Addressing emerging issues in entomology: 2023 student debates.

The Entomological Society of America (ESA) Student Debates is an annual student competition at the ESA Annual Meeting organized by Student Debates Subcommittee (SDS) members of the ESA Student Affairs Committee. In conjunction with the 2023 ESA Annual Meeting theme, 'Insects and influence: Advancing entomology's impact on people and policy', the theme of this year's student debate was 'Addressing emerging issues in entomology'. With the aid of ESA membership, the SDS selected the following debate topics: (1) Should disclosure of artificial intelligence large language models in scientific writing always be required? and (2) Is it more important to prioritize honey bee or native pollinator health for long-term food security within North America? Four student teams from across the nation, composed of 3-5 student members and a professional advisor, were assigned a topic and stance. Over the course of 5 months, all team members researched and prepared for their assigned topic before debating live with an opposing team at the 2023 ESA Annual Meeting in National Harbor, Maryland. SDS members additionally prepared and presented introductions for each debate topic to provide unbiased backgrounds to the judges and audience for context in assessing teams' arguments. The result was an engaging discussion between our teams, judges, and audience members on emerging issues facing entomology and its impact on people and policy, such as scientific communication and food security, that brought attention to the complexities involved when debating topics concerning insects and influence.

Development of underground detection system using a metal detector and aluminum tag for, Copris ochus (Coleoptera: Scarabaeidae).

Tracking of soil-dwelling insects poses greater challenges compared to aboveground-dwelling animals in terrestrial systems. A metal detector system consisting of a commercially available detector and aluminum tags was developed for detecting dung beetle, Copris ochus Motschulsky (Coleoptera: Scarabaeidae). First, detection efficacy of the system was evaluated by varying volumes of aluminum tags attached on a plastic model of the insect and also by varying angles. Then, detection efficacy was evaluated by varying depths of aluminum-tagged models under soil in 2 vegetation types. Finally, the effects of tag attachment on C. ochus adults were assessed for survivorship, burrowing depth, and horizontal movement. Generally, an increase in tag volume resulted in greater detection distance in semi-field conditions. Maximum detection distance of aluminum tag increased up to 17 cm below soil surface as the tag size (0.5 × 1.0 cm [width × length]) and thickness (16 layers) were maximized, resulting in a tag weight of 31.4 mg, comprising ca. 9% of average weight of C. ochus adult. Furthermore, the detection efficacy did not vary among angles except for 90°. In the field, metal detectors successfully detected 5 aluminum-tagged models in 20 × 10 m (W × L) arena within 10 min with detection rates ≥85% for up to depth of 10 cm and 45%-60% at depth of 20 cm. Finally, aluminum tagging did not significantly affect survivorship and behaviors of C. ochus. Our study indicates the potential of metal detector system for tracking C. ochus under soil.

Initial laboratory validation of temperature development models for Necrodes littoralis L. (Staphylinidae: Silphinae).

Development models of necrophagous insects are applied in forensic entomology for post-mortem interval estimation. Such estimates may be used as scientific evidence in legal investigations. For this reason, it is important that the models are valid and that the expert witness is aware of their limitations. Necrodes littoralis L. (Staphylinidae: Silphinae) is a necrophagous beetle species that frequently colonizes human cadavers. Temperature models of development for the Central European population of these beetles were recently published. In this article, we present results of the laboratory validation study for these models. Errors of beetle age estimation differed significantly between the models. Thermal summation models yielded the most accurate estimates, and the isomegalen diagram least accurate estimates. Errors of the beetle age estimation varied across beetle developmental stages and rearing temperatures. In general, most development models of N. littoralis were satisfactorily accurate in estimating beetle age under laboratory conditions; therefore, the study provides initial evidence to support their validity in forensic cases.

Color preference of Sergentomyia minuta (Diptera: Phlebotominae) determined using Flebocollect Do It Yourself light traps based on LED technology.

Whether phlebotomine sand flies show a preference for different light colors remains controversial. As light-capture methods are widely used to study sand flies, knowing the visual stimuli they respond to could help the design of novel control tools to prevent their attraction to hosts. We have detected a significant preference of male Sergentomyia minuta for green and red light sources. Accordingly, male S. minuta were 2.16 and 2.01 times more likely to be lured by Flebocollect model traps with green and red diode-lights, respectively, than the commercial CDC traps. Flebocollect traps are homemade light traps developed through citizen science. Dipterans are widely considered unable to distinguish the color red so this finding was unexpected. To our knowledge, this is the first description of a color preference in a species of the genus Sergentomyia. Our research also confirms the great potential of Flebocollect light traps for use in medical entomology studies.

