Morphological Description of the Female, Pupa, and Fourth-Instar Larva and Redescription of the Male of Culex (Melanoconion) aliciae (Diptera: Culicidae).

The female, pupa, and fourth-instar larva of Culex (Melanoconion) aliciae are described for the first time, and the male is redescribed. The cibarium, female and male genitalia, pupa, and fourth-instar larva are illustrated. Information on distribution, bionomics, and taxonomy is also included. Males of Cx. aliciae Duret and Cx. martinezi Casal & Garcia are indistinguishable, except for the appearance of the ninth tergal lobes. The female of Culex martinezi has not been described before. The females of Culex aliciae can be recognized by characteristics of the occipital scales, cibarium, and genitalia. The larva of Cx. aliciae and Cx. martinezi can be distinguished from characters of the thorax and abdomen.

Ecological Characterization of Mosquitoes (Diptera: Culicidae) at the Southern Coast of Mar Chiquita Lake, Argentina.

In the southern coast of Mar Chiquita Lake, central Argentina, mosquitoes affect public health and community livelihood, since they transmit pathogens to human beings causing diseases such as malaria, filariasis, encephalitis, yellow fever, and dengue, among others, and have a negative effect on cattle farming as well. To characterize the structure of the mosquito assemblage of the region, we determined the species composition and diversity, the temporal distribution of different species, and the patterns of species richness, abundance, and diversity across seasons. We collected adult mosquitoes over a two-year period (October 2004-September 2006) by means of CDC light traps baited with CO2 from 18:00 to 08:00 h during the warm season (October-April) and from 12:00 h to 18:00 h in the cold season (May-September). A total of 71,501 individuals from 30 species were collected, with Culex Linnaeus and Aedes Meigen genera representing more than 98% of collected specimens (61.5% and 37.3%, respectively). The higher values of richness and abundance of Culicidae were registered in warm seasons compared to cold seasons. Chao1 estimates suggested that more than 90% of the species were detected in all seasons. Mosquito abundance distribution fit the logarithmic series and log-normal models. Aedes albifasciatus (Macquart), Ae. scapularis (Rondani), Culex interfor Dyar, Cx. saltanensis Dyar, and Cx. dolosus (Lynch Arribálzaga), vectors incriminated in arbovirus transmission, were abundant year-round, with Cx. saltanensis and Cx. dolosus most prevalent in cold seasons. Further studies are needed to assess the role of these species in arbovirus transmission in this region of central Argentina.

Understanding the role of temporal variation of environmental variables in predicting Aedes aegypti oviposition activity in a temperate region of Argentina.

Environmental variables related to vegetation and weather are some of the most influential factors that impacting Aedes (Stegomya) aegypti, a mosquito vector of dengue, chikungunya and Zika viruses. In this paper, we aim to develop temporal predictive models for Ae. aegypti oviposition activity utilizing vegetation and meteorological variables as predictors in Córdoba city (Argentina). Eggs were collected using ovitraps placed throughout the city from 2009 to 2012 that were replaced weekly. Temporal generalized linear mixed models were developed with negative binomial distributions of errors that model average number of eggs collected weekly as a function of vegetation and meteorological variables with time lags. The best model included a vegetation index, vapor pressure of water, precipitation and photoperiod. With each unit of increment in vegetation index per week the average number of eggs increased by 1.71 in the third week. Furthermore, each millimeter increase of accumulated rain during 4 weeks was associated with a decrease of 0.668 in the average number of eggs found in the following week. This negative effect of precipitation could occur during abundant rainfalls that fill containers completely, thereby depriving females of oviposition sites and leading them to search for other suitable breeding sites. Furthermore, the average number of eggs increased with the photoperiod at low values of mean vapor pressure; however the average number of eggs decreased at high values of mean vapor pressure, and the positive relationship between the response variable and mean vapor pressure was stronger at low values of photoperiod. Additionally, minimum temperature was associated positively with oviposition activity and that low minimum temperatures could be a limiting factor in Ae. aegypti oviposition activity. Our results emphasize the important role that climatic variables such as temperature, precipitation, and vapor pressure play in Ae. aegypti oviposition activity and how these variables along with vegetation indices can be used to inform predictive temporal models of Ae. aegypti population dynamics that can be used for informing mosquito population control and arbovirus mitigation strategies.

