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1.
J Water Health ; 16(1): 138-149, 2018 Feb.
Article in English | MEDLINE | ID: mdl-29424727

ABSTRACT

Water resources situated in areas with underlying karst geology are particularly vulnerable to fecal pollution. In such vulnerable systems, microbial source tracking (MST) methods are useful tools to elucidate the pathways of both animal and human fecal pollution, leading to more accurate water use risk assessments. Here, we describe the application of a MST toolbox using both culture-dependent bacteriophage and molecular-dependent 16S rRNA assays at spring and well sites in the karstic St Imier Valley, Switzerland. Culture-dependent and molecular-dependent marker performance varied significantly, with the 16S rRNA assays displaying greater sensitivity than their phage counterpart; HF183 was the best performing human wastewater-associated marker while Rum2Bac was the best performing ruminant marker. Differences were observed in pollution regimes between the well and spring sampling sites, with the spring water being more degraded than the well site. Our results inform the choice of marker selection for MST studies and highlight differences in microbial water quality between well and spring karst sites.


Subject(s)
Drinking Water/microbiology , Environmental Monitoring/methods , Water Microbiology , Water Resources , Animals , Bacteriophages/isolation & purification , Humans , Polymerase Chain Reaction , Risk Assessment , Switzerland , Viruses/isolation & purification , Water Quality
2.
Environ Sci Technol ; 49(12): 7142-51, 2015 Jun 16.
Article in English | MEDLINE | ID: mdl-25871525

ABSTRACT

Microbial contamination of groundwater represents a significant health risk to resource users. Culture-dependent Bacteroides phage and molecular-dependent Bacteroidales 16S rRNA assays are employed in microbial source tracking (MST) studies globally, however little is known regarding how these important groups relate to each other in the environment and which is more suitable to indicate the presence of waterborne fecal pollution and human enteric viruses. This study addresses this knowledge gap by examining 64 groundwater samples from sites with varying hydrogeological properties using a MST toolbox containing two bacteriophage groups (phage infecting GB-124 and ARABA-84), and two Bacteroidales 16S rRNA markers (Hf183 and BacR); those were compared to fecal indicator bacteria, somatic coliphage, Bacteroidales 16S rRNA marker AllBac, four human enteric viruses (norovirus GI and II, enterovirus and group A rotavirus) and supplementary hydrogeological/chemical data. Bacteroidales 16S rRNA indicators offered a more sensitive assessment of both human-specific and general fecal contamination than phage indicators, but may overestimate the risk from enteric viral pathogens. Comparison with hydrogeological and land use site characteristics as well as auxiliary microbiological and chemical data proved the plausibility of the MST findings. Sites representing karst aquifers were of significantly worse microbial quality than those with unconsolidated or fissured aquifers, highlighting the vulnerability of these hydrogeological settings.


Subject(s)
Bacteroides/genetics , Bacteroidetes/genetics , Biomarkers/analysis , Feces/virology , Groundwater/virology , Water Pollution/analysis , Water Quality , Feces/microbiology , Groundwater/microbiology , Groundwater/standards , Humans , RNA, Ribosomal, 16S/genetics
3.
Parasitology ; 122(Pt 2): 125-32, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11272643

ABSTRACT

The cloning and characterization of Ts-p41, an EF-hand calcium-binding protein of the protozoan parasite Tritrichomonas suis is described. A T. suis cDNA library was screened with monospecific antibodies affinity purified on an immunoreactive 41 kDa antigen in a Triton X-114 membrane-protein fraction. The resulting cDNA fragments turned out to be derived from 2 different genes encoding closely related Ts-p41 variants. The deduced amino acid sequences contained 6 EF-hand domains perfectly matching the canonical consensus motif and a putative C-terminal prenylation site. Northern and Southern hybridizations revealed that Ts-p41 was highly expressed and encoded by a gene-family. A cDNA encoding Ts-p41 was expressed as recombinant protein in Escherichia coli. By overlay with 45Ca it was demonstrated that the native and recombinant Ts-p41 proteins bind Ca2+. In immunofluorescence, epitopes recognized by anti-Ts-p41 antibodies were distributed as well on the anterior flagella as on the recurrent flagellum of the parasite. Our findings with the parabasalid T. suis suggest that multiple EF-hand bearing calcium-binding proteins might be a common phenomenon associated with flagellar motility.


