Optimising the surveillance of Aedes aegypti in Brazil by selecting smaller representative areas within an endemic city.

OBJECTIVES: Arboviruses, such as dengue (DENV), zika (ZIKV), and chikungunya (CHIKV), constitute a growing urban public health threat. Focusing on Aedes aegypti mosquitoes, their primary vectors, is crucial for mitigation. While traditional immature-stage mosquito surveillance has limitations, capturing adult mosquitoes through traps yields more accurate data on disease transmission. However, deploying traps presents logistical and financial challenges, demonstrating effective temporal predictions but lacking spatial accuracy. Our goal is to identify smaller representative areas within cities to enhance the early warning system for DENV outbreaks. METHODS: We created Sentinel Geographic Units (SGUs), smaller areas of 1 km2 within each stratum, larger areas, with the aim of aligning the Trap Positivity Index (TPI) and Adult Density Index (ADI) with their respective strata. We conducted a two-step evaluation of SGUs. First, we examined the equivalence of TPI and ADI between SGUs and strata from January 2017 to July 2022. Second, we assessed the ability of SGU's TPI and ADI to predict DENV outbreaks in comparison to Foz do Iguaçu's Early-Warning System, which forecasts outbreaks up to 4 weeks ahead. Spatial and temporal analyses were carried out, including data interpolation and model selection based on Akaike information criteria (AIC). RESULTS: Entomological indicators produced in small SGUs can effectively replace larger sentinel areas to access dengue outbreaks. Based on historical data, the best predictive capability is achieved 2 weeks after infestation verification. Implementing the SGU strategy with more frequent sampling can provide more precise space-time estimates and enhance dengue control. CONCLUSIONS: The implementation of SGUs offers an efficient way to monitor mosquito populations, reducing the need for extensive resources. This approach has the potential to improve dengue transmission management and enhance the public health response in endemic cities.

Citywide Integrated Aedes aegypti Mosquito Surveillance as Early Warning System for Arbovirus Transmission, Brazil.

Arbovirus epidemiology lacks efficient and timely surveillance systems with accurate outbreak alert signals. We devised a citywide integrated surveillance system combining entomologic, epidemiologic, and entomo-virologic data gathered during 2017-2020 in Foz do Iguaçu, Brazil. We installed 3,476 adult mosquito traps across the city and inspected traps every 2 months. We compared 5 entomologic indices: traditional house and Breteau indices for larval surveys and trap positivity, adult density, and mosquitoes per inhabitant indices for adult trapping. We screened for dengue, Zika, and chikungunya viruses in live adult Aedes aegypti mosquitoes collected from traps. Indices based on adult mosquito sampling had higher outbreak predictive values than larval indices, and we were able to build choropleth maps of infestation levels <36 h after each round of trap inspection. Locating naturally infected vectors provides a timely support tool for local public health managers to prioritize areas for intervention response to prevent virus outbreaks.

Infection in asymptomatic carriers of SARS-CoV-2 can interfere with the achievement of robust immunity on a population scale.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread worldwide as a severe pandemic, and a significant portion of the infected population may remain asymptomatic. Given this, five surveys were carried out between May and September 2020 with a total of 3585 volunteers in the municipality of Foz do Iguaçu, State of Paraná, a triple border region between Brazil/Argentina/Paraguay. Five months after the first infection, volunteers were re-analysed for the production of IgG anti-Spike and anti-RBD-Spike, in addition to analyses of cellular immunity. Seroconversion rates ranged from 4.4 % to a peak of 37.21 % followed by a reduction in seroconversion to 21.1 % in September, indicating that 25 % of the population lost their circulating anti-SARS-CoV-2 antibodies 3 months after infection. Analyses after 5 months of infection showed that only 17.2 % of people still had anti-RBD-Spike antibodies, however, most volunteers had some degree of cellular immune response. The strategy of letting people become naturally infected with SARS-CoV-2 to achieve herd immunity is flawed, and the first contact with the virus may not generate enough immunogenic stimulus to prevent a possible second infection.

Malathion insecticide resistance in Aedes aegypti: laboratory conditions and in situ experimental approach through adult entomological surveillance.

