The status of intestinal parasitic infections and seroprevalence of cystic echinococcosis in the inhabitants in Dornod and Selenge, Mongolia, was observed with stool and serum samples from 165 and 683 residents from August 9 to August 15, 2003. A total of 10 inhabitants (6.1%) were found to be infected with protozoan cysts or oocysts by stool examinations; 7, 1 and 2 cases with Entamoeba coli, Giardia lamblia and Cryptosporidium parvum, respectively. A total of 62 people (9.1%) revealed significantly high levels of specific antibodies against Echinococcus granulosus by enzyme-linked immunosorbent assay (ELISA). The surveyed areas are rural areas and the sanitized tab water is not available. There is a possibility of endemic transmission of water-borne protozoan diseases in these areas. The clinical echinococcosis is needed to be searched from the seropositive inhabitants.
Protozoan Infections/*epidemiology/parasitology ; Mongolia/epidemiology ; Humans ; Giardia lamblia/isolation & purification ; Gastrointestinal Diseases/*epidemiology/*parasitology ; Entamoeba/isolation & purification ; Echinococcosis/*epidemiology ; Cryptosporidium parvum/isolation & purification ; Animals

Child ; Epstein-Barr Virus Infections ; complications ; Female ; Giardia lamblia ; isolation & purification ; Giardiasis ; complications ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Lymphohistiocytosis, Hemophagocytic ; drug therapy ; etiology

Giardia intestinalis infections arise primarily from contaminated food or water. Zoonotic transmission is possible, and at least 7 major assemblages including 2 assemblages recovered from humans have been identified. The determination of the genotype of G. intestinalis is useful not only for assessing the correlation of clinical symptoms and genotypes, but also for finding the infection route and its causative agent in epidemiological studies. In this study, methods to identify the genotypes more specifically than the known 2 genotypes recovered from humans have been developed using the intergenic spacer (IGS) region of rDNA. The IGS region contains varying sequences and is thus suitable for comparing isolates once they are classified as the same strain. Genomic DNA was extracted from cysts isolated from the feces of 5 Chinese, 2 Laotians and 2 Koreans infected with G. intestinalis and the trophozoites of WB, K1, and GS strains cultured in the laboratory, respectively. The rDNA containing the IGS region was amplified by PCR and cloned. The nucleotide sequence of the 3' end of IGS region was determined and examined by multiple alignment and phylogenetic analysis. Based on the nucleotide sequence of the IGS region, 13 G. intestinalis isolates were classified to assemblages A and B, and assemblage A was subdivided into A1 and A2. Then, the primers specific to each assemblage were designed, and PCR was performed using those primers. It detected as little as 10 pg of DNA, and the PCR amplified products with the specific length to each assemblage (A1, 176 bp; A2, 261 bp; B, 319 bp) were found. The PCR specific to 3 assemblages of G. intestinalis did not react with other bacteria or protozoans, and it did not react with G. intestinalis isolates obtained from dogs and rats. It was thus confirmed that by applying this PCR method amplifying the IGS region, the detection of G. intestinalis and its genotyping can be determined simultaneously.
Sequence Analysis, DNA ; Sensitivity and Specificity ; Polymerase Chain Reaction/*methods ; Phylogeny ; Mice ; Humans ; Giardiasis/parasitology/veterinary ; Giardia lamblia/*classification/genetics/*isolation & purification ; Genotype ; Dogs ; Dog Diseases/parasitology ; DNA, Ribosomal Spacer/*analysis ; DNA, Protozoan/*analysis/isolation & purification ; Base Sequence ; Animals

