Sub-atomic and biochemical investigation of Anaplasma marginale in dairy cattle in Wassit Province of Iraq

Abstract

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 ALNoamania 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).