Latest Update on How to Make Snake Antivenom in India
India needs to look for effective lifesaving Anti-Snake Venoms from Latin America
India is the home of nearly 275 snake species, of which 60 are considered venomous and medically relevant, and with various levels of toxicity.
SOURCE- https://www.financialexpress.com/lifestyle/health/india-needs-to-look-for-effective-lifesaving-anti-snake-venoms-from-latin-america/2224651/
Snakebite in India has remained a much-neglected subject in spite of the significant mortality and morbidity it causes.
By Sandeep Wasnik,
India is the home of nearly 275 snake species, of which 60 are considered venomous and medically relevant, and with various levels of toxicity. Out of these 60 venomous species, only four venomous snake species are responsible for causing the greatest number of medically significant human snake bite cases in India. These four venomous snakes are collectively referred to “Big Four” — Common Krait (Bungarus caeruleus), Russell’s viper (Daboia Russelii), Indian saw-scaled viper (Echis Carinatus), and Indian Cobra (Naja Naja).
In India, every year between 45,000 to 50,000 people die because of snake bites and others are left with amputations and permanent disabilities. This accounts for half of all the snake-related deaths in the world, where WHO estimates that 81,000–138,000 people die each year from snakebites worldwide. About 94 per cent of snakebite deaths occurred in rural areas, and 77 per cent occurred out of hospital or due to the unavailability of specialised hospitals. In the world, snakebite every month caused 10,417 total deaths, whereas compared to death due to EBOLA in 26 months is 11,325. This clearly indicates that death caused by snakebite is more than 2000 per cent as compared to death by EBOLA (436 deaths per month), this seems serious subject for the countries, the national and International Health Organisations.
Snakebite in India has remained a much-neglected subject in spite of the significant mortality and morbidity it causes. If diagnosis and treatment is given in a timely fashion the victim can go back to a productive life. The World Health Organisation (WHO) has called to half global snakebite deaths by 2030 and this will require substantial progress in India. The WHO also says complications linked to attacks make the phenomenon one of the most neglected of tropical diseases (NTD).
India’s Anti-Snake Venoms and its effectiveness.
In 1895, the first anti-venom was developed by Albert Calmette against the Indian Cobra. There is only one type of “Anti-Snake Venoms'' (ASV) used for any snakebite treatment in India, this ASV is polyvalent snake antivenom, a mixture made from venom extracted from the ‘Big Four’ snakes. But this ASV can be ineffective for those victims bitten by snakes outside of the Big Four. For production of Anti-venom, only horses are used and the venom utilized is a combination of those collected from the ‘Big Four’ species. Although this treatment has saved many lives, it is weakly effective as snake venoms and their toxins vary significantly across all subspecies and only 10 to 15 per cent of the antibodies in the sera bind to the venom.
The snakebite treatment in India is a kind of `Hit’ and `Trial’ as well as very expensive, if one vial of antivenom did not work, then 20 vials can be injected into the snakebite victim. The geography of India is extremely diverse and toxicity of snakes also depends on their geography, to understand in a simple way, each species and sub-species of snakes of different regions displays variation in the protein composition of venom.
India has to learn about life saving model of anti-venom from Latin America
Snakebite is a huge problem in Central and South America, there are the jungles, the Amazon and the rain forests, but in this part of the world, affordable Anti-Snake Venoms (ASV) are produced by numerous public institutions along with strong commitments by Government Health Ministries. Snakebite deaths and disabilities have been kept in check in Latin America. The production of anti-venom in Latin America can be considered a model for India and other parts of the world. In Latin America, there are 137,000 to 150,000 snakebite reports and 3400 to 5000 deaths per year. Several poly-specific and few mono-specific anti-venoms are manufactured in the region, with two laboratories (one public and one private) in Argentina, Colombia, and Mexico, four public laboratories in Brazil, and one public laboratory in Bolivia, Peru, Venezuela, and Costa Rica.
In Latin America, the network of public anti-venom manufacturing laboratories has established an informal dynamics of inter-group consultations on specific aspects, which has become a useful mechanism for troubleshooting in antivenom production. Technology transfer projects and cooperation contribute to the upgrading of anti-venom manufacture in public laboratories.
Venoms and Antivenoms of Brazil
According to the herpetologist of Instituto Butantan, Marcelo Ribeiro Duarte, Brazil has about 412 species of snakes, being 15 per cent poisonous. In this country, the following snake bites anti-venoms are produced: for rattlesnakes (Genus Crotalus), pit-vipers (Bothrops, Bothrocophias, and Lachesis), and Coral snakes (Micrurus and Leptomicrurus)".
In the Americas, the first public institution in charge of manufacturing anti-venoms was Instituto Butantan(English: Butantan Institute), created in 1901 in São Paulo, Brazil, under the leadership of Vital Brazil Mineiro da Campanha, after that in 1907 the Fundação Ezequiel Dias “FUNED” (English: Ezequiel Dias Foundation) was created in Minas Gerais and then in 1919,Vital Brazil moved to Niteroi, State of Rio de Janeiro to establish antivenom production at InstitutoVital Brazil. The production of anti-venoms at a national level coordinated between the Ministry of Health and the four manufacturing laboratories, Instituto Butantan, FUNED, Instituto Vital Brazil, and the Centro de Pesquisa e Produção em Imunobiológicos “CPPI” (English: Center for Research and Production of Immunobiologicals).
Brazilian Snake Island: Nearly 33 kilometers away from the coast of São Pauloin the Atlantic Ocean, yet no one lives there, in fact the Brazilian Navy forbids any visitors at all and with good reason, it's probably the most dangerous place on the earth but as deadly as they are the golden lance head Viper czar, golden lancehead one of the most venomous snakes in the world, actually save lives because their venom is being used to produce new blockbuster medications.
The island is also known as Snake Island, the island is small in size, only 43 hectares (106 acres), and has a temperate climate. Island is the home of estimated 3000 specimens of snake and in which 2000 golden lancehead, around one snake every300 meters, has the highest concentration of venomous snakes in the world.
