Claudia María Ríos-Velásquez

The Amazon Basin harbors a high diversity of phlebotomine sand flies, including several species that act as vectors of zoonotic pathogens such as Leishmania. Land-use changes, particularly forest conversion to agriculture, alter the sand fly diversity and community structure, with implications for the transmission of American Tegumentary Leishmaniasis (ATL). We evaluated the effects of forest-to-agriculture conversion on sand fly diversity and species composition in two rural areas, on opposite sides of the Amazonas River, in the Brazilian Amazon Region. Sand flies were collected using Center of Disease Control (CDC) light traps, installed in the forest and cropland environments, at the Presidente Figueiredo (North) and Urucurituba (South) municipalities, in Amazonas, Brazil. We collected a total of 1778 phlebotomine sand flies from 15 genera and 69 species. The most abundant species were Micropygomya rorotaensis (n = 436; 24.52%), Nyssomyia antunesi (n = 297; 16.70%), Sciopemyia sordellii (n = 101; 5.68%), Bichromomyia flaviscutellata (n = 84; 4.72%) and Evandromyia monstruosa (n = 72; 4.04%). In addition, four sand fly species were recorded for the first time in the Amazonas state: Brumptomyia mesai, Pressatia calcarata, Evandromyia aldafalcaoae and Lutzomyia carvalhoi. Sand fly richness, diversity, and community composition varied between riversides and environments, reflecting strong effects of anthropogenic disturbance. Although croplands supported reduced and more heterogeneous assemblages, several medically important vector species persisted across both environments. Species turnover was high, but patterns of species loss were weak, suggesting that community reorganization was driven by non-directional compositional change process. Our results indicate that land-use change reshapes sand fly communities without eliminating disease vectors, potentially increasing ATL transmission risk at the forest–anthropic interface.

Visceral leishmaniasis is a zoonotic disease of public health importance in Brazil, with expansion into peri-urban areas. This cross-sectional study evaluated 450 dogs in a municipality of the Western Amazon, Brazil, using TR-DPP® serology, ELISA confirmation, hematological analysis, PCR targeting the Hsp70 gene, and entomological surveys with CDC light traps. A total of 45 dogs (10%) were seroreactive, and 34 were confirmed by ELISA. Hemoparasites such as Babesia spp. were detected. All pooled samples were negative by PCR. Six species of phlebotomine sand flies were identified, including Nyssomyia antunesi and Migonemyia migonei. The findings suggest early exposure of dogs in an area of emerging epidemiological risk, highlighting the importance of integrated surveillance strategies.

Malaria remains a public health issue across the world. Different methods have been analyzed to achieve this disease’s elimination, such as the vector control of Anopheles spp. Control strategies include the use of different classes of insecticides, although the accelerated evolution of vectors resistant to them makes the development of alternative control methods necessary. Therefore, entomopathogenic fungi have been considered to be promising biopesticides, given that they are safe for human beings and the environment. This study aimed to evaluate the entomopathogenic activity of fungi collected in the Amazon Rainforest against adult female Anopheles aquasalis mosquitoes. Females were exposed to four different species of fungi and observed daily to evaluate their survival rate. Also, fungi species’ behavior was analyzed through scanning electron microscopy (SEM). Those exposed to Trichoderma harzianum and Penicillium citrinum had their survival rate reduced. SEM confirmed the development of fungi on the mosquitoes after 48 h. The findings suggest that the entomopathogenic potential of the fungi used in this study should be considered, given the reduction in the survival rate of Anopheles aquasalis mosquitoes.

