Salivary AsHPX12 influences pre-blood meal associated behavioral properties in Anopheles stephensi

Background & objectives: A successful blood meal acquisition process by an adult female mosquito is accomplished through salivary glands, which releases a cocktail of proteins to counteract the vertebrate host’s immune homeostasis. Here, we characterize a salivary-specific Heme peroxidase family member HPX12, originally identified from Plasmodium vivax infected salivary RNAseq data of the mosquito Anopheles stephensi. Methods: To demonstrate we utilized a comprehensive in silico and functional genomics approach. Results: Our dsRNA-mediated silencing experiments demonstrate that salivary AsHPX12 may regulate pre-blood meal-associated behavioral properties such as probing time, probing propensity, and host attraction. Altered expression of the salivary secretory and antennal proteins expression may have accounted for salivary homeostasis disruption resulting in the unusual fast release of salivary cocktail proteins and delayed acquisition of blood meal in the AsHPX12 knockdown mosquitoes. We also observed a significant parallel transcriptional modulation in response to blood feeding and P. vivax infection. Interpretation & conclusion: With this work, we establish a possible functional correlation of AsHPX12 role in the maintenance of salivary physiological-homeostasis, and Plasmodium sporozoites survival/ transmission, though the mechanism is yet to unravel.

Entering and exiting behaviour of the phlebotomine sand fly Lutzomyia longiflocosa (Diptera: Psychodidae) in rural houses of the sub-Andean region of Colombia

The present study identified the entering and exiting sites for Lutzomyia longiflocosa in rural houses of the sub-Andean region in Colombia. Entering sites were identified with sticky traps set up outside the bedrooms, around the eave openings, and with cage traps enclosing the slits in the doors and windows inside the bedrooms. Exiting sites were identified by releasing groups of females indoors. These females were blood fed and marked with fluorescent powders. Females were recaptured with the trap placement described above but set up on the opposite sides of the openings. In the entering experiment, a significantly higher number of females were captured in the sticky traps at the zone nearest the eave openings (n = 142) than those captured in the other zones of the trap (n = 52); similarly, a higher number of females were captured on the front side of the house (n = 105) than at the rear side (n = 37). Only two females were collected in the cage trap. In the exiting experiment, at the ceiling, the highest percentage (86.2%) of females was recaptured with sticky traps nearest the eave openings and on the front side of the house (70.0%). Seven females were collected in the cage trap. Lu. longiflocosa entered and exited houses primarily through the eave openings in a non-random pattern in relation to the sides of the house.

Fabrication of an olfactometer for mosquito behavioural studies.

Background & objectives: Olfaction is the major sensory modality involved in the resource searching behaviour of insects including vector mosquitoes (Diptera: Culicidae). To date, our current countrywide knowledge on the host-seeking behaviour of Iranian mosquitoes is mainly confined to host preference which has exclusively come from field studies. Olfactometer is a scientific tool by which more naive aspects of man-vector contact can be clarified under controlled and less biased conditions. Methods: The wind tunnel and stimulus delivery system was constructed from acrylic materials based on previously introduced models with some modifications. Air supply and required light were ensured by a powerful compressor and incandescent bulbs, respectively. Desired level of temperature was maintained by controllable heating radiators. For humidity production a unique in-built piezo system was devised in the course of the air flow. Fine regulators facilitated the continuous generatation of the humidity at a preset level. Results: Titanium tetrachloride smoke plus monitoring of the wind speed revealed that the flow of air was proper and invariable. A desired level of humidity and temperature could be set up in just 10 and 15–45 min, respectively. These physical parameters varied only ±2% (humidity) and ±0.15ºC (temperature) in a typical 20 min duration. Conclusion: The first sophisticated olfactometer in the field of medical entomology in Iran is reported here. Fast set up and stability of physical parameters are its salient features. It is expected that with the aid of this olfactometer further information on the physiological principles of the host-seeking behaviour of mosquitoes become available soon.