Evaluation of the toxicological effects of Neltuma alpataco (Prosopis alpataco) pod alkaloid extract.

The pods of Neltuma spp. have shown potential as a source of protein and energy in livestock. However, prolonged consumption of some of these species can lead to neurological symptoms in ruminants. This study aimed to determine the alkaloid content, as well as the in vitro and in vivo effects of an alkaloid-enriched extract (AEE) from N. alpataco pods. High performance liquid chromatography-high resolution mass spectrometry (HPLC-HRMS) identified juliprosine and juliprosopine as primary alkaloids, with juliprosine being most abundant. AEE from N. alpataco demonstrated dose-dependent cytotoxicity on glioma cells after 48 h, with a 50% cytotoxic concentration (CC50) of 24.69 µg/mL. However, the release of LDH was observed only at the highest tested concentration, indicating cellular damage. Further examination through phase-contrast microscopy and dual acridine orange/ethidium bromide fluorescence staining revealed morphological changes consistent with an apoptotic mechanism of cell death, ultimately leading to secondary necrosis. Finally, the LD50 after intraperitoneal injection in mice was determined to be 12.98 mg/kg. Taken together, these findings demonstrated for the first time the in vivo and in vitro toxicity of the AEE from N. alpataco pods.

Hepatic encephalopathy in swine experimentally poisoned with Senna occidentalis seeds: Effects on astrocytes.

Senna occidentalis may be accidently ingested by humans and animals. In this study, the percentages of S. occidentalis seeds necessary for experimental reproduction of hepatic encephalopathy were determined in a pig model and the biochemical and microscopic pathology is described in detail, with emphasis on the astrocytes. The experimental groups (G1, G2 and G3) were fed rations containing 5%, 7.5% and 10% of S. occidentalis seeds for 7-11 days. Pigs from the three experimental groups showed incoordination, ataxia, disorientation, head pressing, anorexia, recumbency and depression. In addition, the enzymes aspartate aminotransferase, alkaline phosphatase and creatine phosphokinase increased in all treated animals, which also showed higher serum total bilirubin and ammonia levels than in the control group (C). Microscopically, all experimental animals revealed acute hepatocellular swelling, multifocal coagulative necrosis in the pancreas, necrosis in the cardiac muscle, severe spongiosis in brain white and grey matter, and Alzheimer type II astrocytes in grey matter of the cerebral cortex. These cells were glial fibrillary acidic protein (GFAP) negative in G3. In white matter, a decrease in the positive area occupied by GFAP-immunolabelling and in the number of astrocytes per immunoreactive area was observed in G3 animals (5.35 ± 1.14% and 410 ± 45 cells/mm2, respectively) compared to the C animals (13.93 ± 1.59% and 581 ± 36 cells/mm2, respectively). This loss of GFAP was accompanied by alterations in astrocyte morphology, such as shrinkage of the cell body and retraction of the extending processes. This pig model of ammonia-mediated astrocyte damage could be used to study not only poisoning by S. occidentalis, but also other medical conditions resulting in hepatoencephalopathy.