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1.
Our previous report demonstrated that small wild rodents in Japan harbored two types of novel Babesia microti-like parasites (Kobe and Hobetsu types), but not the type widely distributed throughout the temperate zones of North American and Eurasian Continents (U.S. type). In this study, we surveyed small wild mammals collected at various places in the northern part of Japan, seeking for U.S.-type B. microti. A total of 197 small mammals comprising 10 species, Apodemus speciosus, A. argenteus, Clethrionomys rufocanus, C. rutilus, Eothenomys andersoni, Microtus montebelli, Tamias sibiricus, Sorex unguiculatus, S. caecutiens, and Urotrichus talpoides, were examined. Babesia parasites were detected in A. speciosus, C. rufocanus, C. rutilus, M. montebelli, S. unguiculatus, and S. caecutiens by microscopy of blood smears and by PCR targeting babesial nuclear small-subunit rRNA (rDNA) and beta-tubulin genes. Inoculation of their bloods into experimental animals gave rise to 23 parasite isolates, which included 16 from A. speciosus, 4 from C. rufocanus, and 1 each from C. rutilus, M. montebelli and S. unguiculatus. Sequencing analyses of their rDNA and beta-tubulin genes revealed that, of the 23 isolates, 20 and 3 were of Hobetsu and U.S. types, respectively. The U.S.-type B. microti strains isolated in Japan, however, were distinguishable from the isolates in the United States when their beta-tubulin gene sequences and antigen profiles in Western blots were compared. We conclude that U.S.-type B. microti exists in Japan although it has been genetically and antigenically diversified from that distributed in the United States. The results also suggest that not only rodents, but also some insectivores may serve as reservoirs for the agent of human babesiosis.  相似文献   

2.
A total of 1,335 archived human sera collected in 1985 from an area in Japan where a tick-borne disease is endemic were examined by indirect immunofluorescence antibody test (IFAT) to estimate seroprevalence against three serologically distinct types of Babesia microti-like parasites; namely, Hobetsu, Kobe, and U.S. types. Eighteen sera (1.3%) were found to be IFAT-positive (titer 1:100-1:6,400), of which 14 and three were ascertained by Western blot analysis to be positive against the Hobetsu and Kobe types, respectively. In addition, four sera showed an IFAT titer of 1:100 against the U.S. type, but they appeared to be false-positive because they were cross-reactive against the Hobetsu and Kobe types, and also because a U.S.-type parasite has not been found in Japan. Our results suggest that human babesiosis in Japan occurred prior to the discovery of the index case in 1999 and that the infections were caused mainly by Hobetsu-type parasites.  相似文献   

3.
The first case of human babesiosis was reported in Japan. The epidemiology of this disease in Japanese nature remains unclear. In this study, 97 common field mice captured in Hokkaido, Japan, were examined. Blood specimens absorbed onto filter papers were eluted and tested by nested PCR using specific primers for the B. microti nuclear small subunit rRNA genome. Twenty-three percent (11/47) of Apodemus speciosus and four percent (2/50) of Clethrionomys rufocanus were positive. The 159-bp primary sequences of PCR products tested exhibited 97.5% and 96.8% homology with those of the human isolate in Japan and of U.S. strains of B. microti, respectively.  相似文献   

4.
Nucleotide sequences of ribosomal DNA (rDNA) of Babesia (B.) gibsoni occurring in Miyazaki, western Japan, were examined using blood samples obtained from seven dogs suffering from natural canine babesiosis. DNA isolated from these blood samples was subjected to the polymerase chain reaction (PCR). The nucleotide sequences of the PCR products were determined and compared with other rDNA sequences of B. gibsoni isolated from Asia, Europe and U.S.A. Although homology values between our isolates and those isolated from Europe and U.S.A. were both 84.0%, respectively, our isolates were identical to the Asian types. In conclusion, B. gibsoni occurring in Miyazaki was revealed to have the genotype Asia 1 or Asia 2 from a comparison of the partial rDNA sequences.  相似文献   