Autochthonous circulation of Saint Louis encephalitis and West Nile viruses in the Province of La Rioja, Argentina.

St. Louis encephalitis (SLEV) and West Nile (WNV) arboviruses, which circulate in Argentina, are maintained in enzootic transmission cycles involving Culex mosquitoes (vectors) and birds belonging to orders Passeriformes and Columbiformes (amplifier hosts). The objective of this work was to determine the circulation of both viruses among wild birds in a semiarid ecosystem in the Province of La Rioja through a serologic survey. During spring 2013 and fall 2014, a total of 326 wild birds belonging to 41 species were captured in areas close to the cities of La Rioja and Chilecito, in the Province of La Rioja. While exposure to SLEV and WNV was analyzed in birds' serum through neutralizing antibody detection, viral circulation was estimated through apparent seroprevalence of neutralizing antibodies. The exposure of the avian community to viruses was 3.02% for SLEV and 1.89% for WNV, while 1.19% corresponded to coinfections. Our study confirms for the first time the circulation of SLEV and WNV in wild birds in the Province of La Rioja. Moreover, it is the first study to register neutralizing antibodies for flavivirus in the species Leptotila verreauxi (White-tipped Dove) (WNV) and Melanerpes cactorum (White-fronted Woodpecker) (SLEV). These results suggest that in semiarid ecosystems from northwestern Argentina the requirements and conditions for amplification and enzootic maintenance of SLEV and WNV would be present.

A decade of arbovirus emergence in the temperate southern cone of South America: dengue, Aedes aegypti and climate dynamics in Córdoba, Argentina.

BACKGROUND: Argentina is located at the southern temperate range of arboviral transmission by the mosquito Aedes aegypti and has experienced a rapid increase in disease transmission in recent years. Here we present findings from an entomological surveillance study that began in Córdoba, Argentina, following the emergence of dengue in 2009. METHODS: From 2009 to 2017, larval surveys were conducted monthly, from November to May, in 600 randomly selected households distributed across the city. From 2009 to 2013, ovitraps (n = 177) were sampled weekly to monitor the oviposition activity of Ae. aegypti. We explored seasonal and interannual dynamics of entomological variables and dengue transmission. Cross correlation analysis was used to identify significant lag periods. RESULTS: Aedes aegypti were detected over the entire study period, and abundance peaked during the summer months (January to March). We identified a considerable increase in the proportion of homes with juvenile Ae. aegypti over the study period (from 5.7% of homes in 2009-10 to 15.4% of homes in 2016-17). Aedes aegypti eggs per ovitrap and larval abundance were positively associated with temperature in the same month. Autochthonous dengue transmission peaked in April, following a peak in imported dengue cases in March; autochthonous dengue was not positively associated with vector or climate variables. CONCLUSIONS: This longitudinal study provides insights into the complex dynamics of arbovirus transmission and vector populations in a temperate region of arbovirus emergence. Our findings suggest that Córdoba is well suited for arbovirus disease transmission, given the stable and abundant vector populations. Further studies are needed to better understand the role of regional human movement.

Meteorological variables associated with the temporal oviposition rate of Aedes aegypti (Diptera: Culicidae) in Resistencia city, Chaco province, Northeastern Argentina.

Aedes aegypti is the main vector of dengue virus in South America. In the last fifteen years, Argentina has suffered three large dengue outbreaks: one in 2009, another one in 2016 and the current 2020 outbreak, with 26,000, 66,000 and more than 43,000 confirmed cases, respectively. These outbreaks are associated with the circulation of the virus in neighboring countries due to the constant movement of people across the frontier. In 2009, the main province affected was Chaco province, with 11,037 confirmed cases (50% of total cases). Thus, the aim of this study was to characterize the temporal oviposition rate of Aedes aegypti in Resistencia, the capital city of this province, in relation to meteorological variables. Mosquitoes were sampled weekly, from April 2015 to March 2017, using ovitraps located in houses randomly selected within the urban area. Oviposition rate was evaluated by means of the total number of eggs collected per week and the proportion of positive ovitraps per week. The coefficient of variation for these two indicators was estimated for the whole study period and then compared using the Wilcoxon Pair test. A generalized linear model (GLM) was performed to infer association between meteorological variables and oviposition rate without time lag and with a time lag of 1 to 4 weeks, considering the biology of this mosquito species. Comparisons between the coefficient of variation of egg number versus the proportion of positive ovitraps showed significant differences, being the former more variable than the latter. The most significant time lag was 2 weeks for the minimum temperature. The number of Ae. aegypti eggs increased 4.05 times when the minimum temperature increased 1°C two weeks before, and decreased 0.69 times with an increase of 1 unit in relative humidity. No eggs were collected when the temperature was below 8°C. The oviposition rate was associated with the increase in the critical minimum temperature (over 8°C) and the weekly frequency of precipitation and dissociated with relative humidity. The maximum oviposition rate was recorded between November and January of both years. The GLM could not explain the relation between meteorological variables and the proportion of positive ovitraps. The peaks of Ae. aegypti egg abundance occurred simultaneously with dengue autochthonous cases recorded in Chaco province, resulting in a good indicative of the period of greatest epidemiological risk.

