Figure 3 illustrates households communities visits, applying a survey to the house-hold leader (Fig. 3a), as well as cataloging breeding sites around the property (Figs. 3b to 3f), which are favorable to Culicidae proliferation.
The breeding sites found in the peridomestic environments (Fig. 3) were similar to those reported in the literature when associated with basic sanitation (Table 1). Tire presence represented 26% of the total frequency, as the most frequent container, followed by plastic drums (22%), and water tank and animal water troughs, both with 14%. These results corroborate Ngugi et al (2017) found, who reported that the most frequent types of habitat were buckets, drums, tires and pots, representing more than 75% of all pupae, concluding that efforts to control Culicidae population could be directed towards eliminating these breeding sites, being a way cost-effective way to reduce arboviral transmission.
Table 1
Breeding sites with suspicious forms of Aedes aegypti occurrence on rural communities in Goiás, Brazil, compared with data from literature
Observed in this research (in situ) | Literature data |
Local | Occurrence (%) | Local¹ | Reference |
Plastic drums | 22.0 | Large reservoirs that are poorly or uncovered: water tanks, gallons, casks. | (Guagliardo et al. 2014; Ngugi et al. 2017; IOC/Fiocruz 2017; Rahman et al. 2021) |
Water tank (Fig. 3b) | 14.0 |
Bucket | 12.0 |
Can | 2.0 |
Animal water troughs (Fig. 3d) | 14.0 | Containers for watering animals, among others | (Brazil 2013; Dalpadado, Amarasinghe and Gunathilaka 2022) |
Tire (Fig. 3e) | 26.0 | Accumulated and/or scattered solid waste; organic waste outdoors. | (Lopes et al. 2004; Troyes, Vellegas and Troyes 2006; Manrique-Saide et al. 2008; Ngugi et al. 2017; IOC/Fiocruz 2017; Dieng et al. 2018; Silva et al. 2020; Rahman et al. 2021) |
Pan with rain water | 4.0 |
Bottle and scattered waste (Fig. 3c) | 2.0 |
Rudimentary well (Fig. 3f) | 4.0 | Rudimentary wells, poorly or uncovered. | (Barrera et al. 2008; Mackay et al. 2009; Somers et al. 2011) |
Total | 100.0 | | |
Note: 1 = Breeding sites identified in rural and/or urban areas via inspection of suspicious immature (egg, larva, pupa) and adult forms of Aedes aegypti, considering potential breeding sites related with environmental variables in terms of basic sanitation. |
Complementing Fig. 3 and Table 1, Fig. 4 illustrates the breeding sites with their percentages found in the present research. Among breeding sites, tire is an important container for Culicidae oviposition. This was evidenced in the present research, where immature individuals of suspected Aedes were observed, mainly in discarded tires (26.0%), corroborating the literature.
In this context, Lopes et al (2004) aiming to analyze Aedes aegypti and Aedes albopictus spatial dispersion between rural and urban areas in Paraná (Brazil), verified that tires contain the largest amounts of Culicidae larvae, and both were dispersed in rural area, and recommend resizing control areas, as rural communities can be refuge-like for these vectors. Additionally, other variety of water containers (Bucket – 12.0%; Can – 2.0%; Pan with rain water – 4.0%; Bottle and scattered waste – 20% and Rudimentary well – 4.0%) appear as potential Aedes breeding sites, contributing to its greater or lesser spread in rural and urban areas. Besides that, environment influences Aedes aegypti and Aedes albopictus abundance, with Aedes albopictus being predominant in rural areas in relation to Aedes aegypti (Braks et al. 2003). However, contrary results can be found in literature (Djiappi-Tchamen et al. 2021). Ricas-Rezende (2020) observation is considered the first report of Aedes albopictus infected by dengue and Zika viruses in Brazilian rural areas, specifically in Espírito Santo state. The population of this vector is growing in the Americas (Garcia-Rejon et al. 2021) and is present, already, in all Brazilian states, with emphasis on the Southeast and Midwest regions (Variza et al. 2022).
