Determination of Anopheles gambiae larval DNA in the gut of insectivorous dragonfly (Libellulidae) nymphs by polymerase chain reaction.
We examined the predator-prey relationship between larvae of the malaria mosquito Anopheles gambiae and nymphs of the dragonfly (Libellulidae). Studies were conducted to determine whether polymerase chain reaction (PCR) can be used to detect DNA of An. gambiae in the gut of libellulid nymphs, and to determine how long after feeding on An. gambiae that mosquito DNA remains detectable by PCR. Total DNA was extracted from the gut contents of libellulid nymphs by using 2 types of DNA extraction methods. The target sequence for the diagnostic PCR was the intergenic spacer regions of the ribosomal DNA gene locus. These sequences were analyzed by using An. gambiae complex-specific primers. After analyzing nymphal gut contents with PCR at regular postfeed intervals, a 390-base pair product could be amplified. The presence of mosquito larvae was visually 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 1 h of digestion. This result indicates the nymphs have a high rate of digestion and that PCR with An. gambiae complex primers will be best utilized within 1 h after 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 within nymphal digestive tracts.
Morales
Maria E
ME
Department of Tropical Medicine, 1430 Tulane Avenue, University School of Public Health and Tropical Medicine, Tulane University, New Orleans, LA 70112, USA.
Wesson
Dawn M
DM
Sutherland
Ian W
IW
Impoinvil
Daniel E
DE
Mbogo
Charles M
CM
Githure
John I
JI
Beier
John C
JC
D000818:Animals; D000852:Anopheles / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D020387:Food Chain; D007313:Insecta / Q000502:physiology*; D007814:Larva; D009758:Nymph / Q000502:physiology; D016133:Polymerase Chain Reaction
D017931:DNA Primers