Milton Hissashi Yamamura

Detection of Anaplasma marginale DNA in Larvae of Boophilus microplus Ticks by Polymerase Chain Reaction

Abstract: Boophilus microplus larvae from two different sources were used for the detection of Anaplasma marginale DNA: larvae A, which were collected from a pasture of an endemic farm, and larvae B, which originated from engorged female ticks fed on calves with no clinical signs of disease and with low rickettsemia (approximately 0.01 to 1.0%). Larvae A were collected monthly, from January to May in 2001. Two hundred engorged female ticks fed on calves that provided larvae B were divided into groups of 10 and kept in a controlled environment at either 18°C or 28°C. Fifty larvae were used from each sample for DNA extraction, and 5 μL of DNA were submitted to amplification of the sequence of msp5 gene of A. marginale by polymerase chain reaction (PCR). Seven out of 50 samples of larvae A (14%) were positive for the presence of DNA of A. marginale showing amplified product of 457 bp. Ten out of 91 samples of larvae B (11%) kept at 18°C were positive, and all larvae B at 28°C were negative. Thus, this study confirmed the presence of A. marginale DNA in B. microplus larvae by PCR. The EcoRI restriction enzyme analysis confirmed the specificity of the amplicon, which resulted in two fragments: 265 bp and 192 bp. The sequencing analysis of the amplicon from larvae demonstrated 98% homology with the msp5 sequence from Florida A. marginale strain.

Occurrence of gastrointestinal and renal helminths in Zenaida auriculata (Des Murs, 1847) trap-captured from Brazil

This study aimed to determine the prevalence of gastrointestinal and renal helminths from naturally infected Zenaida auriculata captured in Londrina, Paraná State. Two hundred and one Eared doves were trapped and the gastrointestinal and renal helminths were collected and identified according to morphological structures. One hundred and sixteen (57.71%) doves were parasitized by helminths with specific prevalences for Ornithostrongylus quadriradiatus in 50 doves (24.88%), Ascaridia columbae in 47 (23.38%), Paratanaisia bragai and P. confusa in 34 (16.92%), Tetrameres fissispina in 17 (8.46%), Synhimantus nasuta in 14 (6.47%), Brachylaima mazzantii in 4 (1.99%) and Raillietina allomyodes in 2 doves (1.00%). Seventy four/201 (37.00%) birds were infected with only one species, and 96/201 (48.00%) pigeons were infected with nematodes. The association between different classes of helminths occurred in 40/201 (20.00%) animals. The results showed statistically differences between the presence of nematode (p = 0.00001) and trematode species (p ≤ 0.05) in the doves, and there was an association between the local of capture and the presence of trematodes and A. columbae (p ≤ 0.05). This study is the first to report the infection of Z. auriculata from Brazil with O. quadriradiatus, A. columbae, T. fissispina, S. nasuta, R. allomyodes, P. bragai and P. confusa.