, GA (United States)

2014-11-15

The high genetic diversity of HIV-1 impedes high throughput, large-scale sequencing and full-length genome cloning by common restriction enzyme based methods. Applying novel methods that employ a high-fidelity polymerase for amplification and an unbiased fusion-based cloning strategy, we have generated several HIV-1 full-length genome infectious molecular clones from an epidemiologically linked transmission pair. These clones represent the transmitted/founder virus and phylogenetically diverse non-transmitted variants from the chronically infected individual's diverse quasispecies near the time of transmission. We demonstrate that, using this approach, PCR-induced mutations in full-length clones derived from their cognate single genome amplicons are rare. Furthermore, all eight non-transmitted genomes tested produced functional virus with a range of infectivities, belying the previous assumption that a majority of circulating viruses in chronic HIV-1 infection are defective. Thus, these methods provide important tools to update protocols in molecular biology that can be universally applied to the study of human viral pathogens. - Highlights: • Our novel methodology demonstrates accurate amplification and cloning of full-length HIV-1 genomes. • A majority of plasma derived HIV variants from a chronically infected individual are infectious. • The transmitted/founder was more infectious than the majority of the variants from the chronically infected donor.

Particle infectivity of HIV-1 full-length genome infectious molecular clones in a subtype C heterosexual transmission pair following high fidelity amplification and unbiased cloning

International Nuclear Information System (INIS)

Deymier, Martin J.; Claiborne, Daniel T.; Ende, Zachary; Ratner, Hannah K.; Kilembe, William; Allen, Susan; Hunter, Eric

2014-01-01

The high genetic diversity of HIV-1 impedes high throughput, large-scale sequencing and full-length genome cloning by common restriction enzyme based methods. Applying novel methods that employ a high-fidelity polymerase for amplification and an unbiased fusion-based cloning strategy, we have generated several HIV-1 full-length genome infectious molecular clones from an epidemiologically linked transmission pair. These clones represent the transmitted/founder virus and phylogenetically diverse non-transmitted variants from the chronically infected individual's diverse quasispecies near the time of transmission. We demonstrate that, using this approach, PCR-induced mutations in full-length clones derived from their cognate single genome amplicons are rare. Furthermore, all eight non-transmitted genomes tested produced functional virus with a range of infectivities, belying the previous assumption that a majority of circulating viruses in chronic HIV-1 infection are defective. Thus, these methods provide important tools to update protocols in molecular biology that can be universally applied to the study of human viral pathogens. - Highlights: • Our novel methodology demonstrates accurate amplification and cloning of full-length HIV-1 genomes. • A majority of plasma derived HIV variants from a chronically infected individual are infectious. • The transmitted/founder was more infectious than the majority of the variants from the chronically infected donor

Enrofloxacin and Toltrazuril Are Able to Reduce Toxoplasma gondii Growth in Human BeWo Trophoblastic Cells and Villous Explants from Human Third Trimester Pregnancy

Directory of Open Access Journals (Sweden)

Rafaela J. da Silva

2017-07-01

Full Text Available Classical treatment for congenital toxoplasmosis is based on combination of sulfadiazine and pyrimethamine plus folinic acid. Due to teratogenic effects and bone marrow suppression caused by pyrimethamine, the establishment of new therapeutic strategies is indispensable to minimize the side effects and improve the control of infection. Previous studies demonstrated that enrofloxacin and toltrazuril reduced the incidence of Neospora caninum and Toxoplasma gondii infection. The aim of the present study was to evaluate the efficacy of enrofloxacin and toltrazuril in the control of T. gondii infection in human trophoblast cells (BeWo line and in human villous explants from the third trimester. BeWo cells and villous were treated with several concentrations of enrofloxacin, toltrazuril, sulfadiazine, pyrimethamine, or combination of sulfadiazine+pyrimethamine, and the cellular or tissue viability was verified. Next, BeWo cells were infected by T. gondii (2F1 clone or the ME49 strain, whereas villous samples were only infected by the 2F1 clone. Then, infected cells and villous were treated with all antibiotics and the T. gondii intracellular proliferation as well as the cytokine production were analyzed. Finally, we evaluated the direct effect of enrofloxacin and toltrazuril in tachyzoites to verify possible changes in parasite structure. Enrofloxacin and toltrazuril did not decrease the viability of cells and villous in lower concentrations. Both drugs were able to significantly reduce the parasite intracellular proliferation in BeWo cells and villous explants when compared to untreated conditions. Regardless of the T. gondii strain, BeWo cells infected and treated with enrofloxacin or toltrazuril induced high levels of IL-6 and MIF. In villous explants, enrofloxacin induced high MIF production. Finally, the drugs increased the number of unviable parasites and triggered damage to tachyzoite structure. Taken together, it can be concluded that

Enrofloxacin and Toltrazuril Are Able to Reduce Toxoplasma gondii Growth in Human BeWo Trophoblastic Cells and Villous Explants from Human Third Trimester Pregnancy.

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da Silva, Rafaela J; Gomes, Angelica O; Franco, Priscila S; Pereira, Ariane S; Milian, Iliana C B; Ribeiro, Mayara; Fiorenzani, Paolo; Dos Santos, Maria C; Mineo, José R; da Silva, Neide M; Ferro, Eloisa A V; de Freitas Barbosa, Bellisa

2017-01-01

Classical treatment for congenital toxoplasmosis is based on combination of sulfadiazine and pyrimethamine plus folinic acid. Due to teratogenic effects and bone marrow suppression caused by pyrimethamine, the establishment of new therapeutic strategies is indispensable to minimize the side effects and improve the control of infection. Previous studies demonstrated that enrofloxacin and toltrazuril reduced the incidence of Neospora caninum and Toxoplasma gondii infection. The aim of the present study was to evaluate the efficacy of enrofloxacin and toltrazuril in the control of T. gondii infection in human trophoblast cells (BeWo line) and in human villous explants from the third trimester. BeWo cells and villous were treated with several concentrations of enrofloxacin, toltrazuril, sulfadiazine, pyrimethamine, or combination of sulfadiazine+pyrimethamine, and the cellular or tissue viability was verified. Next, BeWo cells were infected by T. gondii (2F1 clone or the ME49 strain), whereas villous samples were only infected by the 2F1 clone. Then, infected cells and villous were treated with all antibiotics and the T. gondii intracellular proliferation as well as the cytokine production were analyzed. Finally, we evaluated the direct effect of enrofloxacin and toltrazuril in tachyzoites to verify possible changes in parasite structure. Enrofloxacin and toltrazuril did not decrease the viability of cells and villous in lower concentrations. Both drugs were able to significantly reduce the parasite intracellular proliferation in BeWo cells and villous explants when compared to untreated conditions. Regardless of the T. gondii strain, BeWo cells infected and treated with enrofloxacin or toltrazuril induced high levels of IL-6 and MIF. In villous explants, enrofloxacin induced high MIF production. Finally, the drugs increased the number of unviable parasites and triggered damage to tachyzoite structure. Taken together, it can be concluded that enrofloxacin and

Gene Transfer and Molecular Cloning of the Human NGF Receptor

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Chao, Moses V.; Bothwell, Mark A.; Ross, Alonzo H.; Koprowski, Hilary; Lanahan, Anthony A.; Buck, C. Randall; Sehgal, Amita

1986-04-01

Nerve growth factor (NGF) and its receptor are important in the development of cells derived from the neural crest. Mouse L cell transformants have been generated that stably express the human NGF receptor gene transfer with total human DNA. Affinity cross-linking, metabolic labeling and immunoprecipitation, and equilibrium binding with 125I-labeled NGF revealed that this NGF receptor had the same size and binding characteristics as the receptor from human melanoma cells and rat PC12 cells. The sequences encoding the NGF receptor were molecularly cloned using the human Alu repetitive sequence as a probe. A cosmid clone that contained the human NGF receptor gene allowed efficient transfection and expression of the receptor.

Toxoplasma gondii in small neotropical wild felids

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William Alberto Cañon-Franco

2013-02-01

Full Text Available In the last decade, studies on wildlife worldwide have discovered key epidemiological aspects of the sylvatic cycle of Toxoplasma gondii. However, despite the known role of wild felines as definitive hosts in the transmission and maintenance of this parasite, few studies have focused on the involvement of these animals. Brazil exhibits the largest number of wild felid species in the Americas, all of which have a critical conservation status. However, serological detections, epidemiological studies and some molecular characterizations of T. gondii have primarily used Neotropical felid populations that are maintained in captivity, which does not reflect the disease behavior in free-living conditions. A systematic review of the worldwide scientific literature was conducted focusing on toxoplasmosis in small Neotropical felids. This review covered a number of aspects, including the state of scientific research, parasite transmission in the wild, the genetic characteristics of isolates, the relationship between these genetic characteristics and the pathogenicity of the parasite, and the risk factors linked to conflicts with humans. The present review shows the relevance of studying these felid populations based on their frequent interactions with humans in peri-urban areas and the need for further comprehensive studies to establish the real significance of T. gondii in public and animal health in tropical and temperate regions.

Dual congenital transmission of Toxoplasma gondii and Sarcocystis neurona in a late-term aborted pup from a chronically infected southern sea otter (Enhydra lutris nereis).

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Shapiro, Karen; Miller, Melissa A; Packham, Andrea E; Aguilar, Beatriz; Conrad, Patricia A; Vanwormer, Elizabeth; Murray, Michael J

2016-03-01

Toxoplasma gondii and Sarcocystis neurona are protozoan parasites with terrestrial definitive hosts, and both pathogens can cause fatal disease in a wide range of marine animals. Close monitoring of threatened southern sea otters (Enhydra lutris nereis) in California allowed for the diagnosis of dual transplacental transmission of T. gondii and S. neurona in a wild female otter that was chronically infected with both parasites. Congenital infection resulted in late-term abortion due to disseminated toxoplasmosis. Toxoplasma gondii and S. neurona DNA was amplified from placental tissue culture, as well as from fetal lung tissue. Molecular characterization of T. gondii revealed a Type X genotype in isolates derived from placenta and fetal brain, as well as in all tested fetal organs (brain, lung, spleen, liver and thymus). This report provides the first evidence for transplacental transmission of T. gondii in a chronically infected wild sea otter, and the first molecular and immunohistochemical confirmation of concurrent transplacental transmission of T. gondii and S. neurona in any species. Repeated fetal and/or neonatal losses in the sea otter dam also suggested that T. gondii has the potential to reduce fecundity in chronically infected marine mammals through parasite recrudescence and repeated fetal infection.

HLA-DQBl*0402 alleles polymorphisms detected in Javanese HIV patients with positive anti-Toxoplasma gondii IgM

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Sari, Yulia; Haryati, Sri; Prasetyo, Afiono Agung; Hartono, Adnan, Zainal Arifin

2017-02-01

The human leukocyte antigen (HLA)-DQB1 gene polymorphisms may associated with the infection risk of Toxoplasma gondii in HIV patients. The HLA-DQB1*0402 in HIV-1-positive patients could be considered risk factors for developing neurological opportunistic infections, mainly Toxoplasma encephalitis. However, the HLA-DQB1*0402 gene polymorphisms status in the Javanese HIV patients is unknown. This study evaluated the prevalence of HLA-DQB*0402 alleles polymorphisms in Javanese HIV patients with positive anti-Toxoplasma gondii IgM status. Since 2009 our research group performing a molecular epidemiology of blood borne viruses in Central Java Indonesia, by collecting the epidemiological and clinical data from the high risk communities. All blood samples were screened for blood borne pathogens by serological and molecular assays including for HIV and Toxoplasma gondii. The genomic DNA was isolated from the whole blood samples. Genetic polymorphisms of HLA-DQB1*0402 alleles were detected with polymerase chain reaction-sequence-specific primers (PCR-SSPs) technique. The genotypes were defined according to generated fragment patterns in the agarose gel electrophoresis analysis of PCR products. All of the samples were tested at least in duplicate. HLA-DQB1*0402 alleles were detected in 20.8% (16/77) patients and not detected in all HIV positive samples with negative anti-Toxoplasma gondii IgM status (n= 200). The HLA-DQB1*0402 alleles polymorphisms were detected in Javanese HIV patients with positive anti-Toxoplasma gondii IgM. The polymorphisms found may have association with the infection risk of Toxoplasma gondii in HIV patients.

Molecular cloning of a peptidylglycine alpha-hydroxylating monooxygenase from sea anemones

DEFF Research Database (Denmark)

Hauser, F; Williamson, M; Grimmelikhuijzen, C J

1997-01-01

conserved regions of PHM, we have now cloned a PHM from the sea anemone Calliactis parasitica showing 42% amino acid sequence identity with rat PHM. Among the conserved (identical) amino acid residues are five histidine and one methionine residue, which bind two Cu2+ atoms that are essential for PHM...... activity. No cDNA coding for PAL could be identified, suggesting that sea anemone PAL is coded for by a gene that is different from the sea anemone PHM gene, a situation similar to the one found in insects. This is the first report on the molecular cloning of a cnidarian PHM. Udgivelsesdato: 1997-Dec-18...

Seroprevalence of Toxoplasma gondii Infection in Patients of Intensive Care Unit in China: A Hospital Based Study

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Yong-Biao Zhang

2015-01-01

Full Text Available The objective of this study was to estimate the seroprevalence of Toxoplasma gondii infection in 394 patients of intensive care unit (ICU in a hospital between April 2010 and March 2012 and analyze the association between T. gondii infection and ICU patients according to the species of disease. Toxoplasma serology was evaluated by ELISA method using a commercially available kit. Data of patients were obtained from the patients, informants, and medical examination records. Seventy-four (18.78% of 394 patients were positive for anti-T. gondii IgG antibodies demonstrating latent infection. Of these, the highest T. gondii seroprevalence was found in the age group of 31–45 years (27.45%, and the lowest was found in the age group of <30 years (12.5%. In addition, females (21.6% had a higher seroprevalence than males (18.36%. With respect to the species of disease, the patients with kidney diseases (57.14%, lung diseases (27.84%, and brain diseases (24% had high T. gondii seroprevalence. The present study represents the first survey of T. gondii seroprevalence in ICU patients in China, revealing an 18.78% seropositivity. Considering the particularities of ICU patients, molecular identification, genetic characterization, and diagnosis of T. gondii should be considered in future study.

Toxoplasma gondii, Mental Health and Shizophrenia

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Sibel Cevizci

2013-04-01

Full Text Available Protecting and promoting of mental health is one of the major application areas of public health. In particular, Toxoplasma gondii, which is a protozoal zoonosis common in Turkey, it is closely related to veterinary public health. In recent years, T.gondii can induce behavioral changes, may play a role in schizophrenia as an etiologic factor. Results of the recently performed studies shows that T.gondii may be a potential factor for some neuropathological changes in brain and suicide attemption. The purpose of this review is to present the data on recent epidemiology of T.gondii, mental health effects (changes in behavior, suicide, etc., the relationship between T.gondii and schizophrenia and offer some recommendations for protecting of public health. [TAF Prev Med Bull 2013; 12(2.000: 199-208

The 5' untranslated region of a novel infectious molecular clone of the dicistrovirus cricket paralysis virus modulates infection.

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Kerr, Craig H; Wang, Qing S; Keatings, Kathleen; Khong, Anthony; Allan, Douglas; Yip, Calvin K; Foster, Leonard J; Jan, Eric

2015-06-01

Dicistroviridae are a family of RNA viruses that possesses a single-stranded positive-sense RNA genome containing two distinct open reading frames (ORFs), each preceded by an internal ribosome entry site that drives translation of the viral structural and nonstructural proteins, respectively. The type species, Cricket paralysis virus (CrPV), has served as a model for studying host-virus interactions; however, investigations into the molecular mechanisms of CrPV and other dicistroviruses have been limited as an established infectious clone was elusive. Here, we report the construction of an infectious molecular clone of CrPV. Transfection of in vitro-transcribed RNA from the CrPV clone into Drosophila Schneider line 2 (S2) cells resulted in cytopathic effects, viral RNA accumulation, detection of negative-sense viral RNA, and expression of viral proteins. Transmission electron microscopy, viral titers, and immunofluorescence-coupled transwell assays demonstrated that infectious viral particles are released from transfected cells. In contrast, mutant clones containing stop codons in either ORF decreased virus infectivity. Injection of adult Drosophila flies with virus derived from CrPV clones but not UV-inactivated clones resulted in mortality. Molecular analysis of the CrPV clone revealed a 196-nucleotide duplication within its 5' untranslated region (UTR) that stimulated translation of reporter constructs. In cells infected with the CrPV clone, the duplication inhibited viral infectivity yet did not affect viral translation or RNA accumulation, suggesting an effect on viral packaging or entry. The generation of the CrPV infectious clone provides a powerful tool for investigating the viral life cycle and pathogenesis of dicistroviruses and may further understanding of fundamental host-virus interactions in insect cells. Dicistroviridae, which are RNA viruses that infect arthropods, have served as a model to gain insights into fundamental host-virus interactions in

Anti-Toxoplasma gondii antibodies in patients with beta-hemoglobinopathies: the first report in the Americas.

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Ferreira, Marina Neves; Bonini-Domingos, Claudia Regina; Fonseca Estevão, Isabeth; de Castro Lobo, Clarice Lopes; Souza Carrocini, Gisele Cristina; Silveira-Carvalho, Aparecida Perpétuo; Ricci, Octávio; de Mattos, Luiz Carlos; Brandão de Mattos, Cinara Cássia

2017-06-14

In Brazil, there have been no previous studies of Toxoplasma gondii infection in sickle cell anemia patients and carriers of severe forms of beta-thalassemia. This study evaluated T. gondii infection in patients with beta-hemoglobinopathies. A total of 158 samples, 77 (48.7%) men and 81 (51.3%) women, were evaluated. Three groups were formed: G1 (85 patients with sickle cell disease); G2 (11 patients with homozygous beta-thalassemia; G3 (62 patients with heterozygous beta-thalassemia). ELISA was employed to identify anti-T. gondii IgM and IgG antibodies, and molecular analysis was performed to determine beta-hemoglobin mutations. Fisher's exact test was used to compare frequencies of anti-T. gondii IgM and IgG antibodies in respect to gender and age. Anti-T. gondii IgG antibodies were found in 43.5% of individuals in G1, 18.1% in G2 and 50% in G3. All samples from G1 and G2 were seronegative for anti-T. gondii IgM antibodies, but 3.2% from G3 were seropositive. Considering anti-T. gondii IgG antibodies, no statistical significant differences were found between these groups nor in seroprevalence between genders within each group. Despite this, comparisons of the mean ages between G1, G2 and G3 were statistically significant (G2 vs. G1: p value = 0.0001; G3 vs. G1: p-value <0.0001; G3 vs. G2: p-value = 0.0001). A comparison by age of patients with sickle cell anemia showed a trend of lower risk of infection among younger individuals. Therefore, this study demonstrates that T. gondii infection occurs in patients with beta-thalassemia and sickle cell anemia in Brazil as seen by the presence of anti-T. gondii IgM and IgG antibodies.

Identification of molecular performance from oil palm clones based on SSR markers

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Putri, Lollie Agustina P.; Basyuni, M.; Bayu, Eva S.; Arvita, D.; Arifiyanto, D.; Syahputra, I.

2018-03-01

In Indonesia, the oil palms are an important economic crop, producing food and raw materials for the food, confectionary, cosmetics and oleo-chemical industrial demands of oil palm products. Clonal oil palm offers the potential for greater productivity because it is possible to establish uniform tree stands comprising identical copies (clones) of a limited number of highly productive oil palms. Unfortunately, tissue culture sometimes accentuates the expression of detects in oil palm, particularly when embryogenesis is induced in particullar callus for prolonged periods. This research is conducted by taking individual tree sample of clone germplasm two years old. The purpose of this research is to molecular performance analysis of some oil palm clones based on SSR markers. A total of 30 trees oil palm clones were used for analysis. In this experiment, the DNA profile diversity was assessed using five loci of oil palm’s specific SSR markers. The results of the experiment indicated out of 3 SSR markers (FR-0779, FR-3663 and FR-0782) showed monomorphic of PCR product and 2 SSR markers (FR-0783 and FR- 3745) showed polymorphic of PCR product. There are 10 total number of PCR product. These preliminary results demonstrated SSR marker can be used to evaluate genetic relatedness among trees of oil palm clones.

Molecular cloning and characterization of arginine kinase gene of Toxocara canis.

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Sahu, Shivani; Samanta, S; Harish, D R; Sudhakar, N R; Raina, O K; Shantaveer, S B; Madhu, D N; Kumar, Ashok

2015-06-01

Toxocara canis is an important gastrointestinal nematode of dogs and also a causative agent of visceral larva migrans in humans. Arginine kinase (AK) gene is one of the important biomolecule of phosphagen kinase of T. canis which is emerging as an exciting novel diagnostic target in toxocarosis. The present study was carried out to clone and characterize AK gene of T. canis for future utilization as a diagnostic molecule. Total RNA was extracted from intact adult worms and reverse transcription was done with oligo dT primers to obtain complementary DNA (cDNA). Polymerase chain reaction (PCR) was carried out using cDNA as template with specific primers which amplified a product of 1,202 bp. The amplicon was cloned into pDrive cloning vector and clone was confirmed by colony PCR and restriction endonuclease analysis. Sequence analysis of the gene showed 99.8 and 77.9 % homology with the published AK gene of T. canis (EF015466.1) and Ascaris suum respectively. Structural analysis shown that the mature AK protein consist of 400 amino acids with a molecular wt of 45360.73 Da. Further expression studies are required for producing the recombinant protein for its evaluation in the diagnosis of T. canis infection in humans as well as in adult dogs.

Using molecular epidemiology to track Toxoplasma gondii from terrestrial carnivores to marine hosts: implications for public health and conservation.

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VanWormer, Elizabeth; Miller, Melissa A; Conrad, Patricia A; Grigg, Michael E; Rejmanek, Daniel; Carpenter, Tim E; Mazet, Jonna A K

2014-01-01

Environmental transmission of the zoonotic parasite Toxoplasma gondii, which is shed only by felids, poses risks to human and animal health in temperate and tropical ecosystems. Atypical T. gondii genotypes have been linked to severe disease in people and the threatened population of California sea otters. To investigate land-to-sea parasite transmission, we screened 373 carnivores (feral domestic cats, mountain lions, bobcats, foxes, and coyotes) for T. gondii infection and examined the distribution of genotypes in 85 infected animals sampled near the sea otter range. Nested PCR-RFLP analyses and direct DNA sequencing at six independent polymorphic genetic loci (B1, SAG1, SAG3, GRA6, L358, and Apico) were used to characterize T. gondii strains in infected animals. Strains consistent with Type X, a novel genotype previously identified in over 70% of infected sea otters and four terrestrial wild carnivores along the California coast, were detected in all sampled species, including domestic cats. However, odds of Type X infection were 14 times higher (95% CI: 1.3-148.6) for wild felids than feral domestic cats. Type X infection was also linked to undeveloped lands (OR = 22, 95% CI: 2.3-250.7). A spatial cluster of terrestrial Type II infection (P = 0.04) was identified in developed lands bordering an area of increased risk for sea otter Type II infection. Two spatial clusters of animals infected with strains consistent with Type X (P ≤ 0.01) were detected in less developed landscapes. Differences in T. gondii genotype prevalence among domestic and wild felids, as well as the spatial distribution of genotypes, suggest co-existing domestic and wild T. gondii transmission cycles that likely overlap at the interface of developed and undeveloped lands. Anthropogenic development driving contact between these cycles may increase atypical T. gondii genotypes in domestic cats and facilitate transmission of potentially more pathogenic genotypes to humans, domestic animals

Using molecular epidemiology to track Toxoplasma gondii from terrestrial carnivores to marine hosts: implications for public health and conservation.

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Elizabeth VanWormer

Full Text Available Environmental transmission of the zoonotic parasite Toxoplasma gondii, which is shed only by felids, poses risks to human and animal health in temperate and tropical ecosystems. Atypical T. gondii genotypes have been linked to severe disease in people and the threatened population of California sea otters. To investigate land-to-sea parasite transmission, we screened 373 carnivores (feral domestic cats, mountain lions, bobcats, foxes, and coyotes for T. gondii infection and examined the distribution of genotypes in 85 infected animals sampled near the sea otter range.Nested PCR-RFLP analyses and direct DNA sequencing at six independent polymorphic genetic loci (B1, SAG1, SAG3, GRA6, L358, and Apico were used to characterize T. gondii strains in infected animals. Strains consistent with Type X, a novel genotype previously identified in over 70% of infected sea otters and four terrestrial wild carnivores along the California coast, were detected in all sampled species, including domestic cats. However, odds of Type X infection were 14 times higher (95% CI: 1.3-148.6 for wild felids than feral domestic cats. Type X infection was also linked to undeveloped lands (OR = 22, 95% CI: 2.3-250.7. A spatial cluster of terrestrial Type II infection (P = 0.04 was identified in developed lands bordering an area of increased risk for sea otter Type II infection. Two spatial clusters of animals infected with strains consistent with Type X (P ≤ 0.01 were detected in less developed landscapes.Differences in T. gondii genotype prevalence among domestic and wild felids, as well as the spatial distribution of genotypes, suggest co-existing domestic and wild T. gondii transmission cycles that likely overlap at the interface of developed and undeveloped lands. Anthropogenic development driving contact between these cycles may increase atypical T. gondii genotypes in domestic cats and facilitate transmission of potentially more pathogenic genotypes to humans

Isolation of viable Toxoplasma gondii, molecular characterization, and seroprevalence in elk (Cervus canadensis) in Pennsylvania, USA.

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Dubey, J P; Brown, J; Verma, S K; Cerqueira-Cézar, C K; Banfield, J; Kwok, O C H; Ying, Y; Murata, F H A; Pradhan, A K; Su, C

2017-08-30

Toxoplasmosis is a worldwide zoonosis. The ingestion of uncooked/undercooked meat and consumption of water contaminated with Toxoplasma gondii oocysts excreted by felids are the main modes of transmission of this parasite. T. gondii has been reported in multiple cervid species; however, little is known of the parasite in North American elk (Cervus canadensis). In the present study, antibodies to T. gondii were detected in serum of wild elk from Pennsylvania collected during 2013-2016 by the modified agglutination test (MAT, cut-off 1:25); 221 of 317 (69.7%) had MAT titers of 1:25 in 19, 1:50 in 28, 1:100 in 34, and 1:200 or higher in 140. Thus most (44.1%) elk had relatively high titers. Seroprevalence was slightly higher in males (76.9%) than females (67.5%, not statistically significant, Chi-square tests, P<0.0001) and was higher in adults (76.5%) than yearlings (46.4%, Odds ratio 3.82; 95% CL 1.72-8.47; P=0.001) or calves (21.7%, Odds ratio 12.58; 95% CL 4.51-35.10; P<0.0001). Annual seroprevalence was relatively stable throughout the period tested and ranged from 66.6% to 72.2%. Of the 101 elk harvested in 2016, hearts were bioassayed from 20 elk and tongues were bioassayed from 56; all tongue samples were negative. Viable T. gondii was isolated from hearts of two female elk, one of these was a seronegative adult and the other was a calf with no serum available for testing. Both T. gondii isolates were cultivated in cell culture and DNA derived from tachyzoites was characterized using the PCR-RFLP markers including SAG1, SAG2 (5'- 3'SAG2 and altSAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. One isolate belongs to ToxoDB PCR-RFLP genotype #2 and the other is genotype #5. Both genotypes are frequently identified in animals in North America. Copyright © 2017. Published by Elsevier B.V.

Veterinary vaccines against Toxoplasma gondii

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Elisabeth A Innes

2009-03-01

Full Text Available Toxoplasma gondii has a very wide intermediate host range and is thought to be able to infect all warm blooded animals. The parasite causes a spectrum of different diseases and clinical symptoms within the intermediate hosts and following infection most animals develop adaptive humoral and cell-mediated immune responses. The development of protective immunity to T. gondii following natural infection in many host species has led researchers to look at vaccination as a strategy to control disease, parasite multiplication and establishment in animal hosts. A range of different veterinary vaccines are required to help control T. gondii infection which include vaccines to prevent congenital toxoplasmosis, reduce or eliminate tissue cysts in meat producing animals and to prevent oocyst shedding in cats. In this paper we will discuss some of the history, challenges and progress in the development of veterinary vaccines against T. gondii.

Immersing Undergraduate Students in the Research Experience: A Practical Laboratory Module on Molecular Cloning of Microbial Genes

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Wang, Jack T. H.; Schembri, Mark A.; Ramakrishna, Mathitha; Sagulenko, Evgeny; Fuerst, John A.

2012-01-01

Molecular cloning skills are an essential component of biological research, yet students often do not receive this training during their undergraduate studies. This can be attributed to the complexities of the cloning process, which may require many weeks of progressive design and experimentation. To address this issue, we incorporated an…

Seroprevalence of Toxoplasma gondii in Western Romania.

Science.gov (United States)

Olariu, Tudor Rares; Petrescu, Cristina; Darabus, Gheorghe; Lighezan, Rodica; Mazilu, Octavian

2015-08-01

Toxoplasma gondii is an obligate intracellular parasite that most commonly causes asymptomatic infection in immunocompetent hosts, but can have devastating consequences in congenitally infected infants and immunocompromised patients. We evaluated the seroprevalence of T. gondii in the general population in Western Romania. Sera from 304 individuals were analysed with the Pastorex Toxo test, which allows the simultaneous detection of T. gondii IgG and/or IgM antibodies. T. gondii antibodies were demonstrated in 197 individuals (64.8%) and the prevalence increased with age: 35.0% in those Romania.

Viability and infectivity analysis of Toxoplasma gondii under axenic conditions = Análise da viabilidade e infectividade de taquizoítas de Toxoplasma gondii em condições axênicas

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Ferreira-da-Silva, Marialice Fonseca

2010-01-01

Conclusões: nossos dados abrem novas perspectivas para o estudo de diferentes aspectos da biologia do Toxoplasma gondii, incluindo mecanismos de nutrição, ensaios experimentais de drogas in vitro, ou estudos da biologia celular e molecular diretamente sobre o parasito

Molecular cloning and characterization of l-methionine γ-lyase from Streptomyces avermitilis.

Science.gov (United States)

Kudou, Daizou; Yasuda, Eri; Hirai, Yoshiyuki; Tamura, Takashi; Inagaki, Kenji

2015-10-01

A pyridoxal 5'-phosphate-dependent methionine γ-lyase (MGL) was cloned from Streptomyces avermitilis catalyzed the degradation of methionine to α-ketobutyrate, methanethiol, and ammonia. The sav7062 gene (1,242 bp) was corresponded to 413 amino acid residues with a molecular mass of 42,994 Da. The deduced amino acid sequence showed a high degree of similarity to those of other MGL enzymes. The sav7062 gene was overexpressed in Escherichia coli. The enzyme was purified to homogeneity and exhibited the MGL catalytic activities. We cloned the enzyme that has the MGL activity in Streptomyces for the first time. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Seroprevalence of Toxoplasma gondii infection in Norwegian dairy goats

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Stormoen Marit

2012-12-01

Full Text Available Abstract Background Toxoplasma gondii is a major problem for the sheep industry as it may cause reproduction problems. The importance of T. gondii in Norwegian goat herds is uncertain, but outbreaks of toxoplasmosis in dairy goat farms have been recorded. The aim of this study was to describe the prevalence of T. gondii infection in Norwegian dairy goats by using serology. Findings Goat serum originally collected as part of two nationwide surveillance and control programmes between 2002 and 2008 were examined for T. gondii antibodies by using direct agglutination test. In total, 55 of 73 herds (75% had one or more serologically positive animals, while 377 of 2188 (17% of the individual samples tested positive for T. gondii antibodies. Conclusions This is the first prevalence study of T. gondii infection in Norwegian goats. The results show that Norwegian goat herds are commonly exposed to T. gondii. Nevertheless, the majority of goat herds have a low prevalence of antibody positive animals, which make them vulnerable to infections with T. gondii during the gestation period.

Molecular passportization of clones of karelian birch using PCRwith semi-specific primers

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Tatyana V Matveeva

2008-09-01

Full Text Available Using 4 clones of Karelian birch (Betula pendula Roth var caretica Merckl. from the collection of the karelian birch of the laboratory of genetics of Research Institute of Forest Genetics and Breeding, Voronezh, one normal tree of Betula pendula Roth, and one tree of B. pubescens Ehrh., collected from the nature, we have analyzed the possibility of application of PCR with semi-specific primers for molecular typing. We found primers with high percentage of polymorphic markers. These primers could be recommended for molecular typing of birch.

Toxoplasma gondii seroprevalence in breeding pigs in Estonia

DEFF Research Database (Denmark)

Santoro, Azzurra; Tagel, Maarja; Must, Kärt

2017-01-01

Background: Toxoplasma gondii is a widespread occurring parasite infecting warm-blooded animals, including pigs and humans. The aims of this study were to estimate the prevalence of anti-T. gondii antibodies and to evaluate risk factors for T. gondii seropositivity in breeding pigs raised in Esto...

First isolation and genotyping of Toxoplasma gondii from bats (Mammalia: Chiroptera).

Science.gov (United States)

Cabral, A D; Gama, A R; Sodré, M M; Savani, E S M M; Galvão-Dias, M A; Jordão, L R; Maeda, M M; Yai, L E O; Gennari, S M; Pena, H F J

2013-03-31

There are currently no reports on the isolation and molecular examination of Toxoplasma gondii from bats. Here, we report the isolation and genotypic characterisation of two T. gondii isolates from bats. A total of 369 bats from different municipalities in São Paulo state, southeastern Brazil, were captured and euthanised, and collected tissues (heart and pectoral muscle) were processed for each bat or in pools of two or three bats and bioassayed in mice (a total of 283 bioassays). Eleven PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) markers were used to genotype positive samples: SAG1, SAG2 (5'-3'SAG2 and alt. SAG2), SAG3, BTUB, GRA6, L358, c22-8, c29-2, PK1, CS3 and Apico. The parasite was isolated from two bats from São Paulo city: an insectivorous bat, the velvety free-tailed bat Molossus molossus, and a hematophagous bat, the common vampire bat Desmodus rotundus. Isolates were designated TgBatBr1 and TgBatBr2, respectively. The genotype of the isolate from M. molossus (TgBatBr1) has been previously described in an isolate from a capybara from São Paulo state, and the genotype from the D. rotundus isolate (TgBatBr2) has already been identified in isolates from cats, chickens, capybaras, sheep, a rodent and a common rabbit from different Brazilian states, suggesting that this may be a common T. gondii lineage circulating in some Brazilian regions. Isolation of T. gondii from a hematophagous species is striking. This study reveals that bats can share the same isolates that are found in domesticated and wild terrestrial animals. This is the first report of the isolation and genotyping of T. gondii in chiropterans. Copyright © 2012 Elsevier B.V. All rights reserved.

Toxoplasma gondii exposure and Parkinson's disease: a case–control study

Science.gov (United States)

Alvarado-Esquivel, Cosme; Méndez-Hernández, Edna Madai; Salas-Pacheco, José Manuel; Ruano-Calderón, Luis Ángel; Hernández-Tinoco, Jesús; Arias-Carrión, Oscar; Sánchez-Anguiano, Luis Francisco; Castellanos-Juárez, Francisco Xavier; Sandoval-Carrillo, Ada Agustina; Liesenfeld, Oliver; Ramos-Nevárez, Agar

2017-01-01

Objectives To determine the association between Toxoplasma gondii infection and Parkinson's disease and to investigate whether T. gondii seropositivity is associated with the general characteristics of patients with Parkinson's disease. Design Case–control study. Setting Cases and controls were enrolled in Durango City, Mexico. Participants 65 patients with Parkinson's disease and 195 age- and gender-matched control subjects without Parkinson's disease. Primary and secondary outcome measures Serum samples of participants were analysed for anti-T. gondii IgG and IgM antibodies by commercially available enzyme-linked immunoassays. Prevalence of T. gondii DNA was determined in seropositive subjects using PCR. The association between clinical data and infection was examined by bivariate analysis. Results Anti-T. gondii IgG antibodies were found in 6/65 cases (9.2%) and in 21/195 controls (10.8%) (OR 0.84; 95% CI 0.32 to 2.18; p=0.81). The frequency of high (>150 IU/mL) antibody levels was similar among cases and controls (p=0.34). None of the anti-T. gondii IgG positive cases and four of the anti-T. gondii IgG positive controls had anti-T. gondii IgM antibodies (p=0.54). The prevalence of T. gondii DNA was comparable in seropositive cases and controls (16.7% and 25%, respectively; p=1.0). Seroprevalence of T. gondii infection was associated with a young age onset of disease (p=0.03), high Unified Parkinson Disease Rating Scale scores (p=0.04) and depression (p=0.02). Seropositivity to T. gondii infection was lower in patients treated with pramipexole than in patients without this treatment (p=0.01). However, none of the associations remained significant after Bonferroni correction. Conclusions The results do not support an association between T. gondii infection and Parkinson's disease. However, T. gondii infection might have an influence on certain symptoms of Parkinson's disease. Further research to elucidate the role of T. gondii exposure on Parkinson's disease

Antibody detection and molecular characterization of toxoplasma gondii from bobcats (Lynx rufus), domestic cats (Felis catus), and wildlife from Minnesota, USA

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Little is known of the epidemiology of toxoplasmosis in Minnesota. In this study, we evaluated Toxoplasma gondii infection in 50 wild bobcats (Lynx rufus) and 75 other animals on/near 10 cattle farms. Antibodies to T. gondii were assayed in serum samples or tissue fluids by the modified agglutinatio...

Dual Identification and Analysis of Differentially Expressed Transcripts of Porcine PK-15 Cells and Toxoplasma gondii during in vitro Infection

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Chun-Xue eZhou

2016-05-01

Full Text Available Toxoplasma gondii is responsible for causing toxoplasmosis, one of the most prevalent zoonotic parasitoses worldwide. The mechanisms that mediate T. gondii infection of pigs (the most common source of human infection and renal tissues are still unknown. To identify the critical alterations that take place in the transcriptome of both porcine kidney (PK-15 cells and T. gondii following infection, infected cell samples were collected at 1, 3, 6, 9, 12, 18, and 24 h post infection and RNA-Seq data were acquired using Illumina Deep Sequencing. Differential Expression of Genes (DEGs analysis was performed to study the concomitant gene-specific temporal patterns of induction of mRNA expression of PK-15 cells and T. gondii. High sequence coverage enabled us to thoroughly characterize T. gondii transcriptome and identify the activated molecular pathways in host cells. More than 6G clean bases/ sample, including > 40 million clean reads were obtained. These were aligned to the reference genome of T. gondii and wild boar (Sus scrofa. DEGs involved in metabolic activities of T. gondii showed time-dependent down-regulation. However, DEGs involved in immune or disease related pathways of PK-15 cells peaked at 6 h PI, and were highly enriched as evidenced by KEGG analysis. Protein-protein interaction analysis revealed that TGME49_120110 (PCNA, TGME49_049180 (DHFR-TS, TGME49_055320 and TGME49_002300 (ITPase are the four hub genes with most interactions with T. gondii at the onset of infection. These results reveal altered profiles of gene expressed by PK-15 cells and T. gondii during infection and provide the groundwork for future virulence studies to uncover the mechanisms of T. gondii interaction with porcine renal tissue by functional analysis of these DEGs.

Construction and characterization of HIV type 1 CRF07_BC infectious molecular clone from men who have sex with men.

Science.gov (United States)

Jiang, Yan-Ling; Bai, Wen-Wei; Qu, Fan-Wei; Ma, Hua; Jiang, Run-Sheng; Shen, Bao-Sheng

2016-03-01

This study aimed to investigate the biological characterization of HIV type 1 (HIV-1) CRF07_BC infection among men who have sex with men (MSM). From November 2011 to November 2013, a total of 66 blood samples were collected from MSM with acute HIV-1 infection with CRF07_BC subgroup strains. Deletion in the gag p6 region was detected by sequence alignment and comparative analysis. Peripheral blood mononuclear cells (PBMCs) of HNXX1301-1307 samples were separated by density gradient centrifugation. Nested polymerase chain reaction (nPCR) was used to amplify the viral DNA. The near full-length HIV-1 DNA products were ligated to the long terminal repeat (LTR) vector plasmid (07BCLTR) to construct a full-length HIV clone. The molecular clone was transfected into HEK-293T cells, TZM-b1 cells and patients' PBMCs. The pregenome of an infectious molecular clone of HIV-1 (pNL4-3) was amplified, and a subclone with CRF07_BC was developed to construct the full-length chimeric molecular clone pNL4-3/07BCLTR. Detection of p24 antigen and luciferase activity was used to measure the in vitro infectivity of pNL4-3/07BCLTR. Among the 66 MSM patients infected with CRF07_BC strains, deletion mutations of the Gag P6 proteins were found in 7 of 18CRF07_BC strains; deletion mutations of 2-13 amino acids in different regions were discovered in 6 strains; and the remaining 42 strains did not show deletions. Seven strains with amino acids deficiency in the P6 protein accounted for 27% of all strains and 75% of all deletion genotype strains. A total of 186 full-length molecular clones of CRF07_BC were constructed. There were 5, 9, 10 and 11 clones of HNXX1302, HNXX1304, HNXX1305 and HNXX1306 that resulted in p24-positive supernatant when transfected into HEK-293T cells. Full-length clones of HNXX1302, HNXX1304, HNXX1305 and HNXX1306 showed slight infection in the transfected TZM-b1 cells, as judged by the fluorescence values of TZM-b1 cells 48h post-transfection. However, we were unable to

Occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chicken breeding in Northeast, Brazil

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Marcela Fernanda Torres Samico Fernandes

2016-03-01

Full Text Available Abstract The aim of the present study was to investigate the occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chickens bred in the metropolitan area of Recife, Brazil. In total, 212 serum samples were collected from 16 properties, and 12 backyard chickens were collected in the six sanitary districts of Recife. An indirect immunofluorescence assay (IFA was used to investigate the occurrence of anti-Toxoplasma gondii antibodies. Polymerase chain reaction (PCR was used to detect T. gondii DNA in brain, heart, liver and lung specimens. Of the samples analyzed by serology, 86/212 (40.56% were positive; of the samples analyzed by PCR, 2/12 (16.7% were positive, with both samples positive by both tests (serological and molecular. The presence of antibody anti-T. gondii and parasite DNA in tissues of these animals are worrying aspects for public health because there is a risk of transmission of the parasite to humans through eating undercooked or raw meat. Based on the results, the adoption of preventive measures to prevent the cats access to the chickens creations should be encouraged, since these animals were identified in most of the studied properties.

Dissecting the molecular assembly of the Toxoplasma gondii MyoA motility complex.

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Powell, Cameron J; Jenkins, Meredith L; Parker, Michelle L; Ramaswamy, Raghavendran; Kelsen, Anne; Warshaw, David M; Ward, Gary E; Burke, John E; Boulanger, Martin J

2017-11-24

Apicomplexan parasites such as Toxoplasma gondii rely on a unique form of locomotion known as gliding motility. Generating the mechanical forces to support motility are divergent class XIV myosins (MyoA) coordinated by accessory proteins known as light chains. Although the importance of the MyoA-light chain complex is well-established, the detailed mechanisms governing its assembly and regulation are relatively unknown. To establish a molecular blueprint of this dynamic complex, we first mapped the adjacent binding sites of light chains MLC1 and ELC1 on the MyoA neck (residues 775-818) using a combination of hydrogen-deuterium exchange mass spectrometry and isothermal titration calorimetry. We then determined the 1.85 Å resolution crystal structure of MLC1 in complex with its cognate MyoA peptide. Structural analysis revealed a bilobed architecture with MLC1 clamping tightly around the helical MyoA peptide, consistent with the stable 10 nm K d measured by isothermal titration calorimetry. We next showed that coordination of calcium by an EF-hand in ELC1 and prebinding of MLC1 to the MyoA neck enhanced the affinity of ELC1 for the MyoA neck 7- and 8-fold, respectively. When combined, these factors enhanced ELC1 binding 49-fold (to a K d of 12 nm). Using the full-length MyoA motor (residues 1-831), we then showed that, in addition to coordinating the neck region, ELC1 appears to engage the MyoA converter subdomain, which couples the motor domain to the neck. These data support an assembly model where staged binding events cooperate to yield high-affinity complexes that are able to maximize force transduction. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Characterization of nonprimate hepacivirus and construction of a functional molecular clone

DEFF Research Database (Denmark)

Scheel, Troels K H; Kapoor, Amit; Nishiuchi, Eiko

2015-01-01

Nonprimate hepacivirus (NPHV) is the closest known relative of hepatitis C virus (HCV) and its study could enrich our understanding of HCV evolution, immunity, and pathogenesis. High seropositivity is found in horses worldwide with ∼ 3% viremic. NPHV natural history and molecular virology remain...... circulating liver enzymes and mild hepatitis was observed, followed by viral clearance. This establishes the molecular components of a functional NPHV genome. Thus, NPHV appears to resemble HCV not only in genome structure but also in its ability to establish chronic infection with delayed seroconversion...... and hepatitis. This NPHV infectious clone and resulting acute phase sera will facilitate more detailed studies on the natural history, pathogenesis, and immunity of this novel hepacivirus in its natural host....

Molecular detection of Toxoplasma gondii in water samples from Scotland and a comparison between the 529bp real-time PCR and ITS1 nested PCR.

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Wells, Beth; Shaw, Hannah; Innocent, Giles; Guido, Stefano; Hotchkiss, Emily; Parigi, Maria; Opsteegh, Marieke; Green, James; Gillespie, Simon; Innes, Elisabeth A; Katzer, Frank

2015-12-15

Waterborne transmission of Toxoplasma gondii is a potential public health risk and there are currently no agreed optimised methods for the recovery, processing and detection of T. gondii oocysts in water samples. In this study modified methods of T. gondii oocyst recovery and DNA extraction were applied to 1427 samples collected from 147 public water supplies throughout Scotland. T. gondii DNA was detected, using real time PCR (qPCR) targeting the 529bp repeat element, in 8.79% of interpretable samples (124 out of 1411 samples). The samples which were positive for T. gondii DNA originated from a third of the sampled water sources. The samples which were positive by qPCR and some of the negative samples were reanalysed using ITS1 nested PCR (nPCR) and results compared. The 529bp qPCR was the more sensitive technique and a full analysis of assay performance, by Bayesian analysis using a Markov Chain Monte Carlo method, was completed which demonstrated the efficacy of this method for the detection of T. gondii in water samples. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Crioconservación de toxoplasma gondii

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Sofía Duque Beltrán

1992-03-01

Full Text Available Toxoplasma gondii fue crioconservado en nitrógeno líquido usando como preservativo glicerol al 10% con el fin de mantener el protozoo por un largo período de tiempo. El descongelamiento de T. gondii se llevó a cabo, cuando los parásitos fueron requeridos para uso como antígeno, a los 10, 40 y 270 días siguientes a su crioconservación. La viabilidad y patogenicidad del parásito fue confirmada in vivo. La crioconservación de T. gondii disminuyó los costos de mantenimiento in vivo y de recursos humanos tanto en el bioterio como en el laboratorio.

Molecular cloning and characterization of Izumo1 gene from bovine testis

OpenAIRE

Kim, Ekyune

2015-01-01

A well-characterized sperm specific protein of the Member of immunoglobulin superfamily, IZUMO1, has crucial role in fertilization by mediating sperm binding to the egg plasma membrane in the mouse. However little is known about IZUMO1 in bovine. Here, we describe the molecular cloning and expression analysis of bovine IZUMO1 (bIZUMO1). RT-PCR and Western blot analysis of the bovine tissues indicated that bIZUMO1 was specifically expressed in the testis and sperm, Furthermore, the result of o...

Toxoplasma gondii infection in humans in China

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He Shenyi

2011-08-01

Full Text Available Abstract Toxoplasmosis is a zoonotic infection of humans and animals, caused by the opportunistic protozoan Toxoplasma gondii, a parasite belonging to the phylum Apicomplexa. Infection in pregnant women may lead to abortion, stillbirth or other serious consequences in newborns. Infection in immunocompromised patients can be fatal if not treated. On average, one third of people are chronically infected worldwide. Although very limited information from China has been published in the English journals, T. gondii infection is actually a significant human health problem in China. In the present article, we reviewed the clinical features, transmission, prevalence of T. gondii infection in humans in China, and summarized genetic characterizations of reported T. gondii isolates. Educating the public about the risks associated with unhealthy food and life style habits, tracking serological examinations to special populations, and measures to strengthen food and occupational safety are discussed.

Loop-mediated isothermal amplification (LAMP): early detection of Toxoplasma gondii infection in mice.

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Kong, Qing-Ming; Lu, Shao-Hong; Tong, Qun-Bo; Lou, Di; Chen, Rui; Zheng, Bin; Kumagai, Takashi; Wen, Li-Yong; Ohta, Nobuo; Zhou, Xiao-Nong

2012-01-03

Toxoplasmosis is a widespread zoonotic parasitic disease that occurs in both animals and humans. Traditional molecular assays are often difficult to perform, especially for the early diagnosis of Toxoplasma gondii infections. Here, we established a novel loop-mediated isothermal amplification targeting the 529 bp repeat element (529 bp-LAMP) to detect T. gondii DNA in blood samples of experimental mice infected with tachyzoites of the RH strain. The assay was performed with Bst DNA polymerase at 65°C for 1 h. The detection limit of the 529 bp-LAMP assay was as low as 0.6 fg of T. gondii DNA. The sensitivity of this assay was 100 and 1000 fold higher than that of the LAMP targeting B1 gene (B1-LAMP) and nested PCR targeting 529 bp repeat element (529 bp-nested PCR), respectively. The specificity of the 529 bp-LAMP assay was determined using the DNA samples of Trypanosoma evansi, Plasmodium falciparum, Paragonimus westermani, Schistosoma japonicum, Fasciola hepatica and Angiostrongylus cantonensis. No cross-reactivity with the DNA of any parasites was found. The assay was able to detect T. gondii DNA in all mouse blood samples at one day post infection (dpi). We report the following findings: (i) The detection limit of the 529 bp-LAMP assay is 0.6 fg of T. gondii DNA; (ii) The assay does not involve any cross-reactivity with the DNA of other parasites; (iii) This is the first report on the application of the LAMP assay for early diagnosis of toxoplasmosis in blood samples from experimentally infected mice. Due to its simplicity, sensitivity and cost-effectiveness for common use, we suggest that this assay should be used as an early diagnostic tool for health control of toxoplasmosis.

Bovine viral diarrhea virus: molecular cloning of genomic RNA and its diagnostic application

International Nuclear Information System (INIS)

Brock, K.V.

1987-01-01

Molecular cloning of a field isolate of bovine viral diarrhea virus (BVDV) strain 72 RNA was done in this study. The sensitivity and specificity of cloned cDNA sequences in hybridization assays with various BVDV strains were determined. cDNA was synthesized from polyadenylated BVDV RNA templates with oligo-dT primers, reverse transcriptase, and DNA polymerase I. The newly synthesized double-stranded BVDV cDNA was C-tailed with terminal deoxytransferase and annealed into G-tailed, Pst-1-cut pUC9 plasmid. Escherichia coli was transformed with the recombinant plasmids and a library of approximately 200 BVDV specific cDNA clones varying in length from 0.5 to 2.6 kilobases were isolated. The sensitivity and specificity of hybridization between the labelled cDNA and BVDV target sequences were determined. Cloned BVDV sequences were isolated from pUC9 plasmid DNA and labelled with 32 P by nick translation. The detection limit by dot blot hybridization assay was 20 pg of purified genomic BVDV RNA. cDNA hybridization probes were specific for all strains of BVDV tested, regardless of whether they were noncytopathic and cytopathic, but did not hybridize with heterologous bovine viruses tested. Probes did not hybridize with uninfected cell culture or cellular RNA. Hybridization probes were at least as sensitive as infectivity assays in detecting homologous virus

Genetic diversity of Toxoplama gondii isolates from Ethiopian feral cats

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Recent studies indicate greater genetic variability among isolates of Toxoplasma gondii worldwide than previously thought. However, there is no information on genetic diversity of T. gondii from any host in Ethiopia. In the present study, genotyping was performed on viable T. gondii isolates by bioa...

Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for detection of Toxoplasma gondii in the environment.

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Wu, Y D; Xu, M J; Wang, Q Q; Zhou, C X; Wang, M; Zhu, X Q; Zhou, D H

2017-08-30

Toxoplasma gondii infects all warm-blooded vertebrates, resulting in a great threat to human health and significant economic loss to the livestock industry. Ingestion of infectious oocysts of T. gondii from the environment is the major source of transmission. Detection of T. gondii oocysts by existing methods is laborious, time-consuming and expensive. The objective of the present study was to develop a recombinase polymerase amplification (RPA) method combined with a lateral flow (LF) strip for detection of T. gondii oocysts in the soil and water. The DNA of T. gondii oocysts was amplified by a pair of specific primers based on the T. gondii B1 gene over 15min at a constant temperature ranging from 30°C to 45°C using RPA. The amplification product was visualized by the lateral flow (LF) strip within 5min using the specific probe added to the RPA reaction system. The sensitivity of the established assay was 10 times higher than that of nested PCR with a lower detection limit of 0.1 oocyst per reaction, and there was no cross-reactivity with other closely related protozoan species. Fifty environmental samples were further assessed for the detection validity of the LF-RPA assay (B1-LF-RPA) and compared with nested PCR based on the B1 gene sequence. The B1-LF-RPA and nested PCR both showed that 5 out of the 50 environmental samples were positive. The B1-LF-RPA method was also proven to be sufficiently tolerant of existing inhibitors in the environment. In addition, the advantages of simple operation, speediness and cost-effectiveness make B1-LF-RPA a promising molecular detection tool for T. gondii. Copyright © 2017 Elsevier B.V. All rights reserved.

Isolation and Genotyping of Toxoplasma gondii in Brazilian Dogs.

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da Silva, Jamille Rodrigues; Maciel, Bianca Mendes; de Santana Souza Santos, Luana Karla Nogueira; Carvalho, Fábio Santos; de Santana Rocha, Daniele; Lopes, Carlos Wilson Gomes; Albuquerque, George Rêgo

2017-06-01

Strains of Toxoplasma gondii in Brazil are highly genetically diverse compared to strains from North America and Europe. Dogs are epidemiologically important because they act as sentinels for T. gondii infections in humans and are good indicators of environmental contamination. The aim of this study was to isolate and genetically characterize T. gondii strains from tissues of naturally infected Brazilian dogs. For this study, 21 blood samples were collected from dogs at the Zoonosis Control Centers of Ilhéus and Itabuna cities, Bahia, Brazil. The sera were examined for T. gondii antibodies using the indirect hemagglutination test. Brains and hearts of seropositive dogs were bioassayed in mice to isolate and characterize T. gondii parasites by PCR-RFLP using 10 genetic markers (SAG1, newSAG2, SAG3, BTUB, c22-8, c29-2, GRA6, PK1, APICO, and L358). However, T. gondii was isolated from only 4 (57.1%) dogs, designated TgDgBr6, 13, 17, and 21. All strains were virulent, causing clinical changes (rough hair coat, lethargy, and abdominal distention) and the death of all mice within 8-20 days after inoculation. Genetic analysis of these 4 T. gondii isolates revealed 4 distinct genotypes with different clonal lineage combinations (types I, II, and III) and 2 atypical alleles. Using PCR-RFLP with several markers, this study contributes to evaluations of the genetic diversity of strains circulating in Brazil.

Sexual transmission of Toxoplasma gondii in sheep.

Science.gov (United States)

Lopes, Welber Daniel Zanetti; Rodriguez, Joana D'Ark; Souza, Fernando A; dos Santos, Thais Rabelo; dos Santos, Ricardo Silva; Rosanese, Walter Matheus; Lopes, Werik Renato Zanetti; Sakamoto, Cláudio Alessandro; da Costa, Alvimar José

2013-07-01

Male sheep of reproductive age were distributed into three groups: GI, a sheep inoculated (oral) with 2.0×10(5) oocysts of the P strain of Toxoplasma gondii; GII, a sheep infected (subcutaneous) with 1.0×10(6) tachyzoites of the RH strain of T. gondii; and GIII, a sheep kept as a control (not infected). After the inoculation of the males, 12 breeding ewes, which were not pregnant and which were serologically negative for reproductive diseases (particularly toxoplasmosis), were distributed into three groups, synchronized, and subsequently exposed to natural mating with previously inoculated males. The distribution was as follows: five ewes that underwent natural mating with the GI male, five ewes that were exposed to natural mating with the GII male, and two ewes that were mated with the non-infected male (control). Serum samples of all the ewes were collected on days -30, -14, -7, -1, and 0 (days before natural mating) and on days 1, 3, 5, 7, 11, 14, and weekly until birth; the presence of serum antibodies against T. gondii was assessed by IFAT. Using a bioassay and PCR, T. gondii was isolated from the semen of the infected reproducing sheep before mating. Following natural mating, 5 of the 12 females displayed antibodies specific for T. gondii; of these animals, two of the ewes underwent natural mating with the male inoculated with oocysts (GI) and three with the male infected with tachyzoites (GII). One of the females that displayed antibodies specific to this coccidian and that underwent natural mating with the GII sheep had a macerated fetus on the 70th day following coverage. Using a bioassay after the birth, it was possible to isolate T. gondii from samples of the "pool" of tissues from the five females that seroconverted after natural mating and from their respective lambs. Using PCR, the DNA of T. gondii was isolated from the "pool" of tissues from one and two females exposed to natural mating with the reproductive males infected with the oocysts and

Seroprevalence of Toxoplasma gondii infection amongst residents ...

African Journals Online (AJOL)

Toxoplasmosis is a zoonotic disease, recognized as a serious public health problem worldwide. Toxoplasma gondii infection has become a major public health concern in recent years due to the ravaging HIV/AIDS pandemic. A serological survey was carried out in Tanga district of north-eastern Tanzania to assess T. gondii ...

Global iTRAQ-based proteomic profiling of Toxoplasma gondii oocysts during sporulation.

Science.gov (United States)

Zhou, Chun-Xue; Zhu, Xing-Quan; Elsheikha, Hany M; He, Shuai; Li, Qian; Zhou, Dong-Hui; Suo, Xun

2016-10-04

Toxoplasma gondii is a medically and economically important protozoan parasite. However, the molecular mechanisms of its sporulation remain largely unknown. Here, we applied iTRAQ coupled with 2D LC-MS/MS proteomic analysis to investigate the proteomic expression profile of T. gondii oocysts during sporulation. Of the 2095 non-redundant proteins identified, 587 were identified as differentially expressed proteins (DEPs). Based on Gene Ontology enrichment and KEGG pathway analyses the majority of these DEPs were found related to the metabolism of amino acids, carbon and energy. Protein interaction network analysis generated by STRING identified ATP-citrate lyase (ACL), GMP synthase, IMP dehydrogenase (IMPDH), poly (ADP-ribose) glycohydrolase (PARG), and bifunctional dihydrofolate reductase-thymidylate synthase (DHFR-TS) as the top five hubs. We also identified 25 parasite virulence factors that were expressed at relatively high levels in sporulated oocysts compared to non-sporulated oocysts, which might contribute to the infectivity of mature oocysts. Considering the importance of oocysts in the dissemination of toxoplasmosis these findings may help in the search of protein targets with a key role in infectiousness and ecological success of oocysts, creating new opportunities for the development of better means for disease prevention. The development of new preventative interventions against T. gondii infection relies on an improved understanding of the proteome and chemical pathways of this parasite. To identify proteins required for the development of environmentally resistant and infective T. gondii oocysts, we compared the proteome of non-sporulated (immature) oocysts with the proteome of sporulated (mature, infective) oocysts. iTRAQ 2D-LC-MS/MS analysis revealed proteomic changes that distinguish non-sporulated from sporulated oocysts. Many of the differentially expressed proteins were involved in metabolic pathways and 25 virulence factors were identified

Toxoplasma gondii coinfection with diseases and parasites in wild rabbits in Scotland.

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Mason, Sam; Dubey, J P; Smith, Judith E; Boag, Brian

2015-09-01

In wild rabbits (Oryctolagus cuniculus) on an estate in Perthshire, central Scotland, the seroprevalence of Toxoplasma gondii was 18/548 (3·3%). The wild rabbit could be a T. gondii reservoir and it has potential value as a sentinel of T. gondii in environmental substrates. Toxoplasma gondii was associated with female sex (P myxomatosis caused by the virus Myxomatosis cuniculi, the intensity of roundworm eggs, the year or season, rabbit age or distance from farm buildings. Coinfections could have been affected by gestational down regulation of type 1 T helper cells. A sudden influx or release of T. gondii oocysts might have occurred. This is the first report of T. gondii in any wild herbivore in Scotland and also the first report of lapine T. gondii as a coinfection with E. stiedae, M. cuniculi and helminths.

Toxoplasma gondii sequesters lysosomes from mammalian hosts in the vacuolar space.

Science.gov (United States)

Coppens, Isabelle; Dunn, Joe Dan; Romano, Julia D; Pypaert, Marc; Zhang, Hui; Boothroyd, John C; Joiner, Keith A

2006-04-21

The intracellular compartment harboring Toxoplasma gondii satisfies the parasite's nutritional needs for rapid growth in mammalian cells. We demonstrate that the parasitophorous vacuole (PV) of T. gondii accumulates material coming from the host mammalian cell via the exploitation of the host endo-lysosomal system. The parasite actively recruits host microtubules, resulting in selective attraction of endo-lysosomes to the PV. Microtubule-based invaginations of the PV membrane serve as conduits for the delivery of host endo-lysosomes within the PV. These tubular conduits are decorated by a parasite coat, including the tubulogenic protein GRA7, which acts like a garrote that sequesters host endocytic organelles in the vacuolar space. These data define an unanticipated process allowing the parasite intimate and concentrated access to a diverse range of low molecular weight components produced by the endo-lysosomal system. More generally, they identify a unique mechanism for unidirectional transport and sequestration of host organelles.

Adenosine metabolism in Toxoplasma gondii: potential targets for chemotherapy.

Science.gov (United States)

el Kouni, Mahmoud H

2007-01-01

Toxoplasma gondii is an intracellular parasitic protozoan that infects approximately a billion people worldwide. Infection with T. gondii represents a major health problem for immunocompromised individuals, such as AIDS patients, organ transplant recipients, and the unborn children of infected mothers. Currently available drugs usually do not eradicate infection and as many as 50% of the patients do not respond to this therapy. Furthermore, they are ineffective against T. gondii tissue cysts. In addition, prolonged exposure to these drugs induces serious host toxicity forcing the discontinuation of the therapy. Finally, there is no effective vaccine currently available for the treatment of toxoplasmosis. Therefore, it is necessary to develop new and effective drugs for the treatment and management of toxoplasmosis. The rational design of a drug depends on the exploitation of fundamental biochemical or physiological differences between pathogens and their host. Some of the most striking differences between T. gondii and their mammalian host are found in purine metabolism. T. gondii, like most parasites studied, lack the ability to synthesize purines do novo and depend on the salvage of purines from their host to satisfy their requirements of purines. In this respect, the salvage of adenosine is the major source of purines in T. gondii. Therefore, interference with adenosine uptake and metabolism in T. gondii can be selectively detrimental to the parasite. The host cells, on the other hand, can still obtain their purine requirements by their de novo pathways. This review will focus on the broad aspects of the adenosine transport and the enzyme adenosine kinase (EC 2.7.1.20) which are the two primary routes for adenosine utilization in T. gondii, in an attempt to illustrate their potentials as targets for chemotherapy against this parasite.

