Development of a PCR-based method for detection of Delphinium species in poisoned cattle

Authors: Cook, Daniel; Pfister, James; Constantino, John; Roper, Jessie; Gardner, Dale; Welch, Kevin; HAMMOND, ZACHARY - Northeastern University; Green, Benedict - Ben

Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/8/2015
Publication Date: 1/8/2015
Publication URL: https://handle.nal.usda.gov/10113/62820
Citation: Cook, D., Pfister, J.A., Constantino, J., Roper, J.M., Gardner, D.R., Welch, K.D., Hammond, Z.J., Green, B.T. 2015. Development of a PCR-based method for detection of Delphinium species in poisoned cattle. Journal of Agricultural and Food Chemistry. 63:1120-1125.

Interpretive Summary: Toxic plants 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 these experiments 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 primers specific to 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.

Technical Abstract: Toxic plants 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 these experiments 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 diagnoise larkspur and perhaps other toxic plant-caused losses.