An epidemiological study on canine intestinal parasites was undertaken to evaluate changes in the prevalence among private household dogs from the Hachinohe region of Aomori prefecture, Japan, in 1997, 2002 and 2007, using the formalin-ethyl acetate sedimentation technique. The risk of zoonotic transmission from household dogs to humans was also discussed. All intestinal parasites detected in the present study (Giardia intestinalis, Isospora spp., Toxocara canis, Ancylostoma caninum, Trichuris vulpis and Strongyloides stercoralis) showed no changes in prevalence over the past 10 years based on analysis considering canine epidemiological profiles. In particular, prevalence of Giardia intestinalis in dogs under 1 year old, derived from pet shops/breeding kennels and kept indoors was unchanged, remaining at a high level of >15.0% at each time point. Toxocara canis also showed no changes in the group of dogs under 1 year old, bred by private owners and kept outdoors, and the prevalence was >10.0% every year. The present results indicate that the prevalence of Giardia intestinalis and other intestinal parasites in private household dogs has not always decreased, and the potential for direct parasitic zoonotic transmission from dogs to humans may be relatively high level, than from the environment (indoors and outdoors). We recommend careful surveillance of intestinal parasites and aggressive use of anthelminthic in private household dogs under considering the epidemiological factors.
Animal Husbandry ; Animals ; Anthelmintics/*therapeutic use ; Dog Diseases/drug therapy/epidemiology/*parasitology ; Dogs ; Giardia lamblia/*isolation & purification ; Humans ; Intestinal Diseases, Parasitic/drug therapy/epidemiology/parasitology/*veterinary ; Japan/epidemiology ; Parasitic Diseases, Animal/drug therapy/epidemiology/*parasitology ; Prevalence ; Time Factors ; Zoonoses

The cyclophilins (Cyps) are family members of proteins that exhibit peptidylprolyl cis-trans isomerase (PPIase, EC 5.2.1.8) activity and bind the immunosuppressive agent cyclosprin A (CsA) in varying degrees. During the process of random sequencing of a cDNA library made from Giardia intestinalis WB strain, the cyclophilin gene (gicyp 1) was isolated. An open reading frame of gicyp 1 gene was 576 nucleotides, which corresponded to a translation product of 176 amino acids (Gicyp 1). The identity with other Cyps was about 58-71%. The 13 residues that constituted the CsA binding site of human cyclophilin were also detected in the amino acid sequence of Gicyp 1, including tryptophan residue essential for the drug binding. The single copy of the gicyp 1 gene was detected in the G. intestinalis chromosome by southern hybridization analysis. Recombinant Gicyp 1 protein clearly accelerated the rate of cis-- Amino Acid Sequence ; Animals ; Cloning, Molecular ; Cyclophilins/antagonists & inhibitors/chemistry/genetics/*isolation&purification ; Cyclosporine/metabolism/pharmacology ; Giardia lamblia/*chemistry/genetics ; Human ; Immunosuppressive Agents ; Molecular Sequence Data ; Protein Binding ; Protozoan Proteins ; Recombinant Proteins

Non-human primates (NHPs) are confirmed as reservoirs of Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi. In this study, 197 fresh fecal samples from 8 NHP species in Qinling Mountains, northwestern China, were collected and examined using multilocus sequence typing (MLST) method. The results showed that 35 (17.8%) samples were positive for tested parasites, including Cryptosporidium spp. (3.0%), G. intestinalis (2.0%), and E. bieneusi (12.7%). Cryptosporidium spp. were detected in 6 fecal samples of Macaca mulatta, and were identified as C. parvum (n=1) and C. andersoni (n=5). Subtyping analysis showed Cryptosporidium spp. belonged to the C. andersoni MLST subtype (A4, A4, A4, and A1) and C. parvum 60 kDa glycoprotein (gp60) subtype IId A15G2R1. G. intestinalis assemblage E was detected in 3 M. mulatta and 1 Saimiri sciureus. Intra-variations were observed at the triose phosphate isomerase (tpi), beta giardin (bg), and glutamate dehydrogenase (gdh) loci, with 3, 1, and 2 new subtypes found in respective locus. E. bieneusi was found in Cercopithecus neglectus (25.0%), Papio hamadrayas (16.7%), M. mulatta (16.3%), S. sciureus (10%), and Rhinopithecus roxellana (9.5%), with 5 ribosomal internal transcribed spacer (ITS) genotypes: 2 known genotypes (D and BEB6) and 3 novel genotypes (MH, XH, and BSH). These findings indicated the presence of zoonotic potential of Cryptosporidium spp. and E. bieneusi in NHPs in Qinling Mountains. This is the first report of C. andersoni in NHPs. The present study provided basic information for control of cryptosporidiosis, giardiasis, and microsporidiosis in human and animals in this area.
Animals ; China ; Cryptosporidiosis/*parasitology ; Cryptosporidium/classification/genetics/*isolation & purification ; Enterocytozoon/classification/genetics/*isolation & purification ; Feces/parasitology ; Female ; Genotype ; Giardia lamblia/classification/genetics/*isolation & purification ; Giardiasis/parasitology/*veterinary ; Male ; Microsporidiosis/parasitology/*veterinary ; Molecular Sequence Data ; Phylogeny ; Primate Diseases/*parasitology ; Primates/classification/parasitology