Which trap is best? Alternatives to outdoor human landing catches for malaria vector surveillance: a meta-analysis.

BACKGROUND: Human landing catches (HLC) are an entomological collection technique in which humans are used as attractants to capture medically relevant host-seeking mosquitoes. The use of this method has been a topic of extensive debate for decades mainly due to ethical concerns. Many alternatives to HLC have been proposed; however, no quantitative review and meta-analysis comparing HLC to outdoor alternative trapping methods has been conducted. METHODS: A total of 58 comparisons across 12 countries were identified. We conducted a meta-analysis comparing the standardized mean difference of Anopheles captured by HLC and alternative traps. To explain heterogeneity, three moderators were chosen for analysis: trap type, location of study, and species captured. A meta-regression was fit to understand how the linear combination of moderators helped in explaining heterogeneity. The possibility of biased results due to publication bias was also explored. RESULTS: Random-effects meta-analysis showed no statistically significant difference in the mean difference of Anopheles collected. Moderator analysis was conducted to determine the effects of trap type, geographical location of study, and the species of Anopheles captured. On average, tent-based traps captured significantly more Anopheles than outdoor HLC (95% CI: [- .9065, - 0.0544]), alternative traps in Africa captured on average more mosquitoes than outdoor HLC (95% CI: [- 2.8750, - 0.0294]), and alternative traps overall captured significantly more Anopheles gambiae s.l. than outdoor HLC (95% CI: [- 4.4613, - 0.2473]) on average. Meta-regression showed that up to 55.77% of the total heterogeneity found can be explained by a linear combination of the three moderators and the interaction between trap type and species. Subset analysis on An. gambiae s.l. showed that light traps specifically captured on average more of this species than HLC (95% CI: [- 18.3751, - 1.0629]). Publication bias likely exists. With 59.65% of studies reporting p-values less than 0.025, we believe there is an over representation in the literature of results indicating that alternative traps are superior to outdoor HLC. CONCLUSIONS: Currently, there is no consensus on a single "magic bullet" alternative to outdoor HLC. The diversity of many alternative trap comparisons restricts potential metrics for comparisons to outdoor HLC. Further standardization and specific question-driven trap evaluations that consider target vector species and the vector control landscape are needed to allow for robust meta-analyses with less heterogeneity and to develop data-driven decision-making tools for malaria vector surveillance and control.

Preference Provides a Plethora of Problems (Don't Panic).

This review was solicited as an autobiography. The "problems" in my title have two meanings. First, they were professional difficulties caused by my decision to study oviposition preferences of butterflies that were not susceptible to traditional preference-testing designs. Until I provided video, my claim that the butterflies duplicate natural post-alighting host-assessment behavior when placed on hosts by hand was not credible, and the preference-testing technique that I had developed elicited skepticism, anger, and derision. The second meaning of "problems" is scientific. Insect preference comes with complex dimensionality that interacts with host acceptability. Part Two of this review describes how my group's work in this area has revealed unexpected axes of variation in plant-insect interactions-axes capable of frustrating attempts to derive unequivocal conclusions from apparently sensible experimental designs. The possibility that these complexities are lurking should be kept in mind as preference and performance experiments are devised.

Reduced exposure to malaria vectors following indoor residual spraying of pirimiphos-methyl in a high-burden district of rural Mozambique with high ownership of long-lasting insecticidal nets: entomological surveillance results from a cluster-randomized trial.

