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Chapter II

Chagas disease in pre-Colombian civilizations

Felipe Guhl* and Arthur Aufderheide **

*Universidad de los Andes. Facultad de Ciencias. Departamento de Ciencias Biológicas. Centro de Investigaciones en Microbiología y Parasitología Tropical. Bogotá, Colombia.

** Department of Pathology, University of Minnesota, Duluth, USA

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KEY WORDS: Chagas disease, Fossil DNA, Pre-Colombian civilizations, human migrations, mummies.

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Summary

Human Chagas disease is a purely coincidental occurrence. As humans came into contact with the natural foci of infection, they might have become infected as a single addition to the already extensive host range of Trypanosoma cruzi, which includes other primates and a diversity of sylvatic mammals. Thus began a process of adaptation and domiciliation to human habitations, through which the vectors had direct access to abundant food as well as protection from climatic changes and predators. Historical evidence suggests that many pre Hispanic cultures were in close contact with triatomine insect vectors in their dwellings before the arrival of European conquerors to the New World, and several reports also provide evidence of the antiquity of human T. cruzi infection in South America, Central America and Mexico.Clinical manifestations of Chagas disease have been observed in Chilean and Peruvian -mummies; moreover, T. cruzi kinetoplast DNA (kDNA) has been recovered from Chilean mummies up to 9,000 years old.

Introduction

Reconstruction of the behaviour of a modern disease during antiquity is a formidable challenge. However, success in such an endeavour, would allow for the creation of a new database, and this new information could then spawn new hypotheses. Their results could then be blended with our present knowledge to produce an unbroken history of infectious diseases from deep antiquity to the present. Paleoecological integration of such data could help explain chronological changes, whose causes could be exploited for novel modern therapeutic or preventive control of the condition.However, there are currently only three methodological tools that can be used in such searches: genetic variation, archaeology and biochemistry.

Genetic Variation

More than 100 feral animal species in Central and South America can be septicemic with the trypanosome, but demonstrate few or no clinical effects of the parasite in their blood. Although this phenomenon could have occurred via several mechanisms, the most likely is host and/or vector genetic changes produced by a process of coevolution. For example, new antigens could have been added to the surface of the parasite, alternatively, or in addition, the parasite could have developed the ability to cross cell membranes and complete the remainder of its life cycle in an intracellular location where antibodies could not gain access to it (evasion). Such changes commonly occur over an extremely long time period, but result in an increased tolerance of both host and vector for each other. (Anderson and May, 1991; Schmunis, 1994).

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Archaeology

The modern domestic cycle is the common environment which humans are exposed to reduviid bugs, usually Triatoma infestans and Rhodnius prolixus. These insects hide in the cracks and defects of a house’s roof and wall during the day, emerging at night to obtain a blood meal from their prey. That prey includes not only humans, but also domestic animals. Thatched roofs are especially attractive for these insects (Edgcomb and Johnson, 1976). Even today, many families supplement their meals by raising guinea pigs in cages in the home as well as wild animals such as armadillos and small mammals, a high fraction of which can become infected by T. cruzi.One of the best-studied populations dealing with Chagas disease is from the coastal area of northern Chile at the foot of the western Andean slopes, between about 19 and 23 degrees of south latitude. The extremely arid climate here generates rapid, spontaneous desiccation of buried bodies, arresting the decay process. The absence of rainfall then preserves these dried bodies (mummies) for millennia.

Biochemistry (Bioarchaeology)