Venoms and Antivenoms of Costa Rica
The Instituto Clodomiro Picado “ICP” (English: Clodomiro Picado Institute), established in 1970, attached to the Faculty of Microbiology of the Universidad de Costa Rica, responsible for the production of snake anti-ophidic serums and scientific research on serpents and their venoms and is one of the world’s leading manufacturers of snake antivenoms, and the only one in Central America. Costa Rica is home to 23 species of venomous snake, including the Central American “Bushmaster—one of the world’s largest vipers, growing up to 11 feet—and the “Bocaracá”, whose indigenous name means “Devil” that brings death when it bites. The production of anti-venom is produced by injecting the snake venom in horse and then drawing the horses’ blood, separating the venom-resistant antibodies, and mixing them into a fluid. Instituto Clodomiro Picado also produces anti-venoms in much the same way but with more advanced processes allowing for a purer product.
Today, Instituto Clodomiro Picado is not only producing the Anti-venoms for the country but for also for the LatAm regions as well as for the African and Asian countries at affordable prices. For example, Sri Lanka, a US based NGO (Animal Venom Research International “AVRI”), is supporting the development by Instituto Clodomiro Picado (of the University of Costa Rica) of a pilot batch of anti-venom against the venoms of the most dangerous Sri Lankan snakes.
The venoms were collected from Sri Lankan snakes to develop the Anti Snake Venom in Costa Rica. Once this anti-venom has been developed and pre-clinically tested, a clinical trial will be undertaken in Sri Lanka. If the new anti-venom is demonstrated to be clinically effective and safe, a technology transfer agreement among AVRI, Instituto Clodomiro Picado and the Sri Lankan authorities is expected to establish sustainable local anti-venom production.
How Brazil and Costa Rica could help to develop Anti-Snake Venoms for India?
Similar arrangements involving experienced anti-venom manufacturers and technology developers could be promoted in India. Successful international collaborations in the biopharmaceutical field, involving the so-called innovative developing country, would encourage and provide useful guides for such projects. Exporting the diverse snake venoms (according to geography) of India to Brazil and Costa for anti-venom development and clinical trial will be undertaken in India.
The involvement of World Health Organisation, other International health organizations, Ministry of health, Ministry of External Affairs in efforts to produce safe, effective anti-venoms locally or regionally is of paramount importance also promote awareness and help coordinate regional and global efforts in this area through regional training workshops and other activities to promote guidelines on anti-venom manufacture and control. Also, programs can involve between India-Brazil-Costa Rica for workshops and seminars, as well as a variety of activities aimed at enhancing the anti-venom production, quality control and capacity of manufacturers.
Now an anti-venom for local snake species
IISc ties up with Bharat Serums to make India’s first test batches of region specific anti-venom. Existing products are only applicable for the Big Four snake species— the spectacled cobra, common krait, Russell’s viper, and saw-scaled viper. ‘The Sind krait extends all the way from Punjab to Karnataka. Considering the toxicity of this snake and death resulting from it, we need ASVs that are effective against them,’ says Dr Kartik Sunagar, head of study
For the first-time in India, a government body, namely the Indian Institute of Science (IISc), Bengaluru, is producing test batches of non-commercial anti-snake venoms (ASVs) to evaluate the effectiveness of regional anti-venoms in neutralising toxins in region-specific snake bite cases compared to the existing ones. As part of this effort, it is collaborating with one of India’s major anti-venom manufacturers — Bharat Serums and Vaccines (BSV). This may hold the key to finding a more effective ASV, which can contain the high snake bite fatality and related disability rates in India.
The country is home to over 270 species of snakes, 60 of which are venomous. But a four-year, multi-centric study of Evolutionary Venomics Lab under the Centre for Ecological Sciences of IISc found out that the available ASVs are ineffective in treating most region-specific medically important but neglected snake bites. Existing products are only applicable for the Big Four snake species— the spectacled cobra, common krait, Russell’s viper, and saw-scaled viper whose venom collection is limited only to two districts in Tamil Nadu.
Once Dr Kartik Sunagar, who is heading the research, completed the study, IISc tied up with BSV to make test batches of regionally-effective anti-venoms. He said that the new ASVs, for the first time, will offer protection against all clinically important snake species found in the given region, including various species/sub-species of vipers, kraits, and cobras.
Citing an example, he said the ASVs will offer protection against the Sind krait, which the lab’s research has shown to be India’s most toxic snake species. “The range of Sind krait extends all the way from Punjab to northern parts of Karnataka in the west of the country. Considering the toxicity of this snake, and the fact that it is potentially responsible for a large number of deaths in these regions, it is imperative to produce ASVs that are effective against them,” Dr Sunagar added.
The test samples will evaluate the effectiveness of regional ASVs in comparison to conventional products. “For example, this research will inform us whether we need to produce a single anti-venom for western India, from Rajasthan to Karnataka, or if we need to produce specific anti-venoms for specific regions (for example Rajasthan, Maharashtra and Karnataka),” he said. Once proven, IISc will conduct similar studies across other regions of India and identify regions that need a specific antivenom.
According to a BSV statement, “There is an urgent need for the development of region-specific anti-venom in the country. This partnership aims at driving the anti-venom development that will deliver efficient snake bite therapy to save lives, limbs and livelihoods of those in need of the treatment.”
Annually, snakebites cause over 50,000 deaths in India. And four times as many suffer from permanent loss-of-function injuries, such as amputations. The IISc’s study on the spectacled cobra and Russell’s viper, for the first time, highlighted how significant venom variation can be across populations. For example, while some populations of cobras, such as in West Bengal, were extremely neuro-toxic (causing paralysis) the population from Rajasthan was extremely cytotoxic or flesh-dissolving. “This resulted in the anti-venoms completely failing to save mice that were injected with the venom of Rajasthan cobras. In these studies, we found that the anti-venom was particularly ineffective in the northern Indian regions,” Dr Sunagar said.
Asked if they will have to get the nod from the Drugs Controller General of India to roll out the new anti-snake venom in the market, he said that it wouldn’t be applicable to “regional anti-venoms” as the production strategies are identical to the ones that are being used. The only difference between existing and regional anti-venoms would be the source of venoms.
“Hence, human clinical trials are not necessary for regional anti-venoms. While I say that, it is important to clinically test the effectiveness of Indian anti-venoms – regional or conventional – through human clinical trials as they inform us of their real-world performance. This is even more important considering that Indian anti-venoms have never been clinically validated in humans,” he said.(The author is Director of Trade and Investment of Grupo 108 and Independent Latin America and Caribbean Countries Market Expert. Email: intolatam@gmail.com. Views expressed are personal and do not reflect the official position or policy of the Financial Express Online.)
