Search Results (53)

Information on Culicoides transmitted parasites, especially trypanosomatids, infecting animals and insects, is scarce. Our goal was to clarify the seasonal patterns of both Culicoides and trypanosomatids detected in these insects and the relationships between Culicoides abundance and meteorological parameters. UV light traps were used to collect biting midges in four study sites in 2022–2023; collected Culicoides females were dissected and analyzed using microscopy and PCR-based methods. Out of 1631 parous Culicoides females belonging to 14 different species, 6.5% were found to be infected with trypanosomatids (5.0% with at least three Trypanosoma species and 1.5% with monoxenous parasites). The highest Culicoides abundance was detected in June. The prevalence of trypanosomatids in biting midges increased during the summer (5.3% in June, 8.8% in July, and 11.2% in August). Temperature was recorded to be a presiding environmental gradient structuring Culicoides species composition, while wind speed and precipitation explained little of the variation. Our results indicate that both avian and mammalian trypanosomes can be found in these insects, although further research is needed to better understand the development of these parasites in biting midges and Culicoides vectorial capacity. Full article

Passerine birds (n = 3335) of 19 species were caught and investigated for the presence of Trypanosomatidae and Onchocercidae parasites using the buffy coat method, microscopy and PCR in Ventės Ragas, Lithuania. Data on the spread patterns of these parasites are still lacking. The prevalences of Trypanosoma parasites in birds of different species varied from 2.2% to 36.1%, while the prevalences of Onchocercidae parasites varied from 0% to 17.3%. Statistically significant differences between spring and autumn in the prevalences of Trypanosomatidae were determined for Acrocephalus schoenobaenus, Hirundo rustica and Turdus philomelos. No significant differences between the prevalences of Onchocercidae in spring and autumn were determined. The prevalence of Trypanosoma was significantly higher for long-distance migrant birds compared with short-distance migrants, for omnivorous birds compared with insectivorous birds, and for open-nesting birds compared with birds nesting in nest boxes. The prevalences of Onchocercidae parasites did not differ for the same bird groups except for the prevalence in omnivorous birds, which was higher compared with insectivorous birds. Both groups of parasites were detected in juveniles, showing the presence of transmission in the study area. The diet, breeding behaviour and migration features of avian hosts can influence the prevalence of avian blood parasites. Full article

Trypanosoma cruzi, the causal agent of Chagas disease, exhibits great genetic diversity, which has been related to its biological properties. However, these are poorly known in strains from the endemic area of Hidalgo. To assess the parasite’s virulence, we evaluated parasitemia, mortality, and tropism in thirteen organs of CD1 mice during the acute phase of infection. For genotyping, we amplified the mini-exon gene from T. cruzi DNA using PCR. All five isolates were identified as belonging to DTU TcI. The peak of parasitemia occurred between 25 and 29 days post-infection. The Tultitlán and Olma isolates did not cause any mouse deaths, whereas Ixcatépec produced 100% mortality. Mice infected with the Barrio Hondo isolate exhibited the highest parasitemia, while those infected with Cuatecomaco had the lowest. The five isolates generated varying degrees of infection and chronic inflammation; only two isolates triggered acute pancreatitis and myocarditis. No amastigote nests were found in the hearts of mice infected with the Ixcatépec isolate. Our findings suggest that the damage caused by T. cruzi strains from Hidalgo may extend beyond cardiac lesions in the acute phase of Chagas disease regardless of their classification as TcI and variability in parasitemia levels. Full article