Culex interfor and Culex saltanensis (Diptera: Culicidae) are susceptible and competent to transmit St. Louis encephalitis virus (Flavivirus: Flaviviridae) in central Argentina.

BACKGROUND: St. Louis encephalitis virus (SLEV) is endemic and autochthonous on the American continent. Culex pipiens quinquefasciatus is a vector of SLEV; however, Culex interfor and Culex saltanensis have also been found to be naturally infected with SLEV. The aim of this study was to determine the vector competence of C. interfor and C. saltanensis for SLEV from Argentina compared with C. p. quinquefasciatus. METHODS: Female of the Culex species were orally infected by feeding on viraemic chicks that had been inoculated with SLEV. Abdomens, legs and saliva blood-fed mosquitoes were analysed by viral plaque assay. RESULTS: Mosquitoes were susceptible to orally acquired infection, dissemination and transmission of SLEV in the saliva. CONCLUSIONS: Our results demonstrate that C. saltanensis and C. interfor are susceptible to SLEV and competent for its transmission.

Increasing the digital repository of DNA barcoding sequences of sand flies (Psychodidae: Phlebotominae).

Sand fly identification is complex because it depends on the expertise of the taxonomist. The females show subtle morphological differences and the occurrence of the species complexes are usual in this taxon. Therefore, a fragment of the cytochrome c oxidase subunit I (COI) gene is used for taxon barcoding to resolve this kind of problem. This study incorporates barcode sequences, for the first time, for Evandromyia cortelezzii and Migonemyia migonei from Argentina. The nucleotide sequence divergences were estimated to generate a neighbour-joining (NJ) tree. The automatic barcode gap discovery (ABGD) approach was employed to find the barcode gaps and the operational taxonomic unit (OTU) delimitation. Other species of the subtribe were included. The frequency histogram of divergences showed a barcoding gap. The ABGD analysis identified 14 operational taxonomic units (OTUs) from 13 morphological species. Sequences of Ev. cortelezzii and Mg. migonei formed well supported clusters and were diagnosed as primary species. These sequences are useful tools for molecular identification of the sand flies of the New World.

Mosquito (Diptera: Culicidae) larval ecology in natural habitats in the cold temperate Patagonia region of Argentina.

BACKGROUND: Knowledge of immature habitats is an important focus for investigations of mosquito community ecology, and may improve our understanding of how environmental variables increase risk of mosquito-borne diseases by influencing the distributions and abundances of species. In Patagonia region, where climatic and ecological factors could be only borderline suitable for mosquito development, relatively little is known about larval ecology. The present study focuses on associations of environmental conditions in natural aquatic habitats with abundances of mosquito species that have colonized such habitats in Patagonia. METHODS: We described the mosquito community composition within 26 natural temporary pools, and assessed the general relationships between environmental variables (pH, water temperature, conductivity, salinity, dissolved oxygen, aquatic plant cover and main nutrients) and larval abundances using redundancy analysis (RDA). Additionally, we compiled monthly climate data and vegetation indices for each larval habitat, and estimated the probability of presence for two of the most abundant species, describing through generalized linear models (GLM) the environmental, climatic and landscape variables-probability of occurrence relationships. RESULTS: Seven species belonging to the genera Culex and Aedes were identified, with Culex apicinus, Cx. acharistus and Aedes albifasciatus being the most abundant. Mean larval densities were low (6.8 ± 2.8 larvae/dip), and the highest species richness and larval densities were recorded in northern and central areas. Aedes albifasciatus, a species of sanitary importance, was widely distributed, being the only one collected south of the 45th parallel of S latitude. RDA indicated that aquatic conductivity, pH, water depth, dissolved oxygen, ammonia and soluble reactive phosphorous accounted for the main part of the variation in the species composition. According to GLMs, wind speed was the variable that best described the presence of Ae. albifasciatus, and the probability of finding this species was positively associated with high wind speed values. On the other hand, the EVI vegetation index was the only variable included in the Cx. apicinus model, whereby there was a great probability of presence in arid areas with lower EVI values. CONCLUSIONS: Our results enhance our knowledge of larval habitat ecology under the extreme environmental conditions of Patagonia and will guide future efforts to understand how multiple effects can affect mosquito ecology and public health at higher latitudes.

