Multiple-subgenotype infections of Giardia intestinalis detected in Palestinian clinical cases using a subcloning approach.

To evaluate the geographic distribution of Giardia intestinalis genotypes in Nablus, West Bank, Palestine, a genotyping study was performed using clinical fecal samples. Microscopic examination confirmed that 8 of 69 (11.6%) samples were G.

Molecular evidence of Enterocytozoon bieneusi in Japan.

Enterocytozoon bieneusi is an emerging and clinically significant enteric pathogen in humans associated mainly with chronic diarrhea. It has been found in a variety of wild, domestic and companion mammals and birds. To date, epidemiological surveys of E.

Genetical survey of novel type of Cryptosporidium andersoni in cattle in Japan.

Previously, we reported that an isolate of novel type of Cryptosporidium andersoni detected in cattle in Japan contained Type A (identical to C. andersoni reported previously) and Type B (having a thymine nucleotide insertion unlike the Type A) genotypes in the 18S rRNA gene. Here, we conducted an extensive investigation of Cryptosporidium infections in adult cattle in Japan from 2004 to 2007.

First record of Eimeria furonis infection in a ferret, Japan, with notes on the usefulness of partial small subunit ribosomal RNA gene sequencing analysis for discriminating among Eimeria species.

Eimeria is a genus of apicomplexan parasites found in a variety of vertebrates including the weasel. At present, three species have been reported in members of the weasel family, but the presence of weasel Eimeria in Japan have been quite unclear. The identification of Eimeria species has been performed based on oocyst morphology, host species, and habitat in the host, but sometimes discriminating among morphologically similar species under light microscopy is impossible.

Detection of a mixed infection of a novel Cryptosporidium andersoni and its subgenotype in Japanese cattle.

Cryptosporidium oocysts were detected in the feces of cattle in Saga, Japan. Isolates were morphologically large. We attempted to identify the species or genotypes of the isolates by analyzing the partial sequences of the 18S rRNA and Cryptosporidium oocyst wall protein (COWP) genes, and measuring the infectivity in mice.

Subgenotype analysis of Cryptosporidium parvum isolates from humans and animals in Japan using the 60-kDa glycoprotein gene sequences.

Cryptosporidium parvum is a well-known intestinal parasite which is associated with severe acute diarrhea in humans and animals. This parasite is composed of morphologically identical but genetically different multiple genotypes. In humans, cryptosporidiosis is mainly caused by two C.

Gene analysis of Cryptosporidium parvum HNJ-1 strain isolated in Japan.

We analyzed genetically Cryptosporidium parvum HNJ-1 strain, which is the Japanese reference strain isolated from human in Japan. DNA sequences of genes for thrombospondin-related adhesive protein of Cryptosporidium-1 and Cryptosporidium-2 (TRAP-C1, TRAP-C2), heat shock protein 70 (HSP70), oocyst wall protein (COWP), beta-tubulin, alpha-tubulin, polythreonine-region (Poly-T), elongation factor 1 alpha (EF-1 alpha), and 18S rRNA of this strain were determined. They showed high rate of homology to published sequences of genotype 2 strains, which were considered to be infective to both humans and animals.

Zoonotic genotype of Giardia intestinalis detected in a ferret.

Giardia intestinalis has been found in a variety of mammals, including humans, and consists of host-specific and zoonotic genotypes. There has been only 1 study of G. intestinalis infection in weasels, but the genotype of its isolate remains unclear.

Infectivity of a novel type of Cryptosporidium andersoni to laboratory mice.

Previously, we reported 'a novel type' of Cryptosporidium andersoni detected from cattle in Japan, and showed that the isolate was infective to mice. In the present study, we examined the patterns of oocyst shedding in both immunocompromised and immunocompetent mice, as well as pathological lesions in the infected mice. After oral inoculation with 1 x 10(6) oocysts, all five severe combined immunodeficiency (SCID) mice began to shed endogenously produced oocysts on day 6 post-inoculation (p.

Cross-reactivities with Cryptosporidium spp. by chicken monoclonal antibodies that recognize avian Eimeria spp.

In a previous study, we have developed several chicken monoclonal antibodies (mAbs) against Eimeria acervulina (EA) in order to identify potential ligand molecules of Eimeria. One of these mAbs, 6D-12-G10, was found to recognize a conoid antigen of EA sporozoites and significantly inhibited the sporozoite invasions of host T lymphocytes in vitro. Furthermore, some of these chicken mAbs showed cross-reactivities with several different avian Eimeria spp.

First record of Cryptosporidium infection in a raccoon dog (Nyctereutes procyonoides viverrinus).

