Arthropod and Wolbachia Identification – Kudzu Bug

December 4, 2025

Sample information
Picture
Location
Collection date 09/26/2025
Captive / Cultivated? Wild-caught
Group NCSSM
Observations
Using mason jars, the kudzu bugs were caught from the kudzu patch near Western Piedmont Community College. A leaf was plucked from the kudzu, which contained a group of kudzu bugs on its backside. All the insects used in the experiment were caught on the same day, in the afternoon.
Putative identification Arthropoda Insecta Hemiptera Plataspidae Megacopta Megacopta cribraria

Methods
Extraction kit DNeasy (Qiagen) blood and tissue kit
DNA extraction location Partial abdomen
Single or Duplex PCR Single Reaction
Gel electrophoresis system MiniPCR
Buffer TBE
DNA stain Cyber Green
Gel images
Protocol notes
DNA Extraction: The kudzu bug was first thoroughly washed with molecular biology-grade water, and then its abdomen was separated from the rest of the body using a sterile pair of tweezers, scalpel, and Petri dish. The reproductive organs were then transferred into a centrifuge tube containing ATL buffer and ground thoroughly with a pestle afterward.

Gel Electrophoresis Lanes:
Red Box: Sample Analyzed
Lane 8: Sample Amplified with Arthropod Primers
Lane 9: Sample Amplified with Wolbachia Primers
Lane 10: Positive Wolbachia Control
Lane 11: Negative Wolbachia Control
Lane 12: Positive Arthropod Control
Lane 13: Negative Arthropod Control

Results
Wolbachia presence Yes
Confidence level Medium
Explanation of confidence level

For this kudzu bug sample, the bands indicative of both Arthropod (~708 bp) and Wolbachia (~438 bp) presence were observed during gel electrophoresis. The confidence level was deemed medium, since the Positive Wolbachia Control sample result was incorrect.
Wolbachia 16S sequence
Arthropod COI sequence
Summary The Megacopta cribraria was found to be postive for Wolbachia.
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