Snake antivenom in India ineffective in treating bites from most species, study finds
The study was conducted by scientists at IISc’s Evolutionary Venomics Lab along with herpetologists Gerard Martin and Romulus Whitaker.
Bengaluru: The only available antivenom in India may be ineffective in treating bites from most of the lethal snake species found in the country, a study has found.
The research, published Friday in the PLOS Neglected Tropical Diseases, was conducted by scientists at the Indian Institute of Science’s Evolutionary Venomics Lab along with herpetologists Gerard Martin and Romulus Whitaker.
India accounts for nearly 46,000 deaths from snakebites each year, the highest in the world, besides around 1,40,000 grave injuries.
The antivenom in use in India, a polyvalent derived from the venom of the four snake species responsible for most bites in the country, was found to be ineffective in neutralising the venom of less known but medically important species such as kraits, the study said.
“Only about 60 species of snakes can potentially inflict lethal bites or severe morbidity but we do not fully comprehend the magnitude of the medical problem imposed by these neglected snake species in India,” Kartik Sunagar, the paper’s lead author, told ThePrint.
Explaining the lack of detailed statistics on snakebites in India, Sunagar said, “It is not a notifiable disease in India, victims often don’t see the snake, venom detection kits are mostly unavailable and clinicians are not trained to identify species.”
Old manufacturing protocols
Despite the problems plaguing the crucial drug, the polyvalent antivenom continues to be derived through a protocol several decades old.
Antivenom is typically known to work when it has been derived from the same species that caused the bite. In India, the process of antivenom manufacture is problematic because it is derived from the venom of the ‘Big Four’ snake species and is only effective against bites from these.
The country, however, is home to over 270 species, 60 of which are venomous.
Snakebites are among the world’s most neglected tropical diseases (NTD), with the World Health Organisation (WHO) estimating that there are more than 27 lakh cases annually and 5 lakh deaths or permanent disabilities. Half of the snakebite-caused deaths occur in India alone.
How is antivenom created
Antivenom is made by injecting venom in small doses into the bodies of animals such as horses, cows and goats. The antibodies produced by these animals against the venom are then collected and processed for purification.
The first antivenom was developed in 1895 by Albert Calmette against the Indian Cobra.
In India, only horses are used for antibody production and the venom utilised is a combination of those collected from the ‘Big Four’ species — the spectacled cobra, common krait, Russell’s viper, and saw-scaled viper. Manufacturers source most of the venom from the Irula Snake Catchers Industrial Cooperative Society in Tamil Nadu.
An antivenom binds with the venom proteins and neutralises their toxicity. However, each species and sub-species of snakes displays variation in the protein composition of venom.
‘Weak antivenom’
Researchers in this study collected venom from 45 individual snakes belonging to seven different species, including the Sochurek’s viper, Sind krait, banded kraits, and monocled cobra.
The study involved an ‘in vitro’ exam to determine the antivenom’s binding ability and then tested its efficacy on mice.
The researchers found that the polyvalent antivenom made by four different manufacturers in India displayed varying degrees of binding abilities. They also found that, in mice, the antivenom failed to neutralise venom from the Sind krait and two populations of the monocled cobras, which are all close relatives of the ‘Big Four’.
They also found that venom from Sind krait was six times more potent than the common krait’s and 40 times more lethal than the common cobra’s.
Variations in the composition of venom across geographical regions was to blame for the drug’s inefficacy, the study said. The antivenom compositions were also found to contain up to 10 times fewer antibodies than some of their global counterparts.
The drug was also unable to counter venom produced by the north Indian common krait — one of the ‘Big Four’ species for which it was created.
“Crude venom is used for immunisation of horses… it contains other non-toxic proteins found in the saliva of the snake, as well as toxins that are not dangerous to humans but only effective against the natural prey or predators of snakes. In addition, the venom may also contain impurities,” said Sunagar.
Venom composition in snakes is also highly dependent on creatures they prey upon. There have been instances when toxicity of venom has evolved in varying degrees to counter non-mammalian prey. Venom of the banded krait and Sochurek’s viper, for example, are least potent against mammals but fatal for non-mammals.
“Therefore, immunising horses with this raw cocktail of venom for antivenom production subsequently reduces the abundance of therapeutically important antibodies,” Sunagar added.
More venom collection centres needed
Scientists are yet to map the variations of toxicity in venom from snakes belonging to various species and geographical locations, and variations within the same species in similar regions.
If the antivenom isn’t effective against common kraits in northern India, does it work effectively against those in other regions? Do snakes kept in captivity by antivenom manufacturers differ in venom composition to those in the wild?
Sunagar and his team at the Evolutionary Venomics Lab hope to answer many such questions.
“Producing a recombinant antivenom that selectively targets all of the clinically important toxins would be a step forward for the snakebite therapy in India,” he said.
For the time being, Sunagar suggests the government invest in venom collection centres across the country to manufacture regionally effective antivenoms. It would increase efficacy of the drug against bites by different varieties of snakes.(https://theprint.in/science/snake-antivenom-in-india-ineffective-in-treating-bites-from-most-species-study-finds/331923/)
Antivenom therapy in India is bitten by shortcomings: Study
Venomous Menace: Snakebite Treatments Are Failing in India
The most widely used antivenom is not effective against the venom of several common snakes.Home to more than 60 species of venomous snakes, India bears the world’s largest burden of death and disability caused by snakebites. The gold-standard treatment is an antivenom raised against the combined venom of four widespread species that are responsible for most attacks, collectively referred to as the “big four.” Although this critical treatment routinely saves lives, a new study published in early December in PLOS Neglected Tropical Diseases shows that it comes up short against the venoms of other Indian snakes whose bites can be deadly.
