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Venoms in the forest

Biodiversity makes Brazil a partner that international cataloguing initiatives seek

INSTITUTO BUTANTAN /ANTONIO COR DA COSTAInhabitant of the Amazon Region: rattlesnake (Crotalus durissus)INSTITUTO BUTANTAN /ANTONIO COR DA COSTA

In December, Hipócrates Chalkidis started going frequently to the National Forest of Tapajós, near the city of Santarém in the state of Pará, along the banks of the Tapajós river, one of the Amazon’s largest tributaries. Chalkidis and a group of biology students from FIT/Faculdades Integradas do Tapajós college are planning to bury dozens of buckets to collect snakes over the course of a year and a half. Moreover, the scorpions and spiders that fall into the traps will not be used merely to expand knowledge about the region’s biological wealth. Denise Cândido, a biologist from the Butantan Institute, delivered to Chalkidis, in December, a portable venom-extracting device, built by electrical engineering students and professors from PUC-SP, the Catholic University in São Paulo. The venoms will fuel research into new toxins, one of the lines of a broad program of research that for the last four years has brought together the experts from the Butantan Institute and from the scientific and medical centers in Pará.

With a bit of luck, Chalkidis and his team will collect several samples of the black scorpion of Pará, Tityus obscurus. This insect, which is entirely black and as much as nine centimeters long, is the cause of most of the 1,300 annual cases of scorpion bites reported in the North of Brazil. With more animals at hand, the Butantan team will be able to work faster to solve a mystery: the serum produced by this institute in São Paulo against the venom of the Tityus serrulatus does not appear to neutralize the effect of the venom of the black scorpion of Santarém on the nervous system, although it does seem to be effective against the neurotoxic action of the black scorpion of  Belém, the Pará state capital. “They might be different species, even though morphologically they are identical,”  ponders biologist Antonio Brescovit, from Butantan. At UFPA (the Federal University of Pará) in Belém, Pedro Pardal studies the genetics of scorpions to learn what actually differentiates them.

Widespread skills
In an article published in 2003 in the tropical medicine journal Revista da Sociedade Brasileira de Medicina Tropical, Pardal showed that accidents consisting of scorpion bites in Santarém had unique features, with mainly neurological symptoms – probably because, according to the studies of Lourival Possani, a Brazilian researcher who works in Mexico, one of the approximately 60 toxins of the venom, Tc1, is very small and therefore perhaps capable of passing through the barriers that protect the brain. Black scorpion bites cause strong muscle contractions (i.e., spasms), along with tachycardia and hypertension, besides making speech difficult. At the Santarém municipal hospital, physicians Mariana Quiroga and Paulo Abati found that Diazepam, used to fight anxiety and convulsions, may help to control the spasms of victims of black scorpion bites. “It was the only means we found,”  she says, arguing that the spasms brought on by the black scorpion’s bite are similar to the neurological symptoms of severe tetanus, which can be treated with Diazepam.

The team, which comprises Butantan’s most experienced researchers, is identifying, gathering and mobilizing experts previously dispersed over several research centers in Pará, such as Chalkidis, Pardal and Mariana, or Rosa Mourão, who heads a UFPA group in Santarém that has found chemical compounds able to stop the hemorrhage brought on by snakes’ venom. These were identified in the extracts of 18 plants from the region that the inhabitants employ normally. “The locals of indigenous descent take a syrup made from anti-ophidian plants before going into the jungle,” she says. “Alternatively, they apply a macerated plant on the bite to get rid of the pain or diminish inflammation.”  According to her, the vegetable extracts may contain enzyme inhibitors, such as phospholipases and proteases that, if duly researched, may lead to new anti-ophidian drugs or fight other diseases underscored by strong inflammation processes, such as arthritis. “It wouldn’t make sense to do anything in parallel, without making use of local skills,” commented Otávio Mercadante, Butantan director, while opening the fourth annual meeting that described the progress and the plans of the teams from the two states at an auditorium at FIT, in late October. “Butantan is never going to replace or compete with the local institutions. Our work will be of a complementary nature.”  Ever since he started to visit Pará in his search for suitable places in which to conduct research, Mercadante formed alliances with four higher education institutions (the State University of Pará, the Federal University of Pará, the Tapajós college and Iespes, the Instituto Esperança college), besides the Museu Paraense Emilio Goeldi museum in Belém and PSA/Projeto Saúde and Alegria, a social organization that aids riverside communities.