BACKGROUND: The need to develop new products and novel approaches for malaria vector control is recognized as a global health priority. One approach to meeting this need has been the development of new products for indoor residual spraying (IRS) with novel active ingredients for public health. While initial results showing the impact of several of these next-generation IRS products have been encouraging, questions remain about how to best deploy them for maximum impact. To help address these questions, a 2-year cluster-randomized controlled trial to measure the impact of IRS with a microencapsulated formulation of pirimiphos-methyl (PM) in an area with high ownership of long-lasting insecticidal nets (LLINs) was conducted in a high-transmission district of central Mozambique with pyrethroid resistant vectors. Presented here are the results of the vector surveillance component of the trial. METHODS: The 2 year, two-armed trial was conducted in Mopeia District, Zambezia Province, Mozambique. In ten sentinel villages, five that received IRS with PM in October-November 2016 and again in October-November 2017 and five that received no IRS, indoor light trap collections and paired indoor-outdoor human landing collections catches (HLCs) were conducted monthly from September 2016 through October 2018. A universal coverage campaign in June 2017, just prior to the second spray round, distributed 131,540 standard alpha-cypermethrin LLINs across all study villages and increased overall net usage rates in children under 5 years old to over 90%. RESULTS: The primary malaria vector during the trial was Anopheles funestus sensu lato (s.l.), and standard World Health Organization (WHO) tube tests with this population indicated variable but increasing resistance to pyrethroids (including alpha-cypermethrin, from > 85% mortality in 2017 to 7% mortality in 2018) and uniform susceptibility to PM (100% mortality in both years). Over the entire duration of the study, IRS reduced An. funestus s.l. densities by 48% (CI95 33-59%; p < 0.001) in indoor light traps and by 74% (CI95 38-90%; p = 0.010) during indoor and outdoor HLC, though in each study year reductions in vector density were consistently greatest in those months immediately following the IRS campaigns and waned over time. Overall there was no strong preference for An. funestus to feed indoors or outdoors, and these biting behaviours did not differ significantly across study arms: observed indoor-outdoor biting ratios were 1.10 (CI95 1.00-1.21) in no-IRS villages and 0.88 (CI95 0.67-1.15) in IRS villages. The impact of IRS was consistent in reducing HLC exposures both indoors (75% reduction: CI95 47-88%; p = 0. < 0.001) and outdoors (68% reduction: CI95 22-87%; p = 0.012). While substantially fewer Anopheles gambiae s.l. were collected during the study, trends show a similar impact of IRS on this key vector group as well, with a 33% (CI95 7-53%; p = 0.019) reduction in mosquitoes collected in light traps and a non-statistically significant 39% reduction (p = 0.249) in HLC landing rates. CONCLUSION: IRS with PM used in addition to pyrethroid-only LLINs substantially reduced human exposures to malaria vectors during both years of the cluster-randomized controlled trial in Mopeia-a high-burden district where the primary vector, An. funestus s.l., was equally likely to feed indoors or outdoors and demonstrated increasing resistance to pyrethroids. Findings suggest that IRS with PM can provide effective vector control, including in some settings where pyrethroid-only ITNs are widely used. Trial registration clinicaltrials.gov , NCT02910934. Registered 22 September 2016, https://www.clinicaltrials.gov/ct2/show/NCT02910934.

Simple method to detect and to isolate entomopathogenic fungi (Hypocreales) from mosquito larvae.

Entomopathogenic fungi are important agents for mosquito vector control. We report on the utility of a simple method to detect fungi on living larvae of Aedes aegypti that had been exposed to a fungal entomopathogen. Four species of the hypocrealean genera Metarhizium, Beauveria, Tolypocladium and Culicinomyces, known for their larvicidal activity against mosquito species, were tested. Living larvae previously exposed to a suspension of different conidial concentrations were set directly into the surface water film on non-nutritive agar supplemented with chloramphenicol, thiabendazole and crystal violet and then incubated. Except for C. clavisporus ARSEF 964 (which developed and produced conidia mostly inside the cadaver rather than on its surface in the present study), this method favored external fungal development and conidiogenesis on larvae of different instars after death. The dead larva on the water agar represents the unique and specific source of nutrition for the fungus that killed it. The technique facilitates the detection and posterior isolation of entomopathogenic fungi, and offers a compact, convenient, and rapid means to survey larval mosquito populations for fungal pathogens at the field.

The Use of Frozen, Food-Grade Blood to Successfully Maintain Colonies of Four Species of Mosquitoes (Diptera: Culicidae).