Kartik Sunagar, an evolutionary biologist at the Indian Institute of Science in Bangalore, co-authored the study. It examined the venom compositions of Sochurek’s saw-scaled viper, two populations of monocled cobras, the banded krait and the Sind krait—perhaps the most toxic snake in India—and three of their closest big-four cousins. Working with herpetologists Romulus Whitaker of the Madras Crocodile Bank Trust and Center for Herpetology in Chennai, and Gerard Martin, founder of a conservation organization called the Gerry Martin Project in the Mysore District, the researchers identified 45 snakes and extracted their venom. Once they identified the proteins and toxins in the samples, they used mouse models to test their toxicity and the efficacy of existing antivenoms. Alarmingly, the team found that the most widely marketed antivenom failed against venoms from both populations of the monocled cobra, the Sind krait and northern populations of the common krait.
Conventional antivenoms are produced by immunizing animals such as horses or sheep with venom and collecting the antibodies they produce, a protocol that has remained unchanged for more than a century. In India, horses are injected with a cocktail of venoms from Russell’s viper, the saw-scaled viper, the spectacled cobra and the common krait, sourced exclusively from snakes in the southern state of Tamil Nadu. This is a problem.
Studies have shown that snake species’ venom compositions can differ dramatically. “We treat snakebites as one medical emergency. But the reality is, if you’re bitten by a snake like a cobra, then you might suffer neurotoxic effects that lead to respiratory paralysis, [and] if you’re bitten by the Russell’s viper, you may have a completely different variety of syndromes such as hemorrhage or bleeding disorders,” says Nick Casewell, who works on animal venoms at the Liverpool School of Tropical Medicine in England and was not part of this study.
The venom of the same species can even vary among different geographical populations. In India, species are distributed widely across the vast subcontinent. Sunagar and his colleagues found that whereas the venom of one monocled cobra population targets the nervous system, the other population’s venom is rich in toxins that cause cell and tissue damage. “The venoms of local, medically relevant snakes must be used to produce antivenoms that will work more effectively in that region,” Sunagar says, adding that more government funding and attention would go a long way. Sunagar and his colleagues are now working with antivenom manufacturers to develop and test such region-specific formulations.
Several global efforts are underway to upgrade existing snakebite treatments and discover novel therapies. Sunagar and Casewell, along with scientists from Kenya and Nigeria, are part of a year-old consortium that aims to develop a new type of antivenom to counteract venoms from Africa and India. The idea is to design treatments that will specifically target the toxins in venom. At the moment, only 15 percent of antibodies in an antivenom are specific to toxins, Casewell says; the rest are directed toward parasites or germs in the involved animal’s environment. Instead of just collecting antibodies from the animals’ blood, he and his colleagues will gather the cells that produce them and grow these in the lab, using them to generate a synthetic “library” of antibodies. “We can then choose just those few antibodies that neutralize lots of different toxins and use that as a treatment moving forward,” he explains. This cocktail of select antibodies will likely also be safer for patients, many of whom suffer adverse reactions to the numerous foreign proteins from other animals that make up the conventional antivenom, he says.
Another promising treatment approach involves a small molecule called varespladib, identified by Matthew Lewin, founder of Bay Area startup Ophirex. Lewin and his colleagues have shown that the compound is very potent against a virulent component of venom called sPLA2, found in abundance in many of the world’s venomous snakes. Varespladib is also small enough to penetrate brain tissue, which conventional antivenoms fail to reach. The team is preparing to perform clinical trials of the compound soon.
Researchers expect that collectively, their research will yield multi-pronged solutions that can be used in isolation or in tandem to combat snakebites worldwide. “However,” Sunagar says, “with 200,000 people affected by snakebites annually, we cannot wait for the next-generation antivenoms. What India needs immediately are antivenoms that are effective against local snake populations in different regions.”(https://www.scientificamerican.com/article/venomous-menace-snakebite-treatments-are-failing-in-india/)
On World Snake Day, call to develop effective antivenom
The antivenom available in India cannot counter the venom of the green pit viper behind most snakebite cases in the northeast
The green pit viper may not be more lethal than Russell’s viper, the saw-scaled viper, the spectacled cobra or the common krait. But what it injects from its poison glands often renders the polyvalent antivenom derived from the venom of the other four ineffective.
The monocled cobra, the banded krait, the lesser black krait, the great black krait, the mountain pit viper and the redneck keelback are among 15 venomous snakes out of 64 species recorded so far across the northeast. Most of the snakebite cases in the region involve different species of the green pit viper, bringing up the other venomous snakes.
The hemotoxic venom a green pit viper injects prevents the blood in the body of a bitten person from clotting, leading to internal bleeding. “Hardly any of the cases we have been fatal. But we cannot draw an inference as very little data on such snakebites are available,” Guwahati-based herpetologist Jayaditya Purkayastha said.
According to Surajit Giri, one of very doctors dealing with snakebite cases in the northeast, the antivenom derived from the ‘Big Four’ snakes are ineffective on the toxin injected by the green pit viper, or for that matter, most other venomous snakes found in the region.
The doctors referred to a 2019 study titled “Beyond the Big Four” to underline the reason. The study said venoms across these species and subspecies are extremely diverse, citing the example of the monocled cobra from West Bengal whose venom contained mostly neurotoxins while the same species from Arunachal Pradesh had cytotoxins in its venom.
Neurotoxin is a poison that acts on the nervous system. Cytotoxins kill the cells in a body.
“The antivenom we have in India is a case of one size fits all. Apart from depending on four species of snakes, experts have pointed out that about 80% of all polyvalent antivenom are derived from snakes caught in one district of Tamil Nadu,” Mr. Purkayastha said.
There have been cases of doctors administering more than 50 vials of antivenom for each cobra or krait bite in northern India because of low efficacy of the standard antidote, putting patients at risk of developing adverse reactions.
Herpetologists and clinicians in the northeast have thus called for developing a range of antivenoms that can do what the four-snake concoction cannot. They chose World Snake Day – celebrated every July 16 to raise awareness about snakes and their importance in our ecosystem – to make the appeal.
Data deficient
The true global burden of snakebit is not known due to the lack of standardised reporting or underreporting, but available data say there are more than 1.4 million cases resulting in 1,25,000 fatalities annually.
More than 46,000 people – the highest on earth – die and thrice the number are disabled due to snakebites every year with 77% of the victims dying outside healthcare facilities, indicating that people go for alternative treatment.
Most snakebite cases in India are reported between June and September and 58% of the victims are farmers and labourers.