Seasonal distribution of Phlebotomine sandfly in a vulnerable area for tegumentary leishmaniasis transmission in Córdoba, Argentina.

Thirty-seven sandfly species are listed for Argentina distributed in 14 provinces and Leishmaniasis cases extend from the north of the country to Unquillo City (Córdoba Province), but potential vectors are found further to the south. This is the first study on diversity, spatial and temporal distribution of sandflies on the outskirts of the temperate Córdoba City, and the factors that influence their presence. Migonemyia migonei, record here for Córdoba City for the first time, and the Evandromyia cortelezzii-sallesi Complex was found, also Ev. cortelezzii males were captured for the first time, these sandflies being more abundant during the warm months due to meteorological factors and the presence of blood meal sources. At least the eastern outskirts of Córdoba City, the second most important city of the country, are at risk of Leishmaniasis transmission if Leishmania spp. enters into the area due to the presence of competent vectors and adequate vertebrate hosts, in a favorable socio-economic context.

Molecular identification of two Culex (Culex) species of the neotropical region (Diptera: Culicidae).

Culex bidens and C. interfor, implicated in arbovirus transmission in Argentina, are sister species, only distinguishable by feature of the male genitalia; however, intermediate specimens of the species in sympatry have been found. Fourth-instar larvae and females of both species share apomorphic features, and this lack of clear distinction creates problems for specific identification. Geometric morphometric traits of these life stages also do not distinguish the species. The aim of the present study was to assess the taxonomic status of C. bidens and C. interfor using two mitochondrial genes and to determine the degree of their reproductive isolation using microsatellite loci. Sequences of the ND4 and COI genes were concatenated in a matrix of 993 nucleotides and used for phylogenetic and distance analyses. Bayesian and maximum parsimony inferences showed a well resolved and supported topology, enclosing sequences of individuals of C. bidens (0.83 BPP, 73 BSV) and C. interfor (0.98 BPP, 97 BSV) in a strong sister relationship. The mean K2P distance within C. bidens and C. interfor was 0.3% and 0.2%, respectively, and the interspecific variation was 2.3%. Bayesian clustering also showed two distinct mitochondrial lineages. All sequenced mosquitoes were successfully identified in accordance with the best close match algorithm. The low genetic distance values obtained indicate that the species diverged quite recently. Most morphologically intermediate specimens of C. bidens from Córdoba were heterozygous for the microsatellite locus GT51; the significant heterozygote excess observed suggests incomplete reproductive isolation. However, C. bidens and C. interfor should be considered good species: the ventral arm of the phallosome of the male genitalia and the ND4 and COI sequences are diagnostic characters.

Weather Variability Associated with Aedes (Stegomyia) aegypti (Dengue Vector) Oviposition Dynamics in Northwestern Argentina.

This study aims to develop a forecasting model by assessing the weather variability associated with seasonal fluctuation of Aedes aegypti oviposition dynamic at a city level in Orán, in northwestern Argentina. Oviposition dynamics were assessed by weekly monitoring of 90 ovitraps in the urban area during 2005-2007. Correlations were performed between the number of eggs collected weekly and weather variables (rainfall, photoperiod, vapor pressure of water, temperature, and relative humidity) with and without time lags (1 to 6 weeks). A stepwise multiple linear regression analysis was performed with the set of meteorological variables from the first year of study with the variables in the time lags that best correlated with the oviposition. Model validation was conducted using the data from the second year of study (October 2006- 2007). Minimum temperature and rainfall were the most important variables. No eggs were found at temperatures below 10 °C. The most significant time lags were 3 weeks for minimum temperature and rains, 3 weeks for water vapor pressure, and 6 weeks for maximum temperature. Aedes aegypti could be expected in Orán three weeks after rains with adequate min temperatures. The best-fit forecasting model for the combined meteorological variables explained 70 % of the variance (adj. R(2)). The correlation between Ae. aegypti oviposition observed and estimated by the forecasting model resulted in rs = 0.80 (P < 0.05). The forecasting model developed would allow prediction of increases and decreases in the Ae. aegypti oviposition activity based on meteorological data for Orán city and, according to the meteorological variables, vector activity can be predicted three or four weeks in advance.

