**Molecular and Proteolytic Profiles of** *Trypanosoma cruzi* **Sylvatic Isolates from Rio de Janeiro-Brazil**

Suzete A. O. Gomes1,2 et al.\* *1Laboratório de Biologia de Insetos, GBG Universidade Federal Fluminense-UFF, Rio de Janeiro, RJ 2Laboratório de Transmissores de Leishmanioses Setor de Entomologia Médica e Forense IOC-FIOCRUZ-Rio de Janeiro, RJ Brazil* 

### **1. Introduction**

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Liu, S., - Liu, Y., - Huang, X., - Wang, H., - Niu, X., - Li, J., - Liang, L., - Luo, Y., - Ji, K., - Zhou, W., - Yu, Z., - Li, G., - Liu, Y., - Li, L., - Qiao, M., - Feng, L., - Zhang, K. (2011). Genomic and Proteomic Analyses of the Fungus *Arthrobotrys oligospora* Provide Insights into Nematode-Trap Formation. *PLoS Pathogens*, Vol. 7, e1002179, Chagas disease, also known as American trypanosomiasis, has its epidemiology conditioned to the (i) triatominae vectors, (ii) etiologic agent, *Trypanosoma cruzi*, and (iii) sylvatic and sinantropic reservoirs, the mammals. Social factors associated with economic factors, such as industry development, population growth and rural area colonization, which lead directly to ecological imbalance, provide favorable conditions for the disease establishment (Barretto, 1967; Ávila-Pires, 1976).

In 1909, Carlos Chagas releases his discovery on a new human disease, the American trypanosomiasis, subsequently known as Chagas disease. Carlos Chagas described the etiologic agent, the protozoan belonging to the Trypanosomatidae family *Trypanosoma cruzi*, and its insect vector belonging to the Hemiptera order, Triatominae subfamily, the so-called kissing bug (Chagas, 1909; Lent & Wygodzinsky, 1979).

The natural history of the Chagas disease probably started millions of years ago probably as a sylvatic enzooty, and it is still present in different areas from Brazilian territory. The arrival of men in these areas, as well as comprehensive deforestation caused by extensive farming during the past 300 years has caused triatomine insects, formerly sylvatic animal blood-sucking bugs, to meet men (Ferreira et al., 1996; Coura, 2007). Hence, the disease was characterized as a zoonosis, when men invaded the sylvatic habitat, deforesting and changing the ecological balance, and making triatomine bugs access to the residences.

<sup>\*</sup> Danielle Misael2, Cristina S. Silva2, Denise Feder1, Alice H. Ricardo-Silva2, André L. S. Santos3, Jacenir R. Santos-Mallet2 and Teresa Cristina M. Gonçalves2

*<sup>1</sup>Laboratório de Biologia de Insetos, GBG, Universidade Federal Fluminense-UFF, Rio de Janeiro, RJ, Brasil 2Laboratório de Transmissores de Leishmanioses, Setor de Entomologia Médica e Forense, IOC-FIOCRUZ-Rio de Janeiro, RJ, Brasil* 

*<sup>3</sup>Laboratório de Estudos Integrados em Bioquímica Microbiana, Instituto de Microbiologia Paulo de Góes (IMPG), Bloco E-subsolo, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, Brasil*

Molecular and Proteolytic Profiles of

establishing the disease sylvatic cycle (Coura & Dias, 2009).

*Trypanosoma cruzi* Sylvatic Isolates from Rio de Janeiro-Brazil 159

same ecotope he found infected *Triatoma geniculata* (*Panstrongylus geniculatus*) specimens,

Between 1913 and 1924 it became evident that the disease was not restricted to Brazil, being diagnosed in other countries in Central and South Americas, such as El Salvador, Venezuela, Peru and Argentina (Talice et al., 1940; Zeledón, 1981). In subsequent studies, Coura & Dias, 2009 mentions that Chagas (1924) demonstrated *T. cruzi* transmission cycle in the Amazon

In Rio de Janeiro state, the first Triatominae occurrence dated 1859, when Stal described *Conorhinus vitticeps* species, now called *Triatoma vitticeps*. At that time, Rio de Janeiro was

Following this finding, Neiva (1914) recorded the occurrence of *T. vitticeps* in Conceição de Macabu, formerly Macaé city district, presently Conceição de Macabu city. Due to information accuracy, Lent (1942) suggested it would be considered as the type location of *T. vitticeps*.