Development of RAPD-SCAR markers for different Ganoderma species authentication by improved RAPD amplification and molecular cloning.

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Fu, J J; Mei, Z Q; Tania, M; Yang, L Q; Cheng, J L; Khan, M A

2015-05-25

The sequence-characterized amplified region (SCAR) is a valuable molecular technique for the genetic identification of any species. This method is mainly derived from the molecular cloning of the amplified DNA fragments achieved from the random amplified polymorphic DNA (RAPD). In this study, we collected DNA from 10 species of Ganoderma mushroom and amplified the DNA using an improved RAPD technique. The amplified fragments were then cloned into a T-vector, and positive clones were screened, indentified, and sequenced for the development of SCAR markers. After designing PCR primers and optimizing PCR conditions, 4 SCAR markers, named LZ1-4, LZ2-2, LZ8-2, and LZ9-15, were developed, which were specific to Ganoderma gibbosum (LZ1-4 and LZ8-2), Ganoderma sinense (LZ2-2 and LZ8-2), Ganoderma tropicum (LZ8-2), and Ganoderma lucidum HG (LZ9-15). These 4 novel SCAR markers were deposited into GenBank with the accession Nos. KM391935, KM391936, KM391937, and KM391938, respectively. Thus, in this study we developed specific SCAR markers for the identification and authentication of different Ganoderma species.

Toxoplasma gondii in wild and domestic animals from New Caledonia

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Roqueplo C.

2011-11-01

Full Text Available Samples (serum or meat juice collected from 205 animals in New Caledonia in April 2009 were tested for antibodies against Toxoplasma gondii by ELISA using the multi-species ID Screen® Toxoplasmosis Indirect kit (IDVET, Montpellier. Antibodies to T. gondii were detected in 2% (1/49 of the pigs, in 3.3% (1/30 of the cattle, in 13.8% (4/29 of Rusa deers, in 16% (4/25 of the horses, in 32.8% (21/64 of the dogs, and in 50% (4/8 of cats. Statistically, no significant difference was observed between T. gondii seroprevalence and age or sex. No survey on the prevalence of T. gondii in animals has ever been conducted in New Caledonia and this is the first serological evidence of T. gondii in Rusa deer (Cervus timorensis russa. These results indicate an important circulation of T. gondii exists in the animal populations of New Caledonia. In view of humans being exposed, it is advisable to insist on sanitary education and on respect for good hygienic and food practice.

Molecular cloning and expression of Corynebacterium glutamicum genes for amino acid synthesis in Escherichia coli cells

International Nuclear Information System (INIS)

Beskrovnaya, O.Yu.; Fonshtein, M.Yu.; Kolibaba, L.G.; Yankovskii, N.K.; Debabov, V.G.

1989-01-01

Molecular cloning of Corynebacterium glutamicum genes for threonine and lysine synthesis has been done in Escherichia coli cells. The clonal library of EcoRI fragments of chromosomal DNA of C. glutamicum was constructed on the plasmid vector λpSL5. The genes for threonine and lysine synthesis were identified by complementation of E. coli mutations in thrB and lysA genes, respectively. Recombinant plasmids, isolated from independent ThrB + clone have a common 4.1-kb long EcoRI DNA fragment. Hybrid plasmids isolated from LysA + transductants of E. coli have common 2.2 and 3.3 kb long EcoRI fragments of C. glutamicum DNA. The hybrid plasmids consistently transduced the markers thrB + and lysA + . The Southern hybridization analysis showed that the cloned DNA fragments hybridized with the fragments of identical length in C. glutamicum chromosomes

Evasion of Human Neutrophil-Mediated Host Defense during Toxoplasma gondii Infection.

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Lima, Tatiane S; Gov, Lanny; Lodoen, Melissa B

2018-02-13

Neutrophils are a major player in host immunity to infection; however, the mechanisms by which human neutrophils respond to the intracellular protozoan parasite Toxoplasma gondii are still poorly understood. In the current study, we found that, whereas primary human monocytes produced interleukin-1beta (IL-1β) in response to T. gondii infection, human neutrophils from the same blood donors did not. Moreover, T. gondii inhibited lipopolysaccharide (LPS)-induced IL-1β synthesis in human peripheral blood neutrophils. IL-1β suppression required active parasite invasion, since heat-killed or mycalolide B-treated parasites did not inhibit IL-1β release. By investigating the mechanisms involved in this process, we found that T. gondii infection of neutrophils treated with LPS resulted in reduced transcript levels of IL-1β and NLRP3 and reduced protein levels of pro-IL-1β, mature IL-1β, and the inflammasome sensor NLRP3. In T. gondii -infected neutrophils stimulated with LPS, the levels of MyD88, TRAF6, IKKα, IKKβ, and phosphorylated IKKα/β were not affected. However, LPS-induced IκBα degradation and p65 phosphorylation were reduced in T. gondii- infected neutrophils, and degradation of IκBα was reversed by treatment with the proteasome inhibitor MG-132. Finally, we observed that T. gondii inhibited the cleavage and activity of caspase-1 in human neutrophils. These results indicate that T. gondii suppression of IL-1β involves a two-pronged strategy whereby T. gondii inhibits both NF-κB signaling and activation of the NLRP3 inflammasome. These findings represent a novel mechanism of T. gondii evasion of human neutrophil-mediated host defense by targeting the production of IL-1β. IMPORTANCE Toxoplasma gondii is an obligate intracellular parasite that infects approximately one-third of humans worldwide and can invade virtually any nucleated cell in the human body. Although it is well documented that neutrophils infiltrate the site of acute T. gondii

Toxoplasma gondii gamma irradiation using Co-60

International Nuclear Information System (INIS)

Serra-Freire, Nicolau Maues

1996-01-01

The use of nuclear power through radiation for the destruction of microorganisms which cause food deterioration, infections and toxicosis, is specifically for peaceful purposes. Toxoplasma gondii is a protozoa responsible for illnesses in humans and animals. One of the most common ways of transmission is through raw or poorly cooked meat. There is little information on the resistance of T. gondii to radiation. The objective of this research is to determine the Minimum Lethal Dose (MLD) of gamma radiation for those microorganisms. Suspensions of T. gondii containing approximately one million taquizoites/ml were irradiated with doses between up 0,01 up to 0,15 kGy (Kilogray) and inoculated to mice. The surviving T. gondii were re-irradiated with 0,01 up to 0,16 kGy. The irradiated protozoa were totally destroyed with a 0,15 kGy dose (MLD). Taquizoites issued from live protozoa of 0,14 kGy also were completely destroyed with dose of 0,15 kGy. No increase in resistance was observed regarding the non irradiated protozoa. (author)

First report of Toxoplasma gondii seroprevalence in pet parrots in China.

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Zhang, Xiao-Xuan; Zhang, Nian-Zhang; Tian, Wei-Peng; Zhou, Dong-Hui; Xu, Ying-Tian; Zhu, Xing-Quan

2014-06-01

Toxoplasmosis, caused by the obligate intracellular protozoan parasite Toxoplasma gondii, has become a serious public health problem worldwide. T. gondii can infect almost all warm-blooded animals, including parrots. However, little is known of T. gondii infection in parrots in China. Antibodies against T. gondii in 311 parrots including 202 Budgerigars (Melopsittacus undulatus), 26 Lovebirds (Agapornis sp.), 22 Cockatiels (Nymphicus hollandicus), and 61 Alexandrine Parakeets (Psittacula eupatria) in the cities of Beijing and Weifang in north China were tested using the modified agglutination test (MAT). Twenty-six (8.36%) out of 311 serum samples were positive for T. gondii at the cutoff of 1:5. Among the four species, a higher seroprevalence of T. gondii was found in Cockatiels (13.64%, 95% confidence interval [CI] 0.00-27.98), although the difference was not statistically significant (p=0.61). Seropositivity rates against T. gondii in male parrots (10.43%, 95% CI 5.74-15.12) were not statistically different from that in female parrots (6.08%, 95% CI 2.23-9.93, p=0.17). The seropositivity of T. gondii in parrots from Weifang and Beijing was 11.11% (95% CI 6.13-16.09) and 5.70% (95% CI 2.08-9.31), respectively. The seroprevalence varied in parrots of different age groups, ranging from 5.71% (95% CI 1.27-10.15) to 13.00% (95% CI 6.41-19.69), however, the difference among age groups was not statistically significant (p=0.12). The seroprevalence of T. gondii infection in parrots in summer (11.63%, 95% CI 6.84-16.42) was significantly higher than in spring (4.32%, 95% CI 0.94-7.70, p=0.02). The results of the present survey indicated that parrots in China are exposed to T. gondii. To our knowledge, this is the first report of T. gondii seroprevalence in parrots in China.

Seroprevalence of Toxoplasma gondii and Neospora caninum in ...

African Journals Online (AJOL)

Background: Toxoplasma gondii and Neospora caninum are protozoans infecting a wide range of mammals; the etiologic agents of Toxoplasmosis and Neosporosis respectively, This study investigated the prevalence of antibodies to Toxoplasma gondii and Neospora caninum in dogs from southwestern Nigeria. Materials ...

Genetic diversity of Toxoplasma gondii isolates from Ethiopian feral cats.

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Dubey, J P; Choudhary, S; Tilahun, G; Tiao, N; Gebreyes, W A; Zou, X; Su, C

2013-09-01

Recent studies indicate greater genetic variability among isolates of Toxoplasma gondii worldwide than previously thought. However, there is no information on genetic diversity of T. gondii from any host in Ethiopia. In the present study, genotyping was performed on viable T. gondii isolates by bioassays in mice from tissues and feces of 27 cats from Ethiopia. Viable T. gondii was isolated from hearts of 26 cats, feces alone of 1 cat, and feces and tissues of 6 cats; in total there were 33 isolates. Genotyping was performed on DNA from cell-cultured derived T. gondii tachyzoites and by using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico). Four genotypes were recognized, including ToxoDB #1 (Type II clonal, nine isolates), ToxoDB #2 (Type III, five isolates), Toxo DB #3 (Type II variant, ten isolates), and ToxoDB #20 (nine isolates). Of interest is the isolation of different genotypes from tissues and feces of two cats, suggesting re-infection or mixed strain T. gondii infection. These findings are of epidemiological significance with respect to shedding of oocysts by cats. This is the first report of genotyping of T. gondii from any host in Ethiopia. Published by Elsevier B.V.

Isolation of viable Toxoplasma gondii, molecular characterization, and seroprevalence in elk (Cervus canadensis) in Pennsylvania, USA

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Toxoplasmosis is a worldwide zoonosis. The ingestion of uncooked/undercooked meat and consumption of water contaminated with Toxoplasma gondii oocysts excreted by felids are the main modes of transmission of this parasite. Thousands of wild cervids are hunted or killed in traffic accidents yearly bu...

Seropositivity of Toxoplasma gondii in domestic donkeys (Equus asinus) and isolation of T. gondii from farm cats.

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Dubey, J P; Ness, S L; Kwok, O C H; Choudhary, S; Mittel, L D; Divers, T J

2014-01-17

Donkeys (Equus asinus) are used as both companion and working animals throughout the world and in some countries, their meat and milk are used for human consumption. Here we report the first serological survey of Toxoplasma gondii in donkeys in the United States. Serum samples from 373 donkeys from eight farms in five states were tested for T. gondii antibodies by the modified agglutination test (MAT). Twenty-four of 373 (6.4%) of donkeys were seropositive, with MAT titers ranging from 25 to ≥ 200. All seropositive donkeys were Miniature breed. Seropositivity prevalence was 7.0% in female donkeys (20/282) and 4.1% in male donkeys (4/91). No donkeys less than 24 months of age (129) were seropositive, suggesting postnatal transmission of infection. Domestic cats were present on six of the eight farms. Three cats from one farm had MAT titers of 200. Viable T. gondii was isolated from the hearts of two cats, but not from brain tissues. Genotyping of isolate DNA extracted from culture-derived tachyzoites using 10 PCR-restriction fragment length polymorphism (RFLP) markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1, L358 and Apico loci) revealed that both isolates were clonal Type II (ToxoDB PCR-RFLP genotype #1). This is the first serological survey for T. gondii in donkeys in the United States, and suggests that donkey milk and meat should be considered as a potential source for human infection. The role of barn cats in the transmission of T. gondii to donkeys on farms warrents further investigation. Copyright © 2013 Elsevier B.V. All rights reserved.

Comparative Transcriptome Analysis of Latex Reveals Molecular Mechanisms Underlying Increased Rubber Yield in Hevea brasiliensis Self-Rooting Juvenile Clones.

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Li, Hui-Liang; Guo, Dong; Zhu, Jia-Hong; Wang, Ying; Chen, Xiong-Ting; Peng, Shi-Qing

2016-01-01

Rubber tree (Hevea brasiliensis) self-rooting juvenile clones (JCs) are promising planting materials for rubber production. In a comparative trial between self-rooting JCs and donor clones (DCs), self-rooting JCs exhibited better performance in rubber yield. To study the molecular mechanism associated with higher rubber yield in self-rooting JCs, we sequenced and comparatively analyzed the latex of rubber tree self-rooting JCs and DCs at the transcriptome level. Total raw reads of 34,632,012 and 35,913,020 bp were obtained from the library of self-rooting JCs and DCs, respectively, by using Illumina HiSeq 2000 sequencing technology. De novo assemblies yielded 54689 unigenes from the library of self-rooting JCs and DCs. Among 54689 genes, 1716 genes were identified as differentially expressed between self-rooting JCs and DCs via comparative transcript profiling. Functional analysis showed that the genes related to the mass of categories were differentially enriched between the two clones. Several genes involved in carbohydrate metabolism, hormone metabolism and reactive oxygen species scavenging were up-regulated in self-rooting JCs, suggesting that the self-rooting JCs provide sufficient molecular basis for the increased rubber yielding, especially in the aspects of improved latex metabolisms and latex flow. Some genes encoding epigenetic modification enzymes were also differentially expressed between self-rooting JCs and DCs. Epigenetic modifications may lead to gene differential expression between self-rooting JCs and DCs. These data will provide new cues to understand the molecular mechanism underlying the improved rubber yield of H. brasiliensis self-rooting clones.

Loop-Mediated Isothermal Amplification-Lateral-Flow Dipstick (LAMP-LFD) to detect Toxoplasma gondii oocyst in ready-to-eat salad.

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Lalle, Marco; Possenti, Alessia; Dubey, Jitender P; Pozio, Edoardo

2018-04-01

The apicomplexan parasite Toxoplasma gondii is the causative agent of toxoplasmosis, a foodborne zoonosis with a global distribution and estimated to cause up to 20% of the total foodborne disease burden in Europe. Association between T. gondii infection and the consumption of unwashed raw fruits and vegetables contaminated with oocysts has been reported and the increasing habit to eat pre-washed ready-to-eat salads poses a new potential risk for consumers. It is therefore important to trace the occurrence of potential contamination with this parasite to guarantee the safety of ready-to-eat vegetables. Detection of T. gondii in vegetables by molecular techniques has been achieved but low sensitivity (PCR) or expensive equipments (qPCR) limit routine applicability. Here, we describe the development and validation of a sensitive and robust method relying on a LAMP assay, targeting the 529 bp locus, to detect T. gondii oocysts down to 25 oocysts/50 g in ready-to-eat baby lettuce. The LAMP has been also adapted for a faster visualization of the result by a lateral flow dipstick chromatographic detection method. Copyright © 2017 Elsevier Ltd. All rights reserved.

Prevalence of Toxoplasma gondii in Canadian market-age pigs.

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Gajadhar, A A; Aramini, J J; Tiffin, G; Bisaillon, J R

1998-08-01

During 1991 and 1992, 2,800 market-age pigs were sampled at federally inspected abattoirs from across Canada. Anti-Toxoplasma gondii IgG at titers of > or =1:32 were found in 240 pigs examined by a commercial, latex agglutination test. Seroprevalences ranged from 3.5 to 13.2% in the different regions of the country. Tissue hybridization studies using a previously developed probe demonstrated T. gondii ribosomal RNA in 9 of 36 animals, whereas mouse bioassay testing of heart muscle and diaphragm from all 2,800 pigs failed to demonstrate the presence of infective stages of T. gondii in tissues. Although serology results from this study indicated that Canadian market-age pigs are infected with T. gondii at rates similar to those reported from other parts of North America, mouse bioassay results suggested that Canadian pork products contain low levels of infective organisms. This apparent discrepancy suggests that serological evidence of T. gondii infection in pigs alone does not accurately assess the public health risks associated with consuming improperly cooked pork products.

Seroprevalence of Toxoplasma gondii infection in slaughtered pigs ...

African Journals Online (AJOL)

Toxoplasmosis is a parasitic disease/infection of medical and veterinary importance. The causative agent; Toxoplasma gondii, can infect warm blooded animals, birds as well as humans. This study was designed to determine the seroprevalence of Toxoplasma gondii infection in slaughtered pigs in Makurdi, Nigeria.

[i]Toxoplasma gondii[/i] in protected wildlife in the Tatra National Park (TANAP, Slovakia

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Ludmila Turčeková

2014-06-01

Full Text Available [i]Toxoplasma gondii[/i] is an obligatory intracellular protozoan parasite that infects a broad spectrum of warm-blooded vertebrate species. As a part of the food chain, farm animals play a significant role in transmission of [i]T. gondii [/i]to humans, while rats and mice serve as a main source of infection for free-living animals. The spread of toxoplasmosis in the human population is due to the interchange of the domestic and sylvatic cycles. During 2009–2011, a survey on toxoplasmosis distribution was conducted in wildlife of the Tatra National Park (TANAP in Slovakia. A total of 60 animals were examined. The presence of [i]T. gondii[/i] was detected by means of molecular methods based on TGR1E gene analyses. The highest prevalence was recorded in birds (40.0%, followed by carnivores (30.8% and rodents (18.2%. RFLP analyses of SAG2 locus confirmed in birds the genotype II and III, belonging to the avirulent strain; rodents exclusively had genotype I, characterised as a virulent train, and in carnivores all three genotypes were detected. These results present the first survey on the parasite’s occurrence in several species of free-living animals in the TANAP area. An epidemiological study confirmed the prevalence of 30.0%, implicitly referring to the level of environmental contamination with [i]T. gondii [/i]oocysts.

Prevalence and genetic characterization of Toxoplasma gondii in free-range chickens from grocery stores and farms in Maryland, Ohio and Massachusetts, USA.

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Ying, Yuqing; Verma, Shiv K; Kwok, Oliver C H; Alibana, Fatima; Mcleod, Rima; Su, Chunlei; Dubey, Jitender P; Pradhan, Abani K

2017-05-01

Chickens are considered important in the epidemiology of Toxoplasma gondii. Chicken hearts (n = 1185) obtained from grocery stores were tested for T. gondii infection. Antibodies to T. gondii were assayed in fluid removed from the heart cavity using the modified agglutination test (MAT) at 1:5, 1:25, and 1:100 dilutions. MAT antibodies were detected in 222 hearts at 1:5 dilution and 8 hearts at 1:25 dilution, but none were positive at 1:100 dilution. Seropositive (n = 230, 19.4%) chicken hearts were bioassayed in mice and seronegative (n = 157) chickens were bioassayed in cats. Viable T. gondii was not isolated from any hearts by bioassays in mice. The 2 cats fed 60 and 97 hearts did not excrete T. gondii oocysts. The results indicate a low prevalence of viable T. gondii in chickens from grocery stores. Molecular typing of 23 archived T. gondii strains isolated from free-range chickens from Ohio and Massachusetts using the 10 PCR-RFLP markers including SAG1, SAG2 (5'-3'SAG2 and altSAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico revealed that seven were ToxoDB PCR-RFLP genotype #1, 11 were genotype #2, one was genotype #3, three were genotype #170, and one was mixed genotype. These results indicate that the clonal genotypes #1 (type II), #2 (type III), and #3 (type II variant) are common in free-range chickens.

FastCloning: a highly simplified, purification-free, sequence- and ligation-independent PCR cloning method

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Lu Jia

2011-10-01

Full Text Available Abstract Background Although a variety of methods and expensive kits are available, molecular cloning can be a time-consuming and frustrating process. Results Here we report a highly simplified, reliable, and efficient PCR-based cloning technique to insert any DNA fragment into a plasmid vector or into a gene (cDNA in a vector at any desired position. With this method, the vector and insert are PCR amplified separately, with only 18 cycles, using a high fidelity DNA polymerase. The amplified insert has the ends with ~16-base overlapping with the ends of the amplified vector. After DpnI digestion of the mixture of the amplified vector and insert to eliminate the DNA templates used in PCR reactions, the mixture is directly transformed into competent E. coli cells to obtain the desired clones. This technique has many advantages over other cloning methods. First, it does not need gel purification of the PCR product or linearized vector. Second, there is no need of any cloning kit or specialized enzyme for cloning. Furthermore, with reduced number of PCR cycles, it also decreases the chance of random mutations. In addition, this method is highly effective and reproducible. Finally, since this cloning method is also sequence independent, we demonstrated that it can be used for chimera construction, insertion, and multiple mutations spanning a stretch of DNA up to 120 bp. Conclusion Our FastCloning technique provides a very simple, effective, reliable, and versatile tool for molecular cloning, chimera construction, insertion of any DNA sequences of interest and also for multiple mutations in a short stretch of a cDNA.

Toxoplasma gondii in wild ruminants bred in game preserves and farms with production destined for human consumption in the Czech Republic.

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Alena Lorencova

2015-08-01

Full Text Available Toxoplasma gondii is the causative agent of the most common parasitic infection in humans. Almost all warm-blooded animals, as well as humans, can act as intermediate hosts that harbour infective cysts in their tissues. Felids act as definitive hosts excreting oocysts in faeces. In humans, T. gondii can cause subclinical infection but also severe clinical disease with a wide range of symptoms, especially in immunocompromised individuals. The infection is usually asymptomatic in animals and is not recognized at either ante- or post-mortem inspection. The consumption of undercooked meat from infected animals is one of the most important routes by which the infection can be transmitted to humans. Handling of the organs and other tissues of game animals and eating their undercooked meat have been described as a risk of T. gondii infection. For diagnosis of toxoplasmosis, the combination of serological and molecular methods has been described as a suitable approach. Antibodies against T. gondii were detected in 20.8%, 50.0%, 23.1%, and 24.4% of red deer, sika deer, fallow deer and mouflons, respectively, coming from game preserves and farms in the Czech Republic. T. gondii DNA was found in the muscle tissue of red deer (8.3% and mouflons (14.6%. The lower prevalence rates based on molecular screening could be due to the random distribution and low density of cysts in tissues of infected animals. Bearing in mind the increase in the number of hunted animals and the growing trend in game consumption, it is important to educate hunters and game meat consumers about the risk of exposure to this zoonotic infection during handling and consumption of the meat.

TgICMAP1 is a novel microtubule binding protein in Toxoplasma gondii.

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Aoife T Heaslip

Full Text Available The microtubule cytoskeleton provides essential structural support for all eukaryotic cells and can be assembled into various higher order structures that perform drastically different functions. Understanding how microtubule-containing assemblies are built in a spatially and temporally controlled manner is therefore fundamental to understanding cell physiology. Toxoplasma gondii, a protozoan parasite, contains at least five distinct tubulin-containing structures, the spindle pole, centrioles, cortical microtubules, the conoid, and the intra-conoid microtubules. How these five structurally and functionally distinct sets of tubulin containing structures are constructed and maintained in the same cell is an intriguing problem. Previously, we performed a proteomic analysis of the T. gondii apical complex, a cytoskeletal complex located at the apical end of the parasite that is composed of the conoid, three ring-like structures, and the two short intra-conoid microtubules. Here we report the characterization of one of the proteins identified in that analysis, TgICMAP1. We show that TgICMAP1 is a novel microtubule binding protein that can directly bind to microtubules in vitro and stabilizes microtubules when ectopically expressed in mammalian cells. Interestingly, in T. gondii, TgICMAP1 preferentially binds to the intra-conoid microtubules, providing us the first molecular tool to investigate the intra-conoid microtubule assembly process during daughter construction.

Molecular cloning, expression analysis and sequence prediction of ...

African Journals Online (AJOL)

CCAAT/enhancer-binding protein beta as an essential transcriptional factor, regulates the differentiation of adipocytes and the deposition of fat. Herein, we cloned the whole open reading frame (ORF) of bovine C/EBPβ gene and analyzed its putative protein structures via DNA cloning and sequence analysis. Then, the ...

Isolation and characterization of a replication-competent molecular clone of an HIV-1 circulating recombinant form (CRF33_01B.

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Kok Keng Tee

Full Text Available A growing number of emerging HIV-1 recombinants classified as circulating recombinant forms (CRFs have been identified in Southeast Asia in recent years, establishing a molecular diversity of increasing complexity in the region. Here, we constructed a replication-competent HIV-1 clone for CRF33_01B (designated p05MYKL045.1, a newly identified recombinant comprised of CRF01_AE and subtype B. p05MYKL045.1 was reconstituted by cloning of the near full-length HIV-1 sequence from a newly-diagnosed individual presumably infected heterosexually in Kuala Lumpur, Malaysia. The chimeric clone, which contains the 5' LTR (long terminal repeat region of p93JP-NH1 (a previously isolated CRF01_AE infectious clone, showed robust viral replication in the human peripheral blood mononuclear cells. This clone demonstrated robust viral propagation and profound syncytium formation in CD4+, CXCR4-expressing human glioma NP-2 cells, indicating that p05MYKL045.1 is a CXCR4-using virus. Viral propagation, however, was not detected in various human T cell lines including MT-2, M8166, Sup-T1, H9, Jurkat, Molt-4 and PM1. p05MYKL045.1 appears to proliferate only in restricted host range, suggesting that unknown viral and/or cellular host factors may play a role in viral infectivity and replication in human T cell lines. Availability of a CRF33_01B molecular clone will be useful in facilitating the development of vaccine candidates that match the HIV-1 strains circulating in Southeast Asia.

Toxoplasmosis: The value of molecular methods in diagnosis compared to conventional methods

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Zineb Tlamçani

2013-06-01

Full Text Available Toxoplasmosis is a parasitic infection due to Toxoplasma gondii an obligate intracellular protozoan parasite. It is considerateone of the most common parasite worldwide. The contamination of the parasite is generally occurred via consumptionof infected food or water or, undercooked contaminated meat. Toxoplasma gondii infection may lead to seriousillness when the organism is contracted while pregnancy or when it is reactivated in immune-suppressed persons.Diagnosis of toxoplasmosis in humans is elaborated using various techniques such as detection of anti-Toxoplasmaantibodies, mouse inoculation, histological revelation of tachyzoites in tissue sections or smears of body fluid, but thedetection of Toxoplasma gondii DNA by molecular methods has revolutionized prenatal diagnosis of congenital toxoplasmosisand in immunocompromised patients. In this paper we will discuss the parasite and different methods ofdiagnosis including the usefulness of molecular methods. J Microbiol Infect Dis 2013; 3(2: 93-99Key words: Toxoplamosis, Toxoplasma gondii, diagnosis

High prevalence of Toxoplasma gondii antibodies in dogs in Veracruz, Mexico

Science.gov (United States)

Little is known concerning the prevalence of Toxoplasma gondii infection in dogs in Mexico. Here, we investigated antibodies to T. gondii and associated risk factors in 101 dogs from an animal shelter in Veracruz State, Mexico. Canine sera were assayed for T. gondii IgG antibodies by using the modif...

Toxoplasma gondii seroprevalence in dairy and beef cattle

DEFF Research Database (Denmark)

Jokelainen, Pikka; Tagel, Maarja; Motus, Kerli

2017-01-01

Toxoplasma gondii is a zoonotic protozoan parasite that thrives in Estonia. In this nationwide cross-sectional study, we tested sera from 3991 cattle, collected from 228 farms in 2012–2013, for anti-T. gondii immunoglobulin G antibodies using a commercial direct agglutination test. Titer of 100 w...

Seroprevalence of Toxoplasma gondii infection in goats from the south-west region of Poland and the detection of T. gondii DNA in goat milk.

Science.gov (United States)

Sroka, Jacek; Kusyk, Pawel; Bilska-Zajac, Ewa; Karamon, Jacek; Dutkiewicz, Jacek; Wojcik-Fatla, Angelina; Zajac, Violetta; Stojecki, Krzysztof; Rozycki, Miroslaw; Cencek, Tomasz

2017-07-11

Toxoplasma gondii (Nicolle et Manceaux, 1908) is an obligatory intracellular protozoan parasite prevalent in animals and humans worldwide having medical and veterinary importance on account of causing abortion or congenital disease in intermediate hosts, including man. Since T. gondii has already been identified in the milk of goats, Capra aegagrus hircus (Linnaeus), the possibility of acquiring infection by ingesting unpasteurised goat milk should be taken into consideration. Thus, the aim of the present study was to determine the presence of T. gondii DNA in goat milk. First, 73 goats (females) from 36 farms located in Poland were examined serologically by direct agglutination test (DAT) to estimate the T. gondii serological status. Milk samples from 60 selected lactating females were examined for the presence of T. gondii DNA by Real time PCR and nested PCR (B1 gene). To estimate the clonal type of detected T. gondii, multiplex PCR was performed using 6 markers. In DAT, positive results were found in 70% of 73 goats. Among examined 60 milk samples, 65% were positive in Real time PCR and 43% in nested PCR. It is noteworthy that 11 samples positive in PCR were collected from seronegative goats. The multilocus PCR analysis mostly revealed the occurrence of genotype III, which is relatively rare in Europe. The recorded high prevalence of anti-Toxoplasma antibodies in tested goats (70%), associated with a high prevalence of T. gondii DNA in goat milk samples (65%), indicates a potential risk of the parasite transmission through goat milk ingestion.

Comparative Transcriptome Analysis of Latex Reveals Molecular Mechanisms Underlying Increased Rubber Yield in Hevea brasiliensis Self-Rooting Juvenile Clones

OpenAIRE

Li, Hui-Liang; Guo, Dong; Zhu, Jia-Hong; Wang, Ying; Chen, Xiong-Ting; Peng, Shi-Qing

2016-01-01

Rubber tree (Hevea brasiliensis) self-rooting juvenile clones (JCs) are promising planting materials for rubber production. In a comparative trial between self-rooting JCs and donor clones (DCs), self-rooting JCs exhibited better performance in rubber yield. To study the molecular mechanism associated with higher rubber yield in self-rooting JCs, we sequenced and comparatively analyzed the latex of rubber tree self-rooting JCs and DCs at the transcriptome level. Total raw reads of 34,632,012 ...

Antiretroviral activity of protease inhibitors against Toxoplasma gondii

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Lianet Monzote

2013-02-01

Full Text Available The introduction of highly active antiretroviral therapy (HAART has caused a marked reduction in the occurrence and severity of parasitic infections, including the toxoplasmic encephalitis (TE. These changes have been attributed to the restoration of cell-mediated immunity. This study was developed to examine the activity of six antiretroviral protease inhibitors (API on Toxoplasma gondii tachyzoites. The six API showed anti-Toxoplasma activity, with IC50 value between 1.4 and 6.6 µg/mL. Further studies at the molecular level should be performed to clarify if the use of API could be beneficial or not for AIDS patients with TE.

Animal cloning: problems and prospects.

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Wells, D N

2005-04-01

An efficient animal cloning technology would provide many new opportunities for livestock agriculture, human medicine, and animal conservation. Nuclear cloning involves the production of animals that are genetically identical to the donor cells used in a technique known as nuclear transfer (NT). However, at present it is an inefficient process: in cattle, only around 6% of the embryos transferred to the reproductive tracts of recipient cows result in healthy, longterm surviving clones. Of concern are the high losses throughout gestation, during birth and in the post-natal period through to adulthood. Many of the pregnancy losses relate to failure of the placenta to develop and function correctly. Placental dysfunction may also have an adverse influence on postnatal health. These anomalies are probably due to incorrect epigenetic reprogramming of the donor genome following NT, leading to inappropriate patterns of gene expression during the development of clones. Whilst some physiological tests on surviving clones suggest normality, other reports indicate a variety of post-natal clone-associated abnormalities. This variability in outcome may reflect species-specific and/or cloning methodological differences. Importantly, to date it appears that these clone-associated phenotypes are not transmitted to offspring following sexual reproduction. This indicates that they represent epigenetic errors, rather than genetic errors, which are corrected during gametogenesis. Whilst this needs confirmation at the molecular level, it provides initial confidence in the first application of NT in agriculture, namely, the production of small numbers of cloned sires from genetically elite bulls, for natural mating, to effectively disseminate genetic gain. In addition to the animal welfare concerns with the technology, the underlying health of the animals and the consequential effect on food safety are critical aspects that require investigation to gain regulatory and consumer

Evaluation of serological and molecular tests used to identify Toxoplasma gondii infection in pregnant women attended in a public health service in São Paulo state, Brazil.

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Murata, Fernando Henrique Antunes; Ferreira, Marina Neves; Pereira-Chioccola, Vera Lucia; Spegiorin, Lígia Cosentino Junqueira Franco; Meira-Strejevitch, Cristina da Silva; Gava, Ricardo; Silveira-Carvalho, Aparecida Perpétuo; de Mattos, Luiz Carlos; Brandão de Mattos, Cinara Cássia

2017-09-01

Toxoplasmosis during pregnancy can have severe consequences. The use of sensitive and specific serological and molecular methods is extremely important for the correct diagnosis of the disease. We compared the ELISA and ELFA serological methods, conventional PCR (cPCR), Nested PCR and quantitative PCR (qPCR) in the diagnosis of Toxoplasma gondii infection in pregnant women without clinical suspicion of toxoplasmosis (G1=94) and with clinical suspicion of toxoplasmosis (G2=53). The results were compared using the Kappa index, and the sensitivity, specificity, positive predictive value and negative predictive value were calculated. The results of the serological methods showed concordance between the ELISA and ELFA methods even though ELFA identified more positive cases than ELISA. Molecular methods were discrepant with cPCR using B22/23 primers having greater sensitivity and lower specificity compared to the other molecular methods. Copyright © 2017 Elsevier Inc. All rights reserved.

Meat juice serology for Toxoplasma gondii infection in chickens

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Alice Vismarra

2016-01-01

Full Text Available Toxoplasma gondii is an important foodborne zoonosis. Free-range chickens are at particularly high risk of infection and are also excellent indicators of soil contamination by oocysts. In the present study, hearts of 77 freerange chickens were collected at slaughter. T. gondii meat juice enzyme-linked immunosorbent assay was performed with a commercial kit, following validation with positive controls, from experimentally infected chickens, and negative ones. Out of 77 samples, only 66 gave sufficient meat juice for serology. Of these, 24 (36.4% were positive for T. gondii considering the 5*standard deviation values (calculated on the optical density of negative controls, while all the samples were negative considering sample/positive% values. Parasite-specific polymerase chain reaction was carried out on all samples obtained from heart tissue and none were positive for the presence of T. gondii DNA. Results would suggest that further study on the use of meat juice with a validated serological test to detect T. gondii in chickens could lead to widespread epidemiological studies in this important intermediate host. However, sample collection and test specificity require further evaluation.

Experimental toxoplasma gondii infection in grey seals (Halichoerus grypus)

DEFF Research Database (Denmark)

Gajadhar, A. A.; Measures, L.; Forbes, L. B.

2004-01-01

Laboratory-reared animals were used to assess the susceptibility of seals (Halichoerus grypus) to Toxoplasma gondii infection. Four seals were each orally inoculated with 100 or 10,000 oocysts of T. gondii (VEG strain), and another 4 seals served as negative controls. Occasionally, mild behavioral...... changes were observed in all inoculated seals but not in control animals. A modified agglutination test revealed the presence of antibodies to T. gondii in sera collected from inoculated seals and mice inoculated as controls. No evidence of the parasite was found on an extensive histological examination...... of seal tissues, and immunohistochemical staining of tissue sections from inoculated seals revealed a single tissue cyst in only 1 seal. Control mice inoculated with 10 oocysts from the same inoculum given to seals became serologically and histologically positive for T. gondii. Cats that were fed brain...

DNA Amplification Techniques for the Detection of Toxoplasma gondii Tissue Cysts in Meat Producing Animals: A Narrative Review Article

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Farooq RIAZ

2016-12-01

Full Text Available Background: Toxoplasma gondii is an intracellular parasite, which infects one-third population of world. Humans and animals acquire infection by ingesting oocytes from feces of cats or by meat of other animals having cysts that may lead to congenital, ocular or cephalic toxoplasmosis. Either it is important to detect T. gondii from meat of food animals from retail shops or directly at slaughterhouses, which is meant for export.Methods: The current research was done without time limitation using such terms as follows: “Toxoplasma gondii”, “Meat”, “Tissue cyst”, “PCR”, “LAMP”, “Screening” and “Immunological assay” alone or in combination, in English language. The used electronic databases for searching included as follows: PubMed, Scopus, Google Scholar, Web of Science and Science Direct. The searches were limited to the published papers to English language.Results: Sensitivity of different molecular techniques for diagnosis of Toxoplasma is real-time PCR > LAMP > conventional PCR. In addition to these DNA analysis tools, bioassay in mice and cats is considered as “gold standard” to detect T. gondii. Conclusion: This review article will help the readers for grasping advantages and limitations of different diagnostic tools for screening meat samples for T. gondii. This review also makes bibliography about the type of meat sample to be processed for diagnosis and different primers or sequences to be targeted for T. gondii by number of researches for its detection from meat or tissue sample using DNA amplification techniques.

Toxoplasma gondii infection and chronic schizophrenia: is there any association?

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Salvina Maria de Campos-Carli

Full Text Available ABSTRACT Background: Toxoplasma gondii (T. gondii infection has been identified as a risk factor for schizophrenia. Objectives: Herein, we sought to evaluate the association between T. gondii infection and clinical symptoms and quality of life in patients with schizophrenia. Methods: We conducted a cross-sectional study with 48 patients with chronic schizophrenia and 40 controls. Peripheral blood was drawn, and IgM and IgG anti-T. gondii antibodies were evaluated by Enzyme-Linked Immunosorbent Assay (ELISA. Depressive, positive and negative symptoms were assessed, respectively, by the Calgary Depression Scale (CDS and the Positive and Negative Syndrome Scale (PANSS. Cognitive performance was assessed in patients by the Brazilian version of the Schizophrenia Cognition Rating Scale (SCoRS-BR. Quality of life was assessed by the Brazilian version of the Quality of Life in Schizophrenia scale (QLS-BR. Results: The prevalence and titers of T. gondii IgM and IgG antibodies did not differ between patients and controls. The positive serology for T. gondii IgG antibodies was not associated with illness symptoms, cognitive performance, depressive symptoms or quality of life. Discussion: Our findings suggest that toxoplasmosis infection is not associated with severity of symptoms, quality of life, cognitive or depressive symptoms in schizophrenia patients.

Diagnosis and isolation of Toxoplasma gondii in horses from Brazilian slaughterhouses Diagnóstico e isolamento de Toxoplasma gondii em equídeos de frigoríficos brasileiros

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Fernanda Evers

Full Text Available This study aimed to investigate anti-Toxoplasma gondii antibodies and to isolate the parasite from the brains of horses processed at slaughterhouses in Brazil. We collected brain and blood samples from 398 horses of various ages, from six Brazilian states. Serum samples were evaluated by indirect fluorescent antibody test (IFAT cut-off titre ≥ 1:64, and brains were submitted to mouse bioassay. Among the 398 horses, positivity for T. gondii was identified in 46 (11.6% by IFAT and in 14 (3.5% by mouse bioassay. In 12 of those 14 bioassays, mice were positive only by IFAT (cut-off titre ≥ 1:16, T. gondii being isolated in the remaining two. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP analysis of 18S rDNA to differentiate among T. gondii, Neospora caninum, and Sarcocystis neurona, we found that two of the 14 brains were positive for T. gondii only. For genotyping of the two isolates and the PCR-positive brain, we performed PCR-RFLP based on 13 markers, and SAG2 all samples were Toxoplasma gondii type I. Collectively, IFAT of horse sera and mouse bioassay identified positivity in 60 (15% of the samples. Our results show that some horses sent to slaughter in Brazil have been exposed to T. gondii.O objetivo do estudo foi investigar anticorpos anti-Toxoplasma gondii e isolar o parasita do cérebro de equídeos abatidos em matadouros-frigoríficos no Brasil. Colheram-se amostras de 398 cérebros e sangue de equídeos machos e fêmeas de idades variadas, provenientes de seis estados brasileiros. As amostras de soro foram avaliadas pelo teste de imunofluorescência indireta (IFI para T. gondii (ponto de corte ≥ 64, e os fragmentos de cérebros foram submetidos ao bioensaio em camundongos. Por meio da IFI, 46 (11,6% equídeos foram soropositivos. Pelo bioensaio em camundongos, 14 (3,5% cérebros de equídeos testados foram positivos. Em doze dos bioensaios, os camundongos foram positivos somente pela IFI (ponto

Toxoplasma gondii antibodies in the white stork Ciconia ciconia.

Science.gov (United States)

Andrzejewska, Izabela; Tryjanowski, Piotr; Zduniak, Piotr; Dolata, Pawel T; Ptaszyk, Jerzy; Cwiertnia, Piotr

2004-01-01

The prevalence of Toxoplasma gondii in chicks of wild birds and captive individuals was studied in the Poznań environs and in the Poznań Zoological Garden in the years 2002-2003. Bird blood was tested for T. gondii antibodies by an indirect fluorescent antibody test. T. gondii antibodies were detected from 5.8% of 205 analysed white stork chicks and 13.6% of 44 analysed adult storks in the zoo. Because toxoplasmosis is one of the more common parasitic zoonoses worldwide, we briefly discuss the potential epidemiological importance of stork toxoplasmosis to humans.

Prevalence and genetic characterization of Toxoplasma gondii in donkeys in northeastern China.

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Zhang, Xiao-Xuan; Shi, Wei; Zhang, Nian-Zhang; Shi, Kun; Li, Jian-Ming; Xu, Peng; Zhao, Quan; Du, Rui

2017-10-01

Toxolasma gondii is one of the most important obligate intracellular protozoan parasites. Recently, toxoplasmosis is of increasing concerns because T. gondii not only has a worldwide distribution but also can infect virtually all warm-blooded hosts including donkeys. However, limited information is available concerning the genetic characterization of T. gondii in donkeys in northeastern China. In this study, a total of 302 brain tissue samples collected from donkeys from Jilin (n=108) and Liaoning (n=194) provinces, northeastern China, were examined for T. gondii infection by semi-nested PCR of B1 gene. The positive samples were genotyped at 11 loci (i.e., SAG1, alternative SAG2, 5'-and 3'-SAG2, SAG3, L358, BTUB, c22-8, GRA6, c29-2, PK1 and Apico) using polymerase chain reaction-restriction fragment length polymorphism technology. Of 302 brain tissue samples, 19 (6.29%) were PCR-positive for T. gondii. The prevalence rates of T. gondii were 6.48% (7/108) and 6.19% (12/194) in Jilin Province and Liaoning Province, respectively. The prevalence of T. gondii in different season groups varied from 5.56% to 7.41%. The prevalence of T. gondii in "Cage-free" donkeys was higher than that in "Captive" donkeys. Of the 19 positive samples, only two isolates were successfully genotyped at all loci, three were genotyped at 9 loci. In total, four samples belong to ToxoDB genotype #9, one belong to ToxoDB genotype #10. This is firstly characterized the T. gondii isolates from donkeys in northeastern China. The results of the present study will improve the information of the distribution of T. gondii genotypes in China. Copyright © 2017. Published by Elsevier B.V.

Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone

International Nuclear Information System (INIS)

Adachi, A.; Gendelman, H.E.; Koenig, S.; Folks, T.; Willey, R.; Rabson, A.; Martin, M.A.

1986-01-01

The authors considered an infectious molecular clone of acquired immunodeficiency syndrome-associated retrovirus. Upon transfection, this clone directed the production of infectious virus particles in a wide variety of cells in addition to human T4 cells. The progeny, infectious virions, were synthesized in mouse, mink, monkey, and several human non-T cell lines, indicating the absence of any intracellular obstacle to viral RNA or protein production or assembly. During the course of these studies, a human colon carcinoma cell line, exquisitely sensitive to DNA transfection, was identified

Parental LTRs are important in a construct of a stable and efficient replication-competent infectious molecular clone of HIV-1 CRF08_BC.

Science.gov (United States)

Zhang, Qiwei; Zhang, Xiaomin; Wu, Hao; Seto, Donald; Zhang, Hao-Jie; Chen, Zhiwei; Wan, Chengsong; Zheng, Bo-Jian

2012-01-01

Circulating recombinant forms (CRFs) of HIV-1 have been identified in southern China in recent years. CRF08_BC is one of the most predominant subtypes circulating in China. In order to study HIV subtype biology and to provide a tool for biotechnological applications, the first full-length replication-competent infectious molecular clone harboring CRF08_BC is reported. The construction of this clone pBRGX indicates that a moderate-copy number vector is required for its amplification in E. coli. In addition, it is shown that the parental CRF08_BC LTRs are important for generating this efficient replication-competent infectious clone. These observations may aid in the construction of infectious clones from other subtypes. Both the pBRGX-derived virus and its parental isolate contain CCR5 tropism. Their full-length genomes were also sequenced, analyzed, compared and deposited in GenBank (JF719819 and JF719818, respectively). The availability of pBRGX as the first replication-competent molecular clone of CRF08_BC provides a useful tool for a wide range of studies of this newly emergent HIV subtype, including the development of HIV vaccine candidates, antiviral drug screening and drug resistance analysis.

Genotyping of Toxoplasma Gondii Isolates from Soil Samples in Tehran, Iran

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M Tavalla

2013-06-01

Full Text Available Background: The protozoan parasite Toxoplasma gondii can infect any warm blooded nucleated cells. One of the ways for human infection is ingestion of oocysts directly from soil or via infected fruits or vegetables. To survey the potential role of T. gondii oocyst in soil samples, the present study was conducted in Tehran City, Iran.Methods: A total of 150 soil samples were collected around rubbish dumps, children's play ground, parks and public places. Oocysts recovery was performed by sodium nitrate flotation method on soil samples. For molecular detection, PCR reaction targeting B1 gene was performed and then, the posi­tive results were confirmed using repetitive 529 bp DNA fragment in other PCR reaction. Finally, the positive samples were genotyped at the SAG2 locus.Results: Toxoplasma DNA was found in 13 soil samples. After genotyping and RFLP analysis in SAG2 locus, nine positive samples were revealed type III, one positive sample was type I whereas three samples revealed mixed infection (type, I & III.Conclusion: The predominant genotype in Tehran soil samples is type III.

Seroprevalence of Toxoplasma gondii in humans and pigs in North Sulawesi, Indonesia.

Science.gov (United States)

Tuda, Josef; Adiani, Sri; Ichikawa-Seki, Madoka; Umeda, Kousuke; Nishikawa, Yoshifumi

2017-10-01

Toxoplasma gondii, an intracellular protozoan parasite, is a major public health concern throughout the world. Importantly, toxoplasmosis has several adverse effects, including neurological and ocular diseases. There are currently no data on the prevalence of T. gondii infection in humans or animals in North Sulawesi, although Indonesia is known to have a high seroprevalence of this parasite. In this study, the prevalence of T. gondii was determined in samples of humans and pigs from North Sulawesi using the latex agglutination test. In total, 856 human were sampled and 58.5% of whom were positive for T. gondii. Although the antibody prevalence in male and female children aged 0-9years was 40% in both sexes, suggesting that the transmission rate of the parasite to humans is extremely high in this area. However, the overall prevalence of T. gondii in pigs was only 2.3%. Our study indicates a high incidence of T. gondii infection in humans. Therefore, a survey of the prevalence of T. gondii among different infection sources is required to determine the major risk factors for infection in North Sulawesi. Copyright © 2017 Elsevier B.V. All rights reserved.

Prevalence of Toxoplasma gondii infection in household and feral cats in Korea.

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Kim, Sung-Eon; Choi, Ran; Kang, Seung-Won; Hyun, Changbaig

2017-09-01

This study was designed to investigate the prevalence rate of Toxoplasma gondii ( T. gondii ) infection in household cats in Korea. One hundred household cats and 50 feral cats from nine of the largest cities in Korea were enrolled in this study. The tests performed in this survey was an in-house rapid screen IgG and IgM combo test, faecal PCR test for T. gondii oocysts, and an ELISA immunoassay for IgG antibodies. There were no household cats positive for T. gondii infection detected using the in-house IgG and IgM rapid screen combo test, although 6/50 and 0/50 feral cats were positive in IgG and IgM tests, respectively. This initial finding was confirmed by subsequent ELISA test for IgG antibody and PCR for T. gondii in faeces. Despite the higher prevalence rate of the disease in feral cats in Korea, we did not find any household cats that were either infected or exposed previously to T. gondii in our study population. Our study indicates that there is minimal risk of T. gondii transmission from household cats to human in Korea.

First report of Toxoplasma gondii seroprevalence in peafowls in Yunnan Province, Southwestern China

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Tian Yi-Ming

2012-09-01

Full Text Available Abstract Background Toxoplasma gondii is an intracellular protozoan parasite infecting almost all warm-blooded animals, including birds, with a worldwide distribution. Surveys of T. gondii infection in wild birds have been reported extensively in the world, but little is known of T. gondii infection in peafowls worldwide. This study was performed to determine the seroprevalence of T. gondii infection in peafowls in Yunnan Province, southwestern China. Methods Sera from 277 peafowls, including 272 blue peafowls (Pavo cristatus and 5 green peafowls (Pavo muticus originated from two geographic areas in Yunnan Province were assayed for T. gondii antibodies using the modified agglutination test (MAT. Results Specific T. gondii antibodies were detected in 35 of 277 (12.64% peafowls (MAT titer ≥ 1:5. Seropositive birds were found in both species, 33 in 272 blue peafowls and 2 in 5 green peafowls. There was no significant difference in T. gondii seroprevalence between the adolescent birds (6.74% and the adult birds (6.67% (P > 0.05. The geographical origins of peafowls was found to be highly associated with T. gondii infection in the present study, a statistically significant difference in T. gondii seropositivity was observed between peafowls from Kunming (31.08% and those from Xishuangbanna Dai Autonomous Prefecture (5.91% (OR = 10.956, 95% CI = 1.632-73.545, P = 0.014. Statistical analyses showed that there were no significant interactions between ages and geographical origins of peafowls (P > 0.05. Conclusions The results of the present survey indicated that infection of peafowls with T. gondii is widespread in Yunnan Province, which has significant public health concerns and implications for prevention and control of toxoplamosis in this province. To our knowledge, this is the first seroprevalence report of T. gondii infection in China’s southwestern Yunnan Province.

First report of Toxoplasma gondii seroprevalence in peafowls in Yunnan Province, Southwestern China.

Science.gov (United States)

Tian, Yi-Ming; Dai, Fei-Yan; Huang, Si-Yang; Deng, Zu-Hong; Duan, Gang; Zhou, Dong-Hui; Yang, Jian-Fa; Weng, Ya-Biao; Zhu, Xing-Quan; Zou, Feng-Cai

2012-09-19

Toxoplasma gondii is an intracellular protozoan parasite infecting almost all warm-blooded animals, including birds, with a worldwide distribution. Surveys of T. gondii infection in wild birds have been reported extensively in the world, but little is known of T. gondii infection in peafowls worldwide. This study was performed to determine the seroprevalence of T. gondii infection in peafowls in Yunnan Province, southwestern China. Sera from 277 peafowls, including 272 blue peafowls (Pavo cristatus) and 5 green peafowls (Pavo muticus) originated from two geographic areas in Yunnan Province were assayed for T. gondii antibodies using the modified agglutination test (MAT). Specific T. gondii antibodies were detected in 35 of 277 (12.64%) peafowls (MAT titer ≥ 1:5). Seropositive birds were found in both species, 33 in 272 blue peafowls and 2 in 5 green peafowls. There was no significant difference in T. gondii seroprevalence between the adolescent birds (6.74%) and the adult birds (6.67%) (P > 0.05). The geographical origins of peafowls was found to be highly associated with T. gondii infection in the present study, a statistically significant difference in T. gondii seropositivity was observed between peafowls from Kunming (31.08%) and those from Xishuangbanna Dai Autonomous Prefecture (5.91%) (OR = 10.956, 95% CI = 1.632-73.545, P = 0.014). Statistical analyses showed that there were no significant interactions between ages and geographical origins of peafowls (P > 0.05). The results of the present survey indicated that infection of peafowls with T. gondii is widespread in Yunnan Province, which has significant public health concerns and implications for prevention and control of toxoplamosis in this province. To our knowledge, this is the first seroprevalence report of T. gondii infection in China's southwestern Yunnan Province.

Seroepidemiology of infection with Toxoplasma gondii in healthy blood donors of Durango, Mexico

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Estrada-Martínez Sergio

2007-07-01

Full Text Available Abstract Background Toxoplasma gondii (T. gondii infection in blood donors could represent a risk for transmission in blood recipients. There is scarce information about the epidemiology of T. gondii infection in blood donors in Mexico. Therefore, we sought to determine the prevalence of T. gondii infection and associated socio-demographic and behavioral characteristics in a population of healthy blood donors of Durango City, Mexico. Methods Four hundred and thirty two blood donors in two public blood banks of Durango City, Mexico were examined for T. gondii infection between August to September 2006. Blood donors were tested for anti-T. gondii IgG and IgM antibodies by using enzyme-linked immunoassays (Diagnostic Automation Inc., Calabasas, CA, USA. Socio-demographic and behavioral characteristics from each participant were also obtained. Results Thirty two (7.4% of 432 blood donors had IgG anti-T. gondii antibodies. Eight (1.9% of them had also IgM anti-T. gondii antibodies. Multivariate analysis using logic regression showed that T. gondii infection was associated with the presence of cats at home (adjusted OR = 3.81; 95% CI: 1.45–10.01. The age group of 45–60 years showed a significantly higher frequency of T. gondii infection than the group of 25–34 years (p = 0.02. Blood donors without education had a significantly higher frequency of infection (15.8% than those with 13–19 years of education (4.5% (p = 0.04. Other characteristics of blood donors including male gender, consumption of undercooked meat or blood transfusion did not show an association with infection. Conclusion The prevalence of T. gondii infection in healthy blood donors of Durango City, Mexico is lower than those reported in blood donors of south and central Mexico, and is one of the lowest reported in blood donors worldwide. T. gondii infection in our blood donors was most likely acquired by contact with cats. Prevalence of infection increased with age and decreased

Cost-effective sequencing of full-length cDNA clones powered by a de novo-reference hybrid assembly.

Science.gov (United States)

Kuroshu, Reginaldo M; Watanabe, Junichi; Sugano, Sumio; Morishita, Shinichi; Suzuki, Yutaka; Kasahara, Masahiro

2010-05-07

Sequencing full-length cDNA clones is important to determine gene structures including alternative splice forms, and provides valuable resources for experimental analyses to reveal the biological functions of coded proteins. However, previous approaches for sequencing cDNA clones were expensive or time-consuming, and therefore, a fast and efficient sequencing approach was demanded. We developed a program, MuSICA 2, that assembles millions of short (36-nucleotide) reads collected from a single flow cell lane of Illumina Genome Analyzer to shotgun-sequence approximately 800 human full-length cDNA clones. MuSICA 2 performs a hybrid assembly in which an external de novo assembler is run first and the result is then improved by reference alignment of shotgun reads. We compared the MuSICA 2 assembly with 200 pooled full-length cDNA clones finished independently by the conventional primer-walking using Sanger sequencers. The exon-intron structure of the coding sequence was correct for more than 95% of the clones with coding sequence annotation when we excluded cDNA clones insufficiently represented in the shotgun library due to PCR failure (42 out of 200 clones excluded), and the nucleotide-level accuracy of coding sequences of those correct clones was over 99.99%. We also applied MuSICA 2 to full-length cDNA clones from Toxoplasma gondii, to confirm that its ability was competent even for non-human species. The entire sequencing and shotgun assembly takes less than 1 week and the consumables cost only approximately US$3 per clone, demonstrating a significant advantage over previous approaches.

Prevalence of Toxoplasma gondii infection in wild birds in Durango, Mexico.

Science.gov (United States)

Alvarado-Esquivel, C; Rajendran, C; Ferreira, L R; Kwok, O C H; Choudhary, S; Alvarado-Esquivel, D; Rodríguez-Peña, S; Villena, I; Dubey, J P

2011-10-01

There is a lack of information concerning the prevalence of Toxoplasma gondii infection in wild birds in Mexico. In the present study, serum samples and tissues from 653 birds from Durango State, Mexico, were evaluated for T. gondii infection. Antibodies to T. gondii (modified agglutination test, titer 1∶25 or higher) were found in 17 (2.6%) of the 653 birds, including 1 of 2 curve-billed thrashers (Toxostoma curvirostre), 2 (1 Anas platyrhynchos, 1 Anas diazi) of 4 ducks, 1 of 2 eagles (Aquila sp.), 5 (27.8%) of 18 great-tailed grackles (Quiscalus mexicanus), 7 (1.3%) of 521 rock pigeons (Columba livia), and 1 (14.3%) of 7 quail (Coturnix coturnix). The seroprevalence of T. gondii infection in birds captured in a park outside the city zoo (11.6%, 8/69) was significantly higher than that found in birds from other regions (1.5%, 9/584, OR  =  8.38; 95% CI: 2.82-24.77; P  =  0.0001). Brains and hearts of 23 birds (17 seropositive, 6 seronegative) were bioassayed in mice for the isolation of T. gondii . Viable T. gondii was isolated from 1 of 7 seropositive pigeons. The DNA obtained from the T. gondii isolate from the pigeon was genotyped using the PCR-RFLP typing using 11 markers (B1, SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) and revealed an atypical genotype. This is the first report of T. gondii infection in great-tailed grackles, the Mexican duck, and curved-billed thrashers and the first survey of wild birds in Mexico.

Cloning Expeditions: Risky but Rewarding

Science.gov (United States)

2013-01-01

In the 1980s, a good part of my laboratory was using the then-new recombinant DNA techniques to clone and characterize many important cell surface membrane proteins: GLUT1 (the red cell glucose transporter) and then GLUT2 and GLUT4, the red cell anion exchange protein (Band 3), asialoglycoprotein receptor subunits, sucrase-isomaltase, the erythropoietin receptor, and two of the subunits of the transforming growth factor β (TGF-β) receptor. These cloned genes opened many new fields of basic research, including membrane insertion and trafficking of transmembrane proteins, signal transduction by many members of the cytokine and TGF-β families of receptors, and the cellular physiology of glucose and anion transport. They also led to many insights into the molecular biology of several cancers, hematopoietic disorders, and diabetes. This work was done by an exceptional group of postdocs and students who took exceptionally large risks in developing and using novel cloning technologies. Unsurprisingly, all have gone on to become leaders in the fields of molecular cell biology and molecular medicine. PMID:24061478

MOLECULAR CLONING OF OVINE cDNA LEPTIN GENE

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CLAUDIA TEREZIA SOCOL

2008-05-01

Full Text Available An efficient bacterial transformation system suitable for cloning the coding sequence of the ovine leptin gene in E. coli DH5α host cells using the pGEMT easy vector it is described in this paper. The necessity of producing leptin is based on the fact that the role of this molecule in the animal and human organism is still unknown, leptin not existing as commercial product on the Romanian market. The results obtained in the bacterial transformation, cloning, recombinant clones selection, control of the insertion experiments and DNA computational analysis represent the first steps in further genetic engineering experiments such as production of DNA libraries, DNA sequencing, protein expression, etc., for a further contribution in elucidating the role of leptin in the animal and human organism.

Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

International Nuclear Information System (INIS)

Woon, J. S. K.; Murad, A. M. A.; Abu Bakar, F. D.

2015-01-01

A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli

Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

Energy Technology Data Exchange (ETDEWEB)

Woon, J. S. K., E-mail: jameswoon@siswa.ukm.edu.my; Murad, A. M. A., E-mail: munir@ukm.edu.my; Abu Bakar, F. D., E-mail: fabyff@ukm.edu.my [School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor (Malaysia)

2015-09-25

A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli.

Isolation, molecular cloning and expression of cellobiohydrolase B (CbhB) from Aspergillus niger in Escherichia coli

Science.gov (United States)

Woon, J. S. K.; Murad, A. M. A.; Abu Bakar, F. D.

2015-09-01

A cellobiohydrolase B (CbhB) from Aspergillus niger ATCC 10574 was cloned and expressed in E. coli. CbhB has an open reading frame of 1611 bp encoding a putative polypeptide of 536 amino acids. Analysis of the encoded polypeptide predicted a molecular mass of 56.2 kDa, a cellulose binding module (CBM) and a catalytic module. In order to obtain the mRNA of cbhB, total RNA was extracted from A. niger cells induced by 1% Avicel. First strand cDNA was synthesized from total RNA via reverse transcription. The full length cDNA of cbhB was amplified by PCR and cloned into the cloning vector, pGEM-T Easy. A comparison between genomic DNA and cDNA sequences of cbhB revealed that the gene is intronless. Upon the removal of the signal peptide, the cDNA of cbhB was cloned into the expression vector pET-32b. However, the recombinant CbhB was expressed in Escherichia coli Origami DE3 as an insoluble protein. A homology model of CbhB predicted the presence of nine disulfide bonds in the protein structure which may have contributed to the improper folding of the protein and thus, resulting in inclusion bodies in E. coli.

Outbreak of caprine abortion by Toxoplasma gondii in Midwest Brazil

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Flávio Henrique Bravim Caldeira

2011-11-01

Full Text Available An outbreak of abortion by Toxoplasma gondii in goats on a farm in the Brazilian Midwest is reported. Gross lesions were not observed in seven aborted fetuses submitted to the Veterinary Pathology Laboratory, Federal University of Mato Grosso, for necropsy investigation. The main histologic lesions were mononuclear cell pneumonia and necrotizing encephalitis in varying degrees of intensity. PCR for Brucella abortus and Neospora caninum and aerobic cultures were negative in all cases. Antibody titles against T. gondii varying from 1:1024 to 1:32.768 were detected in serum samples from four aborted goats. Nested-PCR assay for T. gondii were positive in brain samples of all cases submitted. These findings indicate that T. gondii infection should be considered in the diagnosis of abortion in goats in Midwest Brazil.

