Comparison of six simple methods for extracting ribosomal and mitochondrial DNA from Toxocara and Toxascaris nematodes.

Abstract:

:Six simple methods for extraction of ribosomal and mitochondrial DNA from Toxocara canis, Toxocara cati and Toxascaris leonina were compared by evaluating the presence, appearance and intensity of PCR products visualized on agarose gels and amplified from DNA extracted by each of the methods. For each species, two isolates were obtained from the intestines of their respective hosts: T. canis and T. leonina from dogs, and T. cati from cats. For all isolates, total DNA was extracted using six different methods, including grinding, boiling, crushing, beating, freeze-thawing and the use of a commercial kit. To evaluate the efficacy of each method, the internal transcribed spacer (ITS) region and the cytochrome c oxidase subunit 1 (cox1) gene were chosen as representative markers for ribosomal and mitochondrial DNA, respectively. Among the six DNA extraction methods, the beating method was the most cost effective for all three species, followed by the commercial kit. Both methods produced high intensity bands on agarose gels and were characterized by no or minimal smear formation, depending on gene target; however, beating was less expensive. We therefore recommend the beating method for studies where costs need to be kept at low levels.

journal_name

Exp Parasitol

authors

Mikaeili F,Kia EB,Sharbatkhori M,Sharifdini M,Jalalizand N,Heidari Z,Zarei Z,Stensvold CR,Mirhendi H

doi

10.1016/j.exppara.2013.02.008

subject

Has Abstract

pub_date

2013-06-01 00:00:00

pages

155-9

issue

2

eissn

0014-4894

issn

1090-2449

pii

S0014-4894(13)00050-7

journal_volume

134

pub_type

杂志文章
  • Trichomonas vaginalis: characterization of its glutamate dehydrogenase.

    abstract::An NADP-linked glutamate dehydrogenase (EC 1.4.1.4) was found in the soluble fraction of Trichomonas vaginalis. Its molecular weight was about 230,000 (gel filtration). The enzyme, partially purified by diafiltration and hydroxyapatite column chromatography, was heat stable (1 hr at 57 C). It catalyzed both the aminat...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(88)90007-0

    authors: Turner AC,Lushbaugh WB

    更新日期:1988-10-01 00:00:00

  • Trypanosoma cruzi (Kinetoplastida Trypanosomatidae): ecology of the transmission cycle in the wild environment of the Andean valley of Cochabamba, Bolivia.

    abstract::An active Trypanosoma cruzi transmission cycle maintained by wild rodents in the Andean valleys of Cochabamba Bolivia is described. Wild and domestic Triatoma infestans with 60% infection with T. cruzi were found and was evidenced in 47.5% (rodents) and 26.7% (marsupial) by parasitological and/or serologycal methods. ...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2006.04.010

    authors: Cortez MR,Pinho AP,Cuervo P,Alfaro F,Solano M,Xavier SC,D'Andrea PS,Fernandes O,Torrico F,Noireau F,Jansen AM

    更新日期:2006-12-01 00:00:00

  • Giardia intestinalis: conservation of the variant-specific surface protein VSP417-1 (TSA417) and identification of a divergent homologue encoded at a duplicated locus in genetic group II isolates.

    abstract::The stability of the gene encoding TSA417, a 72-kDa variant-specific surface protein (VSP) produced by trophozoites of Giardia intestinalis isolate WB-C6, was investigated in isolates of similar (Assemblage A / Group I) or distinct (Assemblage A / Group II) genotype. Using primers specific for the WB-C6 tsa417 gene, D...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1006/expr.1998.4325

    authors: Ey PL,Darby JM

    更新日期:1998-11-01 00:00:00

  • Prodigiosin is not a determinant factor in lysis of Leishmania (Viannia) braziliensis after interaction with Serratia marcescens D-mannose sensitive fimbriae.

