Resting Behavior of Malaria Vectors in Ghana and Its Implication on Vector Control.

Background: In Sub-Saharan Africa, there is widespread use of long-lasting insecticidal nets (LLINs) and Indoor residual spraying (IRS) to help control the density of malaria vectors and decrease the incidence of malaria in communities. An understanding of the interactions between increased insecticide use and resting behaviour patterns of malaria mosquitoes is important for an effective vector control programme. This study was carried out to investigate the resting behavior, host preference and infection with Plasmodium falciparum of malaria vectors in Ghana in the context of increasing insecticide resistance in malaria vectors in sub-saharan Africa.Methods: Indoor and outdoor resting Anopheline mosquitoes were sampled during the dry and rainy seasons in five sites that were in 3 ecological landscapes [Sahel savannah (Kpalsogou, Pagaza, Libga), Coastal savannah (Anyakpor) and Forest (Konongo) zones] using pyrethrum spray catches (PSC), mechanical aspiration (Prokopack) for indoor collections, pit shelter and Prokopack for outdoor collections. PCR based molecular diagnostics were used to determine mosquito speciation, genotype for knockdown resistance mutations (L1014S and L1014F), G119S Ace-1 mutation, specific host blood meal origins and sporozoite infection in field collected mosquitoes.Results: Anopheles gambiae s. l. was the predominant species (89.95%, n = 1,718), followed by An. rufipes (8.48%, n=162), and An. funestus s. l. (1.57%, n = 30). Sibling species of the Anopheles gambiae revealed An. coluzzii accounted for 63% (95% CI: 57.10 – 68.91), followed by An. gambiae s. s [27% (95% CI: 21.66 – 32.55)], and An. arabiensis [9% (95% CI: 6.22 – 13.57)]. The mean resting density of An. gambiae s. l. was higher outdoors (79.63%; 1,368/1,718) than indoors (20.37%; 350/1,718) (z = -4.815, p< 0.0001). The kdr west L1014F and the Ace-1 mutations were highest in indoor resting An. coluzzii and An. gambiae in the sahel-savannah sites compared to the forest and coastal savannah sites. Overall, the blood meal analyses revealed a large proportion of the malaria vectors preferred feeding on humans (70.2 %) than animals (29.8%) in all sites. The sporozoite rates was only detected in indoor resting An. coluzzii from the sahel savannah (5.0%) and forest (2.5%) zones.Conclusion: The study reports high outdoor resting densities of An. gambiae and An. coluzzii with high kdr west mutation frequencies, and persistence of malaria transmission indoors despite the use of LLINs and IRS. Continuous monitoring of changes in resting behavior of mosquitoes and implementation of complementary malaria control interventions are needed to target outdoor resting Anopheles mosquitoes in Ghana.

Resting Behavior of Blood-Fed Females and Host Feeding Preferences of Aedes aegypti (Diptera: Culicidae) Morphological Forms in Senegal

Aedes aegypti (Linnaeus) is the main vector of most arboviruses in tropical and subtropical urban areas. In West Africa, particularly in Senegal, domestic and wild populations have been described. Both Ae. aegypti aegypti (Aaa) and Ae. aegypti formosus (Aaf) were found in progenies of Ae. aegypti families from several localities of Senegal. However, nothing is known about their resting and trophic behavior, which are key data for vector control. To fill this gap, blood-fed mosquitoes were collected monthly indoors and outdoors with BackPack aspirators and BG-Sentinel 2 traps between July and November 2019 from four urban sites. The enzyme-linked immunosorbent assay technique was used to analyze blood-fed Aaa and Aaf specimens. Both forms were found resting in all investigated places with the highest proportions found in scrap metals (51.7% for Aaa and 44.1% for Aaf) and used tires (19.2% for Aaa and 26.1% for Aaf). Blood-fed Aaf females showed lower occupation of the indoors environment compared to Aaa. Overall, the percentages of single bloodmeals from human were 80.5% (916/1138) for Aaa and 71.1% (263/370) for Aaf. A low frequency of other domestic hosts, including bovine, ovine, and cat were detected for both forms. This study provides the first data on resting and trophic behavior of Aaa and Aaf in Senegal. Both forms showed differences in their resting behavior but fed primarily on human and highlight the risk of arboviruses transmission in urban areas.