Subject(s)
Calcium-Binding Proteins/chemistry , Calcium-Binding Proteins/genetics , Tritrichomonas/chemistry , Amino Acid Sequence , Animals , Calcium/metabolism , Calcium-Binding Proteins/metabolism , Cloning, Molecular , Electrophoresis, Polyacrylamide Gel , Flagella/chemistry , Fluorescent Antibody Technique , Molecular Sequence Data , Random Amplified Polymorphic DNA Technique , Recombinant Fusion Proteins/chemistry , Tritrichomonas/genetics
4.
Mol Biochem Parasitol ; 108(1): 109-17, 2000 Apr 30.
Article in English | MEDLINE | ID: mdl-10802323

ABSTRACT

The cloning and characterization of Ts-p66, a calcium-binding protein representing calnexin of the protozoan parasite Tritrichomonas suis is described. A T. suis cDNA expression library was screened with monospecific antibodies affinity-purified on an immuno-reactive 66 kDa antigen in a Triton X-114 membrane-protein fraction. The deduced amino acid sequence of the resulting cDNA clones revealed that Ts-p66 belongs to the calreticulin protein family and represents calnexin of T. suis. The key structural features and sequence motifs of the calnexins were all conserved. By lectin-blotting we demonstrated that the native protein is glycosylated. Northern and Southern hybridizations showed that T. suis calnexin was highly expressed and encoded by a single or low copy number gene. A cDNA encoding Ts-p66 was expressed as recombinant protein in Escherichia coli. By overlay with 45Ca it was demonstrated that the native and recombinant proteins bind Ca(2+). Using immunofluorescence with affinity-purified antibodies, a staining pattern was observed which points towards a putative localization of Ts-p66 in the nuclear membrane and endoplasmic reticulum. Demonstration of a structurally conserved calnexin in the amitochondriate protist T. suis indicates the very early evolutionary origin of the machinery for quality control of protein folding in the endoplasmic reticulum and the molecules involved hereby.


Subject(s)
Calcium-Binding Proteins/genetics , Calcium-Binding Proteins/metabolism , Conserved Sequence , Tritrichomonas/genetics , Tritrichomonas/metabolism , Amino Acid Motifs , Amino Acid Sequence , Animals , Calcium/metabolism , Calcium-Binding Proteins/chemistry , Calnexin , Cloning, Molecular , DNA, Complementary , Endoplasmic Reticulum/metabolism , Evolution, Molecular , Fluorescent Antibody Technique , Molecular Sequence Data , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Sequence Analysis, DNA
5.
Parasitol Res ; 85(3): 246-8, 1999 Mar.
Article in English | MEDLINE | ID: mdl-9951970

ABSTRACT

Reverse-transcriptase polymerase chain reaction cloning of the 3'-ends of the alpha-tubulin cDNAs of Tritrichomonas mobilensis in combination with Southern-blot analysis identified seven to eight distinct alpha-tubulin genes. All seven lack a carboxy-terminal tyrosine and the corresponding sequence compatible with posttranslational tyrosination. This indicates that whereas tyrosination of alpha-tubulin has been found in most species, including humans and trypanosomes, it is absent in trichomonads.


Subject(s)
Genes, Protozoan , Multigene Family , Tritrichomonas/genetics , Tubulin/genetics , Tyrosine , Amino Acid Sequence , Animals , Base Sequence , Blotting, Southern , Cloning, Molecular , DNA, Protozoan/genetics , Humans , Molecular Sequence Data , Protein Processing, Post-Translational , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Tubulin/chemistry
6.
Microbes Infect ; 1(10): 807-16, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10816086

ABSTRACT

Tritrichomonas foetus is a parasite of particular veterinary importance causing bovine tritrichomonosis, a sexually transmitted disease leading to infertility and abortion. The present review summarizes the current knowledge on potential mechanisms of pathogenicity of T. foetus, the immunology of host-parasite interaction in bovine tritrichomonosis, and the experimental model systems of this parasitic disease.