    abstract::In this paper, the lytic activity of two variants of Serratia marcescens against promastigotes of Leishmania braziliensis was studied. In vitro assays showed that S. marcescens variant SM365 lyses L. braziliensis promastigotes, while the variant DB11 did not. Scanning electron microscopy (SEM) revealed that S. marcesc...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2009.03.004

    authors: Moraes CS,Seabra SH,Albuquerque-Cunha JM,Castro DP,Genta FA,de Souza W,Brazil RP,Garcia ES,Azambuja P

    更新日期:2009-06-01 00:00:00

  • Trypanosoma cruzi: quantification in tissues of experimentally infected mice by limiting dilution analysis.

    abstract::A limiting dilution assay (LDA) was developed for the quantification of Trypanosoma cruzi in the heart and blood of infected mice. Three groups of swiss mice were injected ip with "CL", "Colombiana," and "Y" strains. At 1-day intervals after infection, blood and the heart were removed. Serial blood dilutions in LIT me...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(90)90099-x

    authors: Nunes MP,Coutinho SG,Louis JA,Souza WJ

    更新日期:1990-02-01 00:00:00

  • BmSI-7, a novel subtilisin inhibitor from Boophilus microplus, with activity toward Pr1 proteases from the fungus Metarhizium anisopliae.

    abstract::BmSI-7 and BmSI-6, two Boophilus microplus subtilisin inhibitors (BmSI) were purified and characterized from eggs. The inhibitors isolated by classical purification methods presented molecular masses of 7408 and 7271Da, respectively, by MALDI-TOF-MS. Both BmSI-7 and BmSI-6 inhibited neutrophil elastase (K(i) 0.4 and 0...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2007.08.003

    authors: Sasaki SD,de Lima CA,Lovato DV,Juliano MA,Torquato RJ,Tanaka AS

    更新日期:2008-02-01 00:00:00

  • Plasmodium falciparum: food vacuole localization of nitric oxide-derived species in intraerythrocytic stages of the malaria parasite.

    abstract::Nitric oxide (NO) has diverse biological functions. Numerous studies have documented NO's biosynthetic pathway in a wide variety of organisms. Little is known, however, about NO production in intraerythrocytic Plasmodium falciparum. Using diaminorhodamine-4-methyl acetoxymethylester (DAR-4M AM), a fluorescent indicato...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2008.04.014

    authors: Ostera G,Tokumasu F,Oliveira F,Sa J,Furuya T,Teixeira C,Dvorak J

    更新日期:2008-09-01 00:00:00

  • Homologues of the 24-kDa flagellar Ca(2+)-binding protein gene of Trypanosoma cruzi are present in other members of the Trypanosomatidae family.

    abstract::A full-length cDNA encoding the 24-kDa flagellar Ca(2+)-binding protein (FCaBP) of the Dm28c clone of Trypanosoma cruzi was cloned and characterized. Comparison of the deduced amino acid sequence with those of the FCaBPs of other T. cruzi strains revealed greater than 97% sequence conservation. FCaBP-like genes are fo...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1006/expr.1997.4159

    authors: Maldonado RA,Linss J,Thomaz N,Olson CL,Engman DM,Goldenberg S

    更新日期:1997-07-01 00:00:00

  • Eimeria tenella: characterization of a 5S ribosomal RNA repeat unit and its use as a species-specific probe.

    abstract::The oocysts of Eimeria tenella, one of the most pathogenic of several species causing chicken coccidiosis, are difficult to distinguish microscopically from several other infective Eimeria species. One copy of a gene coding for 5S ribosomal RNA has been cloned from E. tenella and sequenced. A coding region of 120 nucl...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1006/expr.1993.1008

    authors: Stucki U,Braun R,Roditi I

    更新日期:1993-02-01 00:00:00

  • The time course of selected malarial infections in cytokine-deficient mice.

    abstract::Murine malarial parasites have long been characterized by their requirement for either antibody-mediated immunity (AMI) or cell-mediated immunity (CMI) for suppression of acute parasitemia, with Plasmodium yoelii reportedly requiring AMI for suppression and P. chabaudi requiring CMI. To assess this characterization in...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1006/expr.1996.4132

    authors: van der Heyde HC,Pepper B,Batchelder J,Cigel F,Weidanz WP

    更新日期:1997-02-01 00:00:00

  • Schistosoma mansoni: influence of infection on mouse behavior.

