News Article | April 20, 2016
When the medical charity Médecins Sans Frontières called the worldwide shortage of snake antivenom a public-health crisis last September, Brazilian biochemist Paulo Lee Ho wasn’t surprised. He has spent his career at São Paulo’s Butantan Institute searching for better ways to create antivenom to treat bites from coral snakes. Conventional methods rely on natural coral-snake venom, which is hard to come by: the snakes produce only small amounts with each bite and are hard to raise in captivity. So Ho and others have turned to proteomics and synthetic biology in the hope of improving the quality and availability of antivenom. “We need a new way to meet the demand for antivenom from the Ministry of Health,” he says. These efforts are now bearing fruit. Last month, Ho and his colleagues reported1 that they had engineered short pieces of DNA that, when injected into mice, triggered antibodies against coral-snake venom. The scientists then boosted the animals’ immune response by injecting them with small pieces of synthetic venom antibodies synthesized in Escherichia coli bacteria. In a separate study also published last month2, another group of researchers in Brazil used synthetic antibody fragments to neutralize the effects of bites by the pit viper Bothrops jararacussu. Such progress is encouraging, given the severe medical burden caused by snakebites in the developing world, says Robert Harrison, head of the Alistair Reid Venom Research Unit at the Liverpool School of Tropical Medicine, UK. Each year, around 90,000 people die after being bitten by venomous snakes3. Yet antivenoms are still made using a method that has not changed for more than a century. Large animals, typically horses, are injected with small amounts of purified proteins extracted from snake venom, which prompts the production of antibodies. Plasma containing these antibodies is then given to snakebite victims. But this life-saving treatment is limited in important ways. Each antivenom is effective against only a single species or, at most, a small group. And the drugs must be refrigerated, a major problem in tropical countries without reliable electricity. “When you think about it, it’s amazing these antivenoms work at all,” says Leslie Boyer, director of the Venom, Immunochemistry, Pharmacology and Emergency Response Institute at the University of Arizona in Tucson. The number of pharmaceutical companies that make antivenoms is declining, because the drugs are not very profitable. In 2010, for instance, pharmaceutical giant Sanofi of Paris, ended production of the antivenom Fav-Afrique, which is designed to treat the bites of ten of Africa’s most poisonous snakes. Ho hopes that his approach will help to fill this void. Rather than relying on venom milked from live coral snakes, he began with small pieces of coral-snake DNA that code for venom toxins. He and his colleagues injected these DNA pieces into mice to prime their immune systems; a month later, they gave the animals a booster shot containing synthetic venom antibodies. Only 60% of mice injected with a lethal dose of coral-snake venom survived after receiving Ho’s experimental treatment, compared to nearly 100% for existing antivenoms. But Ho is undaunted. “This result shows there are other ways to obtain neutralizing antibodies,” he says. “Maybe to get better results, we need to try again but use more antibodies. We just don’t know yet.” The second Brazilian team, led by molecular biologist Carla Fernandes of the Fundação Oswaldo Cruz (Fiocruz) biomedical research institute in Porto Velho, tested a different technique, using a phage display library to make synthetic versions of the antibodies that llamas produced when they were injected with B. jararacussu snake venom. Giving these antibodies to snakebite victims would eliminate the need to use animal plasma. It also could reduce muscle damage and tissue death at the site of the bite, compared to traditional antivenoms, because the synthetic antibodies are smaller and better able to penetrate into tissue. The path to newer antivenoms isn't straight, but researchers believe that moving quickly is key. “There has been significant, rapid progress in this area, but it needs to be fast. There are too many people dying from what is essentially a preventable disease,” says Harrison. To Boyer, however, the antivenom shortage is not caused by a lack of science. “It costs 14 bucks to make a vial of antivenom that costs US$14,000 in the US,” she says. “You’re not going to get cheaper than that. The expensive parts aren’t the science — it’s everyone wanting a cut of the profits that drives the price up and puts it out of reach.”
