Stingrays and the toxicological implications of their defense mechanisms Biology of Toxins - Spring 2009 Presented by: S. Hall, MO. Ishak, F. Namdar, H. Hayes •World-famous "Crocodile Hunter" Steve Irwin, known for seeking out and handling some of the most dangerous animals in existence, died on September 4, 2006, in a shocking accident with a stingray. Six weeks later, a stingray jumped into a fishing boat in Florida and stabbed 81-year-old James Bertakis in the chest. •News agencies have reported that Irwin's encounter was with an Australian bull ray, estimated to weigh about 220 pounds (100 kg). Irwin was snorkeling in about 6 feet (2 meters) of water, filming a new documentary titled "Ocean's Deadliest" off the coast of Australia. Irwin was swimming with one of the larger species of rays out there -- Australian bull rays can be up to 4 feet (1.2 meters) wide and 8 feet (2.4 meters) long -Julia Layton The recent passing away of Steve Irwin brought about the interest to research the stingray toxin. In popular media the true reason of death was not determined. The stingray must have been some how provoked in order for this sad event to have occurred. Photo of Irwin's + a croc. The goal of this presentation is to further explore the toxin that was administered from the stingray and all of its logistics… (i.e. ADAME) Photo of Irwin's boat. If someone as knowledgeable about animals as Steve Irwin died from a stingray, then realistically anybody can. Knowledge of stingrays is especially important due to the fact that they are found in most bodies of water and can potentially cause harm to us. Stingrays are often found in tourist locations They are found both in marine and freshwater habitats throughout the world. Many species are “euryhaline” capable of living in waters of a wide range of salinity. Most of the river stingrays of the family Potamotrygonidae live permanently in freshwater and have lost the ability to survive in marine habitats. Generally speaking they can be found in a variety of different places and regions around the globe. Stingray distribution along Southern US and Mexican coasts. Stingrays are unique elasmobranchs (These fish have cartilaginous skeletons, hard teeth and well developed jaws) which are cousins of the well known shark. There are roughly two hundred species of stingrays in the scientific order Myliobatiformes. Pelagic Two types of stingrays 1) Benthic “bottom” 2) Pelagic “swimming” Benthic Hierarchy of Stingray Families and Subtypes 1) Benthic Example: Atlantic stingray ◦ Often buried in the sand ◦ Rounded, diamond shaped body ◦ Stingray located at the middle, lower third of tail ◦ Bottom feeders (worms, clams, shrimp, crabs, snails) 2) Pelagic Example: Spotted Eagle ray and Manta ray ◦ Active swimmers ◦ Bat-like shape ◦ Stingray located near body, just behind pelvic fans ◦ Most are bottom feeders Defensive mechanism: The “striking” action is purely defensive ( an involuntary response) and not a conscious decision (or attack). Benthic: Stingray located further away from body. Making it a more effective “striking” and defensive weapon. Pelagic: Stingray near base of tail to discourage predators from biting near vital organs. Stingray attacks are defensive. They usually occur when an unknowing human steps on the creature. When the “wings” of the stingray are touched, it thrusts its tail forward, driving its venomous spine into the victim. Benthic stingrays burry themselves and often go undetected. • • • • • NO….not necessarily. Dasyatidae There are two separate families of poisonous stingrays Dasyatidae and Urolophidae. Within these two families there are 4 genera and roughly 35 species which are poisonous or that pose a threat to humans (out of about 200 documented species). Evolutionary biologists have not however, studied the evolution of venom glands and envenomation as a distinct subject. Therefore, it is only possible to speculate how stingray venom glands and delivery systems are related to venom producing functions in more advanced life forms. Urolophidae Fig.1 – shows the basic anatomical