Prevalence of Toxoplasma gondii in common moles (Talpa europaea)

NARCIS (Netherlands)

Krijger, I.M.; Cornelissen, J.B.W.J.; Wisselink, H.J.; Meerburg, B.G.

2014-01-01

Background The prevalence of Toxoplasma gondii in common moles, Talpa europaea, was investigated in order to determine whether moles can serve as an indicator species for T. gondii infections in livestock. Findings In total, 86 moles were caught from 25 different sites in the Netherlands. Five

Increased apoptosis skull of pups born to Toxoplasma gondii ...

African Journals Online (AJOL)

Background: Toxoplasma gondii is an intracellular obligate protozoan parasite that infects most warm-blooded animals including humans. It can cause congenital infection with clinical symptoms ranging from mild to severe including microcephaly. At the cellular level, infection T. gondii causes apoptosis in some tissues and ...

Molecular cloning, genomic organization, and expression of a B-type (cricket-type) allatostatin preprohormone from Drosophila melanogaster

DEFF Research Database (Denmark)

Williamson, M; Lenz, C; Winther, A M

2001-01-01

and nonamidated C terminus. We have previously reported the structure of an A-type allatostatin preprohormone from the fruitfly Drosophila melanogaster. Here we describe the molecular cloning of a B-type prepro-allatostatin from Drosophila (DAP-B). DAP-B is 211 amino acid residues long and contains one copy each...

Prevalence of Toxoplasma gondii and Toxocara canis among Patients with Uveitis.

Science.gov (United States)

Lim, Su Jin; Lee, Sang Eun; Kim, Sun Hyun; Hong, Sung-Hee; You, Yong Sung; Kwon, Oh Woong; Kim, Hyeun Seung

2015-04-01

To investigate the prevalence of Toxoplasma gondii and Toxocara canis in patients with uveitis. Patients with uveitis were examined. Serum antibodies to T. gondii and T. canis were tested by using the enzyme-linked immunosorbent assay. Polymerase chain reaction (PCR) was done using blood and aqueous humor (AH). Ninety-eight patients were enrolled. Mean age was 43.5 ± 13.2 years. Six patients were seropositive for T. gondii with the following pattern: anterior uveitis, 1; posterior uveitis with retinitis, 2; pan uveitis, 2. One patient had a positive PCR result for T. gondii in AH, who showed panuveitis. Twenty-three patients were positive to serum IgG for T. canis with the following clinical manifestation: granuloma, 6; pigmented scar, 3; vitritis, 6--but none were PCR positive. T. gondii and T. canis are still important causes of uveitis. Ocular toxocariasis is not an uncommon cause of uveitis, even in adults.

THE ROLE OF HORMONES ON Toxoplasma gondii INFECTION: A SYSTEMATIC REVIEW

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María De La Luz Galván-Ramírez

2014-10-01

Full Text Available Background: Toxoplasma gondii is the causal agent of toxoplasmosis in which one third of the world’s population has been infected. In pregnant women, it may cause abortion and severe damage to the fetal central nervous system. During pregnancy, the prevalence of toxoplasmosis increases throughout the second and third quarter of gestation, simultaneously progesterone and 17β-estradiol also increase. Thus, it has been suggested that these hormones can aggravate or reduce parasite reproduction. The aim of this study was reviewing the relationship between hormones and infection caused by T. gondii in several experimental animal models and humans, focused mainly on: a congenital transmission, b parasite reproduction, c strain virulence, d levels of hormone in host induced by T. gondii infection and e participation of hormone receptors in Toxoplasma gondii infection.Are the hormones specific modulators of T. gondii infection?A systematic review methodology was used to consult several databases (Pub Med, Lilacs, Medline, Science direct, Scielo, Ebsco, Sprinker, Wiley and Google Scholar dated from September, 2013 to March, 2014. Results: 30 studies were included; eight studies in humans and 22 in animals and cell cultures. In the human studies, the most studied hormones were testosterone, progesterone, prolactin and 17-ß estradiol. Type I (RH and BK and Type II (Prugniaud, SC, ME49,T45, P78 and T38 were the most frequent experimental strains. Conclusions: Thirty-five years have passed since the first studies regarding Toxoplasma gondii infection and its relationship with hormones. This systematic review suggests that hormones modulate Toxoplasma gondii infection in different animal models. However, given that data were not comparable, further studies are required to determine the mechanism of hormone action in the Toxoplasma gondii infectious process.

Seroprevalence of Toxoplasma gondii and potential risk factors in ...

African Journals Online (AJOL)

Toxoplasma gondii infection is important in pigs and humans may get infected through the consumption of undercooked infected pork. This study conducted in Oyo state, Nigeria for 15 months (between February, 2012 and April, 2013) investigated the seroprevalence of Toxoplasma gondii infection in pigs reared on farms ...

On the determination of Toxoplasma gondii virulence in mice

Science.gov (United States)

Toxoplasma gondii is one of the most successful pathogens on earth, capable of infecting mammals and birds. Numerous papers and reports are published on isolation of T .gondii from various natural sources worldwide. The house mouse (Mus musculus) has been used as the laboratory animal model to deter...

Recombination-assisted megaprimer (RAM) cloning

Science.gov (United States)

Mathieu, Jacques; Alvarez, Emilia; Alvarez, Pedro J.J.

2014-01-01

No molecular cloning technique is considered universally reliable, and many suffer from being too laborious, complex, or expensive. Restriction-free cloning is among the simplest, most rapid, and cost-effective methods, but does not always provide successful results. We modified this method to enhance its success rate through the use of exponential amplification coupled with homologous end-joining. This new method, recombination-assisted megaprimer (RAM) cloning, significantly extends the application of restriction-free cloning, and allows efficient vector construction with much less time and effort when restriction-free cloning fails to provide satisfactory results. The following modifications were made to the protocol:•Limited number of PCR cycles for both megaprimer synthesis and the cloning reaction to reduce error propagation.•Elimination of phosphorylation and ligation steps previously reported for cloning methods that used exponential amplification, through the inclusion of a reverse primer in the cloning reaction with a 20 base pair region of homology to the forward primer.•The inclusion of 1 M betaine to enhance both reaction specificity and yield. PMID:26150930

Multiple-clone infections of Plasmodium vivax: definition of a panel of markers for molecular epidemiology.

Science.gov (United States)

de Souza, Aracele M; de Araújo, Flávia C F; Fontes, Cor J F; Carvalho, Luzia H; de Brito, Cristiana F A; de Sousa, Taís N

2015-08-25

Plasmodium vivax infections commonly contain multiple genetically distinct parasite clones. The detection of multiple-clone infections depends on several factors, such as the accuracy of the genotyping method, and the type and number of the molecular markers analysed. Characterizing the multiplicity of infection has broad implications that range from population genetic studies of the parasite to malaria treatment and control. This study compared and evaluated the efficiency of neutral and non-neutral markers that are widely used in studies of molecular epidemiology to detect the multiplicity of P. vivax infection. The performance of six markers was evaluated using 11 mixtures of DNA with well-defined proportions of two different parasite genotypes for each marker. These mixtures were generated by mixing cloned PCR products or patient-derived genomic DNA. In addition, 51 samples of natural infections from the Brazil were genotyped for all markers. The PCR-capillary electrophoresis-based method was used to permit direct comparisons among the markers. The criteria for differentiating minor peaks from artifacts were also evaluated. The analysis of DNA mixtures showed that the tandem repeat MN21 and the polymorphic blocks 2 (msp1B2) and 10 (msp1B10) of merozoite surface protein-1 allowed for the estimation of the expected ratio of both alleles in the majority of preparations. Nevertheless, msp1B2 was not able to detect the majority of multiple-clone infections in field samples; it identified only 6 % of these infections. The merozoite surface protein-3 alpha and microsatellites (PvMS6 and PvMS7) did not accurately estimate the relative clonal proportions in artificial mixtures, but the microsatellites performed well in detecting natural multiple-clone infections. Notably, the use of a less stringent criterion to score rare alleles significantly increased the sensitivity of the detection of multi-clonal infections. Depending on the type of marker used, a considerable

Seroprevalence of Anti-Toxoplasma gondii Antibodies Among Lebanese Pregnant Women.

Science.gov (United States)

Nahouli, Hasan; El Arnaout, Nour; Chalhoub, Elias; Anastadiadis, Elie; El Hajj, Hiba

2017-12-01

Toxoplasma gondii, the causative agent of toxoplasmosis, is a zoonotic obligate intracellular protozoan parasite responsible for the infection of almost one-third of the world's population. T. gondii is particularly threatening for primo-infected pregnant women and may lead, following vertical transplacental transmission, to spontaneous abortion, miscarriage, or severe manifestations in the newborn. The aim of this study was to provide an updated estimate of the seroprevalence of anti-T. gondii antibodies among a group of Lebanese pregnant women and its seroconversion rate. This is a retrospective cohort study, in which medical records of 11,000 pregnant women were screened. These women visited a private Obstetrics and Gynecology clinic located in Beirut, the capital of Lebanon, during the period of January 1994 till September 2015. Serological results of anti-T. gondii immunoglobulin G (IgG) and immunoglobulin M (IgM) results of 2456 Lebanese pregnant women who fulfilled the inclusion criteria were included in the analysis. Seropositivity and seroconversion rates for women with repeated tests were reported according to age and area of residence. The overall anti-T. gondii IgG and IgM seropositivity among 2456 Lebanese pregnant women was 82.6% and 1.8% respectively. The highest IgG seropositivity is among the age group of 35-44 years (87.81%) and at the governorate of "Mount Lebanon" (82.95%). Sixty-four seroconversions were detected and two abortions due to T. gondii infection during pregnancy were recorded. The seroprevalence of anti-T. gondii IgG among the screened pregnant women in Lebanon is the highest in the Arab region. These results highlight the importance of running a national sample survey to estimate the real potential burden of this infection and its impact on maternal and fetal health.

Ab initio multiple cloning algorithm for quantum nonadiabatic molecular dynamics

Energy Technology Data Exchange (ETDEWEB)

Makhov, Dmitry V.; Shalashilin, Dmitrii V. [Department of Chemistry, University of Leeds, Leeds LS2 9JT (United Kingdom); Glover, William J.; Martinez, Todd J. [Department of Chemistry and The PULSE Institute, Stanford University, Stanford, California 94305, USA and SLAC National Accelerator Laboratory, Menlo Park, California 94025 (United States)

2014-08-07

We present a new algorithm for ab initio quantum nonadiabatic molecular dynamics that combines the best features of ab initio Multiple Spawning (AIMS) and Multiconfigurational Ehrenfest (MCE) methods. In this new method, ab initio multiple cloning (AIMC), the individual trajectory basis functions (TBFs) follow Ehrenfest equations of motion (as in MCE). However, the basis set is expanded (as in AIMS) when these TBFs become sufficiently mixed, preventing prolonged evolution on an averaged potential energy surface. We refer to the expansion of the basis set as “cloning,” in analogy to the “spawning” procedure in AIMS. This synthesis of AIMS and MCE allows us to leverage the benefits of mean-field evolution during periods of strong nonadiabatic coupling while simultaneously avoiding mean-field artifacts in Ehrenfest dynamics. We explore the use of time-displaced basis sets, “trains,” as a means of expanding the basis set for little cost. We also introduce a new bra-ket averaged Taylor expansion (BAT) to approximate the necessary potential energy and nonadiabatic coupling matrix elements. The BAT approximation avoids the necessity of computing electronic structure information at intermediate points between TBFs, as is usually done in saddle-point approximations used in AIMS. The efficiency of AIMC is demonstrated on the nonradiative decay of the first excited state of ethylene. The AIMC method has been implemented within the AIMS-MOLPRO package, which was extended to include Ehrenfest basis functions.

Molecular cloning and characterization of Antheraea mylitta cytoplasmic polyhedrosis virus polyhedrin gene and its variant forms

International Nuclear Information System (INIS)

Sinha-Datta, Uma; Chavali, Venkata Ramana Murthy; Ghosh, Ananta K.

2005-01-01

The segments 10 (S10) of the 11 double stranded RNA genomes from Antheraea mylitta cytoplasmic polyhedrosis virus (AmCPV) encoding a novel polyhedrin polypeptide was converted to cDNA, cloned, and sequenced. Three cDNA clones consisting of 1502 (AmCPV10-1), 1120 (AmCPV10-2), and 1415 (AmCPV10-3) nucleotides encoding polyhedrin of 254, 339, and 319 amino acids with molecular masses of 29, 39, and 37 kDa, respectively, were obtained, and verified by Northern analysis. These clones showed 70-94% sequence identity among them but none with any sequences in databases. The expression of AmCPV10-1 cDNA encoded polyhedrin in Sf-9 cells was detected by immunoblot analysis and formation of polyhedra by electron microscopy, as observed in AmCPV-infected gut cells, but no expression of AmCPV10-2 or AmCPV10-3 cDNA was detected, indicating that during AmCPV replication, along with functional S10 RNA, some defective variant forms of S10 RNAs are packaged in virion particles

Detection and dissemination of Toxoplasma gondii in experimentally infected calves, a single test does not tell the whole story.

NARCIS (Netherlands)

Burrells, Alison; Taroda, Alessandra; Opsteegh, Marieke; Schares, Gereon; Benavides, Julio; Dam-Deisz, Cecile; Bartley, Paul M; Chianini, Francesca; Villena, Isabella; van der Giessen, Joke; Innes, Elisabeth A; Katzer, Frank

2018-01-01

Although the detection of Toxoplasma gondii in bovine tissues is rare, beef might be an important source of human infection. The use of molecular techniques, such as magnetic capture qPCR (MC-qPCR), in combination with the gold standard method for isolating the parasite (mouse bioassay), may

Prevalence of encysted Toxoplasma gondii in raptors from Alabama.

Science.gov (United States)

Lindsay, D S; Smith, P C; Hoerr, F J; Blagburn, B L

1993-12-01

Little is known about the prevalence of encysted Toxoplasma gondii in wild birds. We examined the hearts and breast muscles from 101 raptors for encysted T. gondii. All of the raptors had been submitted for necropsy to the State Veterinary Diagnostic Laboratory, Auburn, Alabama. Tissues were digested in acid-pepsin solution and inoculated into groups of 3-5 laboratory mice. Toxoplasma gondii was isolated from 27 of 101 (26.7%) raptors: 8 of 12 (66.7%) red-shouldered hawks (Buteo lineatus), 13 of 27 (41.1%) red-tailed hawks (Buteo jamaicensis), 1 of 4 (25%) Cooper's hawks (Accipiter cooperi), 1 of 5 (20%) great horned owls (Bubo virginianus), 4 of 15 (26.7%) barred owls (Strix varia), and 1 of 3 (33.3%) kestrels (Falco sparverius). Toxoplasma gondii was not isolated from 3 broad-winged hawks (Buteo platypterus), 3 sharp-shinned hawks (Accipiter striatus), 6 barn owls (Tyto alba), 9 screech owls (Asio otus), a Mississippi kite (Ictinia misisippiensis), 2 golden eagles (Aquila chrysaetos), a bald eagle (Haliaeetus leucocephalus), 4 ospreys (Pandion haliaetus), 4 turkey vultures (Cathartes aura), or 2 black vultures (Coragyps atratus). No significant difference (P > 0.05) in prevalence was detected based on sex using chi-square analysis. Chi-square analysis of the data demonstrated that adult raptors had encysted stages of T. gondii significantly (P < 0.05) more often than did immature raptors.

Automated cloning methods.; TOPICAL

International Nuclear Information System (INIS)

Collart, F.

2001-01-01

Argonne has developed a series of automated protocols to generate bacterial expression clones by using a robotic system designed to be used in procedures associated with molecular biology. The system provides plate storage, temperature control from 4 to 37 C at various locations, and Biomek and Multimek pipetting stations. The automated system consists of a robot that transports sources from the active station on the automation system. Protocols for the automated generation of bacterial expression clones can be grouped into three categories (Figure 1). Fragment generation protocols are initiated on day one of the expression cloning procedure and encompass those protocols involved in generating purified coding region (PCR)

Non-association between anti-Toxoplasma gondii antibodies and ABO blood group system

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ACF Rodrigues

2011-01-01

Full Text Available Toxoplasma gondii infects humans through the gastrointestinal tract (GIT, which elicits humoral immune response with specific antibodies. The expression of the ABO blood group glycoconjugates also occurs in this same system and may influence the human susceptibility of infection by T. gondii. The aim of the present study was to investigate the association between ABO blood group phenotypes and the presence of anti-T. gondii antibodies. Data - including age, results of serology tests for T. gondii infection and ABO blood group phenotypes - were assembled from the medical records of 1,006 pregnant women attended in the Base Hospital of the Medical School of São José do Rio Preto, Brazil, between 2001 and 2004. The chi-square test was used to compare the results with the level of significance set at 5%. Of the studied cases, 64.1% (645/1006 and 35.9% (391/1006 presented respectively positive and negative serology tests for anti-T. gondii antibodies. The mean age of those who tested positive was higher than those with negative serology tests (p = 0.0004. The frequencies of ABO blood group phenotypes were similar in those with and without anti-T. gondii antibodies (p = 0.35. In conclusion, the ABO blood group system is not associated with the presence or absence of anti-T. gondii antibodies.

The current status of Toxoplasma gondii infection among Egyptian rheumatoid arthritis patients

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Nagwa Mostafa El-Sayed

2016-10-01

Full Text Available Objective: To ascertain a relationship between Toxoplasma gondii (T. gondii infection and rheumatoid arthritis (RA disease among Egyptian patients. Methods: One hundred RA patients and 50 healthy subjects participated in this study. The patients were classified into three groups, namely GI, G2 and G3. Patients in G1 were recently diagnosed with RA with the disease duration of less than one year (prior treatment; G2 included RA patients receiving anti-tumor necrosis factor agents and RA patients in G3 received disease modifying anti-rheumatic drugs (methotrexate, antimalarial, corticosteroids. Serum samples of all participants were examined for the presence of anti-Toxoplasma immunoglobulin G (IgG antibodies and positive samples were further analyzed for antiToxoplasma IgM antibodies to detect the possibility of reactivation of latent toxoplasmosis. Also, the association between Toxoplasma seropositivity and clinical, laboratory and radiological features of these patients were determined. Results: There was a significantly higher percentage of T. gondii IgG positivity in RA patients (54% than in the controls (32%. At the same time, 20.40% of T. gondii IgG positive patients had anti-T. gondii IgM antibodies with a statistically significant difference as comparing to T. gondii IgG positive controls. Out of T. gondii seropositive patients, 20.37% had a lower IgG level with a mean titer of (65.3 ± 17.7 IU/mL, 46.29% had moderate level with a mean titer of (184.2 ± 60.0 IU/mL and 33.33% had higher level with a mean titer of (404.3 ± 50.0 IU/ mL. A positive correlation was found between disease activity and Toxoplasma seropositivity. T. gondii seropositive RA patients had longer disease duration, longer time morning stiffness, higher numbers of tender and swollen joints and also increase in disease severity markers (erythrocyte sedimentation rate, C-reactive protein, disease activity score 28, anti-cyclic citrullinated peptide anti

Toxoplasma gondii infection in interstate truck drivers: a case-control seroprevalence study.

Science.gov (United States)

Alvarado-Esquivel, Cosme; Pacheco-Vega, Sandy Janet; Hernández-Tinoco, Jesús; Salcedo-Jáquez, Misael; Sánchez-Anguiano, Luis Francisco; Berumen-Segovia, Luis Omar; Rábago-Sánchez, Elizabeth; Liesenfeld, Oliver

2015-02-05

Infection with Toxoplasma gondii can be acquired via the ingestion of undercooked or raw meat containing tissue cysts, or via ingestion of water contaminated with oocysts. Professional long distance truck driving may have epidemiological importance for food-borne infections since drivers eat out of home and in places where hygiene and cooking practices are uncertain. We aimed to determine whether interstate truck drivers in Durango, Mexico have an increased risk of infection with T. gondii as indicated by seropositivity; and to determine the socio-demographic, work, clinical, and behavioral characteristics associated with T. gondii seropositivity in interstate truck drivers. Through a case-control study design, 192 truck drivers and 192 controls from the general population of the same region matched by gender and age were examined with enzyme-linked immunoassays for the presence of anti-Toxoplasma IgG and IgM antibodies. Socio-demographic, work, clinical and behavioral characteristics from the truck drivers were obtained. Anti-T. gondii IgG antibodies were found in 23 (12.0%) of 192 truck drivers and in 13 (6.8%) of 192 controls (OR = 21.0; 95% CI: 1.23-358.38; P = 0.002). Anti-T. gondii IgM antibodies were found in 7 (3.6%) cases and in 7 (3.6%) controls (P = 1.00). The seroprevalence of T. gondii infection was higher in drivers with reflex impairment than in those without this impairment (4/13, 30.8% vs 19/179, 10.6%, respectively; P = 0.05), and in drivers with hearing impairment than in those without this impairment (3/7, 42.9% vs 20/185, 10.8%, respectively; P = 0.03). Multivariate analysis of work and behavioral characteristics of truck drives showed positive associations of T. gondii exposure with trips to the south of Mexico (OR = 3.11; 95% CI: 1.02-9.44; P = 0.04) and consumption of horse meat (OR = 5.18; 95% CI: 1.62-16.55; P = 0.005). Results suggest that interstate truck drivers may have an increased risk for T

Prevalence of Toxoplasma gondii infection in HIV-infected patients and food animals and direct genotyping of T. gondii isolates, Southern Ghana.

Science.gov (United States)

Pappoe, Faustina; Cheng, Weisheng; Wang, Lin; Li, Yuanling; Obiri-Yeboah, Dorcas; Nuvor, Samuel Victor; Ambachew, Henock; Hu, Xiaodong; Luo, Qingli; Chu, Deyong; Xu, Yuanhong; Shen, Jilong

2017-06-01

Toxoplasma gondii is of public health and veterinary importance causing severe diseases in immunocompromised individuals including HIV/AIDS patients and in congenital cases and animals. There is limited information on the epidemiology of T. gondii infection in humans, particularly HIV patients and food animals and the parasite genotypes in Ghana. A total of 394 HIV-infected patients from three hospitals were screened for T. gondii anti-IgG and IgM using ELISA. DNAs from blood samples of seropositve participants and 95 brain tissues of food animals were PCR assayed to detect Toxoplasma gra6. DNA positive samples were genotyped using multilocus nested polymerase chain reaction restriction fragment length polymorphism at 10 loci: sag1, alt.sag2, sag3, btub, gra6, l358, c22-8, c29-2, pk1, and apico. The overall seroprevalence was 74.37% (293/394). Toxoplasma DNAs were detected in 3.07% of the seropositive participants and 9.47% of the animals. Six of the human DNA positive samples were partly typed at sag3: 33.33, 50, and 16.67% isolates had type I, II, and III alleles, respectively. All nine isolates from food animals typed at nine loci except apico were atypical: six isolates were identical to ToxoDB #41 and #145, and one was identical to TgCkBrRj2 all identified in Brazil. The genotype of two isolates has not been reported previously and was named as TgCtGh1. T. gondii seroprevalence is high among the HIV-infected individuals with T. gondii circulating in Ghana being genetically diverse.

Toward a molecular cytogenetic map for cultivated sunflower (Helianthus annuus L.) by landed BAC/BIBAC clones.

Science.gov (United States)

Feng, Jiuhuan; Liu, Zhao; Cai, Xiwen; Jan, Chao-Chien

2013-01-01

Conventional karyotypes and various genetic linkage maps have been established in sunflower (Helianthus annuus L., 2n = 34). However, the relationship between linkage groups and individual chromosomes of sunflower remains unknown and has considerable relevance for the sunflower research community. Recently, a set of linkage group-specific bacterial /binary bacterial artificial chromosome (BAC/BIBAC) clones was identified from two complementary BAC and BIBAC libraries constructed for cultivated sunflower cv. HA89. In the present study, we used these linkage group-specific clones (~100 kb in size) as probes to in situ hybridize to HA89 mitotic chromosomes at metaphase using the BAC-fluorescence in situ hybridization (FISH) technique. Because a characteristic of the sunflower genome is the abundance of repetitive DNA sequences, a high ratio of blocking DNA to probe DNA was applied to hybridization reactions to minimize the background noise. As a result, all sunflower chromosomes were anchored by one or two BAC/BIBAC clones with specific FISH signals. FISH analysis based on tandem repetitive sequences, such as rRNA genes, has been previously reported; however, the BAC-FISH technique developed here using restriction fragment length polymorphism (RFLP)-derived BAC/BIBAC clones as probes to apply genome-wide analysis is new for sunflower. As chromosome-specific cytogenetic markers, the selected BAC/BIBAC clones that encompass the 17 linkage groups provide a valuable tool for identifying sunflower cytogenetic stocks (such as trisomics) and tracking alien chromosomes in interspecific crosses. This work also demonstrates the potential of using a large-insert DNA library for the development of molecular cytogenetic resources.

Use of filter papers to determine seroprevalence of Toxoplasma gondii among hunted ungulates

Science.gov (United States)

Toxoplasmosis is a zoonosis caused by the protozoan Toxoplasma gondii, and it is found worldwide. To determine whether ungulates are reservoirs of T. gondii in an isolated and remote region of the northeastern Peruvian Amazon, antibodies to T. gondii were determined in 5 species of ungulates by the...

Isolation and genetic characterization of Toxoplasma gondii from the gray wolf Canis lupus

Science.gov (United States)

Little is known of the genetic diversity of Toxoplasma gondii circulating in wildlife. In the present study feral gray wolf (Canis lupus) from Minnesota were examined for T. gondii infection. Antibodies to T. gondii were detected in 130 (52.4%) of 248 wolves tested by the modified agglutination test...

Seroepidemiology of Toxoplasma gondii infection in pregnant women in a public hospital in northern Mexico

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Díaz-García Juan

2006-07-01

Full Text Available Abstract Background Toxoplasma gondii (T. gondii infection in pregnant women represents a risk for congenital disease. There is scarce information about the epidemiology of T. gondii infection in pregnant women in Mexico. Therefore, we sought to determine the prevalence of T. gondii infection and associated socio-demographic, clinical and behavioural characteristics in a population of pregnant women of Durango City, Mexico. Methods Three hundred and forty three women seeking prenatal care in a public hospital of Durango City in Mexico were examined for T. gondii infection. All women were tested for anti-T. gondii IgM and IgG antibodies by using IMx Toxo IgM and IMx Toxo IgG 2.0 kits (Abbott Laboratories, Abbott Park, IL, USA, respectively. Socio-demographic, clinical and behavioural characteristics from each participant were also obtained. Results Twenty one out of the 343 (6.1% women had IgG anti-T. gondii antibodies. None of the 343 women had IgM anti-T. gondii antibodies. Multivariate analysis using logic regression showed that T. gondii infection was associated with living in a house with soil floor (adjusted OR = 7.16; 95% CI: 1.39–36.84, residing outside of Durango State (adjusted OR = 4.25; 95% CI: 1.72–10.49, and turkey meat consumption (adjusted OR = 3.85; 95% CI: 1.30–11.44. Other characteristics as cat contact, gardening, and food preferences did not show any association with T. gondii infection. Conclusion The prevalence of T. gondii infection in pregnant women of Durango City is low as compared with those reported in other regions of Mexico and the majority of other countries. Poor housing conditions as soil floors, residing in other Mexican States, and turkey meat consumption might contribute to acquire T. gondii infection.

Cloning arbuscule-related genes from mycorrhizas

DEFF Research Database (Denmark)

Burleigh, Stephen

2000-01-01

Until recently little was known about the identity of the genes expressed in the arbuscules of mycorrhizas, due in part to problems associated with cloning genes from the tissues of an obligate symbiont. However, the combination of advanced molecular techniques, innovative use of the materials...... available and fortuitous cloning has resulted in the recent identification of a number of arbuscule-related genes. This article provides a brief summary of the genes involved in arbuscule development, function and regulation, and the techniques used to study them. Molecular techniques include differential...

Molecular cloning, functional expression, and tissue distribution of a novel human gap junction-forming protein, connexin-31.9. Interaction with zona occludens protein-1

NARCIS (Netherlands)

Nielsen, Peter A; Beahm, Derek L; Giepmans, Ben N G; Baruch, Amos; Hall, James E; Kumar, Nalin M

2002-01-01

A novel human connexin gene (GJA11) was cloned from a genomic library. The open reading frame encoded a hypothetical protein of 294 amino acid residues with a predicted molecular mass of 31,933, hence referred to as connexin-31.9 (Cx31.9) or alpha 11 connexin. A clone in GenBank containing the

Molecular cloning of a functional allatostatin gut/brain receptor and an allatostatin preprohormone from the silkworm Bombyx mori

DEFF Research Database (Denmark)

Secher, Thomas; Lenz, C; Cazzamali, G

2001-01-01

in the DAR-1 and DAR-2 genes, showing that the three receptors are not only structurally but also evolutionarily related. Furthermore, we have cloned a Bombyx allatostatin preprohormone that contains eight different A-type allatostatins. Chinese hamster ovary cells permanently transfected with BAR DNA react......The cockroach-type or A-type allatostatins are inhibitory insect neuropeptides with the C-terminal sequence Tyr/Phe-X-Phe-Gly-Leu-NH(2). Here, we have cloned an A-type allatostatin receptor from the silkworm Bombyx mori (BAR). BAR is 361 amino acid residues long, has seven transmembrane domains....... Northern blots and quantitative reverse transcriptase-polymerase chain reaction of different larval tissues show that BAR mRNA is mainly expressed in the gut and to a much lesser extent in the brain. To our knowledge, this is the first report on the molecular cloning and functional expression of an insect...

Anti-gluten immune response following Toxoplasma gondii infection in mice.

Directory of Open Access Journals (Sweden)

Emily G Severance

Full Text Available Gluten sensitivity may affect disease pathogenesis in a subset of individuals who have schizophrenia, bipolar disorder or autism. Exposure to Toxoplasma gondii is a known risk factor for the development of schizophrenia, presumably through a direct pathological effect of the parasite on brain and behavior. A co-association of antibodies to wheat gluten and to T. gondii in individuals with schizophrenia was recently uncovered, suggesting a coordinated gastrointestinal means by which T. gondii and dietary gluten might generate an immune response. Here, we evaluated the connection between these infectious- and food-based antigens in mouse models. BALB/c mice receiving a standard wheat-based rodent chow were infected with T. gondii via intraperitoneal, peroral and prenatal exposure methods. Significant increases in the levels of anti-gluten IgG were documented in all infected mice and in offspring from chronically infected dams compared to uninfected controls (repetitive measures ANOVAs, two-tailed t-tests, all p≤0.00001. Activation of the complement system accompanied this immune response (p≤0.002-0.00001. Perorally-infected females showed higher levels of anti-gluten IgG than males (p≤0.009 indicating that T. gondii-generated gastrointestinal infection led to a significant anti-gluten immune response in a sex-dependent manner. These findings support a gastrointestinal basis by which two risk factors for schizophrenia, T. gondii infection and sensitivity to dietary gluten, might be connected to produce the immune activation that is becoming an increasingly recognized pathology of psychiatric disorders.

Urine sample used for detection of toxoplasma gondii infection by loop-mediated isothermal amplification (LAMP).

Science.gov (United States)

Hu, Xin; Pan, Chang-Wang; Li, Ya-Fei; Wang, Han; Tan, Feng

2012-02-01

In this study, a loop-mediated isothermal amplification (LAMP) assay was established to detect Toxoplasma gondii DNA in mice infected with T. gondii PRU strain. This LAMP assay was based on the sequence of highly repetitive B1 gene. The detection limit of T. gondii LAMP assay was 1 pg of T. gondii DNA, which was evaluated using 10-fold serially diluted DNA of cultured parasites. The LAMP assay was also highly specific for T. gondii and able to detect T. gondii DNA in urine of mice treated with dexamethasone at 90 day post infection (p.i.), although this assay could not detect the DNA in mice urine 2-6 days p.i. These results demonstrated that LAMP is effective for evaluation of therapy effectiveness for T. gondii infection. The established LAMP assay may represent a useful and practical tool for the routine diagnosis and therapeutic evaluation of human toxoplasmosis.

ELISA-seroprevalence of Toxoplasma gondii in draught horses in Greater Cairo, Egypt.

Science.gov (United States)

Haridy, Fouad M; Shoukry, Nahla M; Hassan, Aly Awad; Morsy, Tosson A

2009-12-01

Toxoplasma gondii is one of the important zoonotic parasites of worldwide. In this paper the seroprevalence of T. gondii in draught horses (3-15 years) including 90 males and 10 females in the first half of the year 2009 was studied. The result showed that the overall ELISA-T. gondii antibodies were 25% of the horses in Greater Cairo, 50% (females) and 22.2% (males).

Alterações cognitivas em ratos infectados com Toxoplasma gondii

OpenAIRE

Maia, Raquel da Silveira

2012-01-01

O Toxoplasma gondii é um protozoário parasito que induz alterações comportamentais em roedores. O objetivo do presente estudo foi avaliar o efeito da infecção pelo T.gondii durante a fase crônica na memória operacional e na impulsividade de roedores, bem como o efeito de antipsicóticos em reverter as eventuais alterações comportamentais decorrentes da infecção. Ratos wistar fêmeas (n=40) foram infectadas com 25 cistos da cepa ME-49 do T. gondii, após 4 meses os animais foram submetidos aos te...

Lack of association between Toxoplasma gondii infection and occupational exposure to animals

Science.gov (United States)

Pacheco-Vega, Sandy Janet; Hernández-Tinoco, Jesús; Saldaña-Simental, Diana Elizabeth; Sánchez-Anguiano, Luis Francisco; Salcedo-Jáquez, Misael; Ramos-Nevárez, Agar; Liesenfeld, Oliver; Márquez-Conde, José Ángel; Cerrillo-Soto, Sandra Margarita; Martínez-Ramírez, Lucio; Guido-Arreola, Carlos Alberto

2014-01-01

The association of infection with Toxoplasma gondii and occupational exposure to animals has been scantly determined. We performed a case-control study with 200 subjects from Durango Province, Mexico, occupationally exposed to animals and 200 age- and gender-matched subjects without this occupation. Sera from all participants were analyzed for anti-T. gondii IgG and IgM antibodies using enzyme-linked immunoassays. The association of seroprevalence with sociodemographic, work, clinical, and behavioral characteristics in cases was determined. Cases and controls had similar frequencies of anti-T. gondii IgG antibodies (12/200: 6.0% and 11/200: 5.5%, respectively) (OR = 3.0; 95% CI: 0.12–73.64; P = 1.0). The frequency of sera with high (>150 IU/ml) levels of anti-T. gondii IgG antibodies was comparable among cases and controls (P = 0.61). Seroprevalence of anti-T. gondii IgM antibodies was similar in cases (4, 2.0%) than in controls (4, 2.0%) (P = 1.0). Multivariate analysis showed that seropositivity was associated with eating while working (OR = 7.14; 95% CI: 1.91–26.72; P = 0.003) and consumption of duck meat (OR = 5.43; 95% CI: 1.43–20.54; P = 0.01). No association between seropositivity to T. gondii and occupational exposure to animals was found. However, risk factors for infection found should be taken into account to reduce the exposure to T. gondii. PMID:25544890

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)

Toxoplasma gondii IgG antibodies in HIV/AIDS patients attending ...

African Journals Online (AJOL)

Background: Toxoplasmosis is caused by Toxoplasma gondii, a parasite that gradually evolved to be the most opportunistic parasite that complicates the course of HIV/AIDS in developing countries. Aim: This study was undertaken to investigate the presence of Toxoplasma gondii IgG antibodies in HIVinfected patients ...

DNA cloning: a practical approach. Volume 1

Energy Technology Data Exchange (ETDEWEB)

Glover, D M [ed.

1985-01-01

This book is written for the advanced molecular biologist who needs a detailed discussion of cloning technology. Topics of discussion include: genomic library cloning (size of a genomic library, screening methods, chromosome walking, host cell genetics, and general features of bacteriophage Iambda); use of gt10 and gt11 cDNA lambda vectors and general cDNA cloning; RNase H-Pol I cDNA synthesis; method of detecting fusion proteins produced in bacteria; pEMBL family of double-stranded plasmid vectors that can be used to generate single strands; Escherichia coli transformation; production of mutations in cloned sequences; and cloning in gram negative bacteria.

Synergy of mIL-21 and mIL-15 in enhancing DNA vaccine efficacy against acute and chronic Toxoplasma gondii infection in mice.

Science.gov (United States)

Li, Zhong-Yuan; Chen, Jia; Petersen, Eskild; Zhou, Dong-Hui; Huang, Si-Yang; Song, Hui-Qun; Zhu, Xing-Quan

2014-05-23

The synergistic protective efficacy of murine interleukin 21 (mIL-21) and mIL-15 administrated with DNA vaccine against acute and chronic Toxoplasma gondii infection in mice was investigated using T. gondii MIC8 (TgMIC8) as a model. We cloned mIL-21 and mIL-15 from splenic tissues of Kunming mice, and constructed eukaryotic plasmid pVAX/mIL-15, pVAX/mIL-21, and pVAX/mIL-21/mIL-15, respectively. After immunizing with pVAX/TgMIC8 in the presence or absence of these cytokines, immune responses were analyzed using lymphoproliferative assay, cytokine and serum antibody measurements, flow cytometric surface markers on lymphocytes and protection against acute and chronic T. gondii infection. Mice receiving pVAX/TgMIC8 alone developed a strong humoral responses and Th1 type cellular immune responses, and showed an increase of CD4+ and CD8+ T cells compared with all the controls. Adding pVAX/mIL-21 to pVAX/TgMIC8 compared to pVAX/TgMIC8 resulted in only a slight increase in humoral and cellular immune responses, and this immune response was lower than that induced by the pVAX/mIL-15 combined with pVAX/TgMIC8. Co-administration of pVAX/mIL-21/mIL-15 combined with pVAX/TgMIC8 elicited the strongest humoral and cellular immune responses among all the groups, leading to significantly increased survival time against acute infection and the significant reduction of tissue cysts, compared to all the controls. Synergy of mIL-21 and mIL-15 can facilitate specific humoral as well as cellular immune responses elicited by DNA vaccine against acute and chronic T. gondii infection in mice. Copyright © 2014 Elsevier Ltd. All rights reserved.

Molecular cloning and chromosome mapping of the human gene encoding protein phosphotyrosyl phosphatase 1B

International Nuclear Information System (INIS)

Brown-Shimer, S.; Johnson, K.A.; Bruskin, A.; Green, N.R.; Hill, D.E.; Lawrence, J.B.; Johnson, C.

1990-01-01

The inactivation of growth suppressor genes appears to play a major role in the malignant process. To assess whether protein phosphotyrosyl phosphatases function as growth suppressors, the authors have isolated a cDNA clone encoding human protein phosphotyrosyl phosphatase 1B for structural and functional characterization. The translation product deduced from the 1,305-nucleotide open reading frame predicts a protein containing 435 amino acids and having a molecular mass of 49,966 Da. The amino-terminal 321 amino acids deduced from the cDNA sequence are identical to the empirically determined sequence of protein phosphotyrosyl phosphatase 1B. A genomic clone has been isolated and used in an in situ hybridization to banded metaphase chromosomes to determine that the gene encoding protein phosphotyrosyl phosphatase 1B maps as a single-copy gene to the long arm of chromosome 20 in the region q13.1-q13.2

Hepatoprotective activity of Thymus vulgaris extract against Toxoplasma gondii infection

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Nagwa Mostafa El-Sayed

2017-05-01

Full Text Available Objective: To evaluate the hepatoprotective activity of Thymus vulgaris (T. vulgaris extract against Toxoplasma gondii (T. gondii infection in experimentally infected mice. Methods: Sixty mice were divided into six groups (Group I–Group VI. Group I was normal control (non-infected, non-treated; Group II was non-infected and treated with T. vulgaris extract (500 mg/kg; Group III was T. gondii infected-non-immunosuppressed control; Group IV consisted of infected immunosuppressed mice; Group V was infected and treated with T. vulgaris extract; Group VI consisted of infected immunosuppressed mice treated with T. vulgaris extract. Hepatoprotective effect of T. vulgaris extract was evaluated by histopathological examination of tissue sections stained with hematoxylin and eosin, determination of liver function parameters (alanine aminotransaminase, aspartate aminotransaminase and alkaline phosphates, total bilirubin, total protein concentrations and assessment of hepatocytes genotoxicity by comet assay.Antigenotoxic effect of T. vulgaris was assessed by several comet assay parameters that were provided by the image analysis software, including % tailed cells, % of DNA in the tail, tail length, and tail moment. Results: Treatment with T. vulgaris in both Groups V and VI improved T. gondii induced pathological lesions in the infected liver that regressed to near the normal picture especially in Group V. Also, it restored the altered values of liver function parameters near to the normal levels significantly (P < 0.05 compared with Groups III and IV respectively. Regarding comet assay parameters, all of them were significantly increased (P < 0.05 after T. gondii infection (Group III and reached the greatest values in infected immunosuppressed group (Group IV compared to the normal controls (Group I. With treatment by T. vulgaris in Groups V and VI, there was a significant decrease (P < 0.05 in all values compared to Groups III and V respectively. The

Serological evidence of Toxoplasma gondii infection in captive marine mammals in Mexico.

Science.gov (United States)

Alvarado-Esquivel, C; Sánchez-Okrucky, R; Dubey, J P

2012-03-23

Toxoplasma gondii infection in marine mammals is important because they are considered as a sentinel for contamination of seas with T. gondii oocysts, and toxoplasmosis causes mortality in these animals, particularly sea otters. Serological evidence of T. gondii infection was determined in 75 captive marine mammals from four facilities in southern and central geographical regions in Mexico using the modified agglutination test (MAT). Antibodies (MAT, 1:25 or higher) to T. gondii were found in 55 (87.3%) of 63 Atlantic bottlenose dolphins (Tursiops truncatus truncatus), 3 of 3 Pacific bottlenose dolphins (Tursiops truncatus gillii), 2 of 4 California sea lions (Zalophus californianus), but not in 3 West Indian manatees (Trichechus manatus), and 2 Patagonian sea lions (Otaria flavescens). Seropositive marine mammals were found in all 4 (100%) facilities sampled. All marine mammals were healthy and there has not been any case of clinical toxoplasmosis in the facilities sampled for at least the last 15 years. The seroprevalence of T. gondii infection in marine mammals of the same species did not vary significantly with respect to sex and age. This is the first report on the detection of antibodies to T. gondii in marine mammals in Mexico. Published by Elsevier B.V.

Toxoplasma gondii Infection Is Associated with Mitochondrial Dysfunction in-Vitro

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Genevieve Syn

2017-12-01

Full Text Available Upon invasion of host cells, the ubiquitous pathogen Toxoplasma gondii manipulates several host processes, including re-organization of host organelles, to create a replicative niche. Host mitochondrial association to T. gondii parasitophorous vacuoles is rapid and has roles in modulating host immune responses. Here gene expression profiling of T. gondii infected cells reveals enrichment of genes involved in oxidative phosphorylation (OXPHOS and mitochondrial dysfunction 6 h post-infection. We identified 11 hub genes (HIF-1α, CASP8, FN1, POU5F1, CD44, ISG15, HNRNPA1, MDM2, RPL35, VHL, and NUPR1 and 10 predicted upstream regulators, including 4 endogenous regulators RICTOR, KDM5A, RB1, and D-glucose. We characterized a number of mitochondrial parameters in T. gondii infected human foreskin fibroblast cells over a 36 h time-course. In addition to the usual rapid recruitment and apparent enlargement of mitochondria around the parasitophorous vacuole we observed fragmented host mitochondria in infected cells, not linked to cellular apoptosis, from 24 h post-infection. An increase in mitochondrial superoxide levels in T. gondii infected cells was observed that required active parasite invasion and peaked at 30 h post-infection. Measurement of OXPHOS proteins showed decreased expression of Complex IV in infected cells at 24 h post-infection, followed by decreased expression of Complexes I and II at 36 h post-infection. No change occurred in Complex V. No difference in host mitochondrial membrane potential between infected and mock-infected cells was observed at any time. Our results show perturbation of host mitochondrial function following T. gondii infection that likely impacts on pathogenesis of disease.

Seroprevalence and risk factors for Toxoplasma gondii infection in domestic cats in The Netherlands

NARCIS (Netherlands)

Opsteegh, M.; Haveman, R.; Swart, A.; Mensink-Beerepoot, M.E.; Hofhuis, A.; Langelaar, M.F.M.; van der Giessen, J.W.B.

2012-01-01

Cats, as definitive hosts, play an important role in the transmission of Toxoplasma gondii. To determine the seroprevalence and risk factors for T. gondii infection in Dutch domestic cats, serum samples of 450 cats were tested for T. gondii antibodies by indirect ELISA. Binary mixture analysis was

A novel multifunctional oligonucleotide microarray for Toxoplasma gondii

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Chen Feng

2010-10-01

Full Text Available Abstract Background Microarrays are invaluable tools for genome interrogation, SNP detection, and expression analysis, among other applications. Such broad capabilities would be of value to many pathogen research communities, although the development and use of genome-scale microarrays is often a costly undertaking. Therefore, effective methods for reducing unnecessary probes while maintaining or expanding functionality would be relevant to many investigators. Results Taking advantage of available genome sequences and annotation for Toxoplasma gondii (a pathogenic parasite responsible for illness in immunocompromised individuals and Plasmodium falciparum (a related parasite responsible for severe human malaria, we designed a single oligonucleotide microarray capable of supporting a wide range of applications at relatively low cost, including genome-wide expression profiling for Toxoplasma, and single-nucleotide polymorphism (SNP-based genotyping of both T. gondii and P. falciparum. Expression profiling of the three clonotypic lineages dominating T. gondii populations in North America and Europe provides a first comprehensive view of the parasite transcriptome, revealing that ~49% of all annotated genes are expressed in parasite tachyzoites (the acutely lytic stage responsible for pathogenesis and 26% of genes are differentially expressed among strains. A novel design utilizing few probes provided high confidence genotyping, used here to resolve recombination points in the clonal progeny of sexual crosses. Recent sequencing of additional T. gondii isolates identifies >620 K new SNPs, including ~11 K that intersect with expression profiling probes, yielding additional markers for genotyping studies, and further validating the utility of a combined expression profiling/genotyping array design. Additional applications facilitating SNP and transcript discovery, alternative statistical methods for quantifying gene expression, etc. are also pursued at

Brain cancer mortality rates increase with Toxoplasma gondii seroprevalence in France

Science.gov (United States)

Vittecoq, Marion; Elguero, Eric; Lafferty, Kevin D.; Roche, Benjamin; Brodeur, Jacques; Gauthier-Clerc, Michel; Missé, Dorothée; Thomas, Frédéric

2012-01-01

The incidence of adult brain cancer was previously shown to be higher in countries where the parasite Toxoplasma gondii is common, suggesting that this brain protozoan could potentially increase the risk of tumor formation. Using countries as replicates has, however, several potential confounding factors, particularly because detection rates vary with country wealth. Using an independent dataset entirely within France, we further establish the significance of the association between T. gondii and brain cancer and find additional demographic resolution. In adult age classes 55 years and older, regional mortality rates due to brain cancer correlated positively with the local seroprevalence of T. gondii. This effect was particularly strong for men. While this novel evidence of a significant statistical association between T. gondii infection and brain cancer does not demonstrate causation, these results suggest that investigations at the scale of the individual are merited.

Seroprevalence of Toxoplasma gondii in Danish farmed mink (Mustela vison S.)

DEFF Research Database (Denmark)

Henriksen, P; Dietz, H. H.; Uttenthal, Åse

1994-01-01

One hundred and ninety-five mink sera randomly selected from 17 Danish mink farms were evaluated for the presence of Toxoplasma gondii antibodies in the latex agglutination test. Six (3%) sera contained T. gondii antibodies in titres of 1:64 or more. The estimated 3% prevalence means that 300 000...... mink out of a total mink population of ten million might be infected with Toxoplasma gondii. This large number of possible sero-positive mink in Denmark indicates that there exists a potential risk of acquiring toxoplasmosis by pelting mink....

Sequence variation in TgROP7 gene among Toxoplasma gondii ...

African Journals Online (AJOL)

Yomi

2012-03-27

Mar 27, 2012 ... Toxoplasma gondii can infect a wide range of hosts including mammals and birds, causing toxoplasmosis which is one of the most common parasitic zoonoses worldwide. The present study examined sequence variation in rhoptry 7 (ROP7) gene among different T. gondii isolates from different hosts and ...

Detection of Toxolasma gondii in captive wild felids.

Science.gov (United States)

Buddhirongawatr, Ruangrat; Tungsudjai, Siriporn; Chaichoune, Kridsada; Sangloung, Charoonluk; Tantawiwattananon, Nitipan; Phonaknguen, Rassameepen; Sukthana, Yaowalark

2006-01-01

Toxoplasma gondii can infect all species of warm-blooded animals, including humans, and causes serious diseases in immunocompromized hosts. Live tachyzoites derived from serial passage in HeLa culture were used in the Sabin-Feldman dye test for detection of Toxoplasma gondii antibody in serum samples of 21 captive wild felids including one fishing cat (Prion nailurus viverrina), one leopard (Panthera pardus), two flat-headed cats (Prion nailurus planiceps), 6 tigers (Panthera tigris), two leopard cats (Felis bengalensis), two clouded leopards (Felis nebulosa), 3 pumas (Puma concolor), and 4 jungle cats (Felis chaus). Antibodies to Toxoplasma gondii were founded in 9 of 21 felids (42.8%). This study revealed that cell culture-derived tachyzoites can be used successfully as a source of live organisms in a gold standard Sabin-Feldman dye test, which is simpler, cheaper and less ethically sensitive than in vivo inoculation.

Isolation of Toxoplasma gondii from horse meat in Egypt.

Science.gov (United States)

Shaapan, R M; Ghazy, A A

2007-01-01

Portions of heart, liver, skeletal and diaphragmatic muscles obtained from 150 slaughtered horses at Giza-Zoo abattoir were used for bioassays in mice and cats. T. gondii tachyzoites were isolated successfully from the peritoneal exudates of the inoculated mice 6-8 days post inoculation with pooled horse tissues. Whereas, T. gondii tissue cysts containing bradyzoites were detected in the impression smears of mice brain on the 45th days or more post infection. The oocysts were detected in feces of cats 3-6 days post feeding on horse tissues containing tissue cysts. The oocysts became sporulated within 3-5 days in 2.5% Potassium dichromate. A total of 79 out of 150 horse meat samples were found to be infected with an incidence rate of 52.6 %. This is the first trial for isolation of T. gondii infective stages from horses in Egypt. Moreover, this study pointed out to the high infection rate of T. gondii in horse meat which may be considered as an important source of infection to wild zoo-animals in Egypt and humans in some countries if consumed raw or insufficiently cooked.

"Latent" infection with Toxoplasma gondii: association with trait aggression and impulsivity in healthy adults.

Science.gov (United States)

Cook, Thomas B; Brenner, Lisa A; Cloninger, C Robert; Langenberg, Patricia; Igbide, Ajirioghene; Giegling, Ina; Hartmann, Annette M; Konte, Bettina; Friedl, Marion; Brundin, Lena; Groer, Maureen W; Can, Adem; Rujescu, Dan; Postolache, Teodor T

2015-01-01

Latent chronic infection with Toxoplasma gondii (T. gondii), a common neurotropic pathogen, has been previously linked with suicidal self-directed violence (SSDV). We sought to determine if latent infection with T. gondii is associated with trait aggression and impulsivity, intermediate phenotypes for suicidal behavior, in psychiatrically healthy adults. Traits of aggression and impulsivity were analyzed in relationship to IgG antibody seropositivity for T. gondii and two other latent neurotropic infections, herpes simplex virus 1 (HSV1) and cytomegalovirus (CMV). One thousand community-residing adults residing in the Munich metropolitan area with no Axis I or II conditions by SCID for DSM-IV (510 men, 490 women, mean age 53.6 ± 15.8, range 20-74). Plasma samples were tested for IgG antibodies to T. gondii, HSV-1 and CMV by ELISA. Self-reported ratings of trait aggression scores (Questionnaire for Measuring Factors of Aggression [FAF]) and trait impulsivity (Sensation-Seeking Scale-V [SSS-V]) were analyzed using linear multivariate methods. T. gondii IgG seropositivity was significantly associated with higher trait reactive aggression scores among women (p impulsive sensation-seeking (SSS-V Disinhibition) among younger men (p impulsivity, personality traits considered as endophenotypes for SSDV, are associated with latent T. gondii infection in a gender and age-specific manner, and could be further investigated as prognostic and treatment targets in T. gondii-positive individuals at risk for SSDV. Published by Elsevier Ltd.

Human uroporphyrinogen III synthase: Molecular cloning, nucleotide sequence, and expression of a full-length cDNA

International Nuclear Information System (INIS)

Tsai, Shihfeng; Bishop, D.F.; Desnick, R.J.

1988-01-01

Uroporphyrinogen III synthase, the fourth enzyme in the heme biosynthetic pathway, is responsible for conversion of the linear tetrapyrrole, hydroxymethylbilane, to the cyclic tetrapyrrole, uroporphyrinogen III. The deficient activity of URO-synthase is the enzymatic defect in the autosomal recessive disorder congenital erythropoietic porphyria. To facilitate the isolation of a full-length cDNA for human URO-synthase, the human erythrocyte enzyme was purified to homogeneity and 81 nonoverlapping amino acids were determined by microsequencing the N terminus and four tryptic peptides. Two synthetic oligonucleotide mixtures were used to screen 1.2 x 10 6 recombinants from a human adult liver cDNA library. Eight clones were positive with both oligonucleotide mixtures. Of these, dideoxy sequencing of the 1.3 kilobase insert from clone pUROS-2 revealed 5' and 3' untranslated sequences of 196 and 284 base pairs, respectively, and an open reading frame of 798 base pairs encoding a protein of 265 amino acids with a predicted molecular mass of 28,607 Da. The isolation and expression of this full-length cDNA for human URO-synthase should facilitate studies of the structure, organization, and chromosomal localization of this heme biosynthetic gene as well as the characterization of the molecular lesions causing congenital erythropoietic porphyria

Concentration and retention of Toxoplasma gondii surrogates from seawater by red abalone (Haliotis rufescens)

Science.gov (United States)

Schott, Kristen C; Krusor, Colin; Tinker, M. Tim; Moore, James G.; Conrad, Patricia A.; Shapiro, Karen

2016-01-01

Small marine snails and abalone have been identified as high- and low-risk prey items, respectively, for exposure of threatened southern sea otters to Toxoplasma gondii, a zoonotic parasite that can cause fatal encephalitis in animals and humans. While recent work has characterized snails as paratenic hosts for T. gondii, the ability of abalone to vector the parasite has not been evaluated. To further elucidate why abalone predation may be protective against T. gondii exposure, this study aimed to determine whether: (1) abalone are physiologically capable of acquiring T. gondii; and (2) abalone and snails differ in their ability to concentrate and retain the parasite. Abalone were exposed to T. gondii surrogate microspheres for 24 h, and fecal samples were examined for 2 weeks following exposure. Concentration of surrogates was 2–3 orders of magnitude greater in abalone feces than in the spiked seawater, and excretion of surrogates continued for 14 days post-exposure. These results indicate that, physiologically, abalone and snails can equally vector T. gondii as paratenic hosts. Reduced risk of T. gondii infection in abalone-specializing otters may therefore result from abalone's high nutritional value, which implies otters must consume fewer animals to meet their caloric needs.

Concentration and retention of Toxoplasma gondii surrogates from seawater by red abalone (Haliotis rufescens).

Science.gov (United States)

Schott, Kristen C; Krusor, Colin; Tinker, M Tim; Moore, James; Conrad, Patricia A; Shapiro, Karen

2016-11-01

Small marine snails and abalone have been identified as high- and low-risk prey items, respectively, for exposure of threatened southern sea otters to Toxoplasma gondii, a zoonotic parasite that can cause fatal encephalitis in animals and humans. While recent work has characterized snails as paratenic hosts for T. gondii, the ability of abalone to vector the parasite has not been evaluated. To further elucidate why abalone predation may be protective against T. gondii exposure, this study aimed to determine whether: (1) abalone are physiologically capable of acquiring T. gondii; and (2) abalone and snails differ in their ability to concentrate and retain the parasite. Abalone were exposed to T. gondii surrogate microspheres for 24 h, and fecal samples were examined for 2 weeks following exposure. Concentration of surrogates was 2-3 orders of magnitude greater in abalone feces than in the spiked seawater, and excretion of surrogates continued for 14 days post-exposure. These results indicate that, physiologically, abalone and snails can equally vector T. gondii as paratenic hosts. Reduced risk of T. gondii infection in abalone-specializing otters may therefore result from abalone's high nutritional value, which implies otters must consume fewer animals to meet their caloric needs.

Toxoplasma gondii in horse meat intended for human consumption in Romania.

Science.gov (United States)

Paştiu, Anamaria Ioana; Györke, Adriana; Kalmár, Zsuzsa; Bolfă, Pompei; Rosenthal, Benjamin Martin; Oltean, Miruna; Villena, Isabelle; Spînu, Marina; Cozma, Vasile

2015-09-15

The prevalence of Toxoplasma gondii, an economically important zoonotic protozoan, was investigated in horses slaughtered for export and human consumption in the North of Romania. Pairs of samples, sera and heart tissues, were collected from 82 slaughtered horses. Examination of horse sera by ELISA at a dilution of 1:10, and by modified agglutination test (MAT) at a dilution of 1:6, revealed that 32 (39%) and 31(37.8%) horses, respectively, had antibodies against T. gondii. Using polymerase chain reaction (PCR) analysis, T. gondii DNA was not found in any heart sample collected from horses. By bioassay in mice, we obtained viable isolates of T. gondii from two of ten horses determined to be strongly positive by serological assay/ELISA. The prevalence estimated in horses highlighted the potential risk for human contamination by consumption of raw or undercooked meat. Copyright © 2015 Elsevier B.V. All rights reserved.

Toxoplasma gondii, Neospora caninum, Sarcocystis neurona, and Sarcocystis canis-like infections in marine mammals

Science.gov (United States)

Dubey, J.P.; Zarnke, R.; Thomas, N.J.; Wong, S.K.; Vanbonn, W.; Briggs, M.; Davis, J.W.; Ewing, R.; Mense, M.; Kwok, O.C.H.; Romand, S.; Thulliez, P.

2003-01-01

Toxoplasma gondii, Neospora caninum, Sarcocystis neurona, and S. canis are related protozoans that can cause mortality in many species of domestic and wild animals. Recently, T. gondii and S. neurona were recognized to cause encephalitis in marine mammals. As yet, there is no report of natural exposure of N. caninum in marine mammals. In the present study, antibodies to T. gondii and N. caninum were assayed in sera of several species of marine mammals. For T. gondii, sera were diluted 1:25, 1:50, and 1:500 and assayed in the T. gondii modified agglutination test (MAT). Antibodies (MAT a?Y1:25) to T. gondii were found in 89 of 115 (77%) dead, and 18 of 30 (60%) apparently healthy sea otters (Enhydra lutris), 51 of 311 (16%) Pacific harbor seals (Phoca vitulina), 19 of 45 (42%) sea lions (Zalophus californianus), 5 of 32 (16%) ringed seals (Phoca hispida), 4 of 8 (50%) bearded seals (Erignathus barbatus), 1 of 9 (11.1%) spotted seals (Phoca largha), 138 of 141 (98%) Atlantic bottlenose dolphins (Tursiops truncatus), and 3 of 53 (6%) walruses (Odobenus rosmarus). For N. caninum, sera were diluted 1:40, 1:80, 1:160, and 1:320 and examined with the Neospora agglutination test (NAT) using mouse-derived tachyzoites. NAT antibodies were found in 3 of 53 (6%) walruses, 28 of 145 (19%) sea otters, 11 of 311 (3.5%) harbor seals, 1 of 27 (3.7%) sea lions, 4 of 32 (12.5%) ringed seals, 1 of 8 (12.5%) bearded seals, and 43 of 47 (91%) bottlenose dolphins. To our knowledge, this is the first report of N. caninum antibodies in any marine mammal, and the first report of T. gondii antibodies in walruses and in ringed, bearded, spotted, and ribbon seals. Current information on T. gondii-like and Sarcocystis-like infections in marine mammals is reviewed. New cases of clinical S. canis and T. gondii infections are also reported in sea lions, and T. gondii infection in an Antillean manatee (Trichechus manatus manatus).

Antibody Detection, Isolation, Genotyping, and Virulence of Toxoplasma gondii in Captive Felids from China

Directory of Open Access Journals (Sweden)

Yu-Rong Yang

2017-07-01

Full Text Available The felids are the only definitive hosts of Toxoplasma gondii, which could excrete oocysts into the environment and provide an infection source for toxoplasmosis in various warm-blooded animal species, particularly the captive felids that live close to human communities. The infection rate of the captive felids is a perfect standard in detecting the presence of Toxoplasma gondii oocysts in the environment. In this study, sera or tissue samples from zoo (1 young tiger, 2 adult tigers, 6 young lions, farm (10 masked palm civets, and pet hospital (28 cats from Henan Province (China were collected. The sera (n = 47 were tested for immunoglobulin G (IgG antibodies against T. gondii by using modified agglutination test (MAT, whereas the hearts tissue (n = 40 were bioassayed in mice to isolate T. gondii strains. The genotype was distinguished by using PCR-RFLP of 10 loci (SAG1, SAG2, SAG3, GRA6, BTUB, L358, c22-8, PK1, c29-2, and Apico. The detection rate for the T. gondii antibody in captive felids was 21.3% (10/47. One viable T. gondii strain (TgCatCHn4 was obtained from a cat heart tissue, and its genotype was ToxoDB#9. The oocysts of ToxoDB#9 were collected from a T. gondii-free cat. The virulence of TgCatCHn4 was low and no cysts were detected in the brain of mice at 60 days post-inoculation. The finding of the present study suggested a widespread exposure of T. gondii for felids in Henan Province of central China, particularly those from the zoological gardens and homes. ToxoDB#9 was the predominant strain in China. Preventive measures against T. gondii oocyst contamination of various components of the environment should thus be implemented, including providing pre-frozen meat, well-cooked cat food, cleaned fruits and vegetables, monitoring birds and rodents, inactive T. gondii oocysts in felids feces, and proper hygiene.

Seroprevalence of Toxoplasma gondii infection in feral cats in Qatar.