Cryptosporidium species have been found in more than 150 species of mammals, but there has been no report in raccoon dogs. Here we found the Cryptosporidium organism in a raccoon dog, Nyctereutes procyonoides viverrinus, and identified this isolate using PCR-based diagnostic methods. Cryptosporidium diagnostic fragments of the 18S ribosomal RNA, Cryptosporidium oocyst wall protein and 70-kDa heat shock protein genes were amplified from the isolate and sequenced to reveal the phylogenetic relationships between it and other Cryptosporidium species or genotypes reported previously.

Molecular characterization of a Cryptosporidium isolate from a banded mongoose Mungos mungo.

Cryptosporidium spp. has been found in more than 150 species of mammals, but there has been no report in mongooses. In this study, we report the isolation of Cryptosporidium sp.

Identification of Cryptosporidium isolates from cockatiels by direct sequencing of the PCR-amplified small subunit ribosomal RNA gene.

Cryptosporidium is a significant pathogen in humans and animals. Cases of infection by C. meleagridis or C.

Polymerase chain reaction-based genotype classification among human Blastocystis hominis populations isolated from different countries.

Since the genotype of human Blastocystis hominis isolates is highly polymorphic, PCR-based genotype classification using known sequenced-tagged site (STS) primers would allow the identification or classification of different genotypes. Five populations of human B. hominis isolates obtained from Japan, Pakistan, Bangladesh, Germany, and Thailand were subjected to genotype analysis by using seven kinds of STS primers.

Recombinant bovine herpesvirus-1 expressing p23 protein of Cryptosporidium parvum induces neutralizing antibodies in rabbits.

In order to develop a vaccine against cryptosporidiosis in cattle, we constructed a recombinant bovine herpesvirus-1 (BHV-1) expressing an immunodominant surface protein, p23, of Cryptosporidium parvum sporozoites. In the recombinant virus, the p23 gene under the control of a CAG promoter and a gene coding for an enhanced green fluorescent protein were integrated into the gG gene of BHV-1. Despite a low frequency of homologous recombination, cloning of the recombinants was easy because of the specific fluorescence of the plaques formed by recombinants.

[Identification of species of Cyclospora isolates from patients by the PCR-based diagnostic methods].

Cyclospora cayetanensis is an intestinal coccidian parasite and known as a human pathogen causing watery diarrhea. Recently, Cyclospora organisms, morphologically indistinguishable from C. cayetanensis, were detected from the several species of primates, and three new species named, C.

Identification of genotypes of Cryptosporidium parvum isolates from ferrets in Japan.

Recent molecular studies show that Cryptosporidium parvum is composed of at least eight genotypes including zoonotic genotypes. Therefore, it is important to screen the genotypes of the isolates harbored in animals for the control of human cryptosporidiosis. The ferret is a popular pet, but also a reservoir of Cryptosporidium.

Identification of genotypes of Giardia intestinalis isolates from dogs in Japan by direct sequencing of the PCR amplified glutamate dehydrogenase gene.

Giardia has been detected in domestic dogs in Japan, but the genotype of isolates has remained unclear because identification has relied on conventional microscopy. Here we tried to identify the genotypes of four isolates from dogs in Japan by direct sequencing of the PCR amplified Giardia glutamate dehydrogenase (GDH) gene. The primer pair GDHF3 and GDHB5, targeting the GDH gene, was designed to prime a region of the GDH gene sequence conserved in the strains found to have the dog-specific genotype.

[Usefulness of multiplex-PCR for identification of Entamoeba histolytica and Entamoeba dispar].

We evaluated the usefulness of a multiplex-PCR method for differentiation of Entamoeba histolytica and Entamoeba dispar, which are morphologically indistinguishable species. Cultured trophozoites of E. histolytica HM-1: IMSS and E.

[Comparative study of PCR-based Cryptosporidium discriminating techniques with a review of the literature].

We identified the species or genotypes of the six Cryptosporidium isolates from patients and C. parvum strain HNJ-1 using the seven previously described species-differentiation and genotyping PCR protocols for detection of Cryptosporidium parasites. In addition, we also discussed about the usefulness of these PCR-based protocols on the basis of the reports previously published.

Cryptosporidium infection in dogs in Osaka, Japan.

Cryptosporidium parvum is a zoonotic pathogen composed of genetically distinct but morphologically identical genotypes. Recent molecular study indicates that dogs may transmit the cattle genotype, which is known to be pathogenic to humans. Although large-scale studies of Cryptosporidium infection in dogs have been performed in several countries, the isolates were not accurately identified because of the lack of a method for molecular analysis.