Gene Editing in the Chagas Disease Vector Rhodnius prolixus by Cas9-Mediated ReMOT Control.

Rhodnius prolixus is currently the model vector of choice for studying Chagas disease transmission, a debilitating disease caused by Trypanosoma cruzi parasites. However, transgenesis and gene editing protocols to advance the field are still lacking. Here, we tested protocols for the maternal delivery of CRISPR-Cas9 (clustered regularly spaced palindromic repeats/Cas-9 associated) elements to developing R. prolixus oocytes and strategies for the identification of insertions and deletions (indels) in target loci of resulting gene-edited generation zero (G0) nymphs. We demonstrate successful gene editing of the eye color markers Rp-scarlet and Rp-white, and the cuticle color marker Rp-yellow, with highest effectiveness obtained using Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) with the ovary-targeting BtKV ligand. These results provide proof of concepts for generating somatic mutations in R. prolixus and potentially for generating germ line-edited lines in triatomines, laying the foundation for gene editing protocols that could lead to the development of novel control strategies for vectors of Chagas disease.

A near-complete genome assembly of Monochamus alternatus a major vector beetle of pinewood nematode.

The Japanese sawyer beetle, Monochamus alternatus, is not only one of the most important wood boring pest itself, but also a major vector of the invasive pinewood nematode (PWN), which is the causal agent of the devastative pine wilt disease (PWD) and threats the global pine forest. Here, we present a near-complete genome of M. alternatus at the chromosome level. The assembled genome was 792.05 Mb with contig N50 length of 55.99 Mb, which is the largest N50 size among the sequenced Coleoptera insects currently. 99.57% of sequence was anchored onto ten pseudochromosomes (one X-chromosome and nine autosomes), and the final genome harbored only 13 gaps. BUSCO evaluation revealed the presence of 99.0% of complete core genes. Thus, our genome assembly represented the highest-contiguity genome assembly as well as high completeness in insects so far. We identified 20,471 protein-coding genes, of which 20,070 (98.04%) were functionally annotated. The genome assembly of M. alternatus provides a valuable resource for exploring the evolution of the symbiosis between PWN and the vector insects.

Spatial distribution of Triatoma dimidiata peridomestic colonies modulated by distance between susceptible microhabitat patches.

Triatoma dimidiata is a vector of the hemoparasite Trypanosoma cruzi, the causal agent of Chagas disease. It settles reproductive colonies in the peridomicile of the premises. The peridomicile is comprised of a random set of artificial and natural features that overlap and assemble a network of microenvironmental suitable sites (patches) that interact with each other and favor the structure and proliferation of T. dimidiata colonies. The heterogeneity of patch characteristics hinders the understanding and identification of sites susceptible to colonization. In this study, a classification system using a random forest algorithm was used to identify peridomiciles susceptible to colonization to describe the spatial distribution of these sites and their relationship with the colonies of T. dimidiata in ten localities of Yucatan. From 1,000 peridomiciles reviewed, the classification showed that 13.9 % (139) of the patches were highly susceptible (HSP), and 86.1 % (861) were less susceptible (LSP). All localities had at least one HSP. The occupancy by patch type showed that the percentage of total occupancy and by colonies was higher in the HSP, while the occupancy by adult T. dimidiata without evidence of nymphs or exuviae (propagules) was higher in the LSP. A generalized additive model (GAM) revealed that the percentage of occupied patches increases as the abundance of individuals in the localities increases however, the percentage of occupied patches in LSP is lower than occupied in HSP. Distance analyses revealed that colonies and propagules were located significantly closer (approximately 200 m) to a colony in a HSP than any colony in a LSP. The distribution of T. dimidiata in the localities was defined by the distribution of patch type; as the occupancy in these patches increased, a network of peridomestic populations was configured, which may be promoted by a greater abundance of insects inside the localities. These results reveal that the spatial distribution of T. dimidiata individuals and colonies in the peridomicile at the locality scale corresponds to a metapopulation pattern within the localities through a system of patches mediated by distance and level of the vectors' occupancy.

The interplay between temperature, Trypanosoma cruzi parasite load, and nutrition: Their effects on the development and life-cycle of the Chagas disease vector Rhodnius prolixus.