Subject(s)
Cattle Diseases/parasitology , Protozoan Infections, Animal , Tritrichomonas foetus/parasitology , Abortion, Veterinary/immunology , Abortion, Veterinary/parasitology , Abortion, Veterinary/pathology , Animals , Cattle , Cattle Diseases/immunology , Cattle Diseases/pathology , Female , Host-Parasite Interactions/immunology , Mice , Pregnancy , Protozoan Infections/pathology , Tritrichomonas foetus/immunology
7.
Parasite Immunol ; 20(10): 473-81, 1998 Oct.
Article in English | MEDLINE | ID: mdl-9797508

ABSTRACT

By a strategy of differential immunological screening of an expression library constructed from adult Echinococcus multilocularis parasites, a partial cDNA sequence encoding a protein termed Em6 was isolated. This molecule displayed high sequence homology to the recombinant antigen 'Eg6' which was previously described as an immunogenic epitope of antigen 5 of E. granulosus. Further Em6 sequences and the corresponding sequences from a cattle isolate of E. granulosus were obtained by a PCR approach. By immunoblot analyses using affinity purified antibodies, expression of Em6 in fertile cysts producing protoscoleces of the E. multilocularis metacestode stage was observed. However, Em6 was absent in non-fertile metacestodes. The demonstration of a protein in E. multilocularis displaying identities to 'antigen 6' of E. granulosus could potentially contribute to the future elucidation of the relationship between antigen 5 and 'antigen 6' in the genus Echinococcus, and shed some lights on the performance of serodiagnostic assays for hydatid disease based on the respective antigens.


Subject(s)
Antigens, Helminth/immunology , Echinococcus/chemistry , Helminth Proteins/genetics , Helminth Proteins/immunology , Amino Acid Sequence , Animals , Antibodies, Helminth/blood , Base Sequence , Blotting, Western , Cattle , DNA, Complementary , Dogs , Echinococcus/immunology , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Epitopes/immunology , Gerbillinae , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Polymerase Chain Reaction , Rabbits , Sequence Analysis , Sequence Homology, Amino Acid
8.
FEBS Lett ; 429(3): 399-402, 1998 Jun 16.
Article in English | MEDLINE | ID: mdl-9662457

ABSTRACT

The alpha- and beta-tubulins present in cytoskeletons of Tritrichomonas mobilensis are extensively glutamylated. Automated sequencing and mass spectrometry of the carboxyterminal peptides identifies 4 glutamylation sites in alpha- and 2 sites in beta-tubulin. They are marked by asterisks in the terminal sequences GDE*E*E*E*DDG (alpha) and EGE*E*DEEAEA (beta). This is the first report that tubulin glutamylation can occur at multiple sites. Although T. mobilensis has four flagellae the tubulins lack polyglycylation. Thus glycylation is not necessary for formation or function of axonemal microtubules. Alpha-tubulin is completely acetylated at lysine 40 and shows no tyrosine cycle. Peptide sequences establish two distinct beta-tubulins.


Subject(s)
Glutamic Acid/metabolism , Protein Processing, Post-Translational , Trichomonas/metabolism , Tubulin/metabolism , Amino Acid Sequence , Animals , Cytoskeleton/metabolism , Eukaryotic Cells/metabolism , Mass Spectrometry , Molecular Sequence Data , Peptide Fragments/chemistry , Sequence Analysis , Tubulin/chemistry
9.
Mol Biochem Parasitol ; 91(2): 281-93, 1998 Mar 15.
Article in English | MEDLINE | ID: mdl-9566521

ABSTRACT

A cDNA expression library representing the metacestode developmental stage of the tapeworm Echinococcus multilocularis was immunoscreened with monospecific antibodies affinity purified following differential immunoblot analysis. Using this procedure, a metacestode-specific clone was isolated representing a 14-3-3 gene of the parasite, which is present as a single copy in the parasite genome. The identity of this clone was demonstrated by cross-reactivity of the recombinant E. multilocularis 14-3-3 protein with antibodies raised against a heterologous 14-3-3 protein from Saccharomyces cerevisiae. In addition, expression of the E. multilocularis 14-3-3 gene in the mutant S. cerevisiae strain, DS9-22, resulted in complementation of the phenotypic deficiency of this strain, thus demonstrating the functionality of the respective gene product. By reverse transcription-polymerase chain reaction (RT-PCR) we showed that the E. multilocularis 14-3-3 protein is about 10-fold overexpressed in the metacestode stage compared with the expression level in the adult stage. Immunolocalization of the 14-3-3 protein in E. multilocularis metacestodes revealed its predominant presence in the germinal layer of the parasite. The results of this study, taken together with the current knowledge on the 14-3-3 protein family, suggest that this parasite molecule may contribute to the promotion of the progressive, potentially unlimited growth behaviour of the E. multilocularis metacestode within the host tissue.