News Article | February 28, 2017
SÃO PAULO, BRAZIL—A promising dengue vaccine faces an uncertain future in Brazil, scientists say, after the dismissal of a prominent immunologist who has been overseeing clinical trials of the preparation here. Last week, the São Paulo state government removed Jorge Kalil as director of the Butantan Institute, following accusations of administrative wrongdoing leveled against him by a former colleague. Announcing Kalil’s dismissal on 21 February, Governor Geraldo Alckmin praised him as a “great scientist” and said he wished he would continue to lead Butantan’s dengue vaccine program. Kalil told Insider he will decline that invitation. He denies the accusations against him and says it’s impossible to continue leading the vaccine program from outside the institute. “This is not an isolated project; it’s something that requires a coordinated effort by the entire institution,” he says. “I can’t accept something like this.” Scientists here and abroad have protested Kalil’s dismissal. Butantan researchers and staff staged protests last week, and the institute’s influenza vaccine factory shut down for a half day on Friday, demanding his return. In a 23 February letter to Governor Alckmin, Anna Durbin of the Johns Hopkins Bloomberg School of Public Health in Baltimore, Maryland, who led the initial clinical trials of the vaccine, wrote that the dengue vaccine program made “tremendous progress” under Kalil, and that this momentum may be “reversed by the removal of his leadership of the Butantan Institute.” The U.S. National Institute of Allergy and Infectious Diseases (NIAID) developed the early version of the TV003 vaccine—designed to protect against all four serotypes of the dengue virus—and sponsored the initial clinical trials, which were completed in 2012. NIAID then licensed the preparation to the Butantan Institute, a state-owned biomedical research and vaccine production facility here, to further develop the vaccine and conduct clinical trials in Brazil, where dengue fever is an endemic threat. The Butantan-produced vaccine is now being tested at 14 institutions across the country, in a $100 million, phase III randomized trial sponsored by Brazil’s federal government. Stephen Whitehead, a senior associate scientist at NIAID who led development of the vaccine’s attenuated viruses, also wrote to Alckmin, arguing that Kalil’s leadership skills were “essential” to the program’s continuation and that his removal from Butantan could have “dire public health consequences for the State of São Paulo and Brazil.” “His ability to solicit support for sound scientific endeavors, develop the framework for their realization, and reach out for international partners is indispensable for the success and credibility of the institute,” Whitehead wrote. Brazilian scientific organizations have also lobbied on Kalil’s behalf. In a public letter to Alckmin released on 22 February, the Brazilian Society for the Advancement of Science and the Brazilian and São Paulo academies of sciences ask that “apparent political conflicts of interest involved in this crisis do not interrupt projects that are so relevant to Brazil and the world”. In statements to the press, the São Paulo State Health Secretariat stated that Kalil was fired because of “grave administrative problems” detected at the institute during this tenure—he has been director since 2011—and for not respecting hierarchy. Kalil believes his removal resulted from personal and political motivations. He says he will refocus now on his other appointments at the University of São Paulo’s Medical School and Heart Institute, where he is a senior professor of immunology. Alckmin has appointed Dimas Tadeu Covas, a professor of hematology at the University of São Paulo’s Ribeirão Preto Medical School, as Butantan’s new director. “I do hope the person selected to lead the dengue vaccine program understands the immense effort it takes to see a vaccine through,” Durbin wrote in an email to Insider. *Update, 3 March, 2:39 p.m.: In a 2 March statement, the Health Secretariat of the São Paulo state government declared that “the development of the tetravalent dengue vaccine and other research projects of the [Butantan] Institute will not be affected by the exit of professor Jorge Kalil.” It further stated that “Butantan will remain strong as an international center of excellence in science, culture and public health.
