Spatial Distribution of PCR-Identified Species of Anopheles gambiae senu lato (Diptera: Culicidae) Across Three Eco-Vegetational Zones in Cross River State, Nigeria.

Anopheles gambiae sensu lato complex (An. gambiae s.l.) describes a group of nine morphologically indistinguishable members that vary in their distribution, ability to transmit malaria, and susceptibility to pyrethroids. Here, we recorded the spatial patterns of PCR-identified An. gambiae s.l. complex species collected from four sites in Cross River State, Nigeria that represented three different ecological zones. Trapping was conducted between October 2015 and June 2016. Anopheles gambiae s.l. complex species identification was performed using species-specific primers followed by An. gambiae and An. coluzzii differentiation using the restriction fragment length polymorphism (RFLP) method. Bivariate and multivariate logistic regression models were used to identify ecological and seasonal variables closely associated with An. coluzzii and An. gambiae distribution. Out of 1,388 An. gambiae s.l. successfully amplified, 1,074 (77.4%) were An. coluzzii, 278 (20%) were An. gambiae, and 25 (1.8%) were hybrids (An. coluzzii/An. gambiae). A very small number of An. arabiensis (0.8%, n = 11) were also collected. Statistical analysis indicated that An. coluzzii is predominant in Guinea-savannah and tropical rainforest, and is highly associated with rainy seasons, while, An. gambiae is prevalent in mangrove swamp forest during dry seasons. Only 13 An. gambiae s.l. females were infected with Plasmodium falciparum (P. falciparum). The sporozoite infection rate was higher in mangrove swamp forest (53.8%, n = 7) than in rain forest (38.5%, n = 5) followed by Guinea-savannah (7.7%, n = 1) ecological zones. These results provide important insights for strategic planning of malaria control programs in Nigeria.

New Records and Updated Checklist of Phlebotomine Sand Flies (Diptera: Psychodidae) From Liberia.

Phlebotomine sand flies from three counties in Liberia were collected from January 2011 to July 2013. In total, 3,118 sand flies were collected: 18 species were identified, 13 of which represented new records for Liberia. An updated taxonomic checklist is provided with a brief note on sand fly biology, and the disease vector potential for species is discussed.

Pyrethroid Insecticide Resistance Mechanisms in the Adult Phlebotomus papatasi (Diptera: Psychodidae).

Phlebotomus papatasi is one of the most medically important sand fly species in the Old World, serving as a vector of Leishmania parasites and phleboviruses. Chemical control is still considered the most effective method for rapidly reducing populations of flying insects involved in vector-borne disease transmission, but is increasingly threatened by insecticide resistance in the target insect posing significant problems for entomologists responsible for control programs. This study was conducted to determine pyrethroid resistance mechanisms and the biological, physiological, and molecular impacts of resistance in Ph. papatasi, and to compare their resistance mechanisms against those reported for mosquitoes and other intensely studied dipterans. Field-collected Ph. papatasi from Aswan, Egypt, were subjected to sublethal doses of permethrin and reared as a resistant strain under laboratory conditions through 16 generations. Biological parameter observations of resistant Ph. papatasi revealed an association of resistance with productivity cost. Physiological analysis revealed that concentrations of oxidase and esterase enzymes increased in early generations of the resistant colony, and then subsided through the F16 generation to levels similar to those in a susceptible colony. The activity levels of acetylcholinesterase were higher in field-collected Ph. papatasi than in susceptible colony flies, but decreased significantly despite subsequent exposure to permethrin. The molecular search for gene mutations in the resistant strain of Ph. papatasi failed to identify any mutations common in pyrethroid-resistant mosquitoes. Our study revealed that the mechanism of pyrethroid resistance in sand flies is different than that in mosquitoes, at least at the genetic level.

The Seasonality and Ecology of the Anopheles gambiae complex (Dipetra: Culicidae) in Liberia Using Molecular Identification.