High-Throughput Genotyping of Common Chromosomal Inversions in the Afrotropical Malaria Mosquito Anopheles Funestus

Polymorphic chromosomal inversions have been implicated in local adaptation. In anopheline mosquitoes, inversions also contribute to epidemiologically relevant phenotypes such as resting behavior. Progress in understanding these phenotypes and their mechanistic basis has been hindered because the only available method for inversion genotyping relies on traditional cytogenetic karyotyping, a rate-limiting and technically difficult approach that is possible only for the fraction of the adult female population at the correct gonotrophic stage. Here, we focus on an understudied malaria vector of major importance in sub-Saharan Africa, Anopheles funestus. We ascertain and validate tag single nucleotide polymorphisms (SNPs) using high throughput molecular assays that allow rapid inversion genotyping of the three most common An. funestus inversions at scale, overcoming the cytogenetic karyotyping barrier. These same inversions are the only available markers for distinguishing two An. funestus ecotypes that differ in indoor resting behavior, Folonzo and Kiribina. Our new inversion genotyping tools will facilitate studies of ecotypic differentiation in An. funestus and provide a means to improve our understanding of the roles of Folonzo and Kiribina in malaria transmission.

Interacting Effects of Polystyrene Microplastics and the Antidepressant Amitriptyline on Early Life Stages of Brown Trout (Salmo trutta f. fario)

Whether microplastics themselves or their interactions with chemicals influence the health and development of aquatic organisms has become a matter of scientific discussion. In aquatic environments, several groups of chemicals are abundant in parallel to microplastics. The tricyclic antidepressant amitriptyline is frequently prescribed, and residues of it are regularly found in surface waters. In the present study, the influence of irregularly shaped polystyrene microplastics (<50 µm), amitriptyline, and their mixture on early life-stages of brown trout were investigated. In a first experiment, the impacts of 100, 104, and 105 particles/L were studied from the fertilization of eggs until one month after yolk-sac consumption. In a second experiment, eggs were exposed in eyed ova stages to 105, 106 particles/L, to amitriptyline (pulse-spiked, average 48 ± 33 µg/L) or to two mixtures for two months. Microplastics alone did neither influence the development of fish nor the oxidative stress level or the acetylcholinesterase activity. Solely, a slight effect on the resting behavior of fry exposed to 106 particles/L was observed. Amitriptyline exposure exerted a significant effect on development, caused elevated acetylcholinesterase activity and inhibition of two carboxylesterases. Most obvious was the severely altered swimming and resting behavior. However, effects of amitriptyline were not modulated by microplastics.

Insecticide resistance in indoor and outdoor-resting Anopheles gambiae s.l. in Northern Ghana

Background: Selection pressure from continued exposure to insecticides drives the development of insecticide resistance and changes in resting behavior of malaria vectors, which may support residual transmission in several endemic settings. There is a need to understand how resistance drives changes in resting behavior within vector species. Here, we examined the association between insecticide resistance and resting behavior of Anopheles gambiae s.l. in Northern Ghana. Methods: Adult mosquitoes were collected both indoors and outdoors from two communities using mouth aspirators and pit shelters. F1 progenies from a subset of mosquitoes were exposed to dichloro diphenyl trichloroethane (DDT), deltamethrin, malathion and bendiocarb using WHO insecticide susceptibility tests. Insecticide resistance markers including voltage-gated sodium channel (Vgsc)- 1014F, Vgsc-1014S, Vgsc-1575Y, glutathione-S-transferase epsilon 2 (GSTe2)-114T and acetylcholinesterase (Ace1)-119S, as well as blood meal sources were investigated using PCR methods. Activities of metabolic enzymes, acetylcholine esterase (AChE), non-specific β-esterases, glutathione-S-transferase (GST) and monooxygenases (oxidases) were measured from unexposed F1 progenies using microplate assays. Results: Susceptibility of An. coluzzii to deltamethrin 24hr post-exposure was significantly higher in indoor (mortality=5%) than the outdoor (mortality=2.5%) populations (P=0.02). The mosquitoes were fully susceptible to malathion (mortality: indoor=98%, outdoor=100%). Susceptibility to DDT was significantly higher in outdoor (mortality=9%) than indoor (mortality=0%) mosquitoes (P=0.006). Mosquitoes were also found with suspected resistance to bendiocarb but mortality was not statistically different (mortality: indoor=90%, outdoor=95%. P=0.30). The frequencies of all resistance alleles were higher in F1 outdoor (0.11-0.85) than indoor (0.04-0.65) mosquito populations, while Vgsc-1014F in F0 An. gambiae s.s significantly associated with outdoor-resting behavior (P=0.01). Activities of non-specific β-esterase enzymes were significantly higher in outdoor than indoor mosquitoes (Mean enzyme activity: Outdoor=: 1.70/mg protein; Indoor=1.35/mg protein. P<0.0001). AChE activity was also more elevated in outdoor (0.62/mg protein) than indoor (0.57/mg protein) mosquitoes but this was not significant (P=0.08). Human blood index (HBI) was predominantly detected in indoor (18%) than the outdoor mosquito population (3%). Conclusions: These findings revealed higher phenotypic resistance in indoor than outdoor-resting mosquitoes, but genotypic and metabolic resistance levels were higher in outdoor than the indoor mosquito populations. However, the overall results did not establish that there was a significant preference of resistant malaria vectors to solely rest indoors or outdoors, but varied depending on the resistant alleles present. Indication that human and animal blood meal indices were more prevalent in indoor-resting mosquitoes was also shown. Continued monitoring of changes in resting behavior within An. gambiae s.l. populations is recommended.