Subject(s)
Echinococcus/genetics , Gene Expression Regulation, Developmental , Genes, Helminth , Helminth Proteins/genetics , Proteins/genetics , Tyrosine 3-Monooxygenase , 14-3-3 Proteins , Amino Acid Sequence , Animals , Antigens, Helminth/chemistry , Antigens, Helminth/genetics , Antigens, Helminth/immunology , Cross Reactions , Echinococcus/chemistry , Echinococcus/growth & development , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique , Genetic Complementation Test , Helminth Proteins/chemistry , Helminth Proteins/immunology , Humans , Immunoblotting , Life Cycle Stages , Mice , Molecular Probe Techniques , Molecular Sequence Data , Polymerase Chain Reaction , Proteins/chemistry , Proteins/immunology , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/growth & development
10.
Parasitol Res ; 84(2): 153-6, 1998.
Article in English | MEDLINE | ID: mdl-9493217

ABSTRACT

The taxonomic classification within the genus Tritrichomonas is a subject of controversy, and, potentially, separation of the tritrichomonads from cattle and swine on the species level is not valid. To tackle this hypothesis we comparatively assessed several isolates of protozoan parasites from the three Tritrichomonas species T. foetus, T. suis, and T. mobilensis by the RAPD (random amplified polymorphic DNA) technique. In this method with 20 different primers, all T. foetus and T. suis isolates resulted in identical genomic fingerprints, thus yielding additional evidence for the genetic identity of T. foetus and T. suis. In contrast, it turned out that the species T. mobilensis isolated from the squirrel monkey is genetically distinct and can clearly be discriminated from the other tritrichomonads. Consequently, the results obtained in this study support a possible future revision of the taxonomic classification of the genus Tritrichomonas.


Subject(s)
DNA Fingerprinting , Tritrichomonas/classification , Tritrichomonas/genetics , Animals , Cattle , DNA, Protozoan/genetics , Random Amplified Polymorphic DNA Technique , Saimiri , Species Specificity , Swine
11.
J Clin Microbiol ; 36(2): 513-9, 1998 Feb.
Article in English | MEDLINE | ID: mdl-9466768

ABSTRACT

Tritrichomonas foetus is the causative agent of bovine tritrichomonosis, a sexually transmitted disease leading to infertility and abortion. Diagnosis is hampered by putative contamination of samples with intestinal or coprophilic trichomonadid protozoa which might be mistaken for T. foetus. Therefore, we developed a PCR test optimized for applicability in routine diagnosis. Amplification is based upon primers TFR3 and TFR4 directed to the rRNA gene units of T. foetus. In order to avoid potential carryover contamination by products of previous amplification reactions, conditions were adapted to the use of the uracil DNA glycosylase system. Furthermore, documentation and interpretation of results were facilitated by including a DNA enzyme immunoassay for the detection of amplification products. Specificity was confirmed with genomic material from different related trichomonadid protozoa. The high sensitivity of the test allowed the detection of a single T. foetus organism in diagnostic culture medium or about 50 parasites per ml of preputial washing fluid. The present methods are thus proposed as (i) confirmatory tests for microscopic diagnosis following diagnostic in vitro cultivation and (ii) a direct T. foetus screening test with diagnostic samples.


Subject(s)
DNA, Protozoan/isolation & purification , N-Glycosyl Hydrolases/immunology , Polymerase Chain Reaction/methods , Protozoan Infections/diagnosis , Tritrichomonas foetus/isolation & purification , Animals , Cattle , Culture Media , DNA Glycosylases , DNA Primers/genetics , DNA, Protozoan/analysis , Diagnosis, Differential , Female , Genome, Protozoan , Immunoenzyme Techniques , Male , Protozoan Infections, Animal , RNA, Ribosomal/genetics , RNA, Ribosomal/metabolism , Sensitivity and Specificity , Trichomonas/genetics , Trichomonas/immunology , Trichomonas/isolation & purification , Tritrichomonas/genetics , Tritrichomonas/immunology , Tritrichomonas/isolation & purification , Tritrichomonas foetus/genetics , Tritrichomonas foetus/immunology
12.
Parasitology ; 115 ( Pt 2): 111-9, 1997 Aug.
Article in English | MEDLINE | ID: mdl-10190167