In Brazilian Midwest, especially in the rural communities studied in the present work, the containers are more susceptible to prolonged water storage due to rain frequency, combined with the lack of proper handling of disposable debris by residents. Dalpadado, Amarasinghe and Gunathilaka (2022) reported that artificial breeding sites represent 80% of the places found with Aedes larvae. Kroth et al (2019) evidenced Culicidae strong preference for oviposition in outdoor containers, with preference of those with higher organic matter concentrations and, according to the authors, it helps to a better understanding of Aedes aegypti for in-habiting anthropogenic environments, being possible to verify by nutrients availability analysis and breeding sites.
The contact of Aedes aegypti eggs with water is the fundamental stimulus for its development cycle (Silva et al. 1993). Basic sanitation absence or precariousness allows all these arboviruses vectors proliferation (Almeida, Cota and Rodrigues 2020), which was observed in the analyzed communities here, in greater or lesser quantities. According to residents, the main motivation for storing water, for different reasons and long-term use, is due to difficulties of access to it. Other Culicidae breeding sites are reported in the literature, such as rain drains and manholes (Paploski et al. 2016); however, they were not identified in the present study, due to the lack of basic sanitation facilities.
Table 2 presents a summary of peridomiciliary aspects, obtained from supplementary material, and seroprevalence data for SDENV-1-4, ZIKV and CHIKV, indicating positivity for immunoglobulin G tests (IgG class) antibodies presence, against these agents. Among the 137 households visited, the present research found the average of 3.0 inhabitant/household. In in 46.7% (64/137) of them (Table 2), at least one person had already been infected at some point in their life with one of the serotypes of SDENV-1-4, ZIKV and/or CHIKV, corresponding to 21.7% (89/411) of the residents, where total seroprevalences were 29.0% for SDENV-1-4, 3.9% for ZIKV and 12.9% for CHIKV, in all 13 communities visited. These results showed similar behavior from lower to higher incidence found by Li et al (2021), where they observed prevalence of 38.0% for dengue, 18.0% for Zika and 25.0% for chikungunya.
However, with lower incidence of arboviruses, mainly Zika; Li et al (2021) in their meta-analysis, analyzed 133 papers with 176,001 participants, reporting rural and urban populations seroprevalences, from developed and developing countries, with similar values for dengue and chikungunya obtained in the present research. This support, also, the hypothesis that arbovirus transmission can occur in rural locations as well as in urban areas, especially when there are favorable environmental conditions for vector proliferation.
Table 2
Breeding sites quantity with and without suspicious presence of Aedes aegypti forms and positivity for dengue (SDENV-1-4), Zika (ZIKV) and chikungunya (CHIKV), with respective seroprevalence, in rural communities. Goiás, Brazil.
Community | Visited households | Breeding sites with suspicious Aedes aegypti forms | Favorable places for Aedes reproduction | Positive for DENV, ZIKV and/or CHIKV | Seroprevalence(d) for SDENV-1-4, ZIKV e CHIKV at community level |
n° | n° | % | n° | n° | % |
ID | Res | Y | Q(a) | ID(b) | Y | Q(c) | ID | Res | SDENV-1-4 | UL | IL | ZIKV | UL | IL | CHIKV | UL | IL |
1 | 8 | 24 | 1 | 3 | 12.5 | 8 | 25 | 5 | 7 | 49.0 | 26.0 | 72.4 | 0.0 | 0.0 | 0.0 | 17.9 | 6.9 | 39.1 |