The parasite transmission cycle

T.cruzi was originally transmitted directly between marsupials, but wassubsequently vectored to other mammals through the advent of the blood-sucking Hemiptera (Triatominae) that are now considered the major vectors.This transfer from marsupials to other mammals may have been the main factor promoting adaptation of the parasite from the original widespread form (T.cruzi I) to a range of other lineages now grouped as T.cruzi II and T.cruzi III to T.cruzi VI. Current estimates suggest that the first divergence from T.cruzi I to T.cruzi II (mainly human), occurred about 10 million years ago (Mya), (Zingales et al.,2009) The epidemiological pattern of Trypanosoma cruzi reveals that primitive transmission was restricted to establishedcycles in tropical forest environments. Triatomine insects fed on small mammals in broad areas of the South American continent, with no human involvement in the natural cycle. The same situation persists today in the wild, where the disease maintains enzootic epidemiological character. The presence of T. cruzi does not seem to affect triatomines significantly, nor does it impact the mammals that have been naturally infected, suggesting that a balance exists between species as a result of long periods of adaptation and coevolution (Guhl et al. 1999 op cit.). Although in general the Hemiptera represent an ancient order, with fossilized remains dating from the Permian, nearly 232-280 Mya, it is possible that the triatomines evolved later starting at different times and from diverse ancestral forms.The Hemiptera comprise a large order with over 80,000 species widely distributed in all tropical and temperate areas. Ancestral predatory habits among the triatomines can be inferred from the fact that some species occupy a relatively wide spectrum of ecotopes and are able to exploit different species of hosts, while others, occupy restricted habitats

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and hosts (Schofield & Matthews, 1985). The vectorial transmission of T. cruzi is restricted to the New World.

Insect vectors associated with the human habitats

TThis also is true for Rhodnius prolixus, the primary domiciliated - vector in regions of Colombia, Venezuela and the vast majority of Central American countries. The development of R.prolixus from egg to adult takes 3 to 4 months whereas for species such as T. dimidiata, a peridomiciliary/domiciliary species, this may take 1 to 2 years.Triatomines show a high degree of dispersion, which involves two different mechanisms: one – passive, by the vertebrate host and the other active by walking or in the case of adult insects flying. Several authors have reported the passive transport of triatomines in clothing, baggage and transport and even the transport of eggs and nymphs in the feathers of birds (Foratinni et al., 1971, Gamboa et al., 1962)The geographic distribution of triatomine species extends from the Neo-tropics to Neoarctic regions and is closely related to environmental and ecological factors.

Map 1. Distribution of different triatomine species restricted to particular geo- ecoepidemiological factors

Historical data indicate that the disease was transmitted in South America and seriously affected the inhabitants of endemic regions, who referred the insects with vernacular names. The many indigenous names for the insect vectors such as vinchuca, hita and chirimacha demonstrate the frequency with which pre-Colombian civilizations encountered these insects. The Quechua word vinchuca, for example, means “bug that lets itself fall” which describes the behaviour of domiciliated insect after feeding on blood. Hita is also a Quechua word that means “bedbug” and the chirimacha meaning is “which fears the cold”. These Quechua words clearly evoke the domiciliated behaviour of triatomines.Quechua (“qheshwa”) is an indigenous language of the Andean region, spoken today by approximately 13 million people in Bolivia, Peru, Ecuador, Northern Chile, Argentina and Southern Colombia. It was the official language of Tawantinsuyu, the Inca Empire,which was the largest empire in pre-Colombian America. The administrative, political and military centre of the empire was located in Cusco in modern-day Peru. The Inca civilization arose in the highlands of Peru sometime in the early 13th century. From 1438 to 1533.

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Table 1. List the vernacular names for triatomine insects in Southamerican geographical areas and cultures.

(Adapted, modified and expanded, from Schofield and Galvao, 2009)

REGION NAMES MEANINGArgentina, Chile, Uruguay vinchuca1 bug that lets itself fallBelize bush chinche8 (implies absence of domestic Triatominae in

Belize )Bolivia vinchuca1 bug that lets itself fall

uluchi 1 bug without wings; refers to nymphal stagestimbucú

Brazil Barbeiro2 barber shaver

furão 2 big piercing bug

chupão 2 big sucking bugbicudo beaked bugfincão 2 big piercing bugcascudo 2 thick-skinned bug, used mainly for nymphs

chupança 2 sucking bug

procotó 2 bug that hides in cracks

Gigolô2 exploiter of women

percevejo 2 wall bedbug

gaudério 2 indigent thief

rondão 2 big bug that observes from hiding

percevejão 2 big bedbug

percevejo do sertão2 bedbug from the sertão,Sertão = interior of Brazil

percevejo das pedras2 bedbug amongst the stones

piolho de piassava 2 louse from the piassaba palm, refers to R.brethesi in Amazon region

vunvum 2 probably onomatopoeic for the sound of bug flightjosipak Matacos indians, Roraima