St. Louis Encephalitis virus mosquito vectors dynamics in three different environments in relation to remotely sensed environmental conditions.

In Argentina the St. Louis Encephalitis virus (SLEV) is an endemic and widely distributed pathogen transmitted by the cosmopolitan mosquito Culex quinquefasciatus. During two outbreaks in Córdoba city, in 2005 and 2010, Culex interfor was also found infected, but its role as vector of SLEV is poorly known. This mosquito species is distributed from central Argentina to southern Brazil. The primary aim of this study was to analyze the population dynamic of Cx. interfor and Cx. quinquefasciatus in three different environments (urban, suburban and non-urban) in relation to remotely sensed environmental data for vegetation (NDVI and NDWI) and temperature (brightness temperature). Cx. quinquefasciatus and Cx. interfor were found at the three sampled sites, being both the most abundant Culex species, with peaks in early and midsummer. Temporal distribution patterns of both mosquito species were highly correlated in a non-urban area of high SLEV risk transmission. Cx. quinquefasciatus and Cx. interfor were associated with the most urbanized site and the non-urban environment, respectively; high significant correlations were detected between vegetation indices and abundance of both mosquito species confirming these associations. These data provide a foundation for building density maps of these two SLEV mosquito vectors using remotely sensed data to help inform vector control programs.

Effect of temperature on life history traits during immature development of Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae) from Córdoba city, Argentina.

We investigated how ambient temperature under fluctuating conditions affects the larval-pupal immature traits of Aedes aegypti and Culex quinquefasciatus mosquitoes from Córdoba city, Argentina, and established each species development threshold and physiological time. Based on life tables, three cohorts of each mosquito species were reared in the laboratory under small fluctuating temperatures conditions of 15.2±1.7°C, 17.9±1.6°C, 21.6±0.7°C and 25.3±0.4°C for Ae. aegypti, and 16.6±1.7°C, 18.7±1.7°C and 25.2±0.3°C for Cx. quinquefasciatus. Immature development time and survival values, and also thermal development threshold and physiological time were estimated. Development times of all larval and pupal stages of Ae. aegypti and Cx. quinquefasciatus were significantly affected by the rearing temperatures, decreasing when temperature increased. Mean Ae. aegypti total (larva+pupa) development time ranged from 21.9 to 8.6 days, at 15.2 and 25.3°C, whereas, for Cx. quinquefasciatus varied between 23.5 to 9.2 days at 16.6 and 25.2°C, respectively. Larval and pupal survival of both species was affected by different rearing temperatures, increasing in general as temperature increased. For Ae. aegypti the total immature survival ranged from 26% at 15.2°C to 92% at 21.6°C; however, temperature did not have significant effect on this variable. The total immature survival of Cx. quinquefasciatus was significantly and positively affected by temperatures, ranging from 32 to 88%, at 16.6 and 25.2°C. The temperature development threshold and the physiological time estimated for Ae. aegypti and Cx. quinquefasciatus were 11.11°C and 93.74 degree-days, and 10.96°C and 136.87 degree-days, respectively. The results of the present study showed that temperature significantly affects the larval-pupal immature traits of these mosquito species of sanitary importance, from the central region of Argentina. All the parameters recorded are useful for the development of mosquito management models.

Demographic history and population structure of Anopheles pseudopunctipennis in Argentina based on the mitochondrial COI gene.