Chagas disease, caused by the protozoan parasite Trypanosoma cruzi transmitted by blood-sucking insects of the subfamily Triatominae, is a major neglected tropical disease affecting 6 to 7 million of people worldwide. Rhodnius prolixus, one of the most important vectors of Chagas disease in Latin America, is known to be highly sensitive to environmental factors, including temperature. This study aimed to investigate the effects of different temperatures on R. prolixus development and life-cycle, its relationship with T. cruzi, and to gather information about the nutritional habits and energy consumption of R. prolixus. We exposed uninfected and infected R. prolixus to four different temperatures ranging from 24°C to 30°C, and monitored their survival, developmental rate, body and blood meal masses, urine production, and the temporal dynamics of parasite concentration in the excreted urine of the triatomines over the course of their development. Our results demonstrate that temperature significantly impacts R. prolixus development, life-cycle and their relationship with T. cruzi, as R. prolixus exposed to higher temperatures had a shorter developmental time and a higher mortality rate compared to those exposed to lower temperatures, as well as a lower ability to retain weight between blood meals. Infection also decreased the capacity of the triatomines to retain weight gained by blood-feeding to the next developmental stage, and this effect was proportional to parasite concentration in excreted urine. We also showed that T. cruzi multiplication varied depending on temperature, with the lowest temperature having the lowest parasite load. Our findings provide important insights into the potential impact of climate change on the epidemiology of Chagas disease, and can contribute to efforts to model the future distribution of this disease. Our study also raises new questions, highlighting the need for further research in order to understand the complex interactions between temperature, vector biology, and parasite transmission.

Diversity of Glossinidae (Diptera) species in The Gambia in relation to vegetation.

Glossina species are known to transmit African Trypanosomiasis, one of the most important infectious diseases for both livestock and humans in sub-Saharan Africa. Therefore, the aim of this study was to characterize trapped Glossina spp. from The Gambia using morphological and molecular techniques in relation to the vegetation cover types. A line transect survey was carried out in all the administrative regions of The Gambia. Tsetse fly trapping was carried out for 14 days during each season using line transect. A total of 220 Glossina spp. specimens (117 F and 103 M) were captured, and DNA was extracted from the legs of 100 randomly selected Glossina spp. Further, DNA samples were tested by a conventional PCR assay. A total of 135/220 (61%; 95% CI: 54.6-67.8%) and 85/220 (39%; 95% CI: 32.2-45.4%) flies were identified as Glossina morsitans submorsitans and Glossina palpalis gambiensis, respectively, with most caught during wet season (53.6%) and more females (53.2%) than males. Results of the morphological identification agreed with those of molecular identification. The type of vegetation cover significantly influenced the caught of tsetse flies. Animals and humans at the various trapping sites are at risk of being bitten by tsetse flies.

Epidemiology of visceral leishmaniasis in municipalities of Mato Grosso and the performance of surveillance activities: an updated investigation.

Visceral leishmaniasis (VL) is considered a globally neglected disease. To address the problem of VL endemic to Brazil, the Visceral Leishmaniasis Control Program (VLCP) was created, which recommends the development of health surveillance actions such as the identification of human and canine cases, vector control and prevention of disease. We aimed to investigate the epidemiological situation of VL in municipalities of the State of Mato Grosso (MT) and assess the execution of VLCP activities. Data on human cases were obtained from the Information System for Notifiable Diseases (SINAN), and data from entomological and canine inquiry were provided by the State's Health Department. Analyzes from the period 2019 - 2021 recorded 30 cases of human VL, distributed among 16 municipalities. Vectors were identified in 50% of the municipalities where entomological investigations were carried out, and the predominant specie was Lutzomyia longipalpis. A total of 15,585 dogs were subjected to serological examination, of which 18.91% tested seropositive for Leishmania infantum. However, it must be emphasized that only three municipalities conducted consecutive inquiries involving canine VL. Although VL is distributed widely throughout the State, only a few municipalities have undertaken the actions of the VLCP, thus highlighting the neglected status of the disease.

The perfect storm: Cutaneous leishmaniasis in troops deployed in the Atlantic forest of Argentina.