îipi Macuxi Indians ,Roraima and Venezuela,refersspecifically to T. maculata

Colombia Pito3 whistle or hornchupasangre 3 Blood sucker

kajta in kággaba4 Kogi Indians, refers to the spirit of the insect

kajta chiguibu4 the eggs of the triatomines

kajta bulo4 first nymph star

kajta yagua 4 second nimph star

kajta tema4 other star and adults

kajta ungaga4 the place for payment , where the spiritual Leader (Mamo) after consultation with the spirit of the triatomines , pays with offerings that vary greatly depending on the query in order to restore the natural balance

Cuba sangrejuela 3 BloodstealerEcuador Chinchorro3 large bugCentral America chinche besucona 3 kissing bug

talaje 3 cutting bug chuluyu needle polvoso 3 dusty

chinche bebe sangre 3 blood-drinking bug

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chinche picuda3 biting bug

Mexico

chinche besucona 3 kissing bug

chinche hosicona 3 trunked bug

chinche picuda 3 biting bug

chinchona 3 big bug

pech 5 onomatopoeic for the sound of bug flight

Texcan 6

Paraguay chichá guazú 7 big ug Itchajuponja7 bug suckersham bui tá 7 insect that does harm by its dejectionstimbucú 7 long beak

Peru chirimacha 1 bug that dislikes the coldYta 1

USA kissing bug 8 cone-nose bug, big bedbug

China bug 8 refers to T. protracta on Pacific coast, once assumed to come from the orient

red-banded cone-nose refers to T. rubrofasciata and /or T. sanguisuga

Venezuela chipo 3 little bugîipi Macuxi Indians; refers specifically to T. maculata

1Quechua, a native American language family spoken primarily in the Andes of South America, derived from an original common ancestor language, Proto-Quechua. It is the most widely spoken language family of the indigenous peoples of the Americas. 2Portuguese 3Spanish 4Kogui language, Colombia 5Maya language 6Azteca (Nahuatl) language 7 Guaraní 8English

Historical overview

Historical data allow us to infer that when the Europeans arrived to the New World, there was already a local knowledge of triatomine insects, including their habits and some biological characteristics directly related to man; however, these insects were not known to be associated with the disease first described by Carlos Chagas in 1909.Geronimo de Bibar, Chilean author of a chronicle from early 1500, writes " for six years now there is a type of bugs that sting very badly and give little itching, they are as big as cockroaches and their time is in summer". (Bibar,1966)Various chroniclers, such as Antonio de Ciudad Real, also make references to bugs with wings. There are other references to bugs with wings that correspond to Triatominae (i.e.,

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kissing bugs,pitos hyphae, hocicones) , (texcan in Nahuatl , pec in Maya and yta, in Quechua). Lizárraga also refers to this group of insects saying the following about Bolivia and Argentina:“ Here called hyphae ,cockroaches (black), as large as the insects found in the ships of the North Sea, and that colour, with wings: but with the difference that they are almost invisible, they sting and bite so delicately at night that it is not felt after the fire is finished, however after two days a welt rises as a bean, so itchy it is insufferable (...) The insects are afraid of fire, and when the fire goes out they fall down the walls or from the ceiling and bite the sleeper in the legs, in the head and face (...) They have clumsy feet, and when they have filled their bellies with the blood they have sucked, cannot walk.”Felix de Azara also describes these bugs “like flat beetles”, adding the important observation that “...when defecating on the wound, they leaved an indelible stain”.

When describing the existence “vinchucas” in Venezuela, the Jesuit José Gumilla, reports that the bite is painless, however, once the arthropod detaches, unbearable itching pain occurs. In Peru, especially in the area of Charcas, Bernabé Cobo comments that are harmful, locate their victims by scent and are known by the natives as hyphae (Gumilla, 1963).

