This study was designed to investigate the prevalence of bovine anaplasmosis among cattle from various areas in Wassit governorate; the investigation was performed on 184 blood samples collected from suspected cattle suffering from fever (41°C), severe anemia, pale mucus membrane, progressive emaciation and drop in milk yield, including 85 male and 99 female cattle, aged from < 1 year to > 2 years, the samples were collected during the period of October 2012 - April 2013 from AL-Kut, AL-hayy, AL-Bashair, AL-Moufaqia and AL-Noamania areas to investigate antibodies against Anaplasma parasite by using indirect enzyme-linked immunosorbent assay (ELISA) test and to determine the species of genus Anaplasma by using RFLP-PCR technique and by also measuring some biochemical parameters to indicate the effect of the disease on liver function. The results of ELISA test showed that the rate of infection was 13.04%, the rate of infection was different between age groups and were 8, 11.25 and 16.45% in ages < 1, 1-2 and 2 - 3 years, respectively. The study revealed that females were given higher percent of infection 14.14% than males 11.7%, there is no significant differences under p > 0.05 according to age groups and sex. The highest rate of infection was recorded in AL-Kut, 17.14% followed by AL-hayy, 14% and AL-Bashair, 10% and the lowest rate was recorded in AL-Noamania, 8.33% and AL-Moufaqia, 5%; the study showed significant differences in incidence of disease between study districts and area in Wassit governorate at p > 0.05. The most sensitive method for the diagnosis of anaplasmosis is the method of polymerase chain reaction, DNA extraction was performed only on 24 blood samples which were positive for Anaplasma spp. by ELISA test, the extracted DNA from blood cells were analyzed by PCR and PCR-RFLP technique using primers derived from 16S rRNA gene and restriction endonuclease Bst1107I enzyme which can recognize the sequence (GTATAC) in corresponding PCR product of Anaplasma marginale and cut it in the position 68 and 509, whereas the used restriction enzyme cannot cut the corresponding PCR product of other Aanaplasma spp. and the result was 20 from 24 which were positive for Aanaplasma spp. by PCR and 18 from 20 was positive for A. marginale. The results of liver enzymes activity showed significant increase in serum AST, ALT, CK and TBIL level in infected cattle (96.8±0.97, 42±0.52, 406±2.06 and 0.95±0.24, respectively) as compared to the control (65.5±1.26, 21.4±0.45, 142±14.17 and 0.27±0.05, respectively).
Key words: Anaplasma marginale, indirect enzyme-linked immunosorbent assay (ELISA), RFLP-PCR, biochemical parameters.
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