About 1,800 vials of antivenom are sold from Guwahati – supplied to various parts of the northeast – every year. This, experts said, is negligible in a region where snakebite is common and points to limited use of antivenom due to its ineffectiveness.( https://www.thehindu.com/news/national/other-states/on-world-snake-day-call-to-develop-effective-antivenom/article65646813.ece)
Snake venom can now be made in a lab and that could save many lives
If you’re unlucky enough to have a poisonous snake sink its fangs into you, your best hope is an antivenom, which has been made in the same way since Victorian times.
It involves milking snake venom by hand and injecting it into horses or other animals in small doses to evoke an immune response. The animal’s blood is drawn and purified to obtain antibodies that act against the venom.
Producing antivenom in this way can get messy, not to mention dangerous. The process is error prone, laborious and the finished serum can result in serious side effects.
Experts have long called for better ways to treat snake bites, which kill some 200 people a day.
Now – finally – scientists are applying stem cell research and genome mapping to this long-ignored field of research. They hope it will bring antivenom production into the 21st Century and ultimately save thousands, if not hundreds of thousands, of lives each year.
Researchers in the Netherlands have created venom-producing glands from the Cape Coral Snake and eight other snake species in the lab, using stem cells. The toxins produced by the miniature 3-D replicas of snake glands are all but identical to the snake’s venom, the team announced Thursday.
In a parallel breakthrough, scientists in India have sequenced the genome of the Indian cobra, one of the country’s “big four” snakes that are responsible for most of the 50,000 snakebite deaths India sees a year.
“They’ve really moved the game on,” said Nick Cammack, head of the snakebite team at UK medical research charity Wellcome. “These are massive developments because it’s bringing 2020 science into a field that’s been neglected.”
From cancer to snake venom
Hans Clevers, the principal investigator at the Hubrecht Institute for Developmental Biology and Stem Cell Research in Utrecht, never expected to be using his lab to make snake venom.
A decade ago, he invented the technique to make human organoids – miniature organs made from the stem cells of individual patients. They’ve allowed doctors to test the specific effects of drugs safely outside the body, something that has revolutionized and personalized areas such as cancer treatment.
So why did he decide to culture a snake venom gland?
Clevers said it was essentially a whim of three PhD students working in his lab who’d grown bored of reproducing mouse and human kidneys, livers and guts. “I think they sat down and asked themselves what is the most iconic animal we can culture? Not human or mouse. They said it’s got to be the snake. The snake venom gland.”
“They assumed that snakes would have stem cells the same way mice and humans have stems cells but nobody had ever investigated this,” said Clevers.
After sourcing some fertilized snake eggs from a dealer, the researchers found they were able to take a tiny chunk of snake tissue, containing stem cells, and nurture it in a dish with the same growth factor they used for human organoids – albeit at a lower temperature – to create the venom glands. And they found that these snake organoids – tiny balls just one millimeter wide – produced the same toxins as the snake venom.
“Open them up and you have a lot of venom. As far as we can tell, it’s identical. We’ve compared it directly to the venom from the same species of snake and we find the exact same components,” said Clevers, who was an author of the paper that published in the journal Cell last week.
The team compared their lab-made venom with the real thing at the genetic level and in terms of function, finding that muscle cells stopped firing when exposed to their synthetic venom.
Cells and DNA, not horses
The current antivenoms available to us, produced in horses not humans, trigger relatively high rates of adverse reactions, which can be mild, like rash and itch, or more serious, like anaphylaxis. It’s also expensive stuff. Wellcome estimate that one vial of antivenom costs $160, and a full course usually requires multiple vials.
Even if the people who need it can afford it – most snakebite victims live in rural Asia and Africa – the world has less than half of the antivenom stock it needs, according to Wellcome. Plus antivenoms have been developed for only around 60% of the world’s venomous snakes.
In this context, the new research could have far-reaching consequences, allowing scientists to create a biobank of snake gland organoids from the 600 or so venomous snake species that could be used to produce limitless amounts of snake venom in a lab, said Clevers.
“The next step is to take all that knowledge and start investigating new antivenoms that take a more molecular approach,” said Clevers.
To create an antivenom, genetic information and organoid technology could be used to make the specific venom components that cause the most harm – and from them produce monoclonal antibodies, which mimic the body’s immune system, to fight the venom, a method already used in immunotherapy treatments for cancer and other diseases.
“It’s a great new way to work with venom in terms of developing new treatments and developing antivenom. Snakes are very difficult to look after,” Cammack said, who was not involved with the research.
Clevers said his lab now plans to make venom gland organoids from the world’s 50 most venomous animals and they will share this biobank with researchers worldwide. At the moment, Clevers said they are able to produce the organoids at a rate of one a week.
But producing antivenom is not an area that pharmaceutical companies have traditionally been keen to invest in, Clevers said
Campaigners often describe snakebites as a hidden health crisis, with snakebites killing more people than prostrate cancer and cholera worldwide, Cammack said.
“There’s no money in the countries that suffer. Don’t underestimate how many people die. Sharks kill about 20 per year. Snakes kill 100,000 or 150,000,” said Clevers.
“I’m a cancer researcher essentially and I am appalled by the difference in investment in cancer research and this research.”
Venom is a complex cocktail
One challenge to making synthetic antivenom is the sheer complexity of how a snake disables its prey. Its venom contains several different components that have different effects.
Researchers in India have sequenced the genome of the Indian Cobra, in an attempt to decode the venom.
Published in the journal Nature Genetics earlier this month, it’s the most complete snake genome assembled and contains the genetic recipe for the snake venom, establishing the link between the snake’s toxins and the genes that encode them. It’s not a straightforward cocktail – the team identified 19 genes out of 139 toxin genes as the ones responsible for causing harm in humans.
“It’s the first time a very medically important snake has been mapped in such detail,” said Somasekar Seshagiri, president of SciGenom Research Foundation, a nonprofit research center in India.
“It creates the blueprint of the snake and helps us get the information from the venom glands.” Next, his team will map the genomes of the saw-scaled viper, the common krait and the Russell’s viper – the rest of India’s “big four.” This could help make antivenom from the glands as it will be easier to identify the right proteins.
In tandem, both breakthroughs will also make it easier to discover whether some of the potent molecules contained in snake venom are themselves worth prospecting as drugs – allowing snakes to make their mark on human health in a different way to how nature intended – by saving lives.