Leishmaniasis and Chagas disease are parasitic diseases considered to be among the most important neglected diseases, with implications for both developed and developing countries. Currently, there are no effective therapeutic treatments for these diseases due to challenges in drug administration, high toxicity, high costs, and drug resistance. In this study, a series of eleven thymol derivatives were designed, synthesized, and evaluated for their in vitro kinetoplastid activity against Leishmania amazonensis and Trypanosoma cruzi, as well as their cytotoxicity against a murine macrophage cell line. The most active compounds, thymol anysoate (9) and thymol picolinate (10), displayed the highest kinetoplastid activity with IC₅₀ values of 22.87 and 25.16 µM against L. amazonensis and T. cruzi, respectively. Notably, both compounds demonstrated an excellent selectivity index against the mammal cell line. Structure–activity relationship studies revealed that the ester group plays a crucial role in activity. The most promising derivatives, 9 and 10, activate autophagy and apoptosis-like processes in the treated cells. Atomic force microscopy observations showed that derivative 9 induces the formation of cytoplasmic vacuoles, indicating an autophagic process, and drug-likeness analysis revealed that it meets all the pharmacokinetic criteria. Overall, these results highlight derivative 9 as a potential lead compound for the development of new drugs for the treatment of Trypanosomatidae infections and warrants further studies to elucidate the cell death cascade involved. Full article

Background/Objectives: Many tropical diseases such as malaria, Chagas, human African Trypanosomiasis, and Lymphatic filariasis coexist in endemic countries, affecting more than 1 billion people worldwide, and are recognised as major global vector-borne diseases. Tackling this disease requires an accurate diagnosis that is sensitive, specific, and rapid. This study aimed to describe and validate a new highly sensitive and specific multiple-analysis system that can effectively detect numerous etiological agents in a single test. Methods: A total of 230 human blood samples were assessed retrospectively for parasite characterisation, as well as 58 stool samples from non-human primates. Primers and probes were designed in the small subunit ribosomal RNA gene, except for Plasmodium spp., for which the novel target was Cytochrome Oxidase Subunit 1. Results: The analytical specificity of the presented method was 100%, with no unspecific amplifications or cross-reactions with other blood parasitic diseases. The detection limit obtained was between 0.6 and 3.01 parasites/µL for Plasmodium species, 1.8 parasites/mL for Trypanosomatidae, and 2 microfilariae/mL in the case of Filariae. The sensitivity, specificity, predictive values, and kappa coefficient reached almost 100%, except for Filariae, whose sensitivity dropped to 93.9% and whose negative predicted value dropped to 89.5%. The operational features described a turnaround and a hands-on time shorter than the compared methods with a lower cost per essay. Conclusions: This work presents a cost-effective and highly sensitive multiplexed tool (RT-PCR-bp) capable of performing simultaneous detection for blood parasitic diseases using specific fluorescence probes, enabling the diagnosis of low parasite loads and coinfections. Full article

Triatomines are vectors of Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae), the agent of Chagas disease. Stercorarian transmission occurs when infectious parasites are passed in insect feces to vertebrates through the biting wound or mucosa. Defecating on hosts during or shortly after blood feeding is, therefore, critical for transmission, and delayed triatomine defecation behavior has been posited to contribute to a low incidence of human Chagas disease in the U.S. We allowed nymphal T. cruzi-infected and uninfected Triatoma gerstaeckeri (Stål, 1859) and Triatoma sanguisuga (LeConte, 1855) to interact with restrained guinea pigs and measured insect feeding and defecation events; South American Rhodnius prolixus (Stål, 1859; Latin America) served as a comparison group. In 148 trials, 40.0% of insects fed, of which 71.2% defecated. Compared to R. prolixus, T. gerstaeckeri had >9 times higher odds of feeding, and T. sanguisuga fed longer. Observations of defecation while feeding occurred across all three species. The post-feeding defecation interval (PFDI) of R. prolixus was significantly shorter (4.54 ± 2.46 min) than that of T. gerstaeckeri (9.75 ± 2.52 min) and T. sanguisuga (20.69 ± 8.98 min). Furthermore, the PFDI was shorter for TcI-infected insects compared to uninfected insects. Triatoma gerstaeckeri and T. sanguisuga are capable of stercorarian transmission, although the calculated metrics suggest they are less efficient vectors than R. prolixus. Full article