    abstract::Schistosoma mansoni infection in humans and animals induces abnormal neurobehavioral responses following granuloma formation. In mice, granulomas in the liver are observed 8 weeks after infection, while after 15-20 weeks, the presence of eggs and granulomas in the brain has been reported. In this study, outbred CD-1 f...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1006/expr.1996.0047

    authors: Fiore M,Moroni R,Alleva E,Aloe L

    更新日期:1996-06-01 00:00:00

  • Screening and characterization of apical membrane antigen 1 interacting proteins in Eimeria tenella.

    abstract::Avian coccidiosis is a widespread and economically significant disease of poultry. It is an enteric disease caused by several protozoan Eimeria species. Eimeria belongs to the phylum Apicomplexa, which exhibits an unusual mechanism of host cell invasion. During invasion of host cells, the protein apical membrane antig...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2016.09.014

    authors: Han H,Xue P,Dong H,Zhu S,Zhao Q,Huang B

    更新日期:2016-11-01 00:00:00

  • Excretory/secretory products from the gastrointestinal nematode Trichuris muris.

    abstract::To better control gastrointestinal nematode infections in humans and animals, it is important to understand the strategies used by these parasites to modulate the host immune system. In this regard, molecules released by parasites have been attributed crucially important roles in host-parasite negotiations. We charact...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2017.05.003

    authors: Tritten L,Tam M,Vargas M,Jardim A,Stevenson MM,Keiser J,Geary TG

    更新日期:2017-07-01 00:00:00

  • Behavioral and hormonal changes associated with the infective dose in experimental taeniasis in golden hamsters (Mesocricetus auratus).

    abstract:INTRODUCTION:It has been reported that behavioral changes relate to infection in different parasitoses. However, the relation between the extent of the behavioral changes and the magnitude of the infection has been scarcely studied. The aim of this study was to evaluate the relationship between different doses of infec...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2016.04.013

    authors: Domínguez-Roldan R,Hallal-Calleros C,Sciutto E,Hernández M,Aguirre-Flores V,García-Jiménez S,Báez-Saldaña A,Flores-Pérez FI

    更新日期:2016-07-01 00:00:00

  • Leishmania braziliensis: localization of glycoproteins in promastigotes.

    abstract::Two species of glycoproteins from Leishmania braziliensis promastigotes of apparent molecular weights of 53,000 (glycoprotein 53) and 47,000 (glycoprotein 47) were localized. Four lectins with different sugar specificities bound to the blotting sheet to which the electrophoretically separated materials were transferre...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(86)90188-8

    authors: Nagakura K,Tachibana H,Kaneda Y,Nakae T

    更新日期:1986-06-01 00:00:00

  • Toxoplasma gondii: P30 peptides recognition pattern in human toxoplasmosis.

    abstract::In this study, human sera reactivity against nine peptides derived from the Toxoplasma gondii P30 protein was assessed by ELISA in patients with different clinical forms of toxoplasmosis. Same as has been reported in mice, sera from congenital, ocular and chronic asymptomatic toxoplasmosis patients recognized more str...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2009.06.017

    authors: Cardona N,de-la-Torre A,Siachoque H,Patarroyo MA,Gomez-Marin JE

    更新日期:2009-10-01 00:00:00

  • Behavioral responses of Poecilia vivipara (Osteichthyies: Cyprinodontiformes) to experimental infections of Acanthocollaritrema umbilicatum (Digenea: Cryptogonimidae).

    abstract::The swimming behavior of Poecilia vivipara was evaluated using an image analysis system comparing laboratory-reared uninfected fish before and after experimental infection with different intensities of cercariae of the trematode Acanthocollaritrema umbilicatum. Two experiments were performed, each with 30 fish which w...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2010.10.018

    authors: Santos EG,Cunha RA,Santos CP

    更新日期:2011-02-01 00:00:00

  • Acanthamoeba spp.: in vitro effects of clinical isolates on murine macrophages, osteosarcoma and HeLa cells.

