Attempts to transmit bovine anaplasmosis with Anopheles quadrimaculatus Say (Diptera: Culicidae) and Dermacentor andersoni Stiles (Acarina: Ixodidae).
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This study investigated the seroprevalence of antibodies against Anaplasma phagocytophilum in Berlin/Brandenburg, north-eastern Germany. During 1994-2001, 422 sera from patients with proven tick-exposure (specimens with antibodies against Borrelia burgdorferi) were compared with 249 control sera. Using indirect fluorescent antibody testing, significantly more positive samples were detected among Borrelia antibody-positive specimens (4.5%, 95% CI 2.5-6.5%) than among controls (1.2%, 95% CI 0.5-1.9%; p < 0.05). While six (2.2%, 95% CI 1.3-3.1%) samples were positive among Borrelia antibody-positive sera between 1994 and 1997, 13 (8.7%, 95% CI 6.9-10.5%) were positive between 1998 and 2001 (p < 0.01), indicating an uneven annual seroprevalence.
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A multiplex PCR/DNA probe assay was used to monitor Babesia bovis, B. bigemina and Anaplasma marginale infection in cattle introduced to a Boophilus microplus-infested area in Veracruz, Mexico. Eight intact, 18-month-old, cross-bred beef cattle (four naive, Group A; four Babesia species--premunized, Group B) were immediately exposed to ticks after arrival and were clinically monitored from day 6 to day 98 post-exposure (PE) to ticks. Blood sample analysis for DNA detection by the MPCR/DNA probe assay showed that Group A animals were infected with B. bovis from day 11 up to day 22 PE, requiring treatment on days 17-20. Group B animals were detected positive to B. bovis on days 17-20, did not require treatment and remained persistently infected from days 70 to 84 PE. Treatment of Group A animals delayed the infection with B. bigemina. These animals became positive to the parasite on days 63-77 PE. In contrast, Group B animals (untreated) showed B. bigemina infection on days 21-26 and 63-84 PE. One animal was positive for A. marginale infection on days 63-66 PE, the rest of the animals became so on days 80-98 PE. All infected animals required treatment with oxytetracycline. Monitoring the triple hemoparasite infection with the MPCR/DNA probe assay provided important epidemiological information. Thus, precautionary measures can be established when cattle are moved to a babesiosis/anaplasmosis risk area.
Anaplasma marginale was propagated in a continuous tick cell line and detergent-solubilized infected cells were used as antigen in a competitive ELISA (C-ELISA) for detection of Anaplasma-specific antibody in bovine sera. Positive control sera competed well (> or = 35% inhibition) with an A. marginale-specific monoclonal antibody for binding to this antigen, while negative sera failed to compete (< 35% inhibition). The C-ELISA was compared to the standard complement-fixation test (CFT) using 2,208 bovine sera. Overall, C-ELISA was more sensitive than CFT (24.9% versus 9.4%), mainly because CFT yielded "suspicious" or "anti-complementary" results in 10.5% of the sera and also failed to identify several vaccinated and carrier cattle that were C-ELISA-positive. The apparent agreement between CFT and C-ELISA was 89.6% and the kappa value was 0.6. These results show that this C-ELISA would be a suitable replacement of the CFT as the standard test for detection of A. marginale antibody.
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A capillary agglutination (CA), a complement fixation (CF), a plate agglutination (PT) and an indirect fluorescent antibody (IFA) test to detect humoral antibodies to Anaplasma marginale are described. Serums from 3, 4 or 5 groups of cattle were used to examine the efficiency of the tests. Agreement between all 4 tests was 86.6%. Agreement between pairs of tests was greater. The CF test was the most sensitive while the PT test was the least sensitive. However the PT could be carried out very rapidly and was suggested as the best screening test, providing improved antigen preparation techniques could increase sensitivity. The CA, CF and IFA tests all showed a stronger homologous antibody reaction when A. marginale antigen was tested against serums obtained from cattle infected with either A. marginale or A. centrale. Antibodies in the A. marginale serums were first detected by day 7 post-inoculation, rose to peak around day 29 and were still present on day 200. Antibodies in the A. centrale serums were first detected by day 29 rose to a peak around day 50 and had disappeared by day 150.
