has wild habits with ecological plasticity in anthropized environments. We report the occurrence of this species in several types of artificial containers in 2 fragments of the Atlantic Forest in the city of São Paulo, Brazil. A total of 1,428 larvae were collected, in which plastic containers had the highest abundance of immature forms of Li. durhamii, with 579 (40.5%) specimens collected, followed by glass bottles with 464 (32.5%) and metal cans with 223 (15.6%). The high abundance of Li. durhamii in artificial containers points to an adaptation process to impacted environments. Therefore, studies that investigate the feeding habits and vectorial capacity of this species are essential to understanding its epidemiological role in the environment.
The genus Limatus Theobald belongs to the Sabethini tribe and comprises 9 species distributed in the Neotropical region with 5 species in Brazilian territory (Forattini 2002, Harbach 2007). Adults of Li. durhamii (Theobald, 1907) are active during the day and are found in the forests of South and Central America (Hervé et al. 1986). This species has been found naturally infected with several arboviruses, such as the Guama virus, Maguari virus, Tucunduba virus, and Caraparu virus (Segura and Castro 2007, Dias et al. 2022), all belonging to Orthobunyavirus genus (Peribunyaviridae family; Adams et al. 2017). Those viruses cause nonspecific symptoms, such as fever, headache, arthralgia, myalgia asthenia, and chills (Groseth et al. 2017, Matos et al. 2019).
Immature forms of Li. durhamii develop in canopy and soil breeding sites, such as broken bamboo, tree holes, fruit peels, escargots, stone holes (Lopes 1999, Hernández-Rodríguez et al. 2018), snail shells (Mangudo et al. 2017), and artificial containers (Lopes 1999, Zequi et al. 2005). Researchers observed Li. durhamii in the same artificial breeding sites with Aedes aegypti (L.), Aedes albopictus (Skuse), and Culex quinquefasciatus Say (Honório et al. 2006). The species were collected in artificial breeding sites in ecotopes with urban characteristics (Lopes 1999).
Immature forms (larvae and pupae) of the mosquito were collected monthly between March 2015 and April 2017 from artificial containers in 2 remnants of the Atlantic Forest: the Capivari-Monos Protection Area (CMPA) and the Cantareira State Park (CSP). Both sites are located in the metropolitan region of São Paulo state and serve to shelter, protect, and ensure conservation of biodiversity in eastern Brazil. Suction samplers were used for collection procedures. Three collection sites were selected at CMPA and CSP. Each site shows a different level of anthropization. The collected immature forms were placed in plastic containers with individual information and sent to the Laboratório de Entomologia em Saúde Pública, Faculdade de Saúde Pública, where they were monitored until the adults emerged and were euthanized with ethyl acetate. The dichotomous key by Lane (1953) was used for identification. Figure 1 shows an adult specimen and larvae of Li. durhamii.
Images of sampled specimens. (a) Limatus durhamii larvae photographed alive. (b) Li. durhamii adult specimen. (c, d) The arrows indicate the yellow metallic aspect of mesonotum scales: lateral and dorsal views (characteristic of this species).
Images of sampled specimens. (a) Limatus durhamii larvae photographed alive. (b) Li. durhamii adult specimen. (c, d) The arrows indicate the yellow metallic aspect of mesonotum scales: lateral and dorsal views (characteristic of this species).
A total of 1,428 Li. durhamii larvae were collected in artificial breeding sites (Table 1). The highest abundance of immature forms of this species was found in plastic containers, with 579 specimens collected, followed by glass bottles (464 specimens) and metal cans (223). Specimens of Li. durhamii were found in more types of artificial breeding sites in areas of CMPA than in the CSP. Some artificial containers e.g., water reservoirs, tarpaulins, tires, and asbestos roofs, were not found in CSP. Glass bottles were present only in the CSP environment. Plastic, metal, and ceramic containers were present in both sites. Even with less artificial container variability, CSP showed 28% more presence of Li. durhamii larvae than CMPA. It is worth noting that abundance may have been affected by the number of different kinds of containers scattered throughout the collection sites. The most abundant artificial containers were plastics (49 units), glass (21 units), and metal (20 units) containers.
Types of artificial containers in CMPA and CSP where Limatus durhamii specimens were collected.

