Wing Length Research Articles

First report of Culex (Lophoceraomyia) cinctellus in Sri Lanka: Evidence from morphological and molecular analysis

Objective: To confirm the presence of Culex (Cx.) (Lophoceraomyia) cinctellus in Sri Lanka using morphological and molecular evidence. Methods: From October 2019 to April 2020, mosquito surveillance was conducted fort-nightly in the Banduragoda Public Health Inspector area. Larvae were collected using standard siphoning methods, while adults were sampled using Cattle Baited Trap, Gravid Traps, Light Traps, Bird-Baited Traps, Dog Baited Traps, and diurnal human landing collections. Specimens were transported to the Entomology Laboratory at the Medical Research Institute for identification. Morphological identification was performed using standard taxonomic keys. Molecular confirmation was achieved through DNA sequencing of mosquito head and thoracic regions, followed by sequence analysis using NCBI BLAST and Geneious software (version 7.1.3). Results: Adults of Cx. cinctellus were identified in Bird-Baited Traps and human bait collections. Unique morphological characteristics, including well-developed pulvilli, wing vein 1A ending before the apex of cross vein mcu, basal transverse pale bands on abdominal terga, and two labial basal setae on the proboscis, confirmed species identity. Morphometric measurements included mean thoracic length (0.58±0.02) mm, thoracic width (0.63±0.02) mm, abdominal length (2.15±0.03) mm, abdominal width (0.61±0.01) mm, and wing length (2.91±0.02) mm. Molecular analysis corroborated the morphological identification, affirming the species as Cx. cinctellus. COI sequences of the collected specimen (452 bp) were confirmed as Cx. cinctellus for sequence identity by BLAST and BOLD analysis. These sequences were subsequently deposited in GenBank under the accession number OR225623.1. Conclusions: This study documents the first occurrence of Cx. cinctellus in Sri Lanka, highlighting the need to enhance entomological surveillance to monitor its dispersal and population dynamics.

The effect of the egg storage time on the hatching success, morphometrics of larvae, and adult emergence of Aedes aegypti and Aedes albopictus

AbstractAedes aegypti (L.) and Aedes albopictus (Skuse) are important vectors that transmit arboviruses such as Dengue virus (DENV), Yellow fever virus (YFV), Chikungunya virus (CHIKV), and Zika virus (ZIKV). Focusing on the unique characteristics of their eggs to withstand dry conditions, this study aimed to elucidate the viability of dry‐stored eggs and the subsequent morphometric changes in larvae and adult mosquitoes upon rehydration. Experiments were carried out using ovitrap‐collected eggs, which were reared in the laboratory until adult emergence. The resulting mosquitoes were fed on cattle blood and sugar solution and allowed to lay eggs on the paper strips, which were air‐dried and stored for 6 months. The viability of these eggs was tested monthly. The head length and total length of fourth‐instar larvae and head, thorax, abdomen, wing length, wing width, and total length of adults were measured, including the days needed for adult emergence. The results imply that the prolonging of the egg storage period led to a decline in viability, delayed emergence of adults, and a negative impact on the morphometrics of fourth‐instar larvae in both Ae. aegypti and Ae. albopictus. Nonetheless, the morphometrics of the adults did not exhibit any significant differences compared to those of fresh eggs.

Shrinking body size under climate warming is not associated with selection for smaller individuals in a migratory bird.

How species are responding to climate change is a key topic in evolutionary ecology. Increasing temperatures are expected to affect phenotypic traits involved in thermoregulation, thus decreasing body size and/or increasing body appendages associated with heat exchange, as predicted by Bergmann's and Allen's rules. Results from long-term studies of variation in morphology over time have generally provided results supporting these predictions. However, two outstanding questions are frequently raised in studies relating changes in phenotypes to increasing temperatures: (1) whether such changes involve a shift in animal shape through the non-proportional variation of different body parts; and (2) whether they result from adaptive evolutionary responses. Relying on capture-recapture histories of almost 9000 breeding individuals from a declining Italian population of an Afro-Palearctic migratory bird, the barn swallow (Hirundo rustica), we documented a decrease in some body size traits (body mass, keel and wing length) over a 31-year period (1993-2023), with body mass declining the most (up to 4.0% in males). However, this was not the case for bill and partly tarsus length. Intra-individual lifelong changes in morphological traits of sexually mature birds showed only a limited contribution to trends over time in phenotypically plastic morphological traits. Viability and fecundity selection analyses revealed that smaller individuals did not enjoy greater success compared to larger ones. For some traits, the opposite was actually the case. The shifts in body size and, partly, shape over time we observed were coherent with predictions deriving from Bergmann's and Allen's rules. Yet, natural selection did not consistently favour smaller individuals. We thus call for caution in interpreting recent decreases in body size as adaptive evolutionary responses to climate warming, as they may rather reflect phenotypically plastic responses to changing climatic/environmental conditions occurring during early ontogenetic stages.