An essential component of all mosquito-rearing activities is the act of blood-feeding the mosquitoes (Diptera: Culicidae). Many options exist for this purpose including live host animals and a diverse array of artificial-feeding methods. Most of the published artificial-feeding methods involve expensive materials, custom-built devices, or are labor-intensive. All of the previously published methods utilize blood sources, which are either expensive, or difficult to obtain. Additionally, much of the research into artificial blood-feeding methods for mosquitoes has focused on two species: Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse). This article presents a modified artificial blood-feeding method that uses affordable and easily sourced materials, does not require any technical knowledge to assemble, and requires minimal time and effort. The combination of inexpensive aluminum plates, Parafilm and polytetrafluoroethylene tape membranes, an electric germination mat, and frozen, food-grade blood produces exceptional feeding rates and abundant egg production. The method has been used for 2 yr at the Lee County Mosquito Control District to successfully maintain laboratory colonies of four species of mosquito: Ae. aegypti, Ae. albopictus, Aedes taeniorhynchus (Wiedemann), and Culex quinquefasciatus (Say). Variations of this method are reported, which can be used for wild and laboratory colonies of multiple species. This modified method is highly accessible for any small-scale mosquito rearing facility with labor or budgetary constraints.

A Visual Tracking System for Honey Bee (Hymenoptera: Apidae) 3D Flight Trajectory Reconstruction and Analysis.

We describe the development, field testing, and results from an automated 3D insect flight detection and tracking system for honey bees (Apis mellifera L.) (Hymenoptera: Apidae) that is capable of providing remarkable insights into airborne behavior. It comprises two orthogonally mounted video cameras with an observing volume of over 200 m3 and an offline analysis software system that outputs 3D space trajectories and inflight statistics of the target honey bees. The imaging devices require no human intervention once set up and are waterproof, providing high resolution and framerate videos. The software module uses several forms of modern image processing techniques with GPU-enabled acceleration to remove both stationary and moving artifact while preserving flight track information. The analysis system has thus far provided information not only on flight statistics (such as speeds and accelerations), but also on subtleties associated with flight behavior by generating heat maps of density and classifying flight patterns according to patrol and foraging behavior. Although the results presented here focus on behavior in the locale of a beehive, the system could be adapted to study a wide range of airborne insect activity.

Testing the Accuracy of Vegetation-Based Ecoregions for Predicting the Species Composition of Blow Flies (Diptera: Calliphoridae).

To properly define ecoregions, specific criteria such as geology, climate, or species composition (e.g., the presence of endemic species) must be taken into account to understand distribution patterns and resolve ecological biogeography questions. Since the studies on insects in Baja California are scarce, and no fine-scale ecoregions based on the region's entomofauna is available, this study was designed to test whether the ecoregions based on vegetation can be used for insects, such as Calliphoridae. Nine collecting sites distributed along five ecoregions were selected, between latitudes 29.6° and 32.0°N. In each site, three baited traps were used to collect blow flies from August 2017 to June 2019 during summer, winter, and spring. A total of 30,307 individuals of blow flies distributed in six genera and 13 species were collected. The most abundant species were Cochliomyia macellaria (Fabricius), Phormia regina (Meigen), and Chrysomya rufifacies (Macquart). The composition of the Calliphoridae community was different between the localities and three general groups have been distinguished, based on the species composition similarity (ANOSIM) results: Gulf-Desert, Mountains, and Pacific-Center. The vegetation-based ecoregions only reflect the blow fly species' distributions to a certain extent, meaning that care must be taken when undertaking ecological biogeographical studies using regionalization based on organisms other than the focal taxa because vegetation does not always reflect fauna species composition.

Impact of Agro-industrial Byproducts on Bioconversion, Chemical Composition, in vitro Digestibility, and Microbiota of the Black Soldier Fly (Diptera: Stratiomyidae) Larvae.

The interest in using byproducts from agro-food industries as a rearing substrate for insects is increasing rapidly. We investigated the influence of byproducts of vegetal origin (okara-a byproduct of soy milk production, maize distillers with solubles, brewer's grains), used as rearing diet for black soldier fly larvae (BSFL), on the following parameters: biomass production, substrate reduction (SR), nutritional profile and in vitro digestibility, and larval gut microbiota. Hen diet was used as a control substrate. The highest larval biomass was collected on maize distillers, whereas the highest SR was observed on okara. The rearing substrate affected ash, ether extract, and chitin larval content. The BSFL reared on okara were characterized by a lower lauric acid content (17.6% of total fatty acids). Diets also influenced in vitro crude protein digestibility (%) for monogastrics, with the highest values for BSFL reared on maize distillers (87.8), intermediate for brewer's grains and okara BSFL, and the lowest for hen BSFL (82.7). The nutritive value for ruminants showed a lower Net Energy for lactation for BSFL reared on hen diet than okara and dried maize distillers BSFL. The different byproducts showed an influence on the larval gut microbiota, with a major bacterial complexity observed on larvae fed with the hen diet. The neutral detergent fiber concentration of dietary substrate was negatively correlated with Firmicutes and Actinobacteria relative abundance. Insects valorized byproducts converting them into high-value larval biomass to be used for feed production. The results evidenced the effects of the tested byproducts on the measured parameters, underling the chemical composition importance on the final insect meal quality.