Cryptosporidium and Giardia are 2 protozoan parasites responsible for waterborne diseases outbreaks worldwide. In order to assess the prevalence of these protozoans in drinking water samples in the northern part of Portugal and the risk of human infection, we have established a long term program aiming at pinpointing the sources of surface water, drinking water, and environmental contamination, working with the water-supply industry. Total 43 sources of drinking water samples were selected, and a total of 167 samples were analyzed using the Method 1623. Sensitivity assays regarding the genetic characterization by PCR and sequencing of the genes, 18S SSU rRNA, for Cryptosporidium spp. and beta,-giardin for G. duodenalis were set in the laboratory. According to the defined criteria, molecular analysis was performed over 4 samples. Environmental stages of the protozoa were detected in 25.7% (43 out of 167) of the water samples, 8.4% (14 out of 167) with cysts of Giardia, 10.2% (17 out of 167) with oocysts of Cryptosporidium and 7.2% (12 out of 167) for both species. The mean concentrations were 0.1-12.7 oocysts of Cryptosporidium spp. per 10 L and 0.1-108.3 cysts of Giardia duodenalis per 10 L. Our results suggest that the efficiency in drinking water plants must be ameliorated in their efficiency in reducing the levels of contamination. We suggest the implementation of systematic monitoring programs for both protozoa. To authors' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in drinking water samples in the northern part of Portugal.
Animals ; Cryptosporidium/*isolation & purification ; Cytoskeletal Proteins/genetics ; DNA, Protozoan/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Genes, rRNA ; Giardia lamblia/*isolation & purification ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Portugal ; Protozoan Proteins/genetics ; RNA, Protozoan/genetics ; RNA, Ribosomal, 18S/genetics ; Risk Assessment ; Sequence Analysis, DNA ; Water/*parasitology

The objective of this study was to genetically characterize isolates of Giardia duodenalis and to determine if zoonotic potential of G. duodenalis could be found in stray cats from urban and suburban environments in Guangzhou, China. Among 102 fresh fecal samples of stray cats, 30 samples were collected in Baiyun district (urban) and 72 in Conghua district (suburban). G. duodenalis specimens were examined using light microscopy, then the positive specimens were subjected to PCR amplification and subsequent sequencing at 4 loci such as glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi), beta-giardin (bg), and small subunit ribosomal RNA (18S rRNA) genes. The phylogenetic trees were constructed using obtained sequences by MEGA5.2 software. Results show that 9.8% (10/102) feline fecal samples were found to be positive by microscopy, 10% (3/30) in Baiyun district and 9.7% (7/72) in Conghua district. Among the 10 positive samples, 9 were single infection (8 isolates, assemblage A; 1 isolate, assemblage F) and 1 sample was mixed infection with assemblages A and C. Based on tpi, gdh, and bg genes, all sequences of assemblage A showed complete homology with AI except for 1 isolate (CHC83). These findings not only confirmed the occurrence of G. duodenalis in stray cats, but also showed that zoonotic assemblage A was found for the first time in stray cats living in urban and suburban environments in China.
Animals ; Cat Diseases/*parasitology ; Cats ; China ; Cluster Analysis ; DNA, Protozoan/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Feces/parasitology ; Giardia lamblia/*classification/cytology/genetics/*isolation & purification ; Giardiasis/parasitology/*veterinary ; Microscopy ; Molecular Sequence Data ; Phylogeny ; Protozoan Proteins/genetics ; RNA, Ribosomal, 18S/genetics ; Sequence Analysis, DNA