Investigation of The Morphological Characteristics of Cones and Seeds of The Scots pine (Pinus sylvestris L.) in Natural Populations in Northwestern Türkiye

Aim of study: The aim of this study is to investigate the morphological characteristics of Scots pine cones and seeds in a variety of natural populations. Area of study: The study was carried out in 12 different natural Scots pine populations in the Western Black Sea Region of Türkiye. Material and method: A total of 600 cone and 1727 seed samples were collected from 12 natural Scots pine populations. Multivariate Analysis of Variance and post-hoc test were used to investigate differences in cone and seed characteristics including cone length (cl), cone diameter (cd), cl/cd ratio, seed length (sl), seed width (sw), sl/sw ratio, seed wing length (wl), seed wing width (ww) and wl/ww between populations. Main results: Cone and seed morphological characteristics of Scots pine individuals showed statistically significant differences between populations. The Dedemdağ (Tosya) population had the largest cones in terms of both diameter and length, while the Kızılkaya (Yenice) and Kartalkaya (Aladağ) populations had the smallest cones. Research highlights: The protection of natural populations of Scots pine is very important, especially for the conservation of seed resources. This study provides important information about potential Scots pine populations in Northwestern Türkiye.

Heritability of morphological traits and mating probability of males derived from successful parents of the Anastrepha ludens Tapachula‐7 strain

AbstractHeritability measures the proportion of variation in a phenotypic trait within a population attributable to genetic differences among individuals. It also reflects genetic variation that can be passed from one generation to the next, and that can be used to predict the effectiveness of selection on traits involved in enhancing individual performance in genetic breeding programs. In fruit flies, females may select males based on chemical, acoustical, and visual signals as well as physical or morphological characteristics. However, the extent to which genetic variation underlying these traits is passed on to offspring is still largely unknown. Here, we used flies from a genetic sexing strain of Anastrepha ludens (Diptera: Tephritidae), known as Tapachula 7, to determine and compare the probability of mating for males derived from parents where mating interactions were successful vs. those that were not under field cage conditions. Offspring of both groups were propagated in the laboratory, and results showed that males derived from successful parents had a greater probability of mating compared to males derived from parents that were unsuccessful in the field cages. We also estimated the heritability of four morphological traits (pupal weight, thorax length, head width, and wing length) related to male mating success. The morphological characters of pupal weight and thorax length produced relatively low heritability values compared with medium and higher heritability values for head width and wing length, respectively. This study demonstrated that a degree of genetic association exists between reproductive success and some morphological traits associated with mating success in males of the Tapachula‐7 strain of A. ludens. The possibility of using these results for improving the sexual performance of sterile males through directed selection is discussed.

Effects of larval feeding amount on development and deltamethrin resistance in Aedes albopictus

To investigate how larval feeding regimens influence development and deltamethrin resistance of Aedes albopictus to provide evidence for standardizing larval feeding protocols in studies of insecticide resistance. Aedes albopictus larvae of a laboratory resistant strain were divided into 3 groups (n=500) and reared with high, medium, and low food availability (100, 50, or 25 mg daily for the 1st and 2nd instars, and 500 mg 250, or 125 mg daily for 3rd and 4th instars). The developmental time, pupation rate, adult emergence rate, adult body weight, and wing length were recorded in each group, and deltamethrin resistance of the mosquitoes was assessed using larval bioassays and contact tube tests for adults. Significant developmental differences were observed across the 3 feeding groups. Larval development time decreased as the food availability increased, and both high- and low-food groups showed reduced pupation rates (χ²=16.282, 7.440) and emergence rates (χ²=4.093, 6.977) compared to the medium-food group. Adult body weight and wing length were positively correlated with the amount of larval food intake (P<0.05). In high, medium and low food intake groups, larval LC50 values for deltamethrin were 0.110, 0.072 and 0.064 mg/L, adult KDT50 values were 97.404, 68.964 and 65.005 min, and adult mosquitoe mortality rates at 24 h after deltamethrin exposure were 12%, 16% and 19%, respectively. The feeding amount during larval stage significantly impacts the development and deltamethrin resistance of Aedes albopictus, suggesting the importance of standardization of larval nutrition for ensuring comparability of resistance test data across laboratories.

