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Virus hunters

A team of researchers goes to the Amazon to watch for the entrance of emergent diseases

The July holidays involved a lot of work for a group of researchers and students from the University of São Paulo (USP). Aboard a jeep and a pick-up, they penetrated into the trails in the Amazon Rainforest on a mission to collect biological material from birds, insects and other animals. Their goal: to provide research material that sought to check up on the arrival of illnesses such as bird flu and West Nile fever, transported by migratory birds. The choice of the Amazon was not by chance. On the one hand, there is the richness of its biodiversity and the impact of uncontrolled deforestation that favors the appearance of emergent viruses. On the other, the Northern Region of Brazil is the entrance port of the main migratory routes of birds, which could bring these illnesses with them. “Epidemiological surveillance in the region is extremely important”, explains the biologist Luciano Matsumya Thomazelli, one of the expedition’s participants.

Once the job was done, during which they covered close to 9,000 km passing through ten Brazilian States, they returned to São Paulo with their bags full. They collected samples of blood, feces, oral secretions from more than 400 ducks, turkeys, wild birds, insects and bats, which are now being analyzed at USP’s Biomedical Sciences Institute (ICB). In this type of survey, the researchers search to find the first signs of the entrance of emergent diseases and, thus, deflagrate with rapidity prevention and treatment strategies. The first results of this effort are showing that, for now, there are no indications of infected animals. But the monitoring is going to continue and should be extended throughout various states. The threatening virus H5N1, which has produced epidemics in Asian countries, was detected in migratory birds found dead in various European countries.

The importance of the expedition, captained by professor Edison Luiz Durigon, a virologist from USP, can be understood from various parameters. In the first place, a mobile structure of monitoring, capable of coming from Sao Paulo and arriving by land to remote and poorly accessed locations was tested successfully. “This is a challenge that we have to meet with in the case of the emergence of some unknown disease. Now we’re prepared for this”, says Durigon. At any moment, they could take off again aboard the Land Rover jeep and the Ford pick-up with the capacity to transport ten people, two tents, one boat, laboratory equipment and an electrical generator. Such apparel is an appendix of a greater project. The idea is to collect material to feed the Klaus Eberhard Stewien Laboratory, established some two years ago at the ICB, the first of its kind in the country working to the NB3+ standard (the bio-security level of 3+). This is almost the maximum possible for civil research – there are more sophisticated installations only in developed countries.

In second place, the trip was useful for training future researchers. Among the eight members who journeyed with Durigon and a further four who made up the rearguard team, there were eight doctorate degree students, one master’s degree student and three undergraduates. During their journey, the majority had to deal with their first major field experience. “Never again will I look at a sample in a test tube in the same way as before. I learned to respect the work of whoever obtains the material”, says Jansen de Araújo, a biologist and doctorate degree student from ICB. The trip itself was important for reasons that would not themselves figure in academic curriculums. If the expedition assisted in the maturing of its researchers, it was also rich in adventure and in life experience situations. There were cinematographic landscapes (such as that of the bridge that collapsed seconds after the passage of their vehicles) and even nervous breakdowns, food rationing and at least one fall with injuries. This is without mentioning the curious personalities that the group ran into during the expedition’s 29 days of travel.

It was 1 p.m. on the 3rd of July when the expedition, made up of eight people, set off from  the parking lot at ICB, in University City, in the city of São Paulo. The departure crowned the super action efforts of the biologist and doctorate student Luiz Francisco Sanfilippo. Responsible for the field equipment, he shared with the undergraduate student Ricardo Lieutaud the task of mounting the expedition’s structure. A few days before departure, he discovered that the boat had been delivered without the required specifications  and he had to argue considerably with the supplier who ended up changing the merchandise. As well he become annoyed with the lateness in delivery of the roofing of the Ford pick-up. Biology student Lieutaud, invited to make the journey, could not go because of a family problem and was substituted by Miguel Augusto Golono, a biologist and doctorate degree student.