2 | 10 | 30 | 6 | 8 | 60.0 | 8 | 32 | 9 | 17 | 82.4 | 59.0 | 93.8 | 5.9 | 0.7 | 35.2 | 4.2 | 0.9 | 18.4 |
3 | 11 | 33 | 2 | 2 | 18.2 | 8 | 20 | 2 | 3 | 11.4 | 2.7 | 37.1 | 0.0 | 0.0 | 0.0 | 11.4 | 2.7 | 37.1 |
4 | 7 | 21 | 1 | 2 | 14.3 | 7 | 16 | 1 | 1 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 20.8 | 3.9 | 62.8 |
5 | 13 | 39 | 3 | 3 | 23.1 | 11 | 44 | 8 | 12 | 64.6 | 33.6 | 86.8 | 24.9 | 4.5 | 70.1 | 14.2 | 2.7 | 49.4 |
6 | 6 | 18 | 0 | 0 | 0.0 | 6 | 23 | 5 | 6 | 63.7 | 29.8 | 87.9 | 23.4 | 3.9 | 69.8 | 12.3 | 2.9 | 40.0 |
7 | 10 | 30 | 6 | 6 | 60.0 | 10 | 38 | 2 | 3 | 6.4 | 1.3 | 26.2 | 0.0 | 0.0 | 0.0 | 2.6 | 0.3 | 19.4 |
8 | 19 | 57 | 6 | 6 | 31.6 | 19 | 83 | 7 | 7 | 24.4 | 7.6 | 55.7 | 0.0 | 0.0 | 0.0 | 21.8 | 6.5 | 52.5 |
9 | 9 | 27 | 2 | 5 | 22.2 | 9 | 49 | 6 | 11 | 45.2 | 23.6 | 68.7 | 4.1 | 0.7 | 21.3 | 14.1 | 5.2 | 37.0 |
10 | 11 | 33 | 4 | 4 | 36.4 | 10 | 25 | 8 | 11 | 49.4 | 27.0 | 72.1 | 24.9 | 8.5 | 54.1 | 26.6 | 11.5 | 50.3 |
11 | 11 | 33 | 1 | 1 | 9.1 | 11 | 27 | 6 | 6 | 15.7 | 5.1 | 39.0 | 0.0 | 0.0 | 0.0 | 2.0 | 0.3 | 12.2 |
12 | 11 | 33 | 5 | 8 | 45.4 | 9 | 20 | 4 | 4 | 12.2 | 3.7 | 33.3 | 9.1 | 2.3 | 30.1 | 6.7 | 1.8 | 21.5 |
13 | 11 | 33 | 2 | 2 | 18.2 | 11 | 36 | 1 | 1 | 31.6 | 13.8 | 57.1 | 1.7 | 0.2 | 13.5 | 19.7 | 6.0 | 48.5 |
T | 137 | 411 | 39 | 50 | 28.5 | 127 | 438 | 64 | 89 | 29.0 | 3.9 | 12.9 |
Note: Communities: Canabrava = 1; Extrema = 2; Itajá II = 3; João de Deus = 4; Lageado = 5; Landi = 6; Pelotas = 7; Rochedo = 8; Fazenda Santo Antônio da Laguna = 9; São Lourenço = 10; São Sebastião da Garganta = 11; Sumidouro = 12; Taquarussu = 13; Household identification = ID; Upper limit = UP; Inferior limit = IL; residents quantity = Res; breeding sites quantity (Q) independent of the breeding site type = (a); quantity households with suspicious breeding sites (%) = (b); Breeding site favorable quantity by community = (c); Yes quantity = Y; dengue = DENV; Zika = ZIKV; chikungunya = CHIKV; seroprevalence - secondary data extracted from Pagotto et al (2022) = (d). |
Regarding the possibility of finding breeding sites, their presence was observed at 92.7% (127/137) of visited households. However, 39 of them (28.5%), belong to 92.3% (12/13) of the studied communities, where 50 containers were found containing Culicidae living forms, characterized as suspicious Aedes aegypti in different stages (larvae, pupae and adult form). It is important to note that this situation was not found only at Landi community (6). Seroprevalence of 28.5% is similar to 29.4% found by Ruiz-Díaz et al (2017) in rural communities of the Caribbean coast.
Figure 5 shows breeding sites quantity with suspicious Aedes aegypti live forms (larvae, pupae and/or adult form) of households peridomicile of each community, comparing it to the number of potential breeding sites for culicid vectors reproduction, under the same environment. In 46.1% of the studied communities (6/13), BSSAaegypti was higher in relation to the PPBS, particularly two communities: Extrema (2) and Sumidouro (12) (Fig. 5).
Table 3 presents Pearson's correlation coefficients with statistical significance. Pearson coefficients showed positive relationship (p < 0,05) between BSSAaegypti and PPBS, at household (PPBSH) and community (PPBSC) levels. In an analysis of Table 3 in parallel with Table 2, Extrema (2) community was the one with the highest number of individuals affected by dengue (82.4%), with 85.7% (6/7) of the related variables significantly (0.61 ≤ "r" _"Pearson" ≤ 1.0), highlighting the relationships between SD_Z_C and SD, and with PPBSH, "r" _"Pearson" = 1.0 and "r" _"Pearson" = 0.67, respectively.
