Detection of Babesia microti-like parasite in filter paper-absorbed blood of wild rodents.
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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. (+info)
Transmission studies of Babesia microti in Ixodes ricinus ticks and gerbils.
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In order to investigate the possible role of Ixodes ricinus as a vector of zoonotic Babesia microti infection in Europe, a European rodent isolate (HK) and a zoonotic American isolate (GI) were studied in transmission experiments. PCR detected B. microti in the blood and spleens of infected gerbils (Meriones unguiculatus) and also in laboratory-induced infections of I. ricinus ticks. B. microti DNA was detected by PCR in all pooled samples of nymphs and the majority of adults that had fed as larvae and nymphs, respectively, on gerbils with acute infection of the European isolate, confirming that I. ricinus could serve as a vector in Europe. The American isolate, GI, proved to be equally infective for larval and nymphal I. ricinus as the HK strain, despite a very different appearance in gerbil erythrocytes. Nymphs infected with the HK and GI strains readily infected gerbils. In contrast to the finding in acute infections, ticks that fed on gerbils with chronic infections of HK and GI did not become infected. It was also found that the HK strain was not transmitted transovarially. The finding that a B. microti strain (GI) from a distant geographical region (United States) can infect and be transmitted by I. ricinus suggests that other European B. microti strains, in addition to the HK strain used here, are probably infective for I. ricinus, supporting the view that infection of humans with European B. microti may be a regular occurrence. (+info)
Cloning of a truncated Babesia equi gene encoding an 82-kilodalton protein and its potential use in an enzyme-linked immunosorbent assay.
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To isolate Babesia equi genes encoding immunodominant proteins, a cDNA expression library prepared from B. equi mRNA was immunoscreened with B. equi-infected horse serum. Eighteen positive cDNA clones were obtained, and the clone that showed the strongest immunoreactivity, designated Be82, was further characterized. The Be82 gene consisted of 1,953 bp and contained a partial open reading frame lacking the 5'-terminal sequence. As shown by Western blot analyses, immune sera from mice intraperitoneally injected with the Be82 gene product recognized the 82- and 52-kDa proteins of B. equi but not those of Babesia caballi. The glutathione S-transferase fusion protein expressed in Escherichia coli that was purified and used as the antigen in the enzyme-linked immunosorbent assay reacted specifically with B. equi-infected horse sera. These results suggest that the Be82 gene product is a potential diagnostic antigen candidate in the detection of B. equi infection in horses that will be useful both in the performance of epidemiological studies and in the granting of quarantine passes. (+info)
The effect of macrophages on the erythrocyte oxidative damage and the pathogenesis of anemia in Babesia gibsoni-infected dogs with low parasitemia.
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The role of macrophages in the erythrocyte membrane oxidative damage and the pathogenesis of anemia in Babesia gibsoni-infected dogs with low parasitemia were investigated. Macrophages derived from peripheral blood monocytes (PBM) from B. gibsoni-infected dogs produced significantly higher chemiluminescent responses, indicating the release of reactive oxygen intermediates, than those from non-infected dogs when the cells were subjected to non-specific stimulation with phorbol 12-myristate 13-acetate (PMA) and opsonized zymosan (OZ), or infected dog erythrocyte membranes opsonized with infected dog serum. These results indicate that PBM of B. gibsoni-infected dogs with low parasitemia were highly activated compared to those of non-infected dogs. Furthermore, the membrane lipid peroxidation of normal dog erythrocytes incubated with PBM from B. gibsoni-infected dogs was significantly higher (p<0.05) than that of erythrocytes incubated with PBM from non-infected dogs when the PBM were stimulated with the opsonized membranes. These results suggest that the oxidative damage of erythrocytes observed in B. gibsoni-infected dogs with low parasitemia might be induced, in part, by reactive oxygen species released from the activated PBM. On the other hand, the present study also showed a significant increase (p<0.001) of IgG-bound erythrocytes in B. gibsoni-infected dogs compared with such erythrocytes in non-infected dogs. The increase of IgG-bound erythrocytes in infected dogs might reflect the increase of erythrocytes with oxidative damage induced by the infection with B. gibsoni. The results of the present study suggest that the increase of IgG-bound erythrocytes in the circulation of infected dogs induce a high degree of erythrocyte loss via immunological phagocytosis by activated macrophages, resulting in severe anemia in spite of low parasitemia. (+info)
Seroepidemiologic studies on Babesia caballi and Babesia equi infections in Japan.
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Antibodies to Babesia caballi and Babesia equi were examined on a total of 2,019 horse serum samples that had been collected in 1971-1973 by the National Institute of Animal Health by enzyme-linked immunosorbent assay (ELISA) using recombinant proteins and by Western-blot analysis. Based on the criterion for positivity by ELISA, 5.4% (109/2,019) and 2.2% (44/2,019) had antibodies against B. caballi and B. equi, respectively. The ELISA-positive sera were further examined by Western blot; 30/109 for B. caballi and 2/ 44 for B. equi were positive for native B. caballi or B. equi, but none of them was seropositive for both infections. Based on the results of this study, further investigations should be required to survey horses that have arrived in Japan relatively recently and tick vectors of equine Babesia using ELISA with some recombinant protein, a parasite detection method in an in vitro culture of equine Babesia, and PCR testing. (+info)
Coexistence DNA of Borrelia burgdorferi sensu lato and Babesia microti in Ixodes ricinus ticks from north-western Poland.