A Review of Insect Monitoring Approaches with Special Reference to Radar Techniques.

Drastic declines in insect populations are a vital concern worldwide. Despite widespread insect monitoring, the significant gaps in the literature must be addressed. Future monitoring techniques must be systematic and global. Advanced technologies and computer solutions are needed. We provide here a review of relevant works to show the high potential for solving the aforementioned problems. Major historical and modern methods of insect monitoring are considered. All major radar solutions are carefully reviewed. Insect monitoring with radar is a well established technique, but it is still a fast-growing topic. The paper provides an updated classification of insect radar sets. Three main groups of insect radar solutions are distinguished: scanning, vertical-looking, and harmonic. Pulsed radar sets are utilized for all three groups, while frequency-modulated continuous-wave (FMCW) systems are applied only for vertical-looking and harmonic insect radar solutions. This work proves the high potential of radar entomology based on the growing research interest, along with the emerging novel setups, compact devices, and data processing approaches. The review exposes promising insect monitoring solutions using compact radar instruments. The proposed compact and resource-effective setups can be very beneficial for systematic insect monitoring.

Saving time maintaining reliability: a new method for quantification of Tetranychus urticae damage in Arabidopsis whole rosettes.

BACKGROUND: The model species Tetranychus urticae produces important plant injury and economic losses in the field. The current accepted method for the quantification of the spider mite damage in Arabidopsis whole rosettes is time consuming and entails a bottleneck for large-scale studies such as mutant screening or quantitative genetic analyses. Here, we describe an improved version of the existing method by designing an automatic protocol. The accuracy, precision, reproducibility and concordance of the new enhanced approach are validated in two Arabidopsis accessions with opposite damage phenotypes. Results are compared to the currently available manual method. RESULTS: Image acquisition experiments revealed that the automatic settings plus 10 values of brightness and the black background are the optimal conditions for a specific recognition of spider mite damage by software programs. Among the different tested methods, the Ilastik-Fiji tandem based on machine learning was the best procedure able to quantify the damage maintaining the differential range of damage between accessions. In addition, the Ilastik-Fiji tandem method showed the lowest variability within a set of conditions and the highest stability under different lighting or background surroundings. Bland-Altman concordance results pointed out a negative value for Ilastik-Fiji, which implies a minor estimation of the damage when compared to the manual standard method. CONCLUSIONS: The novel approach using Ilastik and Fiji programs entails a great improvement for the quantification of the specific spider mite damage in Arabidopsis whole rosettes. The automation of the proposed method based on interactive machine learning eliminates the subjectivity and inter-rater-variability of the previous manual protocol. Besides, this method offers a robust tool for time saving and to avoid the damage overestimation observed with other methods.

Evaluation of human-baited double net trap and human-odour-baited CDC light trap for outdoor host-seeking malaria vector surveillance in Kenya and Ethiopia.