parts of a stingray Fig 2 – close up of barb that is found on tail of sting ray The sting apparatus is a modified dermal denticle , with two ventral grooves filled with venom-producing tissue. The venom apparatus is surrounded by a cell-rich covering, or sheath that also may produce lesser amounts of venom. The venom itself is largely a proteinbased toxin that causes great pain in mammals and may also alter heart rate and respiration. Rays vary in size from 10 to 12 inches to 6 feet in width and most measure at least 3 feet long. Their tails are often almost twice as long as their bodies. • • • Stingrays contain a cartilaginous spine surrounded by a thin layer of skin. This skin is called the integumentary sheath and covers two ventrolateral grooves that contain venom glands. When the spine penetrates the victim the glandular sheath ruptures to release the venom into the tissue of interest. Dose (mL/kg): ◦ Generally speaking the administered dosage is dependent upon the size of the stingray in question. ◦ Intra-Venous LD50 of the lyophilized (dried by freezing in a high vacuum) venom is estimated at 28mg/kg. Wet sting: ◦ For the most part when a victim is stung by a ray the barb does in fact contain the ray’s toxin (which would qualify as a wet sting). Dry sting: ◦ Since the serotonin in stingray venom produces severe and immediate onset of local pain, any sting that is relatively free of pain indicates that no actual envenomation occurred and the victim endured a “dry” sting. A sharp inner barb with a thin sheath surrounding it that contains the venom. When the spine is ejected, the barb pierces the venom sac, along with the victim’s skin, and the poisonous slime is introduced into the wound. The venom contains enzymes that cause tissue death. Breathing difficulty, low blood pressure, generalized cramps, inflammation, fainting, headache, weakness, paralysis, and bleeding are a short list of the side effects. Venom causes tissue necrosis, especially to muscle and fat. Even after the apparatus is removed, pieces of the spine and membrane, along with toxin and mucus, can remain, producing a festering chronic ulcer and secondary infection if the wound is not inspected and de-brided meticulously. Orpotrin Phosphodiesterases ◦ Vasoconstrictor peptide (causes arteriolar constriction) ◦ Causes Hydrolysis of phosphodiester bonds in DNA and RNA ◦ Necrosis and tissue breakdown Hyaluronidase ◦ Causes Hydrolysis of hyaluronic acid of extra cellular matrix (diffusion factor) ◦ Has antigenic properties Serotonin 5’-nucleotidase ◦ Causes severe vasoconstriction. (painful severe contractions of smooth muscle) ◦ Platelet activation ultimately leading to plug formation ◦ This is why the victim feels severe pain from the sting ◦ Causes complete degradation of linear nucleic acids ◦ Exonuclease- it breaks the last nucleotide in the end of a polynucleotide ◦ Necrosis and tissue breakdown Hyaluronidase Serotonin Phosphodiesterase HGGYKPTDK (1002 Da) Orpotrin 5’-nucleotidase The sheath may have been previously ruptured, leaving no venom to administer in the most recent sting. The sheath failed to penetrate the wound. The sheath failed to rupture and therefore failing to release the toxins. The stingray had a broken spine. Sea turtle and broken barb from stingray MJA 2001;175:33-34 Cardiac injuries from stingray barbs are rare, even world wide; all but one have been fatal (diagram is from the case that survived cardiac injury from a stingray barb) Stingray barb entered the pericardial sheath causing cardiac tamponade. Patient recovered fully after pericardialcentesis. The effects of the venom may be local and/or systemic. Local effects include severe pain at the site of injury, and tissue necrosis. 