5.
Six Eurasian red squirrels (Sciurus vulgaris orientis), victims of road traffic found during 2002 and 2004 near the Noppro Forest Park in Ebetsu, Hokkaido, Japan, were examined for the presence of Babesia parasites. Three of the six squirrels exhibited positive signals by nested PCRs targeting both the 18S rRNA and beta-tubulin genes. Three squirrels proved to be infected with a B. microti-like parasite as evidenced by sequencing the amplified DNAs and by the morphology of the intraerythrocytic parasites. Genotypically, however, the parasite appeared to be of a new type, as it was clearly distinguishable from any of the known types that have previously been reported in various wild animals. This is the first report showing molecular evidence for the presence of B. microti-like parasites in Sciuridae.  相似文献   

6.
A 1344 bp fragment of the 16S ribosomal DNA (rDNA) sequence was used to determine the genetic relationship of Pasteurella pneumotropica isolates from laboratory rodents. A total of 30 nucleotide sequences of P. pneumotropica, including 24 wild strains, 3 reference strains, and 3 nucleotide sequences deposited in GenBank, were examined for heterogeneity of their 16S rDNA sequences. Phylogenetic analysis based on 16S rDNA sequence discriminated 5 types of branching lineages. Of these 5 types, 3 types had significant associations with mice or rats, and 2 had significant associations with the beta-hemolytic phenotype. These results suggest that 16S rDNA sequencing of P. pneumotropica isolates demonstrates genetic heterogeneity and phylogenetic discrimination in terms of their hemolytic phenotype and host associations.  相似文献   

7.
To differentiate the morphologically similar pinworms of the common laboratory rodents, such as Syphacia obvelata and Syphacia muris, we amplified and sequenced the region spanning the internal transcribed spacer 1 (ITS-1), 5.8S gene, and ITS-2 of the ribosomal DNA followed by designing of species-specific primers for future use in the identification of the worms. It was observed that S. obvelata, S. muris and Aspiculuris tetraptera can be differentiated from each other based on their rDNA sequences. This is the first report of the ITS-1, 5.8S, and ITS-2 of the rDNA of the three aforementioned rodent pinworm species. The use of restriction endonucleases, AluI or RsaI, further allowed the delineation of the three species. Moreover, we also constructed species-specific primers that were designed for unique regions of the ITS-2 of the three species. This approach allowed their specific identification with no amplicons being amplified from heterogenous DNA samples, and sequencing confirmed the identity of the sequences amplified. Thus, the use of these specific primers along with PCR-RFLP can serve as useful tools for the identification of pinworms in rats, mice, and wild rodents.  相似文献   

8.
Small babesiae in dogs are generally considered to belong to Babesia gibsoni. Here we describe the genotypic characterisation of small piroplasms found in the blood of a dog which suffered from clinical babesiosis. Pairwise identities as well as distance, parsimony and maximum likelihood analyses of the 18S rDNA clearly demonstrated that this isolate was only distantly related to the other canine piroplasms characterised genetically so far, including B. gibsoni. It was more closely related to B. microti, B. rodhaini, and Theileria equi. It is concluded that the small canine piroplasms described in this study represent a hitherto unknown species and that the fauna of piroplasms occurring in dogs is more diverse than assumed so far.  相似文献   

9.
Sun Y  Liu G  Yang L  Xu R  Cao W 《Veterinary parasitology》2008,156(3-4):333-339
A Babesia microti-like rodent parasite was isolated from the tick, Ixodes persulcatus, collected from the northern forest area of Heilongjiang province, China. The collected I. persulcatus were allowed to feed on specific pathogen-free SCID mice and red blood cells from the mice were used to isolate Babesia spp. with the microareophilous stationary-phase culture technique. Paired and tetrad forms of merozoites were observed by light microscope in red blood cells of SCID mice. In vitro growth of the parasites was also achieved in mice erythrocytes, which indicated the presence of Babesia spp. in I. persulactus. To further identify the Babesia species, polymerase chain reaction screening and subsequent sequencing of nuclear small subunit ribosomal RNA (nss-rRNA) was employed. The results indicate that the observed parasites might be an isolate strain responsible for human babesiosis -B. microti - which has 99.3% identity with that of B. microti isolate RcM5201 (AB112050) from Mishan in Heilongjiang and Kobe isolates from Japan. In addition, the infection rate of B. microti in I. persulcatus ticks in the region was 3.6-4.0% in adult females and no infection in males. Though the infection rate is low, the high attack frequency of tick species on local residents indicates the risk of human babesiosis in the region and the necessity of precautionary measures.  相似文献   

