Genetic identification and subsequent LC-QTOF MS analysis of plant remains (Oenanthe spp.) could prove the cause of an undetermined sudden death.

In recent years, recorded cases related to forensic botany and, in particular, of plant poisoning have become rare. We report on the medicolegal characteristics of an undetermined sudden death (USD) of a woman in which scene there were remnants of a vegetal peeling. After the autopsy, macroscopic findings reported multiorgan failure and requested the investigation of the cause of death. Postmortem blood was firstly investigated on cyanide toxicity presumptively coming from a yucca-like root; however, found cyanide levels were under normality. Because of the lack of morphological features of the encountered plant remains, a genetic nrDNA ITS2 sequence investigation was followed. The resulting DNA sequence could identify the evidence as the water dropwort (Oenanthe spp.) which contains oenanthotoxin, a potent toxin that may be fatal, similar to the more commonly found in hemlock Conium or cowbane Cicuta species. A liquid chromatography-tandem high resolution mass spectrometry (LC-QTOF MS) was later applied to analyse the vegetal extract and stomach content and successfully confirmed the toxin existence. Medicolegal and analytical findings at the forensic laboratory were described, where both biological and chemical techniques could successfully conjugate, as an interdisciplinary research, and explain premortem symptoms and postmortem findings. Present data can be helpful in future investigation on poisoning cases by conjugated polyacetylenes . The present work tries to emphasize the often undervalued plant evidence in legal medicine diagnosis in the context of an unexplained death.

The inside tract: The appendicular, cecal, and colonic microbiome of captive aye-ayes.

OBJECTIVES: The aye-aye (Daubentonia madagascariensis) is famous for its feeding strategies that target structurally defended, but high-quality resources. Nonetheless, the influence of this digestible diet on gut microbial contributions to aye-aye metabolism and nutrition remains unexplored. When four captive aye-ayes were unexpectedly lost to persin toxicity, we opportunistically collected samples along the animals' gastrointestinal tracts. Here we describe the diversity and composition of appendicular, cecal, and colonic consortia relative to the aye-aye's unusual feeding ecology. MATERIALS AND METHODS: During necropsies, we collected digestive content from the appendix, cecum, and distal colon. We determined microbiome structure at these sites via amplicon sequencing of the 16S rRNA gene and an established bioinformatics pipeline. RESULTS: The aye-ayes' microbiomes exhibited low richness and diversity compared to the consortia of other lemurs housed at the same facility, and were dominated by a single genus, Prevotella. Appendicular microbiomes were differentiated from more homogenized cecal and colonic consortia by lower richness and diversity, greater evenness, and a distinct taxonomic composition. DISCUSSION: The simplicity of the aye-aye's gut microbiome could be attributed to captivity-induced dysbiosis, or it may reflect this species' extreme foraging investment in a digestible diet that requires little microbial metabolism. Site-specific appendicular consortia, but more similar cecal and colonic consortia, support the theory that the appendix functions as a safe-house for beneficial bacteria, and confirm fecal communities as fairly reliable proxies for consortia along the lower gut. We encourage others to make similar use of natural or accidental losses for probing the primate gut microbiome.

Water hemlock poisoning in cattle: Ingestion of immature Cicuta maculata seed as the probable cause.

It is well understood that water hemlock tubers are highly toxic to animals and to humans. However, this is the first time that immature seed from (Cicuta maculata) has been implicated in livestock poisoning. Nine mature Hereford cows from a herd of 81 died in northwestern Utah after ingesting immature seed heads of water hemlock (Cicuta maculata) in late summer. No obvious signs of poisoning were reported as all nine were found dead near the banks of the stream where water hemlock was found. Upon discovery of the dead cows, the remaining 72 cows were immediately removed from the pasture and no further losses occurred. Field necropsy of 3 of the dead cows and follow-up serology and histopathological examination of selected tissues did not identify any bacterial or viral causes. History of ingestion of large quantities of water hemlock seed, the acute nature of the deaths, chemical comparison of seed with toxic tubers and follow-up mouse bioassay testing supported the diagnosis of water hemlock poisoning. Seed heads collected from the neighboring pasture upstream and across the fence from the poisoned cattle and tubers collected from grazed plants were chemically analyzed and found to contain cicutoxin, and high levels of two cicutol-like derivatives (cicutol-#1 and #2) as well as other unidentified polyacetylene compounds. Seeds and tubers from suspected plants were semi-quantified and compared to archive samples of highly toxic tubers used in previous experiments. The immature hemlock seed contained less cicutoxin (0.01 times), but 9.5 and 22.5 times more cicutol-#1 and cicutol-#2 respectively, compared to the archive sample. Tubers from the grazed plants contained 4.6 times more cicutoxin and 9.8 and 18.8 times more cicutol-#1 and cicutol-#2 respectively, compared to the archive sample. Mouse bioassays with water extracts of immature seed and tubers from grazed plants demonstrated both were highly toxic and of greater toxicity when compared to archived sample.

Poisoning due to water hemlock.