Among the 46.1% (6/13) PPBSC (Supplementary material and Fig. 5), 66.7% (4/6) of them showed relationship (p < 0.05) between PPCD, PPCC and/or BSSAaegypti) and SD_Z_C. On Table 3 can be highlighted that PPBSC x SD_Z_C [p < 0,05; 0.81] in Sumidouro; PPBSC x SD_Z_C [p < 0,05; 0.72] in Pelotas and BSSAaegypti x SD_Z_C [p < 0,05; -0.76] in Fazenda Santo Antônio da Laguna.
Table 3
Relationship between seroprevalence for dengue and/or Zika and/or chikungunya, and the number of breeding sites for culicid vectors, presented by Pearson's correlation coefficient.
RC | SD_Z_C | PPBSH | PPBSC | BSSAaegypti |
SD | SC | SD_Z_C | SD | SD_Z_C | SD | SC | PPBSH | SD_Z_C | SD | SC | PPBSH | PPBSC |
\({\text{r}}_{\text{Pearson}}\) | \({\text{r}}_{\text{Pearson}}\) | \({\text{r}}_{\text{Pearson}}\) | \({\text{r}}_{\text{Pearson}}\) |
1 | - | 0.77** | - | - | - | - | - | - | - | - | - | - | - |
2 | 1.00*** | - | 0.67** | 0.67** | - | - | - | 0.67** | - | - | - | 0.61* | 0.66** |
3 | 0.67** | 1.00*** | - | - | - | - | - | 0.62** | - | - | - | - | 0.57* |
4 | - | 1.00*** | - | - | - | - | - | - | - | - | - | - | - |
5 | 0.85*** | - | - | - | -0.55** | -0.66** | - | - | - | - | - | - | - |
6 | 1.00*** | - | - | - | - | - | - | - | - | - | - | - | - |
7 | 1.00*** | 0.67** | - | - | 0.72** | 0.72** | 0.69** | - | - | - | - | - | - |
8 | 0.57** | 0.68*** | - | - | - | 0.44* | - | - | 0.65*** | - | 0.48** | - | - |
9 | 1.00*** | - | - | - | - | - | - | - | -0.76** | -0.76** | - | - | - |
10 | 0.56* | - | - | - | - | - | - | 0.53* | - | - | - | - | - |
11 | 0.83*** | - | - | - | - | 0.52* | - | - | - | - | - | - | - |
12 | - | 0.62** | - | - | 0.81*** | - | 0.59* | 0.64** | - | - | - | - | 0.53* |
13 | 1.00*** | - | - | - | - | - | - | - | - | - | - | - | - |
Note: rural community = RC, names are shown in Table 2; breeding sites with suspicious forms (larvae, pupae and/or adult) of Aedes aegypti = BSSAaegypti; presence of possible breeding sites per household = PPBSH; presence of possible breeding sites per community = PPBSC; seroprevalence for dengue = SD; Zika seroprevalence = SZ; seroprevalence for chikungunya = SC; seroprevalence for dengue and/or Zika and/or chikungunya = SD_Z_C; non-significant correlation = (-); significance level: (***) = p-value ≤ 0.01, (**) = p-value ≤ 0.05, (*) = p-value ≤ 0.1. |
In the same context, Table 4 presents the distances (Km) from each community to the nearest urban center, with an average distance of 31.5 km, ranging from 6.7 to 82.8 km (Variation Coefficient = 66%; Standard deviation = 20.9 km). Comparing with the capital of Goiás state, Goiânia (where the research base was located), the average distance was 339.8 km, ranging from 94.6 to 626.2 km (Coefficient of variation (CV) = 51%; Standard deviation = 174.1 km). Household density per community is also shown, with averages, SD and CV of the distances between households, as well as the average and proportions of Culicidae breeding sites and seroprevalence, data used for Cluster analysis.
Besides the fact that those 13 rural communities studied here, demonstrated different typologies and geographical aspects in Goiás, Brazil; they share several common peridomiciles environmental features, capable of maintaining Aedes aegypti life cycle and, consequently, promote arboviral diseases dissemination.
Table 4
Distances between rural communities, the nearest urban center and Goiás state capital (Goiânia), the number of households per km² and of the variables results used in Cluster analyses.