Archaeological excavations have been carried out along the Aleutian Islands and the west coasts of Canada and the United States, as well as on the west coasts of Central and South America. The origin and spread of humans coincided with the latter part of the Pleistocene ice ages, and the geographical distribution of early humans was influenced by these ice ages in a number of ways. All of the excavations have yielded evidence of the presence of human activity since nearly 15,000 years ago. However, the precise date of the first human presence on the continent may be even earlier, as far back as 20,000 to 25,000 years, although this is still a matter of debate among archaeologists. For the purpose of this chapter, it is safe to conclude that humans were spreading south and east of North America and that these small bands of hunter gatherers had reached the northern tip of South America around 10,000 to 12,000 years ago. This amount of time was enough for the adaptation of various parasitic diseases to their new hosts. Cultural developments, such as agriculture and permanent or semi-permanent settlement patterns, created an ideal environment for the spread of infectious diseases, including tuberculosis and syphilis. Human cultural adaptation to warm and humid environments, allowed Chagas disease to spread widely.

Pre Hispanic settlements in areas of transmission of Trypanosoma cruzi The process of triatomine domiciliation occurred simultaneously with the process of human settlement near zoonotic transmission cycles of T.cruzi. The process was gradual, as has been demonstrated with other species of triatominae today.

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The insects found enough food sources, feeding on men and domestic animals such as dogs, chickens and guinea pigs. There is evidence that, as in modern times people living with pets in their homes in endemic areas enhanced the transmission of the parasite. By comparing the migration patterns of pre-Colombian cultures in different areas of Latin America with the distribution of triatomine species, it is possible to infer the degree of passive dispersion between the parasite/insect vector and pre-Hispanic settlements.One of the most extensive and detailed studies of this type was performed by Viana and Carpintero in 1977 and 1979, from which one can draw the following information relating to the major pre-Hispanic cultures in the Americas:

Argentine-Bolivian Altiplano. Northwest ArgentinaThis area received direct and indirect influences from the high Andean cultures and had a society based on an intensive agricultural economy, with a variety of vegetables, livestock and advanced bronze metallurgy. These cultures survived for a period of just over 1,700 years, which was grouped into three stages:Early Period: From the appearance of the earliest civilizations to the year 650 ADCultures: Tafi, Cienaga, Candelaria and Condorhuasi.Reviewing the detritus deposited in urns of Cultures Tafi, Santamaria and the Aguada, Remains of Triatoma infestans (which can easily recognized by an expert eye) were found in the detritus deposited in urns of Tafi, Santamaría and Aguada cultures, very easy to recognize for an expert eye.The sealing and subsequent burial of the urns would have prevented any triatomine penetration a posteriori. Clearly, the insects were buried at the same time, probably hidden in the clothes of the corpse, which obviously suggests a close contact between the vector and pre-Columbian aboriginal communities.Middle Period: From 650 to 850 AD. La Aguada CultureLate Period: From the 850 AD until about 1480 AD, beginning in the Inca period, ranging from the arrival of the Incas to the first entry of the Spanish conquerors. Cultures In the course of human settlements, natural pathways were formed, by which they conducted a lively exchange that enabled the economic complementarity of the different cultures. The most important archaeological sites (Quebrada de Humahuaca) show that the houses were important community nuclei and were built of stone walls and roofs of branches and mud, very good niches for triatomines.The economy was based on agricultural farming supplemented by the raising of Llamas ( Lama glama).

Sierras CentralesThis region, comprising the central region of Argentina-Sierras de Córdoba, San Luis, and Santiago del Estero, has been inhabited since the year 6000 BC. Interestingly the region of Santiago del Estero continues to have one of the highest prevalences of Chagas disease in Argentina and is one of most triatomine-infested areas.Sur del PerúFor the purposes of this chapter, we will highlight the Chilca culture (3 800 BC) and Nasca culture (2 500 BC). In Huanuco, Peru's Eastern Sierra, buildings were found that confirm the domestication of "cui" (Cavia sp.), dating 1,200 BC.