OBJECTIVE: In Brazil, the most common method of controlling outbreaks of arbovirus is by the use of chemical sprays, which kill the insect vector, Aedes aegypti. The main objective of this study was to evaluate the resistance of Ae. aegypti to the insecticide, malathion, in situ. The location of this study was the municipality of Foz do Iguaçu, in the state of Paraná, Brazil. METHODS: Ultra-low-volume (ULV) fogging equipment was used, by vehicle, to apply the insecticide in situ, and mosquito populations after treatment were compared with those of control areas. The resistance of strains collected from the municipality was compared to the Rockefeller strain under laboratory conditions. RESULTS: We found 220 adult female specimens and 7423 eggs of Ae. aegypti in the areas subjected to UBV treatment, whereas 245 adult females and 10 557 eggs were found in the control areas. The UBV treatment area showed no significant difference compared to the control area, for all the indices. Mortality of the Rockefeller colony varied more quickly when there were slight variations in malathion concentration than the Foz do Iguaçu population.

OBJECTIF: Au Brésil, la méthode la plus courante de lutte contre les épidémies d'arbovirus consiste à utiliser des pulvérisations chimiques qui tuent l'insecte vecteur, Aedes aegypti. L'objectif principal de cette étude était d'évaluer la résistance de Ae. aegypti à l'insecticide, le malathion, in situ. Le lieu de cette étude était la municipalité de Foz do Iguaçu, dans l'état du Paraná, au Brésil. MÉTHODES: Un équipement de brumisation à très faible volume (ULV) a été utilisé, par véhicule, pour appliquer l'insecticide in situ et les populations de moustiques après le traitement ont été comparées à celles des zones témoins. La résistance des souches collectées dans la municipalité a été comparée à la souche Rockefeller dans des conditions de laboratoire. RÉSULTATS: Nous avons trouvé 220 spécimens femelles adultes et 7.423 œufs d'Ae. aegypti dans les zones soumises au traitement ULV, alors que 245 femelles adultes et 10.557 œufs ont été trouvés dans les zones témoins. La zone de traitement ULV n'a montré aucune différence significative par rapport à la zone témoin, pour tous les indices. La mortalité de la colonie de Rockefeller variait plus rapidement lorsqu'il y avait de légères variations dans la concentration de malathion que la population de Foz do Iguaçu.

Automated classification of mixed populations of Aedes aegypti and Culex quinquefasciatus mosquitoes under field conditions.

BACKGROUND: The recent rise in the transmission of mosquito-borne diseases such as dengue virus (DENV), Zika (ZIKV), chikungunya (CHIKV), Oropouche (OROV), and West Nile (WNV) is a major concern for public health managers worldwide. Emerging technologies for automated remote mosquito classification can be supplemented to improve surveillance systems and provide valuable information regarding mosquito vector catches in real time. METHODS: We coupled an optical sensor to the entrance of a standard mosquito suction trap (BG-Mosquitaire) to record 9151 insect flights in two Brazilian cities: Rio de Janeiro and Brasilia. The traps and sensors remained in the field for approximately 1 year. A total of 1383 mosquito flights were recorded from the target species: Aedes aegypti and Culex quinquefasciatus. Mosquito classification was based on previous models developed and trained using European populations of Aedes albopictus and Culex pipiens. RESULTS: The VECTRACK sensor was able to discriminate the target mosquitoes (Aedes and Culex genera) from non-target insects with an accuracy of 99.8%. Considering only mosquito vectors, the classification between Aedes and Culex achieved an accuracy of 93.7%. The sex classification worked better for Cx. quinquefasciatus (accuracy: 95%; specificity: 95.3%) than for Ae. aegypti (accuracy: 92.1%; specificity: 88.4%). CONCLUSIONS: The data reported herein show high accuracy, sensitivity, specificity and precision of an automated optical sensor in classifying target mosquito species, genus and sex. Similar results were obtained in two different Brazilian cities, suggesting high reliability of our findings. Surprisingly, the model developed for European populations of Ae. albopictus worked well for Brazilian Ae. aegypti populations, and the model developed and trained for Cx. pipiens was able to classify Brazilian Cx. quinquefasciatus populations. Our findings suggest this optical sensor can be integrated into mosquito surveillance methods and generate accurate automatic real-time monitoring of medically relevant mosquito species.

