A Polymerase Chain Reaction (PCR) and Southern blot hybridization for the detection of Theileria annulata are described. The PCR used primers amplifying a 785 base-pair fragment of the T. annulata gene which encodes the 30 kDa major merozoite surface antigen, Tams1. The sensitivity of the PCR in bovine blood was 1 piroplasm in 1 microl of blood. T. buffeli, T. parva, Babesia bigemina, B. bovis and B. divergens were not detected. The PCR detected down to 1 infected acinus/tick in resting and partially fed adult Hyalomma anatolicum anatolicum ticks and was negative for T. lestoquardi and T. equi, which are transmitted by this tick but are not infective to cattle. The specificity of the PCR was checked using 30 stocks of T. annulata, all of which were detected. Three stocks of T. lestoquardi, 4 of T. equi and 1 each of T. buffeli, T. parva, B. bigemina, B. bovis and B. divergens were used to ascertain there were no cross-reactions. A nested PCR using separate primers for the first reaction and the same primers for the second reaction detected T. annulata to the same sensitivity and specificity in saponin-extracted DNA samples stored for long periods at -20 degrees C.
Brucellosis remains an animal and public health concern in South Africa, given the intensity and widespread distribution of outbreaks in cattle. We conducted a cross-sectional survey among cattle keepers in the Whittlesea community of the Eastern Cape Province of South Africa, which utilises communal grazing. Individual cattle keepers (
N
= 227) who attended prearranged meetings in selected villages were interviewed using a structured questionnaire to assess their knowledge, attitude and practices (KAP) regarding bovine brucellosis. We compared KAP scores between previous brucellosis-affected villages and unaffected villages. We compared attitude and practices scores between those who had heard of brucellosis and those who had not and between those above the 75th percentile knowledge score and those below. The KAP for the study population were described using frequency tables. Scores of different groups were compared using the Welch
t
-test or the Wilcoxon rank-sum test. Knowledge scores of those who had heard of brucellosis (60%) showed a bimodal distribution with a 0/18 primary peak and 5–6/18 secondary peak. Attitude scores showed a median of 7/14 (interquartile range [IQR] 6–9), with 98% requesting more information on brucellosis. Practices scores showed a median of 6/18 (IQR 3–8), with high-risk practices identified that could facilitate brucellosis transmission. There were significant differences in attitude and practices scores between the groups above and below the 75th percentile knowledge score. The community showed poor knowledge, poor to average practices and average to good attitude. Identified high-risk practices highlight the risk of potential introduction and transmission of brucellosis between cattle and zoonotic transmission to humans.
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