Comparison of duplex PCR and microscopic techniques for the identification of Babesia bigemina and Babesia bovis in engorged female ticks of Boophilus microplus.
A single-step duplex polymerase chain reaction (PCR) technique and traditional microscopic examination of haemolymph smears were used to detect Babesia bigemina and/or Babesia bovis infection in engorged female ticks of Boophilus microplus recovered from calves raised in an endemic area of the State of Minas Gerais, Brazil. In the PCR amplification of tick-derived DNA, pairs of oligonucleotide primers specific for a 278-bp sequence from B. bigemina and for a 350-bp sequence from B. bovis were used conjointly. The microscopic examination of haemolymph revealed that 16.7% of the engorged ticks were infected with Babesia spp., although no significant differences (rho > 0.05) were found in the infection rate of ticks collected from calves of different age groups. PCR analysis showed that 77.8% of the engorged ticks whose haemolymph contained sporokinetes were infected with B. bigemina, 7.8% with B. bovis and 14.4% with both protozoan species. However, the PCR assay further revealed that, amongst the engorged female ticks whose haemolymph was apparently negative for the presence of sporokinetes, 15.6% were infected with B. bigemina, 2.2% with B. bovis and 10.0% with both species. The duplex PCR method is thus more efficient and sensitive than the microscopic assay and also permits facile identification of the protozoa species present in engorged female ticks.
Quintão-Silva
M G
MG
Unifenas, Departmento de Parasitologia, Belo Horizonte-MG, Brazil.
Melo
M N
MN
Ribeiro
M F B
MF
D000818:Animals; D000831:Animals, Newborn; D001403:Babesia / Q000302:isolation & purification*; D016793:Babesia bovis / Q000302:isolation & purification; D001404:Babesiosis / Q000175:diagnosis / Q000635:transmission / Q000662:veterinary*; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis; D016054:DNA, Protozoan / Q000032:analysis*; D005260:Female; D006801:Humans; D008853:Microscopy / Q000379:methods / Q000662:veterinary*; D008970:Molecular Weight; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D011634:Public Health; D012680:Sensitivity and Specificity; D013984:Tick Infestations / Q000662:veterinary; D013987:Ticks / Q000469:parasitology*; D015047:Zoonoses
D016054:DNA, Protozoan