American cutaneous leishmaniasis (ACL) is the most prevalent form of leishmaniasis, associated with an ulcerative and stigmatizing mucocutaneous pathology. This study assessed the incidence of Leishmania (Viannia) braziliensis in members of the Argentine Army who were exposed to sandfly bites in Iguazú National Park (INP), northeastern Argentina, during an outbreak of ACL in 2019, and the presence of Leishmania in rodents, opossums and phlebotomine sandflies collected in the area of exposure. Samples from military personnel, wild animals and phlebotomine sandflies were analysed. A total of 20 (40%) patients among the Army personnel and two Akodon montensis rodents (11%) were positive for the presence of Leishmania sp. genes by PCR, while Nyssomyia whitmani and Migonemyia migonei, competent vectors of Leishmania, were also found at the same site. Sequences of hsp70 DNA fragments obtained from human samples confirmed the identity of L. (V.) braziliensis. The risk to which military personnel carrying out activities in the forest are exposed is highlighted, and this risk extends to any worker and visitor who circulates without protection in the INP, coming into contact with transmission "hot spots" due to the concentration of vectors, reservoirs and/or parasites.

Merging socioecological variables to predict risk of Chagas disease.

How far are we from predicting the occurrence of zoonotic diseases? In this paper we have made use of both socioecological and ecological variables to predict Chagas disease occurrence. Chagas disease involves, Trypanosoma cruzi, a complex life-cycle parasite which requires two hosts: blood-feeding triatomine insects and vertebrate hosts including humans. We have used a common risk assessment method combined with datasets that imply critical environmental and socioeconomic drivers of Chagas dynamics to predict the occurrence of this disease. We also carried out a network analysis to assess the interactions among triatomines and mammal host species given their human contact via whether hunted, domesticated or associated with anthropogenic landscapes in Mexico. We found that social backwardness variation, lack of health services and altitude had the largest relative influence Chagas events. Triatoma pallidipennis made use of the largest host diversity. Host species shared by the highest number of different triatomines were a woodrat, the highly appreciated bushmeat, and racoon. These results indicate both the predominance of socio-economic factors over ecological ones, and how close we are from predicting zoonotic diseases.

Trypanosoma cruzi infection reduces the population fitness of Mepraia spinolai, a Chagas disease vector.

The hematophagous insect Mepraia spinolai (Hemiptera: Reduviidae: Triatominae) is naturally infected with the protozoan parasite Trypanosoma cruzi, the agent of Chagas disease in humans. In this study, we compared the demographic parameters of M. spinolai with and without T. cruzi infection. We collected the immature life table data of 479 M. spinolai individuals of control cohort (reared on mice without T. cruzi infection) and 563 M. spinolai individuals of treatment cohort (reared on mice with T. cruzi infection). Nymphs were maintained in individual compartments inside a growth chamber (26°C; 65-75%) until adult emergence; moulting and survival were recorded daily. For the adult life table study of the control, we used 24 pairs of adults from the control cohort. For the adult life table study of T. cruzi-infected cohort, 25 infected females were paired with 25 males from the control cohort. Life table data were analysed using bootstrap-match technique based on the age-stage, two-sex life table. The preadult survival rate (0.5282) of the control cohort was significantly higher than that of the infected cohort (0.2913). However, the mean fecundity of reproductive females (Fr = 22.29 eggs/♀) and net reproductive rate of population (R0 = 5.07 offspring/individual) of the 0.5th percentile bootstrap-match control cohort were not significantly different from those of the infected cohort (Fr = 23.35 eggs/♀, R0 = 3.77 offspring/individual). Due to the shorter total preoviposition period and higher proportion of reproductive female, the intrinsic rate of increase (r = 0.0053 d-1 ) and finite rate of increase (λ = 1.0053 d-1 ) of control cohort of M. spinolai were significantly higher than those of the T. cruzi-infected cohort (r = 0.0035 d-1 , λ = 1.0035 d-1 ). These results suggest that T. cruzi infection reduces the population fitness of the Chagas disease vector M. spinolai.

Abundant triatomines in Texas dog kennel environments: Triatomine collections, infection with Trypanosoma cruzi, and blood feeding hosts.

