A temporal comparison of sex-aggregation pheromone gland content and dynamics of release in three members of the Lutzomyia longipalpis (Diptera: Psychodidae) species complex

摘要

Background:udLutzomyia longipalpis is the South American vector of Leishmania infantum, the etiologic agent of visceral leishmaniasis (VL). Male L. longipalpis produce a sex-aggregation pheromone that is critical in mating, yet very little is known about its accumulation over time or factors involved in release. This laboratory study aimed to compare accumulation of pheromone over time and determine factors that might influence release in three members of the L. longipalpis species complex.ududMethodology/Principal findings:udWe investigated male sex-aggregation pheromone gland content at different ages and the release rate of pheromone in the presence or absence of females under different light conditions by gas chromatography-mass spectrometry (GC-MS). Pheromone gland content was determined by extraction of whole males and pheromone release rate was determined by collection of headspace volatiles. Pheromone gland content appeared age-related and pheromone began to accumulate between 6 to 12 h post eclosion and gradually increased until males were 7–9 days old. The greatest amount was detected in 9-day old Campo Grande males ((S)-9-methylgermacrene-B; X ± SE: 203.5 ± 57.4 ng/male) followed by Sobral 2S males (diterpene; 199.9 ± 34.3) and Jacobina males ((1S,3S,7R)-3-methyl-α-himachalene; 128.8 ± 30.3) at 7 days old. Pheromone release was not continuous over time. During a 4-hour period, the greatest quantities of pheromone were released during the first hour, when wing beating activity was most intense. It was then substantially diminished for the remainder of the time. During a 24 h period, 4–5 day old male sand flies released approximately 63 ± 11% of the pheromone content of their glands, depending on the chemotype. The presence of females significantly increased pheromone release rate. The light regime under which the sand flies were held had little influence on pheromone release except on Sobral 2S chemotype.ududConclusions/Significance:udAccumulation of pheromone appears to occur at different rates in the different chemotypes examined and results in differing amounts being present in glands over time. Release of accumulated pheromone is not passive, but depends on biotic (presence of females) and abiotic (light) circumstances. There are marked differences in content and release between the members of the complex suggesting important behavioural, biosynthetic and ecological differences between them.ududAuthor summary:udThe Dipteran subfamily Phlebotominae includes the genera Lutzomyia and Phlebotomus among which several species are important vectors of parasitic and bacterial pathogens. The sand fly Lutzomyia longipalpis is considered the main vector of visceral leishmaniasis (VL) in the New World. Based on the main component of the male sex-aggregation pheromone gland, different sex pheromone-producing populations (chemotypes) of L. longipalpis are recognized in Brazil. Given the importance of the sex-aggregation pheromones in the biology of this species complex, we present here the first attempt to study how pheromone accumulates in the glands over time and factors that might influence its release in the three most common chemotypes from Brazil. Our results demonstrated that pheromone first starts to accumulate a few hours post-eclosion (6–12 h) and this continues over 15 days. Pheromone release is a dynamic process which varies between the 3 chemotypes depending on biotic factors, such as light regime and presence/absence of conspecific females. This work provides valuable information, critical to our understanding of the behaviour and ecology of L. longipalpis sand flies and which will contribute to investigations to improve field-based pheromone control and monitoring of L. longipalpis sand flies.