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The tick Ixodes ricinus may carry microorganisms which cause serious human and animal diseases, i.a., the Lyme disease (borreliosis), caused by the spirochaete Borrelia burgdorferi sensu lato and babesiosis, induced by the protozoan Babesia microti. Both microbe species may co-occur in the same and other species of the genus tick and produce a mixed infection in humans and animals. The major objective of the study was to identify DNA of B. burgdorferi and B. microti in the I. ricinus ticks collected in spring and autumn 1999 from 6 sites in north-western Poland. The microbial DNA was identified with polymerase chain reaction (PCR). The marker used to detect the B. burgdorferi s.l. DNA was a fragment of the fla gene encoding the protein flagellin, while the B. microti DNA was detected with a fragment of the gene encoding 16S rRNA. A total of 550, 1,160, and 385 tick adults, nymphs, and larvae, respectively, were examined. Among the 155 (7.4%) B. burgdorferi- infected ticks and the 130 (6.2%) infected with B. microti, mixed infection was detected in 0.6% of individuals. The prevalence of coinfection differed between the tick developmental stages. Coinfection was most prevalent (3.1%) in females, males and nymphs being less affected (0.4 and 0.2%, respectively). No coinfection was revealed in the tick larvae. The study described was the first of its kind to be conducted in the former District of Szczecin. For the phenomenon of microbial co-occurrence and related mixed infections to be properly evaluated, the research will be continued. (+info)
Seroprevalence of Babesia infections in humans exposed to ticks in midwestern Germany.
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Babesiosis is considered to be an emerging tick-borne disease in humans worldwide. However, most studies on the epidemiology of human babesiosis to date have been carried out in North America, and there is little knowledge on the prevalence of infection and frequency of disease in other areas. The aim of this study was to investigate the prevalence of Babesia infections in a human population in Germany. A total of 467 sera collected between May and October 1999 from individuals living in the Rhein-Main area were tested for the presence of immunoglobulin G (IgG) and IgM antibodies to antigens of Babesia microti and Babesia divergens by indirect fluorescent-antibody (IFA) tests. These sera were derived from 84 Lyme borreliosis patients suffering from erythema migrans, 60 asymptomatic individuals with positive borreliosis serology, and 81 individuals with a history of tick bite. Cutoff values for discrimination between seronegative and seropositive results in the IFA tests were determined using sera from 120 healthy blood donors and 122 patients suffering from conditions other than tick-borne diseases (malaria, n = 40; toxoplasmosis, n = 22; syphilis, n = 20; Epstein-Barr virus infection, n = 20; and presence of antinuclear antibodies, n = 20). The overall specificities of the IFA tests for B. microti and B. divergens were estimated to be >or=97.5%. Positive IgG reactivity against B. microti antigen (titer, >or=1:64) or B. divergens antigen (titer, >or=1:128) was detected significantly more often (P < 0.05) in the group of patients exposed to ticks (26 of 225 individuals; 11.5%) than in the group of healthy blood donors (2 of 120 individuals; 1.7%). IgG antibody titers of >or=1:256 against at least one of the babesial antigens were found significantly more often (P < 0.05) in patients exposed to ticks (9 of 225) than in the control groups (1 of 242). In the human population investigated here, the overall seroprevalences for B. microti and B. divergens were 5.4% (25 of 467) and 3.6% (17 of 467), respectively. The results obtained here provide evidence for concurrent infections with Borrelia burgdorferi and Babesia species in humans exposed to ticks in midwestern Germany. They also suggest that infections with Babesia species in the German human population are more frequent than believed previously and should be considered in the differential diagnosis of febrile illness occurring after exposure to ticks or blood transfusions, in particular in immunocompromised patients. (+info)
Entomologic and serologic evidence of zoonotic transmission of Babesia microti, eastern Switzerland.
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We evaluated human risk for infection with Babesia microti at a site in eastern Switzerland where several B. microti-infected nymphal Ixodes ricinus ticks had been found. DNA from pooled nymphal ticks amplified by polymerase chain reaction was highly homologous to published B. microti sequences. More ticks carried babesial infection in the lower portion of the rectangular 0.7-ha grid than in the upper (11% vs. 0.8%). In addition, we measured seroprevalence of immunoglobulin (Ig) G antibodies against B. microti antigen in nearby residents. Serum from 1.5% of the 396 human residents of the region reacted to B. microti antigen (>1:64), as determined by indirect immunofluorescence assay (IgG). These observations constitute the first report demonstrating B. microti in a human-biting vector, associated with evidence of human exposure to this agent in a European site. (+info)