Triatomine insects are vectors of the protozoan parasite Trypanosoma cruzi- the causative agent of Chagas disease. Chagas disease is endemic to Latin America and the southern United States and can cause severe cardiac damage in infected mammals, ranging from chronic disease to sudden death. Identifying interactions among triatomines, T. cruzi discrete typing units (DTUs), and blood feeding hosts is necessary to understand parasite transmission dynamics and effectively protect animal and human health. Through manual insect trapping efforts, kennel staff collections, and with the help of a trained scent detection dog, we collected triatomines from 10 multi-dog kennels across central and south Texas over a one-year period (2018-2019) and tested a subset to determine their T. cruzi infection status and identify the primary bloodmeal hosts. We collected 550 triatomines, including Triatoma gerstaeckeri (n = 515), Triatoma lecticularia (n = 15), Triatoma sanguisuga (n = 6), and Triatoma indictiva (n = 2), with an additional 10 nymphs and 2 adults unable to be identified to species. The trained dog collected 42 triatomines, including nymphs, from areas not previously considered vector habitat by the kennel owners. Using qPCR, we found a T. cruzi infection prevalence of 47 % (74/157), with T. lecticularia individuals more likely to be infected with T. cruzi than other species. Infected insects harbored two T. cruzi discrete typing units: TcI (64 %), TcIV (23 %), and mixed TcI/TcIV infections (13 %). Bloodmeal host identification was successful in 50/149 triatomines, revealing the majority (74 %) fed on a dog (Canis lupus), with other host species including humans (Homo sapiens), raccoons (Procyon lotor), chickens (Gallus gallus), wild pig (Sus scrofa), black vulture (Coragyps atratus), cat (Felis catus), and curve-billed thrasher (Toxostoma curviostre). Given the frequency of interactions between dogs and infected triatomines in these kennel environments, dogs may be an apt target for future vector control and T. cruzi intervention efforts.

Species diversity, barcode, detection of pathogens and blood meal pattern in Phlebotominae (Diptera: Psychodidae) from northeastern Mexico.

More than 90 species of phlebotomines are vectors of parasites, bacteria, and viruses, which cause disease in animals and humans. Therefore, their study is necessary to establish prevention and control strategies. Mexico is an endemic country for leishmaniasis, mostly in the center and southern regions of the country, yet only few studies have been conducted in the northern part of the country. The present study aims to: (a) assess the alpha diversity of Phlebotominae in an annual cycle, (b) to correlate climatic variables with abundance, (c) to generate barcodes of these insects as part of the integrative taxonomy, and (d) to detect Leishmania, Wolbachia and blood sources in an area close to where a case of autochthonous leishmaniasis has been detected in Nuevo Leon, Mexico. A systematic sampling was conducted during three consecutive nights from 17:00 to 22:00 h., placing Shannon traps, CDC traps with incandescent light, and BG Sentinel 2 + BG Lure traps. A total catch effort of 660 nights/traps/hours was achieved, in which a total number of 707 phlebotomines (58% female and 42% male) of six species were collected and identified. The most abundant species were Psathyromyia cratifer (57%) and Psathyromyia shannoni sensu stricto (26%). The highest abundance (72%; 507/707) was collected during March, April and May 2021. Barcodes were generated for four species of phlebotomines, which represent new records for Mexico. For the molecular detection of microorganisms, 302 specimens were analyzed, although no specimens were positive for Leishmania spp. Wolbachia strains were detected in phlebotomines with an infection rate of 1.32% (4/302) and found in Pa. cratifer and Lu. cruciata. Likewise, human DNA was identified in female Lu. cruciata and Pa. cratifer phlebotomines. These findings indicate the presence of potential vector species of the parasite Leishmania spp. This result shows the need for further entomological surveillance to elucidate the transmission mechanisms in these northern areas of the country.

Revised New World bioregions and environmental correlates for vectors of Chagas disease (Hemiptera, Triatominae).

The subfamily Triatominae includes a group of hematophagous insects, vectors of the parasite Trypanosoma cruzi, which is the etiological agent of Chagas disease, also known as American trypanosomiasis. Triatomines occur in the Old and New World and occupy diverse habitats including tropical and temperate areas. Some studies suggest the distributions of triatomines group into three or four regions. This study objectively determined bioregions focused specifically on New World Triatominae, using clustering and ordination analysis. We also identified indicator species by bioregion and investigated relationships among bioregions and environmental variables using redundancy analysis and multivariate regression trees. We delineated seven bioregions specific to Triatominae and linked each with indicator species. This result suggests more biogeographical structure exists than was revealed in earlier studies that were more general, subjective, and based on older taxonomic and distributional information. Precipitation, elevation, and vegetation were important variables in the delimitating bioregions. This implies that more detailed study of how these factors influence triatomine distributions could benefit understanding of how Chagas disease is spread.