Spatial analysis of dengue transmission in an endemic city in Brazil reveals high spatial structuring on local dengue transmission dynamics.

In the last decades, dengue has become one of the most widespread mosquito-borne arboviruses in the world, with an increasing incidence in tropical and temperate regions. The mosquito Aedes aegypti is the dengue primary vector and is more abundant in highly urbanized areas. Traditional vector control methods have showing limited efficacy in sustaining mosquito population at low levels to prevent dengue virus outbreaks. Considering disease transmission is not evenly distributed in the territory, one perspective to enhance vector control efficacy relies on identifying the areas that concentrate arbovirus transmission within an endemic city, i.e., the hotspots. Herein, we used a 13-month timescale during the SARS-Cov-2 pandemic and its forced reduction in human mobility and social isolation to investigate the spatiotemporal association between dengue transmission in children and entomological indexes based on adult Ae. aegypti trapping. Dengue cases and the indexes Trap Positive Index (TPI) and Adult Density Index (ADI) varied seasonally, as expected: more than 51% of cases were notified on the first 2 months of the study, and higher infestation was observed in warmer months. The Moran's Eigenvector Maps (MEM) and Generalized Linear Models (GLM) revealed a strong large-scale spatial structuring in the positive dengue cases, with an unexpected negative correlation between dengue transmission and ADI. Overall, the global model and the purely spatial model presented a better fit to data. Our results show high spatial structure and low correlation between entomological and epidemiological data in Foz do Iguaçu dengue transmission dynamics, suggesting the role of human mobility might be overestimated and that other factors not evaluated herein could be playing a significant role in governing dengue transmission.

Entomo-virological surveillance followed by serological active survey of symptomatic individuals is helpful to identify hotspots of early arbovirus transmission.

Arboviruses transmitted by Aedes aegypti in urban environments have spread rapidly worldwide, causing great impacts on public health. The development of reliable and timely alert signals is among the most important steps in designing accurate surveillance systems for vector-borne diseases. In July and September 2017, we conducted a pilot study to improve an existing integrated surveillance system by using entomo-virological surveillance to prioritize areas to conduct active searches for individuals with arbovirus infection symptoms. Foz do Iguaçu City has a permanent entomo-virological surveillance system with approximately 3,500 traps to capture Aedes sp. in the adult stage. The Aedes aegypti females are captured alive and human samples are submitted to RT-qPCR (real-time qPCR) screening for DENV, ZIKV, and CHIKV diagnosis. Of the 55 Ae. aegypti mosquitoes tested in July 2017, seven (12.7%) were considered positive for DENV-2 and three (5.4%) for CHIKV. In September, we tested a sample of 54 mosquitoes, and 15 (27.7%) were considered infected by DENV-2. We created 25 circumferences with 150-m radius each to perform an active survey to identify symptomatic householders. In July, we selected one circumference, and five (35.7%) patients were positive for DENV, whereas two (14.3%) for CHIKV. In September, we selected four circumferences, and, from the 21 individuals sampled, nine (42.8%) were positive for DENV-2. A statistical model with a binomial response was used to estimate the number of cases in areas without active surveys, i.e., 20 circumferences. We estimated an additional 83 symptomatic patients (95% CI: 45-145) to be found in active searches, with 38 (95% CI: 18-72) of them confirming arbovirus infection. Arbovirus detection and serotyping in mosquitoes, but also in symptomatic individuals during active surveys, can provide an alert signal of early arbovirus transmission.

Entomo-Virological Aedes aegypti Surveillance Applied for Prediction of Dengue Transmission: A Spatio-Temporal Modeling Study.