10.
Piroplasms are tick-transmitted blood parasites belonging to the genera Babesia and Theileria. In western and southern Switzerland, B. divergens, a small Babesia species, has been known for a long time as a parasite of cattle. Recent investigations have revealed the autochthonous occurrence of this parasite also in central and eastern Switzerland. On the occasion of an outbreak of anaplasmosis in the canton of Grisons, however, B. bigemina, a large Babesia species, and Theileria of the buffeli/sergenti/orientalis species complex were for the first time identified; the epidemiology of these two piroplasms in Switzerland remains unknown until now. The recent identification by genetic analyses of B. divergens in wild ruminants contradicts the hitherto postulated strict host specificity of this Babesia species for cattle. B. divergens as well as the closely related Babesia spp. genotype EU1 have in single cases also been identified in splenectomized humans.The rodent babesia B. microti which causes a human infection that is considered an "emerging tick-borne disease" in the U.S.A., is widespread in rodent populations in Switzerland, but seems to be of minor relevance as zoonotic pathogen here. Reasons for this could be differences in virulence of the parasites or in the transmission by the respective tick-vectors on the two continents.  相似文献   

11.
The heat shock protein 70 (hsp70) genes of Babesia gibsoni, B. canis canis, B. canis vogeli, and B. canis rossi isolated from infected dogs were cloned by polymerase chain reaction (PCR) and sequenced. In the nucleotide sequence and the predicted amino acid sequence of the gene, the parasites were very similar to each other. The nucleotide sequences of the hsp70 gene had more variety than those of 18S nuclear subunit ribosomal DNA (18S rDNA). A phylogenetic analysis of these sequences and comparisons with sequences from other Babesia and Theileria species revealed that all canine babesial isolates analyzed in the present study were closely related to each other and formed one cluster. Additionally, a phylogenetic analysis of Babesia and Theileria species showed that these parasites could be divided into three groups: group A including canine babesial isolates, B. divergens, B. odocoilei, B. bovis, B. caballi, and B. ovis; group B including Theileria annulata, T. orientalis, and T. cervi; and group C including B. microti and B. rodhaini. These results suggested that a phylogenetic analysis of the hsp70 gene sequence might be helpful in classifying Babesia and Theileria species, and that canine babesial isolates might be closely related to each other, indicating their evolution from the same ancestry.  相似文献   

12.
Changes of splenic lymphocyte subpopulation after Babesia microti and Babesia rodhaini inoculation in mice were examined by flow cytometric analysis. The B. microti inoculated mice showed a longer period of time from inoculation to the onset of increase or decrease parasitaemia (%), packed cell volume, total spleen cell numbers and surface immunoglobulin positive splenic cell numbers than respective periods in B. rodhaini inoculated mice. The Thy-1 positive cell numbers in B. microti inoculated mice and B. rodhaini inoculated mice pre-immunized with homologous parasites were significantly higher than that of B. rodhaini inoculated mice. The ratio of L3T4 positive cell/Lyt-2 positive cell after inoculation with B. microti was quite similar to that in B. rodhaini mice pre-immunized. However, the ratio in B. rodhaini inoculated mice revealed a lack of an increasing phase. These results suggested that the T-cell dependent early immune response, especially suppressor activity, was closely related to the difference in the course of infection between the non-lethal B. microti and the lethal B. rodhaini infection in mice.  相似文献   