INTRODUCTION: Water hemlock, which encompasses a range of species divided across two genera (Cicuta and Oenanthe), are regarded as being among the most poisonous plants both in North America and in the United Kingdom. Despite their toxicity, the literature consists almost entirely of case reports. AIM: The aim of this review is to summarize this literature by covering all aspects of taxonomy and botanical characterization, principal toxins, basic pharmacology including mechanisms of toxicity, and the clinical features, diagnosis, and management of poisoning. MECHANISMS OF TOXICITY: The principal toxins, cicutoxin and oenanthotoxin, belong to a group of C17 conjugated polyacetylenes. They act as (noncompetitive) gamma-aminobutyric acid antagonists in the central nervous system (CNS), resulting in unabated neuronal depolarization that can lead to seizures. Ingestion of even a small amount of plant matter may result in severe intoxication. FEATURES: After ingestion, the patient is most likely to experience CNS stimulatory effects including seizures that, in the absence of aggressive supportive care, can result in death. Other features include nausea, vomiting, diarrhea, tachycardia, mydriasis, rhabdomyolysis, renal failure, coma, respiratory impairment, and cardiac dysrhythmias. MANAGEMENT: Treatment consists mainly of prompt airway management and seizure control, plus decontamination if achieved early and after stabilization. In the event of renal failure, the use of hemodialysis has been employed successfully. CONCLUSIONS: The ingestion of water hemlock can lead to serious complications that may be fatal. Prognosis is good, however, if prompt supportive care is provided.

Polyacetylenes from sardinian Oenanthe fistulosa: a molecular clue to risus sardonicus.

An investigation of Oenanthe fistulosa from Sardinia afforded oenanthotoxin (1a) and dihydrooenanthotoxin (1b) from the roots and the diacetylenic epoxydiol 2 from the seeds. The absolute configuration of 1a and 1b was established as R by the modified Mosher's method, and the structure of 2 by chemical correlation with (+)-(3R,8S)-falcarindiol. Oenanthotoxin (1a) and dihydrooenanthotoxin (1b) were found to potently block GABAergic responses, providing a molecular rationale for the symptoms of poisoning from water-dropwort (Oenanthe crocata) and related plants. These observations bear relevance for a series of historical and ethnopharmacological observations on the identification of the Sardonic herb and the molecular details of the facial muscular contraction caused by its ingestion (risus sardonicus).

Application of liquid chromatography-mass spectrometry to the investigation of poisoning by Oenanthe crocata.

Liquid chromatography-mass spectrometry (LC-MS) analysis of methanol extracts of Oenanthe crocata roots revealed that oenanthotoxin co-eluted with another major polyalkyne, 2,3-dihydro-oenanthotoxin, using the existing high performance liquid chromatography (HPLC) method (isocratic elution from C18 with aqueous methanol) for investigating Oenanthe poisoning. Positive ES or APCI gave [(M+H)-H(2)O](+) and its methanol adduct as major ion species for oenanthotoxin, whereas 2,3-dihydro-oenanthotoxin formed [M+H](+) and its methanol adduct. The two polyalkynes could be chromatographically resolved on C18 by gradient elution with aqueous acetonitrile. This provides superior analysis for oenanthotoxin using HPLC with photodiode array (PDA) detection alone, but for LC-MS/MS aqueous acetonitrile was less suitable due to poor ionisation and, with APCI, an increase in the relative abundance of a [M-1](+) species, which could confuse compound assignment. HPLC-PDA and LC-MS/MS methods using an aqueous acetonitrile or aqueous methanol mobile phase, respectively, were successful when applied to the analysis of the stomach contents of a pony suspected to have eaten O. crocata. Relevant product ion spectra, by ion trap MS/MS, accurate mass data and complete sets of (1)H and (13)C NMR spectral assignments are given for the two compounds.

Clinical spectrum of accidental hemlock poisoning: neurotoxic manifestations, rhabdomyolysis and acute tubular necrosis.

In the past, hemlock poisoning was only known for its neurotoxic effects; quite recently non-neurological features, consisting of rhabdomyolysis and acute renal failure, have been also described. Here we report our experience with these clinical findings, which we frequently observe in accidental hemlock poisoning. Between 1972 and 1990 we studied 18 patients: 17 of them were poisoned by conline (an alkaloid of Conium maculatim) in Apulia (Italy), and one by cicutoxin (the active principle of water hemlock) in New Mexico (USA). In the non-rapidly-fatal cases we tested myoglobinuria, serum muscle enzymes, and renal function. In the patients with acute renal failure we performed microscopical examination of kidney specimens; immunohistochemistry was carried out to identify myoglobin and actin in tubules. Coniine was detected in urine, serum, or tissues. Neurological features were present in all of our cases: coniine had a curare-like effect on the neuromuscular junction, whereas cicutoxin was convulsant on the central nervous system. In addition rhabdomyolysis was noted in the 17 subjects poisoned by coniine. Acute renal failure was observed in five patients; it was confirmed by histological evidence of tubular necrosis with intratubular deposition of myoglobin and actin released by rhabdomyolysis. Our cases seem to be the first with histopathologically proven acute tubular necrosis in coniine intoxication. In conclusion, in hemlock poisoning neurotoxic manifestations may be accompanied by rhabdomyolysis and acute tubular necrosis; increased awareness of these clinical features is recommended in order to improve the diagnostic and therapeutic procedure.

Hemlock water dropwort poisoning.

Severe plant poisoning is relatively uncommon in adults. We report two adults who ingested hemlock water dropwort roots, having mistaken them for wild parsnip. One developed prolonged convulsions, severe metabolic acidosis and respiratory distress requiring mechanical ventilation. The toxin--oenanthotoxin--was detected in the gastric aspirate and measured by high performance liquid chromatography.

Cicutoxin poisoning (water hemlock).

A 54-year-old man had a severe case of cicutoxin poisoning following the ingestion of water hemlock (Cicuta maculate). The administration of thiopentone sodium by continuous intravenous infusion proved the optimum method of controlling the gross and irregular body movements and of managing the metabolic acidosis and the cholinergic crisis characteristic of this type of poisoning. This report includes a detailed neurologic and biochemical assessment of the condition.