Rural community | Distance (km) | Household/ km² | Distance between households | Variable |
To closest urban center | To state capital (Goiânia) | Average (m) | STD (m) | CV (%) | PPBSC | PPBS | BSSAaegypti | SD | SZ | SC | SD_Z_C |
Average | Breeding sites proportion | Soroprevalence proportion |
1 Canabrava | 56.8 | 429.1 | 2.20 | 278.40 | 392.45 | 141 | 3.13 | 1.00 | 0.13 | 0.25 | 0.00 | 0.50 | 0.63 |
2 Extrema | 6.7 | 515.1 | 43.62 | 43.00 | 40.52 | 94 | 3.20 | 0.80 | 0.60 | 0.90 | 0.10 | 0.10 | 0.90 |
3 Itajá II | 9.5 | 192.4 | 4.03 | 278.20 | 121.57 | 44 | 1.82 | 0.73 | 0.18 | 0.09 | 0.00 | 0.18 | 0.18 |
4 João de Deus | 25.2 | 108.1 | 2.67 | 297.10 | 99.70 | 34 | 2.29 | 1.00 | 0.14 | 0.00 | 0.00 | 0.14 | 0.14 |
5 Lageado | 37.5 | 518.5 | 2.19 | 446.10 | 187.62 | 42 | 3.38 | 0.85 | 0.23 | 0.54 | 0.15 | 0.15 | 0.62 |
6 Landi | 82.8 | 433.5 | 8.68 | 177.40 | 138.12 | 78 | 3.83 | 1.00 | 0.00 | 0.83 | 0.00 | 0.17 | 0.83 |
7 Pelotas | 41.1 | 545.9 | 3.55 | 14.90 | 7.87 | 53 | 3.80 | 1.00 | 0.60 | 0.20 | 0.00 | 0.10 | 0.20 |
8 Rochedo | 16.7 | 94.6 | 2.95 | 354.50 | 70.91 | 20 | 4.37 | 1.00 | 0.32 | 0.16 | 0.00 | 0.21 | 0.37 |
9 Fazenda Santo Antônio da Laguna | 37.8 | 282.9 | 3.17 | 470.80 | 204.68 | 43 | 5.44 | 1.00 | 0.22 | 0.67 | 0.00 | 0.33 | 0.67 |
10 São Lourenço | 42.4 | 279.7 | 3.10 | 346.20 | 134.63 | 39 | 2.27 | 0.91 | 0.36 | 0.45 | 0.27 | 0.27 | 0.73 |
11 São Sebastião da Garganta | 28.8 | 139.8 | 2.10 | 296.60 | 158.59 | 53 | 2.45 | 1.00 | 0.09 | 0.45 | 0.00 | 0.09 | 0.55 |
12 Sumidouro | 14.5 | 251.9 | 1.72 | 206.00 | 202.83 | 98 | 1.82 | 0.82 | 0.45 | 0.09 | 0.09 | 0.18 | 0.36 |
13 Taquarussu | 10.1 | 626.2 | 6.29 | 247.20 | 151.77 | 61 | 3.27 | 1.00 | 0.18 | 0.09 | 0.00 | 0.00 | 0.09 |
Note: sites with suspected Aedes aegypti = BSSAaegypti; proportion of possible breeding sites in the community = PPBS; average of possible breeding sites per household in the community = PPBSC; seroprevalence for dengue = SD; seroprevalence for Zika = SZ; seroprevalence for chikungunya = SC; seroprevalence for dengue, Zika and/or chikungunya = SD_Z_C; standard deviation = STD; coefficient of variation = CV. |
In Fig. 6a, a dendrogram from cluster analysis is presented, where communities are aggregated in four homogeneous clusters, according to the similarity between their characteristics (Fig. 6b). Canabrava (Cluster 1) and Extrema (Cluster 2) communities were isolated according grouping analysis. This might be possible due to their unique characteristics and differences. In Extrema, their population density is a higher (43.62 hh/km²), followed by the Landi (8.68 hh/km²). Extrema, also, showed greater proximity between households, as well as elevated arboviruses dissemination and breeding sites presence, corroborated by its Pearson correlation coefficient. In Canabrava, population density was low (2.2 hh/km²) and it was one of the communities with the lowest number of significant Pearson correlation coefficients.