Snake venom has been used to make drugs that treat hypertension (abnormally high blood pressure) and heart conditions such as angina.
“As well as being scary, venom is amazingly useful,” Seshagari said.( https://edition.cnn.com/2020/01/29/health/snake-venom-lab-organoids-stem-cell-scn/index.html)
SNAKE ANTIVENIN
Lyophilised, polyvalent, enzyme refined immunoglobulinis(equine)
Reconstitute with 10 ml of sterile water for injection I.P.
On reconstitution contains, equivalent of purified equine globulins
1 ml of reconstituted serum neutralizes
0.6 mg of Indian Cobra (Naja naja) venom
0.45 mg of Common Krait (Bungarus caeruleus) venom
0.6 mg of Russell’s Viper (Vipera russelli) venom
0.45 mg of Saw scaled Viper (Echis carinatus) venom
Phenol I.P. as a preservative not more than 0.25% w/v
| Polyvalent Snake Antivenin Serum I.P.(ASVS) | |||||||||||||
| A freeze dried polyvalent snake antivenin useful in case of poisonous snake bites. Lyophillsed form it can be stored at comparatively high room temperature. On reconstitution, it is a solution of purified antibodies prepared from equine blood.Available in vials, along with water for injection, to reconstitute 10ml. | |||||||||||||
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| Packing : | |||||||||||||
| 1,5,10,20 vials with equal no. sterile water for injection ampoules. | |||||||||||||
| Indication : | |||||||||||||
| Poisonous snake-bite treatment. | |||||||||||||
| Dosage : | |||||||||||||
| Initially 1-2 vials further depending upon degree of envenomation, preferably under the cover of anti histaminics, sensitivity test advised. | |||||||||||||
| Adverse Drug Reaction : | |||||||||||||
| Occasional sensitivity reactions acute anaphylactic shock which needs immediate treatment. | |||||||||||||
| Precautions : | |||||||||||||
| · Sensitivity tests should be done whenever possible before administration.
· Antihistaminecs and treatment for anaphylactic shock should be kept ready. |
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| PREAMBLE: | |||||||||||||
| Snake Antivenin is of equine origin derived from the plasma of the Horse, Ponies, Mules etc. that have been hyperimmunized against the venoms of the four most common venomous snakes of India. Namely 1) Indian Cobra (Naja naja ) 2) Common krait (Bangarus caeruleus) 3) Russell’s Viper (Vipera russellii ) and 4) Saw Scaled Viper (Echis carinatus). Serum Obtained from the plasma contains Purified,enzyme-refined and concentrated specific heterologous immunoglobulins. It is used as a passive immunising agent and affords Protection to the suseptible victims against the bites of the snake species mentioned above.1 ml. of the reconstituted Snake Antivenin neutralises 0.6 mg. of the Indian Cobra Venom,0.45 mg. of common krait Venom, 0.6 mg. of Russell’s Viper venom and 0.45 mg. of the saw-scaled viper Venom. |
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| Direction for Use : | |||||||||||||
| A. Indications : | |||||||||||||
| The Snake Antivenin is indicated for all bites caused by 1. Indian Cobra 2. Common krait 3. Russell’s viper and 4. Saw-scaled viper, where the patients with clinical signs and symptoms of envenomation. |
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| B. Precautions to be observed before administration of snake Antivenin: | |||||||||||||
| · Elicit history of familial allergic disorders such as asthma, eczema, drug allergy from the patient.
· Whether he had received earlier, injection of serum such as anti-tetanus serum, anti-diptheria serum etc. · Carry out the sensitivity test on the patient. Inject subcutaneously 0.1ml of the serum diluted 1:10. Observe the patient for 30 min. for local or general reactions, if any. In the absense of adverse reaction, administer the requisite dose by the chosen route of injection. · Keep handy injection Adrenaline (Epinephrine) 1ml of 1:1000 along with antihistamines and steroids to meet any emergency arising out of sensitivity reactions. · |
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Snake antivenom is manufactured by the following organizations:
· Haffkine Bio-Pharmaceutical Corporation Ltd., Mumbai
· Central Research Institute, Kasauli
· Bharat Serums and Vaccines Ltd., Mumbai
· Biological E Ltd., Hyderabad
· VINS Bioproducts Ltd., Hyderabad
· Serum Institute of India Ltd., Pune
· Kings Institute of Preventive Medicines, Chennai
Bengal Chem, country’s first maker of snake anti-venom, to resume production:
The country’s first anti snake venom-maker, Bengal Chemicals & Pharmaceuticals, plans to resume production of its flagship offering, the anti-snake venom serum.A proposal in this regard is awaiting clearance from the centre. Set up by the renowned Bengali chemist and nationalist entrepreneur Acharya Prafulla Chandra Ray in Kolkata in 1901, the company – nationalised in 1980 – was forced to stop production of the life-saving serum in 2006-07 following the lack of good manufacturing practice clearances and ageing donor animals, including horses.
The unit was then producing 100,000 vials annually.According to PM Chandraiah, Managing Director and Director (Finance), the Bengal Chemicals & Pharmaceuticals will invest approximately Rs 30 crore as it plans to buy donor animals (horses) and have a new manufacturing set-up. The company’s Kolkata unit will be leveraged for the manufacture of antivenom. The target is to have an annual capacity of 400,000 vials.
Typically, antivenoms are produced using a donor animal – in most cases a horse –which is injected with the snake venom so that it produces a neutralising antibody.
Then, the blood is collected and purified to make the serum.
“We intend to resume production of the anti-snake venom serum over the next three years. Proposals have been sent across to the Ministry,” he told BusinessLine in an interview.
Failed Efforts
Bengal Chemical had started work on upgrading its anti-snake venom manufacturing facilities, in 2008-09. Construction work had begun, but was subsequently put on hold for want of funds.
Bengal Chemical reported its last profit in the early 1950s. Since then, it has remained in the red. However, its finances started improving with restructuring and modernisation of production facilities, improved sales and marketing, and better administration. The company reported its first profit (since nationalisation) of Rs 5 crore in FY17. In FY18, profits doubled to Rs 10 crore.
Chandraiah expects the company to report a profit of Rs 15-20 crore this fiscal, making it the highest profit-making pharma PSU.