Science.gov (United States)

Boughattas, Sonia; Behnke, Jerzy; Sharma, Aarti; Abu-Madi, Marawan

2017-01-18

Cats are essential in the life cycle of Toxoplasma gondii as they can shed the environmentally resistant oocysts after acquiring infection. Human populations living in cities with high densities of feral cats are therefore likely to be at risk of infection. The current study is the first to estimate the seroprevalence of T. gondii in the feral cat population in Qatar. We investigated the seroprevalence of T. gondii among 495 adult cats from urban and suburban districts in Qatar. Using results from the Modified Agglutination Test, we fitted statistical models with host sex, area and season as explanatory factors and seropositivity as the outcome. The analysis revealed an overall seroprevalence of 82%. Seroprevalence was significantly higher in the summer season (P = 0.006). No significant difference was detected (P > 0.05) between seroprevalence in female and male cats and in cats from urban and suburban districts of Qatar. Despite the seasonal difference, the observed seroprevalence of T. gondii suggests high environmental contamination throughout the year, with some female cats generating more intense responses compared to males. Both findings merit further investigations.

Global Analysis of Palmitoylated Proteins in Toxoplasma gondii.

Science.gov (United States)

Foe, Ian T; Child, Matthew A; Majmudar, Jaimeen D; Krishnamurthy, Shruthi; van der Linden, Wouter A; Ward, Gary E; Martin, Brent R; Bogyo, Matthew

2015-10-14

Post-translational modifications (PTMs) such as palmitoylation are critical for the lytic cycle of the protozoan parasite Toxoplasma gondii. While palmitoylation is involved in invasion, motility, and cell morphology, the proteins that utilize this PTM remain largely unknown. Using a chemical proteomic approach, we report a comprehensive analysis of palmitoylated proteins in T. gondii, identifying a total of 282 proteins, including cytosolic, membrane-associated, and transmembrane proteins. From this large set of palmitoylated targets, we validate palmitoylation of proteins involved in motility (myosin light chain 1, myosin A), cell morphology (PhIL1), and host cell invasion (apical membrane antigen 1, AMA1). Further studies reveal that blocking AMA1 palmitoylation enhances the release of AMA1 and other invasion-related proteins from apical secretory organelles, suggesting a previously unrecognized role for AMA1. These findings suggest that palmitoylation is ubiquitous throughout the T. gondii proteome and reveal insights into the biology of this important human pathogen. Copyright © 2015 Elsevier Inc. All rights reserved.

Chronic Toxoplasma gondii in Nurr1-null heterozygous mice exacerbates elevated open field activity.

Science.gov (United States)

Eells, Jeffrey B; Varela-Stokes, Andrea; Guo-Ross, Shirley X; Kummari, Evangel; Smith, Holly M; Cox, Erin; Lindsay, David S

2015-01-01

Latent infection with Toxoplasma gondii is common in humans (approximately 30% of the global population) and is a significant risk factor for schizophrenia. Since prevalence of T. gondii infection is far greater than prevalence of schizophrenia (0.5-1%), genetic risk factors are likely also necessary to contribute to schizophrenia. To test this concept in an animal model, Nurr1-null heterozygous (+/-) mice and wild-type (+/+) mice were evaluate using an emergence test, activity in an open field and with a novel object, response to bobcat urine and prepulse inhibition of the acoustic startle response (PPI) prior to and 6 weeks after infection with T. gondii. In the emergence test, T. gondii infection significantly decreased the amount of time spent in the cylinder. Toxoplasma gondii infection significantly elevated open field activity in both +/+ and +/- mice but this increase was significantly exacerbated in +/- mice. T. gondii infection reduced PPI in male +/- mice but this was not statistically significant. Aversion to bobcat urine was abolished by T. gondii infection in +/+ mice. In female +/- mice, aversion to bobcat urine remained after T. gondii infection while the male +/- mice showed no aversion to bobcat urine. Antibody titers of infected mice were a critical variable associated with changes in open field activity, such that an inverted U shaped relationship existed between antibody titers and the percent change in open field activity with a significant increase in activity at low and medium antibody titers but no effect at high antibody titers. These data demonstrate that the Nurr1 +/- genotype predisposes mice to T. gondii-induced alterations in behaviors that involve dopamine neurotransmission and are associated with symptoms of schizophrenia. We propose that these alterations in murine behavior were due to further exacerbation of the altered dopamine neurotransmission in Nurr1 +/- mice.

Chronic Toxoplasma gondii in Nurr1-null heterozygous mice exacerbates elevated open field activity.

Directory of Open Access Journals (Sweden)

Jeffrey B Eells

Full Text Available Latent infection with Toxoplasma gondii is common in humans (approximately 30% of the global population and is a significant risk factor for schizophrenia. Since prevalence of T. gondii infection is far greater than prevalence of schizophrenia (0.5-1%, genetic risk factors are likely also necessary to contribute to schizophrenia. To test this concept in an animal model, Nurr1-null heterozygous (+/- mice and wild-type (+/+ mice were evaluate using an emergence test, activity in an open field and with a novel object, response to bobcat urine and prepulse inhibition of the acoustic startle response (PPI prior to and 6 weeks after infection with T. gondii. In the emergence test, T. gondii infection significantly decreased the amount of time spent in the cylinder. Toxoplasma gondii infection significantly elevated open field activity in both +/+ and +/- mice but this increase was significantly exacerbated in +/- mice. T. gondii infection reduced PPI in male +/- mice but this was not statistically significant. Aversion to bobcat urine was abolished by T. gondii infection in +/+ mice. In female +/- mice, aversion to bobcat urine remained after T. gondii infection while the male +/- mice showed no aversion to bobcat urine. Antibody titers of infected mice were a critical variable associated with changes in open field activity, such that an inverted U shaped relationship existed between antibody titers and the percent change in open field activity with a significant increase in activity at low and medium antibody titers but no effect at high antibody titers. These data demonstrate that the Nurr1 +/- genotype predisposes mice to T. gondii-induced alterations in behaviors that involve dopamine neurotransmission and are associated with symptoms of schizophrenia. We propose that these alterations in murine behavior were due to further exacerbation of the altered dopamine neurotransmission in Nurr1 +/- mice.

Epidemiological aspects of Toxoplasma gondii infection in riverside communities in the Southern Brazilian Amazon

Directory of Open Access Journals (Sweden)

Sérgio Neto Vitaliano

2015-06-01

Full Text Available INTRODUCTION: Toxoplasma gondii infection is widely prevalent in humans and other animals worldwide. Information on the prevalence of T. gondii infection is scarce in some regions of Brazil, including riverside communities along the Amazon River basin. M METHODS: The prevalence of T. gondii in 231 people, aged 1-85 years, who were living in four riverside communities along the Purus River, Lábrea, State of Amazonas, Brazil, was determined. Antibodies against T. gondii were assayed using a commercial enzyme-linked immunosorbent assay (ELISA kit. The hearts and brains of 50 chickens, which were raised free-range in the communities, were pooled according to the community of origin and bioassayed in mice. The isolates were genotyped using polymorphisms at 12 nuclear markers (SAG1, 5' and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. RESULTS: The overall seroprevalence of T. gondii was 56.7% (131/231. IgG antibodies were presented by 117 (89.3% and IgM by 14 (10.7% of the 131 positive individuals. No association between age group and gender with prevalence was observed (chi-square test, p > 0.05; however, the comparison between localities showed that the seroprevalence of T. gondii was significantly lower among the individuals living in the Boca do Ituxi (p < 0.05 community. Five isolates of T. gondii were obtained in the mouse bioassay, and genotyping revealed two complete genotypes that had not been described previously and three mixed isolates. CONCLUSIONS: These results support previous findings that T. gondii population genetics are highly diverse in Brazil and that T. gondii infection is active in these riverside communities.

Molecular cloning and expression analysis of a cDNAs encoding androgenic gland hormone precursors from two Porcellionidae species, Porcellio scaber and P. dilatatus

OpenAIRE

Ohira, Tsuyoshi; Hasegawa, Yuriko; Okuno, Atsuro; Nagasawa, Hiromichi

2003-01-01

Male sexual characteristics in Crustacea are induced by androgenic gland hormone (AGH), which is produced by the male-specific androgenic gland. Recently, AGH in the terrestrial isopod Armadillidium vulgare was characterized and its cDNA cloned, the first example in which the structure of AGH was elucidated. We report here the molecular cloning of cDNAs encoding AGH precursors from two additional terrestrial isopods, Porcellio scaber and P. dilatatus. cDNA fragments encoding Porcellio scaber ...

Molecular cloning of growth hormone encoding cDNA of Indian

Indian Academy of Sciences (India)

A modified rapid amplification of cDNA ends (RACE) strategy has been developed for cloning highly conserved cDNA sequences. Using this modified method, the growth hormone (GH) encoding cDNA sequences of Labeo rohita, Cirrhina mrigala and Catla catla have been cloned, characterized and overexpressed in ...

Molecular cloning of lupin leghemoglobin cDNA

DEFF Research Database (Denmark)

Konieczny, A; Jensen, E O; Marcker, K A

1987-01-01

Poly(A)+ RNA isolated from root nodules of yellow lupin (Lupinus luteus, var. Ventus) has been used as a template for the construction of a cDNA library. The ds cDNA was synthesized and inserted into the Hind III site of plasmid pBR 322 using synthetic Hind III linkers. Clones containing sequences...... specific for nodules were selected by differential colony hybridization using 32P-labeled cDNA synthesized either from nodule poly(A)+ RNA or from poly(A)+ RNA of uninfected root as probes. Among the recombinant plasmids, the cDNA gene for leghemoglobin was identified. The protein structure derived from...... its nucleotide sequence was consistent with known amino acid sequence of lupin Lb II. The cloned lupin Lb cDNA hybridized to poly(A)+ RNA from nodules only, which is in accordance with the general concept, that leghemoglobin is expressed exclusively in nodules. Udgivelsesdato: 1987-null...

Detection of infection with toxoplasma Gondii in manatees (Trichechus inunguis) of the peruvian amazon

International Nuclear Information System (INIS)

Mathews Delgado, Patrick; Sanchez Perea, Nofre; Mathews Delgado, John Paul; Biffi Garcia, Claudia; Malheiros, Antonio Francisco; Garcia Davila, Carmen Rosa

2013-01-01

The Amazonian manatee (trichechus inunguis) is an aquatic mammal that inhabits freshwater environments and is endemic to the amazon basin. The presence of Toxoplasma gondii antibodies was investigated in 19 manatees in one rescue unit in the northern region of Peru. Antibodies to T. gondii were detected in 12 (63.2 %) of 19 animals by using the modified agglutination test (titer, 1:25), and no association between sex and age of the animals and the presence of T. gondii antibodies was observed (p < 0.05). the results suggest a contamination by T. gondii oocysts in the aquatic environment where these animals live.

Toxoplasma gondii abortion storm in sheep on a Texas farm and isolation of mouse virulent atypical genotype T. gondii from an aborted lamb from a chronically infected ewe

Science.gov (United States)

Sheep are commonly infected with the protozoan parasite, Toxoplasma gondii. Infection may cause early embryonic death and resorption, fetal death and mummification, abortion, stillbirth, and neonatal death. Most sheep acquire T. gondii infection after birth. Recent studies reported that repeat ovine...

Toxoplasma gondii Infection in Seagull Chicks Is Related to the Consumption of Freshwater Food Resources

Science.gov (United States)

Cabezón, Oscar; Cerdà-Cuéllar, Marta; Morera, Virginia; García-Bocanegra, Ignacio; González-Solís, Jacob; Napp, Sebastian; Ribas, Maria P.; Blanch-Lázaro, Berta; Fernández-Aguilar, Xavier; Antilles, Noelia; López-Soria, Sergio; Lorca-Oró, Cristina; Dubey, Jitender P.; Almería, Sonia

2016-01-01

Understanding the spread of Toxoplasma gondii (T. gondii) in wild birds, particularly in those with opportunistic feeding behavior, is of interest for elucidating the epidemiological involvement of these birds in the maintenance and dissemination of the parasite. Overall, from 2009 to 2011, we collected sera from 525 seagull chicks (Yellow-legged gull (Larus michahellis) and Audouin’s gull (L. audouinii)) from 6 breeding colonies in Spain and tested them using the modified agglutination test (MAT) for the presence of antibodies against T. gondii. Chick age was estimated from bill length. Main food source of seagull chicks was evaluated using stable isotope analyses from growing scapular feathers. Overall T. gondii seroprevalence was 21.0% (IC95% 17.5–24.4). A generalized linear mixed-effects model indicated that year (2009) and food source (freshwater) were risk factors associated to the individual risk of infection by T. gondii, while age (days) was close to significance. Freshwater food origin was related to the highest seroprevalence levels, followed by marine origin, supporting freshwater and sewages as important routes of dispersion of T. gondii. Year differences could indicate fluctuating rates of exposure of seagull chicks to T. gondii. Age ranged from 4 to 30 days and seropositivity tended to increase with age (P = 0.07), supporting that seropositivity is related to T. gondii infection rather than to maternal transfer of antibodies, which in gulls is known to sharply decrease with chick age. This study is the first to report T. gondii antibodies in Yellow-legged and Audouin’s gulls, thereby extending the range of intermediate hosts for this parasite and underscoring the complexity of its epidemiology. PMID:26974667

Dual Sarcocystis neurona and Toxoplasma gondii infection in a northern sea otter from Washington state, USA

Science.gov (United States)

Lindsay, D.S.; Thomas, N.J.; Rosypal, A.C.; Dubey, J.P.

2001-01-01

Dual Sarcocystis neurona and Toxoplasma gondii infection was observed in a Northern sea otter from Washington, USA. The animal was found stranded, convulsed, and died shortly thereafter. Encephalitis caused by both S. neurona and T. gondii was demonstrated in histological sections of brain. Immunohistochemical examination of sections with S. neurona specific antisera demonstrated developmental stages that divided by endopolygeny and produced numerous merozoites. PCR of brain tissue from the sea otter using primer pairs JNB33/JNB54 resulted in amplification of a 1100 bp product. This PCR product was cut in to 884 and 216 bp products by Dra I but was not cut by Hinf I indicating that it was S. neurona [J. Parasitol. 85 (1999) 221]. No PCR product was detected in the brain of a sea otter which had no lesions of encephalitis. Examination of brain sections using T. gondii specific antisera demonstrated tachyzoites and tissue cysts of T. gondii. The lesions induced by T. gondii suggested that the sea otter was suffering from reactivated toxoplasmosis. T. gondii was isolated in mice inoculated with brain tissue. A cat that was fed infected mouse brain tissue excreted T. gondii oocysts which were infective for mice. This is apparently the first report of dual S. neurona and T. gondii in a marine mammal.

Toxoplasma gondii seroprevalence and association with equine protozoal myeloencephalitis: A case-control study of Californian horses.

Science.gov (United States)

James, K E; Smith, W A; Packham, A E; Conrad, P A; Pusterla, N

2017-06-01

While toxoplasmosis is not commonly considered a clinical disease of equines, previous seroprevalence studies have reported differing background rates of Toxoplasma gondii infection in horses globally. The objective of this study was to evaluate possible associations between T. gondii seroprevalence and clinical signs of equine protozoal myeloencephalitis (EPM) in horses. Using a case-control study design, 720 Californian horses with neurologic signs compatible with EPM were compared to healthy, non-neurologic horses for the presence of T. gondii antibodies (using indirect fluorescent antibody tests [IFAT]). Toxoplasma gondii seroprevalence among cases and controls was determined at standard serum cut-offs: 40, 80, 160, 320, and 640. At a T. gondii titre cut-off of 320, horses with clinical signs compatible with EPM had 3.55 times the odds of a seropositive test compared to those without clinical signs (Pequines. Serologic testing of cerebrospinal fluid and isolation of T. gondii in EPM suspect cases should be considered. Future studies investigating the relationship between T. gondii and EPM are warranted. Copyright © 2017 Elsevier Ltd. All rights reserved.

A Toxoplasma gondii protein with homology to intracellular type Na+/H+ exchangers is important for osmoregulation and invasion

International Nuclear Information System (INIS)

Francia, Maria E.; Wicher, Sarah; Pace, Douglas A.; Sullivan, Jack; Moreno, Silvia N.J.; Arrizabalaga, Gustavo

2011-01-01

The obligate intracellular parasite Toxoplasma gondii is exposed to a variety of physiological conditions while propagating in an infected organism. The mechanisms by which Toxoplasma overcomes these dramatic changes in its environment are not known. In yeast and plants, ion detoxification and osmotic regulation are controlled by vacuolar compartments. A novel compartment named the plant-like vacuole or vacuolar compartment (PLV/VAC) has recently been described in T.gondii, which could potentially protect extracellular tachyzoites against salt and other ionic stresses. Here, we report the molecular characterization of the vacuolar type Na + /H + exchanger in T. gondii, TgNHE3, and its co-localization with the PLV/VAC proton-pyrophosphatase (TgVP1). We have created a TgNHE3 knockout strain, which is more sensitive to hyperosmotic shock and toxic levels of sodium, possesses a higher intracellular Ca 2+ concentration [Ca 2+ ] i , and exhibits a reduced host invasion efficiency. The defect in invasion correlates with a measurable reduction in the secretion of the adhesin TgMIC2. Overall, our results suggest that the PLV/VAC has functions analogous to those of the vacuolar compartments of plants and yeasts, providing the parasite with a mechanism to resist ionic fluctuations and, potentially, regulate protein trafficking.

Molecular cloning and characterization of recA-like gene from Schizosaccharomyces pombe

International Nuclear Information System (INIS)

Lee, J.S.; Kang, J.K.; Yoon, S.M.; Park, Y.; Yang, Y.K.; Kim, S.W.; Park, J.K.; Park, J.G.; Hong, S.H.; Park, S.D.

1996-01-01

We have previously purified and characterized a RecA-like protein from Schizosaccharomyces pombe (S. pombe). In the present study, we have cloned a gene encoding the RecA-like protein. The S. pombe recA-like gene was isolated by immunological screening of the expression library of S. pombe using anti-Escherichia coli (E. coli) RecA antibody as a probe. From 10(6) plaques screened, 6 putative clones were finally isolated. Five of the clones screened contained the same kinds of DNA inserts, as determined by crosshybridization analysis. Among the clones, TC-2 was selected for further studies. The pGEM3Zf(-)Delta 17 vector harboring the 4.3 kb DNA insert of TC-2 clone was capable of producing abeta-gal/RecA-like fusion protein, suggesting that the cloned gene encodes the RecA-like protein of S. pombe. It was also revealed by Southern hybridization analysis that the same DNA sequence as the cloned recA-like gene is located within the S. pombe chromosomal DNA. In addition, the cloned recA-like gene was transcribed into a 3.0 kb RNA transcript, as judged by Northern blot analysis. The level of the RNA transcript of recA-like gene was increased approximately 1.6 to 2.4-fold upon treatment with DNA damaging agents such as ultraviolet (UV)-light, methyl methanesulfonate (MMS), and mitomycin-C (MMC). This data suggests that the cloned S. pombe recA-like gene is slightly inducible to DNAdamage as in E. coli recA gene. These results suggest that an inducible repair mechanism analogous to that of E. coli may exist in fission yeast S. pombe

Molecular cloning and expression analysis of annexin A2 gene in sika deer antler tip

Directory of Open Access Journals (Sweden)

Yanling Xia

2018-04-01

Full Text Available Objective Molecular cloning and bioinformatics analysis of annexin A2 (ANXA2 gene in sika deer antler tip were conducted. The role of ANXA2 gene in the growth and development of the antler were analyzed initially. Methods The reverse transcriptase polymerase chain reaction (RT-PCR was used to clone the cDNA sequence of the ANXA2 gene from antler tip of sika deer (Cervus Nippon hortulorum and the bioinformatics methods were applied to analyze the amino acid sequence of Anxa2 protein. The mRNA expression levels of the ANXA2 gene in different growth stages were examined by real time reverse transcriptase polymerase chain reaction (real time RT-PCR. Results The nucleotide sequence analysis revealed an open reading frame of 1,020 bp encoding 339 amino acids long protein of calculated molecular weight 38.6 kDa and isoelectric point 6.09. Homologous sequence alignment and phylogenetic analysis indicated that the Anxa2 mature protein of sika deer had the closest genetic distance with Cervus elaphus and Bos mutus. Real time RT-PCR results showed that the gene had differential expression levels in different growth stages, and the expression level of the ANXA2 gene was the highest at metaphase (rapid growing period. Conclusion ANXA2 gene may promote the cell proliferation, and the finding suggested Anxa2 as an important candidate for regulating the growth and development of deer antler.

Molecular Cloning, Expression and Characterization of Pectin Methylesterase (CtPME) from Clostridium thermocellum.

Science.gov (United States)

Rajulapati, Vikky; Goyal, Arun

2017-05-01

Many phytopathogenic micro-organisms such as bacteria and fungi produce pectin methylesterases (PME) during plant invasion. Plants and insects also produce PME to degrade plant cell wall. In the present study, a thermostable pectin methylesterase (CtPME) from Clostridium thermocellum belonging to family 8 carbohydrate esterase (CE8) was cloned, expressed and purified. The amino acid sequence of CtPME exhibited similarity with pectin methylesterase from Erwinia chrysanthemi with 38% identity. The gene encoding CtPME was cloned into pET28a(+) vector and expressed using Escherichia coli BL21(DE3) cells. The recombinant CtPME expressed as a soluble protein and exhibited a single band of molecular mass approximately 35.2 kDa on SDS-PAGE gels. The molecular mass, 35.5 kDa of the enzyme, was also confirmed by MALDI-TOF MS analysis. Notably, highest protein concentration (11.4 mg/mL) of CtPME was achieved in auto-induction medium, as compared with LB medium (1.5 mg/mL). CtPME showed maximum activity (18.1 U/mg) against citrus pectin with >85% methyl esterification. The optimum pH and temperature for activity of CtPME were 8.5 and 50 °C, respectively. The enzyme was stable in pH range 8.0-9.0 and thermostable between 45 and 70 °C. CtPME activity was increased by 40% by 5 mM Ca 2+ or Mg 2+ ions. Protein melting curve of CtPME gave a peak at 80 °C. The peak was shifted to 85 °C in the presence of 5 mM Ca 2+ ions, and the addition of 5 mM EDTA shifted back the melting peak to 80 °C. CtPME can be potentially used in food and textile industry applications.

Molecular cloning, sequence analysis and phylogeny of first caudata g-type lysozyme in axolotl (Ambystoma mexicanum).

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Yu, Haining; Gao, Jiuxiang; Lu, Yiling; Guang, Huijuan; Cai, Shasha; Zhang, Songyan; Wang, Yipeng

2013-11-01

Lysozymes are key proteins that play important roles in innate immune defense in many animal phyla by breaking down the bacterial cell-walls. In this study, we report the molecular cloning, sequence analysis and phylogeny of the first caudate amphibian g-lysozyme: a full-length spleen cDNA library from axolotl (Ambystoma mexicanum). A goose-type (g-lysozyme) EST was identified and the full-length cDNA was obtained using RACE-PCR. The axolotl g-lysozyme sequence represents an open reading frame for a putative signal peptide and the mature protein composed of 184 amino acids. The calculated molecular mass and the theoretical isoelectric point (pl) of this mature protein are 21523.0 Da and 4.37, respectively. Expression of g-lysozyme mRNA is predominantly found in skin, with lower levels in spleen, liver, muscle, and lung. Phylogenetic analysis revealed that caudate amphibian g-lysozyme had distinct evolution pattern for being juxtaposed with not only anura amphibian, but also with the fish, bird and mammal. Although the first complete cDNA sequence for caudate amphibian g-lysozyme is reported in the present study, clones encoding axolotl's other functional immune molecules in the full-length cDNA library will have to be further sequenced to gain insight into the fundamental aspects of antibacterial mechanisms in caudate.

Stillbirth history and Toxoplasma gondii infection in women attending public health centers in a northern Mexican City

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Alvarado-Esquivel, C.; Pacheco-Vega, S. J.; Salcedo-Jaquez, M.; Sánchez-Anguiano, L. F.; Hernández-Tinoco, J.; Rábago-Sánchez, E.; Centeno-Tinoco, M. M.; Flores-Garcia, I. D.; Ramos-Nevarez, A.; Cerrillo-Soto, S. M.; Guido-Arreola, C. A.; Beristain-García, I.; Liesenfeld, O.; Berumen-Segovia, L. O.; Saenz-Soto, L.; Sifuentes-Álvarez, A.

2015-01-01

Through a cross-sectional study design, 150 women attending public health centers with a history of stillbirths were examined for anti-Toxoplasma gondii IgG and IgM antibodies in Durango City, Mexico. Bivariate and multivariate analyses were used to assess the association of T. gondii seropositivity with the characteristics of the women with stillbirth history. Of the 150 women (mean age: 32.09 ± 9.16 years) studied, 14 (9.3%) had anti-T. gondii IgG antibodies and six (42.9%) of them were also positive for anti-T. gondii IgM antibodies. Multivariate analysis showed that T. gondii seropositivity was associated with high frequency (4–7 days a week) of eating meat (OR = 5.52; 95% CI: 1.48–20.59; P = 0.01), history of lymphadenopathy (OR = 4.52; 95% CI: 1.14–17.82; P = 0.03), and history of surgery (OR = 8.68; 95% CI: 1.04–72.15; P = 0.04). This is the first study on the seroepidemiology of T. gondii infection in women with a history of stillbirths in Mexico. The association of T. gondii exposure with a history of surgery warrants for further research. Risk factors for T. gondii infection found in the present survey may help to design optimal educational programs to avoid T. gondii infection. PMID:26185685

Prevalence of Toxoplasma gondii antibodies in domestic donkeys (Equus asinus) in Durango, Mexico slaughtered for human consumption.

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Alvarado-Esquivel, Cosme; Alvarado-Esquivel, Domingo; Dubey, Jitender P

2015-01-17

Nothing is known about Toxoplasma gondii prevalence in donkeys in Mexico. Meat from donkey is consumed by humans in Mexico and also exported to other countries. We sought to determine the presence of antibodies against T. gondii in 239 domestic donkeys (Equus asinus) for slaughter in Durango, Mexico using the modified agglutination test (MAT). Donkeys were sampled in four premises (trade centers) where donkeys were gather for shipment to abattoirs in other Mexican states. Antibodies to T. gondii were found in 26 (10.9%) of 239 donkeys, with MAT titers of 1:25 in 7, 1:50 in 11, 1:100 in 6, and 1:200 in 2. Seropositive donkeys were found in three (75%) of the four gathering premises studied. Seroprevalence in donkeys varied from 0% to 23.1% among gathering premises. The seroprevalence of T. gondii infection was comparable among donkeys regardless their age, sex or health status. Seropositivity to T. gondii was found in donkeys between 1 to 12 years old. Multivariate analysis showed that seropositivity to T. gondii was associated with the gathering premises (OR = 1.58; 95% CI: 1.11-2.24; P = 0.009). This is the first report of T. gondii infection in donkeys in Mexico. Results indicate that consumption of undercooked or raw meat from T. gondii-infected donkeys is potentially a source of T. gondii infection for humans.

Seroprevalensi Toxoplasma gondii pada Kambing dan Bioassay Patogenitasnya pada Kucing

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Ni Made Yunik Novita Dewi Dewi

2013-11-01

Full Text Available Normal 0 false false false EN-US X-NONE X-NONE The study aimed to determine seroprevalence of Toxoplasmosis in goats sloughtered at Kampung Jawa, Denpasar, Bali and to evaluate their pathogenicities through bioassay in cats.One hundred serums and meats of goats were collected. Anti-Toxoplasma gondii antibody was determined using Indirect Haemaglutination (IHA test. The pathogenicity bioassay of Toxoplasma gondii was carried out through inoculating the meats of goats which had seropositive of Toxoplasma gondii to the cats. The pathogenicity was evaluated using the intensity of oocyte sheding from the cats. The result showed that the seroprevalence of Toxoplasmosis was 46%. There was not significant difference between pathogenicity of Toxoplasma gondii in cat inoculated with meat of goat which had a high and low titer of antibody against Toxoplasma gondii. /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; text-align:justify; line-height:150%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;}

Toxoplasma gondii: infection natural congenital in cattle and an experimental inoculation of gestating cows with oocysts.

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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.

Reciprocal moderation by Toxoplasma gondii seropositivity and blood phenylalanine - tyrosine ratio of their associations with trait aggression.

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Mathai, Ashwin Jacob; Lowry, Christopher A; Cook, Thomas B; Brenner, Lisa A; Brundin, Lena; Groer, Maureen W; Peng, Xiaoqing; Giegling, Ina; Hartmann, Annette M; Konte, Bettina; Friedl, Marion; Fuchs, Dietmar; Rujescu, Dan; Postolache, Teodor T

2016-12-01

We previously reported that trait aggression, proposed as an endophenotype for suicidal behavior, is positively associated with Toxoplasma gondii ( T. gondii ) seropositivity in females, but not in males. Additionally, older males seropositive for T. gondii had lower scores on measures of trait aggression, including self-aggression. Trait aggression may be influenced by dopaminergic signaling, which is known to be moderated by gender and age, and potentially enhanced in T. gondii positives through the intrinsic production of dopamine by the microorganism. Therefore, we investigated associations between trait aggression and interactions between T. gondii enzyme-linked immunoabsorbant assay (ELISA) IgG titer-determined seropositivity and high-performance liquid chromatography- (HPLC-) measured blood levels of dopamine precursors phenylalanine (Phe), tyrosine (Tyr), and their ratio in a sample of 1000 psychiatrically healthy participants. Aggressive traits were assessed using the questionnaire for measuring factors of aggression (FAF), the German version of the Buss-Durkee hostility questionnaire. We found that 1) the decrease in trait aggression scores in T. gondii -positive older males was only present in individuals with a low Phe:Tyr ratio, and 2) that there was a positive correlation between Phe:Tyr ratio and total aggression and selected subscales of aggression in T. gondii -positive males, but not in T. gondii -negative males. These findings point toward a gender-specific reciprocal moderation by Phe:Tyr ratio and T. gondii seropositivity of their associations with aggression scores, and lead to experimental interventions geared to manipulating levels of dopamine precursors in selected T. gondii positive individuals with increased propensity for aggression.

Detection and genotyping of Toxoplasma gondii DNA in the blood and milk of naturally infected donkeys (Equus asinus).

Science.gov (United States)

Mancianti, Francesca; Nardoni, Simona; Papini, Roberto; Mugnaini, Linda; Martini, Mina; Altomonte, Iolanda; Salari, Federica; D'Ascenzi, Carlo; Dubey, Jitender P

2014-04-03

Toxoplasma gondii is a worldwide zoonotic protozoan. Consumption of raw milk from infected animals is considered a risk factor for acquiring toxoplasmosis in humans. Recently, donkey milk has been indicated for therapeutic and nutritional purposes and T. gondii infection is common in donkeys. The purpose of the present paper was to detect the presence of parasite DNA in milk of T. gondii positive donkeys. Antibodies to T. gondii were found in 11 out of 44 healthy lactating donkeys by IFAT. T. gondii DNA was detected by PCR in blood of 6 and milk of 3 seropositive jennies. Results of limited RFLP-PCR genotyping indicated the presence of T. gondii genotype II or III, commonly found in Europe. The occurrence of T. gondii DNA in milk suggests that the consumption of raw milk from seropositive donkeys could be a potential source of human infection.

Mouse-virulent Toxoplasma gondii isolated from feral cats on Mona Island, Puerto Rico.

Science.gov (United States)

Dubey, J P; López-Torres, H Y; Sundar, N; Velmurugan, G V; Ajzenberg, D; Kwok, O C H; Hill, R; Dardé, M L; Su, C

2007-12-01

Cats are essential in the life cycle of Toxoplasma gondii because they are the only hosts that can excrete the environmentally resistant oocysts. Samples of serum, feces, and tissues from cats from Mona, a remote island off the coast of Puerto Rico, were examined for T. gondii infection. Antibodies to T. gondii were assayed by the modified agglutination test and found in 16 of 19 (84.2%) of cats, with titers of 1:10 in 2, 1:80 in 1, 1:160 in 4, 1:320 in 3, and 1:1,280 or higher in 6. Tissues of 19 of the 20 cats were bioassayed in mice for T. gondii infection. Toxoplasma gondii was isolated from tissues of 12 cats: from the hearts of 9, skeletal muscle of 10, and brain of 1 cat. All infected mice from 10 of 12 isolates died of acute toxoplasmosis during primary infection. Genotyping of these 12 T. gondii isolates (designated (TgCatPr 1-12) by 10 multilocus PCR-RFLP markers, i.e., SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and an apicoplast marker Apico, and the 6 multilocus microsatellite markers TUB2, W35, TgM-A, B18, B17, and M33, revealed 7 genotypes; 5 isolates had Type I alleles at all loci except at 1 microsatellite locus, and the remainder were atypical. The latter isolates of T. gondii were different biologically and phenotypically from the feline isolates from the rest of the Americas. One isolate (TgCatPr 12) was a mixed infection with 2 genotypes.

Seroepidemiology of Toxoplasma gondii in dogs in the State of Rio de Janeiro

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Nathalie Costa da Cunha

2016-11-01

Full Text Available ABSTRACT. Cunha N.C., Cordeiro M.D., Bravo S.A.C., Matos P.C.M., Almosny N.R.P. & da Fonseca A.H. [Seroepidemiology of Toxoplasma gondii in dogs in the State of Rio de Janeiro.] Soroepidemiologia de Toxoplasma gondii em cães no estado do Rio de Janeiro. Revista Brasileira de Medicina Veterinária 38(supl. 3: 114-121, 2016. Departamento de Saúde Coletiva Veterinária e Saúde Pública, Faculdade de Veterinária. Universidade Federal Fluminense, Vital Brazil, Niterói, RJ, Brazil. E-mail: nathaliecunha@id.uff.br Toxoplasmosis is a serious public health problem worldwide as it can cause prenatal and neonatal morbidity and mortality in humans. Although dogs are not definitive hosts of T. gondii, they play an important role in the mechanical dissemination of oocysts. This study aimed to carry out a seroepidemiological investigation of anti-Toxoplasma gondii antibodies in domestic dogs from seven municipalities in the state of Rio de Janeiro, Brazil. A crosssectional epidemiological study was carried out to evaluate the profile of anti-Toxoplasma gondii IgG antibodies in dogs from canine sera from different municipalities in the state of Rio de Janeiro. The municipalities studied were Cachoeiras de Macacu, Guapimirim, Itaboraí, Magé, Resende, Seropédica and Silva Jardim. The detection of antibodies of the IgG class anti-Toxoplasma gondii was performed using the indirect enzyme immunoadsorption (ELISA assay and the statistical analyzes used were the chi-square test and the prevalence ratio. Of the 651 samples tested, 300 were reactive for T. gondii, representing a relative frequency of 46.08% of seroreactive dogs. It was concluded that dogs are good sentinels for evaluations of risk for occurrence of T. gondii, emphasizing those coming from rural areas and that there was no difference in the occurrence of serorreative dogs in front of different municipalities studies of the state of Rio de Janeiro.

Seroprevalence of Toxoplasma gondii in wild boars, red deer and roe deer in Poland

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Witkowski Lucjan

2015-01-01

Full Text Available Little is known about the prevalence of Toxoplasma gondii in wild life, particularly game animals in Poland. Meat juice collected during the 2009/2010 and 2010/2011 hunting seasons from 552 red deer (Cervus elaphus, 367 wild boars (Sus scrofa and 92 roe deer (Capreolus capreolus was tested for T. gondii antibodies using the multi-species ID Screen Toxoplasmosis Indirect kit (IDvet, Montpellier, France. Antibodies to T. gondii were detected in 24.1% of red deer (95% CI: 20.7%, 27.8%, 37.6% of wild boar (95% CI: 32.8%, 42.7% and 30.4% of roe deer (95% CI: 22.0%, 40.5%. To the authors’ best knowledge, this is the first epidemiological report of T. gondii prevalence in red deer, roe deer and wild boars in Poland. T. gondii is present in wildlife animal tissues and consumption of the game may be a potential source of infection for humans.

Toxoplasma gondii exposes phosphatidylserine inducing a TGF-β1 autocrine effect orchestrating macrophage evasion

International Nuclear Information System (INIS)

Seabra, Sergio H.; Souza, Wanderley de; Matta, Renato A. da

2004-01-01

Toxoplasmosis is a worldwide disease caused by Toxoplasma gondii. Activated macrophages control T. gondii growth by nitric oxide (NO) production. However, T. gondii active invasion inhibits NO production, allowing parasite persistence. Here we show that the mechanism used by T. gondii to inhibit NO production persisting in activated macrophages depends on phosphatidylserine (PS) exposure. Masking PS with annexin-V on parasites or activated macrophages abolished NO production inhibition and parasite persistence. NO production inhibition depended on a transforming growth factor-β 1 (TGF-β 1 ) autocrine effect confirmed by the expression of Smad 2 and 3 in infected macrophages. TGF-β 1 led to inducible nitric oxide synthase (iNOS) degradation, actin filament (F-actin) depolymerization, and lack of nuclear factor-κB (NF-κB) in the nucleus. All these features were reverted by TGF-β 1 neutralizing antibody treatment. Thus, T. gondii mimics the evasion mechanism used by Leishmania amazonensis and also the anti-inflammatory response evoked by apoptotic cells

Epidemiology and pathology of Toxoplasma gondii in free-ranging California sea lions (Zalophus californianus)

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Carlson-Bremer, D; Colegrove, KM; Gulland, FMD; Conrad, PA; Mazet, JAK; Johnson, CK

2015-01-01

© Wildlife Disease Association 2015. The coccidian parasite Toxoplasma gondii infects humans and warm-blooded animals worldwide. The ecology of this parasite in marine systems is poorly understood, although many marine mammals are infected and susceptible to clinical toxoplasmosis. We summarized the lesions associated with T. gondii infection in the California sea lion (Zalophus californianus) population and investigated the prevalence of and risk factors associated with T. gondii exposure, a...

Epidemiology and pathology of Toxoplasma gondii in free-ranging California sea lions (Zalophus californianus).

Science.gov (United States)

Carlson-Bremer, Daphne; Colegrove, Kathleen M; Gulland, Frances M D; Conrad, Patricia A; Mazet, Jonna A K; Johnson, Christine K

2015-04-01

The coccidian parasite Toxoplasma gondii infects humans and warm-blooded animals worldwide. The ecology of this parasite in marine systems is poorly understood, although many marine mammals are infected and susceptible to clinical toxoplasmosis. We summarized the lesions associated with T. gondii infection in the California sea lion (Zalophus californianus) population and investigated the prevalence of and risk factors associated with T. gondii exposure, as indicated by antibody. Five confirmed and four suspected cases of T. gondii infection were identified by analysis of 1,152 medical records of necropsied sea lions from 1975-2009. One suspected and two confirmed cases were identified in aborted fetuses from a sea lion rookery. Toxoplasmosis was the primary cause of death in five cases, including the two fetuses. Gross and histopathologic findings in T. gondii-infected sea lions were similar to those reported in other marine mammals. The most common lesions were encephalitis, meningitis, and myocarditis. The antibody prevalence in stranded, free-ranging sea lions for 1998-2009 was 2.5% (±0.03%; IgG titer 640). There was an increase in odds of exposure in sea lions with increasing age, suggesting cumulative risk of exposure and persistent antibody over time. The occurrence of disseminated T. gondii infection in aborted fetuses confirms vertical transmission in sea lions, and the increasing odds of exposure with age is consistent with additional opportunities for horizontal transmission in free-ranging sea lions over time. These data suggest that T. gondii may have two modes of transmission in the sea lion population. Overall, clinical disease was uncommon in our study which, along with low prevalence of T. gondii antibody, suggests substantially less-frequent exposure and lower susceptibility to clinical disease in California sea lions as compared to sympatric southern sea otters (Enhydra lutris nereis).

Seroprevalence of Toxoplasma gondii from free-ranging black bears ( Ursus americanus ) from Florida.

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Chambers, D L; Ulrey, W A; Guthrie, J M; Kwok, O C H; Cox, J J; Maehr, D S; Dubey, J P

2012-06-01

Toxoplasma gondii is a significant worldwide parasitic protozoan. In the present study, prevalence of antibodies of T. gondii was examined from 29 free-ranging black bears ( Ursus americanus ) from south-central Florida where the host species was listed as state threatened during this project. Overall T. gondii prevalence was found to be 44.8%, specifically 46.2% in male and 43.8% in female U. americanus , using a modified agglutination test (1:25 titer). Seroprevalence differences between sexes were not significant (P > 0.05). Results of the present study add supportive data to the growing body of evidence suggesting that U. americanus has one of the highest T. gondii seroprevalences among all known intermediate hosts. In addition, our data emphasize the importance of understanding parasitic disease dynamics from a conservation perspective.

Molecular cloning and functional characterization of a rainbow trout liver Oatp

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Steiner, Konstanze, E-mail: konstanze.steiner@uni-konstanz.de [University of Konstanz, Human- and Environmental Toxicology, 78464 Konstanz (Germany); Hagenbuch, Bruno, E-mail: bhagenbuch@kumc.edu [Pharmacology, Toxicology and Therapeutics, The University of Kansas Medical Center, Kansas City 66160, KS (United States); Dietrich, Daniel R., E-mail: daniel.dietrich@uni-konstanz.de [University of Konstanz, Human- and Environmental Toxicology, 78464 Konstanz (Germany)

2014-11-01

Cyanobacterial blooms have an impact on the aquatic ecosystem due to the production of toxins (e.g. microcystins, MCs), which constrain fish health or even cause fish death. However the toxicokinetics of the most abundant toxin, microcystin-LR (MC-LR), are not yet fully understood. To investigate the uptake mechanism, the novel Oatp1d1 in rainbow trout (rtOatp1d1) was cloned, identified and characterized. The cDNA isolated from a clone library consisted of 2772 bp containing a 2115 bp open reading frame coding for a 705 aa protein with an approximate molecular mass of 80 kDa. This fish specific transporter belongs to the OATP1 family and has most likely evolved from a common ancestor of OATP1C1. Real time PCR analysis showed that rtOatp1d1 is predominantly expressed in the liver, followed by the brain while expression in other organs was not detectable. Transient transfection in HEK293 cells was used for further characterization. Like its human homologues OATP1A1, OATP1B1 and OATP1B3, rtOatp1d1 displayed multi-specific transport including endogenous and xenobiotic substrates. Kinetic analyses revealed a K{sub m} value of 13.9 μM and 13.4 μM for estrone-3-sulfate and methotrexate, respectively and a rather low affinity for taurocholate with a K{sub m} value of 103 μM. Furthermore, it was confirmed that rtOatp1d1 is a MC-LR transporter and therefore most likely plays a key role in the susceptibility of rainbow trout to MC intoxications. - Highlights: • A new Oatp1d1 in rainbow trout (rtOatp1d1) was cloned, identified and characterized. • rtOatp1d1 is predominantly expressed in the liver. • rtOatp1d1 displays multi-specific transport of endogenous and xenobiotic substrates. • rtOatp1d1 is a homologue of the OATP1A1, OATP1B1 and OATP1B3. • rtOatp1d1 is a microcystin (MC) transporter.

Molecular cloning and functional characterization of a rainbow trout liver Oatp

International Nuclear Information System (INIS)

Steiner, Konstanze; Hagenbuch, Bruno; Dietrich, Daniel R.

2014-01-01

Cyanobacterial blooms have an impact on the aquatic ecosystem due to the production of toxins (e.g. microcystins, MCs), which constrain fish health or even cause fish death. However the toxicokinetics of the most abundant toxin, microcystin-LR (MC-LR), are not yet fully understood. To investigate the uptake mechanism, the novel Oatp1d1 in rainbow trout (rtOatp1d1) was cloned, identified and characterized. The cDNA isolated from a clone library consisted of 2772 bp containing a 2115 bp open reading frame coding for a 705 aa protein with an approximate molecular mass of 80 kDa. This fish specific transporter belongs to the OATP1 family and has most likely evolved from a common ancestor of OATP1C1. Real time PCR analysis showed that rtOatp1d1 is predominantly expressed in the liver, followed by the brain while expression in other organs was not detectable. Transient transfection in HEK293 cells was used for further characterization. Like its human homologues OATP1A1, OATP1B1 and OATP1B3, rtOatp1d1 displayed multi-specific transport including endogenous and xenobiotic substrates. Kinetic analyses revealed a K m value of 13.9 μM and 13.4 μM for estrone-3-sulfate and methotrexate, respectively and a rather low affinity for taurocholate with a K m value of 103 μM. Furthermore, it was confirmed that rtOatp1d1 is a MC-LR transporter and therefore most likely plays a key role in the susceptibility of rainbow trout to MC intoxications. - Highlights: • A new Oatp1d1 in rainbow trout (rtOatp1d1) was cloned, identified and characterized. • rtOatp1d1 is predominantly expressed in the liver. • rtOatp1d1 displays multi-specific transport of endogenous and xenobiotic substrates. • rtOatp1d1 is a homologue of the OATP1A1, OATP1B1 and OATP1B3. • rtOatp1d1 is a microcystin (MC) transporter

Molecular cloning and characterization of pathogenesis-related ...

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use

2011-12-21

Dec 21, 2011 ... Available online at http://www.academicjournals.org/AJB ... November, 2011. We described the cloning and characterization of pathogenesis-related protein 5 gene in maize, named .... in two inbred lines was calculated using the ↵Ct method. .... Of the characterized PRs currently known, PR-1, PR-2,. PR-3 ...

Detection of Toxoplasma gondii oocysts in soils in northwestern China using a new semi-nested PCR assay.

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Wang, Meng; Meng, Peng; Ye, Qiang; Pu, Yuan-Hua; Yang, Xiao-Yu; Luo, Jian-Xun; Zhang, Nian-Zhang; Zhang, De-Lin

2014-09-28

Toxoplasma gondii is a zoonotic pathogen that can infect a range of animals and humans. Ingestion of T. gondii oocysts in soil is a significant transmission route for humans and animals acquiring toxoplasmosis. In the present study, we developed a new semi-nested PCR method to determine T. gondii oocysts distribution in soils in northwestern China. The one tube semi-nested PCR assay was developed to detect the oocysts of T. gondii in soil, targeting the repetitive 529 bp fragment of T. gondii genomic DNA. Then a total of 268 soil samples, including 148 samples from Gansu Province and 120 samples from Qinghai Province, northwestern China, were examined by the semi-nested PCR method. One third of the positive samples were sequenced. The sensitivity of the semi-nested PCR assay was 10(2)  T. gondii oocysts in 5 g soil sample. Investigation of soil samples from northwestern China showed that 34 out of 268 soil samples (12.69%) were T. gondii positive. Sequences of the partial 529 bp fragments varied from 0-1.2% among the sequenced samples. The prevalence of T. gondii oocysts in soil from cities (24/163) was slightly higher than that in soils from pasturing areas (10/105) (P = 0.21). Among the different regions in cities, the prevalence of T. gondii oocysts in soils from parks was 14.15%, whereas that in soils from schools was 19.05%. The present study firstly reported the prevalence of T. gondii oocysts in soils in northwest China using a novel semi-nested PCR assay, which provided baseline data for the effective prevention and control of toxoplasmosis in this region.

Seroprevalence, Detection of DNA in Blood and Milk, and Genotyping of Toxoplasma gondii in a Goat Population in Italy

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Francesca Mancianti

2013-01-01

Full Text Available Toxoplasma gondii is the causative agent of a major zoonosis with cosmopolitan distribution and is known to be transmitted mainly by the ingestion of undercooked or raw animal products. Drinking unpasteurized goat’s milk is a risk factor associated with human toxoplasmosis. However, very little is known about the excretion of DNA in goat milk. Aim of the present study was to determine the seroprevalence of T. gondii infection using a modified agglutination test (MAT, to detect T. gondii DNA by nested-PCR (n-PCR in samples of blood and milk from seropositive goats, and to genotype DNA isolates using 11 molecular markers in 127 adult lactating goats from 6 farms in Italy. Positive MAT results were found in 60.6% of goats while 13% of blood and milk samples from seropositive goats were positive to n-PCR. A kappa coefficient of 1 indicated a perfect agreement between blood and milk n-PCR. Genetic characterization of isolates revealed the occurrence of genotype III (, genotype I (, and atypical genotypes with hints for genotype I (. Our results suggest that the risk of excretion of Toxoplasma tachyzoites might frequently occur in milk of seropositive goats testing positive to n-PCR on blood.

Toxoplasma gondii peptide ligands open the gate of the HLA class I binding groove

DEFF Research Database (Denmark)

McMurtrey, Curtis; Trolle, Thomas; Sansom, Tiffany

2016-01-01

HLA class I presentation of pathogen-derived peptide ligands is essential for CD8+ T cell recognition of Toxoplasma gondii infected cells. Currently, little data exist pertaining to peptides that are presented after T. gondii infection. Herein we purify HLA-A*02:01 complexes from T. gondii infect...

Serotyping of Toxoplasma gondii in Cats (Felis domesticus) Reveals Predominance of Type II Infections in Germany

Science.gov (United States)

Background: Cats are definitive hosts of Toxoplasma gondii and play an essential role in the epidemiology of this parasite. The study aims at clarifying whether cats are able to develop specific antibodies against different clonal types of T. gondii and to determine by serotyping the T. gondii clona...

Incidence of adult brain cancers is higher in countries where the protozoan parasite Toxoplasma gondii is common

Science.gov (United States)

Thomas, Frédéric; Lafferty, Kevin D.; Brodeur, Jacques; Elguero, Eric; Gauthier-Clerc, Michel; Missé, Dorothée

2012-01-01

We explored associations between the common protozoan parasite Toxoplasma gondii and brain cancers in human populations. We predicted that T. gondii could increase the risk of brain cancer because it is a long-lived parasite that encysts in the brain, where it provokes inflammation and inhibits apoptosis. We used a medical geography approach based on the national incidence of brain cancers and seroprevalence of T. gondii. We corrected reports of incidence for national gross domestic product because wealth probably increases the ability to detect cancer. We also included gender, cell phone use and latitude as variables in our initial models. Prevalence of T. gondii explained 19 per cent of the residual variance in brain cancer incidence after controlling for the positive effects of gross domestic product and latitude among nations. Infection with T. gondii was associated with a 1.8-fold increase in the risk of brain cancers across the range of T. gondii prevalence in our dataset (4–67%). These results, though correlational, suggest that T. gondii should be investigated further as a possible oncogenic pathogen of humans.

[A review of the genomic and gene cloning studies in trees].

Science.gov (United States)

Yin, Tong-Ming

2010-07-01

Supported by the Department of Energy (DOE) of U.S., the first tree genome, black cottonwood (Populus trichocarpa), has been completely sequenced and publicly release. This is the milestone that indicates the beginning of post-genome era for forest trees. Identification and cloning genes underlying important traits are one of the main tasks for the post-genome-era tree genomic studies. Recently, great achievements have been made in cloning genes coordinating important domestication traits in some crops, such as rice, tomato, maize and so on. Molecular breeding has been applied in the practical breeding programs for many crops. By contrast, molecular studies in trees are lagging behind. Trees possess some characteristics that make them as difficult organisms for studying on locating and cloning of genes. With the advances in techniques, given also the fast growth of tree genomic resources, great achievements are desirable in cloning unknown genes from trees, which will facilitate tree improvement programs by means of molecular breeding. In this paper, the author reviewed the progress in tree genomic and gene cloning studies, and prospected the future achievements in order to provide a useful reference for researchers working in this area.

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.

Molecular cloning and protein structure of a human blood group Rh polypeptide

International Nuclear Information System (INIS)

Cherif-Zahar, B.; Bloy, C.; Le Van Kim, C.; Blanchard, D.; Bailly, P.; Hermand, P.; Salmon, C.; Cartron, J.P.; Colin, Y.

1990-01-01

cDNA clones encoding a human blood group Rh polypeptide were isolated from a human bone marrow cDNA library by using a polymerase chain reaction-amplified DNA fragment encoding the known common N-terminal region of the Rh proteins. The entire primary structure of the Rh polypeptide has been deduced from the nucleotide sequence of a 1384-base-pair-long cDNA clone. Translation of the open reading frame indicates that the Rh protein is composed of 417 amino acids, including the initiator methionine, which is removed in the mature protein, lacks a cleavable N-terminal sequence, and has no consensus site for potential N-glycosylation. The predicted molecular mass of the protein is 45,500, while that estimated for the Rh protein analyzed in NaDodSO 4 /polyacrylamide gels is in the range of 30,000-32,000. These findings suggest either that the hydrophobic Rh protein behaves abnormally on NaDodSO 4 gels or that the Rh mRNA may encode a precursor protein, which is further matured by a proteolytic cleavage of the C-terminal region of the polypeptide. Hydropathy analysis and secondary structure predictions suggest the presence of 13 membrane-spanning domains, indicating that the Rh polypeptide is highly hydrophobic and deeply buried within the phospholipid bilayer. These results suggest that the expression of the Rh gene(s) might be restricted to tissues or cell lines expressing erythroid characters

Role of NETs in the difference in host susceptibility to Toxoplasma gondii between sheep and cattle.

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Yildiz, Kader; Gokpinar, Sami; Gazyagci, Aycan Nuriye; Babur, Cahit; Sursal, Neslihan; Azkur, Ahmet Kursat

2017-07-01

The main aim of this study was to compare extracellular traps (NETs) formation by polymorphonuclear neutrophils (PMNs) of cattle and sheep when exposed to T. gondii tachyzoites in vitro. The effects of parasite concentrations and different incubation periods on NETs development in cattle and sheep PMNs were studied. The effect of NET structures on host cell invasion by tachyzoites was also studied. This is the first report of NETs development by sheep and cattle PMNs against T. gondii in vitro. T. gondii-induced extracellular DNA production from PMNs was dependent on tachyzoite concentrations and incubation time in both sheep and cattle. Many nuclear and cytoplasmic changes were observed in sheep and cattle PMNs after exposure to T. gondii tachyzoites. The typical appearance of NETs, with MPO, NE and histone (H3) attached to extracellular DNA, was observed. Tachyzoites were entrapped within this structure. Myeloperoxidase (MPO) activity was higher in the cattle PMN-tachyzoite co-cultures than sheep. NETs structures released from sheep PMNs caused mechanical immobilisation of T. gondii tachyzoites, however, NET structures released from cattle PMNs may be lethal to tachyzoites. Bovine MPO may have a lethal effect on T. gondii tachyzoites in vitro during a 3h incubation. Besides other mechanisms that effect on host susceptibility to T. gondii in sheep and cattle, extracellular traps formation as a part of immunological reactions may be play a role in host susceptibility to T. gondii. Copyright © 2017 Elsevier B.V. All rights reserved.

Influence of Toxoplasma Gondii Infection on Symptoms and Signs of Premenstrual Syndrome: A Cross-sectional Study

Science.gov (United States)

Alvarado-Esquivel, Cosme; Sánchez-Anguiano, Luis Francisco; Hernández-Tinoco, Jesús; Pérez-Álamos, Alma Rosa; Rico-Almochantaf, Yazmin del Rosario; Estrada-Martínez, Sergio; Vaquera-Enriquez, Raquel; Díaz-Herrera, Arturo; Ramos-Nevarez, Agar; Sandoval-Carrillo, Ada Agustina; Salas-Pacheco, José Manuel; Cerrillo-Soto, Sandra Margarita; Antuna-Salcido, Elizabeth Irasema; Liesenfeld, Oliver; Guido-Arreola, Carlos Alberto

2016-01-01

Infection with Toxoplasma gondii in brain may cause some symptoms that resemble those in women with premenstrual syndrome. To determine the association of T. gondii infection with symptoms and signs of premenstrual syndrome, we examined 489 women aged 30–40 years old. Sera of participants were analyzed for the presence of anti-Toxoplasma IgG and IgM antibodies using enzyme-linked immunoassays (EIA) and T. gondii DNA by polymerase chain reaction (PCR). Anti-T. gondii IgG antibodies were found in 38 (7.8%) of the women studied. Anti-T. gondii IgM antibodies were found in 13 (34.2%) of the 38 IgG seropositive women. Logistic regression showed two variables associated with seropositivity to T. gondii: presence of diarrhea (odds ratio [OR] = 6.10; 95% confidence interval [CI]: 1.37–27.85; P = 0.01) and weight gain (OR = 2.89; 95% CI: 1.37–6.07; P = 0.005), and two variables associated with high (>150 IU/ml) levels of IgG against T. gondii: presence of diarrhea (OR = 7.40; 95% CI: 1.79–30.46; P = 0.006) and abdominal inflammation (OR = 3.38; 95% CI: 1.13–10.10; P = 0.02). Positivity to EIA IgG and PCR was positively associated with obesity and negatively associated with joint pain by bivariate analysis. Our study for the first time reveals a potential association of T. gondii infection with clinical manifestations of premenstrual syndrome. PMID:27980858

A qualitative assessment of Toxoplasma gondii risk in ready-to-eat smallgoods processing.

Science.gov (United States)

Mie, Tanya; Pointon, Andrew M; Hamilton, David R; Kiermeier, Andreas

2008-07-01

Toxoplasma gondii is one of the most common parasitic infections of humans and other warm-blooded animals. In most adults, it does not cause serious illness, but severe disease may result from infection in fetuses and immunocompromised people. Consumption of raw or undercooked meats has consistently been identified as an important source of exposure to T. gondii. Several studies indicate the potential failure to inactivate T. gondii in the processes of cured meat products, This article presents a qualitative risk-based assessment of the processing of ready-to-eat smallgoods, which include cooked or uncooked fermented meat, pâté, dried meat, slow cured meat, luncheon meat, and cooked muscle meat including ham and roast beef. The raw meat ingredients are rated with respect to their likelihood of containing T. gondii cysts and an adjustment is made based on whether all the meat from a particular source is frozen. Next, the effectiveness of common processing steps to inactivate T. gondii cysts is assessed, including addition of spices, nitrates, nitrites and salt, use of fermentation, smoking and heat treatment, and the time and temperature during maturation. It is concluded that processing steps that may be effective in the inactivation of T. gondii cysts include freezing, heat treatment, and cooking, and the interaction between salt concentration, maturation time, and temperature. The assessment is illustrated using a Microsoft Excel-based software tool that was developed to facilitate the easy assessment of four hypothetical smallgoods products.

Seroprevalence of Toxoplasma gondii and Neospora caninum in red deer from Central Italy

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Guido Rocchigiani

2016-09-01

Full Text Available Neospora caninum and Toxoplasma gondii are cosmopolite protozoan parasites impacting on human and animal health. In particular, T. gondii commonly infects human beings and all warm-blooded animals, while N. caninum is responsible for bovine abortion and neuromuscular disease in dogs. The aim of the presented survey was to evaluate the occurrence and prevalence of these parasites in the most numerous Italian red deer population. The sera of 60 red deer ( Cervus elaphus inhabiting Central Italy (43°56’N 10°55’E and killed by selective hunting were examined using an indirect fluorescent antibody test (IFAT for both N. caninum and T. gondii antibodies. White blood cells (buffy coat were also checked by PCR and T. gondii DNA was genotyped. Thirteen out of 60 sera (22% scored positive for Toxoplasma, 17 samples (28% were Neospora positive. Coinfection was recorded in 5 cases (8%. T. gondii (genotype II and N. caninum DNA was detected in one and 3 samples of buffy coat, respectively. The presented study is the first to examine the occurrence of these parasites in the most numerous red deer Italian population, confirming this animal species as carrier of the investigated pathogens. These animals spread near human settlements, co-inhabiting with final hosts of [i]T. gondii[/i] and N. caninum and could contribute to their transmission to domestic ruminants and humans. In particular, the seroprevalence value for N. caninum was the highest among European records.

Toxoplasma gondii transmission by artificial insemination in sheep with experimentally contaminated frozen semen.

Science.gov (United States)

Consalter, Angélica; Silva, Andressa F; Frazão-Teixeira, Edwards; Matos, Luis F; de Oliveira, Francisco C R; Leite, Juliana S; Silva, Franciele B F; Ferreira, Ana M R

2017-03-01

Toxoplasma gondii is a parasite considered one of the major causes of reproductive problems in sheep. Furthermore, the presence of the agent in ram semen urges the possibility of sexual transmission in this species. The aim of this study was to evaluate if ram's frozen semen spiked with T. gondii tachyzoites would be able to cause infection in sheep by laparoscopic artificial insemination (AI). Nine ewes tested seronegative to anti-T. gondii antibodies by the modified agglutination test (MAT) were superovulated and inseminated to collect embryos. Animals were divided into two groups: G1 (n = 5), ewes inseminated with semen containing 4 × 10 7 tachyzoites; and G2 (n = 4), ewes inseminated with tachyzoite-free semen (control group). To confirm infection, ewe's blood samples were collected on days -14, -7, 0, 7, 14, 21, 28, 35, 49 and 57 after AI for analysis by MAT and PCR. Tissue samples of these ewes were also collected for histopathology and immunohistochemistry (IHC). Seven days after AI, all ewes of group G1 had specific antibodies to T. gondii, while those of G2 were negative. Toxoplasma gondii DNA was detected in the blood of one ewe and parasites were observed in tissues of all five animals inseminated with contaminated semen, indicating that semen freezing protocol does not affect T. gondii transmission by artificial insemination in sheep. Copyright © 2016 Elsevier Inc. All rights reserved.

The common parasite Toxoplasma gondii induces prostatic inflammation and microglandular hyperplasia in a mouse model.

Science.gov (United States)

Colinot, Darrelle L; Garbuz, Tamila; Bosland, Maarten C; Wang, Liang; Rice, Susan E; Sullivan, William J; Arrizabalaga, Gustavo; Jerde, Travis J

2017-07-01

Inflammation is the most prevalent and widespread histological finding in the human prostate, and associates with the development and progression of benign prostatic hyperplasia and prostate cancer. Several factors have been hypothesized to cause inflammation, yet the role each may play in the etiology of prostatic inflammation remains unclear. This study examined the possibility that the common protozoan parasite Toxoplasma gondii induces prostatic inflammation and reactive hyperplasia in a mouse model. Male mice were infected systemically with T. gondii parasites and prostatic inflammation was scored based on severity and focality of infiltrating leukocytes and epithelial hyperplasia. We characterized inflammatory cells with flow cytometry and the resulting epithelial proliferation with bromodeoxyuridine (BrdU) incorporation. We found that T. gondii infects the mouse prostate within the first 14 days of infection and can establish parasite cysts that persist for at least 60 days. T. gondii infection induces a substantial and chronic inflammatory reaction in the mouse prostate characterized by monocytic and lymphocytic inflammatory infiltrate. T. gondii-induced inflammation results in reactive hyperplasia, involving basal and luminal epithelial proliferation, and the exhibition of proliferative inflammatory microglandular hyperplasia in inflamed mouse prostates. This study identifies the common parasite T. gondii as a new trigger of prostatic inflammation, which we used to develop a novel mouse model of prostatic inflammation. This is the first report that T. gondii chronically encysts and induces chronic inflammation within the prostate of any species. Furthermore, T. gondii-induced prostatic inflammation persists and progresses without genetic manipulation in mice, offering a powerful new mouse model for the study of chronic prostatic inflammation and microglandular hyperplasia. © 2017 Wiley Periodicals, Inc.