Like mother, like daughter? Phenotypic plasticity, environmental covariation, and heritability of size in a parthenogenetic wasp.

Parthenogenetic wasps provide an ideal natural experiment to study the heritability, plasticity, and microevolutionary dynamics of body size. Dinocampus coccinellae (Hymenoptera:Braconidae, Euphorinae) is a solitary, generalist braconid parasitoid wasp that reproduces through thelytokous parthenogenesis, and parasitizes over fifty diverse species of coccinellid ladybeetles worldwide as hosts. Here we designed an experiment with parthenogenetic lines of D. coccinellae presented with three different host ladybeetle species of varying sizes, across multiple generations to investigate heritability, and plasticity of body size measured via a combination of morphometric variables such as thorax width, abdominal width, and wing length in D. coccinellae. We expected positively correlated parent-offspring parasitoid regressions, indicative of heritable size variation, from unilineal (parent and offspring reared on same host species) lines, since these restrict environmental variation in phenotypes. In contrast, because multilineal (parent and offspring reared on different host species) lines would induce phenotypic plasticity of clones reared in varying environments, we expected negatively correlated parent-offspring parasitoid regressions. Our results indicate (1) little heritable variation in body size, (2) strong independence of offspring size on the host environment, (3) small mothers produce larger offspring, and vice versa, independent of host. We then model the evolution of size and host-shifting under a constrained fecundity advantage model of Cope's Law using a Hidden Markov Model, showing that D. coccinellae likely has fitness advantages to maintain plasticity in body size despite parthenogenetic reproduction.

Urban trace metal contamination is negatively associated with condition and wing morphology in a common waterbird.

Urban trace metal contamination is negatively associated with condition and wing morphology in a common waterbird.

Exploring stress and morphology in two songbird species across urban, agricultural, and natural habitats on San Cristobal Island, Galapagos

Land use changes can have morphological and physiological impacts on wildlife. This study aimed to explore the influence of anthropogenic land use on the morphology and corticosterone concentrations in two songbirds endemic to the Galapagos archipelago: the granivorous Small Ground Finch Geospiza fuliginosa and the insectivorous Galapagos Yellow Warbler Setophaga petechia aureola in San Cristobal Island. Birds were caught and measured between June and August 2018 and June and July 2019 across four areas with different human land uses: urban green areas in the coastal town of Puerto Baquerizo Moreno, natural deciduous forest in the lowlands, agricultural areas in the highlands, and seasonal evergreen forest in the highlands. Morphological comparisons among study areas were made using ANOVA or the Kurskall-Wallis test. Corticosterone levels obtained from tail feathers were measured with an ELISA test. Linear regression models were employed to explore the effects of the different human land uses on corticosterone concentrations. For G. fuliginosa, we found significant differences (p < 0.05) in weight, wing, and tarsus length between natural and disturbed habitats. The linear regression results showed higher corticosterone concentrations in urban G. fuliginosa than those in agricultural and natural habitats. Additionally, higher corticosterone concentrations were found in finches captured in 2018, a year with much higher precipitation than in 2019. For S. petechia aureola, the only significant difference (p < 0.05) between areas was a wider beak in birds captured in the seasonal forest compared to those from urban areas. Although our sample size does not allow for definitive conclusions, our results provide evidence that the ecology of each species plays a crucial role in shaping their morphological and physiological responses to land use changes and seasonal environmental changes.

The Distribution of Agromyza Species in Nagorno-Karabakh Region and Infestation Rate of Host Plants

Grigoryan, N.M. and V.S. Hovhannisyan. 2024. The Distribution of Agromyza Species in Nagorno-Karabakh Region and Infestation Rate of Host Plants. Arab Journal of Plant Protection, 42(4): 461-464. https://doi.org/10.22268/AJPP-001278 In a survey carried out during the period 2019-2022, two species of Agromyza (Diptera: Agromyzidae) were identified for the first time in Nagorno-Karabakh region: A. parca and A. varifrons. This study focused on these species’ description, distribution, infestation of host plants and determination of some morphometric measurements. High infestation rate (65-70%) was noted on Alopecurus pratensis. Morphometric measurements confirmed the presence of sexual dimorphism in both identified species, especially the male wing length of A. parca in NagornoKarabakh was shorter by 0.7-0.9 mm, and in females 0.3-0.6 mm, whereas the wing length of A. varifrons was shorter by 0.5-0.8 mm. Keywords: Leaf miners, host-plants, morphometric measurements.