Spatiotemporal changes in exposition risk to leishmaniases vector in residences within a fishing tourism area of Pantanal wetland.

Miranda Municipality of Mato Grosso do Sul, borders the Pantanal wetland, a famous fishing destination visited by tourists from all over the world, and is a location where visceral leishmaniasis has been reported. To assess the risk of Leishmania infantum transmission, we studied the sandfly community, focusing on known vector and parasite presence. We conducted light trap collections twice per month at nine sites within the city (including two forested areas) for one year. We collected a total of 12,727 sand flies, 10,891 males and 1,836 females belonging to 11 species: Brumptomyia avellari, Evandromyia aldafalcaoae, Ev. evandroi, Ev. lenti, Ev. sallesi, Ev. walkeri, Lu. longipalpis, Nyssomyia whitmani, Psathyromyia bigeniculata, Pa. hermanlenti and Pa. punctigeniculata. Lutzomyia longipalpis, the proven vector of Leishmania infantum, was captured each month, and was the most abundant species observed, accounting for more than 99% of sand flies captured in most sites, especially where chicken coops were present. Evidence of Leishmania infantum infection was detected in 0.40% of Lu. longipalpis tested. We developed a generalized mixed multilevel model for Lu. longipalpis, that includes within-year seasonality, location of capture (indoors vs. outdoors), vector abundance, and sex ratio. The VL vector was abundant both inside and outside houses. Large numbers of Lu. longipalpis were observed in outdoor sites where domestic animals were present but were absent from forest sites. Our findings suggest high vector populations and Le. infantum presence in a city where tourists could be exposed to visceral leishmaniasis, with significant implications for more surveillance and control activities.

Changes in contributions of different Anopheles vector species to malaria transmission in east and southern Africa from 2000 to 2022.

BACKGROUND: Malaria transmission in Africa is facilitated by multiple species of Anopheles mosquitoes. These vectors have different behaviors and vectorial capacities and are affected differently by vector control interventions, such as insecticide-treated nets and indoor residual spraying. This review aimed to assess changes in the contribution of different vector species to malaria transmission in east and southern Africa over 20 years of widespread insecticide-based vector control. METHODS: We searched PubMed, Global Health, and Web of Science online databases for articles published between January 2000 and April 2023 that provided species-specific sporozoite rates for different malaria vectors in east and southern Africa. We extracted data on study characteristics, biting rates, sporozoite infection proportions, and entomological inoculation rates (EIR). Using EIR data, the proportional contribution of each species to malaria transmission was estimated. RESULTS: Studies conducted between 2000 and 2010 identified the Anopheles gambiae complex as the primary malaria vector, while studies conducted from 2011 to 2021 indicated the dominance of Anopheles funestus. From 2000 to 2010, in 57% of sites, An. gambiae demonstrated higher parasite infection prevalence than other Anopheles species. Anopheles gambiae also accounted for over 50% of EIR in 76% of the study sites. Conversely, from 2011 to 2021, An. funestus dominated with higher infection rates than other Anopheles in 58% of sites and a majority EIR contribution in 63% of sites. This trend coincided with a decline in overall EIR and the proportion of sporozoite-infected An. gambiae. The main vectors in the An. gambiae complex in the region were Anopheles arabiensis and An. gambiae sensu stricto (s.s.), while the important member of the An. funestus group was An. funestus s.s. CONCLUSION: The contribution of different vector species in malaria transmission has changed over the past 20 years. As the role of An. gambiae has declined, An. funestus now appears to be dominant in most settings in east and southern Africa. Other secondary vector species may play minor roles in specific localities. To improve malaria control in the region, vector control should be optimized to match these entomological trends, considering the different ecologies and behaviors of the dominant vector species.

Acute Chagas Disease Presenting as Preseptal Cellulitis.

Chagas disease, also known as American trypanosomiasis, is caused by Trypanosoma cruzi, a parasite transmitted by hematophagous triatomine insects (subfamily Triatominae) belonging to the Reduviidae family, order Hemiptera. Infection occurs through contact with the feces of the infected vector at the site of its bite or on intact mucosa. [Pediatr Ann. 2023;52(10):e394-e397.].