Appeased resistance
“This is a major learning opportunity,”  noted Mercadante, who also got the support of city councils, the state government, the research finance foundations of the states of Pará and São Paulo, the Ministry of Education and the Ministry of Science and Technology, in order to put three lines of research in motion: Amazonian biodiversity, the action of animal toxins, and the health history of the region. Complementarily, work is being conducted with local healthcare agents and physicians with regard to treating accidents involving poisonous animal bites. He soon found out that he would have to tread carefully. “We cannot get to the communities without the mediation of local culture, or we might be regarded as invaders.”  The collaborations reduce fears of encountering resistance among the São Paulo researchers. “Thousands of theses are written involving the knowledge and flavors of the region but they never get back to the area,”  recalled Magnólio de Oliveira, the deputy coordinator of PSA, “but now we have a sound team with a winning spirit.”

For three days, 80 students, along with medicine and nursing professionals, attended a course on accidents caused by venomous animals, such as snakes, spiders, scorpions, caterpillars and rays. In Santarém, 37 students and biologists followed another course on the diversity of poisonous animals in the Amazon Region. This included practical activities in the neighboring town of  Belterra. Giuseppe Puorto, a researcher and director of the Butantan Biological Museum, headed the presentations to professors, students, healthcare agents, community leaders and firefighters of Santarém. With the team and the Abaré boat, from the Saúde and Alegria project, he visited riverside communities on the Tapajós river and talked in a relaxed manner as he took out stuffed animals from his rucksack. He listened to descriptions of accidents with poisonous animals and, while acknowledging the home remedies, recommended to the inhabitants that they should never tie up or bite wounds. The Butantan team has already developed and distributed a booklet on poisonous animals, but there is still a lot to do.

Butantan has kept in contact with animals from the Amazon Rainforest for quite some time. This region remained isolated from the others until the early twentieth century, due to communication and transport difficulties. Nevertheless, back in 1914, according to a survey by Maria de Fátima Furtado and Myriam Calleffo published in Cadernos de História da Ciência, Emília Snethlage, then the director of the Goeldi Museum, sent a collection of serpents from Pará to Butantan for identification and storage. The transfer of animals has never ceased and today the institute in São Paulo has 6,625 samples from 213 Amazon Region sites. In 1924, Vital Brazil Mineiro da Campanha, the institute’s very first director who later resumed this position, hired the physician Jean Vellard to help to identify poisonous snakes. Vellard worked with Vital Brazil on the serum against the venom of the grass spider Lycosa raptorial, studied the toxicity of other snakes, identified new species and conducted many expeditions to the region to collect animals.

Eduardo CesarBelterra: forest, wooden houses and Ford’s heritage. Below, Cajueiro School: poisonous animals game, based on photos of local animalsEduardo Cesar

More recently, about 20 years ago, Brescovit travelled through the region’s forests, also in pursuit of Amazonian spiders, at a time when environmental changes in the region were still few. The trip from Belém to Santarém could only be undertaken by boat and took one week (today, it takes less than two hours by airplane). The physician Fan Hui Wen, along with several colleagues, has been visiting riverside communities for the last 10 years, to show how to avoid and treat poisonous animal bites without using a tourniquet, which can worsen the wound but is still used a lot in the region. Mariana Quiroga tells us that she recently treated a man who arrived at the hospital with a tourniquet made out of the snake that had bitten him.

Butantan is now emphasizing joint activities. “We want the study of these poisons to result in training researchers from Santarém, who will then return to the region to work there,”  says Ana Moura, a Butantan researcher and professor of graduate courses on the natural resources of the Amazon region at UFPA in Santarém. Up to now, two people have come south, José Pedro Marinho de Souza and Andria de Paula Santos da Silva, both of them graduates of FIT. After spending one year at Butantan in São Paulo, they returned to Santarém to collect animals and are expected to come back south in March, to complete their specialization course at Butantan in São Paulo. “There are a lot of brilliant people around, but it’s very expensive to leave Pará,”  tells us Marta Fernandes, a recent biology graduate who for one year monitored people bitten by scorpions, who had been treated at the Santarém municipal hospital. “I felt as if I were at a hospital in wartime,”  she says. Marta rode a bicycle in her quest for people and the animals; she managed to reconstitute the details of 45 accidents. “It’s a passionate matter to do research here.”

Ana Moura is integrating the Pará teams with the other ones from INCTTox (the National Science and Technology Institute on Toxins), headquartered at Butantan. “The collaboration between the teams can make it easier to research drugs based on animal and plant toxins, making use of the investments already made in buildings and equipment,” she says. “For the collaborations to progress,” says Osvaldo Augusto Sant’Anna, INCT coordinator, “it is fundamental for the researchers from Pará to be aware of  how science is conducted in São Paulo. INCE wants to generate knowledge jointly, rather than merely transfer technical knowhow.”