Four days later, after having spent the nights at Três Lagoas and Coxim , in the state of Mato Grosso do Sul, Cuiaba  and Comodoro, in the state of Mato Grosso, the group arrived at their first destination, the ICB Study Center in Monte Negro, in the of Rondonia, some 250 km from Porto Velho, the  capital of the state. They remained there for a week, collecting material and preparing themselves for what they would face ahead of them. Counting upon a well defined structure – the ICB Center has laboratories, sleeping quarters and communication equipment available -, they risked their first captures. They opened their nets at 4:30 a.m. to capture insects and birds.

During their stay at Monte Negro, the expedition gained its ninth member. The biologist Carolina da Silva Ferreira, a master’s degree student at ICB, convinced her colleagues to let her go along with them. Carolina had been in Monte Negro some two months, doing research that would be as a basis for her dissertation. Weary because of the long period away from home, one of her few fun things to do was to consume the inexpensive assai ice cream sold in Monte Negro – four balls for only R$ 2.50. ?In two months I?d already put on 10 kilos-, the biologist recalls. During the following weeks not only would she lose her accumulated weight but would be the leading person in some of the expedition’s most difficult situations.

The departure from Monte Negro, with destination Manaus city, took place at 11 a.m. on the 15th of July. The researchers planned to complete the 680-kilometer stretch in one day and told a group who would be waiting for them in Belem that, if in three days they had no news of them, then they should seek help. They spent the night in a hotel in Humaitá, a town in the state of Amazonas, just after the state border with Rondonia, and, on the morning of the 16th they faced the worst moment of their trip: the BR 319 highway, impassable during the rainy season, and a suffering even during the dry season and along which they would travel in trail jeeps. Without imagining what they would confront ahead, they made a major mistake ? they took little food. “They asked that I buy 20 rolls of bread and I bought 40, but it was little”, recalls the biologist Jansen de Araújo. The undergraduate biomedical student Tatiana Lopes Ometto, sums up what happened: “We left happy and enthusiastic, as we kept going on the road full of potholes,  our smiles faded. Some of us were worried. Others became highly nervous”.

Along the way they met up with a man on a motorcycle. He was a neighborhood inhabitant who received from the Amazonas government the mission of maintaining the wooden bridges along the BR 319. The man suggested that they go back. This was because the last bridge on the road had collapsed a few days earlier. “We went ahead even at that since the Land Rover and the pick-up could pass through small rivers”, says Durigon. The man recommended that we spend the night in a small village a kilometer or so further on.

The obstacles were many and the condition of the road was so precarious that at a certain point the group stopped to decide if we should even go one step further. Under the shade of the boat that the pickup was carrying they voted. The majority voted to continue with the journey – one of the few dissenting voices having been that of Luiz Francisco Sanfilippo, not by chance a member of the expedition who knew the region better. The tension reached the maximum level when a bridge collapsed seconds after the passage of their vehicles. Now there was no turning back. Time kept passing, the sun kept beating down – but nothing appeared of that village mentioned by the man on the motorcycle, or any other vestige of human life. The most stressed out with the situation was Miguel Golono. On the edge of a nervous breakdown, he demanded all of the three rolls of bread to which he had the right. He was calmed down by his colleagues. It was 11p.m. on that Saturday with a crescent moon when, debilitated, they decided to set up camp in the middle of the road, on a part of the pavement   that the forest and the rains had forgotten to swallow up.

Some, like Luiz Sanfilippo, slept, even outdoors – only much later would he know that wildcats inhabited the region. “That stop was paradise. I thought to myself, ‘we’re alive, let’s sleep'”, recalls Tatiana Ometto. The bread was consumed with apprehension – nobody could say how long they would still spend on that imitation of a roadway. They woke up  early and at 6 a.m. were already going along the road. New misfortunes, clearly, would be waiting. Carolina Ferreira, the biologist who had joined the group at Monte Negro, suffered an accident. She and her colleague Mario Luiz Figueiredo, a biologist and doctorate degree student, had the mission of analyzing the conditions of the bridges before the vehicles would pass over them. During one of these inspections, Carolina tumbled down the side and almost fell into the river. She spent the remainder of the journey with pains and scratches.

