Poisonings at The Animal Ark

The Animal Ark is a non-profit wildlife sanctuary and animal education center based in Reno, Nevada. They currently house and care for:

Bears including two North American Black Bears

Canids including an Arctic Fox, two Coyotes, a Kit Fox, two Red Foxes, and two Gray Wolves

Felines including an Asian Tiger, two Bobcats, three Canadian Lynx, two Cheetahs, a Jaguar, and two Mountain Lions.

Raptors including an American Kestrel, a Barn Owl, a Great Horned Owl, and a Red Tailed Hawk

Reptiles including two Desert Tortoises

And other animals including an American Badger and a Raccoon.

It was first reported in the media in February 2015, Shere Khan, a 21-year old Bengal Tiger, unexpectedly and rapidly became ill, began exhibiting symptoms of a stroke, and then eventually was euthanized due to his condition.

Shere Khan, the Bengal Tiger

Digital image courtesy of the Animal Ark official website - http://animalark.org/animals/shere-khan/

The next day Animal Ark employees found that two other animals, Maddie the Gray Wolf and Jamar the Cheetah, had become ill with similar symptoms. Maddie was found dead and Jamar was found unresponsive in a coma.

Jamar, the Cheetah

Digital image courtesy of the Animal Ark official website - http://animalark.org/animals/moyo-jamar/

Maddie and Monty, the Gray Wolves

Digital image courtesy of the Animal Ark official website -http://animalark.org/animals/maddie-and-monty/

What happened?

Let’s enter the wonderful world of animal and wildlife forensic toxicology.

Biological specimens were acquired from Maddie and Jamar and toxicological analyses were undertaken. No specimens were acquired from Shere Khan as he had already been buried. While I work in a postmortem toxicology laboratory, the instrumentation used in animal and wildlife forensic toxicology is the same – high performance liquid chromatography (HPLC), gas chromatography with mass spectrometry (GC/MS), headspace gas chromatography with flame ionization detection (GC-FID), liquid chromatography with tandem (triple quadrupole) mass spectrometry (LC/MS/MS), liquid chromatography with time of flight mass spectrometry (LC/ToF or LC/qTOF), etc.

According to media reports, Pentobarbital and Phenytoin were detected in the gastric contents of Maddie. The same substances were found in the urine of Jamar. It was hypothesized that the substances had found their way into the food supply. Through investigation and a little bit of analytical chemistry it was determined that a specific brand of meat given to the animals was contaminated with Pentobarbital and Phenytoin. This news then led to a product recall. As the identified meat company is simply a distributor, this recall led to food product being returned from 10 different zoos and wildlife sanctuaries across the USA.

On a good note, after intensive medical treatment, Jamar the Cheetah survived the ordeal.

So, we exactly are Pentobarbital and Phenytoin?

Pentobarbital is a barbiturate derivative that was first synthesized in 1930 and is considered a Schedule II controlled substance by the United States Drug Enforcement Administration (DEA). It has been used clinically in humans as a short-acting and potent sedative-hypnotic agent for insomnia, pre-anesthesia, and as an anticonvulsant. It has other uses in treating Reye’s syndrome and traumatic brain injury as well in veterinary anesthesia. Pentobarbital is also one of the drugs that has been used in human and animal euthanasia. In November 2013, a Missouri death row inmate died from a dose of the drug during the lethal injection/capital punishment process. Its pharmacological mechanism of action is via GABA receptor potentiation, AMPA and kainate receptor inhibition. The drug also increases the duration or time that chloride ion channels are open. The molecular formula is C11H18N2O3 and molecular weight is 226.2 g/mol. Different forms of the drug exist on the market such as capsules, tablets, suppositories, and solutions for intramuscular and intravenous injection. The most common trade name is Nembutal. It is biotransformed and excreted into the urine as unchanged drug and various hydroxylated and carboxylated metabolites. Symptoms of overdose include impaired psychomotor function, incoordination, slowed or slurred speech, hypotension, severe respiratory depression, and coma. Over the last several decades, the substance has been implicated in many deaths – many of which have been accidental overdoses.