ABSTRACT

The taxonomic situation in the genus Tritrichomonas is the subject of controversial discussion: potentially T. foetus and T. suis, the tritrichomonads from cattle and swine, respectively, could belong to the same species. In order to shed some light on this question, a molecular biological analysis was performed. The 5.8S rRNA gene and the flanking internal transcribed spacer regions (ITS1 and ITS2) of 12 different isolates of 3 Tritrichomonas species T. foetus, T. suis and T. mobilensis were enzymatically amplified by PCR and subcloned. Also, the corresponding regions of the trichomonads Trichomonas vaginalis, T. tenax, T. gallinae and Pentatrichomonas hominis were included in this study. Sequence analysis of cloned fragments was used to compare the parasite isolates. The genus Tritrichomonas exhibited an extremely high degree of homogeneity. All T. foetus and T. suis isolates had identical sequences, and only 1 substitution was found in the ITS2 region of T. mobilensis. In contrast, the genus Trichomonas shared more diversity. The results obtained in this study support a possible future revision of the taxonomic classification of tritrichomonads.


Subject(s)
Genes, rRNA , Protozoan Infections, Animal , RNA, Ribosomal, 5.8S/genetics , Trichomonadida/classification , Trichomonadida/genetics , Trichomonas Infections/parasitology , Animals , Base Sequence , Cattle , Cattle Diseases/parasitology , DNA, Protozoan/analysis , DNA, Protozoan/genetics , DNA, Ribosomal/genetics , Humans , Molecular Sequence Data , Polymerase Chain Reaction/methods , Protozoan Infections/parasitology , Sequence Analysis, DNA , Swine , Swine Diseases/parasitology , Trichomonadida/isolation & purification , Trichomonas/genetics , Tritrichomonas/genetics
13.
Parasitology ; 115 ( Pt 2): 205-11, 1997 Aug.
Article in English | MEDLINE | ID: mdl-10190176

ABSTRACT

Myophilin is a muscle-associated antigen of the taeniid cestode Echinococcus granulosus. This protein shows a high amino acid sequence homology with calponins and calponin-like proteins, which are proposed to be associated with the regulation of smooth muscle contraction. In order to provide supportive evidence for a relationship between these proteins, we characterized myophilin using electrophoretic, biochemical and molecular biological approaches. Two-dimensional protein electrophoretic separation of E. granulosus larval proteins defined 4 isoelectric isoforms of myophilin (alpha, beta, gamma and delta), which appeared to be a consequence of post-translational modification of a single gene product. It was also demonstrated biochemically that E. granulosus myophilin undergoes specific phosphorylation in vitro by protein kinase C (PKC). Finally, myophilin homologues were identified in extracts of Taenia hydatigena and T. ovis by immunoblot. A partial cDNA of the closely related species, E. multilocularis, was isolated by cloning procedures and showed 99% homology with the E. granulosus myophilin gene. The similarities of E. granulosus myophilin with calponins in their tissue localization, protein isoforms patterns, ability to be phosphorylated in vitro by PKC, and the relatively conserved nature of the protein among related parasites suggest that myophilin may be associated with smooth muscle contraction.


Subject(s)
Antigens, Helminth , Echinococcus/chemistry , Helminth Proteins/metabolism , Muscle Proteins/metabolism , Protein Kinase C/metabolism , Animals , Blotting, Northern , DNA, Helminth , Echinococcosis/parasitology , Echinococcus/metabolism , Echinococcus/physiology , Electrophoresis, Gel, Two-Dimensional , Electrophoresis, Polyacrylamide Gel , Helminth Proteins/chemistry , Helminth Proteins/genetics , Immunoblotting , Muscle Proteins/chemistry , Muscle Proteins/genetics , Phosphorylation , Protein Isoforms , Recombinant Proteins/metabolism , Sequence Analysis, DNA
14.
Mol Biochem Parasitol ; 85(1): 113-24, 1997 Mar.
Article in English | MEDLINE | ID: mdl-9108553