The other two Clusters (3 and 4) where conformed by 5 and 6 communities, respectively. In relation to Cluster 4, a greater variation prevailed in all parameters, excepting population density (1.72 to 6.29 hh/km², CVg3 = 47.05%; CVg4 = 67, 16%) and the number of BSSAaegypti (0.18 to 0.60, CVg3 = 65.20%; CVg4 = 67.05%). Cluster 4 was confirmed by Rochedo, Fazenda Santo Antônio da Laguna, Lageado, São Lourenço, Landi and São Sebastião da Garganta, communities characterized by the substantial dispersion of households (dmean = 348.60 m, CV = 30.49%) and a notably low breeding sites CV (CV PPBS = 6.91%), including the lowest variation in seroprevalence for the three arboviruses, demonstrating that these parameters are crucial for studies on this topic.
Final considerations
Dengue is the arbovirus that most infected the population at some point in its life, compared to other arboviruses (Zika and chikungunya), and may be associated with the co-circulation of different serotypes of the virus (DENV-1–4) in the Brazilian territory, as well as Aedes aegypti transmissibility efficiency (Brazil 2009; Novaes, Pinto and Marques 2022). One of the ways to mitigate the transmission of these arboviruses is by combating Aedes aegypti, in which different insecticides have been tested against the vector, such as: ursolic acid (Kamatchi et al 2023), emodin (Chinnasamy et al 2023), isolated compounds (Raguvaran et al 2023), among others. However, bioefficacy against the vector can be compromised due to the socio-environmental variables that contribute to mosquito proliferation in different regions. In Brazil favored is vector, either by the adequate environment for its habitat and/or the lack of basic sanitation services universalization (Morgan, Strode and Salcedo-Sora 2021; Novaes, Pinto and Marques 2022), as informed in the literature, mainly in Brazilian rural areas.
The studied data in the present research revealed that in 48.7% of the households (19/39), where infection by dengue, Zika and/or chikungunya was confirmed by seroprevalence, at least one water container with suspected Aedes aegypti was registered. Although one community (Landi) did not contain breeding sites with suspicious Aedes live forms, potential breeding sites were found in 5.3% of the households (23/438), in addition to the expressive seroprevalence rates for the three arboviruses, emphasizing the second highest value for Zika (23.4%) and the third highest value for dengue (63.7%).
In 30.8% of the communities, more than 1/3 of the households showed immatures and adults suspicious Aedes aegypti live forms, including São Lourenço community (10), where the highest seroprevalence for chikungunya was observed (26.6%).
Chikungunya was the only arbovirus that in all 13 communities at least one household had been infected. One of the reasons for this scenario may be due its spread in Brazil, prevailing two species of mosquitoes that transmit the disease: Aedes aegypti and Aedes albopictus (Honório et al. 2015; Souza et al. 2022; Almeida et al. 2022).
It was found in the peridomicile of 73 households, where everyone tested had no previous contact with at least one of the arboviruses mentioned (DENV, ZIKV and CHIKV), 26.0% (19/73), the presence of at least one container with immature and adult suspicious Aedes aegypti live forms. This was observed in 69.2% (9/13) of the communities, which could serve as warning to municipal health authorities. The relationship between the number of breeding sites found (PPBS, PPBSC and/or BSSAaegypti) with at least one of the three arboviruses, was observed in 53.8% (7/13) of the communities (Table 3). Therefore, it can be demonstrated that in a significant portion of the rural communities visited (53.8%), the breeding sites found in peridomicile location were probably Culicidae vector habitat, responsible for infecting part of their population regardless of, whether or not, mosquito identification during this investigation.
Thus, confirming the presence of Aedes aegypti or Aedes albopictus, vectors carrying an arbovirus, after biting an infected person coming mainly from the urban area, at least 25% of the families in at least 69.2% (9/13) of the rural communities studied, can be affected due the possible vector proliferation in rural areas on Brazilian Midwest.
Therefore, better evaluation is recommended, especially regarding entomological focus in order to increase vectors understanding and their dispersion in this region.
This research is limited by the seroprevalence data obtained from late infections, it means that the studied population had previously contact with, at least, one of the arboviruses, at some point in their lives, as a methodology commonly used in other studies (Brunkard et al. 2007; Siregar et al. 2015; Elaagip et al. 2020; Tellle et al. 2021). However, the communities that participated in the present research are far from urban centers (in average 31.5 km), which can lead to similar results in quantity of breeding sites presence, containing Culicidae individuals and infected people at that moment.