Members of the Anopheles gambiae sensu lato (Giles) complex define a group of seven morphologically indistinguishable species, including the principal malaria vectors in Sub-Saharan Africa. Members of this complex differ in behavior and ability to transmit malaria; hence, precise identification of member species is critical to monitoring and evaluating malaria threat levels. We collected mosquitoes from five counties in Liberia every other month from May 2011 until May 2012, using various trapping techniques. A. gambiae complex members were identified using molecular techniques based on differences in the ribosomal DNA (rDNA) region between species and the molecular forms (S and M) of A. gambiae sensu stricto (s.s) specimens. In total, 1,696 A. gambiae mosquitoes were collected and identified. DNA was extracted from legs of each specimen with species identification determined by multiplex polymerase chain reaction using specific primers. The molecular forms (M or S) of A. gambiae s.s were determined by restriction fragment length polymorphism. Bivariate and multivariate logistic regression models identified environmental variables associated with genomic differentiation. Our results indicate widespread occurrence of A. gambiae s.s., the principal malaria vector in the complex, although two Anopheles melas Theobald/A. merus Donitz mosquitoes were detected. We found 72.6, 25.5, and 1.9% of A. gambiae s.s specimens were S, M, and hybrid forms, respectively. Statistical analysis indicates that the S form was more likely to be found in rural areas during rainy seasons and indoor catchments. This information will enhance vector control efforts in Liberia.

Field responses of Anopheles gambiae complex (Diptera: Culicidae) in Liberia using yeast-generated carbon dioxide and synthetic lure-baited light traps.

Malaria infection is a serious public health problem throughout Liberia, but vector surveillance is limited or nonexistent in remote regions of the country. To better understand the spatial and temporal distribution of malaria vectors in Liberia and to support vector and malaria activities of the Liberian Ministry of Health, a study was conducted to determine the efficacy of light traps baited with a synthetic lure and CO2 for capturing Anopheles gambiae sensu lato (Giles). Traps with a ultraviolet, light-emitting diode, and incandescent lights baited with a synthetic skin lure and CO2 combinations were evaluated at four field sites in three counties of Liberia for five consecutive nights every 8 wk during 2011. In total, 4,788 mosquitoes representing 56 species from nine genera were collected throughout the 30-wk study; An. gambiae s. l. comprised 32% and of the148 An. gambiae s. s. collected, 85% were of the S form. A greater percentage of An. gambiae s. l. were collected in ultraviolet traps baited with a synthetic lure and CO2 compared with any other trap configuration. The influence of trap configuration on conclusions from surveillance efforts, specifically with regards to An. gambiae is discussed.

Behavioral and physiological response of Musca domestica to colored visual targets.

A better understanding of the visual attraction of house flies to colors and patterns is needed to improve fly trap performance. This study combined physiological responses measured with electroretinogram studies of the house fly's compound eyes and ocelli with behavioral attraction of flies to reflective colors and patterns in light tunnel assays. Compound eye and ocellar electroretinogram responses to reflected light were similar, with the largest responses to white and blue followed by yellow, red, green, and black. However, data from light tunnel behavioral assays showed that flies were attracted to white and blue light but were repelled by yellow. The addition of a black line pattern enhanced the attractiveness of blue visual targets, whereas yellow lines decreased attractiveness. Sensory input from the compound eye and the ocellus seems to be integrated to direct fly behavior. There is a direct correlation of house fly attractiveness to visual targets and the intensity of electrophysiological response, except for the yellow targets, which repel flies despite of intense electrophysiological response.

A case study of Megaselia scalaris (Diptera: Phoridae) causing ocular myiasis in a western hognose snake.

Late instar larvae of the scuttle fly, Megaselia scalaris Loew, were found near the right eye of a live captive-reared western hognose snake, Heterodon nasicus Baird and Girard. Dissection and removal of the snake's dorsal cranial bones revealed tissue degradation of the infected eye, the optic nerve, and the brain case; we suggest that these factors contributed to the death of this snake. This case study further demonstrates the opportunistic behavior of M. scalaris.