Seroprevalence and Associated Risk Factors for Toxoplasma gondii Infection in Healthy Blood Donors: A Cross-Sectional Study in Sonora, Mexico.

Science.gov (United States)

Alvarado-Esquivel, Cosme; Rascón-Careaga, Antonio; Hernández-Tinoco, Jesús; Corella-Madueño, María Alba Guadalupe; Sánchez-Anguiano, Luis Francisco; Aldana-Madrid, María Lourdes; Velasquez-Vega, Edgar; Quizán-Plata, Trinidad; Navarro-Henze, José Luis; Badell-Luzardo, Joel Alberto; Gastélum-Cano, José María; Liesenfeld, Oliver

2016-01-01

Toxoplasma gondii (T. gondii) can be transmitted by blood transfusion. We determined the prevalence of T. gondii infection in healthy blood donors in Hermosillo city, Mexico, and the association of infection with T. gondii with the sociodemographic, clinical, and behavioral characteristics of blood donors. Four hundred and eight blood donors who attended two public blood banks in Hermosillo city were examined for anti-T. gondii IgG and IgM antibodies by using enzyme-linked immunoassays. Of the 408 blood donors (mean age 31.77 ± 9.52; range 18-60 years old) studied, 55 (13.5%) were positive for anti-T. gondii IgG antibodies, and 12 (21.8%) of them were also positive for anti-T. gondii IgM antibodies. Multivariate analysis showed that seropositivity to T. gondii was associated with age (OR = 1.74; 95% CI: 1.03-2.94; P = 0.03) and tobacco use (OR = 2.09; 95% CI: 1.02-4.29; P = 0.04). Seropositivity to T. gondii was correlated with the number of pregnancies, deliveries, and cesarean sections. The seroprevalence of T. gondii infection in blood donors in Sonora is the highest reported in blood donors in northern Mexico so far. This is the first report of an association of T. gondii exposure and tobacco use. Further research to confirm this association is needed.

Seroprevalence of Toxoplasma gondii and Neospora caninum infections in goats from two Argentinean provinces

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María Laura Gos

2017-11-01

Full Text Available The aim of this study was to compare the seroprevalence of Toxoplasma gondii and Neospora caninum in goats from two Argentinean provinces raised under different management conditions. A total of 2922 serum samples from adult goats of Córdoba (n=2187 and Buenos Aires provinces (n= 735, Argentina, were assayed by indirect fluorescence antibody test (IFAT, cut-off 1:100 for antibodies to T. gondii and N. caninum. Seroprevalence was 40.8% (CI 39.0%-42.6% and 5.5% (CI 4.7%-6.4% for T. gondii and N. caninum, respectively. The seropositivity for both infections was higher in goats from dairy farms, resulting as follows: for T. gondii 32.7% (CI 30.8%-34.8% in extensive farms and 59.3% (CI 56.1%-62.6% in dairy farms and for N. caninum 4.1% (CI 3.2%-4.9% in extensive farms and 8.8% (CI 6.9%-10.7% in dairy farms. This is the first extensive seroepidemiology investigation for T. gondii and N. caninum in goats in Argentina.

Long-Term Protective Immune Response Elicited by Vaccination with an Expression Genomic Library of Toxoplasma gondii

OpenAIRE

Fachado, Alberto; Rodriguez, Alexandro; Molina, Judith; Silvério, Jaline C.; Marino, Ana P. M. P.; Pinto, Luzia M. O.; Angel, Sergio O.; Infante, Juan F.; Traub-Cseko, Yara; Amendoeira, Regina R.; Lannes-Vieira, Joseli

2003-01-01

Immunization of BALB/c mice with an expression genomic library of Toxoplasma gondii induces a Th1-type immune response, with recognition of several T. gondii proteins (21 to 117 kDa) and long-term protective immunity against a lethal challenge. These results support further investigations to achieve a multicomponent anti-T. gondii DNA vaccine.

Molecular cloning and characterization of genes required for nucleotide excision repair in yeast

International Nuclear Information System (INIS)

Friedberg, E.C.

1987-01-01

Nucleotide excision repair in the yeast S. cerevisiae is a complex process which involves a large number of genes. At least five of these genes (RAD1, RAD2, RAD3, RAD4 and RAD10) are absolutely required for this process and mutations in any of these genes result in no detectable excision repair in vivo. In order to understand the function of these genes in DNA repair, the authors isolated a number of them by screening a yeast genomic library for recombinant plasmids which complement the phentoype of sensitivity to ultraviolet (UV) radiation imparted to mutant strains. A plasmid containing the RAD4 gene was isolated by an alternative strategy which will be discussed. The cloned genes have been extensively characterized. It has been determined that the RAD3 gene is essential for the viability of haploid yeast cells in the absence of DNA damage. The RAD2 gene is inducible by treatment of cells with a variety of DNA-damaging agents, including UV radiation and ionizing radiation. The RAD10 gene shares considerable amino acid sequence homology with a cloned gene involved in nucleotide excision repair in human cells. Yeast is a particularly versatile organism for studying gene function by molecular and genetic approaches and emphasis is placed on many of the techniques used in the present studies

Computational Prediction of MicroRNAs from Toxoplasma gondii Potentially Regulating the Hosts’ Gene Expression

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Müşerref Duygu Saçar

2014-10-01

Full Text Available MicroRNAs (miRNAs were discovered two decades ago, yet there is still a great need for further studies elucidating their genesis and targeting in different phyla. Since experimental discovery and validation of miRNAs is difficult, computational predictions are indispensable and today most computational approaches employ machine learning. Toxoplasma gondii, a parasite residing within the cells of its hosts like human, uses miRNAs for its post-transcriptional gene regulation. It may also regulate its hosts’ gene expression, which has been shown in brain cancer. Since previous studies have shown that overexpressed miRNAs within the host are causal for disease onset, we hypothesized that T. gondii could export miRNAs into its host cell. We computationally predicted all hairpins from the genome of T. gondii and used mouse and human models to filter possible candidates. These were then further compared to known miRNAs in human and rodents and their expression was examined for T. gondii grown in mouse and human hosts, respectively. We found that among the millions of potential hairpins in T. gondii, only a few thousand pass filtering using a human or mouse model and that even fewer of those are expressed. Since they are expressed and differentially expressed in rodents and human, we suggest that there is a chance that T. gondii may export miRNAs into its hosts for direct regulation.

Inactivation of Toxoplasma gondii on blueberries using low dose irradiation without affecting quality

Science.gov (United States)

Toxoplasma gondii is a common protozoan parasite, whose environmentally-resistant stage, the oocyst, can contaminate irrigation water and fresh edible produce. Current washing steps in produce processing may not be effective for eliminating T. gondii from at-risk varieties of produce. The objective ...

Prevalence of Toxoplasma gondii IgG and IgM and associated risk ...

African Journals Online (AJOL)

Prevalence of Toxoplasma gondii IgG and IgM and associated risk factors among HIV-positive and HIV-negative patients in Vhembe district of South Africa. ... shown a high prevalence of T. gondii (IgG) among patients attending different HIV clinics in the Vhembe district with no current infections among pregnant women.

Atypical Toxoplasma gondii genotype in feral cats from the Fernando de Noronha Island, northeastern Brazil.

Science.gov (United States)

Melo, R P B; Almeida, J C; Lima, D C V; Pedrosa, C M; Magalhães, F J R; Alcântara, A M; Barros, L D; Vieira, R F C; Garcia, J L; Mota, R A

2016-07-15

Toxoplasma gondii isolates from Brazil have a different phenotypic and genotypic pattern, with predominance of virulent isolates and recombinant genotypes, compared to the North Hemisphere. Considering that a new T. gondii genotype, non-pathogenic to mice, was previously identified from free-range chickens from the Fernando de Noronha Island, Brazil, this study aimed to identify genotypes of this parasite in tissue samples of feral cats (Felis catus) from this Brazilian Island. Anti-T. gondii IgG antibodies were detected in 18/31 (58%) feral cats. Two non-virulent T. gondii isolates were obtained by mouse bioassay. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1, L358 and Apico) and an atypical strain of T. gondii (ToxoDB #146) was identified. This is the first report of this genotype in feral cats. Copyright © 2016 Elsevier B.V. All rights reserved.

Infection rate of toxoplasma gondii and age distribution in female patients with sterility

International Nuclear Information System (INIS)

Li Shuhong; Dai Pei; Cui Liming; Zong Shan; Zuo Wenjing

2006-01-01

Objective: To discuss the relationship between the infection of Toxoplasma gondii and female sterility. Methods: Toxoplasma gondii serum antibody were determined in 882 women with sterility (experimental group) and 107 normal bearing women (control group) by using ELISA. At the same time the differences of the infection with Toxoplasma gondii between the ages of the sterility women were analyzed. Results: The positive rate in experimental group was 15.87% (140/882), the positive rate in control group was 5.61% (6/107), remarkable difference was found between two groups (P<0.01). The infection rate in the different age groups (20-24, 25-29, 30-34, 35-39 and ≥40) is 5.63%, 15.24%, 17.91%, 19.44% and 15.38%. Conclusion: Toxoplasma gondii infection may be one of the factors which can cause sterility, and the infection rates at different ages have no instinct differences. (authors)

Seropositivity and risk factors associated with Toxoplasma gondii infection in wild birds from Spain.

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Oscar Cabezón

Full Text Available Toxoplasma gondii is a zoonotic intracellular protozoan parasite of worldwide distribution that infects many species of warm-blooded animals, including birds. To date, there is scant information about the seropositivity of T. gondii and the risk factors associated with T. gondii infection in wild bird populations. In the present study, T. gondii infection was evaluated on sera obtained from 1079 wild birds belonging to 56 species (including Falconiformes (n=610, Strigiformes (n=260, Ciconiiformes (n=156, Gruiformes (n=21, and other orders (n=32, from different areas of Spain. Antibodies to T. gondii (modified agglutination test, MAT titer ≥1:25 were found in 282 (26.1%, IC(95%:23.5-28.7 of the 1079 birds. This study constitute the first extensive survey in wild birds species in Spain and reports for the first time T. gondii antibodies in the griffon vulture (Gyps fulvus, short-toed snake-eagle (Circaetus gallicus, Bonelli's eagle (Aquila fasciata, golden eagle (Aquila chrysaetos, bearded vulture (Gypaetus barbatus, osprey (Pandion haliaetus, Montagu's harrier (Circus pygargus, Western marsh-harrier (Circus aeruginosus, peregrine falcon (Falco peregrinus, long-eared owl (Asio otus, common scops owl (Otus scops, Eurasian spoonbill (Platalea leucorodia, white stork (Ciconia ciconia, grey heron (Ardea cinerea, common moorhen (Gallinula chloropus; in the International Union for Conservation of Nature (IUCN "vulnerable" Spanish imperial eagle (Aquila adalberti, lesser kestrel (Falco naumanni and great bustard (Otis tarda; and in the IUCN "near threatened" red kite (Milvus milvus. The highest seropositivity by species was observed in the Eurasian eagle owl (Bubo bubo (68.1%, 98 of 144. The main risk factors associated with T. gondii seropositivity in wild birds were age and diet, with the highest exposure in older animals and in carnivorous wild birds. The results showed that T. gondii infection is widespread and can be at a high level in many wild

Simultaneous detection of viruses and Toxoplasma gondii in cerebrospinal fluid specimens by multiplex polymerase chain reaction-based reverse hybridization assay.

Science.gov (United States)

Del Prete, Raffaele; Di Taranto, Anna Maria; Lipsi, Maria Rosaria; Natalicchio, Maria Iole; Antonetti, Raffaele; Miragliotta, Giuseppe

2009-04-01

The lack of rapidity and the low sensitivity and specificity of traditional laboratory methods limits their usefulness in the laboratory diagnosis of viral central nervous system (CNS) infections. This study describes the use of a commercially available multiplex polymerase chain reaction (mPCR)-based reverse hybridization assay (RHA) for the simultaneous detection of the genomes of 8 viruses and Toxoplasma gondii in cerebrospinal fluids (CSF) from 181 patients suspected of having viral meningitis. Twenty-two/181 (12.15%) CSF samples resulted positive by mPCR. Eighteen/22 were positive for 1 viral pathogen, whereas a dual infection was detected in 4/22 samples. Epstein-Barr virus (EBV) was the most commonly detected virus (6/22), followed by herpes simplex virus type-1 (HSV-1) (5/22) and -2 (HSV-2) (4/22). Cytomegalovirus (CMV), human herpesvirus-6 (HHV-6), and Epstein-Barr virus (EBV) were detected in 1 specimen each. Two CSF samples were co-infected by HSV-1/HSV-2, 1 sample by HHV-6/T. gondii, and 1 sample by EBV/EV, respectively. Our data support the usefulness of mPCR as a rapid molecular method for the simultaneous detection of major viral pathogens and T. gondii in aseptic meningitis also to allow the earlier application of specific antiviral therapy.

Molecular cloning and characterization of the plasma membrane ...

African Journals Online (AJOL)

O. violaceus) was cloned. The full-length cDNA of O. violaceus gene (OvPIP) was 1314 bp and contained 1188 open reading frame encoding a protein of 395 amino acids. Homology analysis revealed that OvPIP strongly resembled other PIP ...

The transcriptome of Toxoplasma gondii

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Roos David S

2005-12-01

Full Text Available Abstract Background Toxoplasma gondii gives rise to toxoplasmosis, among the most prevalent parasitic diseases of animals and man. Transformation of the tachzyoite stage into the latent bradyzoite-cyst form underlies chronic disease and leads to a lifetime risk of recrudescence in individuals whose immune system becomes compromised. Given the importance of tissue cyst formation, there has been intensive focus on the development of methods to study bradyzoite differentiation, although the molecular basis for the developmental switch is still largely unknown. Results We have used serial analysis of gene expression (SAGE to define the Toxoplasma gondii transcriptome of the intermediate-host life cycle that leads to the formation of the bradyzoite/tissue cyst. A broad view of gene expression is provided by >4-fold coverage from nine distinct libraries (~300,000 SAGE tags representing key developmental transitions in primary parasite populations and in laboratory strains representing the three canonical genotypes. SAGE tags, and their corresponding mRNAs, were analyzed with respect to abundance, uniqueness, and antisense/sense polarity and chromosome distribution and developmental specificity. Conclusion This study demonstrates that phenotypic transitions during parasite development were marked by unique stage-specific mRNAs that accounted for 18% of the total SAGE tags and varied from 1–5% of the tags in each developmental stage. We have also found that Toxoplasma mRNA pools have a unique parasite-specific composition with 1 in 5 transcripts encoding Apicomplexa-specific genes functioning in parasite invasion and transmission. Developmentally co-regulated genes were dispersed across all Toxoplasma chromosomes, as were tags representing each abundance class, and a variety of biochemical pathways indicating that trans-acting mechanisms likely control gene expression in this parasite. We observed distinct similarities in the specificity and

Anti-Toxoplasma gondii secretory IgA in tears of patients with ocular toxoplasmosis: immunodiagnostic validation by ELISA

OpenAIRE

Lynch,Maria Isabel; Malagueño,Elizabeth; Lynch,Luiz Felipe; Ferreira,Silvana; Stheling,Raphael; Oréfice,Fernando

2009-01-01

Toxoplasma gondii causes posterior uveitis and the specific diagnosis is based on clinical criteria. The presence of anti-T. gondii secretory IgA (sIgA) antibodies in patients' tears has been reported and an association was found between ocular toxoplasmosis and the anti-T. gondii sIgA isotype in Brazilian patients. The purpose of this study was to provide an objective validation of the published ELISA test for determining the presence of anti-T. gondii sIgA in the tears of individuals with o...

Sequence Variation in Toxoplasma gondii rop17 Gene among Strains from Different Hosts and Geographical Locations

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Nian-Zhang Zhang

2014-01-01

Full Text Available Genetic diversity of T. gondii is a concern of many studies, due to the biological and epidemiological diversity of this parasite. The present study examined sequence variation in rhoptry protein 17 (ROP17 gene among T. gondii isolates from different hosts and geographical regions. The rop17 gene was amplified and sequenced from 10 T. gondii strains, and phylogenetic relationship among these T. gondii strains was reconstructed using maximum parsimony (MP, neighbor-joining (NJ, and maximum likelihood (ML analyses. The partial rop17 gene sequences were 1375 bp in length and A+T contents varied from 49.45% to 50.11% among all examined T. gondii strains. Sequence analysis identified 33 variable nucleotide positions (2.1%, 16 of which were identified as transitions. Phylogeny reconstruction based on rop17 gene data revealed two major clusters which could readily distinguish Type I and Type II strains. Analyses of sequence variations in nucleotides and amino acids among these strains revealed high ratio of nonsynonymous to synonymous polymorphisms (>1, indicating that rop17 shows signs of positive selection. This study demonstrated the existence of slightly high sequence variability in the rop17 gene sequences among T. gondii strains from different hosts and geographical regions, suggesting that rop17 gene may represent a new genetic marker for population genetic studies of T. gondii isolates.

First report of Toxoplasma gondii infection in market-sold adult chickens, ducks and pigeons in northwest China

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Cong Wei

2012-06-01

Full Text Available Abstract Background Toxoplasma gondii infection is a global concern, affecting a wide range of warm-blooded animals and humans worldwide, including poultry. Domestic and companion birds are considered to play an important role in the transmission of T. gondii to humans and other animals. However, little information on T. gondii infection in domestic birds in Lanzhou, northwest China was available. Therefore, this study was performed to determine the seroprevalence of T. gondii infection in domestic birds in Lanzhou, northwest China. Methods In the present study, the seroprevalence of T. gondii antibodies in 413 (305 caged and 108 free-range adult chickens, 334 (111 caged and 223 free-range adult ducks and 312 adult pigeons in Lanzhou, northwest China, were examined using the modified agglutination test (MAT. Results 30 (7.26% chickens, 38 (11.38% ducks and 37 (11.86% pigeons were found to be positive for T. gondii antibodies at the cut-off of 1:5. The prevalences in caged and free-range chickens were 6.23% and 10.19% respectively, however, statistical analysis showed that the difference was not significant (P > 0.05. The seroprevalences in caged and free-range ducks were 6.31% and 13.90% respectively, but the difference was not statistically significant (P > 0.05. Conclusions The results of the present survey indicated the presence of T. gondii infection in adult chickens, ducks and pigeons sold for meat in poultry markets in Lanzhou, northwest China, which poses a potential risk for T. gondii infection in humans and other animals in this region. This is the first seroprevalence study of T. gondii infection in domestic birds in this region.

Molecular Cloning And Sequencing Of Disintegrin Like Domain ...

African Journals Online (AJOL)

Disintegrin-like domain was cloned and sequenced from Cerastes cerastes venom gland tissue. Nested RT-PCR was performed using initial primers designed based on the homology of disintegrins from Trimeresurus flavoviridis, Glodius halys , Agkistrodon halys and Trimeresurus macrosquamatus. The homology was ...

Construction and characterisation of a full-length infectious molecular clone from a fast replicating, X4-tropic HIV-1 CRF02.AG primary isolate

International Nuclear Information System (INIS)

Tebit, Denis M.; Zekeng, Leopold; Kaptue, Lazare; Kraeusslich, Hans-Georg; Herchenroeder, Ottmar

2003-01-01

Based on our previous analysis of HIV-1 isolates from Cameroon, we constructed a full-length infectious molecular clone from a primary isolate belonging to the CRF02.AG group of recombinant viruses which dominate the HIV-epidemic in West and Central Africa. The virus derived by transfection of the proviral clone pBD6-15 replicated with similar efficiency compared to its parental isolate and used CXCR4 as coreceptor as well. Furthermore, HIV-1 BD6-15 exhibited similar replication properties and virus yield as the reference B-type HIV-1 strain NL4-3. Sequence analysis revealed open reading frames for all structural and accessory genes apart from vpr. Phylogenetic and bootscanning analyses confirmed that BD6-15 clusters with CRF02.AG recombinant strains from West and Central Africa with similar cross-over points as described for the CRF02.AG prototype strain lbNG. Thus, pBD6-15 represents the first non-subtype B infectious molecular clone of a fast replicating, high producer, X4-tropic primary HIV-1 isolate, which had only been briefly passaged in primary cells

A quantitative microbial risk assessment for meatborne Toxoplasma gondii infection in The Netherlands

NARCIS (Netherlands)

Opsteegh, M.; Prickaerts, S.; Frankena, K.; Evers, E.G.

2011-01-01

Toxoplasma gondii is an important foodborne pathogen, and the cause of a high disease burden due to congenital toxoplasmosis in The Netherlands. The aim of this study was to quantify the relative contribution of sheep, beef and pork products to human T. gondii infections by Quantitative Microbial

Seroprevalence and isolation of viable Toxoplasma gondii from raptors in the southeastern USA

Science.gov (United States)

Raptors are good indicators of the prevalence of Toxoplasma gondii in the environment because they prey on small mammals and birds. These prey species are a major source of infection in felids, which shed the environmentally resistant oocysts. We assessed T. gondii infection in 281 opportunistically...

A meta-analysis of the prevalence of toxoplasma gondii igg antibodies in patients with mental disorders

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Angela Dragomir1,

2016-12-01

Full Text Available OBJECTIVES AND BACKGROUND Toxoplasma gondii infection has been recently associated with schizophrenia and other psychiatric disorders. The aim of the present study was to evaluate the prevalence of T. gondii infection in psychiatric patients by using meta-analytical methods. MATERIALS AND METHODS By systematic research of PUBMED Database, we identified several articles on this issue. We included casecontrol studies focused on the seroprevalence of T. gondii (IgG antibodies in patients with psychiatric disorders and healthy controls, published over the past 10 years R3.2.2. free software for statistical computing and graphics was used to perform the meta-analysis. Data were pooled using a random effects model and Mantel-Haenszel method. RESULTS The PUBMED Database showed references to only seven scientific papers that investigated the prevalence of T. gondii IgG antibodies in psychiatric patients. Six hundred seventy three patients and seven hundred seventy four controls coped with the inclusion criteria and were used in our analysis. We found a significant increase of T. gondii IgG antibodies in patients with schizophrenia and other psychiatric disorders compared with controls (41.6% vs 24.54%, OR = 2.16, 95% CI = [1.45-3.21], P = .001. CONCLUSIONS An increased seroprevalence of T. gondii IgG antibodies has been reported in psychiatric patients. Our study suggests that T. gondii infection may be relevant in order to determine and understand the complex etiology of schizophrenia and other psychiatric disorders. Graphical abstract: Meta-analysis of the prevalence of T. gondii IgG antibodies in patients with mental disorders REFERENCES 1. Hamidinejat H, Ghorbanpoor M, Hosseini H, Alavi SM, Nabavi L, Jalali MH, Borojeni MP, Jafari H, Mohammadaligol S. Toxoplasma gondii infection in first-episode and inpatient individuals with schizophrenia. Int J Infect Dis. 2010;14:e978-81. doi: 10.1016/j.ijid.2010.05.018. 2. Alvarado-Esquivel C, Carrillo-Oropeza D

Canine distemper virus and Toxoplasma gondii co-infection in dogs with neurological signs

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D.M. Aguiar

2012-02-01

Full Text Available O presente estudo relata a ocorrência de co-infecção entre o vírus da cinomose canina (CDV e Toxoplama gondii em cães com sinais neurológicos. Amostras de soro e tecido nervoso (pos-mortem de 21 cães, suspeitos de cinomose canina foram analisadas pela Reação de Imunofluorecência indireta (RIFI para pesquisa de anticorpos contra T. gondii e N. caninum e por RT-PCR para CDV. Dezessete (80,9% cães foram positivos para o CDV pela RT-PCR e 8 (38,1% foram positivos para anticorpos contra T. gondii. Sete cães (41,1% apresentaram-se positivos para ambos agentes, caracterizando processo de co-infecção. Somente 1 (4,7% cão foi soropositivo para N. caninum (RIFI=100, entretanto este mesmo animal foi positivo para T. gondii (RIFI=4096 e para CDV (RT-PCR.

Seroepidemiology of Toxoplasma gondii Infection in Women of Reproductive Age: A Cross-Sectional Study in a Northwestern Mexican City

Science.gov (United States)

Alvarado-Esquivel, Cosme; Corella-Madueno, Maria Alba Guadalupe; Hernandez-Tinoco, Jesus; Rascon-Careaga, Antonio; Sanchez-Anguiano, Luis Francisco; Martinez-Robinson, Karla Guadalupe; Aldana-Madrid, Maria Lourdes; Quizan-Plata, Trinidad; Canez-Carrasco, Maria Guadalupe; Perez-Martinez, Cinthia Jhovanna

2018-01-01

Background Through a cross-sectional survey, we determined the seroprevalence and correlates of Toxoplasma gondii (T. gondii) infection in women of reproductive age in Hermosillo City, Mexico. Methods We studied 445 women of reproductive age in Hermosillo City in the northwestern Mexican state of Sonora. Women were enrolled in the University of Sonora. Sera of women were examined for IgG and IgM antibodies to T. gondii by commercially available enzyme immunoassays. The association of T. gondii seropositivity with the characteristics of the pregnant women was determined by bivariate and multivariate analyses. Results Of the 445 women (mean age: 22.18 ± 5.6 years) studied, 16 (3.6%) had IgG antibodies to T. gondii, and two (12.5%) were also positive for IgM antibodies to T. gondii. Of the 16 anti-T. gondii IgG-positive women, six (37.5%) had IgG levels higher than 150 IU/mL, four (25.0%) between 100 and 150 IU/mL, and six (37.5%) between 9 and 99 IU/mL. Multivariate analysis of socio-demographic and behavioral variables showed that T. gondii seropositivity was associated with older age (odds ratio (OR): 5.30; 95% confidence interval (CI): 1.37 - 20.50; P = 0.01) and boar meat consumption (OR: 6.86; 95% CI: 1.27 - 37.07; P = 0.02). Conclusions Women of reproductive age in Hermosillo City had a low seroprevalence of T. gondii infection. However, this finding indicates that most of these women were susceptible to a primary infection. Factors associated with T. gondii infection found in this study may be useful for the optimal planning of preventive measures against T. gondii infection and its sequelae. PMID:29416579

Prevalence of Toxoplasma gondii in Chicken samples from delta of Egypt using ELISA, histopathology and immunohistochemistry.

Science.gov (United States)

Ibrahim, Hany M; Abdel-Ghaffar, Fathy; Osman, Gamalat Y; El-Shourbagy, Safinaz H; Nishikawa, Yoshifumi; Khattab, Reham A

2016-06-01

Estimates of the zoonotic diseases are helpful for monitoring and improving public health. Laboratory-based surveillance provides crucial information for assessing zoonotic disease trends and developments. Toxoplasmosis is considered as a zoonotic disease and has both medical and veterinary importance since it leads to abortion in humans and several animal species. In view of the worldwide importance of T. gondii, this study aimed to estimate the prevalence of T. gondii in chickens from the Delta of Egypt. A total of 304 blood and brain samples were collected from Egyptian chickens from Gharbiya, Qalyoubiya, Minufiya, Beheira, Kafr EL-Shaykh and Dakahlia Provinces. In order to determine the serological and histopathological prevalence of T. gondii, the samples were examined by ELISA, histopathology and immunohistochemistry (IHC). The prevalence of T. gondii was 11.18, 6.91, 6.91 % by ELISA, histopathology and IHC, respectively. Statistically significant differences in the prevalence of T. gondii were observed on the basis of season, sex and habitat. These data provide valuable information regarding the epidemiology of T. gondii infections in Egyptian chickens, which can be employed in developing efficient strategies for disease management and control.

Toxoplasma gondii and Epilepsy.

Science.gov (United States)

Ayaz, Erol; Türkoğlu, Şule Aydın; Orallar, Hayriye

2016-06-01

Toxoplasma gondii is a zoonotic parasite can be seen in all the vital organ; in the acute phase, it can be found in the blood, cerebrospinal fluid, semen, tears, saliva, urine, and in almost all body fluids. Transplasental infection can lead to fetal damage and miscarriage. Its last hosts are felines and intermediate hosts are all mammals, including humans. People infected by the ingestion of meat containing cysts in undercooked or raw, are thrown oocysts with cat felines By taking in water and food, from mother to fetus transplacental way, the infected organ transplantation, blood transfusion, laboratory accidents and kaprofaj transmitted by mechanical vectors of the invertebrates. Suppression of the immune system is being transformed to the shape and texture of the cysts with bradyzoite. The parasite settles in the cells of the tissue cysts and causes change in the cellular mechanisms, such as cytokinin task. Depending on changes and type of neurotransmitter (GABA, glutamate, serotonin, dopamine) levels in CSF in ions (Ca, K, Cl, Mg), it is believed that there is a change in their concentration. In this review, literature about the relationship between T. gondii and epilepsy and epileptiform activity the importance of parasites, which settle in the brain, will be highlighted.

New, previously unreported correlations between latent Toxoplasma gondii infection and excessive ethanol consumption.

Science.gov (United States)

Samojłowicz, Dorota; Borowska-Solonynko, Aleksandra; Kruczyk, Marcin

2017-11-01

A number of world literature reports indicate that a latent Toxoplasma gondii infection leads to development of central nervous system disorders, which in turn may lead to altered behavior in the affected individuals. T. gondii infection has been observed to play the greatest role in drivers, suicides, and psychiatric patients. Studies conducted for this manuscript involve a different, never before really reported correlation between latent T. gondii infection and ethanol abuse. A total of 538 decedents with a known cause of death were included in the study. These individuals were divided into three groups: the risky behavior group, inconclusively risky behavior group, and control group. The criterion for this division was the likely effect of the individual's behavior on the mechanism and cause of his/her death. The material used for analyses were blood samples collected during routine medico-legal examinations in these cases. The blood samples were used to measure anti-T. gondii IgG antibodies with an enzyme-linked immunosorbent assay (ELISA). Moreover, the following data were recorded for each decedent: sex, age, circumstances of death, cause of death, time from death to autopsy, and (if provided) substance abuse status (alcohol, illicit drugs). In those cases where blood alcohol level or toxicology tests were requested by the Prosecutor's Office, their results were also included in our analysis. Test results demonstrated a strong correlation between latent T. gondii infection and engaging in risky behaviors leading to death. Moreover, analyses demonstrated a positive correlation between the presence of anti-T. gondii IgG antibodies and psychoactive substance (especially ethanol) abuse, however, the causal relationship remains unclear. Due to the fact that alcohol abuse constitutes a significant social problem, searching for eliminable risk factors for addiction is extremely important. Our analyses provided new important information on the possible effects of

Molecular cloning, expression and characterization of Pru a 1, the major cherry allergen.

Science.gov (United States)

Scheurer, S; Metzner, K; Haustein, D; Vieths, S

1997-06-01

A high percentage of birch pollen allergic patients experiences food hypersensitivity reactions after ingestion of several fruits and vegetables. Previous work demonstrated common epitopes on an allergen of Mr 18,000 from sweet cherry (Prunus avium) and Bet v 1, the major allergen from birch pollen. N-terminal amino acid sequencing showed a sequence identity of 67% with Bet v 1. Here we report the cloning and cDNA sequencing of this cherry allergen. The entire deduced amino acid sequence described a protein of Mr 17,700 with 59.1% identity to Bet v 1. High degrees of identity in the range of 40 to 60% were also found with related allergens from other kinds of tree pollen and plant foods as well as with stress-induced proteins from food plants such as parsley, potato and soya. The coding DNA of the cherry protein was cloned into vector pET-16b and expressed in E. coli strain BL21(DE3) as a His-tag fusion protein. As shown by SDS-PAGE, the apparent molecular masses of the nonfusion protein and the natural allergen were identical. The fusion protein showed high IgE binding potency when sera from patients allergic to cherry were tested by immunoblotting and enzyme allergosorbent tests. Moreover, it cross-reacted strongly with IgE specific for the natural counterpart and for Bet v 1. The high biological activity of the recombinant fusion protein was further confirmed by the induction of a strong histamine release in basophils from cherry-allergic patients. Since sera from 17/19 of such patients contained IgE against this allergen it was classified as a major allergen and named Pru a 1. Recombinant Pru a 1 mimics most of the allergenic potency of cherry extract and hence could be a useful tool for studying the molecular and immunological properties of pollen related food allergens.

Low Seroprevalence of Leishmania infantum and Toxoplasma gondii in the Horse Population in Israel.

Science.gov (United States)

Aharonson-Raz, Karin; Baneth, Gad; Lopes, Ana Patrícia; Brancal, Hugo; Schallig, Henk; Cardoso, Luís; Steinman, Amir

2015-12-01

A cross-sectional investigation was done on the seroprevalence of Leishmania infantum and Toxoplasma gondii infection among apparently healthy horses in Israel. This survey included 383 horses distributed in 22 farms throughout Israel during the years 2011-2013. Serum samples were tested for the presence of immunoglobulin G (IgG) antibodies using the direct agglutination test (DAT) specific to Leishmania and by the modified agglutination test (MAT) for the presence of IgG antibodies to T. gondii. Low seroprevalences were detected for both L. infantum and T. gondii in the horse population in Israel; of the 338 horses tested, 6 (1.4%) were found to be seropositive for L. infantum and 11 (2.5%) for T. gondii, with no significant association between seroprevalence and demographic/environmental factors. An ongoing geographical expansion of L. infantum, previously reported in humans and dogs in Israel, was also supported by our results in horses. Here we present evidence of exposure of horses to L. infantum and T. gondii in Israel. Continuous seroprevalence surveillance in horses, such as the one performed in this study, might further elucidate the eco-epidemiology of these two important zoonotic parasites in this country.

MOLECULAR DETECTION AND CLONING FOR RICKETTSIA-LIKE BACTERIA OF MILKY HAEMOLYMPH DISEASE OF SPINY LOBSTER Panulirus spp.

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Isti Koesharyani

2017-01-01

Full Text Available Spiny lobster (Panulirus homarus and Panulirus ornatus are important commodities for Indonesia. The aquaculture of lobster is susceptible for several diseases like parasite, fungi, bacteria, and virus. Among those diseases, milky haemolymph disease (MHD is often seen as a symptom to mass mortality occurred at lobster farms in Gerupuk Bay of Lombok. The purpose of this study was to determine the lobster diseases on cage culture in Gerupuk Bay of Lombok, West Nusa Tenggara. The study was undertaken from January to March 2015. Diseases status was determined by application of molecular plat-form, polymerase chain reaction (PCR with designation of specific primer for MHD (254F/R, 254F: 5’-CGA-GGA-CCA-GAG-ATG-GAC-CTT-3’ and 254R: 5’-GCT-CAT-TGT-CAC-CGC-CAT-TGT-3’ with PCR size product of 254 bp. and for cloned the pathogen was used TA-cloning Invitrogen for the DNA plasmid as positive control for other analysis. Several tissue samples i.e hepatopancreas, haemolymph, part of muscle hepatopancreas P. homarus and P. ornatus were taken from cage culture farms at Gerupuk Bay then preserved on 90% ethanol for further analysis by PCR and then the amplificated DNA were cloned into pCR®2.1 plasmid and transformed into competent E. coli. The result showed that almost all lobster samples from Gerupuk Bay were positive infected by MHD, as the results of PCR amplification whereas the band appeared at 254bp. Also MHD plasmid has been successfully cloned and will be used for further examination. Histopathologically in hepatopancreas infection have seen necrosis that contain numerous of rickettsia-like bacteria.

Prevalence of Toxoplasma gondii antibodies and intestinal parasites in stray cats from Nigde, Turkey

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Bengi Dündar

2010-01-01

Full Text Available The prevalence of antibodies to Toxoplasma gondii was investigated by the Sabin-Feldman Dye test (SFDT in 72 stray cats from Nigde, Turkey. A total of 55 (76.4% of the analysed sera had antibodies to T. gondii. The seropositivity of T. gondii was 77.1% in male and 75.7% in female cats (P>0.05. Faeces of these cats were also examined by zinc sulphate flotation method for the presence of parasite oocysts and eggs of other parasites. Two protozoan parasites were identified as Isospora spp. (12.5% and Eimeria spp. (4.1% in cats. Toxoplasma gondii oocysts were not found in any faecal samples analysed. Two parasitic helminth species were observed: Toxocara cati (15.2% and Toxascaris leonina (20.8%. These common ascarids were recorded for the first time in cats from Nigde.

Serology and histopathology of Toxoplasma gondii and Neospora caninum in dogs with neurologic disorders/ Sorologia e histopatologia de Toxoplasma gondii e Neospora caninum em cães portadores de distúrbios neurológicos

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Thiago Neves Batista

2002-05-01

Full Text Available The evolvement of Toxoplasma gondii and Neospora caninum on etiology of clinical-neurological signs in dogs was investigated in this work. Ninety-eight urban dogs with neurologic disturbances were divided into two groups: Group I consisted of 67 dogs examined by the Animal Neurologic Service of Universidade Estadual de Londrina, between 1987 and 1990. Group II consisted of 31 dogs examined on the same University between 2000 and 2001. Dogs were serological evaluated for evidence of T. gondii or N. caninum. The serological results for T. gondii were 82,5% and 35.4% on groups I and II, respectively. Histologic lesions compatible with distemper on the two groups were observed. T. gondii cysts were observed in two dogs by histopathology and immunohistochemistry. Neosporosis was not observed in these dogs, possibly because their urban origin.Neste trabalho estudou-se a participação do Toxoplasma gondii e Neospora caninum na etiologia de sinais neurológicos em cães. Foram utilizados 98 cães com distúrbios neurológicos provenientes da área urbana divididos em dois grupos: Grupo I constituído de 67 animais atendidos no período de 1987 a 1990 pelo Serviço de Neurologia Animal da Universidade Estadual de Londrina, onde se utilizaram os tecidos emblocados em parafina e Grupo II com 31 animais atendidos no período de 2000 a 2001 nesta mesma instituição. Os animais foram avaliados sorologicamente para pesquisa de anticorpos contra T. gondii e N. caninum. Os resultados da sorologia para T. gondii obtidos no grupo I e II foram 82,5% e 35.4%, respectivamente. Foram observadas lesões histológicas compatíveis com cinomose nos dois grupos. Cistos de T. gondii foram diagnosticados em dois cães através da histopatologia e imuno-histoquímica. A neosporose não foi diagnosticada em nenhum animal, provavelmente devido à procedência urbana dos animais estudados.

Effect of 3-bromopyruvate and atovaquone on infection during in vitro interaction of Toxoplasma gondii and LLC-MK2 cells.

Science.gov (United States)

de Lima, Loyze Paola O; Seabra, Sergio H; Carneiro, Henrique; Barbosa, Helene S

2015-09-01

Toxoplasma gondii infection can be severe during pregnancy and in immunocompromised patients. Current therapies for toxoplasmosis are restricted to tachyzoites and have little or no effect on bradyzoites, which are maintained in tissue cysts. Consequently, new therapeutic alternatives have been proposed as the use of atovaquone has demonstrated partial efficacy against tachyzoites and bradyzoites. This work studies the effect of 3-bromopyruvate (3-BrPA), a compound that is being tested against cancer cells, on the infection of LLC-MK2 cells with T. gondii tachyzoites, RH strain. No effect of 3-BrPA on host cell proliferation or viability was observed, but it inhibited the proliferation of T. gondii. The incubation of cultures with lectin Dolichos biflorus agglutinin (DBA) showed the development of cystogenesis, and an ultrastructural analysis of parasite intracellular development confirmed morphological characteristics commonly found in tissue cysts. Moreover, the presence of degraded parasites and the influence of 3-BrPA on endodyogeny were observed. Infected cultures were alternatively treated with a combination of this compound plus atovaquone. This resulted in a 73% reduction in intracellular parasites after 24 h of treatment and a 71% reduction after 48 h; cyst wall formation did not occur in these cultures. Therefore, we conclude that the use of 3-BrPA may serve as an important tool for the study of (i) in vitro cystogenesis; (ii) parasite metabolism, requiring a deeper understanding of the target of action of this compound on T. gondii; (iii) the alternative parasite metabolic pathways; and (iv) the molecular/cellular mechanisms that trigger parasite death. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Toxoplasma gondii infections in sheep in Sicily, southern Italy

DEFF Research Database (Denmark)

Vesco, G; Buffolano, W; La Chiusa, S

2007-01-01

is very prevalent, and eating unprocessed sheep and lamb meat has a high risk of transmitting infections to humans. The presence of cats on the farm, farm size and using surface water as drinking water for the animals were risk factors for infection in sheep, with age as a significant confounder...... outdoor on pasture and only one was claiming organic farming. Having cats on the farm, age of the animals, farm size and the use of surface water sources for drinking were all significantly associated with T. gondii-infected animals on the farm. T. gondii infection in mutton used for human consumption...

Seroprevalence of Toxoplasma gondii in free-ranging and feral cats on Amami Oshima Island, Japan

OpenAIRE

MATSUU, Aya; YOKOTA, Shin-ichi; ITO, Keiko; MASATANI, Tatsunori

2017-01-01

On Amami Oshima Island, free-ranging and feral cats are harmful to wildlife populations. In this study, the seroprevalence of Toxoplasma gondii in these cats was examined using a newly developed Gaussia luciferase immunoprecipitation system assay. Of 1,363 cats, 123 cats (9.0%) was positive for T. gondii. The prevalence was significantly different in different areas; among cats in the rural area, where many wild animals live, including endangered species, T. gondii infection was more prevalen...

An Aspergillus oryzae acetyl xylan esterase: molecular cloning and characteristics of recombinant enzyme expressed in Pichia pastoris.

Science.gov (United States)

Koseki, Takuya; Miwa, Yozo; Akao, Takeshi; Akita, Osamu; Hashizume, Katsumi

2006-02-10

We screened 20,000 clones of an expressed sequence tag (EST) library from Aspergillus oryzae (http://www.nrib.go.jp/ken/EST/db/index.html) and obtained one cDNA clone encoding a protein with similarity to fungal acetyl xylan esterase. We also cloned the corresponding gene, designated as Aoaxe, from the genomic DNA. The deduced amino acid sequence consisted of a putative signal peptide of 31-amino acids and a mature protein of 276-amino acids. We engineered Aoaxe for heterologous expression in P. pastoris. Recombinant AoAXE (rAoAXE) was secreted by the aid of fused alpha-factor secretion signal peptide and accumulated as an active enzyme in the culture medium to a final level of 190 mg/l after 5 days. Purified rAoAXEA before and after treatment with endoglycosidase H migrated by SDS-PAGE with a molecular mass of 31 and 30 kDa, respectively. Purified rAoAXE displayed the greatest hydrolytic activity toward alpha-naphthylacetate (C2), lower activity toward alpha-naphthylpropionate (C3) and no detectable activity toward acyl-chain substrates containing four or more carbon atoms. The recombinant enzyme catalyzed the release of acetic acid from birchwood xylan. No activity was detectable using methyl esters of ferulic, caffeic or sinapic acids. rAoAXE was thermolabile in comparison to other AXEs from Aspergillus.

Investigating seagrass in Toxoplasma gondii transmission in Florida (Trichechus manatus latirostris) and Antillean (T. m. manatus) manatees

Science.gov (United States)

Wyrosdick, Heidi M; Gerhold, Richard; Su, Chunlei; Mignucci-Giannoni, Antonio A.; Bonde, Robert K.; Chapman, Alycia; Riviera-Perez, Carla; Martinez, Jessica; Miller, Debra L.

2017-01-01

Toxoplasma gondii is a feline protozoan reported to cause morbidity and mortality in manatees and other marine mammals. Given the herbivorous nature of manatees, ingestion of oocysts from contaminated water or seagrass is presumed to be their primary mode of infection. The objectives of this study were to investigate oocyst contamination of seagrass beds in Puerto Rico and determine the seroprevalence of T. gondii in Antillean (Trichechus manatus manatus) and Florida (T. m. latirostris) manatees. Sera or plasma from Antillean (n = 5) and Florida (n = 351) manatees were tested for T. gondii antibodies using the modified agglutination test. No T. gondii DNA was detected via PCR in seagrass samples (n = 33) collected from Puerto Rico. Seroprevalence was 0%, suggesting a lower prevalence of T. gondii in these manatee populations than previously reported. This was the first study to investigate the potential oocyst contamination of the manatee diet, and similar studies are important for understanding the epidemiology of T. gondii in herbivorous marine mammals.

Use of filter papers to determine seroprevalence of Toxoplasma gondii among hunted ungulates in remote Peruvian Amazon.

Science.gov (United States)

Aston, Emily J; Mayor, Pedro; Bowman, Dwight D; Mohammed, Hussni O; Liotta, Janice L; Kwok, Oliver; Dubey, J P

2014-04-01

Toxoplasmosis is a zoonosis caused by the protozoan Toxoplasma gondii, and it is found worldwide. To determine whether ungulates are reservoirs of T. gondii in an isolated and remote region of the northeastern Peruvian Amazon, antibodies to T. gondii were determined in 5 species of ungulates by the modified agglutination test (MAT). These animals were hunted by subsistence hunters along the Yavarí-Mirín River, in the northeastern Peruvian Amazon. Blood samples were collected by hunters on filter papers. For determination of T. gondii antibodies, blood was eluted from filter papers, and a titer of 1:25 was considered indicative of exposure to T. gondii. Antibodies to T. gondii were found in 26 (31.0%) peccaries (Pecari tajacu, Tayassu pecari), six (17.1%) brocket deer (Mazama americana, Mazama gouazoubira), and four (40.0%) lowland tapir (Tapirus terrestris). We also introduced a modification to the MAT protocol that allows the extraction of fluid samples from several types of laboratory-grade filter paper, thus enabling researchers to easily adapt their approaches to the materials presented to them.

Use of filter papers to determine seroprevalence of Toxoplasma gondii among hunted ungulates in remote Peruvian Amazon☆

Science.gov (United States)

Aston, Emily J.; Mayor, Pedro; Bowman, Dwight D.; Mohammed, Hussni O.; Liotta, Janice L.; Kwok, Oliver; Dubey, J.P.

2013-01-01

Toxoplasmosis is a zoonosis caused by the protozoan Toxoplasma gondii, and it is found worldwide. To determine whether ungulates are reservoirs of T. gondii in an isolated and remote region of the northeastern Peruvian Amazon, antibodies to T. gondii were determined in 5 species of ungulates by the modified agglutination test (MAT). These animals were hunted by subsistence hunters along the Yavarí-Mirín River, in the northeastern Peruvian Amazon. Blood samples were collected by hunters on filter papers. For determination of T. gondii antibodies, blood was eluted from filter papers, and a titer of 1:25 was considered indicative of exposure to T. gondii. Antibodies to T. gondii were found in 26 (31.0%) peccaries (Pecari tajacu, Tayassu pecari), six (17.1%) brocket deer (Mazama americana, Mazama gouazoubira), and four (40.0%) lowland tapir (Tapirus terrestris). We also introduced a modification to the MAT protocol that allows the extraction of fluid samples from several types of laboratory-grade filter paper, thus enabling researchers to easily adapt their approaches to the materials presented to them. PMID:24918073

21 CFR 866.3780 - Toxoplasma gondii serological reagents.

Science.gov (United States)

2010-04-01

... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3780 Toxoplasma... (immunofluorescent reagents) used to identify Toxoplasma gondii from clinical specimens. The identification aids in...

Usefulness of the detection of Toxoplasma gondii antigens in AIDS patients Utilidad de la detección de antígenos de Toxoplasma gondii en pacientes con SIDA

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Alberto Fachado

1994-12-01

Full Text Available Toxoplasmic encephalitis (TE is a mayor cause of central nervous system infection in patients with acquired immunodeficiency syndrome (AIDS. Toxoplasma antibodies were detected in 56 of 79 patients with AIDS (71%, in the present study. Fourteen out of 57 seropositive patients developed TF (25% and had Toxoplasma gondii antigen detected in their urine. For this, most of them received an effective therapy, with the subsequent disappearance of the symptoms and discontinuity of excretion of the T. gondii antigens. Our results suggest that the monitoring of T. gondii antigen in the urine of AIDS patients may be useful to decide on the proper time for therapy, as well as to avoid the beginning of neurologic signs in these patients.La Encefalitis Toxoplásmica (ET es la más importante complicación infecciosa del Sistema Nervioso Central en pacientes de SIDA. Anticuerpos anti-Toxoplasma gondii fueron detectados en 57 de 79 pacientes de SIDA (71%. De estos seropositivos, desarrollaron la enfermedad (ET 14 (25%, en los que coincidentemente se detectó la presencia de antígeno del parásito en orina y por tanto fueron objeto de una terapia efectiva, con la subsecuente desaparición de los sintomas y de los antígenos excretados. Por los resultados del presente trabajo, consideramos lo útil de monitorear en estos pacientes la presencia de antígenos de T. gondii con el objetivo de aplicar oportunamente métodos quimoprofilácticos que eviten el surgimiento de manifestaciones neurológicas en estos pacientes.

Production, characterization and applications for Toxoplasma gondii-specific polyclonal chicken egg yolk immunoglobulins.

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Álvaro Ferreira Júnior

Full Text Available Toxoplasma gondii may cause abortions, ocular and neurological disorders in warm-blood hosts. Immunized mammals are a wide source of hyperimmune sera used in different approaches, including diagnosis and the study of host-parasite interactions. Unfortunately, mammalian antibodies present limitations for its production, such as the necessity for animal bleeding, low yield, interference with rheumatoid factor, complement activation and affinity to Fc mammalian receptors. IgY antibodies avoid those limitations; therefore they could be an alternative to be applied in T. gondii model.In this study we immunized hens with soluble tachyzoite antigens of T. gondii (STAg and purified egg yolk antibodies (IgY by an inexpensive and simple method, with high yield and purity degree. IgY anti-STAg antibodies presented high avidity and were able to recognize a broad range of parasite antigens, although some marked differences were observed in reactivity profile between antibodies produced in immunized hens and mice. Interestingly, IgY antibodies against Neospora caninum and Eimeria spp. did not react to STAg. We also show that IgY antibodies were suitable to detect T. gondii forms in paraffin-embedded sections and culture cell monolayers.Due to its cost-effectiveness, high production yield and varied range of possible applications, polyclonal IgY antibodies are useful tools for studies involving T. gondii.

High prevalence of toxoplasmosis in cats from Egypt: isolation of viable Toxoplasma Gondii, tissue distribution, and isolate designation

Science.gov (United States)

Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that excrete environmentally resistant oocysts in feces. In the present study, 158 feral cats from Giza, Egypt were examined for T. gondii infection. Antibodies to T. gondii were found in 97.4% with modified ...

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

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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. gondii

Frequency of Toxoplasma gondii in the retina in eye banks in Brazil.

Science.gov (United States)

Costa, Deise F; Nascimento, Heloisa; Sutili, Aline; Nobrega, Fernando A J; Fowler, Flavio; Nobrega, Mario Junqueira; Garrido, Cristina; de Oliveira Dias, Janaina; Adán, Consuelo B D; Rizzo, Luiz Vicente; Silveira, Claudio; Belfort, Rubens; Commodaro, Alessandra G

2017-07-01

Ocular toxoplasmosis is the main cause of posterior uveitis worldwide frequently leading to vision loss. In Brazil, the seroprevalence of Toxoplasma gondii infection ranges from 50 to 80% depending of the region studied. The frequency of toxoplasmic retinal scar may reach 18% of the adults in the South of Brazil. Our goal was to determine the frequency of T. gondii DNA in retinas from eye banks from different regions in Brazil. A total of 162 eyes were obtained from eye banks in Manaus (n = 60), Sao Paulo (n = 60), Chapeco (n = 26), and Joinville (n = 16). The retinas were macroscopically analyzed and collected for DNA extraction. Real-time PCR (qPCR) was performed using the T. gondii B1 marker. By qPCR, a higher frequency of T. gondii DNA in the retinas from the eye bank of Joinville (25%) was found when compared to Manaus (5%). The retinas from Sao Paulo and Chapeco were qPCR negative. Clinical examination determined the retina lesions to be compatible with toxoplasmosis in the following frequencies: Joinville (62.5%), Manaus (10%), Sao Paulo (6.7%), and Chapeco (15.4%).

Lacrimal secretory IgA in active posterior uveitis induced by Toxoplasma gondii.

Science.gov (United States)

Lynch, Maria Isabel; Cordeiro, Francisco; Ferreira, Silvana; Ximenes, Ricardo; Oréfice, Fernando; Malagueño, Elizabeth

2004-12-01

It is quite difficult to diagnose active toxoplasmosis in patients with ocular toxoplasmosis. Active posterior uveitis presumably due to Toxoplasma gondii infection (APUPT) is seldom produced during a prime-infection; hence most patients do not show high IgM antibodies. High levels of IgA have been described in active toxoplasmosis. The purpose of this study was to investigate possible association between APUPT and the specific anti-parasite sIgA in tears. The study was carried out as case-control. Tears of 25 clinically confirmed APUPT patients and 50 healthy control subjects were analyzed. All were IgG seropositive. Specific sIgA was determined by ELISA assay using T. gondii RH strain crude extract. Anti-T. gondii sIgA was found in 84% of the cases and in 22% of the control subjects. The intensity of the reaction was higher in APUPT cases (P = 0.007). There was strong association between APUPT patients and lacrimal sIgA (odds-ratio 18.61, P = 0.0001). ELISA test sensitivity was 84% and specificity 78%. Our data suggest that anti-T.gondii secretory IgA found in tears may become an important marker for active ocular toxoplasmosis.

Aborto espontâneo e toxoplasmose ocular em um casal infectado com Toxoplasma gondii = Spontaneous abortion and ocular toxoplasmosis in a couple infected with Toxoplasma gondii

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Souza, Carla Zangari de

2015-01-01

Full Text Available Objetivos: Relatar os casos de um casal, em que a esposa apresentou toxoplasmose na gestação e o esposo, após 15 anos, foi diagnosticado com toxoplasmose ocular. Relato dos casos: Gestante, grávida de gêmeos, residente de um estado na região Sul do Brasil. No primeiro mês de gestação apresentou sorologia não reagente para IgG e IgM anti-Toxoplasma gondii. Na sétima semana de gestação houve soroconversão (IgM e IgG anti-T. gondii reagentes e aborto natural. O esposo, após 15 anos desde o episódio da esposa, queixou-se de visão embaçada/irritada. O exame de fundoscopia revelou lesões típicas de coriorretinite recidivante por toxoplasmose no olho direito. Os exames sorológicos foram IgG anti-T. gondii reagente e IgM anti-T. gondii não reagente. Conclusões: Este relato, de um casal com desfechos clínicos importantes de toxoplasmose, em diferentes formas, enfatiza a relevância do problema da toxoplasmose, que nas suas diversas manifestações, pode trazer graves consequências para a vida de uma família

Molecular cloning and characterization of strictosidine synthase, a ...

African Journals Online (AJOL)

Mitragynine is one of the most dominant indole alkaloids present in the leaves of Mitragyna speciosa, a species of Rubiaceae. This alkaloid is believed to be synthesized via condensation of the amino acid derivative, tryptamine and secologanine by the action of strictosidine synthase (STR). The cDNA clone encoding STR ...

Molecular genetic transfection of the coccidian parasite Sarcocystis neurona.

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Gaji, Rajshekhar Y; Zhang, Deqing; Breathnach, Cormac C; Vaishnava, Shipra; Striepen, Boris; Howe, Daniel K

2006-11-01

Sarcocystis neurona is an apicomplexan parasite that is the major cause of equine protozoal myeloencephalitis (EPM). The biology of this pathogen remains poorly understood in part due to unavailability of molecular genetic tools. Hence, with an objective to develop DNA transfection capabilities for S. neurona, the 5' flanking region of the SnSAG1 gene was isolated from a genomic library and used to construct expression plasmids. In transient assays, the reporter molecules beta-galactosidase (beta-gal) and yellow fluorescent protein (YFP) could be detected in electroporated S. neurona, thereby confirming the feasibility of transgene expression in this organism. Stable transformation of S. neurona was achieved using a mutant dihydrofolate reductase thymidylate synthase (DHFR-TS) gene of Toxoplasma gondii that confers resistance to pyrimethamine. This selection system was used to create transgenic S. neurona that stably express beta-gal and YFP. As shown in this study, these transgenic clones can be useful for analyzing growth rate of parasites in vitro and for assessing drug sensitivities. More importantly, the DNA transfection methods described herein should greatly facilitate studies examining intracellular parasitism by this important coccidian pathogen.

toxoMine: an integrated omics data warehouse for Toxoplasma gondii systems biology research.

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Rhee, David B; Croken, Matthew McKnight; Shieh, Kevin R; Sullivan, Julie; Micklem, Gos; Kim, Kami; Golden, Aaron

2015-01-01

Toxoplasma gondii (T. gondii) is an obligate intracellular parasite that must monitor for changes in the host environment and respond accordingly; however, it is still not fully known which genetic or epigenetic factors are involved in regulating virulence traits of T. gondii. There are on-going efforts to elucidate the mechanisms regulating the stage transition process via the application of high-throughput epigenomics, genomics and proteomics techniques. Given the range of experimental conditions and the typical yield from such high-throughput techniques, a new challenge arises: how to effectively collect, organize and disseminate the generated data for subsequent data analysis. Here, we describe toxoMine, which provides a powerful interface to support sophisticated integrative exploration of high-throughput experimental data and metadata, providing researchers with a more tractable means toward understanding how genetic and/or epigenetic factors play a coordinated role in determining pathogenicity of T. gondii. As a data warehouse, toxoMine allows integration of high-throughput data sets with public T. gondii data. toxoMine is also able to execute complex queries involving multiple data sets with straightforward user interaction. Furthermore, toxoMine allows users to define their own parameters during the search process that gives users near-limitless search and query capabilities. The interoperability feature also allows users to query and examine data available in other InterMine systems, which would effectively augment the search scope beyond what is available to toxoMine. toxoMine complements the major community database ToxoDB by providing a data warehouse that enables more extensive integrative studies for T. gondii. Given all these factors, we believe it will become an indispensable resource to the greater infectious disease research community. © The Author(s) 2015. Published by Oxford University Press.

Seroepidemiology of Toxoplasma gondii infection in bats from São Paulo city, Brazil

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Toxoplasmosis is a parasitic zoonosis caused by Toxoplasma gondii with a worldwide distribution. The aim of this study was to determine the frequency of IgG anti-T. gondii antibodies in bats from São Paulo city, Brazil. A total of 616 serum samples were collected from 22 species of bats. Anti-T. gon...

Transplacental transmission of Toxoplasma gondii in minipigs infected with strains of different virulence

DEFF Research Database (Denmark)

Jungersen, Gregers; Bille-Hansen, Vivi; Jensen, Lene Bai

2001-01-01

Infections with the Zoonotic protozoan Toxoplasma gondii during pregnancy can result in severe fetal infections. To investigate the use of pigs as animal models for congenital toxoplasmosis, tachyzoites of 5 T. gondii strains, with low to intermediate virulence in mice, were intravenously...... animal models fur studies of transplacental transmission and pathogenesis of congenital toxoplasmosis....

Lack of Association Between Toxoplasma gondii Infection and Diabetes Mellitus: A Matched Case-Control Study in a Mexican Population.

Science.gov (United States)

Alvarado-Esquivel, Cosme; Loera-Moncivais, Nayely; Hernandez-Tinoco, Jesus; Sanchez-Anguiano, Luis Francisco; Hernandez-Madrid, Guillermina; Rabago-Sanchez, Elizabeth; Centeno-Tinoco, Maria Magdalena; Sandoval-Carrillo, Ada A; Salas-Pacheco, Jose M; Campos-Moreno, Oscar Vladimir; Antuna-Salcido, Elizabeth Irasema

2017-06-01

Very little is known about the association between infection with Toxoplasma gondii ( T. gondii ) and diabetes mellitus. We perform an age- and gender-matched case-control study to determine the association of T. gondii infection and diabetes mellitus. Cases included 156 patients with diabetes mellitus and 156 controls without diabetes mellitus who attended in two public clinics in Durango City, Mexico. Sera of cases and controls were tested for the presence of anti- Toxoplasma IgG and IgM antibodies using commercially available enzyme-linked fluorescence assays (ELFA). Anti- T. gondii IgG antibodies were found in 10 (6.4%) of the 156 cases and in five (3.2%) of the 156 controls (odds ratio (OR): 2.06; 95% confidence interval (CI): 0.69 - 6.19; P = 0.18). The frequency of high (> 150 IU/mL) anti- T. gondii IgG levels in seropositive cases (1/10: 10.0%) was comparable to the one (1/5: 20%) in seropositive controls (OR: 0.44; 95% CI: 0.02 - 9.03; P = 1.00). None of the 10 cases and five controls with seropositivity to anti- T. gondii IgG antibodies were positive for anti- T. gondii IgM antibodies. Stratification by gender showed similar frequencies of T. gondii infection in female cases (7/107: 6.5%) and female controls (4/107: 3.7%) (OR: 1.80; 95% CI: 0.51 - 6.34; P = 0.53), and in male cases (3/49: 6.1%) and male controls (1/49: 2.0%) (OR: 3.13; 95% CI: 0.31 - 31.19; P = 0.61). We conclude that there is not serological evidence of an association between T. gondii infection and diabetes mellitus in the studied subjects in Durango City, Mexico. Further studies to elucidate the role of T. gondii in diabetes should be conducted.

Antibody reaction of human anti-Toxoplasma gondii positive and negative sera with Neospora caninum antigens

OpenAIRE

Nam, Ho-Woo; Kang, Seung-Won; Choi, Won-Young

1998-01-01

Anti-Neospora caninum antibody was detected in anti-Toxoplasma gondii positive and negative human sera by ELISA, western blot and immunofluorescence assay (IFA). Twelve cases out of 172 (6.7%) Toxoplasma-positive sera cross-reacted with both T. gondii and N. caninum antigens, and one out of 110 Toxoplasma-negative sera reacted with N. caninum antigen by ELISA. By western blot, all 12 sera reacted with T. gondii antigens with various banding patterns but specifically at 30 kDa (SAG1) and 22 kD...