Currently, DENV transmitted primarily by Aedes aegypti affects approximately one in three people annually. The spatio-temporal heterogeneity of vector infestation and the intensity of arbovirus transmission require surveillance capable of predicting an outbreak. In this work, we used data from 4 years of reported dengue cases and entomological indicators of adult Aedes collected from approximately 3500 traps installed in the city of Foz do Iguaçu, Brazil, to evaluate the spatial and temporal association between vector infestation and the occurrence of dengue cases. Entomological (TPI, ADI and MII) and entomo-virological (EVI) indexes were generated with the goal to provide local health managers with a transmission risk stratification that allows targeting areas for vector control activities. We observed a dynamic pattern in the evaluation; however, it was a low spatio-temporal correlation of Ae. aegypti and incidence of dengue. Independent temporal and spatial effects capture a significant portion of the signal given by human arbovirus cases. The entomo-virological index (EVI) significantly signaled risk in a few areas, whereas entomological indexes were not effective in providing dengue risk alert. Investigating the variation of biotic and abiotic factors between areas with and without correlation should provide more information about the local epidemiology of dengue.

One health approach on serosurvey of anti-Leptospira spp. in homeless persons and their dogs in South Brazil.

Although leptospirosis has been described as a worldwide bacterial zoonosis primarily affecting vulnerable populations, to date no study has focused on concomitant serosurvey of homeless persons and their dogs. The aim of the present study was, to use a One Health approach to serologically assess homeless persons and their dogs in 3 major cities of south Brazil (São Paulo, Curitiba, and Foz do Iguaçu). Environmental information was obtained with an epidemiological questionnaire given to all participants. A total of 200 human and 75 dog samples were tested for anti-Leptospira spp. antibodies to thirty different serovars using the microscopic agglutination test. None of the homeless persons were positive while 5 of the 75 (6.7%) dogs were positive. Among homeless population, 89% (177 of 200) were male, 61% (122 of 200) self-declared Non-white, and 67% (134 of 200) were educated up to the 8th school grade. Lower exposure of homeless persons to Leptospira spp. in the present study when compared to other vulnerable populations (slum and low-income residents) may be result of less direct exposure as they are able to rapidly change locations in response to flooding events. In addition, these results may reflect the effectiveness of a specific healthcare service provided to people living in the streets in the 3 cities. While dogs may be used as environmental sentinels for leptospirosis, the low seropositivity results found in this report may indicate low transmission risk to homeless owners in direct daily contact with their dogs.

Adaptation of a Human Diagnostic Kit to Detect Dengue, Zika, and Chikungunya Viruses in Mosquito Samples (Aedes aegypti and Aedes albopictus): A Contribution to Public Health in the International Triple Border (Brazil, Paraguay, and Argentina).

Objective: The objective of this work was to adapt a diagnostic kit developed for humans to identify Dengue (DENV1, DENV2, DENV3, DENV4), Zika (ZIKV) and Chikungunya virus (CHIKV) in females of Aedes aegypti and Aedes albopictus and to verify if the occurrence of mosquitoes infected with these three arboviruses are being found in regions with high occurrence of these diseases in humans. Materials and Methods: For this purpose, live mosquitoes were captured between January and June 2020 using 3,476 traps permanently installed in the field were used. After capture, the species were identified, then the females were placed in a pool of 2 to 10 specimens and sent to the laboratory for detection of DENV1, DENV2, DENV3, DENV4, ZIKV and CHIKV by RT-PCR using a commercial human kit for arboviruses. Results: Of the 76 mosquito pools collected, six (7.9%) pools tested positive for the DENV2 virus. The DENV-positive mosquitoes were collected in regions with a high incidence of reported cases of Dengue or in adjacent areas. Conclusion: The absence of kits for the detection of these arboviruses in Aedes is a limiting factor and the adequacy of commercial kits, already used for the diagnosis of arboviruses in humans, the results presented demonstrate that it is possible to identify the presence of DENV2 in mosquitoes with the respective kit, reinforcing the use of RT-qPCR as a robust diagnostic tool for epidemiological surveillance allowing managers to receive timely results for decision-making regarding prevention and control actions.

The adoption of the One Health approach to improve surveillance of venomous animal injury, vector-borne and zoonotic diseases in Foz do Iguaçu, Brazil.