13.
The hosts of larvae and nymphs of Amblyomma tigrinum, a tick whose adults feed on wild and domestic Canidae in South America, are uncertain. A 17 months survey was carried out trapping wild vertebrates in north-western Córdoba, Argentina, to evaluate their parasitism with A. tigrinum subadults. Larvae and nymphs of this tick species were identified conventionally and by comparison of 16S rDNA sequences with GenBank deposited sequences. A total of 207 small and medium-sized rodents and 182 birds were captured and examined for ticks. Most ticks on birds were from ground forest feeding birds (BB) with a minimal contribution of birds feeding in open pastures. All ticks from rodents were obtained from representatives of the families Cricetidae (SR) and Caviidae (MR). Percent of larvae infestation was higher (P<0.01, Chi-square distribution) in BB (55.2%) and SR (46.4%) than in MR (17.4%) and the same trend was found for number of larvae on these hosts (test of Kruskal-Wallis). Caviidae (only representative Galea musteloides) rodents were extremely prone to be infested with nymphs of A. tigrinum (94.2%) followed by BB (50.6%) and SR (3.6%) (P<0.01) and the same tendency was found for number of nymphs (P<0.01). The index of aggregation for nymphs on MR was the lowest (0.409) followed by nymphs on BB (0.706) which may be a consequence of higher and homogenous exposure of G. musteloides to host-seeking nymphs. Several BB are food source for both larvae and nymphs of A. tigrinum while for rodents larvae were common only on SR (mainly on the Sigmodontinae Akodon dolores and Graomys sp.) and nymphs feed almost exclusively on MR. Therefore, both birds and rodents are of importance for the survival strategy of A. tigrinum subadults. The plasticity of A. tigrinum to colonize areas with different climates plus the capacity of their subadults to feed on hosts widely distributed indicates that this tick has the potential to become a widespread parasite but this does not seem to be the actual situation. Several proposals are presented to further understand its ecology.  相似文献   

14.
During the 1984 to 1986 spring hunting seasons in Connecticut, viscera from 300 hunter-killed wild turkeys and blood samples from live-trapped wild turkeys were examined in order to establish a health profile on the State's wild turkey population. Seven species of endoparasites were recovered from 224 (75%) of 300 birds: Metroliasthes lucida, Ascaridia dissimilis, Heterakis gallinarum, Syngamus trachea, Capillaria species, Trichomonas gallinarum, and Eimeria species. The most prevalent parasites were A. dissimilis (52%) and M. lucida (37%). Although some turkeys harbored high intensities of these two helminths, there were no associated gross or microscopic lesions nor body weight changes. The prevalence of S. trachea, H. gallinarum, Capillaria and Eimeria species, which are potential pathogenic parasites in domestic and wild turkeys, was very low (less than 3%). Blood samples from 19 live-trapped wild turkeys were negative for hemoprotozoa and antibodies to 15 common bacterial and viral agents. Serum samples from 82 birds were negative for Mycoplasma gallisepticum and Mycoplasma synoviae. The survey indicates that the wild turkey population of Connecticut presently has little evidence of common infectious diseases and minimal prevalence of potentially pathogenic parasites.  相似文献   

15.
Serologic and molecular evidence suggest that white-tailed deer in South Texas and North Mexico carry the agents of bovine babesiosis, Babesia bovis and Babesia bigemina. To determine if white-tailed deer in central Texas, which is outside the known occurrence of the vector tick at this time, harbor these parasites, blood samples from free-ranging and captive white-tailed deer (Odocoileus virginianus) in Tom Green County were tested by polymerase chain reaction (PCR) assays for B. bovis and B. bigemina 18S rDNA. Of the 25 samples tested, three (12%) were positive by nested PCR for B. bovis. This identity was confirmed by sequence analysis of the cloned 18S rDNA PCR product. Further confirmation was made by sequence analysis of the rRNA internal transcribed spacer (ITS) 1, 5.8S rRNA gene, and ITS 2 genomic region in two (representing samples from two different ranches) of the B. bovis positive samples. Three samples were positive by B. bigemina nested PCR, but sequencing of the cloned products confirmed only one animal positive for B. bigemina; Theileria spp. DNA was amplified from the other two animal samples. In addition to Theileria spp., two genotypically unique Babesia species sequences were identified among the cloned sequences produced by the B. bigemina primers in one sample. Phylogenetic analysis showed no separation of the deer B. bovis or B. bigemina 18S rDNA, or deer B. bovis ITS region sequences from those of bovine origin. Clarification of the possible role of white-tailed deer as reservoir hosts in maintaining these important pathogens of cattle is critical to understanding whether or not deer contribute to the epidemiology of bovine babesiosis.  相似文献   