A comparative study of small RNAs in Toxoplasma gondii of distinct genotypes

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Wang Jielin

2012-09-01

Full Text Available Abstract Background Toxoplasma gondii is an intracellular parasite with a significant impact on human health. Inside the mammalian and avian hosts, the parasite can undergo rapid development or remain inactive in the cysts. The mechanism that regulates parasite proliferation has not been fully understood. Small noncoding RNAs (sncRNA such as microRNAs (miRNAs are endogenous regulatory factors that can modulate cell differentiation and development. It is anticipated that hundreds of miRNAs regulate the expression of thousands of genes in a single organism. SncRNAs have been identified in T. gondii, however the profiles of sncRNAs expression and their potential regulatory function in parasites of distinct genotypes has largely been unknown. Methods The transcription profiles of miRNAs in the two genetically distinct strains, RH and ME49, of T. gondii were investigated and compared by a high-through-put RNA sequencing technique and systematic bioinformatics analysis. The expression of some of the miRNAs was confirmed by Northern blot analysis. Results 1,083,320 unique sequences were obtained. Of which, 17 conserved miRNAs related to 2 metazoan miRNA families and 339 novel miRNAs were identified. A total of 175 miRNAs showed strain-specific expression, of which 155 miRNAs were up-regulated in RH strain and 20 miRNAs were up-regulated in ME49 strain. Strain-specific expression of miRNAs in T. gondii could be due to activation of specific genes at different genomic loci or due to arm-switching of the same pre-miRNA duplex. Conclusions Evidence for the differential expression of miRNAs in the two genetically distinct strains of T. gondii has been identified and defined. MiRNAs of T. gondii are more species-specific as compared to other organisms, which can be developed as diagnostic biomarkers for toxoplasmosis. The data also provide a framework for future studies on RNAi-dependent regulatory mechanisms in the zoonotic parasite.

Molecular cloning and characterization of glucose transporter 1 ...

African Journals Online (AJOL)

Glucose transporter type-1 (glut1) and citrate synthase plays crucial role in glucose transport and regulation of tricarboxylic acid cycle (TCA) cycle in mammalian energy metabolism. The present study was aimed to clone and characterize glut1 and citrate synthase cDNA in water buffalo (Bubalus bubalis). Total of 90 ...

Screening of chemical compound libraries identified new anti-Toxoplasma gondii agents.

Science.gov (United States)

Adeyemi, Oluyomi Stephen; Sugi, Tatsuki; Han, Yongmei; Kato, Kentaro

2018-02-01

Toxoplasma gondii is the etiological agent of toxoplasmosis, a common parasitic disease that affects nearly one-third of the human population. The primary infection can be asymptomatic in healthy individuals but may prove fatal in immunocompromised individuals. Available treatment options for toxoplasmosis patients are limited, underscoring the urgent need to identify and develop new therapies. Non-biased screening of libraries of chemical compounds including the repurposing of well-characterized compounds is emerging as viable approach to achieving this goal. In the present investigation, we screened libraries of natural product and FDA-approved compounds to identify those that inhibited T. gondii growth. We identified 32 new compounds that potently inhibit T. gondii growth. Our findings are new and promising, and further strengthen the prospects of drug repurposing as well as the screening of a wide range of chemical compounds as a viable source of alternative anti-parasitic therapeutic agents.

Neospora caninum and Toxoplasma gondii antibody prevalence in Alaska wildlife.

Science.gov (United States)

Stieve, Erica; Beckmen, Kimberlee; Kania, Stephen A; Widner, Amanda; Patton, Sharon

2010-04-01

Free-ranging caribou and moose populations in some regions of Alaska undergo periodic declines in numbers. Caribou and moose are managed by the state as valuable resources for not only sustenance and subsistence, but also for cultural heritage. Incidence and prevalence of diseases that may impact herd health and recruitment from year to year are relevant to management decisions aimed to protect the long-term viability of these herds. Neospora caninum and Toxoplasma gondii are two apicomplexan parasites that can cause neurologic disease and abortions in their intermediate hosts and less frequently cause disease in their definitive hosts. The definitive hosts of N. caninum and T. gondii are canids and felids, respectively, and prevalence in the environment is in part dependent on maintenance of the life cycle through the definitive hosts. Serum samples from caribou (Rangifer tarandus, n=453), wolf (Canis lupus, n=324), moose (Alces alces, n=201), black-tailed deer (Odocoileus hemionus, n=55), coyote (Canis latrans, n=12), and fox (Vulpes vulpes, n=9) collected in Alaska were assayed for N. caninum- and T. gondii-reactive antibodies with an immunofluorescent antibody test (IFAT) and a modified agglutination test (MAT), respectively. Seroprevalence of N. caninum was greater in caribou (11.5%) than in wolves (9.0%), moose (0.5%), or black-tailed deer (0%). Seroprevalence of T. gondii was greater in wolves (17.8%) than in caribou (0.4%), moose (0%), or black-tailed deer (0%). Seroprevalence of N. caninum and T. gondii were 16.7% and 0.0% in coyotes and 0.0% and 12.5% in fox, but small sample sizes prevented further analysis. Antibodies to N. caninum in young caribou compared to adult caribou suggest that vertical transmission may be an important component of new infections in Alaskan caribou. The spatial distribution of antibody-positive individuals across Alaska may reflect differences in frequency of definitive hosts and alteration of predation patterns among regions.

Molecular cloning and expression of the gene encoding the kinetoplast-associated type II DNA topoisomerase of Crithidia fasciculata.

Science.gov (United States)

Pasion, S G; Hines, J C; Aebersold, R; Ray, D S

1992-01-01

A type II DNA topoisomerase, topoIImt, was shown previously to be associated with the kinetoplast DNA of the trypanosomatid Crithidia fasciculata. The gene encoding this kinetoplast-associated topoisomerase has been cloned by immunological screening of a Crithidia genomic expression library with monoclonal antibodies raised against the purified enzyme. The gene CfaTOP2 is a single copy gene and is expressed as a 4.8-kb polyadenylated transcript. The nucleotide sequence of CfaTOP2 has been determined and encodes a predicted polypeptide of 1239 amino acids with a molecular mass of 138,445. The identification of the cloned gene is supported by immunoblot analysis of the beta-galactosidase-CfaTOP2 fusion protein expressed in Escherichia coli and by analysis of tryptic peptide sequences derived from purified topoIImt. CfaTOP2 shares significant homology with nuclear type II DNA topoisomerases of other eukaryotes suggesting that in Crithidia both nuclear and mitochondrial forms of topoisomerase II are encoded by the same gene.

Molecular cloning of transcripts induced by UV-radiation in rodent cells

International Nuclear Information System (INIS)

Fornace, A.J. Jr.; Mitchell, J.B.

1987-01-01

Several inducible DNA repair genes have been well characterized in bacteria. In eukaryotes including mammalian cells, there is increasing evidence that similar events may occur. Recently, the authors have shown that hybridization subtraction can be used to enrich for sequences induced only several fold by a particular cell treatment such as heat shock. Chinese hamster V79 cells were UV-irradiated with 17 Jm/sup -2/ and cDNA was synthesized from the polyadenylated (poly A) RNA. This ''UV'' cDNA was hybridized with a 3 fold excess of polyA RNA from unirradiated cells and the nonhybridizing cDNA was isolated. With this approach, UV-induced sequences were enriched over 20 fold. This enriched cDNA was cloned into a high copy number plasmid and a cDNA library was constructed. By RNA dot blot and northern analysis, 42 clones from this library were found to represent transcripts induced 3 to 25 fold by UV. The most common isolates were found to be metallothionein transcripts by DNA sequencing. The metallothionein transcripts were found to be induced 10 to 25 fold by UV with maximum induction at 4-8 h after 10 Jm/sup -2/. A similar approach was also used with a Chinese hamster ovary line which does not express metallothionein and multiple clones were isolated which represented transcripts induced 3-15 fold by UV. Except for the metallothionein clones, the other Chinese hamster cDNA clones have not been identified, but it is probable that the protein products of at least some of these transcripts play a role in the cellular response to UV damage

Sporulation and survival of Toxoplasma gondii oocysts in different types of commercial cat litters

Science.gov (United States)

Toxoplasma gondii oocysts are environmentally resistant and can survive outdoors for months in the dry and cold climates. In the present study, sporulation and survival of T. gondii oocysts was studied in different types of cat litters commercially available in the US. Oocysts sporulated within 2-...

Epidemiology and Molecular Prevalence of Toxoplasma gondii in Cattle Slaughtered in Zahedan and Zabol Districts, South East of Iran

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Davood ANVARI

2018-03-01

Full Text Available Background: Toxoplasma gondii is an obligate, intracellular parasite which causes the toxoplasmosis in humans and warm-blooded animals. Red meat is an important source for transmission of the infection to humans. This study aimed to determine the prevalence of Toxoplasma among imported and indigenous cattle in the Sistan region.Methods: One hundred samples from slaughtered cattle were collected from two abattoirs of Zabol and Zahedan, South East of Iran in 2015. Each sample was a mixture of three muscle, including tongue, cardiac, and triceps. Additional data of each cattle, including sex, breed, age, indigenous or imported, location of slaughter, management practices, and feeding system were obtained through observations and interviews. Infection by T. gondii was determined by PCR method.Results: The prevalence of Toxoplasma in indigenous cattle was 6% and in imported cattle was 26%, and this difference was statistically significant (P=0.006. Moreover, the prevalence of Toxoplasma was statistically associated with management practices (P=0.01 and feeding system (P=0.001. However, relationship between the prevalence of Toxoplasma with age, breed, sex, and location of slaughter was not statistically significant.Conclusion: Since the prevalence of Toxoplasma among imported cattle is higher than indigenous cattle, so strict supervision for importing livestock from neighboring countries is necessary.

Toxoplasmosis in dogs: first report of Toxoplasma gondii infection in any animal species in Angola.

Science.gov (United States)

Lopes, Ana Patrícia; Granada, Sara; Oliveira, Ana Cristina; Brancal, Hugo; Dubey, Jitender P; Cardoso, Luís; Vilhena, Hugo

2014-10-01

Despite the worldwide importance of zoonotic parasite Toxoplasma gondii, nothing is known of toxoplasmosis in animals in Angola. The present study aimed at estimating the seroprevalence and also at assessing correlates of T. gondii infection in pet dogs from Luanda, Angola. Dogs (n = 103) brought to a veterinary clinic in the city of Luanda were investigated. Serum samples were tested for antibodies to T. gondii with a modified agglutination test (MAT) commercial kit, at serial dilutions of 1∶20 to 1∶160. In accordance with the established cutoff value (MAT ≧20), 16 dogs [15·5%; 95% confidence interval (CI): 9·2-24·0%] had antibodies to T. gondii: 10 had a titer of 20, two had a titer of 40, and four had a titer of 80. Age (≧12 months) was found to be a risk factor for infection [odds ratio (OR) = 9·23; 95% CI: 1·16-73·27). For each 1-year increase in age, the risk of a dog being found seropositive significantly increased by an OR of 1·18 (95% CI: 1·02-1·36). The present study, which represents the first serological survey of T. gondii in any animal species from Angola, reveals a 15·5% seroprevalence of infection in pet dogs in Luanda. Further studies are needed to better understand the epidemiology of zoonotic T. gondii infection in Luanda and also in Angola.

Occurrence of Toxoplasma gondii antibodies in lowland tapirs maintained ex situ in Brazil and Paraguay

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Maria Fernanda Naegeli Gondim

Full Text Available ABSTRACT: Lowland Tapir ( Tapirus terrestris is the second largest South American land mammal. It is strictly herbivorous and its exposure to Toxoplasma gondii should be indicative of environmental contamination by oocysts.In the present study antibodies to T. gondii in 47 Brazilian tapirs maintained ex situ in 10 Brazilian and in one Paraguayan institution were sought in serum samples by the modified agglutination test (MAT ≥25. None of the animals presented clinical signs during the study. From 47 animals 35 (74.5% were positive with titers of 25 in 8, 50 in 6, 100 in 12, 200 in 5, 400 in 1 and 800 in 3. One animal had samples collected on twice, and 19 were born in captivity. There was no association between occurrence of T. gondii antibodies and gender, and positive animals were reported in all institutions. The high occurrence of seropositive tapirs born ininstitutions (54.3% confirmed the high exposure of these mammals to T. gondii in captivity. Only two cases ofabortion were reported, but it was not possible to correlate these abortions to T. gondii infection.

Molecular cloning and characterization of duck interleukin-17

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Interleukin-17 (IL-17) belonging to the Th17 family is a proinflammatory cytokine produced by activated T cells. A 1034-bp cDNA encoding duck IL-17 (duIL-17) was cloned from ConA-activated splenic lymphocytes of ducks. The encoded protein, predicted to consisted of 169 amino acids, displayed a molec...

Construction and Rescue of a Molecular Clone of Deformed Wing Virus (DWV.

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Benjamin Lamp

Full Text Available European honey bees are highly important in crop pollination, increasing the value of global agricultural production by billions of dollars. Current knowledge about virulence and pathogenicity of Deformed wing virus (DWV, a major factor in honey bee colony mortality, is limited. With this study, we close the gap between field research and laboratory investigations by establishing a complete in vitro model for DWV pathogenesis. Infectious DWV was rescued from a molecular clone of a DWV-A genome that induces DWV symptoms such as crippled wings and discoloration. The expression of DWV proteins, production of infectious virus progeny, and DWV host cell tropism could be confirmed using newly generated anti-DWV monoclonal antibodies. The recombinant RNA fulfills Koch's postulates circumventing the need of virus isolation and propagation of pure virus cultures. In conclusion, we describe the development and application of a reverse genetics system for the study of DWV pathogenesis.

Cytomegalovirus and Toxoplasma gondii seroprevalence in a Brazilian liver transplant waiting

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R. A. M. B. Almeida

2007-01-01

Full Text Available Cytomegalovirus (CMV disease is a major cause of morbidity and mortality in solid organ transplantation. Disseminated toxoplasmosis after liver transplantation is a rare but fatal event. Serologic screening of the donor and the recipient is essential to prophylactic management, early diagnosis and therapeutic strategies to minimize the consequences of these infections. The aim of the present study was to determine the seroprevalence of CMV and Toxoplasma gondii (TG in a Brazilian liver transplant waiting list (LTWL. Serological data were collected from 44 candidates on the LTWL between May 2003 and November 2004. Serological investigation of antibodies IgM and IgG against CMV (anti-CMV and TG (anti-T. gondii was performed using fluorometry commercial kits. IgG anti-CMV was positive in 37 patients (94.9% out of 39 available results. There were not IgM anti-CMV positive results. Out of 36 analyzed patients, 22 (61.1% presented positive IgG anti-T. gondii and none had positive IgM anti-T. gondii. The high CMV seroprevalence among our LTWL reinforces the need for appropriate protocols to avoid related complications, like reactivation and superinfection by CMV. Environmental and drug prophylactic strategies against primary infection and reactivation, as well as early diagnosis and treatment of toxoplasmosis complications, are essential for the good outcome of transplant patients.

Detecção de anticorpos anti-Toxoplasma gondii em suínos criados e abatidos no Estado da Bahia, Brasil Detection anti-Toxoplasma gondii antibodies in swines bred and abated in the Bahia State, Brazil

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Rodrigo A. Bezerra

2009-09-01

Full Text Available Objetivou-se verificar a ocorrência de anticorpos anti-Toxoplasma gondii em suínos criados e abatidos no Estado da Bahia. Foram coletadas e examinadas 465 amostras de sangue de suínos provenientes de criações de diferentes locais desse estado. Para a pesquisa de anticorpos anti-T. gondii, foi utilizada a técnica de Imunoadsorção Enzimática (ELISA e considerados positivos todos os animais com títulos iguais ou maiores que 1:16. Desses, 18,27% (85/465 foram positivos para anticorpos anti-T. gondii, sendo 30,76% (24/78 em Ilhéus, 18,10% (21/116 em Itabuna e 14,76% (40/271 em Simões Filho. Foram observadas diferenças significativas quanto ao sexo dos animais (p = 0,0171, ao sistema de criação (p = 0,0002 e à procedência dos animais (p = 0,0278 no município de Itabuna. Anticorpos anti-T. gondii foram encontrados nos animais estudados, podendo ser estes animais fonte de infecção para a população humana local.This study was performed to verify the occurrence of anti-Toxoplasma gondii antibodies in swine raised and slaughtered in the state of Bahia, Brazil. Four hundred sixty five swine blood samples from farms of different cities had been collected and examined. Anti-T. gondii antibodies was detected by the enzyme-linked immunosorbent assay (ELISA and considered positive all the animals with equal or bigger headings than 1:16. From these, 18.27% (85/465 of total sample were positive for T. gondii, 30.76% (24 in Ilhéus, 18.10% (21/116 in Itabuna and 14.76% (40/271 in Simões Filho. Significant differences were observed regarding animal sex (p = 0.0171, raising system (p = 0.0002 and origin of the animals (p = 0.0278 in the city of Itabuna. The occurrence of anti-T. gondii antibodies shows that swine can be a source of infection for the local human population.

Genotyping of samples from German patients with ocular, cerebral and systemic toxoplasmosis reveals a predominance of Toxoplasma gondii type II.

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Herrmann, Daland C; Maksimov, Pavlo; Hotop, Andrea; Groß, Uwe; Däubener, Walter; Liesenfeld, Oliver; Pleyer, Uwe; Conraths, Franz J; Schares, Gereon

2014-10-01

Toxoplasmosis is an important zoonosis transmitted from animals to humans world-wide. In order to determine Toxoplasma gondii genotypes in individuals living in Germany and to compare findings with those in animals, we analysed nine independent and unlinked genetic markers (nSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) by PCR-RFLP in 83 archived T. gondii-positive DNA samples from patients with ocular toxoplasmosis (n=35), toxoplasmic encephalitis (n=32), systemic toxoplasmosis after bone-marrow transplantation (n=15) and congenital toxoplasmosis (n=1). In 46 of these 83 samples the presence of T. gondii DNA was confirmed by conventional end-point PCR. Among these, 17 T. gondii-positive samples were typed at all nine loci. The majority (15/17, 88.2%) of these samples were of T. gondii type II (i.e., including both, the Apico type II and Apico type I variants). In addition, in one sample a T. gondii type II/type III allele combination and in another sample a T. gondii genotype displaying type III alleles at all markers was observed. In the remaining 11 samples, in which T. gondii could only be partially typed, exclusively type II (n=10) or type III (n=1) alleles were observed. Results of the present study suggest that the majority of patients in Germany are infected with type II T. gondii regardless of the clinical manifestation of toxoplasmosis. This finding is in accord with the predominance of type II T. gondii in oocysts isolated from cats and in tissues of other intermediate hosts in Germany. Copyright © 2014 Elsevier GmbH. All rights reserved.

[The seroprevalence of Toxoplasma gondii in women from Sanliurfa, a province with a high raw meatball consumption].

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Tekay, Fikret; Ozbek, Erdal

2007-01-01

Toxoplasma gondii is an obligate intracellular protozoan that can infect all kind of birds and all mammals including humans and is common throughout the world. The prevalence varies according to social and cultural habits, pet cats in homes and geographic factors. Domestic cats are considered to be an important source of Toxoplasma gondii infection. From January to June 2006, the prevalence of toxoplasmosis was retrospectively monitored from blood samples that had been sent to our laboratory in order to determine the levels of IgM and IgG. All the subjects were women and 2,586 blood samples were investigated with the chemiluminescence immunoassay method. The rates of Toxoplasma gondii IgM antibodies were found to be 3.0% (78/2,586) and that of Toxoplasma gondii IgG antibodies, 69.5% (1.798/2,586). The total rate of positivity of Toxoplasma gondii antibodies was 69.6% (1,801/2,586) and the negativity, 30.4% (785/2,586). The highest positive rates have been reported in the southeastern region of Turkey and the 69.6% detected in our study seems to be the highest rate. Raw meatball consumption is common in our region and raw meat has a high risk of Toxoplasma gondii infection by direct ingestion of tissue cysts. As a result we consider that the high frequency of Toxoplasma gondii seropositivity in this region is due to raw meatball consumption.

Detection of Toxoplasma gondii in Diabetic Patients Using the Nested PCR Assay via RE and B1 Genes.

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Mousavi, Mohammad; Saravani, Ramin; Jafari Modrek, Mohammad; Shahrakipour, Mahnaz; Sekandarpour, Sina

2016-02-01

Toxoplasma gondii is an obligate intracellular protozoan parasite that exists worldwide. Various techniques have been developed for T. gondii detection. The aim of this study was the detection of T. gondii in diabetic patients with RE and B1 genes and the comparison of these two genes for diagnosis using the nested-PCR assay method. DNA samples from 205 diabetic patients who had been referred to the diabetes center of Ali Asghar hospital in Zahedan, Iran, were collected and analyzed using the nested-PCR assay method. Toxoplasma antibody data gathered using the enzyme-linked immunosorbent assay (ELISA) method from a previous study was used to group patients. The data were analyzed using SPSS 18. The chi-square test was used for comparison. Of the diabetic patients selected, the following results were obtained: 53 (IgG+, IgM+); 20 (IgG-, IgM+); 72 (IgG+, IgM-); and 60 (IgG-, IgM-). The nested-PCR detected the following: in the acute group, 21/53 (39.63%), 30/53 (56.60%) (IgM+, IgG+); in the chronic group, 40/72 (55.56%), 51/72 (70.83%), (IgG+, IgM-); in the false positive group, 18/20 (90%), 17/20 (85%) (IgM+, IgG-); and sero-negative samples of 38/60 (63.33%) and 60/ 41 (77.35%) for RE and B1 genes, respectively. The prevalence of toxoplasmosis showed positive in patients with diabetes in the B1 gene 139 (67.8%) and RE gene 117 (57.1%). Our study demonstrated that the B1 gene, more so than the RE gene, showed positive samples and can be used to detect toxoplasmosis, although the B1 gene, in comparison to the RE gene, did not show any superiority of molecular diagnosing capability. Results also showed that toxoplasma molecular detection methods can be used instead of routine serological detection methods in a clinical laboratory testing.

The neurotropic parasite Toxoplasma gondii increases dopamine metabolism.

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Emese Prandovszky

Full Text Available The highly prevalent parasite Toxoplasma gondii manipulates its host's behavior. In infected rodents, the behavioral changes increase the likelihood that the parasite will be transmitted back to its definitive cat host, an essential step in completion of the parasite's life cycle. The mechanism(s responsible for behavioral changes in the host is unknown but two lines of published evidence suggest that the parasite alters neurotransmitter signal transduction: the disruption of the parasite-induced behavioral changes with medications used to treat psychiatric disease (specifically dopamine antagonists and identification of a tyrosine hydroxylase encoded in the parasite genome. In this study, infection of mammalian dopaminergic cells with T. gondii enhanced the levels of K+-induced release of dopamine several-fold, with a direct correlation between the number of infected cells and the quantity of dopamine released. Immunostaining brain sections of infected mice with dopamine antibody showed intense staining of encysted parasites. Based on these analyses, T. gondii orchestrates a significant increase in dopamine metabolism in neural cells. Tyrosine hydroxylase, the rate-limiting enzyme for dopamine synthesis, was also found in intracellular tissue cysts in brain tissue with antibodies specific for the parasite-encoded tyrosine hydroxylase. These observations provide a mechanism for parasite-induced behavioral changes. The observed effects on dopamine metabolism could also be relevant in interpreting reports of psychobehavioral changes in toxoplasmosis-infected humans.

Toxoplasma gondii seroprevalence in free-ranging moose (Alces alces) hunted for human consumption in Estonia

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Remes, Noora; Kärssin, Age; Must, Kärt

2018-01-01

In Estonia, northeastern Europe, antibodies against Toxoplasma gondii are common in many host species, including wildlife, domestic animals, and humans. Our nationwide study aimed to estimate T. gondii seroprevalence and its geographical distribution, and to evaluate plausible risk factors for se...

Molecular cloning, characterization and functional analysis of a 3 ...

African Journals Online (AJOL)

STORAGESEVER

2009-08-04

Aug 4, 2009 ... other plant HMGRs and contained 2 transmembrane domains and a catalytic domain. The potential significance ... used as animal feed. Therefore ... Table 1. Primers used in the cloning and analysis of JcHMGR gene. Primers.

Mechanics of the Toxoplasma gondii oocyst wall

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The ability of microorganisms to survive under extreme conditions is closely related to the physicochemical properties of their wall. In the ubiquitous protozoan parasite Toxoplasma gondii, the oocyst stage possesses a bilayered wall that protects the dormant but potentially infective parasites from...

Toxoplasma gondii actively inhibits neuronal function in chronically infected mice.

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Fahad Haroon

Full Text Available Upon infection with the obligate intracellular parasite Toxoplasma gondii, fast replicating tachyzoites infect a broad spectrum of host cells including neurons. Under the pressure of the immune response, tachyzoites convert into slow-replicating bradyzoites, which persist as cysts in neurons. Currently, it is unclear whether T. gondii alters the functional activity of neurons, which may contribute to altered behaviour of T. gondii-infected mice and men. In the present study we demonstrate that upon oral infection with T. gondii cysts, chronically infected BALB/c mice lost over time their natural fear against cat urine which was paralleled by the persistence of the parasite in brain regions affecting behaviour and odor perception. Detailed immunohistochemistry showed that in infected neurons not only parasitic cysts but also the host cell cytoplasm and some axons stained positive for Toxoplasma antigen suggesting that parasitic proteins might directly interfere with neuronal function. In fact, in vitro live cell calcium (Ca(2+ imaging studies revealed that tachyzoites actively manipulated Ca(2+ signalling upon glutamate stimulation leading either to hyper- or hypo-responsive neurons. Experiments with the endoplasmatic reticulum Ca(2+ uptake inhibitor thapsigargin indicate that tachyzoites deplete Ca(2+ stores in the endoplasmatic reticulum. Furthermore in vivo studies revealed that the activity-dependent uptake of the potassium analogue thallium was reduced in cyst harbouring neurons indicating their functional impairment. The percentage of non-functional neurons increased over time In conclusion, both bradyzoites and tachyzoites functionally silence infected neurons, which may significantly contribute to the altered behaviour of the host.

Cloning of Plasmodium falciparum by single-cell sorting.

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Miao, Jun; Li, Xiaolian; Cui, Liwang

2010-10-01

Malaria parasite cloning is traditionally carried out mainly by using the limiting dilution method, which is laborious, imprecise, and unable to distinguish multiply-infected RBCs. In this study, we used a parasite engineered to express green fluorescent protein (GFP) to evaluate a single-cell sorting method for rapidly cloning Plasmodium falciparum. By dividing a two-dimensional scattergram from a cell sorter into 17 gates, we determined the parameters for isolating singly-infected erythrocytes and sorted them into individual cultures. Pre-gating of the engineered parasites for GFP allowed the isolation of almost 100% GFP-positive clones. Compared with the limiting dilution method, the number of parasite clones obtained by single-cell sorting was much higher. Molecular analyses showed that parasite isolates obtained by single-cell sorting were highly homogenous. This highly efficient single-cell sorting method should prove very useful for cloning both P. falciparum laboratory populations from genetic manipulation experiments and clinical samples. Copyright 2010 Elsevier Inc. All rights reserved.

Cloning of Plasmodium falciparum by single-cell sorting

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Miao, Jun; Li, Xiaolian; Cui, Liwang

2010-01-01

Malaria parasite cloning is traditionally carried out mainly by using the limiting dilution method, which is laborious, imprecise, and unable to distinguish multiply-infected RBCs. In this study, we used a parasite engineered to express green fluorescent protein (GFP) to evaluate a single-cell sorting method for rapidly cloning Plasmodium falciparum. By dividing a two dimensional scattergram from a cell sorter into 17 gates, we determined the parameters for isolating singly-infected erythrocytes and sorted them into individual cultures. Pre-gating of the engineered parasites for GFP allowed the isolation of almost 100% GFP-positive clones. Compared with the limiting dilution method, the number of parasite clones obtained by single-cell sorting was much higher. Molecular analyses showed that parasite isolates obtained by single-cell sorting were highly homogenous. This highly efficient single-cell sorting method should prove very useful for cloning both P. falciparum laboratory populations from genetic manipulation experiments and clinical samples. PMID:20435038

Molecular Cloning Expression And Purification Studies With An ORF Of Mycobacterium Tuberculosis

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Chiranjibi Chaudhary

2017-08-01

Full Text Available The study was initiated to develop a recombinant strain for expression and production of large scale protein and to develop its purification protocol. The MRAORF-X was amplified from the genomic DNA of M. tuberculosis H37Ra. The amplicon was successfully cloned in a cloning vector pGEM-T Easy and transformed in cloning host DH5amp945. Recombinant clones were identified by blue-white screening and insert presence was confirmed by restriction digestion of plasmid isolated from white colonies. Expression vector pET32a was used for protein expression. The recombinant plasmid was transformed into expression host BL21 and protein expression was checked by SDS-PAGE. The desired protein was approximately 60 kDa in size including tags. The purification protocol was established for purification from inclusion bodies. The purity of purified protein was assessed by SDS-PAGE gel run and presence of a single band at 60 kDa suggested that the inclusion bodies were a good source of purified protein.

Diagnosis of pulmonary infection with Toxoplasma gondii in immunocompromised HIV-positive patients by real-time PCR

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Petersen, E.; Edvinsson, B.; Lundgren, Bettina

2006-01-01

. In positive samples, the genotype of the parasite was determined by sequence analysis of the GRA6 gene. Positive results were achieved for 2% (7/332) of the samples tested. Genotyping was possible in two samples and revealed GRA6 type II T. gondii. PCR for detecting T. gondii in BAL samples should...... be performed in all immunosuppressed HIV-positive patients with symptoms of a systemic infection of unknown etiology. Trimethoprim-sulfamethoxazole prophylaxis does not exclude concomitant infection with T. gondii....

Lacrimal secretory IgA in active posterior uveitis induced by Toxoplasma gondii

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Maria Isabel Lynch

2004-12-01

Full Text Available It is quite difficult to diagnose active toxoplasmosis in patients with ocular toxoplasmosis. Active posterior uveitis presumably due to Toxoplasma gondii infection (APUPT is seldom produced during a prime-infection; hence most patients do not show high IgM antibodies. High levels of IgA have been described in active toxoplasmosis. The purpose of this study was to investigate possible association between APUPT and the specific anti-parasite sIgA in tears. The study was carried out as case-control. Tears of 25 clinically confirmed APUPT patients and 50 healthy control subjects were analyzed. All were IgG seropositive. Specific sIgA was determined by ELISA assay using T. gondii RH strain crude extract. Anti-T. gondii sIgA was found in 84% of the cases and in 22% of the control subjects. The intensity of the reaction was higher in APUPT cases (P = 0.007. There was strong association between APUPT patients and lacrimal sIgA (odds-ratio 18.61, P = 0.0001. ELISA test sensitivity was 84% and specificity 78% . Our data suggest that anti-T.gondii secretory IgA found in tears may become an important marker for active ocular toxoplasmosis.

Toxoplasma gondii seroprevalence and genotype diversity in select wildlife species from the southeastern United States

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Richard W. Gerhold

2017-10-01

Full Text Available Abstract Background Toxoplasma gondii is a widespread protozoan parasite that infects humans and other animals. Previous studies indicate some genotypes of T. gondii are more frequently isolated in wildlife than agricultural animals, suggesting a wild/feral animal diversity model. To determine seroprevalence and genetic diversity of T. gondii in southeastern US wildlife, we collected sera from 471 wild animals, including 453 mammals and 18 birds, between 2011 and 2014. These serum samples were assayed for T. gondii infection using the modified agglutination test (MAT. Heart or tongue tissues from 66 seropositive animals were bioassayed in mice and 19 isolates were obtained. The isolated parasites were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP method employing 10 genetic markers. Results One hundred and ninety-six of 471 samples (41.6% had a titer ≥1:32 and were considered positive for T. gondii infection. Of 453 mammals, 195 (43% were seropositive, whereas only one (5.6% of 18 birds was seropositive. The seroprevalence in mammals was significantly higher than in the birds. Mammalian hosts with adequate samples size (≥ 20 comprised white-tailed deer (n = 241, feral hogs (n = 100, raccoons (n = 34 and coyotes (n = 22, with seroprevalences of 41.0%, 51.0%, 50.0% and 72.7%, respectively. Coyotes had significantly higher seroprevalence than the white-tailed deer. Genotyping revealed five distinct genotypes, including the ToxoDB PCR-RFLP genotype #5 (a.k.a type 12 for 15 isolates, genotype #3 (a.k.a. type II for 1 isolate, and genotypes #154, #167 and #216, each for 1 isolate. The results showed moderate to high infection rates of T. gondii in white-tailed deer, feral hogs, raccoons and coyotes. Genotyping results indicated limited genetic diversity and a dominance of genotype #5, which has been reported as a major type in wildlife in North America. Conclusions We conclude that T. gondii

Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests

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Aroussi Abdelkrim

2015-01-01

Full Text Available In France, some cases of severe toxoplasmosis have been linked to the consumption of horse meat that had been imported from the American continent where atypical strains of Toxoplasma gondii are more common than in Europe. Many seroprevalence studies are presented in the literature but risk assessment of T. gondii infection after horse meat consumption is not possible in the absence of validated serological tests and the unknown correlation between detection of antibodies against T. gondii and presence of tissue cysts. We performed magnetic-capture polymerase chain reaction (MC-PCR to detect T. gondii DNA in 231 horse meat samples purchased in supermarkets in France and evaluated the performance and level of agreement of the modified agglutination test (MAT and enzyme-linked immunosorbent assay (ELISA in the meat juices. The serological tests lacked sensitivity, specificity, and agreement between them, and there was no correlation with the presence of T. gondii DNA in horse meat, raising concerns about the reliability of T. gondii seroprevalence data in horses from the literature. T. gondii DNA was detected in 43% of horse meat samples but the absence of strain isolation in mice following inoculation of more than 100 horse meat samples suggests a low distribution of cysts in skeletal muscles and a low risk of T. gondii infection associated with horse meat consumption. However, to avoid any risk of toxoplasmosis, thorough cooking of horse meat is recommended.

Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests.

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Aroussi, Abdelkrim; Vignoles, Philippe; Dalmay, François; Wimel, Laurence; Dardé, Marie-Laure; Mercier, Aurélien; Ajzenberg, Daniel

2015-01-01

In France, some cases of severe toxoplasmosis have been linked to the consumption of horse meat that had been imported from the American continent where atypical strains of Toxoplasma gondii are more common than in Europe. Many seroprevalence studies are presented in the literature but risk assessment of T. gondii infection after horse meat consumption is not possible in the absence of validated serological tests and the unknown correlation between detection of antibodies against T. gondii and presence of tissue cysts. We performed magnetic-capture polymerase chain reaction (MC-PCR) to detect T. gondii DNA in 231 horse meat samples purchased in supermarkets in France and evaluated the performance and level of agreement of the modified agglutination test (MAT) and enzyme-linked immunosorbent assay (ELISA) in the meat juices. The serological tests lacked sensitivity, specificity, and agreement between them, and there was no correlation with the presence of T. gondii DNA in horse meat, raising concerns about the reliability of T. gondii seroprevalence data in horses from the literature. T. gondii DNA was detected in 43% of horse meat samples but the absence of strain isolation in mice following inoculation of more than 100 horse meat samples suggests a low distribution of cysts in skeletal muscles and a low risk of T. gondii infection associated with horse meat consumption. However, to avoid any risk of toxoplasmosis, thorough cooking of horse meat is recommended. © A. Aroussi et al., published by EDP Sciences, 2015.

Immune-mediated bone marrow failure syndromes of progenitor and stem cells: molecular analysis of cytotoxic T cell clones.

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Ramon Tiu

2007-03-01

Full Text Available The unique structure of the T cell receptor (TCR enables molecular identification of individual T cell clones and provides an unique opportunity for the design of molecular diagnostic tests based on the structure of the rearranged TCR chain e.g., using the TCR CDR3 region. Initially, clonal T cell malignancies, including T cell large granular lymphocyte leukemia (T-LGL, mucosis fungoides and peripheral T cell lymphoma were targets for the TCR-based analytic assays such as detection of clonality by T-gamma rearrangement using y-chain-specific PCR or Southern Blotting. Study of these disorders facilitated further analytic concepts and application of rational methods of TCR analysis to investigations of polyclonal T cell-mediated diseases. In hematology, such conditions include graft versus host disease (GvHD and immune-mediated bone marrow failure syndromes. In aplastic anemia (AA, myelodysplastic syndrome (MDS or paroxysmal nocturnal hemoglobinuria (PNH, cytotoxic T cell responses may be directed against certain antigens located on stem or more lineage-restricted progenitor cells in single lineage cytopenias. The nature of the antigenic targets driving polyclonal CTL responses remains unclear. Novel methods of TCR repertoire analysis, include VB flow cytometry, peptide-specific tetramer staining, in vitro stimulation assays and TCR CDR3-specific PCR. Such PCR assay can be either VB family-specific or multiplexed for all VB families. Amplified products can be characterized and quantitated to facilitate detection of the most immunodominant clonotypes. Such clonotypes may serve as markers for the global polyclonal T cell response. Identification of these clonotypes can be performed in blood and tissue biopsy material by various methods. Once immunodominant clonotypes corresponding to pathogenic CTL clones are identified they can serve as surrogate markers for the activity of the pathophysiologic process or even indicate the presence of specific

Characterization of the env gene and long terminal repeat of molecularly cloned Friend mink cell focus-inducing virus DNA.

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Adachi, A; Sakai, K; Kitamura, N; Nakanishi, S; Niwa, O; Matsuyama, M; Ishimoto, A

1984-01-01

The highly oncogenic erythroleukemia-inducing Friend mink cell focus-inducing (MCF) virus was molecularly cloned in phage lambda gtWES.lambda B, and the DNA sequences of the env gene and the long terminal repeat were determined. The nucleotide sequences of Friend MCF virus and Friend spleen focus-forming virus were quite homologous, supporting the hypothesis that Friend spleen focus-forming virus might be generated via Friend MCF virus from an ecotropic Friend virus mainly by some deletions. ...

Toxoplasma gondii in experimentally infected Bos taurus and Bos indicus semen and tissues Toxoplasma gondii em semen e tecidos de Bos taurus and Bos indicus experimentalmente infectados

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Leslie Scarpelli

2009-01-01

Full Text Available Eighteen young steers were inoculated with Toxoplasma gondii and randomly distributed into three groups of six animals each: GI, 2.5x10(5 "P" strain oocysts, GII, 5.0x10(6 "RH" strain tachyzoites, and GIII (Control. Clinical, serological and parasitemia exams were realized. Parasite investigation by bioassay and PCR was realized on semen and fragments of skeletal musculature, lymph nodes, brain, retina, spleen, liver, lung, testicle, epididymis and seminal vesicle. Blood and semen samples were collected on days -2, -1, 1, 3, 5, 7, 14 and weekly thereafter, up to postinfection day (PID 84. The inoculated steers (GI and GII presented hyperthermia from PID 3 to 16. Antibodies against T. gondii were detected through the indirect fluorescence antibody test (IFAT on PID 5 (1:16 in both inoculated groups (oocysts and tachyzoites, reaching peaks of 1:4096 on PID 7. Parasitemia outbursts occurred in all infected bovines, principally from PID 7 to 28, independent of the strain and inoculate used. Bioassays revealed the presence of parasites in semen samples of animals infected with oocysts (GI and tachyzoites (GII on several experimental days between PID 7 and 84. Tissue parasitism by T. gondii was diagnosed by bioassay and the PCR technique in several organ and tissue fragments. These findings suggest the possibility of sexual transmission of T. gondii in the bovine species.Dezoito bovinos foram inoculados com Toxoplasma gondii e distribuídos aleatoriamente em três grupos de seis bovinos cada: GI (2,5x10(5 oocistos da cepa "P", GII (5,0x10(6 taquizoítos da cepa "RH" e GIII (controle. Exames clínicos, sorológicos e parasitêmicos foram realizados. Pesquisas do parasito, por meio da bioprova e pela técnica de Reação em Cadeia pela Polimerase (PCR, foram realizadas no sêmen e em fragmentos de musculatura esquelética, linfonodos, cérebro, retina, baço, fígado, pulmão, testículo, epidídimo e vesícula seminal. Amostras de sangue e sêmen foram

Anticorpos contra Toxoplasma gondii em estudantes de medicina veterinária de Campo Grande, MS, Brasil Antibodies to Toxoplasma Gondii in veterinary medicine students of Campo Grande, MS, Brazil

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Flábio Ribeiro de Araújo

2000-12-01

Full Text Available Amostras de soro obtidas de estudantes do curso de Medicina Veteriná ria da Universidade para o Desenvolvimento do Estado e da Região do Pantanal, Campo Grande, MS, Brasil, foram examinadas para a presença de anticorpos contra Toxoplasma gondii. Dos 145 soros testados, 44 (30,34% foram positivos na hemaglutinação, com título igual ou superior a 1:16. Não foram observadas associações entre as caracterí sticas epidemiológicas examinadas, tais como hábitos alimentares (ingestão de carne bovina crua ou malpassada, vegetais crus/não lavados, produtos lácteos não pasteurizados ou contato constante com cães e a presença de anticorpos contra T. gondii, exceto pelo percentual significativamente maior de estudantes soropositivos que relataram ter contato freqüente com gatos (P=0,03.Serum samples obtained from students of the School of Veterinary Medicine, Universidade para o Desenvolvimento do Estado e da Região do Pantanal, Campo Grande, MS, Brazil, were examined for Toxoplasma gondii gondii antibodies. Serum samples from 44 (30.34% of the 145 veterinary students examined were positive on the hemagglutination test, yielding a titer of 1:16 or greater. There were no relations between the epidemiological characteristics examined, such as food habits (eating of raw or rare-cooked cattle meat, raw/uncleaned vegetables, unpasteurized dairy products, frequent contact with dogs and the presence of T. gondii antibodies, except for a significantly higher percentual of seropositive students that reported frequent contact with cats (P=0.03.

Toxoplasma gondii acetyl-CoA synthetase is involved in fatty acid elongation (of long fatty acid chains) during tachyzoite life stages.

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Dubois, David; Fernandes, Stella; Amiar, Souad; Dass, Sheena; Katris, Nicholas J; Botté, Cyrille Y; Yamaryo-Botté, Yoshiki

2018-06-01

Apicomplexan parasites are pathogens responsible for major human diseases such as toxoplasmosis caused by Toxoplasma gondii and malaria caused by Plasmodium spp. Throughout their intracellular division cycle, the parasites require vast and specific amounts of lipids to divide and survive. This demand for lipids relies on a fine balance between de novo synthesized lipids and scavenged lipids from the host. Acetyl-CoA is a major and central precursor for many metabolic pathways, especially for lipid biosynthesis. T. gondii possesses a single cytosolic acetyl-CoA synthetase ( Tg ACS). Its role in the parasite lipid synthesis is unclear. Here, we generated an inducible Tg ACS KO parasite line and confirmed the cytosolic localization of the protein. We conducted 13 C-stable isotope labeling combined with mass spectrometry-based lipidomic analyses to unravel its putative role in the parasite lipid synthesis pathway. We show that its disruption has a minor effect on the global FA composition due to the metabolic changes induced to compensate for its loss. However, we could demonstrate that Tg ACS is involved in providing acetyl-CoA for the essential fatty elongation pathway to generate FAs used for membrane biogenesis. This work provides novel metabolic insight to decipher the complex lipid synthesis in T. gondii . Copyright © 2018 by the American Society for Biochemistry and Molecular Biology, Inc.

Risk factors for acute Toxoplasma gondii diseases in Taiwan: a population-based case-control study.

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Ting-Yi Chiang

Full Text Available Although human toxoplasmosis is a notifiable disease in Taiwan since 2007, little is known about its risk factors. This study aimed to investigate the risk factors for acute Toxoplasma gondii diseases in Taiwan. We conducted a nationwide population-based case-control study. Cases of acute human toxoplasmosis notified to the Taiwan Centers for Diseases Control (Taipei, Taiwan during 2008-2013 were compared with controls that were randomly selected from healthy T. gondii-seronegative blood donors who participated in a nationwide T. gondii seroepidemiologic study during 2009-2010. Cases and controls were matched according to age, gender and residency at an 1:8 ratio. Structured questionnaires were used to gather information regarding risk factors. A total of 30 laboratory-confirmed acute T. gondii disease cases and 224 controls were enrolled. The most common clinical manifestation of the cases was flu-like symptoms (n = 20, followed by central nervous system disease (n = 4, ocular diseases (n = 3, abortion (n = 2, and congenital infection (n = 1. Multivariate conditional logistic regression showed that raw clam consumption (adjusted odds ratio [OR] = 3.7; 95% confidence interval [CI] = 1.4-9.9 and having a cat in the household (adjusted OR = 2.9; 95% CI = 1.1-7.9 were two independent risk factors for acute T. gondii disease. We conclude that raw shellfish consumption and domestic cat exposure were risk factors for acquiring acute T. gondii diseases in Taiwan. This finding may guide future research and control policies.

Latent Toxoplasma gondii infection leads to improved action control.

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Stock, Ann-Kathrin; Heintschel von Heinegg, Evelyn; Köhling, Hedda-Luise; Beste, Christian

2014-03-01

The parasite Toxoplasma gondii has been found to manipulate the behavior of its secondary hosts to increase its own dissemination which is commonly believed to be to the detriment of the host (manipulation hypothesis). The manipulation correlates with an up-regulation of dopaminergic neurotransmission. In humans, different pathologies have been associated with T. gondii infections but most latently infected humans do not seem to display overt impairments. Since a dopamine plus does not necessarily bear exclusively negative consequences in humans, we investigated potential positive consequences of latent toxoplasmosis (and the presumed boosting of dopaminergic neurotransmission) on human cognition and behavior. For this purpose, we focused on action cascading which has been shown to be modulated by dopamine. Based on behavioral and neurophysiological (EEG) data obtained by means of a stop-change paradigm, we were able to demonstrate that healthy young humans can actually benefit from latent T. gondii infection as regards their performance in this task (as indicated by faster response times and a smaller P3 component). The data shows that a latent infection which is assumed to affect the dopaminergic system can lead to paradoxical improvements of cognitive control processes in humans. Copyright © 2013 Elsevier Inc. All rights reserved.

The Ultrastructural Effects of Sulfachloropyrazine on Toxoplasma Gondii Tachyzoites

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YB Zeng

2013-03-01

Full Text Available Background: Toxoplasmosis is one of the most common parasitic infections of humans and other mammals. This study was aimed to understand the mechanism of action of veterinary medicine-sulfachlo­ropyrazine (SPZ, 99.97% against Toxoplasma gondii.Methods: T. gondii tachyzoites were soaked in PBS (as a control or SPZ (250 mg/mL for 2 h at 37 °C. After being processed, any ultrastructural changes of the tachyzoites that had occurred were observed by Scanning Electron Microscopy (SEM and Transmission Electron Microscopy (TEM.Results: The tachyzoites from control groups with a uniform size had a smooth surface and intact cell or nuclear membranes. In addition, an oval-shaped nucleus, conoids and micronemes were also observed. By contrast, many parasites from the SPZ-treated groups were detrimentally affected by the treatment. Some appeared to be of the vacuolization in their cytoplasm, with the substantial reduc­tion in the number of dense granules and the blur of some organelles.Conclusion: The morphology and ultrastructure of tachyzoites can be affected significantly by SPZ, which might kill the parasite by inhibiting its energy metabolism, inducing apoptosis and damaging its structure. The study provides an experimental basis for further study on the mechanism of SPZ against T. gondii.

Identification of cDNA clones expressing immunodiagnostic antigens from Trichinella spiralis

International Nuclear Information System (INIS)

Zarlenga, D.; Gamble, H.R.

1987-01-01

A cDNA expression library was built in lambda gt11 phage using poly A mRNA isolated from Trichinella spiralis muscle stage larvae. This library was screened with rabbit antibodies to parasite excretory-secretory (ES) products and greater than 180 clones were isolated. Thirteen clones producing highly immunogenic protein antigens were plaque purified and rescreened with pig antisera to T.spiralis, Trichuris suis or Ascaris suum to identify clones producing epitopes specific to T.spiralis ES products, only. Two clones, TsAc-2 and TsAc-8, which displayed strong interactions with pig antisera to T. spiralis were lysogenized in E. coli Y1089 and the protein extracted. Western blots of the crude fusion proteins revealed molecular weights of 133 kD and 129 kD, respectively. Northern blot analysis of total RNA with 32 P labelled cDNA:lambda gt11 probes indicated single RNA transcripts for each clone with molecular sizes corresponding to 800-850 nucleotides. dscDNA inserts were estimated by southern blot analysis to be 500 bp and 340 bp, respectively, with no cross-hybridization observed between the cloned sequences. Dot blots using pig sera to screen crude fusion protein preparations, total bacterial protein (negative controls) and crude worm extract or ES products from T.spiralis, T.suis and A.suum (positive controls) corroborated the specificity and sensitivity of these clones as potential diagnostic antigens for swine trichinellosis

Toxoplasma gondii seroprevalence in pet cats in Norway and risk factors for seropositivity.

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Sævik, Bente Kristin; Krontveit, Randi Ingebjørg; Eggen, Kristine P; Malmberg, Nina; Thoresen, Stein I; Prestrud, Kristin W

2015-12-01

The aims of the study were to estimate Toxoplasma gondii seroprevalence in pet cats in Norway and to evaluate risk factors for seropositivity. Additionally, serum biochemistry and haematological variables for T gondii seropositive and seronegative cats were compared. A convenience sample of surplus sera submitted to the Central Laboratory, Norwegian University of Life Sciences, was collected. The samples were from healthy cats and cats with a variety of diseases. Analyses for IgG antibodies to T gondii were performed with a commercial direct agglutination test, with 1:40 as the threshold value. For risk factor analysis a logistic regression model of the relationship between predictors and the outcome was applied. One hundred and ninety-six of 478 cats were seropositive for T gondii, and the estimated seroprevalence in the study sample was 41.0% (95% confidence interval 36.6-45.4). Compared with domestic cats, pedigree cats had reduced risk for Toxoplasma seropositivity (odds ratio [OR] 0.42). Males had increased risk (OR 1.63) compared with females. The effect of age was highly significant, and an increase in the cats' age across the interquartile range (IQR; 52-160 months/4-13 years of age) doubled the risk of Toxoplasma seropositivity (OR 2.11). The risk for Toxoplasma seropositivity among cats living in Oslo was significantly reduced (OR 0.51) when compared with the rest of Norway. Pet cats in Norway appear to be commonly exposed to T gondii. Signalment and geographical region influenced the odds of Toxoplasma seropositivity, whereas health status did not. © ISFM and AAFP 2015.

Stray dogs as indicators of Toxoplasma gondii distributed in the environment: the first report across an urban-rural gradient in China

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Yan Chao

2012-01-01

Full Text Available Abstract Background Toxoplasmosis is an important parasitic zoonosis caused by the protozoan Toxoplasma gondii that is distributed world-wide and infects a variety of hosts. However, the prevalence of T. gondii in the environment (such as soil, water and food is largely unknown. Due to the technical difficulty in oocyst counting directly, an alternative assay using the serologic status of T. gondii in free-living animals, such as stray or free-living dogs, as an indicator, can be used to evaluate environmental contamination indirectly, as they are exposed to the same risk of infection as humans and other animals. Results In the present study, 231 stray or free-living dogs across an urban-rural gradient were examined to assess the frequency of T. gondii in the environment. Specific antibodies to T. gondii were found in 93 dogs (40.3% by enzyme-linked immunosorbent assay (ELISA, and no statistically significant differences were observed in seroprevalences of T. gondii between urban dogs (38.7% and rural dogs (41% (p > 0.05. Conclusions A high seroprevalence of T. gondii in stray or free-living dogs in the present study indicates that there would be a wide distribution and a constant infection pressure of T. gondii across an urban-rural gradient, and the oocysts of T. gondii in the environment would be an important source of infection for humans and other animals both in urban and rural areas in China.

Presence of Toxoplasma gondii in Drinking Water from an Endemic Region in Southern Mexico.

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Hernandez-Cortazar, Ivonne B; Acosta-Viana, Karla Y; Guzman-Marin, Eugenia; Ortega-Pacheco, Antonio; Segura-Correa, Jose C; Jimenez-Coello, Matilde

2017-05-01

Toxoplasmosis can be acquired through the ingestion of contaminated drinking water with oocysts of Toxoplasma gondii, highly resistant to the routinely disinfection processes; based on chlorination commonly used in the water supply industry. The aim of this study was to determine the presence of T. gondii DNA in samples of public drinking water from an endemic region of southern Mexico. In total 74 samples of water (5 L each) were collected from the three well fields (I, II, and III) and 71 independent wells, distributing public drinking water to the city of Merida Yucatan, after passing through the chlorination process. Water samples were filtered and concentrated by a sucrose solution, then DNA was extracted and evaluated through a nested-PCR (nPCR) specific for T. gondii. Positive samples were detected in 5.4% (4/74) of the water samples. This is the first report of the presence of T. gondii DNA in public drinking water from a large city in southern Mexico, where their consumption without any postpurification treatment could pose a risk for acquiring the infection in the urban population.

An experimental Toxoplasma gondii dose response challenge model to study therapeutic or vaccine efficacy in cats.

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Jan B W J Cornelissen

Full Text Available High numbers of Toxoplasma gondii oocysts in the environment are a risk factor to humans. The environmental contamination might be reduced by vaccinating the definitive host, cats. An experimental challenge model is necessary to quantitatively assess the efficacy of a vaccine or drug treatment. Previous studies have indicated that bradyzoites are highly infectious for cats. To infect cats, tissue cysts were isolated from the brains of mice infected with oocysts of T. gondii M4 strain, and bradyzoites were released by pepsin digestion. Free bradyzoites were counted and graded doses (1000, 100, 50, 10, and 250 intact tissue cysts were inoculated orally into three cats each. Oocysts shed by these five groups of cats were collected from faeces by flotation techniques, counted microscopically and estimated by real time PCR. Additionally, the number of T. gondii in heart, tongue and brains were estimated, and serology for anti T. gondii antibodies was performed. A Beta-Poisson dose-response model was used to estimate the infectivity of single bradyzoites and linear regression was used to determine the relation between inoculated dose and numbers of oocyst shed. We found that real time PCR was more sensitive than microscopic detection of oocysts, and oocysts were detected by PCR in faeces of cats fed 10 bradyzoites but by microscopic examination. Real time PCR may only detect fragments of T. gondii DNA without the presence of oocysts in low doses. Prevalence of tissue cysts of T. gondii in tongue, heart and brains, and anti T. gondii antibody concentrations were all found to depend on the inoculated bradyzoite dose. The combination of the experimental challenge model and the dose response analysis provides a suitable reference for quantifying the potential reduction in human health risk due to a treatment of domestic cats by vaccination or by therapeutic drug application.

Infection of male rats with Toxoplasma gondii induces effort-aversion in a T-maze decision-making task.

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Tan, Donna; Vyas, Ajai

2016-03-01

Rats chronically infected with protozoan Toxoplasma gondii exhibit greater delay aversion in an inter-temporal task. Moreover T. gondii infection also results in dendritic atrophy of basolateral amygdala neurons. Basolateral amygdala is reported to bias decision making towards greater effortful alternatives. In this context, we report that T. gondii increases effort aversion in infected male rats. This host-parasite association has been widely studied in the context of loss of innate fear in the infected males. It is suggested that reduced fear towards predators reflects a parasitic behavioral manipulation to enhance trophic transmission of T. gondii. Observations reported here extend this paradigm away from a monolithic change in fear and towards a multi-dimensional change in decision making. Copyright © 2016 Elsevier Inc. All rights reserved.

A seroepidemiological survey of Toxoplasma gondii infection in referred dogs to Veterinary Hospital of Ahvaz, Iran

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Fatemeh Zarra-Nezhad

2017-12-01

Full Text Available Toxoplasma gondii is an intracellular protozoan parasite, which is the cause of toxoplasmosis and can infect a wide variety of warm-blooded animals, including dogs and humans. The present study evaluated the seroprevalence of T. gondii infection in pet dogs in Ahvaz, southwest city of Iran, and investigated the related possible risk factors. A total of 180 serum samples were collected from dogs referred to Veterinary Hospital of Ahvaz. The samples were then tested by indirect enzyme-linked immunosorbent assay. The prevalence of T. gondii antibodies were 46.67%. Logistic regression and chi square tests were used for evaluating of risk factors. The positivity increased statistically significantly with dog’s gender (56% females and 39% males, P = 0.001, age (18% in <2 years old, 96% in ≥4 years old, P = 0.001 and place of living (47% outdoor dogs 38% house hold dogs, P = 0.025. However, no statistically significant association was found with dogs’ breed, deworming, food ingestion or contact with cats. Overall, the results showed a relatively high seroprevalence of T. gondii infection in dogs in southwest Iran (Ahvaz and proved association of T. gondii prevalence rates with the dog’s age, gender and place of living. Keywords: Toxoplasma gondii, Toxoplasmosis, Indirect enzyme-linked immunosorbent assay, Outdoor dogs, House hold dogs, Deworming

Toxoplasmosis in dogs: First report of Toxoplasma gondii infection in any animal species in Angola

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Despite the worldwide importance of zoonotic parasite Toxoplasma gondii nothing is known of toxoplasmosis in animals in Angola. The present study aimed at estimating the seroprevalence and also assessing correlates of T. gondii infection in pet dogs from Luanda, Angola. Dogs (n = 103) brought to a v...

Isolation of Viable Toxoplasma gondii from Tissues and Feces of Cats from Addis Ababa, Ethiopia

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Cats are important in the epidemiology of Toxoplasma gondii because they are the only hosts that excrete environmentally resistant oocysts in feces. In the present study, hearts, serum, and feces from 36 feral cats from Addis Ababa area, Ethiopia were examined for T. gondii infection. Antibodies to ...

Fatal toxoplasmosis in an immunosuppressed domestic cat from Brazil caused by Toxoplasma gondii clonal type I

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Hilda Fátima de Jesus Pena

Full Text Available Abstract The objective of the study was to report on a fatal case of feline toxoplasmosis with coinfection with the feline leukemia virus (FeLV. A domestic cat (Felis silvestris catus presented intense dyspnea and died three days later. In the necropsy, the lungs were firm, without collapse and with many white areas; moderate lymphadenomegaly and splenomegaly were also observed. The histopathological examination showed severe necrotic interstitial bronchopneumonia and mild necrotic hepatitis, associated with intralesional cysts and tachyzoites of Toxoplasma gondii that were positive by anti-T. gondii immunohistochemical (IHC evaluation. The bone marrow showed chronic myeloid leukemia and the neoplastic cells were positive by anti-FeLV IHC evaluation. DNA extracted from lungs was positive for T. gondii by PCR targeting REP-529. T. gondii was characterized by PCR-RFLP and by the microsatellites technique. ToxoDB-PCR-RFLP #10, i.e. the archetypal type I, was identified. Microsatellite analysis showed that the strain was a variant of type I with two atypical alleles. This was the first time that a T. gondii clonal type I genotype was correlated with a case of acute toxoplasmosis in a host in Brazil.

Seroprevalence and Risk Factors Associated with Seropositivity to Toxoplasma gondii among Stray and Domestic Cats (Felis silvestris catus

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Christel Bohn T. Garcia

2014-12-01

Full Text Available Toxoplasma gondii is a protozoan parasite that causes toxoplasmosis. It is widespread in the environment and infects a variety of warm-blooded animals, causing miscarriages and birth problems. Previous studies in the Philippines have determined the seropositivity of T. gondii in humans. However, the seroprevalence of the parasite among household pets, par ticularly its feline def initive host, remains insufficient . This study aimed to: (1 determine the seroprevalence of T. gondii antibodies among domestic and stray cats in the Philippines; and, (2 to analyze the risk factors associated with seropositivity. Blood samples from 59 domestic and stray cats were collected and tested for T. gondii seropositivity using a commercially available indirect ELISA kit, while pet owners and handlers were given questionnaires about their cats. Thirteen or 22.03% of the cats were seropositive to T. gondii, and risk factor analysis revealed a significant difference between domestic and stray cats with regard to diet (p = 0.026, OR = 8.333, c = 0.299 and domestication (p = 0.039, OR = 5.000, c = 0.276. Cats fed with table food tested 31.43% seropositive compared to the 4.35% of those fed with cat food, whereas 33.33% of the stray cats were seropositive compared to 7.69% for domestic cats. Odds ratio test showed that the risk factors studied were associated with higher likelihood of T. gondii seropositivity. These results implicate diet and environment in the transmission dynamics of T. gondii among cats.

Incidence of adult brain cancers is higher in countries where the protozoan parasite Toxoplasma gondii is common

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Thomas, Frédéric; Lafferty, Kevin D.; Brodeur, Jacques; Elguero, Eric; Gauthier-Clerc, Michel; Missé, Dorothée

2011-01-01

We explored associations between the common protozoan parasite Toxoplasma gondii and brain cancers in human populations. We predicted that T. gondii could increase the risk of brain cancer because it is a long-lived parasite that encysts in the brain, where it provokes inflammation and inhibits apoptosis. We used a medical geography approach based on the national incidence of brain cancers and seroprevalence of T. gondii. We corrected reports of incidence for national gross domestic product b...

Molecular cloning and characterization of a cDNA encoding ...

African Journals Online (AJOL)

enoh

2012-03-29

Mar 29, 2012 ... Cloning and Functional. Expression of Cycloartenol Synthases from Mangrove Species. Rhizophora stylosa Griff. And Kandelia candel (L.) Druce. Biosci. Biotechnol. Biochem. 71(7): 1788-1792. Felsenstein J (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution. 39:783-791 ...

Isolation and characterization of new genetic types of toxoplasma gondii and prevalence of trichinella murrelli from black bear (Ursus americanus)

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Black bears (Ursus americanus) are hosts for two important zoonotic parasites, Toxoplasma gondii and Trichinella spp. and bears are hunted for human consumption in the USA. Little is known of the genetic diversity of T. gondii circulating in wildlife. In the present study, antibodies to T. gondii we...

High prevalence and genotypes of Toxoplasma gondii isolated from goats, from a retail meat store, destined for human consumption in the USA.

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Dubey, J P; Rajendran, C; Ferreira, L R; Martins, J; Kwok, O C H; Hill, D E; Villena, I; Zhou, H; Su, C; Jones, J L

2011-07-01

Little information is available concerning the presence of viable Toxoplasma gondii in tissues of goats worldwide. In the present study, hearts of 234 goats obtained from a local USA grocery store were examined for T. gondii infection. Blood clot or fluid removed from each heart was tested for antibodies to T. gondii by using the modified agglutination test (MAT). Antibodies to T. gondii were found in 125 (53.4%) of 234 goats, with titers of 1:5 in 20, 1:10 in 44, 1:20 in 16, 1:40 in five, 1:160 in five, 1:320 in five, and 1:640 or higher in 30 goats. Hearts of 112 goats (46 goats goats 1:10 or higher) were used for isolation of viable T. gondii by bioassays in mice. For bioassays, 50 g of the myocardium were digested in an acid pepsin solution and the digest inoculated into mice; the recipient mice were examined for T. gondii infection. Toxoplasma gondii was isolated from 29 goats; from hearts of one of 46 with titers of goat strains. Taken together, these results indicate high parasite prevalence and moderate genetic diversity of T. gondii in goats, which have important implications in public health. We believe this is the first genetic analysis of T. gondii isolates from goats in the USA. Published by Elsevier Ltd.