The northern region of Brazil is endemic for American Tegumentary Leishmaniasis (ATL) primarily caused by Leishmania guyanensis and transmitted by the sand fly Nyssomyia umbratilis. The disease occurs at different rates in the municipalities of Manacapuru (MAN) and Rio Preto da Eva (RPE), located in the state of Amazonas. Despite their geographic proximity and separation by the Rio Negro, MAN has a low incidence, whereas RPE reports a significantly higher number of cases. Since the vector is present in both locations, potential biological differences in N. umbratilis may influence transmission. Previous studies suggested genotypic and phenotypic differences in N. umbratilis from both localities. To investigate the molecular factors underlying their potentially differential vectorial capacities, we performed a comparative proteomic analysis of dissected insect intestines from both localities. Our results revealed that sand flies from MAN showed a higher abundance of proteins related to gene transcription, protein translation, amino acid and proton transport, innate immune response and intestinal motility. Since the importance of microbiota has previously been shown in parasite–vector interactions, we also identified bacteria from both vector populations. We detected bacteria specific to each population and, exclusively in MAN, some species described in the literature as having parasiticidal properties. These findings highlight molecular and microbial peculiarities that could contribute to the observed difference in ATL prevalence in the two areas.

Trichomitus batrachorum is a species of trichomonad that has gained attention due to its ecological importance and potential interactions with various hosts, such as amphibians (anurans) and reptiles (lizards and chelonians), where it has been recorded in the gastrointestinal tract of these vertebrates, specifically in their feces. Molecular studies have placed this flagellated protist within the Metamonada clade. Unlike parabasalids that inhabit endothermic mammals in relatively stable temperature conditions, protists associated with ectothermic reptiles are subject to significant temperature fluctuations. The ability of T. batrachorum to thrive in the variable temperatures encountered by reptiles suggests that its parasitism may remain largely unaffected by climate change. In our study, we detected and characterized T. batrachorum from the liver tissue of the lizard species Ameiva ameiva, collected in Presidente Figueiredo Municipality, Amazonas State, Brazil. The identification of T. batrachorum was confirmed by cultivation technique, light microscopy, scanning electron microscopy and transmission electron microscopy for ultrastructural analyses, and sequencing the 5.8S rDNA (region ITS1- ITS2) and 18S rRNA (ribosomal RNA) genes. One potential interpretation for this finding is that the flagellates may have migrated from the intestine to the bile duct, ultimately reaching the liver. This is the first successful characterization of T. batrachorum in the liver of a lizard, and provides a solid foundation for further research to elucidate the potential pathogenicity of this flagellate and the role of A. ameiva in the epidemiology of parabasalids in other animal species.

Malaria continues to affect millions of people around the world. Current control strategies have not been sufficient, and additional tools are required for malaria elimination. A promising approach is the use of bacteria from the mosquito microbiota, such as the Su_YN1 Serratia ureilytica bacterium, that was shown to strongly impair development of P. falciparum and P. berghei in Anopheles mosquitoes. To evaluate the potential of the S. ureilytica Su_YN1 bacterium as a blocker of the Plasmodium parasite in South American vectors, we investigated its effects on mosquito fitness and survival. We found that this bacterium does not affect longevity, blood feeding, fecundity and fertility of Anopheles aquasalis, an important South American vector. Overall, our results provide baseline support for the potential implementation of Su_YN1 for the control malaria transmission in South America.