Phenytoin is a hydantoin derivative substance that is used as anticonvulsant drug and was first synthesized in 1908. It is one of the main substances used clinically in treating epilepsy. Phenytoin is also classified as a class 1b antiarrhythmic substance and is sometimes used after cardiac glycoside (i.e. Digitalis, Nerium oleander) poisoning. Its pharmacological mechanism of action is via binding to sodium channels and blocking  the repetitive firing of action potentials. It is not considered to be a Federally controlled substance in the United States and it is available by prescription only. One common trade name is Dilantin. Molecular formula is C15H12N2O2 and molecular weight is 252.2 g/mol. The drug is available in normal release and extended release capsules, liquid suspension, and a solution for intramuscular or intravenous injection. Phenytoin is excreted in the urine primarily as unchanged drug and hydroxylated metabolites. Pentobarbital is formed as a metabolite of the barbiturate derivative, Thiopental, also known as Sodium Pentothal, by desulphuration. Symptoms of overdose include dizziness, drowsiness, slowed and slurred speech, ataxia, hyperglycemia, and blurred vision. Central nervous system and respiratory depression is generally not caused by Phenytoin in normal dosages. When compared to other drugs, Phenytoin has accounted for relatively few deaths, though acute intoxication with the substance is not unheard of.

Now, the next logical question is how did the substances come to be in the food supply? It turns out that the food product was procured from a horse that was euthanized with Pentobarbital and Phenytoin. One common product used in euthanasia of cattle, horses, pigs, dogs, and cats is called Euthasol from LeVet Pharma. It is a solution of 400 mg/mL Pentobarbital and is available in 100 mL or 250 mL volumes. Another version of Euthasol from Virbac USA that is for use in dogs contains 390 mg/mL Pentobarbital sodium and 20 mg/mL Phenytoin sodium.

Truly a sad story. One in which there seems to be no malice or intent, but an oversight into the quality of the product that made it into the food supply.

If you haven't already done so, please visit the Animal Ark's website. They care for some truly beautiful creatures and do some amazing work.

References

Baselt, R.C. (2014) Pentobarbital Disposition of Toxic Drugs and Chemicals in Man. Tenth Edition. Pages 1575-1577. Biomedical Publications, Seal Beach, CA 90740.

Baselt, R.C. (2014) Phenytoin. Disposition of Toxic Drugs and Chemicals in Man. Tenth Edition. Pages 1625-1628. Biomedical Publications, Seal Beach, CA 90740.

 

Pentobarbital. Compound Summary. CID 4737. Pubchem. NCBI. http://pubchem.ncbi.nlm.nih.gov/compound/4737?from=summary

Accessed: 3/23/2014

Dilantin (Phenytoin Sodium injection), FDA approved label, 10/2011. http://www.accessdata.fda.gov/drugsatfda_docs/label/2011/010151s036lbl.pdf

Accessed: 3/23/2014

Digital images of Shere Khan, Jamar, and Maddie and Monty can be found at the Animal Ark official website - http://animalark.org/the-animals/

 

Herbal Incense

If you've never seen 'herbal incense', here it is...

3 grams of 'incense' containing synthetic cannabinoids in a glass bottle.

Digital image taken by ForensicToxGuy (2015). Feel free to use the image if you'd like.

It's not very good.



The Case of Jagger the Irish Setter

At the beginning of March, it was reported that a European show dog, a 3-year old Irish Setter named Jagger, had died with possible suspicious circumstances while competing in the Crufts show in the United Kingdom. Toxicological analyses were completed on specimens drawn during autopsy of the animal. According to the latest reports, the dog’s blood was positive for aldicarb and carbofuran - two substances that should not be found. At all.

What are aldicarb and carbofuran?

 

They are pesticides.

 

Highly potent pesticides.

 

Highly potent and restricted pesticides.
 

 

Aldicarb is a carbamate pesticide primarily used historically as a nematicide in potato production, but also used for the control of pests such as aphids and spider mites. Its IUPAC name is 2-methyl-2-(methylthio)propanal O-(N-methylcarbamoyl)oxime. Molecular formula is C₇H₁₄N2O₂S and molecular weight is 190.2 g/mol. Aldicarb is excreted in the urine primarily as metabolites over approximately 1-10 days. Common urinary metabolites include aldicarb sulfoxide, aldicarb sulfone, aldicarb oxime, and aldicarb nitrile. The trade name for aldicarb is Temik. Its use was banned in 2003 in the European Union.