Subsequently, Pinto (1931, as cited in Lent, 1942) pointed out its presence in Magé, and Lent (1942) in Nova Friburgo, at Secretario location in Petrópolis city and at Federal District, which was Rio de Janeiro at that time. In Minas Gerais state, it was observed by the first

In Rio de Janeiro state other species were also found. Guimarães and Jansen (1943) collected *Panstrongylus megistus* specimens in a building by the hill, and identified *Trypanosoma cruzi* sylvatic reservoir (skunk), but did not find the sylvatic focus. Dias (1943) listed Chagas disease transmitters in Rio de Janeiro as being *Panstrongylus megistus*, *Panstrongylus geniculatus* (Latreille, 1811), *Triatoma vitticeps* (Stal, 1859), *Triatoma oswaldoi* (Neiva & Pinto, 1923), *Triatoma infestans* (Klug) and *Triatoma rubrofasciata* (De Geer, 1773), first recording the occurrence of *Schizotrypanum sp*-infected *P. megistus* in two districts in the capital of Republic (Santa Tereza and Botafogo). In 1953, in a survey performed at Araruama and Magé, Dias stated it was a relevant issue for the State, while Bustamante & Gusmão 1953 pointed out the presence of *T. infestans* at Resende and Itaverá cities. New findings have been identified, such as that of Coura et al. (1966), who found *P. megistus*, *Triatoma tibiamaculata* and *T. rubrofasciata* in three districts at Rio de Janeiro city, and that of Aragão & Souza (1971), who signalized the presence of *T. infestans* colonizing domiciles at two cities in Baixada Fluminense. In the same year, Coura et al. (1966) described some autochthonous instances of *T. infestans*-transmitted Chagas disease at Baixada Fluminense, and Becerra-Fuentes et al. (1971) recorded *T. rubrofasciata* occurrence at Morro do Telégrafo in the former Guanabara state. Silveira et al. (1982) performed an entomologic inquiry at Duque de Caxias and Nova Iguaçu cities (RJ), and only found *T. infestans* species. Ferreira et al. (1986) verified the occurrence of *T. vitticeps*, and positivity for *T. cruzi*-like forms, in 12 cities, of which the one with the highest incidence for both observations was Triunfo location at Santa Maria Madalena city. In 1989, a *P. geniculatus* specimen was found in a domicile at São Sebastião do Alto city (RJ) (personal communication with Teresa Cristina M. Gonçalves). The occurrence of *Rhodnius prolixus* (Stål, 1859) in Teresópolis was pointed out by Pinho et al. (1998), which caused questioning, once this species was restricted to the northern region of the country. Nowadays it is known this species does not occur in Brazil (Monteiro et al., 2000, 2003). *T. vitticeps* was found in Poço das Antas, Silva Jardim city, by Lisbôa et al. (1996), and in Santa Maria Madalena by Gonçalves et al. (1998). In both

time by Martins et al (1940), and in Espírito Santo state, as mentioned by Lent (1942).

region with the identification of this parasite in monkeys of *Saimiri scirius* species.

assigned as type location, without defining whether it referred to the city or state.

Therefore, the transmission cycle of *T. cruzi* is comprised by a sylvatic cycle, in which the parasite circulates among mammals and sylvatic vectors, and a domiciliary cycle, in which the infection is ensued by the contact of mammals, sylvatic vectors and sinantropic animals with domestic and domiciled animals, including men (Barretto, 1979).

Human Chagas disease, an antropozoonosis that evolved from a zoonosis, is strongly related with men's social class, type of work and habitation (Dias, 2000). During the 70's, the disease endemic area achieved at least 2,450 Brazilian cities, 771 of which were detected to have *Triatoma infestans*, the main disease vector in Brazil. At that time, there were over five million people affected by the disease in the country, with an incidence of approximately one hundred thousand new cases yearly and mortality above ten thousand deaths yearly. Less than five percent of blood banks used to control donors and over seven hundred cities had their homes infected by *T. infestans*. This situation led scientists to press the government to prioritize a national program against the disease. Homes from endemic areas were sprinkled with the appropriate insecticide and, in accordance with law; mandatory screening of blood donors was implemented throughout the country (Dias et al., 2002). The control program of the main vector in Brazil was recognized in 2006, with a certificate from the World Health Organization (WHO) for virtual elimination of *T. infestans* in Brazil (Dias, 2006). As the main vector was eliminated, currently there is a concern that other Triatominae species, formerly deemed secondary in the disease transmission, such as *Triatoma brasiliensis*, *Triatoma pseudomaculata* and *Panstrongylus megistus*, take the place of *T. infestans* in some locations, therefore becoming potential disease vectors in Brazil (Coura, 2009).

Despite the great progress in controlling vector and transfusion transmission in the countries from the Southern Cone, transmission is ongoing in other parts of the continent, and the issue of already infected people, most of whom are in the chronic phase of the disease, is still a challenge to public health (Urbina, 1999). Currently Chagas disease affects between twelve and fourteen million people in Latin America, and at least 60 million people live in areas with transmission risk (WHO, 2002). In Brazil, the disease notification became compulsory as per Ordinance V of Health Surveillance Secretary of Ministry of Health dated February 21, 2006.
