Nepenthes pitcher plants produce modified jug-shaped leaves to attract, trap and digest insect prey. We used 16S rDNA cloning and sequencing to compare bacterial communities in pitcher fluids of each of three species, namely Nepenthes ampullaria, Nepenthes gracilis and Nepenthes mirabilis, growing in the wild. In contrast to previous greenhouse-based studies, we found that both opened and unopened pitchers harbored bacterial DNA. Pitchers of N. mirabilis had higher bacterial diversity as compared to other Nepenthes species. The composition of the bacterial communities could be different between pitcher types for N. mirabilis (ANOSIM: R = 0.340, p < 0.05). Other Nepenthes species had similar bacterial composition between pitcher types. SIMPER showed that more than 50 % of the bacterial taxa identified from the open pitchers of N. mirabilis were not found in other groups. Our study suggests that bacteria in N. mirabilis are divided into native and nonnative groups.

Nepenthes pitcher plants are colonized by a variety of specialized arthropods. As Aedes mosquitoes are container breeders, Nepenthes pitchers are a potential candidate oviposition site for vector species, such as Aedes aegypti (L.) and Aedes albopictus (Skuse). However, Aedes spp. are not commonly encountered in Nepenthes pitchers, and the environment inside the pitchers of some species is lethal to them. One exception is Nepenthes ampullaria Jack, whose pitchers are known to be colonized by Ae. albopictus on very rare occasions. Given that Ae. albopictus larvae can survive in N. ampullaria pitcher fluids, we sought to determine why pitcher colonization is rare, testing the hypothesis that gravid Aedes mosquitoes are deterred from ovipositing into container habitats that have similar characteristics to N. ampullaria pitchers. Using plastic ovitraps of different sizes, colors, and with different types of fluids (based on the characteristics of N. ampullaria pitchers), we compared oviposition rates by Aedes mosquitoes in urban and rural areas within the geographical range of N. ampullaria near Kuala Lumpur, Malaysia. Ovitraps that were black and large (>250-ml capacity) accumulated significantly more eggs than ovitraps that were smaller, or green in color. In terms of size and color, small, green ovitraps are analogous to N. ampullaria pitchers, indicating that these pitchers are not particularly attractive to gravid Ae. albopictus. Although Aedes spp. are capable of colonizing N. ampullaria pitchers, the pitchers are relatively unattractive to gravid females and do not represent a significant habitat for larvae of dengue vectors at present.

1. Nepenthes pitcher plants produce fluid-containing animal traps that are colonised by a variety of specialised arthropods, especially dipterans. However, container-breeding vector mosquitoes, such as Aedes albopictus Skuse have rarely been recorded from pitchers. Increasing overlap in the geographical ranges of Nepenthes and Ae. albopictus in urban parts of Southeast Asia owing to urbanisation highlights a growing need to investigate the potential role of pitchers as larval habitats for vector mosquitoes.2. The ability of Ae. albopictus larvae to survive in three common lowland Nepenthes in Peninsular Malaysia that are most likely to co-occur with Ae. albopictus [viz., Nepenthes ampullaria Jack, Nepenthes gracilis Korth., and Nepenthes mirabilis (Lour.) Druce] was investigated.3. The larval survival rates of Ae. albopictus in pitcher fluids of the three Nepenthes species were determined, then the effects of low pH, larvicidal agents (such as microbes, predators, and chemical compounds) through manipulative experiments were investigated.4. It was found that pitchers represent a hostile environment to Ae. albopictus, but that the principal cause of larval mortality varies among Nepenthes species (i.e. low fluid pH in N. gracilis, predation by Toxorhynchites acaudatus Leicester larvae in N. ampullaria, and microbial activity in N. mirabilis). It was concluded that Nepenthes pitchers are generally not suitable larval habitats for Ae. albopictus. However, the pitcher environment of N. ampullaria is worthy of further study, as pitchers that lack predators are nevertheless rarely colonised by Ae. albopictus, indicating that other aspects of the host pitcher environment inhibit oviposition or larval survivorship.

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