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This study aimed to optimize molecular methods for detecting DNA of Hepatozoon spp. as well as identify the phylogenetic relationships of Hepatozoon strains naturally infecting domestic dogs in Belém, Pará, northern Brazil. Blood samples were collected from 138 dogs, and screened for Hepatozoon spp. using a new nested PCR assay. Positive samples were subjected to genetic characterization based on amplification and sequencing of approximately 670bp of the Hepatozoon spp. 18S rRNA. Of the positive dogs, four shared the haplotype Belém 01, one dog presented the haplotype Belém 02 and two dogs shared the haplotype Belém 03. A Bayesian inference indicates that haplotypes Belém 01 and Belém 02 are phylogenetically related to H. canis, while Belém 03 is related to H. americanum. Overall, based on the first molecular evidence of H. americanum in Brazilian domestic dogs, the proposed protocol may improve the epidemiological investigation of canine hepatozoonosis., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Babesiosis is an infectious hemolytic disease that occurs worldwide, and is caused by a protozoan of the Babesia genus (Apicomplexa). Little is known about this genus; therefore, this study conducted a molecular characterization of Babesia spp in naturally infected dogs in northern Brazil. Blood samples were collected from 172 dogs from metropolitan Belém, and screened for Babesia spp using semi-nested polymerase chain reactions. Subsequently, 18S rDNA gene fragments were amplified and sequenced. Alignments of the 27 nucleotide sequences returned fragments measuring 1023 bp in size, which did not include any polymorphic sites (100% identity). This genotype was very similar to the 18S rDNA gene in B. vogeli. This study provides an important molecular characterization of the parasite responsible for canine babesiosis in naturally infected dogs in metropolitan Belém. In addition, the 18S rDNA gene in B. vogeli may represent the most common genotype occurring in South America.

Babesiosis is a hemolytic disease caused by protozoans of the genus Babesia (Apicomplexa). This disease occurs worldwide and is transmitted by ticks to a variety of mammals, including humans. The objective of the present study was to optimize a molecular approach for the detection of a fragment of 18S rDNA of Babesia canis, Babesia vogeli, Babesia rossi or Babesia gibsoni based on a single semi-nested Polymerase Chain Reaction (PCR), and compare the efficiency of this approach with that of a simple PCR protocol. To this end, 100 blood samples collected from dogs with suspected hemoparasite infections were analyzed. A comparison of the results of simple PCR and semi-nested PCR indicated a highly significant difference (p value = 0.0000). While only five (5%) of the samples tested positive using the simple protocol, 22 (22%) were positive using the snPCR technique. The results of this study reinforce the findings of previous studies, which have demonstrated the greater sensitivity of tests based on nested or semi-nested PCR. Therefore, to avoid false-negative results due to low levels of parasitemia, we suggest the preferential use of this protocol in epidemiological studies of canine babesiosis, particularly those that require reliable estimates of the prevalence of infection.