The Trypanosomatidae family encompasses around 24 genera of unicellular protozoans, many of which are transmitted by various hematophagous arthropods, particularly members of the Orders Diptera and Hemiptera. Fleas and ticks—an understudied group of ectoparasites—have been shown to be hosts of a wide and crescent variety of trypanosomatid species. Further, fleas and ticks of companion animals have been particularly neglected in trypanosomatid surveillance despite the proximity to human populations and the anthropophagous habits of many of these arthropods, which can potentially act as vectors of zoonotic trypanosomatids. We aimed to identify the presence, characterize the species, and establish the prevalence of Kinetoplastids, including members of the Trypanosomatidae family, in ectoparasites collected from dogs and humans from Mexico. A total of 537 ectoparasite specimens belonging to six ectoparasite taxa (Amblyomma mixtum, A. tenellum, Ctenocephalides felis felis, Pulex simulans, Rhipicephalus linnaei, and Rh. sanguineus s.s.) were collected from 15 States of Mexico. An 800 bp fragment of the 18S-rDNA gene from kinetoplastids was amplified and sequenced. The presence of two agents (Trypanosoma caninum and Parabodo sp.) was detected in R. linnaei ticks and one (Blechomonas lauriereadi) in the cat flea Ct. felis felis. This is the first record of genetic material of kinetoplastid species in ectoparasites from dogs and humans in Mexico. Full article

Culex pipiens mosquitoes (Diptera: Culicidae) are widespread during warm periods and actively feed on blood while serving as vectors for various human and animal pathogens. Culex mosquitoes overwinter as adults in temperate zones, raising the question of whether hibernating Cx. pipiens can act as pathogen reservoirs. In this study, hibernating mosquitoes and mosquitoes collected during the warm season were tested for the presence of trypanosomatids and avian haemosporidian parasites using PCR. Midgut preparations were made from Cx. pipiens females in order to search for trypanosomatids morphologically. In total, 1037 Cx. pipiens mosquitoes (556 collected during the warm season and 481 overwintering mosquitos) were investigated. The parasite prevalence differed for mosquitoes collected during the warm season and hibernating ones for both Haemosporida (2.9% in warm-season and no infections in overwintering mosquitoes) and Trypanosomatida (1.6% and 0.4%, respectively) parasites. A phylogenetic analysis confirmed that the trypanosomatids found in hibernating mosquitoes were monoxenous and were not parasites of vertebrates. The peak prevalence of Haemosporida parasites was detected in July (4.9%) and August (2.8%), and for Trypanosomatida, it was detected in May (3.5%). The results of the present study show that overwintering Cx. pipiens mosquitoes are questionable reservoirs for avian haemosporidian parasites, but some monoxenous trypanosomatids can be found in overwintering females. Full article

Angomonas deanei belongs to Trypanosomatidae family, a family of parasites that only infect insects. It hosts a bacterial endosymbiont in a mutualistic relationship, constituting an excellent model for studying organelle origin and cellular evolution. A lipidomic approach, which allows for a comprehensive analysis of all lipids in a biological system (lipidome), is a useful tool for identifying and measuring different expression patterns of lipid classes. The present study applied GC-MS and NMR techniques, coupled with principal component analysis (PCA), in order to perform a comparative lipidomic study of wild and aposymbiotic A. deanei grown in the presence or absence of FBS. Unusual contents of branched-chain iso C17:0 and C19:0-cis-9,10 and-11,12 fatty acids were identified in A. deanei cultures, and it was interesting to note that their content slightly decreased at the log phase culture, indicating that in the latter growth stages the cell must promote the remodeling of lipid synthesis in order to maintain the fluidity of the membrane. The combination of analytical techniques used in this work allowed for the detection and characterization of lipids and relevant contributors in a variety of A. deanei growth conditions. Full article