Public health institutions with sectorized structure and low integration among field teams, old-fashioned practices such as paper-based storage system, and poorly qualified health agents have limited ability to conduct accurate surveillance and design effective timely interventions. Herein, we describe the steps taken by the Zoonosis Control Center of Foz do Iguaçu (CCZ-Foz) in the last 23 years to move from an archaic and sectorized structure to a modern and timely surveillance program embracing zoonotic diseases, venomous animal injuries, and vector-borne diseases epidemiology under the One Health approach. The full implementation of the One Health approach was based on 5 axes: (1) merging sectorized field teams; (2) adoption of digital solutions; (3) health agents empowerment and permanent capacitation; (4) social mobilization; and (5) active surveys. By doing so, notifications related to zoonotic diseases and venomous animals increased 10 and 21 times, respectively, with no impairment on arbovirus surveillance (major concern in the city). Open sources database (PostgreSQL) and software (QGis) are daily updated and create real-time maps to support timely decisions. The adoption of One Health approach increased preparedness for endemic diseases and reemerging and emerging threats such as Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2).

'Candidatus Mycoplasma haemoalbiventris' and tick-borne pathogens screening in white-eared opossums (Didelphis albiventris) from Curitiba and Foz do Iguaçu Cities, Paraná State, southern Brazil.

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.

Mosquitoes infected with dengue viruses in Brazil.

Dengue epidemics have been reported in Brazil since 1985. The scenery has worsened in the last decade because several serotypes are circulating and producing a hyper-endemic situation, with an increase of DHF/DSS cases as well as the number of fatalities. Herein, we report dengue virus surveillance in mosquitoes using a Flavivirus genus-specific RT-Hemi-Nested-PCR assay. The mosquitoes (Culicidae, n = 1700) collected in the Northeast, Southeast and South of Brazil, between 1999 and 2005, were grouped into 154 pools. Putative genomes of DENV-1, -2 and -3 were detected in 6 mosquito pools (3.8%). One amplicon of putative DENV-1 was detected in a pool of Haemagogus leucocelaenus suggesting that this virus could be involved in a sylvatic cycle. DENV-3 was found infecting 3 pools of larvae of Aedes albopictus and the nucleotide sequence of one of these viruses was identified as DENV-3 of genotype III, phylogenetically related to other DENV-3 isolated in Brazil. This is the first report of a nucleotide sequence of DENV-3 from larvae of Aedes albopictus.

Molecular Detection of Dengue Virus in Mosquitoes as an Early Indicator to Aid in the Prevention of Human Infection in Endemic Areas.

Human cases of dengue virus based on the National Dengue Control Plan were compared with the molecular detection of the dengue virus in trapped mosquitoes, verifying the prediction and efficacy potentials of vector control between the two methodologies in a city with three endemic frontiers. Molecular detection of dengue virus in trapped mosquitoes was significantly higher than in human cases (p = 0.0435). Thus, molecular detection could be used as an early indicator to help prevent more human cases of dengue.

Exploring deeper genetic structures: Aedes aegypti in Brazil.

Aedes aegypti, being the principal vector of dengue (DENV1 to 4), chikungunya and Zika viruses, is considered as one of the most important mosquito vectors. In Brazil, despite regular vector control programs, Ae. aegypti still persists with high urban density in all the states. This study aimed to estimate the intra and inter population genetic diversity and genetic structure among 15 Brazilian populations of Ae. aegypti based on 12 microsatellite loci. A total of 510 specimens were analyzed comprising eight locations from northern (Itacoatiara, Manaus, Novo Airão, Boa Vista, Rio Branco, Porto Velho, Guajará-Mirim and Macapá), three from southeastern (Araçatuba, São José de Rio Preto and Taubaté), one from southern (Foz do Iguaçu), one from central west (Cuiabá) and two from northeastern (Campina Grande and Teresina) regions of Brazil. Genetic distances (pairwise values of FST and Nm) and the analysis of molecular variance (AMOVA) were statistically significant, independent of geographic distances among the sites analyzed, indicating that them are under a complex dynamic process that influence the levels of gene flow within and among regions of the country. Bayesian analysis in STRUCTURE revealed the existence of two major genetic clusters, as well as there was genetic substructure within them; these results were confirmed by AMOVA, BAPS and DAPC analyses. This differentiation is the cumulative result of several factors combined as events of multiple introduction, passive dispersal, environmental and climatic conditions, use of insecticides, cycles of extinction and re-colonization followed by microevolutionary processes throughout the country. Isolation by distance also contributed to this differentiation, especially among geographically closer localities. These genetic differences may affect its vector competence to transmit dengue, chikungunya, Zika and the response to vector control programs.