Morphological and molecular evidence of sandfly (Diptera: Psychodidae: Phlebotomine) and its relevance to recent cases of leishmaniasis from Jammu region of North India.

Present study was conducted to carry out morphological and molecular confirmation of sandflies collected at the Department of Parasitology, Faculty of Veterinary and Animal Sciences, R.S. Pura, Jammu, India. Larva was maggot like with large head, thorax and abdomen with typical black head, 12 abdominal segments and last abdominal segment carried two pairs of caudal bristles with matchstick hairs on each segment. The adult fly possessed head, abdomen and thorax. Head consisted of pair of long, hairy and beaded antenna, proboscis and one pair of prominent black eyes. Thorax possessed a pair of wings and three pairs of legs, wings were hairy and pointed with 2nd longitudinal vein branched twice. The abdominal segments were covered with small hairs and last abdominal segment was having a pair of anal recti. These identification characteristics confirmed the fly under study as Phlebotomus argentipes that confirms its occurance in this region. Molecular characterization of identified flies was carried out on positive morophological flies. Confirmation of Phlebotomus species was ascertained by amplifying the 18S ribosomal RNA gene sequence using PCR. Clear amplification was observed for Phlebotomus argentipes (538 bp). After sequencing/genotyping, Phlebotomus argentipes (OP646634) isolate of present study was clustering in same clade with Phlebotomus argentipes sequences obtained from GeneBank from other locations across globe, irrespective of their geographical location, thus providing the molecular evidence of this species present in Jammu region of North India.

Canine Systemic Insecticides Fluralaner and Lotilaner Induce Acute Mortality of Triatoma gerstaeckeri, North American Vector of the Chagas Disease Parasite.

Chagas disease is a health concern for humans and animals across the Americas, and control options targeting the triatomine vectors of Trypanosoma cruzi, the causative agent of Chagas disease, are limited. Host-targeted interventions may be a useful and underused tool in controlling the spread of T. cruzi from vectors to hosts. Domestic dogs are known to be key bloodmeal hosts for triatomines as well as T. cruzi reservoirs and may be an effective and practical target for host-targeted insecticide deployment. We hypothesized that treating dogs with commercially available systemic insecticides (labeled for flea and tick control) would result in mortality of triatomines after consuming treated blood. We enrolled 16 privately owned dogs into five treatment groups to receive either fluralaner (Bravecto) or lotilaner (Credelio), alone or in combination with ivermectin. Blood from dogs before the initiation of treatment served as controls. Blood was collected 0, 7, 30, 45, and 90 days after the initial canine insecticide treatment and fed to 10 Triatoma gerstaeckeri nymphs through a membrane feeder, and survival was tracked daily for 7 days and weekly thereafter. All triatomines in the control and ivermectin groups survived the initial period, with no significant difference in long-term survival. In contrast, 99.7% of triatomines that fed on blood from dogs treated with either fluralaner or lotilaner died within 3 days. Although the impact of canine treatment on suppressing vector populations is unknown, fluralaner and lotilaner appear to be a compelling option for an integrated vector management approach to triatomine control.

Biting rhythms and infection rates of anthropophilic sand fly species (Diptera: Phlebotominae) in sites with different land use in southern Mexico.

Could tropical forest conversion shape sand fly (Diptera: Phlebotominae) biting rhythms and Leishmania infection rates? Using a Shannon trap, we estimated the bite rate and infection prevalence among anthropophilic sand flies at sites with different land use in southern Mexico. We estimated the expected monthly infection rate of the Leishmania parasite along the gradient and generated information on the biting rhythm of sand flies in a poorly characterized cutaneous leishmaniasis endemic region. We used generalized mixed linear and mixed additives models to evaluate differences in the biting rate, nocturnal activity, and inoculation rate of female sand flies, as well as their relationship with the loss of forest cover and environmental disparities recorded throughout the study area. Our results show that the loss of forest cover influences the biting rhythm of sand fly species and the potential number of infectious bites with Leishmania, but the greatest entomological and potential epidemiological risk continues to be associated with sylvatic areas (amplification events). Despite this, we detected that the effect of forest cover (%) on the entomological exposure seems to be also dependent on the sand fly species, and that, albeit to a lesser extent, Leishmania parasite is circulating in disturbed landscapes through generalist and competent sand fly vector species. We also found that land use change did not affect the nocturnal activity, however we detected that important vector species were active most of the time. Contrary to our expectation, temperature and humidity did not shape the biting rhythm of sand fly species. We discuss the limitations and epidemiological implications of our findings regarding the risk of contracting leishmaniasis in southern Mexico.