The present work assessed the experimental susceptibility of Nyssomyia antunesi and Lutzomyia longipalpis to Leishmania (Viannia) lainsoni and L. (V.) lindenbergi. A L. (Leishmania) chagasi–Lu. longipalpis combination was used as a susceptible control. Wild-caught Ny. antunesi and laboratory-bred Lu. longipalpis were membrane-fed on blood with a 5 × 10⁶/mL log-phase promastigote culture suspension and dissected on days 2 and 8 post-blood meal (pbm) for analysis focused on the assessment of parasitoses, as well as placement and promastigote morphotyping. Survival curves were constructed. In all combinations, promastigotes were observed on day 8 pbm. For both Leishmania species, in Lu. longipalpis, the presence of parasites was observed up to the stomodeal valve, while in Ny. antunesi, the presence of parasites was observed up to the cardia. There were no significant differences in parasitosis between L. (V.) lainsoni and L. (V.) lindenbergi in either Ny. antunesi or Lu. longipalpis. Six morphological promastigote forms were distinguished in Giemsa-stained gut smears. The survival curves of all combinations decreased and were affected differently by several Lu. longipalpis–parasite combinations, as well with Lu. longipalpis–uninfected blood. These findings stress Lu. longipalpis as experimentally susceptible to Leishmania spp. and suggest the putative susceptibility of Ny. antunesi to L. (V.) lainsoni and L. (V.) lindenbergi. Full article

One of the major drawbacks of current treatments for neglected tropical diseases is the low safety of the drugs used and the emergence of resistance. Leishmaniasis is a group of neglected diseases caused by protozoa of the trypanosomatidae family that lacks preventive vaccines and whose pharmacological treatments are scarce and unsafe. Combination therapy is a strategy that could solve the above-mentioned problems, due to the participation of several mechanisms of action and the reduction in the amount of drug necessary to obtain the therapeutic effect. In addition, this approach also increases the odds of finding an effective drug following the repurposing strategy. From the previous screening of two collections of repositioning drugs, we found that pyrvinium pamoate had a potent leishmanicidal effect. For this reason, we decided to combine it separately with two clinically used leishmanicidal drugs, miltefosine and paromomycin. These combinations were tested in axenic amastigotes of Leishmania infantum obtained from bone marrow cells and in intramacrophagic amastigotes obtained from primary cultures of splenic cells, both cell types coming from experimentally infected mice. Some of the combinations showed synergistic behavior, especially in the case of the combination of pyrvinium pamoate with paromomycin, and exhibited low cytotoxicity and good tolerability on intestinal murine organoids, which reveal the potential of these combinations for the treatment of leishmaniasis. Full article

In the Old World, phlebotomine sand flies from the genus Phlebotomus are implicated in the transmission of Leishmania spp. parasites (Kinetoplastida: Trypanosomatidae) and viruses belonging to the genus Phlebovirus (Bunyavirales: Phenuiviridae). Two of the five sand fly species known to occur in Portugal, Phlebotomus perniciosus and Ph. ariasi, the former being the most ubiquitous, are recognized vectors of Leishmania infantum, which causes visceral leishmaniasis, the most prevalent form of leishmaniasis in the country. Phlebotomus perniciosus is also the vector of the neurotropic Toscana virus, which can cause aseptic meningitis. Entomological surveillance is essential to provide fundamental data about the presence of vectors and the pathogens they can carry. As such, and given the lack of data in Portugal, an entomological survey took place in the Algarve, the southernmost region of the country, from May to October 2018. Polymerase chain reaction assays were performed in order to detect the presence of the above-mentioned pathogens in sand fly pools. Not only were both Leishmania parasites and phleboviruses detected during this study, but more importantly, it was the first time their co-circulation was verified in the same sand fly population collected in Portugal. Full article