Carbofuran is a carbamate pesticide that is used on various field crops such as potatoes and corn. It is also effective against soybean aphids. Its IUPAC name is 2,2-dimethyl-2,3-dihydro-1-benzofuran-7-yl-methylcarbamate. Molecular formula is C₁₂H₁₅NO₃ and molecular weight is 221.2 g/mol. Carbofuran is excreted in the urine primarily as 3-hydroxycarbofuran, carbofuranphenol, and 3-ketocarofuranphenol. Trade names for carbofuran include Furadan and Curater. It has been banned for use on all crops grown for human consumption in the European Union since 2008 and in the United States since 2009. Carbofuran is highly toxic to vertebrate organisms. It has an LD₅₀ of 19 mg/kg in dogs and 8-14 mg/kg in rats.

How do these substances act in the body?

From a pharmacological perspective, these substances act via the parasympathetic nervous system (PSNS) as reversible inhibitors of the enzyme acetylcholinesterase (AChE). Typically, acetylcholine (ACh) is released into the neural synaptic cleft and binds to ACh receptors post-synapse, which then relays a signal from the nerve. AChE halts the signal transmission via hydrolysis of ACh into acetate and choline. AChE functions rapidly and has a very high catalytic activity – it has been demonstrated that AChE can degrade about 25,000 molecules of ACh per second. Choline is then taken up by the presynaptic nerve and ACh is constructed from choline and Acetyl Coenzyme A (acetyl-CoA) via another enzyme, choline acetyltransferase.  As AChE inhbitors, aldicarb and carbofuran inhibit or slow down the normal function of this enzymatic pathway of ACh which results in an accumulation of ACh in the neural synaptic cleft. This accumulation can result in devastating consequences for the organism.

The mnemonic SLUDGEM has been used to describe the symptoms and effects of substances such as these:

Salivation stimulation, usually excessively

Lacrimal stimulation, leading to tearing or watery eyes

Urination via urethral internal sphincter muscle relaxation and contraction of the detrusor muscle

Defecation via internal anal sphincter relaxation

Gastrointestinal upset via changes in smooth muscle tone possibly leading to diarrhea

Emesis or vomiting

Miosis or pinpoint pupils

Other adverse effects include blurred vision, bronchial secretions, bronchoconstriction, constriction of the pharynx, bradycardia, diaphoresis, hypotension, hypothermia, vasodilation, tremors, and convulsions.

Many poisonings of humans and animals have occurred using these two substances – some of these have been accidental through occupational exposure or eating contaminated crops, while others have been intentional through attempted suicide or homicide. Many deaths have occurred.

Other substances that act similarly in the body via acetylcholine include nerve agents such as VX and Sarin, organophosphate pesticides such as diazinon and parathinon, the Alzheimer’s disease medications donepezil (Aricept) and galantamine (Nivalin).

In this case - who administered the substances? By all reports, it is unknown at this time, but I'm sure one day, we'll find out who poisoned Jagger the Irish Setter.

 

References

Baselt, R.C. (2014) Aldicarb. Disposition of Toxic Drugs and Chemicals in Man. Tenth Edition. Pages 58-59. Biomedical Publications, Seal Beach, CA 90740.

Baselt, R.C. (2014) Carbofuran. Disposition of Toxic Drugs and Chemicals in Man. Tenth Edition. Pages 351-352. Biomedical Publications, Seal Beach, CA 90740.

Bayer CropScience. Temik material safety and data sheets (MSDS)
https://www.bayercropscience.us/products/insecticides/temik/label-msds
Retrieved 2015-03-17

NIOSH Pocket Guide to Chemical Hazards. Carbofuran. National Institute for Occupational Safety and Health (NIOSH). http://www.cdc.gov/niosh/npg/npgd0101.html

Retrieved 2015-03-17

The Culture High

I recently viewed The Culture High film and it is available to stream on NetFlix. The movie is directed by Brett Harvey, who previously brought us 2007's The Union: The Business Behind Getting High.