    abstract::Three different cell lines (murine macrophages, HeLa and osteosarcoma cells) were assayed in order to check for the manifestation of the cytopathic effects of three strains of Acanthamoeba recently isolated in our laboratory from contact lens cases: CLC-16, CLC-41.r and CLC-51-l. Adhesion and cytotoxicity assays were ...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2009.10.011

    authors: Martín-Navarro CM,Lorenzo-Morales J,Machín RP,López-Arencibia A,Valladares B,Piñero JE

    更新日期:2010-09-01 00:00:00

  • Genotyping, physiological features and proteolytic activities of a potentially pathogenic Acanthamoeba sp. isolated from tap water in Brazil.

    abstract::Acanthamoeba spp., known to cause keratitis and granulomatous encephalitis in humans, are frequently isolated from a variety of water sources. Here we report for the first time the characterization of an Acanthamoeba sp. (ACC01) isolated from tap water in Brazil. This organism is currently being maintained in an axeni...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2009.07.006

    authors: Magliano AC,da Silva FM,Teixeira MM,Alfieri SC

    更新日期:2009-11-01 00:00:00

  • Toxoplasma gondii: effects of Artemisia annua L. on susceptibility to infection in experimental models in vitro and in vivo.

    abstract::Considering that the treatment for toxoplasmosis is based on drugs that show limited efficacy due to their substantial side effects, the purpose of the present study was to evaluate the effects of Artemisia annua on in vitro and in vivo Toxoplasma gondii infection. A. annua infusion was prepared from dried herb and te...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2009.04.010

    authors: de Oliveira TC,Silva DA,Rostkowska C,Béla SR,Ferro EA,Magalhães PM,Mineo JR

    更新日期:2009-07-01 00:00:00

  • Toxoplasma gondii isolates: multilocus RFLP-PCR genotyping from human patients in Sao Paulo State, Brazil identified distinct genotypes.

    abstract::This study investigated the genetic characteristics of Toxoplasma gondii samples collected from 62 patients with toxoplasmosis in Sao Paulo State, Brazil. DNA samples were isolated from blood, cerebrospinal fluid and amniotic fluids of 25 patients with cerebral toxoplasmosis and AIDS, two patients with acute toxoplasm...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2011.06.002

    authors: Ferreira IM,Vidal JE,de Mattos Cde C,de Mattos LC,Qu D,Su C,Pereira-Chioccola VL

    更新日期:2011-10-01 00:00:00

  • Arginase activity is associated with fibrosis in experimental infection with Taenia crassiceps, but does not play a major role in resistance to infection.

    abstract::Murine infection with Taenia crassiceps cysticerci is used as an experimental model for human and animal cysticercosis. In this infection parasites can be found associated with an inflammatory infiltrate enriched with macrophages. Experimental evidence exists supporting a role for either NO-producing classically activ...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2013.09.014

    authors: Moura VB,Silva MM,Batista LF,Gomes CM,Leenen PJ,Lino RS Jr,Oliveira MA

    更新日期:2013-11-01 00:00:00

  • Molecular mechanisms of host cell invasion by Trypanosoma cruzi.

    abstract::The protozoan parasite Trypanosoma cruzi, the etiologic agent of Chagas disease, is an obligate intracellular protozoan pathogen. Overlapping mechanisms ensure successful infection, yet the relationship between these cellular events and clinical disease remains obscure. This review explores the process of cell invasio...

    journal_title:Experimental parasitology

    pub_type: 杂志文章,评审

    doi:10.1016/j.exppara.2010.06.023

    authors: Epting CL,Coates BM,Engman DM

    更新日期:2010-11-01 00:00:00

  • Plasmodium falciparum and Plasmodium berghei: effect of magnesium on the development of parasitemia.