Seroprevalence and Risk Factors of Toxoplasma gondii Infection in Buffaloes, Sheep and Goats in Yunnan Province, Southwestern China

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Fengcai ZOU

2015-12-01

Full Text Available Background: The seroprevalence of Toxoplasma gondii infection in buffaloes, sheep and goats in Yunnan Province, southwestern China was conducted between May 2012 and December 2013.Methods: A total of 973 (427 buffaloes, 154 sheep and 392 goats serum samples were collected from seven administrative regions of Yunnan Province, and exam­ined for T. gondii antibodies by indirect hemagglutination (IHA test. Some risk fac­tors related to species, age, gender and geographical origin were determined using a multinomial logistic regression.Results: The overall seroprevalence of T. gondii in ruminant species was estimated at 11.9%. The final logistic regression model demonstrated that host species and geographical origin were the main risk factors associated with T. gondii infection (P﹤0.05.Conclusion: Taken together, the results of the present study revealed a high expo­sure to T. gondii in ruminant species in Yunnan Province, which has an important implication for public health.

Toxoplasma gondii: 1908-2008, homage to Nicolle, Manceaux and Splendore

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David J P Ferguson

2009-03-01

Full Text Available The discovery of Toxoplasma gondii independently by Nicolle and Manceaux (1908 and Splendore (1908 was to open a "Pandora's Box" that has led research on this parasite into a number of scientific disciplines. In the 100 years since its discovery, the mystery surrounding T. gondii and its inter-relationship with humans has continued to provide a stimulating source of material in many areas of research, resulting in the publication of almost 20,000 papers and a number of books. This flood of diverse information shows no sign of abating, with an average of 10 papers per week appearing in PubMed. Herein, it is impossible to do more than provide a very superficial comment on what has become a massive body of scientific information. T. gondii has many unique features and seems to be the "exception to almost every rule" thus acting as a focus for research in disciplines from epidemiology to immunology to human behaviour to cell biology to human disease. In this review a number of the historical advances will be mentioned and combined with a description of the basic biology of the parasite.

Construction of recombinant DNA clone for bovine viral diarrhea virus

International Nuclear Information System (INIS)

Yeo, S.G.; Cho, H.J.; Masri, S.A.

1992-01-01

Molecular cloning was carried out on the Danish strain of bovine viral diarrhea virus (BVDV) to construct strategy for the diagnostic tools and effective vaccine of BVD afterwards. A recombinant DNA clone (No. 29) was established successfully from cDNA for viral RNA tailed with adenine homopolymer at 3 -end. 32 P-labeled DNA probes of 300~1, 800bp fragments, originating from the clone 29, directed specific DNA-RNA hybridization results with BVDV RNA. Recombinant DNA of the clone 29 was about 5,200bp representing 41.6% of the full length of Danish strain's RNA, and restriction sites were recognized for EooR I, Sst I, Hind III and Pst I restriction enzymes in the DNA fragment

Toxoplasma gondii Infection and Mixed Anxiety and Depressive Disorder: A Case-Control Seroprevalence Study in Durango, Mexico.

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Alvarado-Esquivel, Cosme; Sanchez-Anguiano, Luis Francisco; Hernandez-Tinoco, Jesus; Berumen-Segovia, Luis Omar; Torres-Prieto, Yazmin Elizabeth; Estrada-Martinez, Sergio; Perez-Alamos, Alma Rosa; Ortiz-Jurado, Maria Nalleli; Molotla-de-Leon, Gabriel; Beristain Garcia, Isabel; Rabago-Sanchez, Elizabeth; Liesenfeld, Oliver

2016-07-01

The parasite Toxoplasma gondii (T. gondii) may invade the brain and might induce behavioral changes. We sought to determine the association of T. gondii infection and mixed anxiety and depressive disorder. Through an age- and gender-matched case-control seroprevalence study, we examined 65 patients suffering from mixed anxiety and depressive disorder (WHO ICD-10 code: F41.2) attending in a public hospital of mental health and 260 control subjects without this disorder from the general population. Sera of participants were analyzed for anti-Toxoplasma IgG and IgM antibodies using enzyme-linked immunoassays. Fifteen (23.1%) of the 65 patients and 18 (6.9%) of the 260 controls had anti-T. gondii IgG antibodies (odds ratio (OR): 4.03; 95% confidence interval (CI): 1.90 - 8.53; P 150 IU/mL) anti-T. gondii IgG levels was similar in cases and controls (OR: 0.25; 95% CI: 0.05 - 1.06; P = 0.05). Seroprevalence was similar in male cases and controls (P = 1.0); however, seroprevalence was significantly higher in female cases than in female controls (OR: 7.08; 95% CI: 2.83 - 17.67; P mixed anxiety and depressive disorder. Further research to confirm this association and to determine the seroepidemiology of T. gondii infection in patients with this disorder is needed.

Antibody Prevalence and Isolation of Viable Toxoplasma gondii from Raptors in the Southeastern USA.

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Love, David; Kwok, Oliver C; Verma, Shiv Kumar; Dubey, Jitender P; Bellah, Jamie

2016-07-01

Raptors are good indicators of the prevalence of Toxoplasma gondii in the environment because they prey on small mammals and birds. These prey species are a major source of infection in domestic cats ( Felis catus ), which shed the environmentally resistant oocysts. We assessed T. gondii infection in 281 opportunistically available raptors at a rehabilitation facility between 2012 and 2014. Antibodies to T. gondii were assayed by a modified agglutination test (cutoff 1:25) and found in serum of 22/71 Red-tailed Hawks ( Buteo jamaicensis ), 25/54 Barred Owls ( Strix varia ), 9/41 Red-shouldered Hawks ( Buteo lineatus ), 13/28 Great Horned Owls ( Bubo virginianus ), 6/20 Broad-winged Hawks ( Buteo platypterus ), 2/16 Eastern Screech Owls (Megascops asio), 12/13 Bald Eagles ( Haliaeetus leucocephalus ), 6/12 Cooper's Hawks ( Accipiter cooperii ), 1/8 Black Vultures ( Coragyps atratus ), and 1/1 Golden Eagle ( Aquila chrysaetos ). Antibodies were not detected in 5 Barn Owls ( Tyto alba ), 3 American Kestrels ( Falco sparverius ), 1 Mississippi Kite ( Ictinia mississippiensis ), and 1 Osprey ( Pandion haliaetus ). Viable T. gondii was isolated from the tissues of 1 antibody-positive Barred Owl and identified as a strain having type II alleles at all 10 loci tested, except one (ToxoDB polymerase chain reaction-restriction fragment length polymorphism genotype 3). Type II strain is the most common strain in the US. Results of this study indicate a high prevalence of T. gondii in some raptor species and the first reported genotyping from a Barred Owl.

Structure-Based Analysis of Toxoplasma gondii Profilin: A Parasite-Specific Motif Is Required for Recognition by Toll-Like Receptor 11

Energy Technology Data Exchange (ETDEWEB)

K Kucera; A Koblansky; L Saunders; K Frederick; E De La Cruz; S Ghosh; Y Modis

2011-12-31

Profilins promote actin polymerization by exchanging ADP for ATP on monomeric actin and delivering ATP-actin to growing filament barbed ends. Apicomplexan protozoa such as Toxoplasma gondii invade host cells using an actin-dependent gliding motility. Toll-like receptor (TLR) 11 generates an innate immune response upon sensing T. gondii profilin (TgPRF). The crystal structure of TgPRF reveals a parasite-specific surface motif consisting of an acidic loop, followed by a long {beta}-hairpin. A series of structure-based profilin mutants show that TLR11 recognition of the acidic loop is responsible for most of the interleukin (IL)-12 secretion response to TgPRF in peritoneal macrophages. Deletion of both the acidic loop and the {beta}-hairpin completely abrogates IL-12 secretion. Insertion of the T. gondii acidic loop and {beta}-hairpin into yeast profilin is sufficient to generate TLR11-dependent signaling. Substitution of the acidic loop in TgPRF with the homologous loop from the apicomplexan parasite Cryptosporidium parvum does not affect TLR11-dependent IL-12 secretion, while substitution with the acidic loop from Plasmodium falciparum results in reduced but significant IL-12 secretion. We conclude that the parasite-specific motif in TgPRF is the key molecular pattern recognized by TLR11. Unlike other profilins, TgPRF slows nucleotide exchange on monomeric rabbit actin and binds rabbit actin weakly. The putative TgPRF actin-binding surface includes the {beta}-hairpin and diverges widely from the actin-binding surfaces of vertebrate profilins.

Comparison of three diagnostic methods for the detection of Toxoplasma gondii in free range chickens.

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Hamidinejat, H; Nabavi, L; Mayahi, M; Ghourbanpoor, M; Pourmehdi Borojeni, M; Norollahi Fard, S; Shokrollahi, M

2014-09-01

Detection of Toxoplasma gondii in free range chickens is an indicator of the prevalence and distribution pattern of T. gondii in the environment. For this purpose, serologic assays especially modified agglutination test (MAT) is the main approach in the literature. The main goal of this study was to compare the polymerase chain reaction (PCR) based on amplification of first internal transcribed spacer (ITS-1) of ribosomal DNA gene, ELISA, and MAT to demonstrate T. gondii infection in free range chicken. A total of 106 adult free - range chickens were killed. Blood, whole heart and brain samples were taken. Sera were examined for the presence of T. gondii antibodies by ELISA and MAT as well. Selected tissues were used for PCR and bioassay in mice. The results revealed that 48.11%, 51.89%, 46.23% and 27.36% of chickens were positive in ELISA, MAT, PCR and bioassay in mice respectively. Good correlation between the results of PCR, ELISA and MAT were detected, but not with bioassay in mice. Compared with PCR, the sensitivity and specificity of ELISA were 92.16% and 96.36% respectively and also for MAT, the sensitivity was 81.81% and the specificity was 92.15%. The specific diagnosis of T. gondii infection in chickens is central to a better understanding of the epidemiology and dynamics of transmission among the various host population and is particularly important for planning effective optimal prevention and control programs. Our data in the present study demonstrated that PCR, ELISA and the MAT are helpful and precise methods to detect T. gondii in naturally infected free-range chickens.

Toxoplasma gondii and pre-treatment protocols for polymerase chain reaction analysis of milk samples: a field trial in sheep from Southern Italy

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Alice Vismarra

2017-02-01

Full Text Available Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. Ingestion of raw milk has been suggested as a risk for transmission to humans. Here the authors evaluated pre-treatment protocols for DNA extraction on T. gondii tachyzoite-spiked sheep milk with the aim of identifying the method that resulted in the most rapid and reliable polymerase chain reaction (PCR positivity. This protocol was then used to analyse milk samples from sheep of three different farms in Southern Italy, including real time PCR for DNA quantification and PCR-restriction fragment length polymorphism for genotyping. The pre-treatment protocol using ethylenediaminetetraacetic acid and Tris-HCl to remove casein gave the best results in the least amount of time compared to the others on spiked milk samples. One sample of 21 collected from sheep farms was positive on one-step PCR, real time PCR and resulted in a Type I genotype at one locus (SAG3. Milk usually contains a low number of tachyzoites and this could be a limiting factor for molecular identification. Our preliminary data has evaluated a rapid, cost-effective and sensitive protocol to treat milk before DNA extraction. The results of the present study also confirm the possibility of T. gondii transmission through consumption of raw milk and its unpasteurised derivatives.

Pulmonary exposure to single-walled carbon nanotubes does not affect the early immune response against Toxoplasma gondii

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Swedin Linda

2012-05-01

Full Text Available Abstract Background Single-walled carbon nanotubes (SWCNT trigger pronounced inflammation and fibrosis in the lungs of mice following administration via pharyngeal aspiration or inhalation. Human exposure to SWCNT in an occupational setting may occur in conjunction with infections and this could yield enhanced or suppressed responses to the offending agent. Here, we studied whether the sequential exposure to SWCNT via pharyngeal aspiration and infection of mice with the ubiquitous intracellular parasite Toxoplasma gondii would impact on the immune response of the host against the parasite. Methods C57BL/6 mice were pre-exposed by pharyngeal administration of SWCNT (80 + 80 μg/mouse for two consecutive days followed by intravenous injection with either 1x103 or 1x104 green fluorescence protein and luciferase-expressing T. gondii tachyzoites. The dissemination of T. gondii was monitored by in vivo bioluminescence imaging in real time for 7 days and by plaque formation. The inflammatory response was analysed in bronchoalveolar lavage (BAL fluid, and by assessment of morphological changes and immune responses in lung and spleen. Results There were no differences in parasite distribution between mice only inoculated with T. gondii or those mice pre-exposed for 2 days to SWCNT before parasite inoculum. Lung and spleen histology and inflammation markers in BAL fluid reflected the effects of SWCNT exposure and T. gondii injection, respectively. We also noted that CD11c positive dendritic cells but not F4/80 positive macrophages retained SWCNT in the lungs 9 days after pharyngeal aspiration. However, co-localization of T. gondii with CD11c or F4/80 positive cells could not be observed in lungs or spleen. Pre-exposure to SWCNT did not affect the splenocyte response to T. gondii. Conclusions Taken together, our data indicate that pre-exposure to SWCNT does not enhance or suppress the early immune response to T. gondii in mice.

Seroprevalence of Toxoplasma gondii in free-ranging and feral cats on Amami Oshima Island, Japan.

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Matsuu, Aya; Yokota, Shin-Ichi; Ito, Keiko; Masatani, Tatsunori

2017-11-17

On Amami Oshima Island, free-ranging and feral cats are harmful to wildlife populations. In this study, the seroprevalence of Toxoplasma gondii in these cats was examined using a newly developed Gaussia luciferase immunoprecipitation system assay. Of 1,363 cats, 123 cats (9.0%) was positive for T. gondii. The prevalence was significantly different in different areas; among cats in the rural area, where many wild animals live, including endangered species, T. gondii infection was more prevalent than in the urban area of the island. This finding indicates a possible risk to wildlife of infection from free-ranging and feral cats. Therefore, management of cats is important for wildlife conservation.

Spread of carbapenem-resistant international clones of Acinetobacter baumannii in Turkey and Azerbaijan: a collaborative study

NARCIS (Netherlands)

Ahmed, S.S.; Alp, E.; Ulu-Kilic, A.; Dinc, G.; Aktas, Z.; Ada, B.; Bagirova, F.; Baran, I.; Ersoy, Y.; Esen, S.; Guven, T.G.; Hopman, J.; Hosoglu, S.; Koksal, F.; Parlak, E.; Yalcin, A.N.; Yilmaz, G.; Voss, A.; Melchers, W.J.

2016-01-01

Epidemic clones of Acinetobacter baumannii, described as European clones I, II, and III, are associated with hospital epidemics throughout the world. We aimed to determine the molecular characteristics and genetic diversity between European clones I, II, and III from Turkey and Azerbaijan. In this

Design of a Risk based Control System for Toxoplasma gondii in a pork supply chain

NARCIS (Netherlands)

Heres, L.; Swanenburg, M.; Koeijer, de A.A.; Boender, G.J.; Oorburg, D.; Urlings, B.; Wisselink, H.J.

2015-01-01

Toxoplasma gondii is recognized as one of the major foodborne pathogens. The European Food Safety Authority (EFSA) concluded that T. gondii is one of the public health hazards in pigs to be covered within a modern meat inspection and advised to include serological testing of pigs and audits of pig

Behavioral changes in Rattus norvegicus coinfected by Toxocara canis and Toxoplasma gondii

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Maisa Leite de Queiroz

2013-02-01

Full Text Available Using an elevated plus maze apparatus and an activity cage, behavioral changes in Rattus norvegicus concomitantly infected by Toxocara canis and Toxoplasma gondii were studied, during a period of 120 days. Rats infected by Toxocara canis or Toxoplasma gondii showed significant behavioral changes; however, in the group coinfected by both parasites a behavioral pattern similar to that found in the group not infected was observed thirty days after infection, suggesting the occurrence of modulation in the behavioral response.

High seroprevalence of Toxoplasma gondii infection in a subset of Mexican patients with work accidents and low socioeconomic status.

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Alvarado-Esquivel, Cosme; Torres-Castorena, Alejandro; Liesenfeld, Oliver; Estrada-Martínez, Sergio; Urbina-Álvarez, Jesús D

2012-01-11

Toxoplasma gondii has been associated with reflex impairment and traffic accidents. It is unknown whether Toxoplasma infection might be associated with work accidents. Therefore, using a case-control seroprevalence study design, 133 patients with a recent work accident and 266 control subjects of the general population from the same region were examined with enzyme-linked immunoassays for the presence and levels of anti-Toxoplasma IgG antibodies and anti-Toxoplasma IgM antibodies. Socio-demographic, work, clinical and behavioral characteristics from each worker were obtained. Eleven (8.3%) of 133 patients, and 14 (5.3%) of 266 controls had anti-T. gondii IgG antibodies. Anti-T. gondii IgG levels were higher than 150 IU/ml in 8 (6%) patients and 10 (3.8%) controls. Anti-T. gondii IgM antibodies were found in one (0.8%) of the workers, and in 6 (2.3%) of the controls. No statistically significant differences in the IgG seroprevalences, frequencies of high IgG levels, and IgM seroprevalences among patients and controls were found. In contrast, a low socio-economic level in patients with work accidents was associated with Toxoplasma seropositivity (P = 0.01). Patients with work accidents and low socioeconomic status showed a significantly (OR = 3.38; 95% CI: 0.84-16.06; P = 0.04) higher seroprevalence of T. gondii infection than controls of the same socioeconomic status (15.1% vs. 5%, respectively). Multivariate analysis showed a positive association of T. gondii infection with boar meat consumption (OR = 3.04; 95% CI: 1.03-8.94; P = 0.04). In contrast, a negative association between T. gondii infection and national trips (OR = 0.40; 95% CI: 0.17-0.96; P = 0.04), sausage consumption (OR = 0.20; 95% CI: 0.05-0.68; P = 0.01), and ham consumption (OR = 0.16; 95% CI: 0.05-0.51; P = 0.002) was found. In the study described here seropositivity to T. gondii was associated to work accidents in a subset of patients with low socioeconomic status. This is the first report of an

Seroprevalence of Toxoplasma gondii and Neospora caninum ...

African Journals Online (AJOL)

The aim of this study was to compare the seroprevalence of Toxoplasma gondii and Neospora caninum in goats from two Argentinean provinces raised under different management conditions. A total of 2922 serum samples from adult goats of Córdoba (n=2187) and Buenos Aires provinces (n= 735), Argentina, were ...

Risk factors related to Toxoplasma gondii seroprevalence in indoor-housed Dutch dairy goats.

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Deng, Huifang; Dam-Deisz, Cecile; Luttikholt, Saskia; Maas, Miriam; Nielen, Mirjam; Swart, Arno; Vellema, Piet; van der Giessen, Joke; Opsteegh, Marieke

2016-02-01

Toxoplasma gondii can cause disease in goats, but also has impact on human health through food-borne transmission. Our aims were to determine the seroprevalence of T. gondii infection in indoor-housed Dutch dairy goats and to identify the risk factors related to T. gondii seroprevalence. Fifty-two out of ninety approached farmers with indoor-kept goats (58%) participated by answering a standardized questionnaire and contributing 32 goat blood samples each. Serum samples were tested for T. gondii SAG1 antibodies by ELISA and results showed that the frequency distribution of the log10-transformed OD-values fitted well with a binary mixture of a shifted gamma and a shifted reflected gamma distribution. The overall animal seroprevalence was 13.3% (95% CI: 11.7–14.9%), and at least one seropositive animal was found on 61.5% (95% CI: 48.3–74.7%) of the farms. To evaluate potential risk factors on herd level, three modeling strategies (Poisson, negative binomial and zero-inflated) were compared. The negative binomial model fitted the data best with the number of cats (1–4 cats: IR: 2.6, 95% CI: 1.1–6.5; > = 5 cats:IR: 14.2, 95% CI: 3.9–51.1) and mean animal age (IR: 1.5, 95% CI: 1.1–2.1) related to herd positivity. In conclusion, the ELISA test was 100% sensitive and specific based on binary mixture analysis. T. gondii infection is prevalent in indoor housed Dutch dairy goats but at a lower overall animal level seroprevalence than outdoor farmed goats in other European countries, and cat exposure is an important risk factor.

Toxoplasma gondii antibodies in domiciled cats from rio branco Municipality, Acre State, Brazil

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Soraia Figueiredo de Souza

2015-12-01

Full Text Available Blood samples were collected from 89 cats to assess the prevalence of IgG antibodies against Toxoplasma gondii by indirect immunofluorescence (IIF and the possible risk factors associated with feline Toxoplasma gondii infection. An epidemiological questionnaire was developed and implemented for owners of domestic cats domiciled in Rio Branco, Acre. The results were statistically evaluated with the odds ratio and chi-square tests, considering the significance level of 5%. Of 89 animals’ samples, 22 had antibodies against Toxoplasma gondii. Among the 22 reactive animals, 15 (68.19% were female, 15 (68.19% were less than one year old and 20 (90% were cross breed. Concerning risk factors, there was no difference (p > 0.05 between the variables evaluated by the chi-square test. Moreover, 16 (72% cats were fed a mixed diet, 20 (90% of the cats had hunting habits, 18 (81% had contact with animals of another species, 11 (50% had access to the street, and 22 (95% lived in homes that had areas of grass or dirt. In conclusion, the prevalence of anti-Toxoplasma gondii in domestic cats was 22.7%, and there were no significant risk factors for feline toxoplasmosis in the municipality of Rio Branco, Acre.

[Awareness and infection of Toxoplasma gondii in married childbearing women in Chengde Region].

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Li, Xue-jing; Xu, Tao; Song, Ren-hao

2014-08-01

To understand Toxoplasma gondii infection and awareness condition of married childbearing women in Chengde Region, so as to provide the evidence for the establishment of control measures. Totally 733 married childbearing women who took physical examination in Chengde Hospital of Traditional Chinese Medicine from July to December in 2013 were investigated by questionnaire to understand the awareness condition on T. gondii infection, then 490 women among them from 3 counties and 2 districts were randomly chosen to detect the Toxoplasma antibodies by ELISA. A total of 733 questionnaires were returned, and 126 women knew related knowledge about T. gondii infection, and the awareness rate was 17.19%( 126/733). Sixty-three women were determined as infected cases, and the infection rate was 12.86%( 63/490). The infection rates of the women who with higher educational level, working as medical staff and living in urban were lower, and the awareness rates of them were higher. The infection rate of T. gondii among the married childbearing women in Chengde Region is high, and the awareness rate of them is low. In order to decrease the infection rate as well as to increase the awareness rate of the population, the health education should be strengthened.

Brain cystogenesis capacity of Toxoplasma gondii, avirulent Tehran strain in mice

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Mehrzad Saraei

2014-09-01

Full Text Available Objective: To investigate the brain cystogenesis capacity of Tehran strain of Toxoplasma gondii (T. gondii that had been isolated from a patient with lymphadenitis in 1973. Methods: A volume of 0.5 mL mice brain suspension containing 20 tissue cysts of Tehran strain of T. gondii was inoculated intraperitoneally to each of 25 male BALB/c mice. The number of brain cysts was counted in unstained squash-smears for 10 mice during weeks 7-9 and for 15 mice during weeks 13-14 post-infection. Nonparametric test of Mann-Whitney was used to demonstrate means differences. Results: There was a significant difference in the means for the number of brain cysts between weeks 7-9 (228.3依144.8 and weeks 13-14 (1 239.8依429.3 post-infection (P<0.05. The minimum and the maximum of cysts were 70 and 1 531 during weeks 7-9 post-infection, and 12 and 5 170 during weeks 13-14 post-infection, respectively. The mean number of brain cysts in the right cerebral hemisphere was insignificantly higher than that of the left cerebral hemisphere. Furthermore, the number of cysts counted in the right or the left hemispheres was significantly higher than those enumerated for cerebellum+brain stem altogether. Conclusions: It is concluded that the brain cystogenesis capacity of T. gondii, Tehran strain shows enormous variation in mice regarding the duration of infection. In addition, the cystogenesis observed in cerebellum+brain stem is lower than the right and left cerebral hemispheres.

Serosurvey of Leptospira spp. and Toxoplasma gondii in rats captured from two zoos in Southern Brazil.

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Pellizzaro, Maysa; Conrado, Francisco de Oliveira; Martins, Camila Marinelli; Joaquim, Sâmea Fernandes; Ferreira, Fernando; Langoni, Helio; Biondo, Alexander Welker

2017-01-01

Norway rats (Rattus norvegicus) are zoonotic reservoirs for Leptospira spp. and Toxoplasma gondii, and influence diseases in urban areas. Free-ranging and laboratory-raised rats from two zoos in southern Brazil were tested for Leptospira spp. and T. gondii using microscopic agglutination and modified agglutination tests, respectively. Overall, 25.6% and 4.6% free-ranging rats tested positive for Leptospira spp. and T. gondii, respectively, with co-seropositivity occurring in two animals. For laboratory-raised rats, 20% tested positive for Leptospira spp. Also, Leptospira biflexa serovar Patoc and Leptospira noguchii serovar Panama were found. Serosurveys can show the environmental prevalence of zoonotic pathogens.

Host Mitochondrial Association Evolved in the Human Parasite Toxoplasma gondii via Neofunctionalization of a Gene Duplicate.

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Adomako-Ankomah, Yaw; English, Elizabeth D; Danielson, Jeffrey J; Pernas, Lena F; Parker, Michelle L; Boulanger, Martin J; Dubey, Jitender P; Boyle, Jon P

2016-05-01

In Toxoplasma gondii, an intracellular parasite of humans and other animals, host mitochondrial association (HMA) is driven by a gene family that encodes multiple mitochondrial association factor 1 (MAF1) proteins. However, the importance of MAF1 gene duplication in the evolution of HMA is not understood, nor is the impact of HMA on parasite biology. Here we used within- and between-species comparative analysis to determine that the MAF1 locus is duplicated in T. gondii and its nearest extant relative Hammondia hammondi, but not another close relative, Neospora caninum Using cross-species complementation, we determined that the MAF1 locus harbors multiple distinct paralogs that differ in their ability to mediate HMA, and that only T. gondii and H. hammondi harbor HMA(+) paralogs. Additionally, we found that exogenous expression of an HMA(+) paralog in T. gondii strains that do not normally exhibit HMA provides a competitive advantage over their wild-type counterparts during a mouse infection. These data indicate that HMA likely evolved by neofunctionalization of a duplicate MAF1 copy in the common ancestor of T. gondii and H. hammondi, and that the neofunctionalized gene duplicate is selectively advantageous. Copyright © 2016 by the Genetics Society of America.

Anti-N-methyl-D-aspartate receptor encephalitis associated with acute Toxoplasma gondii infection: A case report.

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Cai, Xiaotang; Zhou, Hui; Xie, Yongmei; Yu, Dan; Wang, Zhiling; Ren, Haitao

2018-02-01

Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis has been recognized as the most frequent autoimmune encephalitis in children. Several infectious agents have been implicated in anti-NMDA encephalitis. A previously healthy immunocompetent 9-year-old girl first presented with seizures, headaches and vomiting. Cerebrospinal fluid and brain magnetic resonance imaging were normal. After one week onset, the patient gradually developed unexplained personality and behavior changes, accompanied by fever and seizures again. Repeated CSF analysis revealed a slightly lymphocytic predominant pleocytosis and positive anti-NMDAR antibody. A variety of pathogenic examinations were negative, except for positive toxoplasma IgM and IgG. The patient was diagnoses for anti-NMDA encephalitis associated with acute acquired toxoplasma gondii infection. The patient received 10 days azithromycin for treatment of acquired toxoplasma infection. The parents refuse immunotherapy because substantial recovery from clinical symptoms. The patient was substantially recovered with residual mild agitation after therapy for acquired toxoplasma gondii infection. Two months later, the patient was completely devoid of symptoms, and the levels of serum IgM and IgG of toxoplasma gondii were decreased. Acquired toxoplasma gondii infection may trigger anti-NMDAR encephalitis in children, which has not been reported previously. Clinicians should assess the possibility of toxoplasma gondii infection when evaluating a patient with anti-NMDA encephalitis.

Seropositivity of Toxoplasma gondii and Toxocara spp. in Children with Cryptogenic Epilepsy, Benha, Egypt.

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Eraky, Maysa Ahmad; Abdel-Hady, Soha; Abdallah, Karim Fetouh

2016-06-01

The present study aimed to investigate the possible association of Toxoplasma gondii and Toxocara spp. infections with cryptogenic epilepsy in children. The study was carried out between June 2014 and March 2015. Total 90 children (40 with cryptogenic epilepsy, 30 with non-cryptogenic epilepsy, and 20 healthy control children) were evaluated to determine the anti-Toxocara and anti-T. gondii IgG seropositivity using ELISA kits. Epileptic cases were selected from those attending the pediatrics outpatient clinic of Benha University Hospital, Pediatrics Neurology Unit, and from Benha Specialized Hospital of children. The results showed that the level of anti-T. gondii IgG seropositivity was significantly higher among children with cryptogenic epilepsy (20%) than among children with non-cryptogenic children (0%). In healthy controls (10%), there was no association between toxocariasis seropositivity and cryptogenic epilepsy (only 5.7%; 4 out of 70 cases) among cases and 10% (2 out of 20) among controls. Among toxocariasis IgG positive cases, 3 (7.5%) were cryptogenic, and only 1 (3.3%) was non-cryptogenic. These statistically significant results support the association between T. gondii infection and cryptogenic epilepsy while deny this association with toxocariasis.

Prevalence of Toxoplasma gondii infections in Pennsylvania black bears, Ursus americanus.

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Briscoe, N; Humphreys, J G; Dubey, J P

1993-10-01

Serum samples from 665 hunter-killed black bears killed in 1989 to 1992 throughout Pennsylvania (USA) were tested for Toxoplasma gondii antibodies by the agglutination test in dilutions of 1:25, 1:50, and 1:500. Toxoplasma gondii antibodies were found in 535 of 665 (80%) bears. Considering the highest dilutions at which antibodies were detected, prevalences were 10% at 1:25, 37% at 1:50 and 33% at 1:500. No significant difference in antibody prevalence was found between males (79%) and females (80%), but a significant difference was found between juvenile (65%) and adult (83%) bears.

Cloning

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Cloning describes the processes used to create an exact genetic replica of another cell, tissue or organism. ... named Dolly. There are three different types of cloning: Gene cloning, which creates copies of genes or ...

Seroprevalence and risk factors of Neospora spp. and Toxoplasma gondii infections among horses and donkeys in Nigeria, West Africa.

Science.gov (United States)

Bártová, Eva; Sedlák, Kamil; Kobédová, Kateřina; Budíková, Marie; Joel Atuman, Yakubu; Kamani, Joshua

2017-09-26

Neospora spp. and Toxoplasma gondii are considered to be a globally distributed parasites affecting wide range of warm-blooded animals. Neosporosis has caused clinical illness in horses and consumption of horse meat has been epidemiologically linked to clinical toxoplasmosis in humans. This study was conducted to determine Neospora spp. and T. gondii antibodies and risk factors of infection in horses and donkeys from three states of Nigeria. A total of 144 samples were collected from clinically healthy animals (120 horses and 24 donkeys). The sera were tested for antibodies to Neospora spp. and T. gondii by indirect fluorescence antibody test, a titer ≥ 50 was considered positive. Seroprevalence data were statistically analyzed, considering the variables of gender, age, use, state, origin of breed and type of management. Antibodies to Neospora spp. and T. gondii were detected in 8% horses with titers 50 and in 24% horses with titers 50-800, respectively. Co-infection of both parasites was proved in three horses (3%). Statistical differences were found only for T. gondii seroprevalence in horses with different use, locality, origin and management (p-value ≤ 0.05). Antibodies to T. gondii were detected in four (17%) of 24 donkeys with statistical difference (p-value ≤ 0.05) in animals of different use; antibodies to Neospora spp. were not proved in any of the donkeys. This is the first seroprevalence study of Neospora spp. and T. gondii in equids from Nigeria.

What is Cloning?

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Donate Home Cloning What is Cloning What is Cloning Clones are organisms that are exact genetic copies. ... clones made through modern cloning technologies. How Is Cloning Done? Many people first heard of cloning when ...

Enrofloxacin is able to control Toxoplasma gondii infection in both in vitro and in vivo experimental models.

Science.gov (United States)

Barbosa, Bellisa Freitas; Gomes, Angelica Oliveira; Ferro, Eloisa Amália Vieira; Napolitano, Danielle Reis; Mineo, José Roberto; Silva, Neide Maria

2012-06-08

Currently, toxoplasmosis is treated with sulfadiazine and pyrimethamine. However, this treatment presents several adverse side effects; thus, there is a critical need for the development and evaluation of new drugs, which do not present the same problems of the standard therapy. Enrofloxacin is a fluoroquinolone antibiotic known to control infection against several bacteria in veterinary medicine. Recently, this drug has demonstrated protective effects against protozoan parasites such as Neospora caninum. The present study aimed to determine the effect of enrofloxacin in the control of Toxoplasma gondii infection. For this purpose, human foreskin fibroblast (HFF) cells were infected with T. gondii RH strain and treated with sulfadiazine, penicillin/streptomycin, pyrimethamine, or enrofloxacin. Following treatment, we analyzed the infection index, parasite intracellular proliferation and the number of plaques. Additionally, tissue parasitism and histological changes were investigated in the brain of Calomys callosus that were infected with T. gondii (ME49 strain) and treated with either sulfadiazine or enrofloxacin. Enrofloxacin was able to reduce the infection index, intracellular proliferation and the number of plaques in HFF cells infected by T. gondii in comparison with untreated or penicillin/streptomycin-treated ones. Enrofloxacin was more protective against T. gondii in HFF infected cells than sulfadiazine treatment (Penrofloxacin or the associations of sulfadiazine plus pyrimethamine, enrofloxacin plus sulfadiazine or enrofloxacin plus pyrimethamine-treatments were able to reduce the plaque numbers in HFF cells infected by T. gondii when compared to medium, penicillin/streptomycin or sulfadiazine alone. In vivo experiments demonstrated that enrofloxacin diminished significantly the tissue parasitism as well as the inflammatory alterations in the brain of C. callosus infected with T. gondii when compared with untreated animals. Based on our findings, it can

The neurotropic parasite Toxoplasma gondii increases dopamine metabolism

Science.gov (United States)

The common parasite Toxoplasma gondii induces behavioral alterations in its hosts including phenotypes increasing the likelihood of its transmission in rodents and reports of psychobehavioral alterations in humans. We have found that elevated levels of dopamine are associated with the encysted stage...

Age-related Toxoplasma gondii seroprevalence in Dutch wild boar inconsistent with lifelong persistence of antibodies.

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Marieke Opsteegh

Full Text Available Toxoplasma gondii is an important zoonotic pathogen that is best known as a cause of abortion or abnormalities in the newborn after primary infection during pregnancy. Our aim was to determine the prevalence of T. gondii in wild boar to investigate the possible role of their meat in human infection and to get an indication of the environmental contamination with T. gondii. The presence of anti-T. gondii antibodies was determined by in-house ELISA in 509 wild boar shot in 2002/2003 and 464 wild boar shot in 2007. Most of the boar originated from the "Roerstreek" (n = 673 or the "Veluwe" (n = 241. A binormal mixture model was fitted to the log-transformed optical density values for wild boar up to 20 months old to estimate the optimal cut-off value (-0.685 and accompanying sensitivity (90.6% and specificity (93.6%. The overall seroprevalence was estimated at 24.4% (95% CI: 21.1-27.7%. The prevalence did not show variation between sampling years or regions, indicating a stable and homogeneous infection pressure from the environment. The relation between age and seroprevalence was studied in two stages. Firstly, seroprevalence by age group was determined by fitting the binary mixture model to 200 animals per age category. The prevalence showed a steep increase until approximately 10 months of age but stabilized at approximately 35% thereafter. Secondly, we fitted the age-dependent seroprevalence data to several SIR-type models, with seropositives as infected (I and seronegatives as either susceptible (S or resistant (R. A model with a recovery rate (SIS was superior to a model without a recovery rate (SI. This finding is not consistent with the traditional view of lifelong persistence of T. gondii infections. The high seroprevalence suggests that eating undercooked wild boar meat may pose a risk of infection with T. gondii.

Molecular cloning and functional characterization of avian interleukin-19

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The present study describes the cloning and functional characterization of avian interleukin (IL)-19, a cytokine that, in mammals, alters the balance of Th1 and Th2 cells in favor of the Th2 phenotype. The full-length avian IL-19 gene, located on chromosome 26, was amplified from LPS-stimulated chi...

Seroprevalence of toxoplasma gondii infection among patients admitted to al-zahra hospital, isfahan, iran

International Nuclear Information System (INIS)

Mohaghegh, M.A.; Yazdani, H.; Hadipour, M.; Namdar, F.

2015-01-01

Toxoplasma gondii (T. gondii) infection is one of the most common parasitic infections among humans and other warm-blooded animals worldwide. The aim of this study was to evaluate toxoplasmosis status in patients admitted to Al-Zahra hospital, Isfahan, Iran. Methods: This cross-sectional study was conducted from October 2012 to January 2015. During this period, 716 patients referred to Al-Zahra hospital in Isfahan city, Iran, were studied to investigate the IgG and IgM antibodies against T. gondii using ELISA kit. The data were analysed by Chi-square and Fishers exact tests. In addition, the relation of data with age and sex were also examined. Results: Among 716 patients, 21 patients (2.9%) had positive IgM and 288 patients (40.2%) had positive IgG titer against T. gondii. Data analysis by Chi-square and Fishers exact tests revealed that there was no significant relationship between IgG titer and age (p>0.05). Additionally, there was no relationship between IgM titer and age (p>0.05). The data showed that there was no relationship between IgG and IgM antibody titer and sex (p>0.05). Conclusion: The prevalence of toxoplasmosis in Isfahan inhabitants seems fairly high but it can be concluded that the rate of seropositive patient is moderate comparing to other regions of country. Accordingly, the authors propose that all sensitive patients have to be tested for T. gondii antibody in order to prevent the consequences of disease. (author)

Occurrence of antibodies to Toxoplasma gondii in rheas (Rhea americana and ostriches (Struthio camelus from farms of different Brazilian regions

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Amanda Brentano Almeida

Full Text Available This study aimed to verify the occurrence of antibodies againstToxoplasma gondii in rheas (Rhea americana and ostriches (Struthio camelus commercially breeding in Brazil. Blood samples from 20 rheas and 46 ostriches (young and adults were serologically tested using a technique known as modified agglutination test (MAT at an initial titration of 1:16 for ostriches and 1:25 for rheas. Antibodies against T. gondii were found in 50% (10/20 of the rheas, with titers ranging from 1:25 to 1:6,400. The incidence of antibodies against T. gondii in ostriches was 17.4% (8/46 with titers ranging from 1:16 to 1:256. Birds showing titers higher than 1:200 forT. gondii were mainly the young ones. Therefore, rheas and ostriches may be parasitized by T. gondii, showing high levels of antibodies against this parasite.

Seroprevalence of Toxoplasma gondii in free-living European mouflon (Ovis orientalis musimon hunted in central Germany

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Heddergott Mike

2018-01-01

Full Text Available Despite increasing consumption of mouflon (Ovis orientalis musimon meat in Germany, there is currently no surveillance of Toxoplasma gondii infection in populations of these animals and generally little knowledge about the prevalence of this protozoan in German wild ungulates. Between 2011 and 2015, we collected 138 blood samples from a free-living mouflon population in central German and tested sera for the presence of T. gondii antibodies using a modified agglutination test (MAT, cut-off 1:20. Antibodies were detected in 31 of the 138 samples (22.46%. There was a significant difference in seroprevalence between the different age classes, with antibodies to T. gondii more frequent in adults. In contrast, there was no significant difference in seroprevalence depending on sex and year of sample collection. Game meat is frequently consumed as raw or undercooked meat and may therefore represent a potential source of human infection with T. gondii.

Prevalence of Toxoplasma gondii in native donkeys in Mosul

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Kh. J. Hussain

2011-01-01

Full Text Available The aim of the study was to determine the prevalence of Toxoplasma gondii in native donkeys in Mousl, Iraq. Fifty two sera (9 males and 43 females were examined by Latex agglutination test, Modified latex agglutination test with 2- mercaptoethanol test and Indirect ELISA test (Indirect IgG ELISA. The prevalence of Toxoplasma gondii in native donkeys was 46.15 %. Acute cases 8.33% and chronic cases 91.67 % when differentiated by Modified latex agglutination test with 2- mercaptoethanol test. The percentages of female and male infections were 51.16% (22/43 and 22.22% (2/9, respectively by using latex agglutination test, and the titeration of antibodies ranged between 1:20 - 1:1280 and for Indirect IgG ELISA it was 22.72% positive cases.

UNC93B1 mediates host resistance to infection with Toxoplasma gondii.

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Mariane B Melo

2010-08-01

Full Text Available UNC93B1 associates with Toll-Like Receptor (TLR 3, TLR7 and TLR9, mediating their translocation from the endoplasmic reticulum to the endolysosome, hence allowing proper activation by nucleic acid ligands. We found that the triple deficient '3d' mice, which lack functional UNC93B1, are hyper-susceptible to infection with Toxoplasma gondii. We established that while mounting a normal systemic pro-inflammatory response, i.e. producing abundant MCP-1, IL-6, TNFα and IFNγ, the 3d mice were unable to control parasite replication. Nevertheless, infection of reciprocal bone marrow chimeras between wild-type and 3d mice with T. gondii demonstrated a primary role of hemopoietic cell lineages in the enhanced susceptibility of UNC93B1 mutant mice. The protective role mediated by UNC93B1 to T. gondii infection was associated with impaired IL-12 responses and delayed IFNγ by spleen cells. Notably, in macrophages infected with T. gondii, UNC93B1 accumulates on the parasitophorous vacuole. Furthermore, upon in vitro infection the rate of tachyzoite replication was enhanced in non-activated macrophages carrying mutant UNC93B1 as compared to wild type gene. Strikingly, the role of UNC93B1 on intracellular parasite growth appears to be independent of TLR function. Altogether, our results reveal a critical role for UNC93B1 on induction of IL-12/IFNγ production as well as autonomous control of Toxoplasma replication by macrophages.

Diagnosis of pulmonary infection with Toxoplasma gondii in immunocompromised HIV-positive patients by real-time PCR

DEFF Research Database (Denmark)

Petersen, E; Edvinsson, B; Lundgren, B

2006-01-01

The aim of the study presented here was to evaluate the use of PCR for improving the diagnosis of Toxoplasma gondii infection in immunocompromised hosts. Three hundred thirty-two bronchoalveolar lavage (BAL) fluid samples were analyzed by real-time PCR targeting a 529 bp element of T. gondii. In ...

Metabolic Cooperation of Glucose and Glutamine Is Essential for the Lytic Cycle of Obligate Intracellular Parasite Toxoplasma gondii.

Science.gov (United States)

Nitzsche, Richard; Zagoriy, Vyacheslav; Lucius, Richard; Gupta, Nishith

2016-01-01

Toxoplasma gondii is a widespread protozoan parasite infecting nearly all warm-blooded organisms. Asexual reproduction of the parasite within its host cells is achieved by consecutive lytic cycles, which necessitates biogenesis of significant energy and biomass. Here we show that glucose and glutamine are the two major physiologically important nutrients used for the synthesis of macromolecules (ATP, nucleic acid, proteins, and lipids) in T. gondii, and either of them is sufficient to ensure the parasite survival. The parasite can counteract genetic ablation of its glucose transporter by increasing the flux of glutamine-derived carbon through the tricarboxylic acid cycle and by concurrently activating gluconeogenesis, which guarantee a continued biogenesis of ATP and biomass for host-cell invasion and parasite replication, respectively. In accord, a pharmacological inhibition of glutaminolysis or oxidative phosphorylation arrests the lytic cycle of the glycolysis-deficient mutant, which is primarily a consequence of impaired invasion due to depletion of ATP. Unexpectedly, however, intracellular parasites continue to proliferate, albeit slower, notwithstanding a simultaneous deprivation of glucose and glutamine. A growth defect in the glycolysis-impaired mutant is caused by a compromised synthesis of lipids, which cannot be counterbalanced by glutamine but can be restored by acetate. Consistently, supplementation of parasite cultures with exogenous acetate can amend the lytic cycle of the glucose transport mutant. Such plasticity in the parasite's carbon flux enables a growth-and-survival trade-off in assorted nutrient milieus, which may underlie the promiscuous survival of T. gondii tachyzoites in diverse host cells. Our results also indicate a convergence of parasite metabolism with cancer cells. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Seroprevalence of Toxoplasma gondii infection among pregnant women in Cameroon

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Anna L. Njunda

2011-09-01

Full Text Available Toxoplasmosis is caused by an intracellular protozoan, Toxoplasma gondii, which has a wide geographical distribution. The congenital form results in a gestational form that can present a temporary parasiteamia that will infect the fetus. For this reason early diagnosis in pregnancy is highly desirable, allowing prompt intervention in cases of infection. The aim of this study was to determine the seroprevalence of Toxoplasma gondii antibodies among pregnant women attending the Douala General Hospital. The study was carried out between March and July 2009, whereby 110 pregnant women were tested for IgG and IgM antibodies and information about eating habits and hygienic conditions was collected using a questionnaire. These women’s ages ranged from 20-44 years old with an average of 29.9 years; the overall IgG and IgM seroprevalence was 70% and 2.73 % respectively. Seroprevalence was significantly high amongst women who ate raw vegetables (76.39%, P<0.05 and there was a significant trend towards a higher seroprevalence in women who did not have a good source of water (75.58%, P<0.05. This research showed that consumption raw vegetables and poor quality drinking water are two risk factors associated with Toxoplasma gondii infection amongst pregnant women attending the Douala General Hospital in Cameroon.

Long-term humoral antibody responses by various serologic tests in pigs orally inoculated with oocysts of four strains of Toxoplasma gondii

DEFF Research Database (Denmark)

Dubey, J.P.; Andrews, C.D.; Thulliez, P.

1997-01-01

Antibody titers to Toxoplasma gondii were determined in 16 pigs orally inoculated with 1000 or 10000 oocysts of one of the four strains (GT-1, ME-49, TS-2, TC-2) of T. gondii. Pigs were euthanized on postinoculation days 103-875 and their tissues were bioassayed for T. gondii. Antibody titers wer...

Sero-prevalence of Toxoplasma gondii in Danish pigs

DEFF Research Database (Denmark)

Kofoed, Kristina Grønbech; Vorslund-Kiær, Mia; Nielsen, Henrik Vedel

2017-01-01

In 2015, the World Health Organisation rated toxoplasmosis as one of the most important food borne zoonotic diseases in the world. In addition, recent studies have associated Toxoplasma gondii sero-positivity with severe mental illnesses such as schizophrenia. Intake of raw or insufficiently cooked...

Carne de consumo e risco de transmissão de Toxoplasma gondii

OpenAIRE

Rosado, Joana de Carvalho Serra

2009-01-01

O consumo de carne de suíno (Sus domesticus) é considerado o maior factor de risco para a transmissão da toxoplasmose aos humanos. No entanto, apesar dos suínos serem muito usados na gastronomia portuguesa, tanto pelo consumo de carne, como pelo de enchidos, pouco se sabe sobre a prevalência e caracterização genética de Toxoplasma gondii nestes produtos alimentares, no país. Neste trabalho, pretendeu-se determinar a seroprevalência de anticorpos IgG anti-T. gondii em suínos de ...

Molecular characterization of resistance to Rifampicin in an emerging hospital-associated Methicillin-resistant Staphylococcus aureus clone ST228, Spain

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Liñares Josefina

2010-03-01

Full Text Available Abstract Background Methicillin-resistant S. aureus (MRSA has been endemic in Hospital Universitari de Bellvitge, Barcelona, since 1990. During the 1990-95 period the Iberian clone (ST-247; SCCmec-I was dominant. Isolates of clonal complex 5 (ST-125; SCCmec-IV gradually replaced the Iberian clone from 1996 to 2003. A new multiresistant MRSA phenotype showing rifampicin resistance emerged in 2004 and rapidly increased from 25% in 2004 to 45% in 2006. The aims of this study were i the molecular characterisation of rifampicin resistant MRSA isolates, ii the study of the rifampicin resistance expression by disk diffusion, microdilution and E-test, and iii the analysis of the rpoB gene mutations involved in rifampicin resistance. Results A sample of representative 108 rifampicin-resistant MRSA isolates belonged to a single PFGE genotype, ST-228, SCCmec type I and spa type t041. Of 108 isolates, 104 (96% had a low-level rifampicin resistance (MICs, 2 to 4 mg/L and 4 a high-level rifampicin resistance (MICs, 128 - ≥ 256 mg/L. Disk diffusion and E-test methods failed to identify a low-level rifampicin resistance in 20 and 12 isolates, respectively. A low-level rifampicin resistance was associated with amino acid substitution 481His/Asn in the beta-subunit of RNA polymerase. Isolates with a high-level rifampicin resistance carried additional mutations in the rpoB gene. Conclusions The emergence of MRSA clone ST228-SCCmecI, related to the Southern Germany clone, involved a therapeutical challenge for treating serious MRSA infections. Decreased susceptibility to rifampicin in MRSA strains of ST228-SCCmecI was associated with one or two specific mutations in the rpoB gene. One fifth of isolates with low-level rifampicin-resistance were missed by the diffusion methods.

Seroepidemiology of Toxoplasma gondii infection in drivers involved in road traffic accidents in the metropolitan area of Guadalajara, Jalisco, Mexico.

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Galván-Ramírez, Ma de la Luz; Sánchez-Orozco, Laura Verónica; Rodríguez, Laura Rocío; Rodríguez, Saúl; Roig-Melo, Enrique; Troyo Sanromán, Rogelio; Chiquete, Erwin; Armendáriz-Borunda, Juan

2013-10-11

The prevalence of toxoplasmosis in the general population of Guadalajara, Mexico, is around 32%. Toxoplasmosis can cause ocular lesions and slowing of reaction reflexes. Latent toxoplasmosis has been related with traffic accidents. We aimed to assess the prevalence of anti-Toxoplasma gondii antibodies and visual impairments related with traffic accidents in drivers from the metropolitan Guadalajara. We prospectively evaluated the prevalence of IgG and IgM anti-T. gondii antibodies in 159 individuals involved in traffic accidents, and in 164 control drivers never involved in accidents. Cases of toxoplasmosis reactivation or acute infection were detected by PCR in a subset of 71 drivers studied for the presence of T. gondii DNA in blood samples. Ophthalmologic examinations were performed in drivers with IgG anti-T. gondii antibodies in search of ocular toxoplasmosis. Fifty-four (34%) traffic accident drivers and 59 (36%) controls were positive to IgG anti-T. gondii antibodies (p = 0.70). Among the 113 seropositive participants, mean anti-T. gondii IgG antibodies titers were higher in traffic accident drivers than in controls (237.9 ± 308.5 IU/ml vs. 122.9 ± 112.7 IU/ml, respectively; p = 0.01 by Student's t test, p = 0.037 by Mann-Whitney U test). In multivariate analyses, anti-T. gondii IgG antibody titers were consistently associated with an increased risk of traffic accidents, whereas age showed an inverse association. The presence of IgM-anti-T. gondii antibodies was found in three (1.9%) subjects among traffic accident drives, and in two (1.2%) controls. Three (4.2%) samples were positive for the presence of T. gondii DNA, all among seropositive individuals. No signs of ocular toxoplasmosis were found in the entire cohort. Moreover, no other ocular conditions were found to be associated with the risk of traffic accidents in a multivariate analysis. Anti-T. gondii antibody titers are associated with the risk of traffic accidents. We could not determine any

Molecular cloning and functional analysis of the gene encoding ...

African Journals Online (AJOL)

Here we report for the first time the cloning of a full-length cDNA encoding GGPPS (Jc-GGPPS) from Jatropha curcas L. The full-length cDNA was 1414 base pair (bp), with an 1110-bp open reading frame (ORF) encoding a 370- amino-acids polypeptide. Bioinformatic analysis revealed that Jc-GGPPS is a member of the ...

New Toxoplasma gondii genotypes isolated from free-range chickens from the Fernando de Noronha, Brazil: unexpected findings

Science.gov (United States)

Worldwide comparison of Toxoplasma gondii isolates from free range chickens has indicated that T. gondii isolates from Brazil are phenotypically and genetically different than isolates from other countries; most strains from Brazil are pathogenic to mice, there is great genetic variability, most iso...

Climbing Mount Efficiency--small steps, not giant leaps towards higher cloning success in farm animals.

Science.gov (United States)

Oback, Björn

2008-07-01

Despite more than a decade of research efforts, farm animal cloning by somatic cell nuclear transfer (SCNT) is still frustratingly inefficient. Inefficiency manifests itself at different levels, which are currently not well integrated. At the molecular level, it leads to widespread genetic, epigenetic and transcriptional aberrations in cloned embryos. At the organismal level, these genome-wide abnormalities compromise development of cloned foetuses and offspring. Specific molecular defects need to be causally linked to specific cloned phenotypes, in order to design specific treatments to correct them. Cloning efficiency depends on the ability of the nuclear donor cell to be fully reprogrammed into an embryonic state and the ability of the enucleated recipient cell to carry out the reprogramming reactions. It has been postulated that reprogrammability of the somatic donor cell epigenome is influenced by its differentiation status. However, direct comparisons between cells of divergent differentiation status within several somatic lineages have found no conclusive evidence for this. Choosing somatic stem cells as donors has not improved cloning efficiency, indicating that donor cell type may be less critical for cloning success. Different recipient cells, on the other hand, vary in their reprogramming ability. In bovine, using zygotes instead of oocytes has increased cloning success. Other improvements in livestock cloning efficiency include better coordinating donor cell type with cell cycle stage and aggregating cloned embryos. In the future, it will be important to demonstrate if these small increases at every step are cumulative, adding up to an integrated cloning protocol with greatly improved efficiency.

Immunity Status of Blood Donors Regarding Toxoplasma gondii Infection in a Low-Income District of Abidjan, Côte d’Ivoire, West Africa

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Liliane Siransy

2016-01-01

Full Text Available Background. Toxoplasmosis is a widespread cosmopolitan anthropozoonosis, which affects more than a third of the world population. Except the modes of transmission well known, Toxoplasma gondii can be transmitted during transplantation or blood transfusion. The aim of this study is to determine the prevalence of IgG and IgM Toxoplasma gondii and to estimate the potential risk by blood products. Methods. This is a cross-sectional study on the research for Toxoplasma gondii antibodies (IgG and IgM blood donors performed by ELISA. Results. An overall seroprevalence of Toxoplasma gondii among blood donors recruited was 67.92% (n=72. Among these, 68 have Toxoplasma gondii IgG (64.15%, 12 Toxoplasma gondii IgM (11.32%, and 4 (3.77% both. The risk varies between 8 for 100000 and 172 for 100000 donations. Conclusion. The need to strengthen security measures for people multitransfused, immunocompromised, and pregnant women to reduce the transmission of toxoplasmosis is important.

Goats reinfected with Toxoplasma gondii: loss of viable prolificacy and gross revenue

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H. M. Silva

2015-10-01

Full Text Available ABSTRACTWe determined the reproductive parameters and clinical disorders in pregnant goats infected and reinfected with Toxoplasma gondii, and posteriorly the loss of gross revenue due to congenital toxoplasmosis was estimated. Of the 25 non-pregnant females negative for T. gondii, 20 were orally inoculated (ME 49 strain and of these, 15 pregnant females chronically infected were orally reinoculated (VEG strain with T. gondii oocysts. Five groups were formed (n=5: GI, GII and GIII (reinoculations at 40, 80 and 120 days of gestation, respectively, GIV (inoculation and GV (no inoculation. Clinical and serological exams were performed on days 0 (prior to inoculation, 3, 6 9, 15 and 21 and every 7 days post-inoculation. Exams were also performed on day 3 and every 7 days post-reinoculation. Reproductive management was performed on all females and initiated when the females infected displayed IgG titers IFAT<1,024. From the average prolificacy indexes of each experimental group were estimated: total production of kilograms of live weight (total kg LW of goats for slaughter, gross revenue and loss of gross revenue in U.S. dollars (US$, designed for a herd of 1,000 matrices. The unviable prolificacy indexes were 0.8 (GI, 1.2 (GII and 0.2 (GIII. Clinical disorders affected 57.1% (GI, 75.0% (GII and 16.7% (GIII of the offspring of goats reinfected with T. gondii. Congenital toxoplasmosis in goats reinfected resulted in the loss of 26.5% of gross revenues, being GI (US$ 10,577.60 or 57.1% and GII (US$ 12,693.12 or 60% holders of the highest values and percentages of economic losses. It was found that congenital toxoplasmosis reinfection cause clinical disorders in goats chronically infected with T. gondii and their offspring with birth of unviable animals and loss of gross revenue, at different stages of pregnancy (40, 80 and 120 days of gestation, being in the initial and intermediate stages of pregnancy the largest estimates of these losses.

Clone DB: an integrated NCBI resource for clone-associated data

Science.gov (United States)

Schneider, Valerie A.; Chen, Hsiu-Chuan; Clausen, Cliff; Meric, Peter A.; Zhou, Zhigang; Bouk, Nathan; Husain, Nora; Maglott, Donna R.; Church, Deanna M.

2013-01-01

The National Center for Biotechnology Information (NCBI) Clone DB (http://www.ncbi.nlm.nih.gov/clone/) is an integrated resource providing information about and facilitating access to clones, which serve as valuable research reagents in many fields, including genome sequencing and variation analysis. Clone DB represents an expansion and replacement of the former NCBI Clone Registry and has records for genomic and cell-based libraries and clones representing more than 100 different eukaryotic taxa. Records provide details of library construction, associated sequences, map positions and information about resource distribution. Clone DB is indexed in the NCBI Entrez system and can be queried by fields that include organism, clone name, gene name and sequence identifier. Whenever possible, genomic clones are mapped to reference assemblies and their map positions provided in clone records. Clones mapping to specific genomic regions can also be searched for using the NCBI Clone Finder tool, which accepts queries based on sequence coordinates or features such as gene or transcript names. Clone DB makes reports of library, clone and placement data on its FTP site available for download. With Clone DB, users now have available to them a centralized resource that provides them with the tools they will need to make use of these important research reagents. PMID:23193260

Aspectos ultraestruturais do processo de divisão do Toxoplasma gondii

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Wanderley de Souza

1974-02-01

Full Text Available Neste trabalho é feita uma revisão sobre alguns aspectos biológicos do Toxoplasma gondii, principalmente sobre a ultraestrutura da forma interfásica e as modificações ultraestruturais que ocorrem no parasito durante o seu processo de divisão. Considera-se inicialmente o processo de divisão binária admitindo-se, porém, a possibilidade de que as imagens interpretadas como senão de divisão binária representem estágios da divisão por endodiogenia. Quanto à endodiogenia descrevem-se as alterações que ocorrem na "parasito mãe" durante o processo de formação dos dois "parasitos filhos". Este processo é semelhante no Toxoplasma gondii, Besnoitia jellisoni, Sarcocystis tenella e Frenkelia. Discute-se a possibilidade da formação de mais de dois "parasitos filhos" por um processo de endopoligenia, bem como o processo de esquizogonia. Os resultados mais recentes mostram que não existe esquizogonia nas formas vsgetativas do Toxoplasma gondii, senão que as imagens interpretadas como tal, ao microscópio ótico, são o resultado de endodiogenias sucessivas em que os endozoitas formados permanecem ligados entre si pela região posterior. A esquizogonia é, no entanto, encontrada nas formas que se desenvolvem no interior de células epiteliais do intestino do gato, que é o hospedeiro definitivo do Toxoplasma gondii. Discute-se o conceito de esquizogonia, comparando-o em três protozoários: Eimeria bovis, E. callospermophili e Plasmodium juxtanucleare, que apresentam diferenças entre si quanto ao processo de iniciação da individualização dos "parasitos filhos". Refere-se à recente hipótese que considera a endodiogenia como o processo fundamental de divisão dos esporozoárlos, ocorrendo na fase final da esquizogonia. Finalmente é acentuado o papel que a microscopia eletrônica aliada às modernas técnicas de citoquímica e imunocitoquimica poderá desempenhar no sentido de um melhor conhecimento da biologia do Toxoplasma

Reproductive implications of exposure to Toxoplasma gondii and Neospora caninum in western grey kangaroos (Macropus fuliginosus ocydromus).

Science.gov (United States)

Mayberry, Chris; Maloney, Shane K; Mitchell, Jeff; Mawson, Peter R; Bencini, Roberta

2014-04-01

Australian marsupials are thought to be particularly vulnerable to pathologic impacts of Toxoplasma gondii, and they may be similarly affected by Neospora caninum. Pathology due to either organism could be expressed as reduced female reproductive performance. We studied adult female western grey kangaroos (Macropus fuliginosus ocydromus) from suburban Perth, Western Australia, between May 2006 and October 2008. We used indirect fluorescent antibody tests to look for evidence of exposure to T. gondii and N. caninum in M. fuliginosus ocydromus and tested the association between their reproductive performance and a positive test result. Although 20% of plasma samples collected from 102 female kangaroos were positive for T. gondii and 18% were positive for N. caninum, we found no association between positive results and reproductive performance. Further study will be required to clarify if, and under what circumstances, T. gondii and N. caninum are pathogenic to macropod marsupials.

Research Article Molecular cloning and mRNA expression pattern of ...

Indian Academy of Sciences (India)

SAMSUNG

homologue from the brain of Misgurnus anguillicaudatus using homologous cloning and ... developmental processes, including sex determination, embryonic stem cell ..... anguillicaudatus, possibly aiding in unravelling reproductive biology ... significant difference (p < 0.05) between female and male in the same tissue.

Cloning and sequencing of Lol pI, the major allergenic protein of rye-grass pollen.

Science.gov (United States)

Griffith, I J; Smith, P M; Pollock, J; Theerakulpisut, P; Avjioglu, A; Davies, S; Hough, T; Singh, M B; Simpson, R J; Ward, L D

1991-02-25

We have isolated a full length cDNA clone encoding the major glycoprotein allergen Lol pI. The clone was selected using a combination of immunological screening of a cDNA expression library and PCR amplification of Lol pI-specific transcripts. Lol pI expressed in bacteria as a fusion protein shows recognition by specific IgE antibodies present in sera of grass pollen-allergic subjects. Northern analysis has shown that the Lol pI transcripts are expressed only in pollen of rye-grass. Molecular cloning of Lol pI provides a molecular genetic approach to study the structure-function relationship of allergens.

Seroprevalence of Toxoplasma gondii in mainland and sub-Antarctic New Zealand sea lion (Phocarctos hookeri) populations.