Direct fluorescent antibody (DFA) and Giemsa staining of Anaplasma marginale were compared in smears collected serially at post-mortem (PM) from 11 experimentally infected calves. Once smears had been prepared and air-dried they could be held for at least 5 days before staining with either technique with no noticeable change in staining quality. DFA staining was more sensitive in detecting anaplasms in smears than Giemsa staining. Anaplasma spp could be differentiated from Babesia bovis and B. bigemina by DFA staining but there were cross reactions between A. marginale and A. centrale. Blood smears prepared from subcutaneous vessels in the legs provided better diagnostic material than kidney, heart and lung smears. Brain smears were not suitable for PM diagnosis using either staining technique.
Comparative studies were carried out on the susceptibility of 8 Bos taurus and 10 predominantly Bos indicus cross cows aged 48 months to Anaplasma marginale infection. Measurements were carried out on bodyweight, parasitaemia, haematological factors, humoral antibody response and serum transaminase levels. Except for minor differences in reaction Bos indicus and Bos taurus animals were equally susceptible to infecton with the strain of A. marginale employed. Such differences observed were not significant.
Natural infection of calves with Anaplasma marginale was studied in two endemic areas of tropical north Queensland. Infection, assessed by a micro-complement fixation test, occurred throughout the year but was most frequent just after the summer wet season. Infection was clinically inapparent. Calves vaccinated with A. centrale did not show improved growth rate when compared with non-vaccinated calves. The complement fixation test was shown to be most effective in the detection of recent infections.
Bos indicus-crossbred calves exposed to natural Babesia bovis infection in wet and dry tropical environments in northern Queensland were tested for antibodies using the indirect haemagglutination (IHA) test. There was evidence of maternal antibodies suggesting endemicity of B. bovis in the 2 environments. Within one year 35.3 to 94.6% of the calves had acquired B. bovis infection. The highest titres and most rapid conversion rates were observed just after the wet season when the tick population was at its peak. Clinical disease was not observed. A third group of Bos indicus-cross cattle of varying ages in a dry tropical environment of north Queensland was examined for antibodies against B. bovis and Anaplasma marginale using the IHA and complement fixation (CF) tests, respectively. B. bovis and A. marginale antibodies were detected on the farm but clinical disease did not occur.
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Serologic and molecular evidence of Anaplasma phagocytophilum has been demonstrated in white-tailed deer (WTD; Odocoileus virginianus), and deer are an important host for the tick vector Ixodes scapularis. In this study, we describe experimental infection of WTD with A. phagocytophilum. We inoculated four WTD with a human isolate of A. phagocytophilum propagated in tick cells. Two additional deer served as negative controls. All inoculated deer developed antibodies (titers, > or =64) to A. phagocytophilum, as determined by an indirect fluorescent antibody test, between 14 and 24 days postinfection [p.i.]), and two deer maintained reciprocal titers of > or =64 through the end of the 66-day study. Although morulae were not observed in granulocytes and A. phagocytophilum was not reisolated via tick cell culture of blood, 16S reverse transcriptase nested PCR (RT-nPCR) results indicated that A. phagocytophilum circulated in peripheral blood of three deer through at least 17 days p.i. and was present in two deer at 38 days p.i. Femoral bone marrow from one deer was RT-nPCR positive for A. phagocytophilum at 66 days p.i. There was no indication of clinical disease. These data confirm that WTD are susceptible to infection with a human isolate of A. phagocytophilum and verify that WTD produce detectable antibodies upon exposure to the organism. Because adults are the predominant life stage of I. scapularis found on deer and because adult I. scapularis ticks do not transmit A. phagocytophilum transovarially, it is unlikely that WTD are a significant source of A. phagocytophilum for immature ticks even though deer have a high probability of natural infection. However, the susceptibility and immunologic response of WTD to A. phagocytophilum render them suitable candidates as natural sentinels for this zoonotic tick-borne organism.