Tailless control of a four-winged flapping-wing micro air vehicle with wing twist modulation

This paper describes the tailless control system design of a flapping-wing micro air vehicle in a four-winged configuration, which can provide high control authority to be stable and agile in flight conditions from hovering to maneuvering flights. The tailless control system consists of variable flapping frequency and wing twist modulation. The variable flapping frequency creates rolling moments through differential vertical force from flapping mechanisms that can be independently driven on the left and right sides. The wing twist modulation changes wing tension, resulting in vertical and horizontal force variations during one flap cycle and generating pitching and yaw moments. We presume that the wing geometry and implementation method of wing-root actuation are related to the control authority of wing twist modulation. Then, the control system's performance is analyzed for various wing geometries and implementation methods, including wing length, leading-edge thickness, camber angle, and vein configuration. Furthermore, the cross-coupling effect is examined for the wing twist modulation, and a control surface interconnect is designed to compensate for the decrease of pitch control authority and adverse rolling moment. The refined wing and control mechanism demonstrated its high control authority without significant loss of vertical force and power efficiency. The flight experiments validated that the control system based on wing twist modulation is suitable for four-winged flapping-wing micro air vehicles, providing sufficient control moment and minimizing the cross-coupling effect.

Investigating the Power Extraction of Applying Hybrid Pitching Motion on a Wing with Leading and Trailing Flaps

This research utilized a hybrid trajectory on a wing incorporating a dual flap with the goal of enhancing performance. The hybrid profiles initiate with a non-sinusoidal pattern during the interval 0.0 ≤ t/T ≤ 0.25, evolving toward a sinusoidal pattern within the range 0.25 &lt; t/T ≤ 0.5. Similarly, the hybrid motion follows a non-sinusoidal pattern in the range 0.5 &lt; t/T ≤ 0.75, before shifting back to a sinusoidal pattern within the range 0.75 &lt; t/T ≤ 1.0. The effectiveness of using a hybrid trajectory on a wing with leading and trailing flaps in enhancing the energy harvesting performance is examined through numerical simulations. The results demonstrate that hybrid trajectories applied to a two-flap wing configuration outperform a single flat plate and a wing with leading and trailing flaps both operating under a sinusoidal trajectory. The wing length spans from 45% to 55%, with the leading flap length ranging from 25% to 35%. The trailing flap lengths adjust accordingly to ensure the combined total matches the flat plate’s full length, which is 100%. The wing pitch angle was fixed at 85° while the leading flap’s pitch angle varied between 40° and 55° and the pitch angle of the trailing flap ranged from 0° to 20°. The findings reveal that utilizing hybrid motion on a wing fitted with leading and trailing flaps notably improves power output in comparison to configurations with either one plate or three plates. The power output is achieved at particular dimensions: a leading flap length of 30%, a wing length of 55%, and a trailing flap length of 15%. The corresponding pitch angles are 50° for the leading flap, 85° for the wing, and 10° for the trailing flap. The aforementioned configuration results in a 34.06% increase in output power in comparison to one plate. The maximum efficiency for this setup reaches 44.21%. This underscores the superior performance of hybrid trajectories over sinusoidal trajectories in enhancing energy extraction performance.

Genetic and morphological shifts associated with climate change in a migratory bird

BackgroundRapid morphological change is emerging as a consequence of climate change in many systems. It is intuitive to hypothesize that temporal morphological trends are driven by the same selective pressures that have established well-known ecogeographic patterns over spatial environmental gradients (e.g., Bergman’s and Allen’s rules). However, mechanistic understanding of contemporary morphological shifts is lacking.ResultsWe combine morphological data and whole genome sequencing from a four-decade dataset in the migratory bird hermit thrush (Catharus guttatus) to test whether morphological shifts over time are accompanied by genetic change. Using genome-wide association, we identify alleles associated with body size, bill length, and wing length. Shifts in morphology and concordant shifts in morphology-associated alleles over time would support a genetic basis for the observed changes in morphology over recent decades, potentially an adaptive response to climate change. In our data, bill size decreases were paralleled by genetic shifts in bill size-associated alleles. On the other hand, alleles associated with body size showed no shift in frequency over time.ConclusionsTogether, our results show mixed support for evolutionary explanations of morphological response to climate change. Temporal shifts in alleles associated with bill size support the hypothesis that selection is driving temporal morphological trends. The lack of evidence for genetic shifts in body size alleles could be explained by a large role of plasticity or technical limitations associated with the likely polygenic architecture of body size, or both. Disentangling the mechanisms responsible for observed morphological response to changing environments will be vital for predicting future organismal and population responses to climate change.