News Article | March 17, 2016
Forget mosquito bites. Volunteers let researchers inject them with the dengue virus in the name of science - and an experimental vaccine protected them. Next up, scientists plan to use this same strategy against dengue's cousin, the Zika virus. It's called a human challenge, a little-known but increasing type of research where healthy people agree to be deliberately infected in the quest for new or improved vaccines against a variety of health threats, from flu to malaria. Wednesday's dengue study offered more evidence that what sounds bizarre not only can be done safely, it can offer important clues for how well a shot might work. "What we're trying to do is accelerate vaccine research," said senior author Dr. Anna Durbin of Johns Hopkins University's school of public health. It may be the best way "to know if you have a stinker before you try to test it in thousands or tens of thousands of people." The dengue candidate proved highly promising, researchers reported in the journal Science Translational Medicine. Dengue fever may have slipped from the headlines as the related Zika virus sweeps through Latin America, but every year mosquito-borne dengue causes devastating outbreaks throughout the tropics and subtropics. While most people survive dengue with few or even no symptoms, more than 2 million a year suffer serious illness and about 25,000 die. Creating a vaccine has been tough. It must work against four separate strains of dengue, and a shot that's only partially protective might backfire. That's because people who survive one type of dengue can suffer worse symptoms if they're later infected with another strain. Enter an experimental vaccine created at the National Institutes of Health, made from four live but weakened dengue strains. Initial studies had suggested the shots were safe and promising. But, "we really wanted to have an early clue that it was go to work," especially against the hard-to-prevent dengue serotype 2, said Dr. Stephen Whitehead of NIH's National Institute of Allergy and Infectious Diseases, who led the vaccine development. Researchers at Hopkins and the University of Vermont gave 41 healthy people who'd never been exposed to dengue either a single dose of the vaccine or a dummy shot. Six months later, those volunteers were challenged - injected with a weakened version of that dengue-2 strain. The results were striking: All 21 people who'd gotten the real vaccine were completely protected - while all 20 who'd gotten a placebo had dengue virus in their bloodstream and either a mild rash or a temporary drop in white blood cell count, researchers reported Wednesday. This kind of study mimics "the closest that it can be to what may happen in natural infection," said Dr. Nikos Vasilakis, a virologist at the University of Texas Medical Branch in Galveston, who wasn't involved in the new work but calls the NIH shot "one of the better vaccine candidates." Based in part on the findings, the Butantan Institute in Brazil last month began recruiting 17,000 people, ages 2 to 59, for the final testing needed to prove how well the NIH vaccine works against dengue in real-world conditions, when it is spread by mosquitoes. A competing vaccine, made by Sanofi Pasteur, recently was approved by Brazilian regulators for ages 9 to 45. What about Zika, the dengue relative that's been linked to babies born with unusually small heads? Already, researchers are planning similar challenge studies that could start even before there's a vaccine candidate, Durbin said. "We see a Zika challenge model as really beneficial for not only vaccine development but also to learn more about Zika itself," she explained. "We know very little about Zika right now," including how long it stays in blood and other parts of the body. Key to these challenge studies: Scientists must modify a virus strain in the laboratory so that it doesn't make volunteers openly ill but still is strong enough to spark a mild infection, what Whitehead called "that perfect in-between." Plus, that mimics what happens with both dengue and Zika, where most people who become infected never report symptoms. Before deliberately infecting someone, "you have to know that it's a completely controllable situation, that it's a mild and controlled infection," said Dr. Beth Kirkpatrick, who directs the University of Vermont Vaccine Testing Center that tested the dengue model.
Demasi M.,Butantan Institute |
Laurindo F.R.M.,University of Sao Paulo
Cardiovascular Research | Year: 2012
Vascular smooth muscle cell (VSMC) plasticity implies a capacity for rapid change and adaptability through processes requiring protein turnover. The ubiquitinproteasome system (UPS) is at the core of protein turnover as the main pathway for the degradation of proteins related to cell-cycle regulation, signalling, apoptosis, and differentiation. This review briefly addresses some structural UPS aspects under the perspective of VSMC (patho)biology. The UPS loss-of-function promotes direct cell effects and many indirect effects related to the adaptation to apoptosis/survival signalling, oxidative stress, and endoplasmic reticulum stress. The UPS regulates redox homeostasis and is redox-regulated. Also, the UPS closely interacts with endoplasmic reticulum (ER) homeostasis as the effector of un/misfolded protein degradation, and ER stress is strongly involved in atherosclerosis. Inhibition of cell cycle-controlling ubiquitin ligases or the proteasome reduces VSMC proliferation and prevents modulation of their synthetic phenotype. Proteasome inhibition also strongly promotes VSMC apoptosis and reduces neointima. In atherosclerosis models, proteasome inhibitors display vasculoprotective effects and reduce inflammation. However, worsening of atherosclerosis or vascular dysfunction has also been reported. Proteasome inhibitors sensitize VSMC to increased ER stress-mediated cell death and suppress unfolded protein response signalling. Taken together, these observations show that the UPS has powerful effects in the control of VSMC phenotype and survival signalling. However, more profound knowledge of mechanisms is needed in order to render the UPS an operational therapeutic target. © 2012 The Author.