Whatever side you stand on legalization or even decriminalization of marijuana in the United States, the film is worth watching at least once.

One thing did feel out of place - around 35 minutes into the film there was a lone commercial/media report on "synthetic marijuana" depicted with a reporter holding up a package of the infamous Mad Hatter branded herbal incense. I don't understand why it was included. Did I miss something? Maybe an error?

Case Report - Pentedrone and Alpha-PVP Associated Death

A Fatal Case of Pentedrone and α-Pyrrolidinovalerophenone Poisoning
Sykutera M, Cychowska M, and Bloch-Boguslawska E
Journal of Analytical Toxicology (2015) doi: 10.1093/jat/bkv011
http://jat.oxfordjournals.org/content/early/2015/03/03/jat.bkv011.full

This case was reported out of the Department of Forensic Medicine, Collegium Medicum of Nicolaus Copernicus University in Poland.

The case involved a 28 year old man who was a known user of “designer drugs” who was transported to the hospital while in asystole. A white powdery substance in a bag labeled “α-PVP” and material safety data sheets (MSDS) for alpha-PVP and pentedrone were retrieved from his jacket pocket.

Resuscitative attempts failed and he was pronounced deceased 30 minutes after arrival. Upon autopsy, pulmonary edema and atherosclerotic lesions of the arteries was documented. Microscopic evaluation of tissues revealed “chronic changes in the heart”, which was defined as “uneven staining pattern of the myocardium, fragmentation and waviness of fibers, perivascular connective tissue growth, intramuscular fibrosis and scarring, disintegration of cardiomyocytes, nuclear disintegration, loss of cross-striations , and thickening of blood vessel walls”. The presence of “hemosiderin-laden pulmonary macrophages” were also observed.

Femoral blood was positive for alpha-PVP (901 ng/mL) and pentedrone (8,794 ng/mL). Liver was positive for alpha-PVP (2,610 ng/g) and pentedrone (100,044 ng/g). Kidney was positive for alpha-PVP (462 ng/mL) and pentedrone (22,102 ng/mL). Brain was positive for alpha-PVP (120 ng/g) and pentedrone (13,48 ng/g). Gastric contents were positive for alpha-PVP (4,190 ng/g) and pentedrone (500,534 ng/g). Cause of death was multiple drug toxicity associated with alpha-PVP and pentedrone use.
_______________

We have talked about alpha-PVP before on this blog in regards to a generic named street drug, gravel.

Gravel: human sacrifice, dogs and cats living together...mass hysteria?

Both pentedrone and alpha-PVP are currently Schedule I controlled substances in the USA.

Foxy's cousin, 5-MeO-MiPT

What did the Fox say? Well, we still aren't really sure.

I've viewed the arrest warrant application and affidavits (provided via the nice folks at Bluelight) that were filed in the Wesleyan University campus, Connecticut “Molly” overdoses.

According to these documents, some specimens confiscated by the authorities field tested positive for amphetamines, but these specimens were analyzed via the CT state crime laboratory and were reported as negative with the exception of a capsule that contained a white-colored powdery rock-like substance. This substance was previously identified as “Molly”. The specimen was identified as 5-Methoxy-N-methyl-N-isopropyltryptamine, otherwise known as 5-MeO-MiPT.

5-MeO-MiPT is a tryptamine substance that is used for its psychedelic and hallucinogenic properties. The substance is a 5-HT1A and 5-HT2A (serotonin) receptor agonist and acts as a serotonin-norepinephrine reuptake inhibitor (SNRI). It may also have monoamine oxidase inhibition (MAOI) activity. It is not considered a controlled substance at the Federal level in the United States, but may be considered a “controlled substance analog” as it is a very close derivative of the scheduled substance, 5-MeO-DiPT, which is known as "Foxy Methoxy" or "Foxy".

According to Alexander and Ann Shulgin’s book TiHKAL: The Continuation (which stands for Tryptamines I Have Known and Loved), the substance is active in 4-6 mg dosages. Commentary in TiHKAL after dosing with 6, 7, 10, and 12 mg oral dosages of the drug are interesting.