ABSTRACT

Previous investigations had shown that the Giardia lamblia clone GS/M-83-H7-specific variant surface protein (VSP) H7 consists of at least two antigenically distinct parts: (i) a variable 314-aa N-terminal region which contains one, or more, variant-specific epitopes eliciting a transient and consequently low-level antibody response preferentially detectable during the early phase of G. lamblia infection in mice; and (ii) a 171-aa C-terminal region which contains relatively conserved epitope(s) causing a persistent and consequently high-level antibody response during the later phase of an infection. The present study indicated that monoclonal antibody G10/4 and polyclonal antibodies from early-phase infected or hyperimmunized mice, directed against the variant-specific N-terminal regional exclusively recognized conformational cysteine-containing epitopes. These antibodies caused detachment and aggregation of trophozoites, and exhibited complement-independent cytotoxic effect towards the parasite. In contrast, polyclonal antibodies from late-phase infected mice, directed against the semi-conserved peptidyl structures in the C-terminal region, preferentially reacted with non-conformational epitopes. Such antibodies had no cytotoxic effect, but provoked parasite-detachment and -aggregation. These findings indicated that infection of mice with G. lamblia clone GS/M-83-H7 generates a heterogeneous repertoire of cytologically active anti-VSP antibodies which may have a direct influence on the course of the parasite infection.


Subject(s)
Antibodies, Protozoan/immunology , Antigens, Protozoan/immunology , Antigens, Surface/immunology , Giardia lamblia/immunology , Protozoan Proteins , Animals , Antibodies, Protozoan/biosynthesis , Antigenic Variation , Binding Sites , Cytotoxicity, Immunologic , Enzyme-Linked Immunosorbent Assay , Epitopes , Female , Giardiasis/immunology , Immunoglobulin Isotypes , Mice , Recombinant Proteins/biosynthesis , Recombinant Proteins/immunology
15.
Parasitology ; 115 ( Pt 6): 581-90, 1997 Dec.
Article in English | MEDLINE | ID: mdl-9488869

ABSTRACT

Neospora caninum is an apicomplexan parasite of veterinary importance which invades many different cell types and tissues. N. caninum tachyzoites proliferate intracellularly by endodyogeny. Eventually the massive proliferation of tachyzoites leads to host cell lysis and the newly formed parasites are released and invade neighbouring cells. Tachyzoite cell surface molecules could serve as ligands, mediating host cell adhesion and invasion. Nc-p43 is a recently identified N. caninum tachyzoite surface protein which is functionally involved in the processes leading to host cell invasion in vitro. Affinity-purified antibodies directed against Nc-p43 were used to screen a lambda gt22A-cDNA expression library constructed from N. caninum tachyzoites. The cDNA insert of one immunoreactive clone was subcloned and expressed in E. coli as a poly-histidine fusion protein. The identity of the resulting recombinant antigen termed recNc-p43 was confirmed by immunoblotting, immunofluorescence and electron microscopy using affinity-purified antibodies. The sequence of the cDNA insert encoding recNc-p43 was determined. Analysis of the deduced amino acid sequence revealed that Nc-p43 exhibited similarity to SAG1 (p30) and SAG3 (p43), 2 major surface antigens of Toxoplasma gondii tachyzoites. These similarities were not reflected on the immunochemical level, since no cross-antigenicity between SAG1, SAG3 and Nc-p43 was observed.


Subject(s)
DNA, Protozoan/genetics , Neospora/genetics , Protozoan Proteins/genetics , Amino Acid Sequence , Animals , Antigens, Protozoan/chemistry , Antigens, Protozoan/genetics , Antigens, Surface/chemistry , Antigens, Surface/genetics , Chlorocebus aethiops , DNA Transposable Elements , DNA, Complementary , Fluorescent Antibody Technique , Gene Library , Host-Parasite Interactions , Immunoblotting , Immunohistochemistry , Membrane Glycoproteins/chemistry , Membrane Glycoproteins/genetics , Membrane Proteins/chemistry , Membrane Proteins/genetics , Membrane Proteins/immunology , Molecular Sequence Data , Neospora/chemistry , Neospora/immunology , Neospora/ultrastructure , Protozoan Proteins/chemistry , Protozoan Proteins/immunology , Toxoplasma/chemistry , Toxoplasma/genetics , Vero Cells
17.
Article in English | MEDLINE | ID: mdl-8833172

ABSTRACT

In this study, the conditions for the successful application of the random amplified polymorphic DNA (RAPD) assay to differentiate mite populations based on genetic variation were defined. Five species of mites related to allergic diseases were studied: Dermatophagoides pteronyssinus, D. farinae (2 strains), Blomia tropical is, Glycyphagus domesticus and Tyrophagus putrescentiae. The mites were isolated from pure cultures and processed according to the method described in this paper. The banding patterns obtained were different for all the species studied. When the DNA from two different strains of D. farinae were studied, the "fingerprint" banding patterns obtained showed differences between them. The random amplified polymorphic DNA assay may be a useful tool to aid the taxonomic study of mite populations.