News Article | February 3, 2016
LONDON (Reuters) - The world is once again asking scientists and drugmakers to come up rapidly with a vaccine for a viral disease that, in the latest case, few people had heard of until a few weeks ago, and even fewer feared. Making a shot to generate an immune response against Zika virus, which is sweeping through the Americas, shouldn't be too hard in theory. However, producing a safe, effective and deliverable product to protect women and girls who are at risk is not easy in practice. For a start, scientists around the world know even less about Zika than they did about the Ebola virus that caused an unprecedented epidemic in West Africa last year. Ebola, due to its deadly power, was the subject of bioterrorism research, giving at least a base for speeding up vaccine work. This time, the knowledge gap is more daunting. There are just 30 mentions of Zika in patents, against 1,043 for Ebola and 2,551 for dengue fever, according to Thomson Reuters Derwent World Patents Index. And there have been only 108 high-profile academic papers on Zika since 2001, against more than 4,000 on Ebola, as found in the Web of Science. Still, the U.S. National Institutes of Health, the Public Health Agency of Canada and the Butantan Institute in Brazil have started work on potential candidates for a Zika vaccine, and several biotech firms are in the race. They include NewLink Genetics, which helped develop the first successful Ebola vaccine with Merck & Co. Importantly, there is now a "big gun" vaccine maker with skin in the game: Sanofi said on Tuesday it will launch a Zika vaccine programme, a day after the World Health Organization declared the disease and its suspected links to birth defects an international health emergency. Japan's Takeda Pharmaceutical also said on Wednesday it had created a team to investigate how it might help make a vaccine, while GlaxoSmithKline is concluding feasibility studies to see if its vaccine technology could be suitable. Canadian researcher Gary Kobinger told Reuters he believes an experimental Zika shot might be able to be used on a limited emergency basis as soon as late 2016, although full regulatory approval will take years. Ben Neuman, an expert on viruses at Britain's University of Reading, says there are many hurdles ahead. "To be useful, a Zika vaccine would need to be effective and safe, but it's difficult to do both," he told Reuters. "It's a balancing act." That's because a good vaccine works by provoking the immune system into a strong response - but not enough to make a person sick - and there is no simple way to assess the right immune response for Zika, according to one drug company expert. Zika infection is so mild in the vast majority of cases that its victims are unaware they are even infected, so this group of potential patients is unlikely to need or want immunisation. The crucial target group is women who may be pregnant, since the disease's greatest suspected threat is the possible link to severe birth defects. All of this makes developing and testing a vaccine highly complex, especially since pregnant women are often excluded from clinical trials until the safety of new drugs or vaccines is well-established in other population groups. "It raises special safety considerations in vaccine development because you want to make sure any vaccine is safe for both mother and child," Takeda's vaccine head Rajeev Venkayya told Reuters. It also makes for an uncertain and potentially limited market for any Zika vaccine. Assuming Sanofi or another company succeeds in developing one, the vaccine may be used only in teenage girls - protecting them before they are likely to become pregnant - in countries and regions where Zika-carrying mosquitoes thrive. "It's a public health good initiative, it's not necessarily a commercial initiative," said Berenberg Bank analyst Alistair Campbell. "Zika is something that has cropped up suddenly and may well dissipate, so there may not be a sustainable annual cohort of patients for vaccination." Still, the WHO and other public health authorities will be relieved that one of the world's top drugmakers, Sanofi, has pledged to work on a vaccine and other big players may soon join the effort.