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In the Formative Period (1200 BC -100 AD), and continuing through the Regional Development Period (100-800 AD), the Wari Old Empire (800 -1,200 AD), and the Empire Tawantisuyo (1.430-1.532 AD), the human migration of these populations increased in northwest Argentina. The construction of large cities that still surprise us, agriculture and, the breeding of llamas and other mammals, were all good environments for domiciliated populations of T. infestans.New settlements and the displacement of entire villages, as a result of wars of conquest, facilitated the dispersion of triatomines into new areas.The mechanism of T. infestans adaptation to human habitats, probably facilitated by the custom of pet storage near or even inside houses. This custom still persists in many parts of Bolivia Perú and Ecuador, where the Guinea pigs are bred in homes for human consumption.These mechanisms of domiciliation helped T.infestans, became one of the first insects to adapt to human habitats in South America, and it remains probably the most widespread and numerous. Although its settlement of Uruguay and Brazil occurred more recently, (probably through Argentina), its dispersion increased, spreading northeast into Bahia (Brazil). T. infestans was one of the main vectors in the country (with Panstrongylus megistus), until a decade ago when efficient vector control were established. Paraguay could be settled from the Provinces of Northwest Argentina and across the Gran Chaco.

Map 3. shows the origin and dispersal of Triatoma infestans in South America.

Meso-America. Mayan Culture The great Mayan culture in the Peten region (northern Guatemala) lasted from 300 to 900 AD. However, the different communities that made up the amazing Mayan Empire, seems to have originated from what is now northern Honduras (Santa Rosa de Copan). In the Early Formative Period (1200 BC - 300 AD) the society was agricultural and built ceremonial buildings in stone. Constructions of this type also housed chiefs, priests and dignitaries, although most of the towns people occupied dwellings made of boughs and straw-reinforced adobe and poles, an environment conducive to the establishment of triatomines.After an intermediate period (900 - 1.000 AD) the New Empire or Mexican Period began, which took place primarily in Yucatan, Quintana Roo, Campeche, Tabasco and Chiapas (1000 to 1200 AD). The disintegration of the Empire began due to internal wars and was completed by the arrival of the Spanish (1,450-1,550 AD).The economy, mainly agricultural, reached a high level, and trade with the cultures of Central Mexico was frequent and important. There is also sufficient evidence of an active relationship, mostly commercial, with the northern cultures of South America.

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The keeping of poultry among the Toltecas, a Mayan village dominated by the Aztecs in the fourteenth century, certainly led to the adaptation of Triatoma dimidiata and subspecies to human habitats by a mechanism similar to that for T. infestans.Although its geographical distribution is wide, T. dimidiata is less efficient in transmitting the T.cruzi. It range extends north into Mexico, and south to Colombia and Ecuador, (Bargues et al., 2008)Interestingly, a culture akin to the Maya flourished in the area occupied by the current state of Oaxaca: the Zapotec culture. This area is inhabited by Triatoma barberi the Protracta complex species that is best adapted to human habitats. The adaptation of species and subspecies of the Phyllosoma complex which is found around dwellings and in domiciliary environments in southern and central Mexico seems to have been more recent occurred by a mechanism similar to that of the species mentioned so far.

Map 4 shows the origin and dispersal of Triatoma dimidiata in Mesoamerica and northern South America region.

Andean Region, Northern South AmericaThe Chibcha tribes, stretched from Colombia to Ecuador and Nicaragua. Their houses were built of posts or poles, with or without added adobe, and the roofs were built with palm leaves. The domiciliation process of Rhodnius prolixus from its wild habitat occurred as it does today. Genetic studies of these insects have shown identity of genotypes among domiciled insects and insects found in the wild, especially those nesting in palm trees of the genus Attalea. (Pintoet al., 2005).One of the most prominent Chibcha groups inhabiting the highlands of Bogota and its nearby valleys, were the Muiscas (men), who built several large communities (100 AD). They grew corn, potatoes, sweet potatoes, cotton and cassava in the lower valleys, and their extensive housing groups formed stable communities for the creation of a solid economy based on trade with neighbouring villages, primarily those to the north and west. Domiciliated R prolixus penetrated into Colombia and Venezuela and then expanded northward reaching Guatemala.