The Leishmaniinae subfamily of the Trypanosomatidae contains both genus Zelonia (monoxenous) and Endotrypanum (dixenous). They are amongst the nearest known relatives of Leishmania, which comprises many human pathogens widespread in the developing world. These closely related lineages are models for the genomic biology of monoxenous and dixenous parasites. Herein, we used comparative genomics to identify the orthologous groups (OGs) shared among 26 Leishmaniinae species to investigate gene family expansion/contraction and applied two phylogenomic approaches to confirm relationships within the subfamily. The Endotrypanum monterogeii and Zelonia costaricensis genomes were assembled, with sizes of 29.9 Mb and 38.0 Mb and 9.711 and 12.201 predicted protein-coding genes, respectively. The genome of E. monterogeii displayed a higher number of multicopy cell surface protein families, including glycoprotein 63 and glycoprotein 46, compared to Leishmania spp. The genome of Z. costaricensis presents expansions of BT1 and amino acid transporters and proteins containing leucine-rich repeat domains, as well as a loss of ABC-type transporters. In total, 415 and 85 lineage-specific OGs were identified in Z. costaricensis and E. monterogeii. The evolutionary relationships within the subfamily were confirmed using the supermatrix (3384 protein-coding genes) and supertree methods. Overall, this study showed new expansions of multigene families in monoxenous and dixenous parasites of the subfamily Leishmaniinae. Full article

Curcumin (CUR) is known for its wide folkloric effects on various infections; however, its solubility status has remained a hindrance to its bioavailability in the host. This study evaluated the comparative effects of CUR and CUR-nanoparticle in vitro on T. congolense, T. b. brucei, and T. evansi. Additionally, CUR and CUR-nanoparticle anti-Trypanosoma efficacy were assessed in vivo against T. congolense. All the CUR-nanoparticles were two folds more effective on the T. congolense as compared to CUR in vitro, with recorded efficacy of 3.67 ± 0.31; 7.61 ± 1.22; and 6.40 ± 3.07 μM, while the CUR-nanoparticles efficacy was 1.56 ± 0.50; 28.16 ± 9.43 and 13.12 ± 0.13 μM on T. congolense, T. b. brucei, and T. evansi, respectively. Both CUR and CUR-nanoparticles displayed moderate efficacy orally. The efficacy of CUR and CUR-nanoparticles in vivo was influenced by solubility, presence of food, and treatment period. CUR-treated mice were not cured of the infection; however, the survival rate of the orally treated mice was significantly prolonged as compared with intraperitoneal-treated mice. CUR-nanoparticles resulted in significant suppression of parasitemia even though relapsed was observed. In conclusion, CUR and CUR-nanoparticles possess moderate efficacy orally on the trypanosomes as compared to the intraperitoneal treatment. Full article

Trypanosomatids are mainly responsible for leishmaniasis, sleeping sickness, and Chagas disease, which are the most challenging among the neglected tropical diseases due to the problem of drug resistance. Although problems of target deconvolution and polypharmacology are encountered, a target-based approach is a rational method for screening drug candidates targeting a biomolecule that causes infections. The present study aims to summarize the latest information regarding potential inhibitors of squalene synthase and farnesyl phosphate synthase with anti-Trypanosomatidae activity. The information was obtained by referencing textbooks and major scientific databases from their inception until April 2023. Based on in vitro experiments, more than seventy compounds were reported to inhibit squalene synthase and farnesyl diphosphate synthase. Among these compounds, more than 30 were found to be active in vitro against Trypanosomatidae, inferring that these compounds can be used as scaffolds to develop new drugs against trypanosomatid-related infections. Overall, natural and synthetic products can inhibit enzymes that are crucial for the survival and virulence of trypanosomatids. Moreover, in vitro experiments have confirmed the activity of more than half of these inhibitors using cell-based assays. Nevertheless, additional studies on the cytotoxicity, pharmacokinetics, and lead optimization of potent anti-Trypanosomatid compounds should be investigated. Full article