News Article | February 2, 2016
LONDON (Reuters) - The world is once again asking scientists and drugmakers to come up rapidly with a vaccine for a viral disease that, in the latest case, few people had heard of until a few weeks ago, and even fewer feared. Making a shot to generate an immune response against Zika virus, which is sweeping through the Americas, shouldn't be too hard in theory. However, producing a safe, effective and deliverable product to protect women and girls who are at risk is not easy in practice. For a start, scientists around the world know even less about Zika than they did about the Ebola virus that caused an unprecedented epidemic in West Africa last year. Ebola, due to its deadly power, was the subject of bioterrorism research, giving at least a base for speeding up vaccine work. This time, the knowledge gap is more daunting. There are just 30 mentions of Zika in patents, against 1,043 for Ebola and 2,551 for dengue fever, according to Thomson Reuters Derwent World Patents Index. And there have been only 108 high-profile academic papers on Zika since 2001, against more than 4,000 on Ebola, as found in the Web of Science. Still, the U.S. National Institutes of Health, the Public Health Agency of Canada and the Butantan Institute in Brazil have started work on potential candidates for a Zika vaccine, and several biotech firms are in the race. They include NewLink Genetics , which helped develop the first successful Ebola vaccine with Merck & Co . Importantly, there is now a "big gun" vaccine maker with skin in the game: Sanofi said on Tuesday it will launch a Zika vaccine programme, a day after the World Health Organization declared the disease and its suspected links to birth defects an international health emergency. Canadian researcher Gary Kobinger told Reuters he believes an experimental Zika shot might be able to be used on a limited emergency basis as soon as late 2016, although full regulatory approval will take years. Ben Neuman, an expert on viruses at Britain's University of Reading, says there are many hurdles ahead. "To be useful, a Zika vaccine would need to be effective and safe, but it's difficult to do both," he told Reuters. "It's a balancing act." That's because a good vaccine works by provoking the immune system into a strong response - but not enough to make a person sick - and there is no simple way to assess the right immune response for Zika, according to one drug company expert. Zika infection is so mild in the vast majority of cases that its victims are unaware they are even infected, so this group of potential patients is unlikely to need or want immunisation. The crucial target group is women who may be pregnant, since the disease's greatest suspected threat is the possible link to severe birth defects. All of this makes developing and testing a vaccine highly complex, especially since pregnant women are often excluded from clinical trials until the safety of new drugs or vaccines is well-established in other population groups. It also makes for an uncertain and potentially limited market for any Zika vaccine. Assuming Sanofi or another company succeeds in developing one, the vaccine may be used only in teenage girls - protecting them before they are likely to become pregnant - in countries and regions where Zika-carrying mosquitoes thrive. "It's a public health good initiative, it's not necessarily a commercial initiative," said Berenberg Bank analyst Alistair Campbell. "Zika is something that has cropped up suddenly and may well dissipate, so there may not be a sustainable annual cohort of patients for vaccination." Still, the WHO and other public health authorities will be relieved that one of the world's top drugmakers has pledged to work on a vaccine. GlaxoSmithKline is also investigating Zika and a spokeswoman reiterated on Tuesday it is concluding feasibility studies to see if its vaccine technology might be suitable. Ultimately, developing vaccines is a question of priorities, as evidenced by a patchy pattern of protection against a range of mosquito-borne viruses over the past 80 years. There was early success with the development in 1938 of the first vaccine against yellow fever, which belongs to the same virus family as Zika. More recently, drugmakers have successfully developed shots against Japanese encephalitis and dengue.
Kerkis I.,Butantan Institute |
Caplan A.I.,Case Western Reserve University
Tissue Engineering - Part B: Reviews | Year: 2012
Dental pulp from deciduous (baby) teeth, which are discarded after exfoliation, represents an advantageous source of young stem cells. Herein, we discuss the methods of deciduous teeth stem cell (DTSC) isolation and cultivation. We show that based on these methods, at least three different stem cell populations can be identified: a population similar to bone marrow-derived mesenchymal stem cells, an epithelial stem-like cells, and/or a mixed population composed of both cell types. We analyzed the embryonic origin and stem cell niche of DTSCs with respect to the advantages they can provide for their future use in cell therapies and regenerative medicine. In vitro and in vivo differentiation of the DTSC populations, their developmental potential, immunological compatibility, tissue engineering, and transplantation use in studies in animal models are also the focus of the current report. We briefly describe the derivation of induced pluripotent stem (iPS) cells from DTSCs, which can be obtained more easily and efficiently in comparison with human fibroblasts. These iPS cells represent an interesting model for the investigation of pediatric diseases and disorders. The importance of DTSC banking is also discussed. © 2012, Mary Ann Liebert, Inc.