    abstract::The in vitro growth of Plasmodium falciparum was reduced by 35 and 43% through high concentrations (5 mmole/liter) of magnesium in RPMI medium and magnesium-free medium, respectively, after 48 hr, whereas no significant inhibition could be observed under these conditions after 24 hr cultivation in the respective mediu...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1006/expr.1995.1023

    authors: Hess FI,Kilian A,Söllner W,Nothdurft HD,Pröll S,Löscher T

    更新日期:1995-03-01 00:00:00

  • Molecular cloning and characterization of leucine aminopeptidase from Fasciola gigantica.

    abstract::M17 leucine aminopeptidase (LAP) is one of a family of metalloexopeptidases, of which short peptide fragments are cleaved from the N-terminals. In this study, the full length of cDNA encoding Fasciola gigantica LAP (FgLAP) was cloned from adult parasites. The amino acid sequences of FgLAP showed a high degree of ident...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2012.04.008

    authors: Changklungmoa N,Chaithirayanon K,Kueakhai P,Meemon K,Riengrojpitak S,Sobhon P

    更新日期:2012-07-01 00:00:00

  • Babesia gibsoni: identification of an immunodominant, interspersed repeat antigen.

    abstract::In this report, an immunodominant antigen called BgIRA from Babesia gibsoni is identified and described. A highly repetitive antigen was screened from a cDNA library. The genomic BgIRA gene exists as single cope gene and contains 10 introns. BgIRA plays a dominant role in the immune response in dogs infected with B. g...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2007.07.005

    authors: Jia H,Terkawi MA,Aboge GO,Goo YK,Zhou J,Lee EG,Nishikawa Y,Igarashi I,Fujisaki K,Xuan X

    更新日期:2008-01-01 00:00:00

  • Acanthamoeba castellanii: in vitro effects of selected biological, physical and chemical factors.

    abstract::Trophozoites and cysts of free-living Acanthamoeba castellanii present a serious risk to human health as causative agents of human diseases such as fatal granulomatous amoebic encephalitis and Acanthamoeba keratitis that is reported from various part of the world, also in Poland, with increasing frequency, particularl...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2010.01.025

    authors: Chomicz L,Padzik M,Graczyk Z,Starosciak B,Graczyk TK,Naprawska A,Oledzka G,Szostakowska B

    更新日期:2010-09-01 00:00:00

  • Cloning, overexpression and characterization of Leishmania donovani triosephosphate isomerase.

    abstract::Triosephosphate isomerase (TIM) is a major enzyme in the glycolytic pathway, which catalyzes the interconversion of glyceraldehyde 3-phosphate to dihydroxyacetone phosphate. Here, we report cloning, expression and purification of a catalytically active recombinant TIM of Leishmania donovani (LdTIM). The recombinant Ld...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2012.01.016

    authors: Kumar K,Bhargava P,Roy U

    更新日期:2012-04-01 00:00:00

  • Trypanosoma cruzi: Oxidative stress induces arginine kinase expression.

    abstract::Trypanosoma cruzi arginine kinase is a key enzyme in cell energy management and is also involved in pH and nutritional stress response mechanisms. T. cruzi epimastigotes treated with hydrogen peroxide presented a time-dependent increase in arginine kinase expression, up to 10-fold, when compared with untreated parasit...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/j.exppara.2006.04.004

    authors: Miranda MR,Canepa GE,Bouvier LA,Pereira CA

    更新日期:2006-12-01 00:00:00

  • Fasciola hepatica and Schistosoma mansoni: immunofluorescent antigen localization and cross-reactivity.

    abstract::In an attempt to identify the tissue sources of biochemically purified antigenic fractions of Fasciola hepatica and Schistosoma mansoni, antisera were tested against plastic-embedded sections of worms of various ages by an indirect fluorescent-antibody-labeling technique. Antibodies prepared against antigens purified ...

    journal_title:Experimental parasitology

    pub_type: 杂志文章

    doi:10.1016/0014-4894(84)90055-9

    authors: Hanna RE,Hillyer GV

    更新日期:1984-02-01 00:00:00