“Today, we are profitable and can resume production of the anti-snake venom serum on our own if given the go-ahead” he said. So far, profits have been driven primarily by its home-care, personal care and pharmaceuticals verticals through heritage brands like ‘Phenol’ (floor cleaning phenyl brand), Aqua Ptychotis (indigestion syrup made from ajwain seeds) and Cantharidin hair oil.The pharma division manufactures generic drugs and antibiotics supplied mostly to government hospitals.
Bengal Chemicals’ numbers, Chandraiah said, are expected to get a boost once the serum making starts.
The life-saving drug is in short supply with a handful of private manufacturers dominating the market.
“Anti-snake venom serum sales are expected to contribute at least 15-20 per cent to our bottom-line. The market scope is huge,” he said.
The average cost of a serum varies between ₹700-800 per dosage; and at least 10-15 doses are required depending on the intensity and the type of snake bite.
“If we achieve scale, we may look at exports to neighbouring countries where there is demand,” said Chandriah.
India, as per the World Health Organisation (WHO), accounts for nearly 50 per cent of the global snake bite cases and reported deaths. The most common snake bites include those from Indian cobra, Russel’s viper, scaled viper and common krait.
Various estimates suggest that there are 2.8 million snake bite victims annually, of which 46,900 people die.
In March 2016, Union Minister of Health and Family Welfare JP Nadda told Parliament that there were 3,252 deaths in three years (2013-2015) across India due to snake bites.
Of those, 483 deaths were reported from West Bengal alone.(https://www.thehindubusinessline.com/companies/bengal-chem-countrys-first-maker-of-snake-anti-venom-to-resume-production/article25853776.ece)
Irulas got Permission to Sell Snake Venom
Sales worth R57 lakh to make anti-venom allowed; forest department also gives society licences to catch snakes
CHENNAI: Coming as a huge relief, the State forest department has permitted the Irula Snake Catchers’ Industrial Cooperative Society to sell venom worth Rs 57 lakh, and issued licences for the Irulas to catch snakes. A Government Order (GO) to this effect was issued on Monday.
The livelihood of hundreds of Irulas, who are globally renowned snake catchers, had been crippled as the forest department delayed granting permission to catch snakes. Besides, orders for venom were gathering dust at the Office of the Chief Wildlife Warden.
TNIE recently highlighted the plight of Irulas in a report titled ‘TN govt’s apathy ‘killing’ India’s top snake venom producer?’, following which the government reportedly asked the society to let its members start catching snakes from March 26. Chief Wildlife Warden Syed Muzammil Abbas, who took charge only recently, has issued sale permits for 224 g of snake venom worth Rs 57 lakh. The Irula society is India’s leading supplier of snake venom, with a turnover of about Rs 4 crore annually. Of late, sales and profits have dwindled. This year, the society only managed sales to the tune of Rs 30 lakh.
An official from the society said their struggles are due to the forest department’s reluctance to let them catch their full quota of snakes. The Madras High Court in 1994 allowed capturing of 13,000 snakes annually to extract venom, but in the past few years the forest department hasn’t permitted the Irulas to catch more than 5,000 snakes. In 2021 too, they were only allowed to catch 5,000 snakes. This approach, the official said, was leading to a venom shortage. The society now only has venom of spectacled cobras and Russell’s vipers. Companies manufacturing anti-venom require venom of the common krait and saw-scaled viper too. If Irulas are not allowed to catch more snakes, companies might buy venom from unauthorised third parties to meet the need, the official said.
The Irula Society is the only authorised supplier of venom to produce anti-snake venom in India. Studies show that the composition of snake venom within a species can vary based on location. This has brought about concerns that venom from a specific geographical area may not effectively neutralise venom of the same species from another region. In such cases, larger doses of anti-snake venom serum may be required.
A recent study indicates snakebite envenoming in India has resulted in 1.2 million deaths from 2000 to 2019 i.e. an average of 58,000 deaths per year. About 70 per cent of these snakebites occurred in nine States — Bihar, Jharkhand, Madhya Pradesh, Odisha, Uttar Pradesh, Andhra Pradesh, Telangana, Rajasthan, and Gujarat.(https://www.newindianexpress.com/cities/chennai/2022/mar/29/at-last-irulas-get-permission-to-sell-venom-2435340.html)
Indian Snakebite Project
SNAKEBITE: INDIA’S MOST NEGLECTED TROPICAL ‘DISEASE’
Indian Russell’s viper from Bangalore, Karnataka. This species bites thousands of Indian men, women and children every year, but Indian-made antivenoms may not be effective against all populations of the this snake.
India’s first national survey of the causes of death, the Million Death Study, undertaken in 2001-03 by the Registrar General of India and the Centre for Global Health Research gives an estimate of 46,000 annual deaths by snakebite in the country whereas the Government of India’s Central Bureau of Health Intelligence reports only 1,350 deaths each year for the period 2004 to 2009. This massive statistical disparity has important and urgent implications. There are four species groups of snakes (of the nearly 300 different species in India) primarily responsible for what is likely to be the highest death rate from snakebite in any country in the world, the ‘Big Four’: cobra (four species), krait (eight species), saw-scaled viper (two subspecies) and Russell’s viper. All are widely distributed throughout most of the country although areas like the far Northeast, the Himalayan region and the Andaman’s and Nicobar Islands have distinctive snake fauna.
The venom for producing antivenom comes mainly from the Irula Snake-catchers Industrial Cooperative Society at the Madras Crocodile Bank on Chennai’s East Coast Road. And herein lies one of the problems. Clinicians in other parts of the country are reporting that the antivenom they are using is relatively ineffective in counteracting the effects of a venomous bite. This could be explained by geographic variation in the composition of the venom of a single species.
Since snakebite is a rural problem, primarily affecting India’s farmers, rural labourers and their families it would make sense for antivenom and associated treatment to be available at Primary Health Centers and other rural medical facilities. However, this is often not the case and training in snake identification and snakebite treatment is woefully inadequate.