Map 5. Shows the dispersion of Rhodnius prolixus in the Americas.

Oral infection by T.cruzi

In addition to insect transmission, Chagas disease may be acquired by ingestion. Trypanosoma cruzi oral transmission is possible through food contamination by

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The faeces of the vector or by the ingestion of raw meat from infected sylvatic reservoirs. Many aboriginal cultures and rural populations in South America today still have the habit of eating raw or semi-raw meat from wild animals, which are reservoirs of the parasite.In the last 10 years, for example, many outbreakes of orally acquired acute Chagas disease have been reported in different geographical areas in South America. 152 cases, including 5 deaths and 121 acute cases in Brazilian Amazonia, 34 case including 4 deaths in Colombia, and a large urban outbreak of orally-acquired acute Chagas disease, at a school in Caracas, Venezuela that affected 103 schoolchildren (de Noya et al.,2010)In general, these outbreaks lead to death a percentage of infected individuals, which indicates a high pathogenicity of the parasites and demonstrates its capacity to penetrate through the gastric mucosa, despite the presence of gastric acid. (Hoft et al., 1996),(Marsden, 1967),( Maguire et al.,1986). Additionally the presence of metacyclic forms of T. cruzi in anal gland secretions of the opossum (Didelphis marsupialis), an animal with wild and peri-urban habits, cannot be overlooked as a source of oral transmission in the outbreaks.The traditional mechanism of transmission of Chagas disease, which involves contact with metacyclic forms in the faeces of wild triatomines ,may not be the most common type of transmission in wild ecotopes like the Amazon where several other types of transmission appear to be occurring.

Evidence of human Trypanosoma cruzi infection in pre Colombian civilizations

For more than a century, examination of skeletal tissue from ancient human remains has demonstrated information useful for the understanding of some diseases in antiquity (Aufderheide, 2003). Unfortunately, only a minority of human diseases leaves a detectable impact on bone. Hence, during the past few decades, efforts have been made to evaluate whether other diseases could be detected by examination of the soft (i.e., nonskeletal) tissues in mummified human remains.One of the first reports related to Chagas disease in human remains from South America was from Rothhammer et al. in 1985, which describes cardiac lesions compatible with the chronic clinical picture ofthe disease.The evidence was obtained from 35 bodies dating from 470 BC to 600 AD,that were mummified in the desert of Atacama in Chile.A Peruvian Inca mummy was studied by Fornaciari et al. in 1992, who showed evidence of Chagas disease in the lesions described; additionally they demonstrated the presence of amastigote nests in the heart muscle of the mummy.In 1997 and 1999, Guhl et al. op cit. initiated molecular studies of mummies from the Atacama desert. For the first time, they isolated T. cruzi DNA from 4000 years old mummified tissue.In 2000, Ferreira et al. published an article on mummies from San Pedro de Atacama,that confirmed chagasic infection in specimens up to 2000 years old.All of these reports confirm the hypothesis discussed in this chapter on human migration and Chagas disease.