BACKGROUND: Anopheles pseudopunctipennis is an important malaria vector in the Neotropical region and the only species involved in Plasmodium transmission in the Andean foothills. Its wide geographical distribution in America, high preference for biting humans and capacity to rest inside dwellings after feeding, are attributes contributing to its vector status. Previous reports have tried to elucidate its taxonomic status, distinguishing populations from North, Central and South America. In the present study we used a mitochondrial marker to examine the demographic history of An. pseudopunctipennis in northwestern Argentina. METHODS: Twelve localities were selected across 550 km of the distribution of this species in Argentina, including two near the Bolivian border and several in South Tucumán, for sampling. A fragment of the cytochrome oxidase I (COI) gene was sequenced and haplotype relationships were analyzed by a statistical parsimony network and a Neighbor-Joining (NJ) tree. Genetic differentiation was estimated with FST. Historical demographic processes were evaluated using diversity measures, neutrality tests and mismatch distribution. RESULTS: Forty-one haplotypes were identified, of which haplotype A was the most common and widely distributed. Neither the network nor the NJ tree showed any geographic differentiation between northern and southern populations. Haplotype diversities, Tajima's DT and Fu & Li's F and D neutrality tests and mismatch distribution supported a scenario of Holocene demographic expansion. CONCLUSION: The demographic pattern suggests that An. pseudopunctipennis has undergone a single colonization process, and the ancestral haplotype is shared by specimens from all localities, indicating mitochondrial gene flow. Genetic differentiation was minimal, observed only between one northern and one southern locality. The estimated time of the population expansion of this species was during the Holocene. These data suggest that regional vector control measures would be equally effective in both northern and southern localities sampled, but also that insecticide resistant genes may spread rapidly within this region.

Phylogenetic relationships of Culex (Culex) species (Diptera, Culicidae) from Argentina based on morphological characters.

The purpose of this study was for the first time to establish phylogenetic hypotheses among Culex (Culex) species from Argentina, and to evaluate the relationships of this subgenus with subgenera Phenacomyia and Phytotelmatomyia. We employed maximum parsimony to analyse morphological characters derived from the morphology of the adult, pupal and larval stages. More than 30 figures, including photographs, illustrating 87 characters are provided. The cladistic analysis of 61 terminal taxa and 95 characters was undertaken, under implied weighting method, with a range of K values from 6 to 9. Three unstable taxa were detected therefore a reduced strict consensus tree was created. The subgenus Culex appears polyphyletic relative to subgenus Oculeomyia. The internal classification of subgenus Culex is shown to be chaotic; the Pipiens Group is polyphyletic relative to the Sitiens and Coronator Groups, the latter being polyphyletic with respect to the Pipiens Group. Phylogenetic analysis combining characters provided by morphology and molecular sources are needed to interpret more fully the relationships in the group.

New records of mosquito species in the provinces of Chaco and Formosa, Argentina.

Seven mosquito species are recorded for the 1st time for northeastern Argentina: Culex (Melanoconion) albinensis, Cx. (Mel.) elevator, Cx. (Mel.) intrincatus, and Cx. (Mel.) serratimarge for Formosa Province, and Sabethes (Peytonulus) undosus, Sa. (Sabethinus) melanoninphe, and Toxorhynchites (Lynchiella) theobaldi for Chaco Province. The geographical distribution of these species is extended to northeastern Argentina, and the number of species increases to 97 and 75 for the provinces of Chaco and Formosa, respectively.

Mapping environmental susceptibility to Saint Louis encephalitis virus, based on a decision tree model of remotely-sensed data.

In response to the first human outbreak (January May 2005) of Saint Louis encephalitis (SLE) virus in Córdoba province, Argentina, we developed an environmental SLE virus risk map for the capital, i.e. Córdoba city. The aim was to provide a map capable of detecting macro-environmental factors associated with the spatial distribution of SLE cases, based on remotely sensed data and a geographical information system. Vegetation, soil brightness, humidity status, distances to water-bodies and areas covered by vegetation were assessed based on pre-outbreak images provided by the Landsat 5TM satellite. A strong inverse relationship between the number of humans infected by SLEV and distance to high-vigor vegetation was noted. A statistical non-hierarchic decision tree model was constructed, based on environmental variables representing the areas surrounding patient residences. From this point of view, 18% of the city could be classified as being at high risk for SLEV infection, while 34% carried a low risk, or none at all. Taking the whole 2005 epidemic into account, 80% of the cases came from areas classified by the model as medium-high or high risk. Almost 46% of the cases were registered in high-risk areas, while there were no cases (0%) in areas affirmed as risk free.