Resumo Hemoplasmas são bactérias epieritrocíticas que infectam mamíferos. ‘Candidatus Mycoplasma haemoalbiventris’ foi detectado previamente em gambás-de-orelha-branca (Didelphis albiventris) das regiões sul e centro-oeste do Brasil. O presente estudo objetivou: i) triar os gambás para as doenças transmitidas por carrapatos (Piroplasmida e Anaplasmataceae); e ii) detectar e caracterizar as espécies de hemoplasma que infectam gambás nas cidades de Curitiba e Foz do Iguaçu, no Estado do Paraná, sul do Brasil. Trinta amostras de sangue de gambás-de-orelha-branca foram analisadas por PCR. Os animais não estavam infestados por ectoparasitos. O gene endógeno de mamífero gapdh foi amplificado em todas as amostras. Todos os gambás testaram negativos para Theileria/Babesia spp. e Ehrlichia/Anaplasma spp. por PCR, respectivamente, para os genes 18S rRNA e 16S rRNA. Uma PCR gene-específica, baseada no gene 16S rRNA de hemoplasmas, mostrou que três/13 (23,08%; CI 95%: 8,18-50,26%) gambás de Foz do Iguaçu foram positivos para Mycoplasma sp. hemotrópico. Todos os gambás de Curitiba testaram negativos para hemoplasmas. O sequenciamento de fragmentos dos genes 16S e 23S rRNA revelou que os animais estavam infectados pelo ‘Ca. M. haemoalbiventris’. Embora ‘Ca. M. haemoalbiventris’ seja prevalente em gambás no Brasil, os sinais clínicos associados à infecção e os prováveis vetores permanecem desconhecidos.

Abstract Hemoplasmas are epierythrocytic bacteria that infect mammals. ‘Candidatus Mycoplasma haemoalbiventris’ was detected in white-eared opossums (Didelphis albiventris) from southern and central-western Brazil. The present study aimed at: i) screening opossums for tick-borne (TBP) pathogens (Piroplasmida and Anaplasmataceae) and ii) detecting and characterizing hemoplasma species infecting opossums from Curitiba and Foz do Iguaçu cities in the Paraná State, southern Brazil. Thirty blood samples from white-eared opossums were evaluated by PCR assays. Animals were not infested by ectoparasites. The mammalian endogenous gapdh gene was consistently amplified in all samples. All opossums tested negative for Theileria/Babesia spp. and Ehrlichia/Anaplasma spp. by PCR based on 18S rRNA and 16S rRNA genes, respectively. A genus-specific PCR assay based on the 16S rRNA gene of hemoplasmas showed that three/13 (23.08%; CI 95%: 8.18-50.26%) opossums from Foz do Iguaçu were positive for hemotropic Mycoplasma sp. All opossums from Curitiba tested negative for hemoplasmas. Sequencing of both the 16S and 23S rRNA genes revealed that the animals were infected by ‘Ca. M. haemoalbiventris’. Although ‘Ca. M. haemoalbiventris’ is prevalent in opossums in Brazil, clinical signs associated with its infection and its putative vectors remain unknown.

The genetic variability of populations of Aedes (Stegomyia) aegypti (Linnaeus, 1762) is often low due to the intense selection pressure caused by chemical control measures. In this study, we evaluated the susceptibility of larvae and adults of this mosquito to chemical insecticides, the frequency of the Val1016IIe mutation, and the genetic variability of the mitochondrial ND4 gene fragment in the urban area of Foz do Iguaçu, Paraná. The populations of A. aegypti in the southern and central regions of the city were resistant to the diagnostic dose of temephos 0.0162 ppm. Additionally, we detected difference in susceptibility in the northern population. The resistance ratios (RR95) were 3.8, 4.1, and 4.6 for the populations in the north, central, and south of the city, respectively. The entire population of A. aegypti in Foz do Iguaçu is resistant to pyrethroids. The mortality rates are as follows: 72.2% for cypermethrin (CD: 146 mg i.a./m²) and 57% for deltamethrin (CD: 18 mg i.a./m²). These results corroborate the hypothesis that the mutated 1016Ile allele is present in all analyzed strata. Out of the 234 samples genotyped for the Val1016Ile mutation, 15% were homozygous dominant for the wild allele (Val/Val), 62% were heterozygous (Va/Ile) and 23% were homozygous for the recessive mutation (Ile/Ile). With respect to the genetic variability of the mitochondrial ND4 gene fragment, 93% of the sequences analyzed belonged to haplotype 1, and 7% belonged to haplotype 2. The genetic diversity was low, the fixation index was not significant, and gene flow was high. The control of A. aegypti in Foz do Iguaçu using temephos and pyrethroids may be compromised because of the reduced vector susceptibility. Populations of A. aegypti, that undergoes a sudden reduction in effective population size and become resistant to pyrethroids may differ from the original population in vector capacity.