Science.gov (United States)

Michael, S A; Howe, L; Chilvers, B L; Morel, Pch; Roe, W D

2016-09-01

To investigate the seroprevalence of antibodies to Toxoplasma gondii in New Zealand sea lions (Phocarctos hookeri), as a potential contributor to reproductive failure. Archived sera were sourced from New Zealand sea lions from two recolonising mainland populations in the Otago Peninsula (n=15) and Stewart Island (n=12), as well as a declining population at Enderby Island (n=28) in the New Zealand sub-Antarctic. Sera were tested for antibodies to T. gondii using a commercially available ELISA (with samples considered positive if the sample to positive ratio was >30%), and latex agglutination test (LAT; with titres ≥1:32 considered positive). Western blot analysis was used to validate the results of a subset of 14 samples. Five samples from sea lions in mainland locations were confirmed positive for antibodies to T. gondii. Two adult females exhibited high LAT antibody titres (min 1:2048, max 1:4096) on both occasions when sampled 1 and 2 years apart, respectively. No animals from Enderby Island were seropositive. Toxoplasma gondii infection is unlikely to be a major contributor to poor reproductive success in New Zealand sea lions. However, continued surveillance is pertinent to assess subclinical and clinical impacts of the parasite on these threatened populations. The commercial tests evaluated here, with further species-specific threshold refinement could provide a fast, inexpensive and reliable indicator of T. gondii exposure in New Zealand sea lions.

Portugal and Angola: similarities and differences in Toxoplasma gondii seroprevalence and risk factors in pregnant women.

Science.gov (United States)

Lobo, M L; Patrocinio, G; Sevivas, T; DE Sousa, B; Matos, O

2017-01-01

In this study we determined the presence of IgM/IgG antibodies to Toxoplasma gondii in sera of 155 and 300 pregnant women from Lisbon (Portugal) and Luanda (Angola), respectively, and evaluated the potential risk factors associated with this infection. DNA detection was performed by PCR assays targeting T. gondii regions (RE/B1). Overall, 21·9% (10·9% IgG, 10·9% IgG/IgM) of the Lisbon women and 27·3% (23·7%, IgG, 2% IgM, 1·7% IgG/IgM) of the Luanda women had antibodies to T. gondii. Single variable and binary logistic regression analyses were conducted. Based on the latter, contacts with cats (family/friends), and having more than two births were identified as risk factors for Toxoplasma infection in Lisbon women. In Luanda, the risk factors for T. gondii infection suggested by the single variable analysis (outdoor contact with cats and consumption of pasteurized milk/dairy products) were not confirmed by binary logistic regression. This study shows original data from Angola, and updated data from Portugal in the study of infection by T. gondii in pregnant women, indicating that the prevalence of anti-Toxoplasma antibodies is high enough to alert the government health authorities and implement appropriate measures to control this infection.

Seroprevalence and risk factors of Toxoplasma gondii infection ...

African Journals Online (AJOL)

The infection is also associated with human immune deficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). In Rwanda, the burden and risk factors of T. gondii infection among pregnant women and among HIV infected pregnant women is largely unknown. This cross-sectional study aimed at determining the ...

Anti-Toxoplasma gondii secretory IgA in tears of patients with ocular toxoplasmosis: immunodiagnostic validation by ELISA

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Maria Isabel Lynch

2009-09-01

Full Text Available Toxoplasma gondii causes posterior uveitis and the specific diagnosis is based on clinical criteria. The presence of anti-T. gondii secretory IgA (sIgA antibodies in patients' tears has been reported and an association was found between ocular toxoplasmosis and the anti-T. gondii sIgA isotype in Brazilian patients. The purpose of this study was to provide an objective validation of the published ELISA test for determining the presence of anti-T. gondii sIgA in the tears of individuals with ocular toxoplasmosis. Tears from 156 patients with active posterior uveitis were analysed; 82 of them presented characteristics of ocular toxoplasmosis (standard lesion and 74 patients presented uveitis due to other aetiologies. Cases of active posterior uveitis were considered standard when a new inflammatory focus satellite to old retinochoroidal scars was observed. The determination of anti-T. gondii sIgA was made using an ELISA test with crude tachyzoite antigenic extracts. Tears were collected without previous stimulation. Detection of sIgA showed 65.9% sensitivity (95% CI = 54.5-74.4, 71.6% specificity (95% CI = 59.8-81.2, a positive predictive value of 72% (95% CI = 60.3-81.5 and a negative predictive value of 65.4% (95% CI = 54.0-75.4. sIgA reactivity was higher in the tears of patients with active posterior uveitis due to T. gondii (p < 0.05. The test is useful for differentiating active posterior uveitis due to toxoplasmosis from uveitis caused by other diseases.

Anti-Toxoplasma gondii secretory IgA in tears of patients with ocular toxoplasmosis: immunodiagnostic validation by ELISA.

Science.gov (United States)

Lynch, Maria Isabel; Malagueño, Elizabeth; Lynch, Luiz Felipe; Ferreira, Silvana; Stheling, Raphael; Oréfice, Fernando

2009-09-01

Toxoplasma gondii causes posterior uveitis and the specific diagnosis is based on clinical criteria. The presence of anti-T. gondii secretory IgA (sIgA) antibodies in patients' tears has been reported and an association was found between ocular toxoplasmosis and the anti-T. gondii sIgA isotype in Brazilian patients. The purpose of this study was to provide an objective validation of the published ELISA test for determining the presence of anti-T. gondii sIgA in the tears of individuals with ocular toxoplasmosis. Tears from 156 patients with active posterior uveitis were analysed; 82 of them presented characteristics of ocular toxoplasmosis (standard lesion) and 74 patients presented uveitis due to other aetiologies. Cases of active posterior uveitis were considered standard when a new inflammatory focus satellite to old retinochoroidal scars was observed. The determination of anti-T. gondii sIgA was made using an ELISA test with crude tachyzoite antigenic extracts. Tears were collected without previous stimulation. Detection of sIgA showed 65.9% sensitivity (95% CI = 54.5-74.4), 71.6% specificity (95% CI = 59.8-81.2), a positive predictive value of 72% (95% CI = 60.3-81.5) and a negative predictive value of 65.4% (95% CI = 54.0-75.4). sIgA reactivity was higher in the tears of patients with active posterior uveitis due to T. gondii (p < 0.05). The test is useful for differentiating active posterior uveitis due to toxoplasmosis from uveitis caused by other diseases.

Distribution and Molecular Characterization of Acinetobacter baumannii International Clone II Lineage in Japan.

Science.gov (United States)

Matsui, Mari; Suzuki, Masato; Suzuki, Masahiro; Yatsuyanagi, Jun; Watahiki, Masanori; Hiraki, Yoichi; Kawano, Fumio; Tsutsui, Atsuko; Shibayama, Keigo; Suzuki, Satowa

2018-02-01

Multidrug-resistant (MDR) Acinetobacter spp. have been globally disseminated in association with the successful clonal lineage Acinetobacter baumannii international clone II (IC II). Because the prevalence of MDR Acinetobacter spp. in Japan remains very low, we characterized all Acinetobacter spp. ( n = 866) from 76 hospitals between October 2012 and March 2013 to describe the entire molecular epidemiology of Acinetobacter spp. The most prevalent species was A. baumannii ( n = 645; 74.5%), with A. baumannii IC II ( n = 245) accounting for 28.3% of the total. Meropenem-resistant isolates accounted for 2.0% ( n = 17) and carried IS Aba1-bla OXA-23-like ( n = 10), bla IMP ( n = 4), or IS Aba1-bla OXA-51-like ( n = 3). Multilocus sequence typing of 110 representative A. baumannii isolates revealed the considerable prevalence of domestic sequence types (STs). A. baumannii IC II isolates were divided into the domestic sequence type 469 (ST469) ( n = 18) and the globally disseminated STs ST208 ( n = 14) and ST219 ( n = 4). ST469 isolates were susceptible to more antimicrobial agents, while ST208 and ST219 overproduced the intrinsic AmpC β-lactamase. A. baumannii IC II and some A. baumannii non-IC II STs (e.g., ST149 and ST246) were associated with fluoroquinolone resistance. This study revealed that carbapenem-susceptible A. baumannii IC II was moderately disseminated in Japan. The low prevalence of acquired carbapenemase genes and presence of domestic STs could contribute to the low prevalence of MDR A. baumannii A similar epidemiology might have appeared before the global dissemination of MDR epidemic lineages. In addition, fluoroquinolone resistance associated with A. baumannii IC II may provide insight into the significance of A. baumannii epidemic clones. Copyright © 2018 American Society for Microbiology.

Molecular cloning of a Bangladeshi strain of very virulent infectious bursal disease virus of chickens and its adaptation in tissue culture by site-directed mutagenesis

International Nuclear Information System (INIS)

Islam, M.R.; Raue, R.; Mueller, H.

2005-01-01

Full-length cDNA of both genome segments of a Bangladeshi strain of very virulent infectious bursal disease virus (BD 3/99) were cloned in plasmid vectors along with the T7 promoter tagged to the 5'-ends. Mutations were introduced in the cloned cDNA to bring about two amino acid exchanges (Q253H and A284T) in the capsid protein VP2. Transfection of primary chicken embryo fibroblast cells with RNA transcribed in vitro from the full-length cDNA resulted in the formation of mutant infectious virus particles that grow in tissue culture. The pathogenicity of this molecularly-cloned, tissue-culture- adapted virus (BD-3tc) was tested in commercial chickens. The parental wild-type strain, BD 3/99, was included for comparison. The subclinical course of the disease and delayed bursal atrophy in BD-3tc-inoculated birds suggested that these amino acid substitutions made BD-3tc partially attenuated. (author)

Screening and identification of novel B cell epitopes of Toxoplasma gondii SAG1

OpenAIRE

Wang, Yanhua; Wang, Guangxiang; Zhang, Delin; Yin, Hong; Wang, Meng

2013-01-01

Background The identification of protein epitopes is useful for diagnostic purposes and for the development of peptide vaccines. In this study, the epitopes of Toxoplasma gondii SAG1 were identified using synthetic peptide techniques with the aid of bioinformatics. Findings Eleven peptides derived from T. gondii SAG1 were assessed by ELISA using pig sera from different time points after infection. Four (PS4, PS6, PS10 and PS11), out of the eleven peptides tested were recognized by all sera. T...

Molecular cloning and functional characterization of borneol dehydrogenase from the glandular trichomes of Lavandula x intermedia.

Science.gov (United States)

Sarker, Lukman S; Galata, Mariana; Demissie, Zerihun A; Mahmoud, Soheil S

2012-12-15

Several varieties of Lavandula x intermedia (lavandins) are cultivated for their essential oils (EOs) for use in cosmetic, hygiene and personal care products. These EOs are mainly constituted of monoterpenes including camphor, which contributes an off odor reducing the olfactory appeal of the oil. We have recently constructed a cDNA library from the glandular trichomes (the sites of EO synthesis) of L. x intermedia plants. Here, we describe the cloning of a borneol dehydrogenase cDNA (LiBDH) from this library. The 780 bp open reading frame of the cDNA encoded a 259 amino acid short chain alcohol dehydrogenase with a predicted molecular mass of ca. 27.5 kDa. The recombinant LiBDH was expressed in Escherichia coli, purified by Ni-NTA agarose affinity chromatography, and functionally characterized in vitro. The bacterially produced enzyme specifically converted borneol to camphor as the only product with K(m) and k(cat) values of 53 μM and 4.0 × 10(-4) s(-1), respectively. The LiBDH transcripts were specifically expressed in glandular trichomes of mature flowers indicating that like other Lavandula monoterpene synthases the expression of this gene is regulated in a tissue-specific manner. The cloning of LiBDH has far reaching implications in improving the quality of Lavandula EOs through metabolic engineering. Copyright © 2012 Elsevier Inc. All rights reserved.

Serological Survey and Factors Associated with Toxoplasma gondii Infection in Domestic Goats in Myanmar

Science.gov (United States)

Bawm, Saw; Maung, Wint Yi; Win, Myat Yee; Thu, May June; Chel, Hla Myet; Khaing, Tin Aye; Wai, Soe Soe; Htun, Lat Lat; Myaing, Tin Tin; Tiwananthagorn, Saruda; Igarashi, Makoto; Katakura, Ken

2016-01-01

Goat farming is important for the livelihood of millions of rural people because it contributes to food security and creation of assets. However, infection of goats with Toxoplasma gondii could be a source of parasite transmission to humans. The information on T. gondii infection of goat was not reported yet in Myanmar. A total of 119 goat serum samples were collected from three cities in the central region of Myanmar for T. gondii antibody survey. With the occurrence value obtained in this first study, a second one, more complete, with larger number (162) of animals and properties, was carried out and the risk factors and prevalence were determined. In both studies the samples were analyzed by the LAT. Of these, 32 (11.4%) samples were showed to be positive. The infection was associated with the presence of cats at the farm (odds ratio [OR] = 4.66, 95% confidential interval [CI] = 1.03–21.06), farming with different animal species (sheep, cattle, and pigs) (OR = 4.33, 95% CI = 1.57–11.94), and farming without good management practices (OR = 0.23, 95% CI = 0.06–0.83). This is the first T. gondii prevalence study in goats in the country. PMID:26904362

Serological Survey and Factors Associated with Toxoplasma gondii Infection in Domestic Goats in Myanmar

Directory of Open Access Journals (Sweden)

Saw Bawm

2016-01-01

Full Text Available Goat farming is important for the livelihood of millions of rural people because it contributes to food security and creation of assets. However, infection of goats with Toxoplasma gondii could be a source of parasite transmission to humans. The information on T. gondii infection of goat was not reported yet in Myanmar. A total of 119 goat serum samples were collected from three cities in the central region of Myanmar for T. gondii antibody survey. With the occurrence value obtained in this first study, a second one, more complete, with larger number (162 of animals and properties, was carried out and the risk factors and prevalence were determined. In both studies the samples were analyzed by the LAT. Of these, 32 (11.4% samples were showed to be positive. The infection was associated with the presence of cats at the farm (odds ratio [OR] = 4.66, 95% confidential interval [CI] = 1.03–21.06, farming with different animal species (sheep, cattle, and pigs (OR = 4.33, 95% CI = 1.57–11.94, and farming without good management practices (OR = 0.23, 95% CI = 0.06–0.83. This is the first T. gondii prevalence study in goats in the country.

Toxoplasma gondii infection induces dendritic retraction in basolateral amygdala accompanied by reduced corticosterone secretion

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Rupshi Mitra

2013-03-01

Pathological anxiety is thought to reflect a maladaptive state characterized by exaggerated fear. Naturally occurring perturbations that reduce fear can be crucial in the search for new treatments. The protozoan parasite Toxoplasma gondii invades rat brain and removes the fear that rats have of cat odors, a change believed to be parasitic manipulation of host behavior aimed at increasing parasite transmission. It is likely that mechanisms employed by T. gondii can be used as a heuristic tool to understand possible means of fear reduction in clinical settings. Male Long-Evans rats were infected with T. gondii and compared with sham-infected animals 8 weeks after infection. The amount of circulating plasma corticosterone and dendritic arborization of basolateral amygdala principal neurons were quantified. Previous studies have shown that corticosterone, acting within the basolateral amygdala, enhances the fear response to environmental stimuli. Here we show that T. gondii infection causes a dendritic retraction in basolateral amygdala neurons. Such dendritic retraction is accompanied by lower amounts of circulating corticosterone, both at baseline and when induced by an aversive cat odor. The concerted effects of parasitism on two pivotal physiological nodes of the fear response provide an animal model relevant to interactions between stress hormones and amygdalar plasticity.

Detection of Toxoplasma gondii DNA in horse meat from supermarkets in France and performance evaluation of two serological tests

OpenAIRE

Aroussi Abdelkrim; Vignoles Philippe; Dalmay François; Wimel Laurence; Dardé Marie-Laure; Mercier Aurélien; Ajzenberg Daniel

2015-01-01

In France, some cases of severe toxoplasmosis have been linked to the consumption of horse meat that had been imported from the American continent where atypical strains of Toxoplasma gondii are more common than in Europe. Many seroprevalence studies are presented in the literature but risk assessment of T. gondii infection after horse meat consumption is not possible in the absence of validated serological tests and the unknown correlation between detection of antibodies against T. gondii an...

Toxoplasma gondii Infection in Mice Impairs Long-Term Fear Memory Consolidation through Dysfunction of the Cortex and Amygdala.

Science.gov (United States)

Ihara, Fumiaki; Nishimura, Maki; Muroi, Yoshikage; Mahmoud, Motamed Elsayed; Yokoyama, Naoaki; Nagamune, Kisaburo; Nishikawa, Yoshifumi

2016-10-01

Chronic infection with Toxoplasma gondii becomes established in tissues of the central nervous system, where parasites may directly or indirectly modulate neuronal function. Epidemiological studies have revealed that chronic infection in humans is a risk factor for developing mental diseases. However, the mechanisms underlying parasite-induced neuronal dysfunction in the brain remain unclear. Here, we examined memory associated with conditioned fear in mice and found that T. gondii infection impairs consolidation of conditioned fear memory. To examine the brain pathology induced by T. gondii infection, we analyzed the parasite load and histopathological changes. T. gondii infects all brain areas, yet the cortex exhibits more severe tissue damage than other regions. We measured neurotransmitter levels in the cortex and amygdala because these regions are involved in fear memory expression. The levels of dopamine metabolites but not those of dopamine were increased in the cortex of infected mice compared with those in the cortex of uninfected mice. In contrast, serotonin levels were decreased in the amygdala and norepinephrine levels were decreased in the cortex and amygdala of infected mice. The levels of cortical dopamine metabolites were associated with the time spent freezing in the fear-conditioning test. These results suggest that T. gondii infection affects fear memory through dysfunction of the cortex and amygdala. Our findings provide insight into the mechanisms underlying the neurological changes seen during T. gondii infection. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Sequence Variation in Rhoptry Neck Protein 10 Gene among Toxoplasma gondii Isolates from Different Hosts and Geographical Locations

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Yu ZHAO

2017-09-01

Full Text Available Background: Toxoplasma gondii, as a eukaryotic parasite of the phylum Apicomplexa, can infect almost all the warm-blooded animals and humans, causing toxoplasmosis. Rhoptry neck proteins (RONs play a key role in the invasion process of T. gondii and are potential vaccine candidate molecules against toxoplasmosis.Methods: The present study examined sequence variation in the rhoptry neck protein 10 (TgRON10 gene among 10 T. gondii isolates from different hosts and geographical locations from Lanzhou province during 2014, and compared with the corresponding sequences of strains ME49 and VEG obtained from the ToxoDB database, using polymerase chain reaction (PCR amplification, sequence analysis, and phylogenetic reconstruction by Bayesian inference (BI and maximum parsimony (MP. Results: Analysis of all the 12 TgRON10 genomic and cDNA sequences revealed 7 exons and 6 introns in the TgRON10 gDNA. The complete genomic sequence of the TgRON10 gene ranged from 4759 bp to 4763 bp, and sequence variation was 0-0.6% among the 12 T. gondii isolates, indicating a low sequence variation in TgRON10 gene. Phylogenetic analysis of TgRON10 sequences showed that the cluster of the 12 T. gondii isolates was not completely consistent with their respective genotypes.Conclusion: TgRON10 gene is not a suitable genetic marker for the differentiation of T. gondii isolates from different hosts and geographical locations, but may represent a potential vaccine candidate against toxoplasmosis, worth further studies.

Sequence Variation in Rhoptry Neck Protein 10 Gene among Toxoplasma gondii Isolates from Different Hosts and Geographical Locations.

Science.gov (United States)

Zhao, Yu; Zhou, Donghui; Chen, Jia; Sun, Xiaolin

2017-01-01

Toxoplasma gondii, as a eukaryotic parasite of the phylum Apicomplexa, can infect almost all the warm-blooded animals and humans, causing toxoplasmosis. Rhoptry neck proteins (RONs) play a key role in the invasion process of T. gondii and are potential vaccine candidate molecules against toxoplasmosis. The present study examined sequence variation in the rhoptry neck protein 10 (TgRON10) gene among 10 T. gondii isolates from different hosts and geographical locations from Lanzhou province during 2014, and compared with the corresponding sequences of strains ME49 and VEG obtained from the ToxoDB database, using polymerase chain reaction (PCR) amplification, sequence analysis, and phylogenetic reconstruction by Bayesian inference (BI) and maximum parsimony (MP). Analysis of all the 12 TgRON10 genomic and cDNA sequences revealed 7 exons and 6 introns in the TgRON10 gDNA. The complete genomic sequence of the TgRON10 gene ranged from 4759 bp to 4763 bp, and sequence variation was 0-0.6% among the 12 T. gondii isolates, indicating a low sequence variation in TgRON10 gene. Phylogenetic analysis of TgRON10 sequences showed that the cluster of the 12 T. gondii isolates was not completely consistent with their respective genotypes. TgRON10 gene is not a suitable genetic marker for the differentiation of T. gondii isolates from different hosts and geographical locations, but may represent a potential vaccine candidate against toxoplasmosis, worth further studies.

Screening and identification of novel B cell epitopes of Toxoplasma gondii SAG1.

Science.gov (United States)

Wang, Yanhua; Wang, Guangxiang; Zhang, Delin; Yin, Hong; Wang, Meng

2013-04-30

The identification of protein epitopes is useful for diagnostic purposes and for the development of peptide vaccines. In this study, the epitopes of Toxoplasma gondii SAG1 were identified using synthetic peptide techniques with the aid of bioinformatics. Eleven peptides derived from T. gondii SAG1 were assessed by ELISA using pig sera from different time points after infection. Four (PS4, PS6, PS10 and PS11), out of the eleven peptides tested were recognized by all sera. Then, shorter peptides that were derived from PS4, PS6, PS10 and PS11 were predicted using bioinformatics and tested by experimentation. Four out of nine shorter peptides were identified successfully (amino acids 106-120, 166-180, 289-300 and 313-332). We have precisely located the epitopes of T. gondii SAG1 using pig sera collected at different time points after infection. The identified epitopes may be useful for the further study of epitope-based vaccines and diagnostic reagents.

Molecular cloning and functional analysis of the follicle-stimulating ...

African Journals Online (AJOL)

A 762 bp fragment of the 5'-flanking region of the FSHR gene from the Jintang black goat was cloned. The putative initial transcript site was the A at 681 bp and there were 7 putative cis-acting elements and 3 AT-rich regions. The sequence of the FSHR promoter from the Jintang black goat is 99.34% homology to Capra ...

Seroprevalence of Toxoplasma gondii and Neospora caninum in feral cats (Felis silvestris catus) in Majorca, Balearic Islands, Spain.

Science.gov (United States)

Millán, Javier; Cabezón, Oscar; Pabón, Marcela; Dubey, J P; Almería, Sonia

2009-11-12

Felids are important in the epidemiology of Toxoplasma gondii infection because they are the only hosts that can excrete environmentally resistant oocysts. Antibodies to T. gondii and Neospora caninum were determined in serum samples from 59 feral cats (Felis silvestris catus) captured in baited traps during authorized predator control campaigns in Majorca, Balearic Islands, Spain. Antibodies to T. gondii assayed by the modified agglutination test (MAT) were found in 50 (84.7%) of 59 cats with MAT titers of 1:25 in one cat, 1:200 in four cats, 1:500 in seven cats, 1:1000 in nine cats, and >or=1:2000 in 29 cats. Seroprevalence (MAT, 1:25 or more) was significantly higher in adults (94.6%) than in juveniles (cats in Majorca is one of the highest reported worldwide in this species and the highest observed in Europe to date. The results suggest that feral cats in Majorca have a high rate of T. gondii infection with important implications for public health on the island since the seropositive cats are likely to have already shed T. gondii oocysts in the environment.

Molecular cloning, characterization and developmental expression of porcine β-synuclein

DEFF Research Database (Denmark)

Larsen, Knud; Frandsen, Pernille Munk; Madsen, Lone Bruhn

2010-01-01

The synuclein family includes three known proteins: alpha-synuclein, beta-synuclein and gamma-synuclein. beta-Synuclein inhibits the aggregation of alpha-synuclein, a protein involved in Parkinson's disease. We have cloned and characterized the cDNA sequence for porcine beta-synuclein (SNCB) from...

The effects of quercetin towards reactive oxygen species levels and glutathione in Toxoplasma gondii profilin-exposed adipocytes in vitro

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Yulia D.S.

2018-04-01

Full Text Available Toxoplasma gondii (T. gondii has been found to potentially cause adipocyte dysfunction by activating the inflammatory pathways through its profilin. In response to inflammation, adipocytes produce Reactive Oxygen Species (ROS. To scavenge ROS, endogenous or exogenous antioxidants are required. Glutathione (GSH is one of enzimatic antioxidant that abundant in all of body cells. Quercetin, an exogenous antioxidant, can be widely found in natural products. This research aims to explore the effects of quercetin towards ROS and GSH stimulated from T. gondii profilin-exposed adipocytes. To achieve this, adipocytes were exposed to 20 µM T. gondii profilin and treated with four doses of quercetin; 31.25, 62.5, 125, and 250 µM. The results showed that quercetin significantly reduced the ROS levels (p <0,001 and significantly increased GSH (p <0,001 in T. gondii profilin-exposed adipocytes compared to untreated cells, with an effective dose of 62.5µM. This study implies that quercetin might be a promising candidate for development of antioxidant treatment interventions to prevent toxoplasmosis-mediated adipocytopathy.

Is Toxoplasma Gondii Infection Related to Brain and Behavior Impairments in Humans? Evidence from a Population-Representative Birth Cohort.

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Karen Sugden

Full Text Available Toxoplasma gondii (T. gondii is a protozoan parasite present in around a third of the human population. Infected individuals are commonly asymptomatic, though recent reports have suggested that infection might influence aspects of the host's behavior. In particular, Toxoplasma infection has been linked to schizophrenia, suicide attempt, differences in aspects of personality and poorer neurocognitive performance. However, these studies are often conducted in clinical samples or convenience samples.In a population-representative birth-cohort of individuals tested for presence of antibodies to T. gondii (N = 837 we investigated the association between infection and four facets of human behavior: neuropsychiatric disorder (schizophrenia and major depression, poor impulse control (suicidal behavior and criminality, personality, and neurocognitive performance. Suicide attempt was marginally more frequent among individuals with T. gondii seropositivity (p = .06. Seropositive individuals also performed worse on one out of 14 measures of neuropsychological function.On the whole, there was little evidence that T. gondii was related to increased risk of psychiatric disorder, poor impulse control, personality aberrations or neurocognitive impairment.

Molecular cloning of the Coch gene of guinea pig inner ear and its expression analysis in cultured fibrocytes of the spiral ligament.

Science.gov (United States)

Li, Lishu; Ikezono, Tetsuo; Sekine, Kuwon; Shindo, Susumu; Matsumura, Tomohiro; Pawankar, Ruby; Ichimiya, Issei; Yagi, Toshiaki

2010-08-01

We have cloned guinea pig Coch cDNA and the sequence information will be useful for future molecular study combined with physiological experiments. Proper Coch gene expression appears to be dependent on the unique extracellular micro-environment of the inner ear in vivo. These results provide insight into the Coch gene expression and its regulation. To characterize the guinea pig Coch gene, we performed molecular cloning and expression analysis in the inner ear and cultured fibrocytes of the spiral ligament. The Coch cDNA was isolated using RACE. Cochlin isofoms were studied by Western blot using three different types of mammalian inner ear. The cochlear fibrocytes were cultured and characterized by immunostaining. Coch gene expression in the fibrocytes was investigated and the influence of cytokine stimulation was evaluated. The full-length 1991 bp Coch cDNA that encodes a 553 amino acid protein was isolated. The sequence had significant homology with other mammals, and the sizes of the Cochlin isoforms were identical. In the cultured fibrocytes, Coch mRNA was expressed in a very small amount and the isoform production was different, compared with the results in vivo. Cytokine stimulation did not alter the level of mRNA expression or isoform formation.

Anti- toxoplasma gondii activity of constituents from Balsamocitrus ...

African Journals Online (AJOL)

Isolation, characterization and anti-Toxoplasma gondii activity of constituents from the CH2Cl2/MeOH (1/1) extract of the roots of the cameroonian plant Balsamocitrus camerunensis L. were investigated in this study. Four known coumarins derivatives were isolated, namely, marmin (1), imperatorin (2), xanthoxyletin (3), ...

Low titer of antibody against Toxoplasma gondii may be related to resistant to cancer

OpenAIRE

Maryam Sharafi Seyedeh; Salehi Nahid; Mortazavi Nahid; Danesh Pour Shima; Yousefi Morteza; Yousofi Darani Hossein

2015-01-01

Context: Toxoplasma gondii is a protozoan parasite with a world-wide distribution. However, the majority of infected cases remain symptomless. There are raising scientific evidences indicating that parasitic infections induce antitumor activity against certain types of cancers. The inhibitory effect of T. gondii on cancer growth has also been shown in cell culture and mouse model. Aims: Considering the anti-tumor effect of this parasite, in this study the relationship between low titer of ...

Concentration and retention of Toxoplasma gondii oocysts by marine snails demonstrate a novel mechanism for transmission of terrestrial zoonotic pathogens in coastal ecosystems

Science.gov (United States)

Krusor, Colin; Smith, Woutrina A.; Tinker, M. Tim; Silver, Mary; Conrad, Patricia A.; Shapiro, Karen

2015-01-01

The parasite Toxoplasma gondii is an environmentally persistent pathogen that can cause fatal disease in humans, terrestrial warm-blooded animals and aquatic mammals. Although an association between T. gondii exposure and prey specialization on marine snails was identified in threatened California sea otters, the ability of kelp-dwelling snails to transmit terrestrially derived pathogens has not been previously investigated. The objective of this study was to measure concentration and retention of T. gondii by marine snails in laboratory aquaria, and to test for natural T. gondii contamination in field-collected snails. Following exposure to T. gondii-containing seawater, oocysts were detected by microscopy in snail faeces and tissues for 10 and 3 days respectively. Nested polymerase chain reaction was also applied as a method for confirming putative T. gondii oocysts detected in snail faeces and tissues by microscopy. Toxoplasma gondiiwas not detected in field-collected snails. Results suggest that turban snails are competent transport hosts for T. gondii. By concentrating oocysts in faecal pellets, snails may facilitate entry of T. gondii into the nearshore marine food web. This novel mechanism also represents a general pathway by which marine transmission of terrestrially derived microorganisms can be mediated via pathogen concentration and retention by benthic invertebrates.

A new set of primers for the detection of Toxoplasma gondii in amniotic fluid using polymerase chain reaction.

Science.gov (United States)

Pelloux, H; Weiss, J; Simon, J; Muet, F; Fricker-Hidalgo, H; Goullier-Fleuret, A; Ambroise-Thomas, P

1996-04-15

A new PCR system including a pair of primers, a probe and an internal control were designed from the B1 gene of Toxoplasma gondii. The system described allowed the detection of less than 10 tachyzoites of the RH strain of T. gondii. Among 21 amniotic fluid samples, this system diagnosed the cases of congenital toxoplasmosis which were simultaneously diagnosed using mice inoculation, in vitro culture, and serology from both amniotic fluid and fetal blood. These results show that these new primers allow for a highly sensitive detection of T. gondii DNA.

Recombinational Cloning Using Gateway and In-Fusion Cloning Schemes

Science.gov (United States)

Throop, Andrea L.; LaBaer, Joshua

2015-01-01

The comprehensive study of protein structure and function, or proteomics, depends on the obtainability of full-length cDNAs in species-specific expression vectors and subsequent functional analysis of the expressed protein. Recombinational cloning is a universal cloning technique based on site-specific recombination that is independent of the insert DNA sequence of interest, which differentiates this method from the classical restriction enzyme-based cloning methods. Recombinational cloning enables rapid and efficient parallel transfer of DNA inserts into multiple expression systems. This unit summarizes strategies for generating expression-ready clones using the most popular recombinational cloning technologies, including the commercially available Gateway® (Life Technologies) and In-Fusion® (Clontech) cloning technologies. PMID:25827088

97 original article toxoplasma gondii infection in hiv/aids: prevalence

African Journals Online (AJOL)

Keywords: Toxoplasma gondii , IgG, Seroprevalence, HIV positive, CD4 cells. ... with immunosuppressive cancer and transplant ... due to active parasitaemia during pregnancy can .... especially in the night where they look for leftover.

Prevalence of Toxoplasma gondii in small mammals from the Ardennes region, France.

Science.gov (United States)

Afonso, Eve; Poulle, Marie-Lazarine; Lemoine, Mélissa; Villena, Isabelle; Aubert, Dominique; Gilot-Fromont, Emmanuelle

2007-11-01

Serum samples from 218 small mammals trapped in forest and grassland in the Ardennes region (North-eastern France) were tested for antibodies to Toxoplasma gondii. Using the modified agglutination test, positive results were found in 4/92 Apodemus sp., 3/64 Clethrionomys glareolus, 0/26 Microtus agrestis, 0/4 Micromys minutus, 3/5 Sorex sp., 2/9 Arvicola terrestris, and 7/18 Talpa europaea. Toxoplasma gondii was not isolated from the heart of seropositive individuals after bioassay in mice. Seroprevalence was significantly higher in large fossorial mammals living in grassland than in small forest mammals, probably related to ecological factors.

Toxoplasma gondii: prevalence and characterization of new genotypes in free-range chickens from south Brazil.

Science.gov (United States)

Vieira, Fernando Emmanuel Gonçalves; Sasse, João Pedro; Minutti, Ana Flávia; Miura, Ana Carolina; de Barros, Luiz Daniel; Cardim, Sergio Tosi; Martins, Thais Agostinho; de Seixas, Mércia; Yamamura, Milton Issashi; Su, Chunlei; Garcia, João Luis

2018-03-01

Toxoplasma gondii is an intracellular parasite that can infect all warm-blooded animals including humans. Recent studies showed that T. gondii strains from South America are genetically diverse. The present work aimed to determine T. gondii prevalence in free-ranging chicken in northwest Parana state in Brazil by two serological tests, to isolate the parasites from seropositive chickens and to genotype the isolates. Antibodies to T. gondii in 386 serum samples from 24 farms were investigated by immunofluorescence antibody assay (IFA) and modified agglutination test (MAT). Samples having titers ≥ 16 were considered positive for both tests. Among the 386 serum samples, 102 (26.4%) were positive for IFA, 64 (16.6%) were positive for MAT, 47 (12.2%) were positive in both tests, and 119 (30.8%) were positive in at least one of the two tests. Brain and pool of heart, lung, and liver from the 119 seropositive chickens were used for mouse bioassay to isolate the parasites. Thirty eight (31.9%) of these seropositive chickens were considered positives in mouse bioassay and 18 isolates were obtained. The isolates were characterized by 10 PCR-RFLP genetic markers including SAG1, SAG2 (5'-3'SAG2, alt.SAG2), SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico. Results of genotyping were compared with the genotypes in ToxoDB database. It revealed ten genotypes, including ToxoDB PCR-RFLP genotypes #6 (n = 2), #19 (n = 1), #21 (n = 2), #111 (n = 2), #152 (n = 1), and #175 (n = 1) and four new types not described before. Our results confirmed a high genetic diversity of this parasite in southern Brazil and also showed that the use of two serological tests in combination can improve the chance of T. gondii isolation. More studies should be taken to determine the zoonotic potential of chickens in the transmission of T. gondii.

Latitudinal variability in the seroprevalence of antibodies against Toxoplasma gondii in non-migrant and Arctic migratory geese

NARCIS (Netherlands)

Sandstrom, Cecilia A. M.; Buma, Anita G. J.; Hoye, Bethany J.; Prop, Jouke; van der Jeugd, Henk; Voslamber, Berend; Madsen, Jesper; Loonen, Maarten J. J. E.

2013-01-01

Toxoplasma gondii is an intracellular coccidian parasite found worldwide and is known to infect virtually all warm-blooded animals. It requires a cat (family Felidae) to complete its full life cycle. Despite the absence of wild felids on the Arctic archipelago of Svalbard, T. gondii has been found

[Molecular cloning, expression and characterization of lysine decarboxylase gene of endophytic fungus Shiraia sp. Slf14 from Huperzia serrata].

Science.gov (United States)

Peng, Silu; Yang, Huilin; Zhu, Du; Zhang, Zhibin; Yan, Riming; Wang, Ya

2016-04-14

Huperzine A (HupA) was approved as a drug for the treatment of Alzheimer's disease. The HupA biosynthetic pathway was started from lysine decarboxylase (LDC), which catalyzes lysine to cadaverine. In this study, we cloned and expressed an LDC gene from a HupA-producing endophytic fungus, and tested LDC activities. An endophytic fungus Shiraia sp. Slf14 from Huperzia serrata was used. LDC gene was obtained by RT-PCR, and cloned into pET-22b(+) and pET-32a(+) vectors to construct recombinant plasmids pET- 22b-LDC and pET-32a-LDC. These two recombinant plasmids were transformed into E. coli BL21, cultured for 8 h at 24 °C, 200 r/min with 1×10–3 mol/L IPTG into medium to express the LDC proteins, respectively. LDC proteins were purified by Ni2+ affinity chromatography. Catalytic activities were measured by Thin Layer Chromatography. At last, the physicochemical properties and structures of these two LDCs were obtained by bioinformatics software. LDC and Trx-LDC were expressed in E. coli BL21 successfully. SDS-PAGE analysis shows that the molecular weight of LDC and Trx-LDC were 24.4 kDa and 42.7 kDa respectively, which are consistent with bioinformatics analysis. In addition, TLC analysis reveals that both LDC and Trx-LDC had catalytic abilities. This work can provide fundamental data for enriching LDC molecular information and reveal the HupA biosynthetic pathway in endophytic fungi.

The Evolution of Host Mitochondrial Association and its Impact on Toxoplasma gondii Infection

Science.gov (United States)

English, Elizabeth D.

The association of intracellular pathogens with host mitochondria has been observed across taxa, from bacterial pathogens, such as Legionella pneumophila and Chlamydia trachomati, to the eukaryotic pathogen Toxoplasma gondii. However the functional impact of host mitochondrial association (HMA) remains difficult to assess in most of these species because in many cases the genes responsible for this phenomenon have not yet been identified. The recent discovery of the T. gondii gene responsible for HMA, Mitochondrial Association Factor 1 ( MAF1) has provided us with the tools to begin to understand the evolution and impact of HMA. Here we use multispecies sequence analysis to determine that the MAF1 locus is tandemly duplicated and diversified in both T. gondii and its nearest extant relative Hammondia hammondi, but not another close relative Neospora caninum. Using cross-species complementation we find that T. gondii and H. hammondi harbor copies of MAF1 able to mediate HMA, while N. caninum does not. We have begun mutational analysis using naturally occurring HMA+ and HMA- paralogs of MAF1 in order to determine the portions of MAF1 protein necessary for HMA. Additionally, we have identified the first in vivo phenotypes associated with HMA using multiple mouse models, for both acute and chronic infection. Taken together these data indicate that HMA likely evolved via neofunctionalization of a duplicated ancestral MAF1 gene, and that the neofunctionalized, HMA competent copy of MAF1 provides a selective advantage.

Toxoplasma gondii infection induces suppression in a mouse model of allergic airway inflammation.

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Ignacio M Fenoy

Full Text Available Allergic asthma is an inflammatory disorder characterized by infiltration of the airway wall with inflammatory cells driven mostly by activation of Th2-lymphocytes, eosinophils and mast cells. There is a link between increased allergy and a reduction of some infections in Western countries. Epidemiological data also show that respiratory allergy is less frequent in people exposed to orofecal and foodborne microbes such as Toxoplasma gondii. We previously showed that both acute and chronic parasite T. gondii infection substantially blocked development of airway inflammation in adult BALB/c mice. Based on the high levels of IFN-γ along with the reduction of Th2 phenotype, we hypothesized that the protective effect might be related to the strong Th1 immune response elicited against the parasite. However, other mechanisms could also be implicated. The possibility that regulatory T cells inhibit allergic diseases has received growing support from both animal and human studies. Here we investigated the cellular mechanisms involved in T. gondii induced protection against allergy. Our results show for the first time that thoracic lymph node cells from mice sensitized during chronic T. gondii infection have suppressor activity. Suppression was detected both in vitro, on allergen specific T cell proliferation and in vivo, on allergic lung inflammation after adoptive transference from infected/sensitized mice to previously sensitized animals. This ability was found to be contact-independent and correlated with high levels of TGF-β and CD4(+FoxP3(+ cells.

Chronic Toxoplasma gondii in Nurr1-Null Heterozygous Mice Exacerbates Elevated Open Field Activity

OpenAIRE

Eells, Jeffrey B.; Varela-Stokes, Andrea; Guo-Ross, Shirley X.; Kummari, Evangel; Smith, Holly M.; Cox, Erin; Lindsay, David S.

2015-01-01

Latent infection with Toxoplasma gondii is common in humans (approximately 30% of the global population) and is a significant risk factor for schizophrenia. Since prevalence of T. gondii infection is far greater than prevalence of schizophrenia (0.5-1%), genetic risk factors are likely also necessary to contribute to schizophrenia. To test this concept in an animal model, Nurr1-null heterozygous (+/-) mice and wild-type (+/+) mice were evaluate using an emergence test, activity in an open fie...

Occurrence of infection by Toxoplasma gondii in slaughtered swine in the northwestern region of Paraná, Brazil

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Hannah Lia Ettiene Peruch Lemos dos Santos

2015-07-01

Full Text Available This study aimed to determine the prevalence of Toxoplasma gondii in swine slaughtered in Iporã, Northwest Paraná state. Blood samples were obtained from 500 finishing swine. All animals were raised under intensive farming; the blood samples were analyzed using the immunofluorescence antibody test (IFAT. Animals with titers of ?64 were considered positive for T. gondii infection. Of the tested samples, 63 (12.6% were positive by IFAT; 58 (92.06% of these showed titers of 64 (4.7%, with titers 256 and two (3.1% titers of 1024. These pigs might be considered a source of T. gondii infection for humans

Mast cell activator compound 48/40 is not an effective adjuvant for UV-attenuated Toxoplasma gondii vaccine.

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Li, Xi; Chen, Shengjie; Huang, Shiguang; Lu, Fangli

2017-08-01

Toxoplasma gondii (T. gondii, Tg) is a globally distributed parasitic protozoan causing different forms of toxoplasmosis in humans. Mast cells (MCs) play a role during T. gondii infection. Several studies suggest that MC activator compound 48/80 (C48/80) may be an effective vaccine adjuvant resulting in a potent and protective antigen-specific immune response against bacteria or virus infections. The present study was performed to determine whether C48/80 had adjuvant activity for ultraviolet (UV)-attenuated T. gondii vaccine to induce protective immune responses against T. gondii in mouse model. Kunming mice were divided into the following groups: naive mice, naive mice administrated with C48/80 intraperitoneal (i.p.) injection, mice infected by i.p. injection of 10 4 T. gondii RH strain alone (Tg group), mice infected with 10 4 RH tachyzoites plus C48/80 administration (Tg + C48/80), mice immunized with UV-Tg alone, and mice immunized with UV-Tg plus C48/80 administration (UV-Tg + C48/80). All the vaccinated mice were challenged with 10 4 tachyzoites of T. gondii RH strain at the same time as the primary infection. The survival rates, liver histopathologies, liver parasite burdens, and mRNA expression levels of Th1 and Th2 cytokines in the livers and spleens detected by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) were compared among the aforementioned groups after primary infection or challenge infection. The results showed that, compared to the Tg group or Tg + C48/80 group, the UV-Tg + Tg group and UV-Tg + C48/80 + Tg group had significantly prolonged survival time, lower liver histopathological scores, decreased liver parasite burdens, and increased levels of Th1 and Th2 cytokines in the livers and spleens. There was no significant difference of survival time between the UV-Tg + Tg group and the UV-Tg + C48/80 + Tg group; however, the UV-Tg + C48/80 + Tg group showed higher parasite burden, more severe

A systematic review and meta-analysis of the seroprevalence of Toxoplasma gondii in cats in mainland China.

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Ding, Huan; Gao, Yu-Meng; Deng, Yao; Lamberton, Poppy H L; Lu, Da-Bing

2017-01-13

Toxoplasmosis is caused by Toxoplasma gondii which can infect all warm-blooded animals. As the most common feline definitive host, cats play a vital role in the transmission of T. gondii. However, national estimates of the seroprevalence of T. gondii in cats in mainland China are lacking, and therefore a systematic review and meta-analysis were performed to provide insight into national environmental transmission levels and potential transmission to humans. Studies published up until July 1, 2016, on T. gondii seroprevalence in cats within mainland China were searched for in CNKI, WanFang, CBM, PubMed, Embase and through the reference lists of resulting articles. The seroprevalence with its 95% confidence interval (CI) for each individual study was presented, and then point estimates and their 95% confidence intervals (CIs) of pooled seroprevalence were calculated. Subgroup analyses were performed according to potential risk factors. A total of 38 eligible studies, published between 1995 to 2016, covering fifteen provinces and municipalities, and involving 7,285 cats, were included. The seroprevalence in cats per study ranged from 3.9 to 79.4% with a median of 20.3%. As substantial heterogeneity existed among studies, a random-effects model was used to estimate the pooled seroprevalence. The value of the point estimate seroprevalence was 24.5% (95% CI: 20.1-29.0). Seroprevalence in stray cats was significantly higher than in pet cats (OR = 3.00, 95% CI: 1.60-5.64). The seroprevalence increased significantly with cat age (P = 0.018) with 17.4% (95% CI: 7.6-27.2) in the group of ≤ 1 year old, 19.5% (95% CI: 12.7-26.3) in the group of ≤ 3 year-old and 31.6% (95% CI: 22.9-40.3) in the group of > 3 year-old. The seroprevalence of T. gondii in cats in mainland China was moderate and was associated with cat ownership and age. Due to the increasing prevalence of pet cats in China and the intimate relationship between these cats and humans, this

Molecular cloning of chicken metallothionein. Deduction of the complete amino acid sequence and analysis of expression using cloned cDNA

Energy Technology Data Exchange (ETDEWEB)

Wei, D; Andrews, G K

1988-01-25

A cDNA library was constructed using RNA isolated from the livers of chickens which had been treated with zinc. This library was screened with a RNA probe complementary to mouse metallothionein-I (MT), and eight chicken MT cDNA clones were obtained. All of the cDNA clones contained nucleotide sequences homologous to regions of the longest (375 bp) cDNA clone. The latter contained an open reading frame of 189 bp, and the deduced amino acid sequence indicates a protein of 63 amino acids of which 20 are cysteine residues. Amino acid composition and partial amino acid sequence analyses of purified chicken MT protein agreed with the amino acid composition and sequence deduced from the cloned cDNA. Amino acid sequence comparison establish that chicken MT shares extensive homology with mammalian MTs. Southern blot analysis of chicken DNA indicates that the chicken MT gene is not a part of a large family of related sequences, but rather is likely to be a unique gene sequence. In the chicken liver, levels of chicken MT mRNA were rapidly induced by metals (Cd/sup 2 +/, Zn/sup 2 +/, Cu/sup 2 +/), glucocorticoids and lipopolysaccharide. MT mRNA was present in low levels in embryonic liver and increased to high levels during the first week after hatching before decreasing again to the basal levels found in adult liver. The results of this study establish that MT is highly conserved between birds and mammals and is regulated in the chicken by agents which also regulate expression of mammalian MT genes. However, in contrast to the mammals, the results suggest the existence of a single isoform of MT in the chicken.

Unified Approach to Universal Cloning and Phase-Covariant Cloning

OpenAIRE

Hu, Jia-Zhong; Yu, Zong-Wen; Wang, Xiang-Bin

2008-01-01

We analyze the problem of approximate quantum cloning when the quantum state is between two latitudes on the Bloch's sphere. We present an analytical formula for the optimized 1-to-2 cloning. The formula unifies the universal quantum cloning (UQCM) and the phase covariant quantum cloning.

Genotyping of polymorphic effectors of Toxoplasma gondii isolates from China

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Weisheng Cheng

2017-11-01

Full Text Available Abstract Background Toxoplasma gondii is an opportunistic protozoan apicomplexan and obligate intracellular parasite that infects a wide range of animals and humans. Rhoptry proteins 5 (ROP5, ROP16, ROP18 and dense granules 15 (GRA15 are the important effectors secreted by T. gondii which link to the strain virulence for mice and modulate the host’s response to the parasite. Little has been known about these molecules as well as GRA3 in type Chinese 1 strains that show polymorphism among strains of archetypical genotypes. This study examined the genetic diversity of these effectors and its correlated virulence in mice among T. gondii isolates from China. Results Twenty-one isolates from stray cats were detected, of which 15 belong to Chinese 1, and 6 to ToxoDB #205. Wh6 isolate, a Chinese 1 strain, has an avirulent phenotype. PCR-RFLP results of ROP5 and ROP18 presented few variations among the strains. Genotyping of GRA15 and ROP16 revealed that all the strains belong to type II allele except Xz7 which carries type I allele. ROP16 amino acid alignment at 503 locus demonstrated that 17 isolates are featured as type I or type III (ROP16I/III, and the other 4 as type II (ROP16II. The strains investigated may be divided into four groups based on GRA3 amino acid alignment, and all isolates of type Chinese 1 belong to the μ-1 allele except Wh6 which is identical to type II strain. Conclusions PCR-RFLP and sequence alignment analyses of ROP5, ROP16, ROP18, GRA3, and GRA15 in T. gondii revealed that strains with the same genotype may have variations in some of their key genes. GRA3 variation exhibited by Wh6 strain may be associated with the difference in phenotype and pathogenesis.

Polyparasitism Is Associated with Increased Disease Severity in Toxoplasma gondii-Infected Marine Sentinel Species

Science.gov (United States)

Gibson, Amanda K.; Raverty, Stephen; Lambourn, Dyanna M.; Huggins, Jessica; Magargal, Spencer L.; Grigg, Michael E.

2011-01-01

In 1995, one of the largest outbreaks of human toxoplasmosis occurred in the Pacific Northwest region of North America. Genetic typing identified a novel Toxoplasma gondii strain linked to the outbreak, in which a wide spectrum of human disease was observed. For this globally-distributed, water-borne zoonosis, strain type is one variable influencing disease, but the inability of strain type to consistently explain variations in disease severity suggests that parasite genotype alone does not determine the outcome of infection. We investigated polyparasitism (infection with multiple parasite species) as a modulator of disease severity by examining the association of concomitant infection of T. gondii and the related parasite Sarcocystis neurona with protozoal disease in wild marine mammals from the Pacific Northwest. These hosts ostensibly serve as sentinels for the detection of terrestrial parasites implicated in water-borne epidemics of humans and wildlife in this endemic region. Marine mammals (151 stranded and 10 healthy individuals) sampled over 6 years were assessed for protozoal infection using multi-locus PCR-DNA sequencing directly from host tissues. Genetic analyses uncovered a high prevalence and diversity of protozoa, with 147/161 (91%) of our sampled population infected. From 2004 to 2009, the relative frequency of S. neurona infections increased dramatically, surpassing that of T. gondii. The majority of T. gondii infections were by genotypes bearing Type I lineage alleles, though strain genotype was not associated with disease severity. Significantly, polyparasitism with S. neurona and T. gondii was common (42%) and was associated with higher mortality and more severe protozoal encephalitis. Our finding of widespread polyparasitism among marine mammals indicates pervasive contamination of waterways by zoonotic agents. Furthermore, the significant association of concomitant infection with mortality and protozoal encephalitis identifies polyparasitism as

Polyparasitism is associated with increased disease severity in Toxoplasma gondii-infected marine sentinel species.

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Amanda K Gibson

2011-05-01

Full Text Available In 1995, one of the largest outbreaks of human toxoplasmosis occurred in the Pacific Northwest region of North America. Genetic typing identified a novel Toxoplasma gondii strain linked to the outbreak, in which a wide spectrum of human disease was observed. For this globally-distributed, water-borne zoonosis, strain type is one variable influencing disease, but the inability of strain type to consistently explain variations in disease severity suggests that parasite genotype alone does not determine the outcome of infection. We investigated polyparasitism (infection with multiple parasite species as a modulator of disease severity by examining the association of concomitant infection of T. gondii and the related parasite Sarcocystis neurona with protozoal disease in wild marine mammals from the Pacific Northwest. These hosts ostensibly serve as sentinels for the detection of terrestrial parasites implicated in water-borne epidemics of humans and wildlife in this endemic region. Marine mammals (151 stranded and 10 healthy individuals sampled over 6 years were assessed for protozoal infection using multi-locus PCR-DNA sequencing directly from host tissues. Genetic analyses uncovered a high prevalence and diversity of protozoa, with 147/161 (91% of our sampled population infected. From 2004 to 2009, the relative frequency of S. neurona infections increased dramatically, surpassing that of T. gondii. The majority of T. gondii infections were by genotypes bearing Type I lineage alleles, though strain genotype was not associated with disease severity. Significantly, polyparasitism with S. neurona and T. gondii was common (42% and was associated with higher mortality and more severe protozoal encephalitis. Our finding of widespread polyparasitism among marine mammals indicates pervasive contamination of waterways by zoonotic agents. Furthermore, the significant association of concomitant infection with mortality and protozoal encephalitis identifies

Molecular Cloning and Characterization of Carbonic Anhydrase XII from Pufferfish (Takifugu rubripes

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Kanij Rukshana Sumi

2018-03-01

Full Text Available In this study, an 1888-bp carbonic anhydrase XII (CA XII sequence was cloned from the brain of the pufferfish, Takifugu rubripes. The cloned sequence contained a coding region of 1470-bp, which was predicted to translate into a protein of 490 amino acid residues. The predicted protein showed between 68–56% identity with the large yellow croaker (Larimichthys crocea, tilapia (Oreochromis niloticus, and Asian arowana (Scleropages formosus CA XII proteins. It also exhibited 36% and 53% identity with human CA II and CA XII, respectively. The cloned sequence contained a 22 amino acid NH2-terminal signal sequence and three Asn-Xaa-Ser/Thr sequons, among which one was potentially glycosylated. Four cysteine residues were also identified (Cys-21, Cys-201, Cys-355, and Cys-358, two of which (Cys-21 and Cys-201 could potentially form a disulfide bond. A 22-amino acid COOH-terminal cytoplasmic tail containing a potential site for phosphorylation by protein kinase A was also found. The cloned sequence might be a transmembrane protein, as predicted from in silico and phylogenetic analyses. The active site analysis of the predicted protein showed that its active site residues were highly conserved with tilapia CA XII protein. Homology modeling of the pufferfish CA XII was done using the crystal structure of the extracellular domain of human carbonic anhydrase XII at 1.55 Å resolution as a template. Semi-quantitative reverse transcription (RT-PCR, quantitative PCR (q-PCR, and in situ hybridization confirmed that pufferfish CA XII is highly expressed in the brain.

Soroprevalência de Toxoplasma gondii em rebanhos caprinos no Estado de São Paulo Seroprevalence of Toxoplasma gondii in dairy goats in the São Paulo State, Brazil

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Rodrigo Soares Mainardi

2003-12-01

Full Text Available Foi colhido um total de 442 soros em rebanhos caprinos de sete regiões do Estado de São Paulo e testados para anticorpos contra Toxoplasma gondii pela reação de imunofluorescência indireta. Em todos os rebanhos, foram encontrados caprinos reagentes, totalizando 64 (14,5% animais com sorologia positiva em diferentes capris.Four hundred forty-two serum samples were collected from dairy goats in seven regions of São Paulo State. These were tested for Toxoplasma gondii antibodies using the indirect immunofluorescent antibody test. Sixty-four (14,5% serologically positive animals were found from all these goat farms studied.

Proteomic analysis of pancreas derived from adult cloned pig

International Nuclear Information System (INIS)

Chae, Jung-Il; Cho, Young Keun; Cho, Seong-Keun; Kim, Jin-Hoi; Han, Yong-Mahn; Koo, Deog-Bon; Lee, Kyung-Kwang

2008-01-01

The potential medical applications of animal cloning include xenotransplantation, but the complex molecular cascades that control porcine organ development are not fully understood. Still, it has become apparent that organs derived from cloned pigs may be suitable for transplantation into humans. In this study, we examined the pancreas of an adult cloned pig developed through somatic cell nuclear transfer (SCNT) using two-dimensional electrophoresis (2-DE) and Western blotting. Proteomic analysis revealed 69 differentially regulated proteins, including such apoptosis-related species as annexins, lamins, and heat shock proteins, which were unanimously upregulated in the SCNT sample. Among the downregulated proteins in SCNT pancreas were peroxiredoxins and catalase. Western blot results indicate that several antioxidant enzymes and the anti-apoptotic protein were downregulated in SCNT pancreas, whereas several caspases were upregulated. Together, these data suggest that the accumulation of reactive oxygen species (ROS) in the pancreas of an adult cloned pig leads to apoptosis

Acute onset of encephalomyelitis with atypical lesions associated with dual infection of Sarcocystis neurona and Toxoplasma gondii in a dog.

Science.gov (United States)

Gerhold, Richard; Newman, Shelley J; Grunenwald, Caroline M; Crews, Amanda; Hodshon, Amy; Su, Chunlei

2014-10-15

A two-year-old male, neutered, basset hound-beagle mix with progressive neurological impairment was examined postmortem. Grossly, the dog had multiple raised masses on the spinal cord between nerve roots. Microscopically, the dog had protozoal myeloencephalitis. Toxoplasma gondii and Sarcocystis neurona were detected in the CNS by immunohistochemistry and polymerase chain reaction (PCR). Sarcocysts in formalin-fixed muscle were negative for Sarcocystis by PCR. Banked serum was negative for T. gondii using the modified agglutination test, suggesting an acute case of T. gondii infection or immunosuppression; however, no predisposing immunosuppressive diseases, including canine distemper, were found. To the authors' knowledge, this is the first report of dual T. gondii and S. neurona infection in a dog. Published by Elsevier B.V.

Molecular Characterization of Methicillin Resistant Staphylococcus aureus Strains Isolated from Intensive Care Units in Iran: ST22-SCCmec IV/t790 Emerges as the Major Clone.

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Goudarzi, Mehdi; Goudarzi, Hossein; Sá Figueiredo, Agnes Marie; Udo, Edet E; Fazeli, Maryam; Asadzadeh, Mohammad; Seyedjavadi, Sima Sadat

2016-01-01

The emergence of methicillin-resistant Staphylococcus aureus (MRSA) in different patient populations is a major public health concern. This study determined the prevalence and distribution of circulating molecular types of MRSA in hospitalized patients in ICU of hospitals in Tehran. A total of 70 MRSA isolates were collected from patients in eight hospitals. Antimicrobial resistance patterns were determined using the disk diffusion method. The presence of toxin encoding genes and the vancomycin resistance gene were determined by PCR. The MRSA isolates were further analyzed using multi-locus sequence, spa, SCCmec, and agr typing. The MRSA prevalence was 93.3%. Antimicrobial susceptibility testing revealed a high resistance rate (97.1%) to ampicillin and penicillin. The rate of resistance to the majority of antibiotics tested was 30% to 71.4%. Two isolates belonging to the ST22-SCCmec IV/t790 clone (MIC ≥ 8 μg/ml) had intermediate resistance to vancomycin. The majority of MRSA isolates (24.3%) were associated with the ST22-SCCmec IV/t790 clone; the other MRSA clones were ST859-SCCmec IV/t969 (18.6%), ST239-SCCmec III/t037 (17.1%), and ST291-SCCmec IV/t030 (8.6%). The circulating MRSA strains in Iranian hospitals were genetically diverse with a relatively high prevalence of the ST22-SCCmec IV/t790 clone. These findings support the need for future surveillance studies on MRSA to better elucidate the distribution of existing MRSA clones and detect emergence of new MRSA clones.

Detection of Toxoplasma gondii in a free-ranging giant anteater

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Thais Oliveira Morgado

Full Text Available ABSTRACT: Toxoplasmosis is caused by Toxoplasma gondii, an obligatory intracellular protozoan, which establishes acute and chronic infections in birds and mammals, including humans. This note reports, for the first time, the detection and sequencing of DNA from T. gondii in the peripheral blood of a young free range giant anteater (Myrmecophaga tridactyla. For the diagnosis, the following methods were used: polymerase chain reaction (PCR and positive serology (1:800 by means of the modified agglutination test (MAT. Since this species may be consumed by humans and predated by wild felids, its importance is emphasized as a probable source of zoonotic infection, in addition to its possible participation in the infection enzootic cycle. Although, parasitemia has been confirmed in this specimen, it presented no clinical sign of infection.

Comportamento imunológico e antigênico de cinco amostras de Toxoplasma gondii inoculadas em gatos Immunogenic and antigenic aspects from five Toxoplasma gondii strains inoculated in cats

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Italmar Teodorico Navarro

1998-09-01

Full Text Available A biologia do Toxoplasma gondii demonstra que o gato é o hospedeiro completo, responsável pela disseminação do parasito. Assim, dois gatos domésticos foram imunizados com cada uma das amostras, VPS (humano, LIV-IV e LIV-V (suíno, CPL (caprino e CN (felino de T. gondii. Foram utilizados taquizoítas vivos em inóculos endovenosos de 2x10(6 (1° inóculo} e 4x10(7 (2° inóculo - 35 dias após, exceto a amostra VPS, onde 1 gato morreu no 10° dia com sinais clínicos agudos da doença, nas outras amostras, nenhum sinal clínico foi constatado durante os 6 meses de observação. O nível de anticorpos na imunização foi acompanhado através da reação de imunofluorescência indireta (IFI com conjugado anti-IgG de felino. Os títulos de anticorpos obtidos no 20° dia variaram de 1:1.024 a 1:4.096 e de 1:1.024 a 1:8.000 no 40° dia. Somente a amostra VPS expressou títulos de 1:16.000 no 30° dia da imunização. Títulos homólogos e heterólogos foram equivalentes sem nenhuma diferença entre as amostras. Quando soros imunes foram adsorvidos com taquizoítas vivos de cada amostra, a redução nos títulos de anticorpos foi demonstrada em ambos homólogos e heterólogos. Esses resultados sugerem que, embora diferente em virulência para gatos, a superfície antigênica é comum entre as amostras do T. gondii, com base no nível de anticorpo demonstrado pela IFI. Esses resultados também demonstram que, aparentemente, não há correlação entre virulência e as características sorológicas das amostras estudadas no T. gondii. Entretanto, a importância do teste IFI em diagnóstico laboratorial é reforçada.The biology of Toxoplasma gondii demonstrate that cats are the complete host responsable for the dissemination of this parasites. Two domestic cats were immunised with Toxoplasma gondii strains VPS (human, LIV-IV and LIV-V (porcine, CPL (caprine and CN (feline. Live tachyzoites were utilized in intravenous inoculation of 2x10(7 (first

Seroprevalence of antibody to TgGRA7 antigen of Toxoplasma gondii in livestock animals from Western Java, Indonesia.

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Ichikawa-Seki, Madoka; Guswanto, Azirwan; Allamanda, Puttik; Mariamah, Euis Siti; Wibowo, Putut Eko; Igarashi, Ikuo; Nishikawa, Yoshifumi