Insights into the microRNA landscape of Rhodnius prolixus, a vector of Chagas disease.

The growing interest in microRNAs (miRNAs) over recent years has led to their characterization in numerous organisms. However, there is currently a lack of data available on miRNAs from triatomine bugs (Reduviidae: Triatominae), which are the vectors of the protozoan parasite Trypanosoma cruzi, the causative agent of Chagas disease. A comprehensive understanding of the molecular biology of vectors provides new insights into insect-host interactions and insect control approaches, which are key methods to prevent disease incidence in endemic areas. In this work, we describe the miRNome profiles from gut, hemolymph, and salivary gland tissues of the Rhodnius prolixus triatomine. Small RNA sequencing data revealed abundant expression of miRNAs, along with tRNA- and rRNA-derived fragments. Fifty-two mature miRNAs, previously reported in Ecdysozoa, were identified, including 39 ubiquitously expressed in the three tissues. Additionally, 112, 73, and 78 novel miRNAs were predicted in the gut, hemolymph, and salivary glands, respectively. In silico prediction showed that the top eight most highly expressed miRNAs from salivary glands potentially target human blood-expressed genes, suggesting that R. prolixus may modulate the host's gene expression at the bite site. This study provides the first characterization of miRNAs in a Triatominae species, shedding light on the role of these crucial regulatory molecules.

The Pampa del Indio project: sustainable vector control and long-term declines in the prevalence and abundance of Triatoma infestans infected with Trypanosoma cruzi in the Argentine Chaco.

BACKGROUND: The Gran Chaco region is a major hotspot of Chagas disease. We implemented a 9-year program aimed at suppressing house infestation with Triatoma infestans and stopping vector-borne transmission to creole and indigenous (Qom) residents across Pampa del Indio municipality (Argentine Chaco). The aim of the present study was to assess the intervention effects on parasite-based transmission indices and the spatial distribution of the parasite, and test whether house-level variations in triatomine infection with Trypanosoma cruzi declined postintervention and were influenced by household ethnicity, persistent infestation linked to pyrethroid resistance and other determinants of bug infection. METHODS: This longitudinal study assessed house infestation and bug infection with T. cruzi before and after spraying houses with pyrethroids and implemented systematic surveillance-and-response measures across four operational areas over the period 2007-2016. Live triatomines were individually examined for infection by optical microscopy or kinetoplast DNA (kDNA)-PCR and declared to be infected with T. cruzi when assessed positive by either method. RESULTS: The prevalence of infection with T. cruzi was 19.4% among 6397 T. infestans examined. Infection ranged widely among the study areas (12.5-26.0%), household ethnicity (15.3-26.9%), bug ecotopes (1.8-27.2%) and developmental stages (5.9-27.6%), and decreased from 24.1% (baseline) to 0.9% (endpoint). Using random-intercept multiple logistic regression, the relative odds of bug infection strongly decreased as the intervention period progressed, and increased with baseline domestic infestation and bug stage and in Qom households. The abundance of infected bugs and the proportion of houses with ≥ 1 infected bug remained depressed postintervention and were more informative of area-wide risk status than the prevalence of bug infection. Global spatial analysis revealed sharp changes in the aggregation of bug infection after the attack phase. Baseline domestic infestation and baseline bug infection strongly predicted the future occurrence of bug infection, as did persistent domestic infestation in the area with multiple pyrethroid-resistant foci. Only 19% of houses had a baseline domestic infestation and 56% had ever had ≥ 1 infected bug. CONCLUSIONS: Persistent bug infection postintervention was closely associated with persistent foci generated by pyrethroid resistance. Postintervention parasite-based indices closely agreed with human serosurveys at the study endpoint, suggesting transmission blockage. The program identified households and population subgroups for targeted interventions and opened new opportunities for risk prioritization and sustainable vector control and disease prevention.