The people from São Paulo are being well received. “The Butantan knowledge is coming to us with the aim of empowering people,” stated Geraldo Pastana de Oliveira, the mayor of Belterra, a 12-thousand inhabitant town 48 kilometers away from Santarem, the nearest city, with almost 300 thousand inhabitants. The size of each municipality is not the only contrast. Santarém is centered around its port, one of the most important in the North, and the riverside walkway that vanishes into the horizon along the Tapajos, whose blue-green waters mix at that point with the muddy waters of the Amazon. Belterra is less explicit. Whoever arrives there in the early afternoon may get the feeling of having reached an abandoned town of wooden houses, reminiscent of early twentieth century movies. There is no one to be seen, as obviously everyone takes a siesta to get some respite from the intense and humid Amazonian heat.

There is now little dust on the long, straight main streets, which were paved with tarmac a few years ago, but that continue to be lined with hydrants, another peculiar reminder of this town’s history. Belterra was one of two towns founded in the twentieth century by the American entrepreneur Henry Ford to produce natural rubber from the rubber tree (see Pesquisa FAPESP, issue 158, April 2009). The other such town was Fordlândia, 130 kilometers from here, whose plantations with almost two million rubber trees soon came to grief as a result of a plant disease. The 3.2 million rubber trees of Belterra fared better, growing on more fertile land, and they survived this plague. For decades the town produced a lot of rubber, in a routine that was interrupted, from time to time, by fires -hence the hydrants scattered everywhere.

Chardival Moura Pantoja was born in Belterra 70 years ago and, as he says, grew up in “Henry Ford’s daycare centers,”  studied at the schools built by Ford and worked in the rubber plantations and in the production of natural rubber. He experienced prosperous times, when the richer inhabitants went to the golf course and those less well off, to the cinema. Soon after World War II, the demise of the enterprise started, triggered by the appearance of synthetic rubber, which was cheaper than the natural kind. This was followed by the rubber plantations being abandoned. The town’s hospital, which previously served the entire region, caught fire and never entirely recovered.

A different view
Pantoja was a federal civil servant in the late 1970’s, during harder times. He left the area and spent some years in other states. He chose to return and for 10 years he led a battle for the emancipation of the town, won in 1997. “We didn’t want to be subordinate to Santarém,”  he argues. Now he and other inhabitants, who get together at the end of the afternoon to chat on the benches that stand outside the homes, monitor with satisfaction the movements of the Butantan team, apparently eager to take part in another grandiose adventure. “Right from the start [the Butantan researchers] looked for me and were considerate enough to keep me posted about everything they were doing,” Pantoja told us. “They are trying to integrate into the society and to help solve our problems.”  Butantan is to set up its advanced research base in the Amazon Region in Belterra, on a 64-hectare plot of land that is still woodlands. “My dream is to also have a science lab there, to serve the students and teachers in the primary education network,” says Mercadante. “It’s perfectly feasible.”


A multidisciplinary team that includes the physician Fan Hui Wen is reconstructing the history of healthcare in Belterra. In collaboration with Maria Amélia Mascarenhas Dantes, from the University of São Paulo (USP), the group has recorded and filmed long chats with people such as Pantoja, who helped to build the town. Inhabitants such as Edineusa Medeiros Alves, also known as Neusa and who owns a pharmacy, and Arlison José Santos Reis, nicknamed Lica, who has a guesthouse, call Hui “Doctor”,  as if this were a simpler synonym for her name. She treats everyone with respect, as if each one had a PhD and a vast academic curriculum. What really matters is passing the inhabitants? Xray-like inspection, accepting the silences in conversations, and being a good listener. “Some animals that are dangerous as far as we are concerned are not dangerous to their mind. The boa constrictor, they say, is not normally poisonous; for some reason, they say, it’s only poisonous in August,”  says Hui. “One must get the other view.”

Around there, to treat ray bites, the inhabitants pour hot water or cast puffs of tobacco smoke on the wound. “It makes sense,” states Francisco Siqueira França, a Butantan physician, “because the venom is sensitive to high temperatures.”  Most people from Pará take plant-based home remedies to avoid or to treat snake bites. Magnólio, who usually visits riverside villages in the boats of the Saúde and Alegria project, tells us that he has already listed about 200 home remedies. One of them, called Pau-X, is special. Ana Moura found that this tea made from roots inhibits hemorrhagines, the enzymes in the snake venom that cause hemorrhaging, and these inhibitors might have other medical applications. The problem is that the formulation of Pau-X is secret and is safeguarded by the religious tradition of the region’s shamans. “Who am I, a biochemist, compared to a shaman?”  she asks herself, facing an impasse for which so far there has been no way out.

The Project
Sub-program of action in the Amazon Region (nº 08/57898-0); Coordinator Ana Moura da Silva – Butantan Institute; Investment R$345,000.00 (INCT-Tox/FAPESP)