The occurrence of Li. durhamii in artificial containers (tires, plastic, and metal containers) was previously reported (Lopes 1999, Zequi et al. 2005, Calderón-Arguedas et al. 2009, Ortega-Morales et al. 2010). Its tolerance to environmental pressures makes it the Sabethini species best adapted to artificial breeding sites (Forattini 2002). Our findings corroborate those of Lopes et al. (1993), who also found Li. durhamii in artificial containers such as plastic, tires, and metal, with the greatest abundance being in plastic ones. Calderón-Arguedas et al. (2009) also reported the presence of Li. durhamii in plastic and metal containers; however, it was more abundant in metal ones. Nonetheless, Lopes (1999) and Zequi et al. (2005) reported that Li. durhamii was the most abundant of all species in tires.
In addition, Li. durhamii larvae feed on decaying matter present in water, but in the absence of resources, they can be predate on larvae on other mosquito species as well (Lopes 1999). Lopes (1999) reported larvae of Li. durhamii coexisting in an artificial container with larvae of Culex eduardoi Casal and Garcia, Culex bigoti Bellardi, Cx. quinquefasciatus, and Toxorhynchites sp. in a riparian forest in an urban area in southern Brazil, and Marín et al. (2009) observed Li. durhamii and Ae. aegypti larvae living in the same artificial container. Honório et al. (2006) and Ortega-Morales et al. (2010) reported Li. durhamii preying on Ae. aegypti larvae, and Olano et al. (2015) registered Li. durhamii larvae coexisting with Ae. aegypti and Cx. quinquefasciatus within schools located in ecotypes with wild and periurban characteristics. The presence of Li. durhamii and Ae. aegypti larvae in the same artificial container (Marín et al. 2009, Olano et al. 2015) is indicative of its adaptation process to impacted and peridomestic environments.
Our scope does not measure water volume for each container; based on local observation, glass bottles, metal, ceramic, and plastic containers, and tires (more effective breeding sites) can accumulate less water volume than water reservoirs. Asbestos roofs and tarpaulins showed less frequency of Li. durhamii larvae due to the minimum potential water accumulation capacity, where leaves can occupy all of the spaces.
Although both localities were Environmental Protection Areas, each one has different conservation stages. This becomes evident when observing the different types of artificial breeding sites found. Plastic containers may be the best indicator of human disturbance. This artificial breeding site was the most common in the 2 localities. Tarpaulins, tires, and asbestos roofs were present only in the CMPA area. Glass bottles were found only in the CSP area. Unlike the CMPA, the CSP is the type of park where people use its trails as a place for leisure and nature appreciation. On these occasions, it is common to consume different types of drinks, and as a result, the bottles often are abandoned, becoming breeding sites for mosquito species.
The scant information about the biology of Li. durhamii contrasts with its medical-epidemiological importance (Lorenz et al. 2019). This species was found in high abundance in some culicid fauna investigations (Ceretti-Junior et al. 2020, Evangelista et al. 2021) and was registered as naturally infected with yellow fever virus (Obara et al. 2012). Barrio-Nuevo et al. (2020) detected Zika virus in Li. durhamii during a study in forest fragments of São Paulo municipality. Different from other Sabethini species, the high presence of Li. durhamii in artificial breeding sites may point to an adaptation process. The evidence of interaction of Li. durhamii and etiological agents shows the necessity of blood meal habit and vectorial capacity investigations of this mosquito species and its importance in public health scenarios.
We thank the State of São Paulo Research Foundation for providing financial support (FAPESP BIOTA Program 2014/50444-5; 2021/14677-9; 2023/11212-0; CAPES Process 88887.939180/2024-0) and the National Council for Scientific and Technological Development, Brazil (CNPq 301466/2015-7) for providing financial support. We are also grateful to Márcio Port de Carvalho, Vladimir Arrais and Aline Taminato at the Institute of Forestry for logistics support and for granting permission to work in the park and to the Institute of Forestry for hosting the researchers and granting a license for the project (SMA 260108-008.481/2014). We would also like to express our gratitude to the field and laboratory teams at the Department of Control of Endemic Diseases and the School of Public Health, University of São Paulo: Luis Filipe Mucci, Ana Maria Ribeiro de Castro Duarte, João Carlos do Nascimento, Paulo Frugoli dos Santos, Luis Milton Bonafé, Antônio Waldomiro de Oliveira, Daniel Pagotto Vendrami, Gabriela Cristina de Carvalho, and Amanda Alves Camargo.