Subject(s)
DNA/analysis , Hypersensitivity, Immediate/etiology , Mites/genetics , Animals , Genetic Variation/genetics , Mites/classification , Polymerase Chain Reaction , Random Amplified Polymorphic DNA Technique
18.
Schweiz Arch Tierheilkd ; 138(3): 139-43, 1996.
Article in German | MEDLINE | ID: mdl-8721188

ABSTRACT

"Arbitrarily primed PCR" (AP-PCR) methods are based on the amplification of DNA with arbitrarily selected primers. In the present review article two selected applications of this methodical approach are summarized. Using the RAPD ("Random Amplified Polymorphic DNA") technique the genetic variability of different organisms can be analyzed by generating a genomic fingerprint. Echinococcus granulosus isolates (metacestodes) from Spain and Switzerland were comparatively characterized by the use of this molecular epidemiological tool. The following groups of genetically related isolates could be identified: (a) equine isolates (horse/donkey) from Spain and Switzerland, (b) cattle isolates from Switzerland, (c) goat isolates from Spain and pig isolates from both countries, (d) sheep, cattle and human isolates from Spain. Isolates derived from Swiss patients were forming a separate group with respect to their genetic relatedness. As a further application of the AP-PCR, the development of a novel technique for the construction of cDNA libraries from minute amounts of starting material using an Echinococcus multilocularis metacestode library as an example is presented.


Subject(s)
Parasitic Diseases, Animal , Polymerase Chain Reaction/veterinary , Animals , Cattle , Cattle Diseases , Echinococcosis/diagnosis , Echinococcus/genetics , Echinococcus/isolation & purification , Equidae , Horse Diseases , Horses , Mice , Parasitic Diseases/diagnosis , Polymerase Chain Reaction/methods , Random Amplified Polymorphic DNA Technique/veterinary , Swine , Swine Diseases
19.
Schweiz Arch Tierheilkd ; 138(3): 144-51, 1996.
Article in German | MEDLINE | ID: mdl-8721189

ABSTRACT

The PCR is now widely introduced as a diagnostic and epidemiological tool in veterinary and human parasitology. Certain parasitic infections are detected using PCR with much higher sensitivity compared to conventional methods, and novel molecular approaches are considered for the analysis of infectiological questions. A broad spectrum of parasites relevant for veterinary and human parasitology already can be diagnosed by PCR. In the present review article, PCR-based methods developped or applied at the Institute of Parasitology in Berne for the detection of protozoan infections (Tritrichomonosis, Neosporosis, Toxoplasmosis) and helminthic infections (Echinococcosis, Taeniosis) are summarized.


Subject(s)
Helminthiasis, Animal , Polymerase Chain Reaction/veterinary , Protozoan Infections, Animal , Animals , Coccidiosis/diagnosis , Coccidiosis/veterinary , Echinococcosis/diagnosis , Echinococcosis/veterinary , Helminthiasis/diagnosis , Polymerase Chain Reaction/methods , Protozoan Infections/diagnosis , Taeniasis/diagnosis , Taeniasis/veterinary , Toxoplasmosis, Animal/diagnosis
20.
Parasitol Today ; 11(9): 320-6, 1995 Sep.
Article in English | MEDLINE | ID: mdl-15275313

ABSTRACT

Infection with the larval stage of the fox tapeworm Echinococcus multilocularis results in a life-threatening hepatic disease concerning humans and intermediate rodent hosts. Immunoepidemiological surveys provided information that a large proportion of infected individuals may demonstrate either constitutional resistance to early post-oncospheral development of the parasite or late resistance to disease by exhibiting an intrahepatic died-out parasite lesion. Similar events have been found in secondary infections of laboratory rodents. Dissection of humoral and cell-mediated immune responses in susceptible versus resistant individuals provides insight into immunological pathways associated with the different outcome of infection. Survival strategy of the metacestode obviously focuses on the crucial role played by the parasite laminated layer. This layer protects the metacestode from host effector mechanisms which can potentially kill the proliferating germinative compartments in case of resistant hosts. Bruno Gottstein and Richard Felleisen here discuss the need to search for more parameters discriminating between the different immune pathways in order to find out (immunogenetic?) predispositions responsible for the respective phenomena.

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