News Article | November 25, 2015
A court in the Brazilian state of São Paulo has cut off distribution of a compound that is hailed by some as a miracle cancer cure — even though it has never been formally tested in humans. On 11 November, to the relief of many cancer researchers, a state court overturned earlier court orders that had obliged the nation’s largest university to provide the compound to hundreds of people with terminal cancer. Although the reversal applies only to requests for the drug by residents of São Paulo state, administrators at the university estimate that it covers about 80% of the orders they have received for the compound. The compound, phosphoethanolamine, has been shown to kill tumour cells only in lab dishes and in mice (A. K. Ferreira et al. Anticancer Res. 32, 95–104; 2012). Drugs that seem promising in lab and animal studies have a notoriously high failure rate in human trials. Despite this, some chemists at the University of São Paulo’s campus in São Carlos have manufactured the compound for years and distributed it to people with cancer. A few of those patients have claimed remarkable recoveries, perpetuating the compound’s reputation as a miracle cure. Dismayed by this unofficial distribution of phosphoethanolamine, the university’s administration moved in September 2015 to shut it down. Patients took the university to court, and in October 2015, Brazil’s Supreme Federal Court ruled in favour of one plaintiff who wanted the right to try the compound. A lower court then began granting orders for the university to provide it to others. University officials say that they were soon overwhelmed by more than 800 requests. “The decision not only ignored the opinion of medical specialists, but also overlooked the fact that the drug has only been tested on animals,” says bioethicist Volnei Garrafa at the University of Brasilia. “Such court decisions bring false expectations for patients and their families, creating turmoil in society and confusion between what is safe and what is not.” The Brazilian constitution guarantees universal access to health care, and it is common in Brazil for patients to turn to the courts to access drugs that the state health-care system does not dispense because of their cost, says Garrafa. But phosphoethanolamine presents a different situation, he adds, because it is not really a ‘drug’ at all. It is not approved by Brazil’s National Health Surveillance Agency. Those who argue that people who are terminally ill have a right to try experimental medicines saw the decision earlier this year as a significant victory. But to the university administration, drug regulators and cancer researchers, it showed blatant disregard for the basic scientific principle that a drug should be demonstrated to be safe and effective before being given to patients outside of a clinical trial. “It’s a violation of the autonomy of the university,” says Marco Antonio Zago, a physician and president of the University of São Paulo. “We are seen as a factory to produce something that we do not believe should be done.” Phosphoethanolamine is an important building block of the lipids that make up cell membranes. The compound can also act as a molecular signal that activates certain cellular processes. Although some studies do suggest that the compound may kill cancer cells in isolated cells and mice, it is not entirely clear how the compound brings about this response. Biochemist Durvanei Augusto Maria at the Butantan Institute in São Paulo believes that the compound may be imported into tumour cells and, once inside, trigger processes that cause the cell to self-destruct. Immunologist James Venturini at São Paulo State University and his colleagues have found that phosphoethanolamine may modulate the immune system’s response to cancer or affect cell division (M. S. P. de Arruda et al. Braz. Arch. Biol. Technol. 54, 1203–1210; 2011). But to justify using phosphoethanolamine in people, Venturini says, one would have to rigorously test it in a series of clinical trials using human volunteers. “I strongly believe that double-blind, randomized clinical studies are necessary,” he says. And even before such trials, further preclinical studies would have to be done, says Jailson Bittencourt de Andrade, secretary for research-and-development policy at Brazil’s science and technology ministry. The ministry plans to fund those studies, he says, and has already asked several research laboratories in the country to do the work. If those tests and subsequent clinical trials are successful, he says, the ministry will also fund the research needed to scale up phosphoethanolamine production to the quantities and quality needed for an approved drug. That process will take years. In the meantime, lawyers representing people with cancer have vowed to appeal against the latest ruling. If those appeals succeed, de Andrade worries that people will not wait until all the tests are completed, and may even abandon conventional treatment in favour of phosphoethanolamine. “Many patients have come forward and said they have tried the drug and it has worked for them,” he says. “So the other patients and their families — they want phosphoethanolamine now.”