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Most of these studies have taken the form of individual case reports. These are valuable and will remain so for a long time; however, the study reported herein represents an effort to determine whether examination of such mummified human soft tissues can reconstruct the behaviour of a disease in entire ancient populations. We selected American trypanosomiasis, more popularly known as Chagas disease, as an appropriate candidate because of its high prevalence in the area of our study. Initially, we attempted to detect the presence of Trypanosoma cruzi using molecular biology methods methods that targeted a segment of the parasite’s DNA in excess of 300 base pair. Although we succeeded in that effort (Guhl et al., 1999 op cit.), the target segment proved to be too long for the sensitivity needed for a study involving a large number of specimens. A shorter segment involving a probe had the necessary sensitivit, but required considerable manipulation of the amplified product (Madden et al., 2000, op cit.) Our final effort, described in detail in this report, used a short segment of kinetoplast DNA with less handling of our amplicon. This technique proved to have the sensitivity we needed with minimal manipulation to bring about the hybridization reaction. This was then applied to extracts of tissue specimens from 283 mummified human remains from a South Andean coastal zone. The results enabled us to construct the paleoepidemiology of Chagas disease in that area over a period of nine millenial, from the appearance of the first humans in that region to the near present (Auderheide et al., 2004, op cit.).Members of the Chinchorro cultural group were the first to settle this coastal segment. The oldest body from this group was radiocarbon-dated to about 9000 years ago. Stable isotope reconstruction of their diet indicates that approximately 90% of their diet was of marine origin, consistent with some of their grave artefacts. We have only a few samples of their housing because of lack of rain, which meant they did not need a waterproff shelter. Reconstruction of several of these shelters indicates that a series of slender wood poles were arranged in a circular pattern, and the top ends were gathered together in the form of a wooden tepee. The cover of the pole skeleton did not survive, but was most probably composed of intertwined reeds harvested from the brackish water at the river mouth. Such a dwelling would be ideal for nesting by the Chagas insect vector.After more than 5000 years of residence with this type of a marine-based strategy, the Chinchorros were replaced by highland migrants, that we call Alto Ramirez. Arriving at the coast in about 1000 B.C., they introduced agriculture. Their shelters were lightly larger than those of the Chinchorros, with walls of cane and reeds; however, because they still employed plant products in parts of their structures, these remained attractive for insect nesting.By 4000 B.C., the rapidly expanding highland population called Tihuanaco extended their territory to the sea, where it remained the dominant force until their empire crumbled about in 1000 A.D. Emerging from the resulting politico-cultural chaos of small, fragmented groups, were the people now known as Maitas Chiribaya. By this time, they had drifted from a mixture of marine and agricultural strategies more toward the latter, and this remained the case until the Incas arrived, preceding the Spanish.Beginning at the end of the Chinchorro period the trend in these societies was one of progressive technology development, particularly in textile production. Their shelters

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also slowly increased in sturdiness. However, local resources were limited, so opportunities remined available for insect vectors to move in permanently. The arrival of the Spanish initiated the historic period. The following cultural and societal chaos rarely offered the native populations improvements with respect to living quarters.Thus, the picture painted by archaeological findings of events in the past nine millennia begins with primitive dwelling structures that were altered by the succession of cultural groups until historic times, but not to a degree that would prevent infestation by the Chagas disease’s insect vector.Another interesting study of T. cruzi in human remains dating back 4,500-7,000 years were obtained from a Brazilian archeological site and. From these remains the recovery of an ancient DNA (aDNA) sequence corresponding to the parasite lineage I (T.cruzi I) was recovered.The mummy,was a woman ~ 35 years of age from a found –gatherer population. She was found in the Abrigo Malhador archeological site, Peruaçu Valley, Minas Gerais State. In this region, the semiarid ecosystem is predominant, thus it has a dry climate,and soil with a basic pH. These conditions have contributed to preservation of specimens.In this report, (Lima et al., 2008) showed that T. cruzi human infection in Brazil is ancient, dating back at least 4,500 years and therefore occurring in hunter-gatherer populations largely preceding T. infestans domiciliation. The presence of the T. cruzi I in humans 4,500-7,000 years ago in Minas Gerais State, where this genotype is currently absent, suggests that the distribution pattern of T. cruzi genotypes in humans has changed over time. Moreover, the recovery of an aDNA sequence and the possibility of genotyping parasites from human remains make it possible to reconstruct the early dispersion patterns of T. cruzi subpopulations. On the basis of these results, one may speculate that the current outbreaks of human T. cruzi infection, independent of triatomine domiciliation, are re-emergences of the ancient epidemiologic scenario of Chagas disease in Brazil. Paleoparasitological studies of Chagas disease may clarify the antiquity of this disease in the Americas through extraction and amplification of T. cruzi aDNA from human remains, other animal hosts and vectors fragments found in archaeological findings. Phylogenetic analysis of this material would also shed light on different aspects of host-parasite coevolution and parasite transmission cycles.

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