Epidemiological relevance of dogs for the prevention of Toxoplasma gondii, Neospora caninum and Leptospira spp.

This study evaluated the seroprevalence of Toxoplasma gondii, Neospora caninum and Leptospira spp. in dogs from Foz do Iguaçu, Paraná, Brazil. Indirect immunofluorescent antibody test was used to detect antibodies anti-T.gondii and anti-N. caninum. Immunoenzymatic assay and microscopic serum agglutination were used for screening antibodies anti-T.gondii and anti-Leptospira spp., respectively. The results were: 67.02% of the samples reactive for T.gondii and 1.38% for N. caninum, both without statistically significant variables. For Leptospira spp. the results indicated seroprevalence of 23.11%. The analysis of the variables without distinction of serovar showed association for intrinsic characteristics as breed, age, nutritional status and dog category. The extrinsic variables as city region and access to the street presented association (p<0.05). The most prevalent serovars were: Canicola 59.47%; Bratislava 13.07% and Butembo 15.68%. Variables that make up the adjusted multiple analysis model using Leptospira spp. were: age, breed and nutritional status; serovar Canicola, sex, nutritional status and area (p<0.05); serovar Bratislava, lymphadenomegaly and presence of fleas (p<0.05). Given the results obtained, dogs can be used as sentinels for toxoplasmosis and leptospirosis in Foz do Iguaçu and other cities with similar outcomes. In addition, preventive measures should be taken by health authorities because they are zoonoses and humans are also at risk.

Tick-borne pathogens in carthorses from Foz do Iguaçu City, Paraná State, southern Brazil: A tri-border area of Brazil, Paraguay and Argentina.

Tick-borne diseases (TBD) constitute an important group of illness affecting animals and humans worldwide. In Brazil, carthorses are frequently exposed to ticks and tick-borne pathogens, leading to impairment of horse performance and imposing restrictions by the international veterinary authorities for the importation of horses. Accordingly, this study has aimed to i) determine the prevalence of the TBD agents Theileria equi, Babesia caballi, Ehrlichia spp., and hemotropic mycoplasmas in carthorses, ii) identify the tick species parasitizing the animals, and iii) determine factors associated with exposure/infection in Foz do Iguaçu City, Parana state, southern Brazil. A total of 103 carthorses were screened for anti-T. equi and anti-Ehrlichia spp. antibodies by indirect fluorescent antibody assays (IFA). Samples were also tested by PCR assays targeting the 18S rRNA gene of T. equi and B. caballi, and 16S rRNA gene of hemoplasmas. Additionally, PCR assays targeting the 16S rRNA, disulfide bond formation protein (dsb) and tandem repeat proteins 36 (trp36) genes of Ehrlichia spp. were also performed. Antibodies to T. equi and Ehrlichia spp. were detected in 43/103 (41.75%; 95% CI: 32.10-51.88%) and 5/103 (4.85%; 95% CI: 1.59-10.97%) horses by IFA, respectively. DNA of T. equi and B. caballi were found in 25/103 (24.27%; 95% CI: 16.36-33.71%) and 10/103 (9.71%; 95% CI: 4.75-17.13%) carthorses, respectively, and all tested negative for Ehrlichia spp. and hemoplasmas. All sequences showed ≥99% identity with multiple T. equi and B. caballi 18S rRNA gene sequences deposited in GenBank. Overall, 191 Dermacentor nitens ticks were collected from 25/103 (24.27%) animals. Carthorses older than 5 years were more likely to be positive for T. equi (p < 0.05). In conclusion, equine piroplasmosis agents are highly prevalent in carthorses from Foz do Iguaçu City. The low prevalence of Ehrlichia spp. found may be due to the absence of Amblyomma ticks infesting animals, which should be further investigated.