Classification of Inter-Urban Highway Drivers’ Resting Behavior for Advanced Driver-Assistance System Technologies using Vehicle Trajectory Data from Car Navigation Systems

Fatigue-related crashes, which are mainly caused by drowsy or distracted driving, account for a significant portion of fatal accidents on highways. Smart vehicle technologies can address this issue of road safety to improve the sustainability of transportation systems. Advanced driver-assistance system (ADAS) can aid drowsy drivers by recommending and guiding them to rest locations. Past research shows a significant correlation between driving distance and driver fatigue, which has been actively studied in the analysis of resting behavior. Previous research efforts have mainly relied on survey methods at specific locations, such as rest areas or toll booths. However, such traditional methods, like field surveys, are expensive and often produce biased results, based on sample location and time. This research develops methods to better estimate travel resting behavior by utilizing a large-scale dataset obtained from car navigation systems, which contain 591,103 vehicle trajectories collected over a period of four months in 2014. We propose an algorithm to statistically categorize drivers according to driving distances and their number of rests. The main algorithm combines a statistical hypothesis test and a random sampling method based on the renowned Monte-Carlo simulation technique. We were able to verify that cumulative travel distance shares a significant relationship with one’s resting decisions. Furthermore, this research identifies the resting behavior pattern of drivers based upon their travel distances. Our methodology can be used by sustainable traffic safety operators to their driver guiding strategies criterion using their own data. Not only will our methodology be able to aid sustainable traffic safety operators in constructing their driver guidance strategies criterion using their own data, but it could also be implemented in actual car navigation systems as a mid-term solution. We expect that ADAS combined with the proposed algorithm will contribute to improving traffic safety and to assisting the sustainability of road systems.

A new genus and species of slug caterpillar (Lepidoptera: Limacodidae) from Taiwan

The genus Epsteinius Lin, Braby & Hsu gen. nov. is established to accommodate a new, diminutive species of limacodid moth, E. translucidus Lin, sp. nov., from central Taiwan. Although the new species is the sister taxon to Microleon Butler, 1885 according to molecular phylogenetic data, it differs fundamentally from that genus in adult morphology, wing color pattern, male and female genitalia, and larval morphology. Larvae of E. translucidus Lin, sp. nov. represent an interesting case in which crypsis is based on transparency through its body color, resting behavior, and feeding pattern.

Identifying spinner dolphin Stenella longirostris longirostris movement and behavioral patterns to inform conservation strategies in Maui Nui, Hawai‘i

Spinner dolphins Stenella longirostris longirostris in Hawai‘i exhibit a predictable diurnal behavioral pattern which makes them vulnerable to human disturbance: feeding at night in offshore waters and resting during daytime in bays. There is concern that human activity, such as swimming with and viewing wild dolphins from vessels, is altering their natural behavioral patterns and preventing them from having adequate rest. In light of this, state and federal management agencies are proposing enhanced protection measures. Research on spinner dolphins has largely focused off Hawai‘i Island and there are insufficient data from Maui Nui (Maui, Lāna‘i, Kaho‘olawe, and Moloka‘i) to inform appropriate management measures for the genetically distinct stock that resides around these islands. Using location data from 316 encounters between 1996 and 2019, we identified 2 hotspots for spinner dolphins within the region, located along west Maui and south Lāna‘i. The predominant behavior observed was traveling, and there was little resting behavior documented throughout the study period, with no resting behavior observed along the coastline of Maui. Our findings revealed that spinner dolphins use a wide variety of available habitat in Maui Nui and were observed resting both near the shore of Lāna‘i and in the channels between islands. Based upon these findings, the proposed lone area for closure in south Maui is inadequate for providing protection to spinner dolphins during resting hours, and we propose that the identified hotspots be considered as additional sites for closures, in addition to an approach limit for vessels transiting Maui Nui.