If you don't have access to TiHKAL, here are some direct experience quotes:

After a 6 mg oral dosage:

“One hour in and I felt turned on as if a wave passed over my body, and then the wave went back to the ocean, or wherever waves go. I was getting hungry but I didn’t want to go to the kitchen, as I didn’t want to interact with anyone I might encounter. What remained with me the longest was the awareness of vibrations, and what felt best was the stillness.

After a 7 mg oral dosage:

“In one hour I was in a marvelous, sexy place. Everything was shaded with eroticism. Sex was explosive, and in another three hours I was completely ready for the outside, public world.”

After a 10 mg oral dosage:

“Colors on the edges of the wiggles of the eye, a sort of Jessie Allen running design with color contrasts and sparkle. People’s faces were interesting, quite serious, and not completely friendly…this is a definite sense-distorter. I am not completely sure I like it.”

After a 12 mg oral dosage:

“…Warm was comfort and comfort was good. Warm led into a wonderful sexual turn on, where my entire body was alive and alert. This was one hour into the trip. This sexual turn-on was the feeling of a bud about to unfold into a full-blown, beautiful flower, which happened during love making.”

After a 12 mg oral dosage:

“…I have never experienced such an increase of the peristalsis process in moving the burrito through my colon and with each defecation I would become a little more turned on…”

After a 12 mg oral dosage:

“There was a very strange, almost paranoid session of listening to music, about an hour and a half into it…It turned out that I was wrong; the music was strange but of good quality. It was my interpretation that was screwed up.”

After a 12 mg oral dosage:

“My experience with this material was different in its action than anything I had tried in the past – it came on quicker but with much less intensity. I really enjoyed the mellowness, and it sort of waved in and out…”

After a 12 mg oral dosage:

“…Quickly aware and in the second half hour I rapidly shot up to a +++ in a very LSD-like manner, without the visuals. Time was quite slowed down during this come-on. Erotic world was fantastic, explosive, almost scary."

Overall, 5-MeO-DiPT is described by the Shulgins as being easily synthesized, having short lived effects (duration is 4-8 hours) and pretty high potency (4-12 mg oral dosage). Its phsyical side effects are "minimal", but it is a "mixed bag" in regards to its usefuless in experiences.

On a side note, Shulgin also includes a pretty funny story about the initial synthesis of 5-MeO-MiPT and its subsequent publication in literature with a missing co-author. Read it if you have the chance.

My thoughts on all of this?

No MDMA or methylone or other substance is reported as being found or confirmed by the CT state crime laboratory, which is interesting in of itself.

I wonder how much 5-MeO-MiPT was actually in this substance referenced in the documents? Due to the substance's higher potency, may this be a case of way, way too much drug being consumed at one time? MDMA is typically dosed in 50-100 or so mg tablet/capsules. And we see time and time again of large dosages being consumed unknowingly.

IF (and that’s a big IF as this is all speculation on my part) the drug that caused these overdoses was actually 5-MeO-MiPT, then what was the dosage consumed? 5 mg? 15 mg? 50 mg? 100 mg? The only effects described in the documents were "extremely ill", "very ill", "feel very weird", "out of it", "heavy', and "dissociated feeling". Vomiting, feeling hazy, and hallucinations were also mentioned.

It is also interesting, but not surprising (afterall this is a college campus), to note that many other prescription and nonprescription substances were found in the searched areas. These included alprazolam, bupropion, buspirone, cabergoline, carvedilol, coluracetam, dimethyltryptamine (DMT), donepezil, escitalopram, etizolam, hydroxyzine, linisopril, lioresal, methylphenidate, moclobemide, O-acetylpsilocin, olanzapine, oxiracetam, phenylpiracetam, propranolol, rasagiline, sulbutiamine, tamsufine, telmisartan, topiramate, trazodone, and valsartan. A rather eclectic and interesting mix of substances.

Doing a quick literature survey, it seems that there is a single case report from 2007  out of Japan that describes an intoxication after use of a product containing 60% methylone and 38% 5-MeO-MiPT. Not much else is out there in regards to toxicological profile of this substance.

As always, I'll wait to add to more thoughts at a later time when additional information is disclosed (if it ever is).