DETERMINATION OF ANOPHELES GAMBIAE LARVAL DNA IN THE GUT OF INSECTIVOROUS DRAGONFLY (LIBELLULIDAE) NYMPHS BY POLYMERASE CHAIN REACTION

Type

Article

Date of Publication

2003

Original Publication

Journal of the American Mosquito Control Association

Volume

19

Series / Issue

Issue: 2

Pages

163-165

Contributed by

American Mosquito Control Association

Notes

Author(s) - MARIA E. MORALES, DAWN M. WESSON, IAN W. SUTHERLAND, DANIEL E. IMPOINVIL, CHARLES M. MBOGO, JOHN I. GITHURES, JOHN C. BEIER

 

Abstract

We examined the predator-prey relationship between larvae of the malaria mosqtito Anophelesgambiae and nymphs of the dragonfly (Libellulidae). Studies were conducted to determine whether polymerasechain reaction (PCR) can be used to detect DNA of Az. gambiae in the gut of libellulid nymphs, and to determinehow long after feeding on An. gambiae that mosquito DNA remains detectable by PCR. Total DNA was exhactedfrom the gut contents of libellulid nymphs by using 2 types of DNA extraction methods. The target sequencefor the diagnostic PCR was the intergenic spacer regions of the ribosomal DNA gene locus. These sequenceswere analyzed by using An. gambiae complex-specific primers. After analyzing nymphal gut contents with PCRat regular postfeed intervals, a 390-base pair product could be amplified. The presence of mosquito larvae wasvisually confirmed for up to 40 min after feeding. Regardless of the number of mosquito larvae ingested,libellulid gut contents could be amplified or visually seen up to I h of digestion. This result indicates the nymphshave a high rate of digestion and that PCR with An. gambiae comp'lex primers will be best utilized within I hafter feeding as a detection system. This study confirmed that dragonfly nymphs feed well on anopheline larvae,and that mosquito DNA, although rapidly digested, can be successfully recovered and detected from withinnymphal digestive tracts.

Notes

Author(s) - MARIA E. MORALES, DAWN M. WESSON, IAN W. SUTHERLAND, DANIEL E. IMPOINVIL, CHARLES M. MBOGO, JOHN I. GITHURES, JOHN C. BEIER