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Development of a PCR-Based Method for Detection of Delphinium Species in Poisoned Cattle

Daniel Cook, James A. Pfister, John R. Constantino, Jessie M. Roper, Dale R. Gardner, Kevin D. Welch, Zachary J. Hammond, Benedict T. Green
Journal of agricultural and food chemistry 2015 v.63 no.4 pp. 1220-1225
DNA primers, toxicity, Delphinium occidentale, analytical chemistry, cattle, death, detection, diagnostic techniques, digestion, ingestion, microscopy, poisonous plants, polymerase chain reaction, rumen
Toxic plants such as Delphinium spp. (i.e., larkspur) are a significant cause of livestock losses worldwide. Correctly determining the causative agent responsible for the death of an animal, whether by disease, poisonous plant, or other means, is critical in developing strategies to prevent future losses. The objective of this study was to develop an alternative diagnostic tool to microscopy and analytical chemistry to determine whether a particular poisonous plant was ingested. Polymerase chain reaction (PCR) is a tool that may allow detection of the genetic material from a specific plant within a complex matrix such as rumen contents. A pair of oligonucleotide primers specific to Delphinium spp. (i.e., larkspur) was developed; using these primers, a PCR product was detected in samples from an in vivo, in vitro, and in vivo/in vitro coupled digestion of Delphinium occidentale. Lastly, larkspur was detected in a matrix of ruminal material where the amount of larkspur was far less than what one would expect to find in the rumen contents of a poisoned animal. The PCR-based technique holds promise to diagnose larkspur and perhaps other toxic plant caused losses.