In the fight against malaria, the key is early treatment with antimalarial chemotherapy, such as artemisinin-based combination treatments (ACTs). However, Plasmodium has acquired multidrug resistance, including the emergence of P. falciparum strains with resistance to ACT. The development of novel antimalarial molecules, that are capable of interfering in the asexual and sexual blood stages, is important to slow down the transmission in endemic areas. In this work, we studied the ability of the mettalo copper-cinchonine complex to interfere in the sexual and asexual stages of Plasmodium. The tested compound in the in vitro assay was a cinchonine derivative, named CinCu (Bis[Cinchoninium Tetrachlorocuprate(II)]trihydrate). Its biological functions were assessed by antiplasmodial activity in vitro against chloroquine-resistant P. falciparum W2 strain. The mice model of P. berghei ANKA infection was used to analyze the antimalarial activity of CinCu and chloroquine and their acute toxicity. The oocyst formation-blocking assay was performed by experimental infection of Anopheles aquasalis with P. vivax infected blood, which was treated with different concentrations of CinCu, cinchonine, and primaquine. We found that CinCu was able to suppress as high as 81.58% of parasitemia in vitro, being considered a molecule with high antiplasmodial activity and low toxicity. The in vivo analysis showed that CinCu suppressed parasitemia at 34% up to 87.19%, being a partially active molecule against the blood-stage forms of P. berghei ANKA, without inducing severe clinical signs in the treated groups. The transmission-blocking assay revealed that both cinchonine and primaquine were able to reduce the infection intensity of P. vivax in A. aquasalis, leading to a decrease in the number of oocysts recovered from the mosquitoes’ midgut. Regarding the effect of CinCu, the copper-complex was not able to induce inhibition of P. vivax infection; however, it was able to induce an important reduction in the intensity of oocyst formation by about 2.4 times. It is plausible that the metallo-compound also be able to interfere with the differentiation of parasite stages and/or ookinete-secreted chitinase into the peritrophic matrix of mosquitoes, promoting a reduction in the number of oocysts formed. Taken together, the results suggest that this compound is promising as a prototype for the development of new antimalarial drugs. Furthermore, our study can draw a new pathway for repositioning already-known antimalarial drugs by editing their chemical structure to improve the antimalarial activity against the asexual and sexual stages of the parasite.

Anopheles darlingi is the main malarial vector in the Brazilian Amazon region. An. nuneztovari s.l., An. triannulatus s.l., An. evansae, and An. benarrochi s.l. do not have a defined role as malarial vectors, although they have been found to be naturally infected with Plasmodium vivax, and some develop oocysts. In this study, we evaluated the importance of low numbers of oocysts in sporozoite salivary gland invasion and transmission. Field-collected mosquitoes were experimentally infected with P. vivax. The infection rates and oocyst and sporozoite infection intensities were evaluated and compared with those of An. aquasalis. We found the highest number of oocysts in An. darlingi (mean = 39.47) and the lowest in An. nuneztovari s.l. (mean = 2). The highest number of sporozoites was observed in An. darlingi (mean = 610) and lowest in An. benarrochi s.l. (mean = 30). Plasmodium vivax DNA was detected in the saliva of all mosquito species after a blood meal. Regardless of the number of oocysts, all species transmitted sporozoites during blood meals. Considering the abundance of these mosquitoes and transmission of sporozoites, it is logical to assume that An. nuneztovari s.l. and An. triannulatus s.l. are involved in the transmission of P. vivax.

Three species of Corethrella Coquillett, 1902 from the state of Amazonas, Brazil are described as new to science based on female adult specimens. Corethrella cabocla Feijó, Belchior, Marialva & Pessoa sp. nov. possesses four large setae on the frons between the ventromedial area of ommatidia, a wide clypeus with 1–4 setae, a wing with the apex of R2 basal to the apex of M2 and with a midlength band, and with the abdomen entirely dark brown. Corethrella ielemdei Feijó, Ramires, Lima & Pessoa sp. nov. possesses an elongated coronal suture, four large setae on the frons between the ventromedial area of ommatidia, a clypeus squarish with 42–43 setae, a wing with the apex of R2 basal to the apex of M1 and with a midlength band and dark scales on the basal and subbasal areas of the anterior margin, legs with dark scales, and with the abdomen entirely dark brown. Corethrella menini Feijó, Picelli, Ríos-Velásquez & Pessoa sp. nov. possesses wings with the apex of R2 basal to the apex of M2 and a midlength band, with darker basal scales along all veins, basal band dark scales on C, Sc, R, M, and Cu and the abdomen entirely dark brown. With the addition of the new species, the numbers of frog-biting midges described in the Amazon basin, Brazil and in Neotropical region are now 31, 49 and 80 species, respectively.