News Article | March 4, 2016
Coral snake venom carries significant neurotoxicity and human injuries can be severe or even lethal. Despite this, antivenom treatments are scarce due to challenges collecting adequate amounts of venom needed to produce anti-elapidic serum. PLOS Neglected Tropical Diseases highlights exciting new research from the Butantan Institute in Brazil using synthetically designed DNA to produce coral-snake antivenom.
News Article | March 4, 2016
A pair of elongated, whiplike legs that are actually sophisticated environment sensors distinguish an unusual arachnid known as the whip spider, also called the tailless whip scorpion. Scientists recently described eight new species of this long-legged spider that are native to Brazil, nearly doubling the number of known species in the genus Charinus. Whip spiders use only six of their eight legs for walking, reserving their "whips" — which can reach several times the spiders' body length — for exploring the world around them and locating prey, through a combination of touch and chemical signals. Thanks to the new species discoveries, Brazil now boasts the greatest diversity of whip spiders in the world. But the forest ecosystems where these new species live are threatened by human development, and the researchers suggested that stronger conservation measures are urgently required in order to protect the whip spiders' habitats, and to discover more species before their habitats are destroyed. [Ghoulish Photos: Creepy, Freaky Creatures That Are (Mostly) Harmless] There are 170 known species of whip spiders found all over the world, mostly in tropical areas in the Americas. According to the researchers, the Amazon region — known for its diverse habitats, plants and animals — was long suspected of hiding many more whip spider species than were previously known. Though some whip spiders measure up to 10 inches (25 centimeters) at the fullest extension of their "whips," most are less than 2 inches (5 cm) and are hard to spot, hiding in leaf litter, under stones and tree bark, and in caves. To identify the new species, the researchers turned their attention to specimens from the collections in four Brazilian natural history museum collections: the Butantan Institute, the National Museum of Brazil, the Museu Paraense Emi?lio Goeldi, and the Museum of Zoology of the University of São Paulo. What does it take to describe a new whip spider species? Days, weeks and ultimately months of scrutinizing the spiders' body parts under a microscope and comparing them with other known species in order to find unique and differentiating characteristics, said study co-author Gustavo Silva de Miranda. De Miranda, a graduate student at the Center for Macroecology, Evolution and Climate at the University of Copenhagen, told Live Science that he and his colleagues performed exhaustive inventory of the spiders' features, including the number of segments in the whiplike limbs, the prey-catching spines at the tips of their legs, the groupings of their eyes, and the shape of the females' genitalia, called gonopods. "If we compare all these things and see that it's very unique, then we consider it a new species," de Miranda said. Genital structures turned out to be quite an important point of comparison, de Miranda explained. In each whip spider species, the female's gonopod shape corresponded very specifically to the shape of the male's sperm sac, for perfect alignment. But even as new whip spider species are described, their behavior and habits in the wild remain elusive, de Miranda said. One study, he said, detailed confrontations between males competing for females or territory — the spiders extend and display their head appendages, squaring off without actually fighting, and the loser (the one with the smaller display) retreats after a 20-minute stare-down. "But there is still a lot to be discovered," de Miranda said. "We're trying to understand the evolution of the group, their relationships, how they are so widespread, their morphological evolution." He said this makes it imperative not only to find new species, but to preserve the fragile ecosystems where these spiders live. "If they are not protected, they will vanish from nature," de Miranda said. The findings were published online today (Feb. 17) in the journal PLOS ONE. Follow Mindy Weisberger on Twitterand Google+. Follow us @livescience, Facebook & Google+. Original article on Live Science. Copyright 2016 LiveScience, a Purch company. All rights reserved. This material may not be published, broadcast, rewritten or redistributed.