The Million Death Study puts it in a nutshell: “Snakebite remains an underestimated cause of accidental death in modern India. Community education, appropriate training of medical staff and better distribution of antivenom, especially to the 13 states with the highest prevalence, could reduce snakebite deaths in India.”( https://www.snakebiteinitiative.org/projects-3/snakebite-indias-most-neglected-tropical-disease/)
WORKING TOWARDS SOLUTIONS
The Madras Crocodile Bank Trust and Centre for Herpetology (MCBT), in collaboration with scientists at the Indian Institute of Science (IIS), Vellore Institute of Technology (VIT), Ashoka Trust for Research in Ecology and the Environment (ATREE), the Australian Venom Research Unit (AVRU) from the University of Melbourne and the Global Snakebite Initiative have begun an ambitious project working with government and antivenom manufacturers to revolutionise the production of snake antivenoms for use in India. This multi-component project will combine science, conservation, and improved production of antivenom starting materials with preclinical and clinical trials of new antivenom formulations. In addition the project is exploring other key questions, particularly in relation to Russell’s viper which is responsible for many serious and fatal bites. Venom is being collected from different geographic areas around India, quickly frozen using a new GSI-developed protocol and then being studied to examine how effectively it is neutralised by the current Indian antivenoms. Detailed proteomic studies will follow. Results of these studies are also expected to contribute to improving the quality and potency of Indian antivenoms.
Synthetic anti-venom could save thousands of snake bite victims
Researchers in Bangalore discover breakthrough in antidote to the bite of an Indian cobra.Thousands of lives could be saved thanks to a breakthrough in the development of snake antivenin, making it more readily available around the world.Scientists are poised to transform the way antivenin – commonly referred to as antivenom – is mass produced after discovering an alternative method to milking snakes.In early research by the SciGenom Research Foundation in Bangalore, DNA mapping of an Indian cobra was used to find the genetic code to produce serum that neutralises a poisonous snakebite.
“We believe the Indian cobra reference genome and the analysis presented will facilitate innovations in antivenin development,” researchers said in a report published in the Nature Genetics journal.
The production of antivenin has not changed since the late 1800s, until now.
Traditional methods involve extracting venom from a snake, which is then injected into a horse. Goats and sheep are also sometimes used.
Antibodies then produced in the horse’s blood are harvested so an antivenin serum can be extracted.
New techniques to develop large quantities of antidote are similar to those that produce insulin medication for diabetic patients.
Researchers in Bangalore mapped out a cobra’s 38 chromosomes to identify its venomous toxins, pairing them with the genes they originated from.
The genetic sequence of each poison was placed into bacteria, where it was tracked to see how it interacted with synthetic human antibodies.
The antibodies that stuck to the toxins were selected to form an effective antivenin.
Researchers hope an effective synthetic alternative could be available within three years.
About 100,000 people die from snake bites each year, with almost half a million more losing limbs in rural areas of India and Africa where antivenin is not available.
Although recent research focused on the Indian cobra, similar techniques could be used to develop other serums to treat snake bites from other species.
A vial of antivenin can typically cost more than $2,000 (Dh7,500).
Current techniques can be laborious, with a pint of coral snake venom taking three years to extract from more than 69,000 milkings.
The desert horned viper ranks as the most abundant venomous snake in the Arabian peninsular and the most common spotted by UAE pest controllers.
As Dubai expands into more desert areas, sightings are becoming increasingly common.That has encouraged specialist education from the Sharjah wildlife conservation department for Rentokil staff in Dubai to safely handle snakes.
“We don’t exactly know what kind of venom is kept in stock at Rashid Hospital, but we know the most common venomous snakes we come across here in the UAE are the desert vipers,” said Dinesh Ramachandran, a Rentokil safety manager.A viper bite causes serious swelling, acute pain and excessive bleeding or clotting, depending on the blend of toxins.Victims can also experience nausea, abdominal pain, sweating, exhaustion, kidney failure and heartbeat irregularities.
The first two hours after the bite are crucial to ensure antivenin is quickly administered.“We were given advice on what to do in the event of our staff getting bitten during an extraction,” Mr Ramachandran said. “Because of the extra call-outs we were receiving, the training was necessary – but no one has been bitten up to now.”(https://www.thenationalnews.com/uae/science/synthetic-anti-venom-could-save-thousands-of-snake-bite-victims-1.961755)
No snakes required! Synthetic antivenom could be a lifesaver
Getting bitten by a venomous snake is bad news. But the problem doesn’t just end at the snake bite. Antivenom is expensive, not always available in plentiful supplies, and requires a trained health care professional to administer it. That means that, even in scenarios in which a person could receive lifesaving or life-altering treatment following a bite, this is by no means guaranteed.
A Danish startup called Serpentides, spun out of the University of Copenhagen, is making a new type of antivenom that’s easier to produce, significantly lower in cost, and could be used by anyone, even if they don’t have medical training. According to its creators, it could represent a crucial step toward a universal antivenom since it neutralizes a toxin that’s found in 75% of all venomous snakes.
“Current antivenoms are the only treatment today against envenoming, and these antivenoms are all antibody-based,” Brian Lohse, an associate professor in the Department of Drug Design and Pharmacology at the University of Copenhagen, told Digital Trends. “The antibodies are made by immunizing animals like horses, using venom from living venomous snakes. The vast majority of companies and institutions work on different varieties of antibody-based antivenom including a version that can be made in living cells. However these are all made for injection in the veins and demand hospital settings, and so far cannot be administered without a doctor, and also cannot be injected at the bite site.”
The “serpentides” developed by the researchers are synthesized entirely chemically. That means no snake farm, no horse farm, no snake handlers, and no complicated and expensive purification steps. Serpentides are peptide-based, and can therefore be made in any existing chemistry lab, making them cheaper to produce. The peptide-based antivenoms could be carried in the form of an EpiPen-type device and used immediately when a person is bitten.
According to the World Health Organization, more than 400,000 people per year suffer serious consequences as the result of snake bites, while around 140,000 die. This solution could therefore have an important step forward.
“We are in the process of founding [our company at present,]” Lohse said. “We aim to have around five different peptide-based antivenoms in our portfolio, targeting different snake [toxins]. Currently, we have tested in mice, and this is looking promising as stated in [our] patent.”(https://www.digitaltrends.com/news/synthetic-antivenom/)
Compiled & Edited by-Chhotu pathak, snake saver, jamshedpur
Reference-on request
IMAGE-COURTESY-GOOGLE