Wildfires induce a reduction in body size and morphological variation of an insular endemic insect

Abstract Anthropogenetic disturbance is known to affect population sizes and genetic population structure of many biotas. Wildfires are a major disturbance in many regions of the world, particularly in Mediterranean regions and on the Atlantic islands. Populations of many insects, such as the Madeiran Green Bush‐Cricket (Psalmatophanes barretoi), are threatened by wildfires. However, the effects of wildfires on genetic structure and diversity as well as morphological variation of the populations remain little understood. Therefore, we studied genetic diversity, structure and potential bottlenecks of this species using microsatellites. We also studied morphological variation and fluctuating asymmetry within and between populations to unravel potential effects of wildfires. We did not find any evidence for genetic differentiation of populations, but some populations had high heterozygosity excess, regardless of burning. Morphological variation in burnt areas was lower than in non‐burnt areas. Fluctuating asymmetry of the wing length was significantly higher in burnt areas. Our results show that even genetically well‐connected populations may suffer from bottlenecks leading to reduced morphological variation after disturbance.

Long-term increases in wing length occur independently of changes in climate and climate-driven shifts in body size.

Recent widespread reductions in body size across species have been linked to increasing temperatures; simultaneous increases in wing length relative to body size have been broadly observed but remain unexplained. Size and shape may change independently of one another, or these morphological shifts may be linked, with body size mediating or directly driving the degree to which shape changes. Using hierarchical Bayesian models and a morphological time series of 27 366 specimens from five North American migratory passerine bird species, we tested the roles that climate and body size have played in shifting wing length allometry over four decades. We found that colder temperatures and reduced precipitation during the first year of life were associated with increases in wing length relative to body size but did not explain long-term increases in wing length. We found no conclusive evidence that the slope of the relationship between body size and wing length changed among adult birds in response to any climatic variable or through time, suggesting that body size does not mediate shifts in relative wing length. Together, these findings suggest that long-term increases in wing length are not a compensatory adaptation mediated by size reductions, but rather are driven by non-climatic factors.

Breeding Partners Have Dissimilar Foraging Strategies in a Long-Lived Arctic Seabird.

For long-lived species with biparental care, coordination and compatibility in the foraging behavior of breeding mates may be crucial to successfully raise offspring. While high foraging success is clearly important to reproductive success, it might be equally important that the mate has a complementary foraging strategy. We test whether breeding partners have similar or dissimilar foraging strategies in a species where both partners share breeding responsibilities and exhibit high mate fidelity (thick-billed murre; Uria lomvia). To examine whether thick-billed murres showed complementary in foraging strategies, we attached GPS accelerometers to both partners within 40 thick-billed murre chick-rearing pairs. Individuals within a breeding pair were dissimilar in their foraging trip distance and in their number of dives during foraging trips compared to randomized pairs. Breeding partners were also more similar in wing length than randomized pairs. This result could be related to individual quality as individuals select similar sized partners or select sites that lead to similar sized partners. We conclude that foraging strategy diversity could be maintained in this population either because individuals prefer partners with foraging strategies complementary to their own, or because partners diverge in foraging strategies over multiple breeding season together.

The Interspecific Competition Between Larvae of Aedes aegypti and Major African Malaria Vectors in a Semi-Field System in Tanzania.

Interspecific competition between mosquito larvae may affects adult vectorial capacity, potentially reducing disease transmission. It also influences population dynamics, and cannibalistic and predatory behaviors. However, knowledge of interspecific competition between Ae. aegypti and Anopheles species is limited. The study examined interspecific competition between Ae. aegypti larvae and either An.arabiensis, An. gambiae, or An. funestus on individual fitness in semi-field settings. The experiments involved density combinations of 100:100, 200:0, and 0:200 (Ae. aegypti: Anopheles), reared with and without food, in small habitats (8.5 cm height × 15 cm diameter) with 0.5 liters of water and large habitats (15 cm height × 35 cm diameter) with 1 liter of water. The first group received Tetramin® fish food (0.02 g), while the second group was unfed to assess cannibalism and predation. While interspecific competition affected both genera, Anopheles species experienced greater effects, with reduced survival and delayed development, compared to Ae. aegypti. The mean wing lengths of all species were significantly small in small habitats with a mixed population (p < 0.001). The presence of food reduced cannibalism and predation compared to its absence. These interactions have implications for diseases transmission dynamics and can serve as biological indicators to signal the impacts of vector control interventions.