The objective of this study was to genetically characterize isolates of Giardia duodenalis and to determine if zoonotic potential of G. duodenalis could be found in stray cats from urban and suburban environments in Guangzhou, China. Among 102 fresh fecal samples of stray cats, 30 samples were collected in Baiyun district (urban) and 72 in Conghua district (suburban). G. duodenalis specimens were examined using light microscopy, then the positive specimens were subjected to PCR amplification and subsequent sequencing at 4 loci such as glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi), beta-giardin (bg), and small subunit ribosomal RNA (18S rRNA) genes. The phylogenetic trees were constructed using obtained sequences by MEGA5.2 software. Results show that 9.8% (10/102) feline fecal samples were found to be positive by microscopy, 10% (3/30) in Baiyun district and 9.7% (7/72) in Conghua district. Among the 10 positive samples, 9 were single infection (8 isolates, assemblage A; 1 isolate, assemblage F) and 1 sample was mixed infection with assemblages A and C. Based on tpi, gdh, and bg genes, all sequences of assemblage A showed complete homology with AI except for 1 isolate (CHC83). These findings not only confirmed the occurrence of G. duodenalis in stray cats, but also showed that zoonotic assemblage A was found for the first time in stray cats living in urban and suburban environments in China.
Animals ; Cat Diseases/*parasitology ; Cats ; China ; Cluster Analysis ; DNA, Protozoan/chemistry/genetics ; DNA, Ribosomal/chemistry/genetics ; Feces/parasitology ; Giardia lamblia/*classification/cytology/genetics/*isolation & purification ; Giardiasis/parasitology/*veterinary ; Microscopy ; Molecular Sequence Data ; Phylogeny ; Protozoan Proteins/genetics ; RNA, Ribosomal, 18S/genetics ; Sequence Analysis, DNA

This study aimed to identify the assemblages (or subassemblages) of Giardia duodenalis by using normal or nested PCR based on 4 genetic loci: glutamate dehydrogenase (gdh), triose phosphate isomerase (tpi), beta-giardin (bg), and small subunit ribosomal DNA (18S rRNA) genes. For this work, a total of 216 dogs' fecal samples were collected in Guangdong, China. The phylogenetic trees were constructed with MEGA5.2 by using the neighbor-joining method. Results showed that 9.7% (21/216) samples were found to be positive; moreover, 10 samples were single infection (7 isolates assemblage A, 2 isolates assemblage C, and 1 isolate assemblage D) and 11 samples were mixed infections where assemblage A was predominant, which was potentially zoonotic. These findings showed that most of the dogs in Guangdong were infected or mixed-infected with assemblage A, and multi-locus sequence typing could be the best selection for the genotype analysis of dog-derived Giardia isolates.
Animals ; China ; Cluster Analysis ; Coinfection/parasitology/veterinary ; Cytoskeletal Proteins/genetics ; DNA, Protozoan/chemistry/genetics ; Dog Diseases/parasitology ; Dogs ; Genotype ; Giardia lamblia/*classification/*genetics/isolation & purification ; Giardiasis/parasitology/*veterinary ; Glutamate Dehydrogenase/genetics ; Molecular Sequence Data ; *Multilocus Sequence Typing ; Phylogeny ; Polymerase Chain Reaction ; RNA, Ribosomal, 18S/genetics ; Triose-Phosphate Isomerase/genetics