16.
Blood cell panasitaemias of Babesai microti and associated haematological changes were studied in mice concurrently infected with Schistosoma mansoni and in controls with no schistosome infections. In comparison to the controls B. microti parasit-aemias were suppressed markedly in mice harbouring 6 and 8 weeks old patent S. mansoni infections at the time of infection with B. microti. The suppression was accompanied by an alleviation/annulment of the B. microti induced reductions in haemoglobin and haematocrit levels and in its erythropoetic stimulation. The suppressive effect of patent S. mansoni infections on B. microti is suggested to be at least partly attributable to non-specific immunological factors although the altered erythrocyte kinetic state induced by the schistosoma infection in combination with the preference of B. microti for mature (old) erythrocytes might also play a role in the suppression. In mice harbouring 2 weeks old prepatent S. mansoni infections B. microti was not suppressed.  相似文献   

17.
This study investigated the differences in Salmonella isolation rates in environmental samples taken from several types of hen houses in Chiba, Japan. In addition, for the detailed epidemiologic survey, environmental samples, hens, and rodents were collected from Salmonella-contaminated windowless houses on three farms. As a result, Salmonella was isolated from four (80%) of five farms with windowless hen houses; Salmonella enteritidis was isolated from a single windowless house. In contrast, only one serotype of Salmonella was isolated from 1 (6.7%) of 15 farms with open hen houses. In the S. enteritidis-contaminated windowless hen house, the isolation rates of S. enteritidis as compared with the other serotypes were 90.9% of environments, 94.1% of hens, and 86.4% of roof rats (Rattus rattus) that resided in the environments. In reference to the phage type (PT) of these isolates, PT1 was detected in environments and roof rats, and PT9 was detected in both these samples and in hens. Thus, the Salmonella isolation rate in hen houses seems to be associated with whether the premises are windowless or open. Moreover, roof rats appear to be the most important vectors in the spread of S. enteritidis in the windowless hen house because the S. enteritidis PTs coincide with each other.  相似文献   

18.
An epidemiological survey on a Theileria parasite infection of cattle in Northeast China was carried out using allele-specific PCR and DNA sequence analysis of the major piroplasm surface protein (MPSP) gene. The results showed that 14 of 104 blood samples were positive for Theileria by PCR. Among the positive cases, co-infection with various combinations of C- and I-type parasites was detected in 12 samples; no B- and Thai-type parasites were detected by allele-specific PCR. Phylogenetic analysis based on the MPSP gene sequences revealed that Theileria parasites with the MPSP types 1, 2, and 4 were distributed in Northeast China.  相似文献   

19.
Microscopic examination of Giemsa-stained peripheral blood smears collected from three naturally infected dogs originating from Turkey revealed the presence of large (around 4.5-5.0 microm) intraerythrocytic Babesia parasites in all dogs. DNA was extracted from the three infected blood samples and an around 410 bp portion of the 18S rDNA gene of Babesia species was PCR amplified for subsequent molecular characterization. RFLP analysis of the PCR products suggested the presence of the species B. vogeli in all infected dogs and sequencing of the PCR products from two of the three samples revealed 100% identity among the two Turkish isolates. Comparisons with the equivalent 410 bp portions of the 18S rDNA gene of Babesia species confirmed the affiliation of these isolates to the B. vogeli species. This is the first report and molecular characterization of dog infection with a large Babesia species in Turkey.  相似文献   

20.
Raccoons (Procyon lotor), which have recently become feral in Japan, were examined for the presence of Babesia microti-like parasites. Out of 372 raccoons captured in the west-central part of Hokkaido, 24 animals with splenomegaly were selected and tested by nested PCR targeting the babesial 18S rRNA gene. B. microti-like parasites were detected in two of the 24 individuals, and their DNA sequences were identical to that of the B. microti-like parasite reported from raccoons in the United States, suggesting that the parasites were probably imported into Japan and that the life cycle of the parasite has already been established in the country. The potential risk of this B. microti-like parasite spreading among dogs and foxes in Japan will need to be carefully monitored, as parasitization by phylogenetically very close parasites has been reported from such animals.  相似文献   

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