Performance Analysis in Quantitative Local Ducks in Kediri Regency East Java

The livestock sector is one of the potentials in Indonesia, including duck farming. The duck population in Kediri District in 2022 amounted to 367,987 heads. This research was conducted in 5 subdistricts in Kediri District including Mojo Subdistrict, Ringinrejo Subdistrict, Wates Subdistrict, Ngasem Subdistrict, and Gampengrejo Subdistrict. The variables observed in this research include quantitative traits (femur length, tibia length, shank length, shank circumference, wing length, sternum length, chest width, and chest depth). The differences in the results of this study are influenced by environmental factors and genetics. The conclusion of this study is that the morphometric measurement interval (quantitative traits) of local ducks in each sub-district in Kediri District varies greatly because it is influenced by several factors including the environmental conditions of each sub-district, feed consumption, maintenance methods, and age. The measurement results show that local ducks in Kediri District are still cultivated by farmers and have a variety of sizes. The suggestion of this research is that morphometric measurements (quantitative traits) of local ducks should be carried out in all sub-districts in Kediri District so that it can be known that the body size of ducks in Kediri District is very diverse.

Effect of Thermal Manipulation During Embryogenesis on Pre and Post-Hatch Performance of Stored Hatching Eggs of Japanese Quails

This research investigated the influence of high incubation temperature on hatching, and post-hatch characteristics of stored hatching eggs of Japanese quails. Hatching eggs of Japanese quails were stored for 7 days and incubated under two temperature conditions. The T1 group (control, 75 eggs) was subjected to a standard incubation temperature (37.5°C) while the T2 group (75 eggs) was exposed to a thermal manipulation protocol (of 38.5°C for 5 hours daily between embryonic days 5-15. The egg weight classification, chick weight, chick length, wing length, weekly body traits, body weight, total feed intake, and stress responses, weight of internal organs, whole carcass, breast, neck, wing, thigh, and neck did not significantly differ (P&lt;0.05) between the incubation treatments. Hatchability was higher and early embryonic mortality was lower in T2 than in T1. Late embryonic mortality was lower in T1. Significantly (P≤ 0.05) higher pectoral muscle width at hatch and carcass yield/dressing percentage were observed in quails exposed to thermal manipulation protocol during embryogenesis. It was concluded that exposure of stored eggs to thermal manipulation protocol (of 38.5°C for 5 hours between embryonic days, ED 5-15, T2) during embryogenesis could enhance embryonic and growth traits, as well as carcass traits without any negative effect on stress indicators.

Unpacking WHO and CDC Bottle Bioassay Methods: A Comprehensive Literature Review and Protocol Analysis Revealing Key Outcome Predictors

Background Resistance monitoring is a key element in controlling vector-borne diseases. The World Health Organization (WHO) and Centres for Disease Control and Prevention (CDC) have each developed bottle bioassay methods for determining insecticide susceptibility in mosquito vectors which are used globally. Methods This study aimed to identify variations in bottle bioassay methodologies and assess the potential impact on the data that is generated. Our approach involved a systematic examination of existing literature and protocols from WHO and CDC, with a focus on the specifics of reported methodologies, variation between versions, and reported outcomes. Building on this, we experimentally evaluated the impact of several variables on bioassay results. Results Our literature review exposed a significant inconsistency in the how bioassay methods are reported, hindering reliable interpretation of data and the ability to compare results between studies. The experimental research provided further insight by specifically identifying two key factors that influence the outcomes of bioassays: mosquito dry weight and relative humidity (RH). This finding not only advances our comprehension of these assays but also underscores the importance of establishing precisely defined methodologies for resistance monitoring. The study also demonstrates the importance of controlling bioassay variables, noting the significant influence of wing length, as an indicator of mosquito size, on mortality rates in standardized bioassays. Conclusions Generating data with improved protocol consistency and precision will not only deepen our understanding of resistance patterns but also better inform vector control measures. We call for continued research and collaboration to refine and build consensus on bioassay techniques, to help bolster the global effort against vector-borne diseases like malaria.