Building Infestation Index for Aedes aegypti and occurrence of dengue fever in the municipality of Foz do Iguaçu, Paraná, Brazil, from 2001 to 2016.

INTRODUCTION: the Building Infestation Index (BII) uses the Rapid Assay of the Larval Index for Aedes aegypti (LIRAa) to express the relationship between positive and surveyed properties. We evaluated LIRAa and the relationship between the BII and climate variables for dengue cases in Foz do Iguaçu municipality, Paraná. METHODS: Spearman's correlations for mean precipitation, mean temperature, BII, and dengue cases (time lag). RESULTS: positive correlations between BII and cases, and mean temperature and cases at two months. Weak correlation between precipitation and cases at three months. CONCLUSIONS: LIRAa and climate variables correlate with dengue cases.

Hidden danger: Unexpected scenario in the vector-parasite dynamics of leishmaniases in the Brazil side of triple border (Argentina, Brazil and Paraguay).

Every year about 3 million tourists from around the world visit Brazil, Argentina and Paraguay´s triple border region where the Iguaçu Falls are located. Unfortunately, in recent years an increasing number of autochthonous canine and human visceral leishmaniasis (VL) cases have been reported. The parasite is Leishmania (Leishmania) infantum and it is transmitted by sand flies (Phlebotominae). To assess the risk factors favorable for the establishment and spread of potential vectors the Centers for Disease Control and Prevention light trap (CDC-light trap) collections were made in the Foz do Iguaçu (FI) and Santa Terezinha de Itaipu (STI) townships and along two transects between them. Our study determined the Phlebotominae fauna, the factors that affect the presence and abundance of Lutzomyia longipalpis and Nyssomyia whitmani, the presence of L. infantum in different sand fly species and which Leishmania species are present in this region. Lutzomyia longipalpis was the prevalent species and its distribution was related to the abundance of dogs. Leishmania infantum was found in Lu. longipalpis, Ny. whitmani, Ny. neivai and a Lutzomyia sp. All the results are discussed within the Stockholm Paradigm and focus on their importance in the elaboration of public health policies in international border areas. This region has all the properties of stable VL endemic foci that can serve as a source of the disease for neighboring municipalities, states and countries. Most of the urban areas of tropical America are propitious for Lu. longipalpis establishment and have large dog populations. Pan American Health Organization´s initiative in supporting the public health policies in the border areas of this study is crucial and laudable. However, if stakeholders do not act quickly in controlling VL in this region, the scenario will inevitable become worse. Moreover, L. (Viannia) braziliensis found in this study supports the need to develop public health policies to avoid the spread of cutaneous leishmaniasis. The consequences of socioeconomic attributes, boundaries and frontiers on the spread of diseases cannot be neglected. For an efficient control, it is essential that urban planning is articulated with the neighboring cities.

[Specificity of the Adultrap for capturing females of Aedes aegypti (Diptera: Culicidae)]. / Especificidade da armadilha Adultrap para capturar fêmeas de Aedes aegypti (Diptera: Culicidae).

The Adultrap is a new trap built for capturing females of Aedes aegypti. Tests were carried out to evaluate the specificity of this trap in comparison with the technique of aspiration of specimens in artificial shelters. Adultraps were kept for 24 hours inside and outside 120 randomly selected homes in two districts of the city of Foz do Iguaçú, State of Paraná. The statistical test was Poissons log-linear model. The result was 726 mosquitoes captured, of which 80 were Aedes aegypti. The Adultrap captured only females of this species, while the aspiration method captured both sexes of Aedes aegypti and another five species. The Adultrap captured Aedes aegypti inside and outside the homes, but the analysis indicated that, outside the homes, this trap captured significantly more females than aspiration did. The sensitivity of the Adultrap for detecting females of Aedes aegypti in low-frequency situations was also demonstrated.