5 Systemic effects include nausea, vomiting, salivation, sweating, respiratory depression, muscle fasciculations, convulsions, cramping, abdominal pain, arrhythmias (first-, second-, third- degree heart block, and asystole), myocardial ischaemia, and, rarely, death. Many of the systemic effects have been documented only in patients with viscous penetration, and not in those with peripheral stingray injuries. Local venom effects are usually more troublesome in peripheral stab injuries, but if the barb pierces a vital organ or structure mechanical damage may be more dangerous than the venom effects. Ouch! • Most importantly keep the victim calm! • Remove sting fragments with gloves to reduce the risk of injuring your self. • Wash wound with salt water. • • • Induce injured extremity to warm (43-45 C) water for 40 to 90 minutes (venom is thermolabile or is subject to destruction/decomposition or change in response to heat). Use analgesics, NSAIDS, local anesthetics, muscle relaxants, antihistamines, and corticosteroids for pain, itching, and swelling. Antibiotics are commonly used to fight infection. Serotonin is metabolized to 5-hydroxyindole acetaldehyde by monoamine oxidase and excreted through the kidneys into urine. Phosphodiesterase, 5’nucleotidase, Orpotrin, and Hyaluronidase are also all excreted through the kidneys into the urine. This could pose a problem later on in life in regards to nephrotoxicity as well as damage to the bladder possibly leading to cancer. Humans are rarely killed unless the barb punctures vital organs, such as the heart or lungs. Stingrays main predators are sharks, and the barbs are usually ineffective at killing. Steve Irwin did in fact get stung by a poisonous stingray, however he most likely died before the poison took effect on his body. The serotonin in the toxin does have many effects on cardiac tissue but with the time of death being immediately after the barb was removed it can be determined that platelet activation and vasoconstriction leading to death had yet to have occurred. The stingray punctured Irwin’s chest and lodged itself into his heart. The spine was removed, ultimately causing his death due to hemorrhaging. ◦ If (Irwin) was conscious he would have been in agony," Dr Fry told the Reuters news agency. ◦ Dr Fry states, stingray venom was a defensive weapon similar to that in stonefish, whose poison is generally fatal, but in the stingray's case the toxin was not lethal. It was the serrated barbs on the stingray's tail that would have delivered the fatal injury, he said. ◦ "It's not the going in, it's the coming out," Fry said. "They have these deep serrations which tear and render the flesh as it comes out." ◦ Dr Geoff Isbister, a clinical toxicologist, said that little was known about stingray venom, but he agreed that it was the physical trauma associated with the wound that would have killed Irwin. ◦ "What happened to Steve Irwin is like being stabbed in the heart," Dr Isbister said. ◦ http://www.timesonline.co.uk/tol/news/world/article627862.ece RIP Steve Irwin •Liggins, J.B, An Unusual Bathing Fatality, NZMJ, Vol XXXVIII, 1939 •Grenard, Steve, Stingray Injuries, Envenomation, and Medical Management, http://www.potamotrygon.de/fremdes/stingray%20article.htm •Rodriguez, Hector., Sanchez, Edgar., Mendez, Jose., Stingray Poisoning, A careless Aspect in Mexico, Advances in Environmental Biology, 2(2): 54-62, 2008 •Forrester, Mathias,. Pattern of stingray injuries reported to Texas poison centers from 1998 to 2004, Human & Experimental Toxicology, Issue 24, 639-642, 2005 •Diaz, James H., The Epidemiology, Evaluation, and Management of Stingray Injuries, Journal of the Louisiana State Medical Society, Vol 159 July/August 2007 •Russell FE, Panos TC, Kang LW, et al. Studies on the mechanism of death from stingray venom: a report of two fatal cases. Am J Med Sci 1958;235:566 - 83. •Russell, Findlay E., Stingray Injuries, Public Health Reports (1896-1970), Vol 74, No 10 (Oct., 1959), pp 855-859 •Gussow, Leon MD, Stingray's Poisonous Spine is Sharp, Rigid, and Nasty, Emergency Medicine News, Volume 28(11), November 2006, p 12–13 •Evans, R.J, Davies, R.S., Stingray Injury, Accid Emerg Med 1996;13:224-225 •Clark, Richard F., Stingray Envenomation: A Retrospective Review Of Clinical Presentation and treatment in 119 Cases, The Journal of Emergency Medicine, Vol. 33, No. 1, pp. 33-37, 2007 •http://animals.howstuffworks.com/fish/stingray.htm