BACKGROUND: Surveillance of outdoor host-seeking malaria vectors is crucial to monitor changes in vector biting behaviour and evaluate the impact of vector control interventions. Human landing catch (HLC) has been considered the most reliable and gold standard surveillance method to estimate human-biting rates. However, it is labour-intensive, and its use is facing an increasing ethical concern due to potential risk of exposure to infectious mosquito bites. Thus, alternative methods are required. This study was conducted to evaluate the performance of human-odour-baited CDC light trap (HBLT) and human-baited double net trap (HDNT) for outdoor host-seeking malaria vector surveillance in Kenya and Ethiopia. METHODS: The sampling efficiency of HBLT and HDNT was compared with CDC light trap and HLC using Latin Square Design in Ahero and Iguhu sites, western Kenya and Bulbul site, southwestern Ethiopia between November 2015 and December 2018. The differences in Anopheles mosquito density among the trapping methods were compared using generalized linear model. RESULTS: Overall, 16,963 female Anopheles mosquitoes comprising Anopheles gambiae sensu lato (s.l.), Anopheles funestus s.l., Anopheles pharoensis, Anopheles coustani and Anopheles squamosus were collected. PCR results (n = 552) showed that Anopheles arabiensis was the only member of An. gambiae s.l. in Ahero and Bulbul, while 15.7% An. arabiensis and 84.3% An. gambiae sensu stricto (s.s.) constituted An. gambiae s.l. in Iguhu. In Ahero, HBLT captured 2.23 times as many An. arabiensis and 2.11 times as many An. funestus as CDC light trap. In the same site, HDNT yielded 3.43 times more An. arabiensis and 3.24 times more An. funestus than HBLT. In Iguhu, the density of Anopheles mosquitoes did not vary between the traps (p > 0.05). In Bulbul, HBLT caught 2.19 times as many An. arabiensis as CDC light trap, while HDNT caught 6.53 times as many An. arabiensis as CDC light trap. The mean density of An. arabiensis did not vary between HDNT and HLC (p = 0.098), whereas the HLC yielded significantly higher density of An. arabiensis compared to HBLT and CDC light trap. There was a significant density-independent positive correlation between HDNT and HLC (r = 0.69). CONCLUSION: This study revealed that both HBLT and HDNT caught higher density of malaria vectors than conventional CDC light trap. Moreover, HDNT yielded a similar vector density as HLC, suggesting that it could be an alternative tool to HLC for outdoor host-seeking malaria vector surveillance.

Endosymbiotic Bacteria Are Prevalent and Diverse in Agricultural Spiders.

Maternally inherited bacterial endosymbionts are common in arthropods, but their distribution and prevalence are poorly characterized in many host taxa. Initial surveys have suggested that vertically transmitted symbionts may be particularly common in spiders (Araneae). Here, we used diagnostic PCR and high-throughput sequencing to evaluate symbiont infection in 267 individual spiders representing 14 species (3 families) of agricultural spiders. We found 27 operational taxonomic units (OTUs) that are likely endosymbiotic, including multiple strains of Wolbachia, Rickettsia, and Cardinium, which are all vertically transmitted and frequently associated with reproductive manipulation of arthropod hosts. Additional strains included Rickettsiella, Spiroplasma, Rhabdochlamydia, and a novel Rickettsiales, all of which could range from pathogenic to mutualistic in their effects upon their hosts. Seventy percent of spider species had individuals that tested positive for one or more endosymbiotic OTUs, and specimens frequently contained multiple symbiotic strain types. The most symbiont-rich species, Idionella rugosa, had eight endosymbiotic OTUs, with as many as five present in the same specimen. Individual specimens within infected spider species had a variety of symbiotypes, differing from one another in the presence or absence of symbiotic strains. Our sample included both starved and unstarved specimens, and dominant bacterial OTUs were consistent per host species, regardless of feeding status. We conclude that spiders contain a remarkably diverse symbiotic microbiota. Spiders would be an informative group for investigating endosymbiont population dynamics in time and space, and unstarved specimens collected for other purposes (e.g., food web studies) could be used, with caution, for such investigations.

Anopheles gambiae s.l (Diptera: Culicidae) seasonal abundance, abdominal status and parity rates in Metema-Armachiho lowland, Northwest Ethiopia.

BACKGROUND: Malaria is a life threating vector borne disease caused by different Plasmodium parasites. Metema and Armachiho are two of the top five malaria endemic areas among the districts of Amhara region in Ethiopia. Transmission pattern is seasonal and migrant laborers who visit these areas for employment in mechanized agriculture are highly affected. The aim of this study was to investigate seasonal abundance, abdominal status and parity rate of An.gambiae s.l in Metema-Armachiho lowlands, Northwest Ethiopia. METHOD: A 1 year longitudinal entomological study was conducted in Metema-Armachiho lowlands from June 2016-May 2017. Mosquitoes were collected using CDC-light traps in indoor and outdoor sites for four consecutive days in each month. A total of eight standard battery operated CDC-light traps were used to collect mosquitoes. Female mosquitoes were classed as unfed, fed or gravid under a dissecting microscope. The ovaries of all unfed An.gambiae s.l mosquitoes were examined for evidence of parity. Data were entered and analyzed using SPSS-20 software. Chi-square test was applied to show significant difference between variables. P-value< 0.05 was taken as statistically significant. RESULTS: Of the total 1253 mosquitoes collected, 713 (552 female, 161 male) were culex and 540 (501 female, 39 male) were An.gambiae s.l. About 50.9% were collected in June-August 2016, 21.7% in September-November 2016, 12.0% in December 2016-February 2017 and 15.4% in March to May 2017. Of the total, 57.2 and 42.8% of the An.gambiae s.l were collected from indoor and outdoor sites respectively. Of the total females collected, 76.8% were unfed; of which 69.4% were parous. Significantly higher number of female An.gambiae s.l were collected in indoor and there was significant difference in abdominal status of An.gambiae s.l mosquitoes collected in different season (P < 0.05). CONCLUSIONS: Highest number of An.gambiae s.l was observed from June-August followed by September-November. The parity rate of An.gambiae s.l was high and there was significant difference in abdominal status of An.gambiae s.l collected in different season.