Two concerns marked this stretch of the journey. One was that bridge that no longer existed, according to the man on the motorcycle. The group was in for a good surprise. They found a brand new bridge, built hours before by a team that had been installing an optical cable between Manaus and Porto Velho. The other concern was their delay. Scheduled to take one day, this stretch of the journey had already lasted three – and they had no means of communication. They had been completely isolated. Cell phones didn’t work in that part of the forest. “I wouldn’t do the journey again without a satellite phone”, says  Durigon.

Some 5.5 kilometers from there, at Mosqueiro Island, in Pará, the support team made up of the biomedicine professional Danielle Leal de Oliveira and the biologists Juliana Rodrigues and Lílian Keller, was exasperated with the lack of news.  They had begun to discuss what to do and had checked out emergency telephones. At 11:00 p.m. on the 17th of July, the telephone at the house where they had been staying finally rang. They ran to answer to it. In vain. The line went dead. As the number had been registered they returned the call. At the other end someone answered a community telephone in Careiro Castanho, in Amazonas, some 88 km from Manaus. “It was a resident. I asked if there was a bearded man close by”, said Danielle, referring to the biotype of Edison Durigon. “They said that there was. Alas! They had managed it.”

Over the next two days the tension diminished as they carried out maintenance to the vehicles, went down by boat to Belém, and also prepared to visit the Pará capital – now by plane. They arrived at Mosqueiro island, some 79 kilometers from the Pará capital at 9:00 p.m. on the 19th of July. The goal was to visit duck breeders in the neighborhood of Belém. The support group formed by Danielle, Juliana and Lilian had already searched out addresses and visited breeders, from large breeding farms to those who bred them in their backyard, asking for authorization for the researchers, when they would arrive, to carry out the collection of biological material. They covered localities such as Marabitana, Vigia, Santa Bárbara, Santo Antônio do Taruá and Santa Isabel. Duck breeding is a long-standing tradition in the region – tucupi (a thick cassava sauce) duck  is typical food for lunch during Círio de Nazaré, Para’s religious feast day. “We went from house to house explaining to people. Some refused. The majority asked: will my duck die?”, recalls Juliana. They would not die, they emphasized. It would only be necessary to remove a little blood.

In these places it is common for the inhabitants to raise ducks, pigs, chickens and other domesticated animals all together, in the backyard of their house. And it is starting from this close excessive familiarity between man and animal that the contagion of illnesses such as Asian flu, a veterinary disease capable of infecting human beings, can evolve. The fear is that mutations of the Asian flu virus gain ways of transmitting themselves between human beings, thus producing a pandemic. “We took photos of these places and they look like the TV scenes from Vietnam, where there has already been a human outbreak of bird flu”, says ised professor Durigon. “The elements are the same: poor rural residences, surrounded by domestic animals. In one of these houses, the chickens laid their eggs in the kitchen.”

The state of Pará stay, which lasted a week, was productive. They collected material from wild birds, ducks and turkeys, and got to know places and people who they would never forget. “It was touching to see those simple people helping us. There was a man who went by bicycle to fetch a duck so that we could collect a sample”, says the veterinary student Renata Ferreira Hurtado. There they met a certain Ms. Maroquinha, a humble country woman who greeted the USP researchers with a banquet extracted from her backyard, with the right to duck and turkey well seasoned with coriander and annatto, garnished with rice, beans and sweet cassava well as cupuaçu cream for dessert. “They’re very poor, but nobody goes hungry”, says Durigon.

The return journey back to Sao Paulo, again aboard the Land Rover and the Ford pick-up, lasted four days, with stops at Imperatriz (PA), Paraíso do Tocantins (TO) and Goiânia (GO). Happy with the success of the enterprise, the researchers exhibited, above all, the scars of the adventure. Various had journeyed debilitated with gastroenteritis. Carolina Ferreira, she who had joined the group in Rondonia and who had injured herself on falling at the wooden bridge, spent the months of August and September resting. Soon after she arrived in São Paulo she discovered that she had contracted hepatitis A during her journey. It was in this manner that she lost the 10 kilos accumulated through the ice cream orgy during her stay in Monte Negro.