Identification of mixed and successive blood meals of mosquitoes using MALDI-TOF MS protein profiling.

BACKGROUND: The accurate and rapid identification of mosquito blood meals is critical to study the interactions between vectors and vertebrate hosts and, subsequently, to develop vector control strategies. Recently, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiling has been shown to be a reliable and effective tool for identifying single blood meals from mosquitoes. METHODS: In this study, we developed MALDI-TOF MS profiling protocols to identify Anopheles gambiae Giles, Anopheles coluzzii and Aedes albopictus mosquitoes' mixed blood meals and the last of successive blood meals. The mosquitoes were either successively artificially fed with distinct host bloods or engorged with mixed bloods from distinct vertebrate hosts, such as humans, sheep and dogs. RESULTS: Blind test analyses revealed a correct identification of mixed blood meals from mosquitoes using MALDI-TOF MS profiling. The 353 MS spectra from mixed blood meals were identified using log score values >1.8. All MS spectra (n = 244) obtained from mosquitoes' successive blood meals were reproducible and specific to the last blood meal, suggesting that the previous blood meals do not have an impact on the identification of the last one. CONCLUSION: MALDI-TOF MS profiling approach appears to be an effective and robust technique to identify the last and mixed blood meals during medical entomological surveys.

Estimating tsetse fertility: daily averaging versus periodic larviposition.

When computing mean daily fertility in adult female tsetse, the common practice of taking the reciprocal of the interlarval period (called averaged fertility) was compared with the method of taking the sum of the products of daily fertility and adult survivorship divided by the sum of daily survivorships (called periodic fertility). The latter method yielded a consistently higher measure of fertility (approximately 10% for tsetse) than the former method. A conversion factor was calculated to convert averaged fertility to periodic fertility. A feasibility criterion was determined for the viability of a tsetse population. Fertility and survivorship data from tsetse populations on Antelope Is. and Redcliff Is., both in Zimbabwe, were used to illustrate the feasibility criterion, as well as the limitations imposed by survivorship and fertility on the viability of tsetse populations. The 10% difference in fertility between the two methods of calculation makes the computation of population feasibility with some parameter combinations sometimes result in a wrong answer. It also underestimates both sterile male release rates required to eradicate a pest population, as well as the speed of resurgence if an eradication attempt fails.

A Guide and Toolbox to Replicability and Open Science in Entomology.

The ability to replicate scientific experiments is a cornerstone of the scientific method. Sharing ideas, workflows, data, and protocols facilitates testing the generalizability of results, increases the speed that science progresses, and enhances quality control of published work. Fields of science such as medicine, the social sciences, and the physical sciences have embraced practices designed to increase replicability. Granting agencies, for example, may require data management plans and journals may require data and code availability statements along with the deposition of data and code in publicly available repositories. While many tools commonly used in replicable workflows such as distributed version control systems (e.g., 'git') or script programming languages for data cleaning and analysis may have a steep learning curve, their adoption can increase individual efficiency and facilitate collaborations both within entomology and across disciplines. The open science movement is developing within the discipline of entomology, but practitioners of these concepts or those desiring to work more collaboratively across disciplines may be unsure where or how to embrace these initiatives. This article is meant to introduce some of the tools entomologists can incorporate into their workflows to increase the replicability and openness of their work. We describe these tools and others, recommend additional resources for learning more about these tools, and discuss the benefits to both individuals and the scientific community and potential drawbacks associated with implementing a replicable workflow.