Influence of photoperiod on the developmental times of the forensically relevant blow fly species Calliphora vicina (Diptera: Calliphoridae).

Blow flies (Diptera: Calliphoridae) are frequently used in forensic investigations due to their rapid colonization of cadavers. As with other insects, environmental temperature strongly influences their developmental rates. While published research has typically explored not only the impact of the environmental temperature, but also of other factors like tissue type and drug presence on developmental rates, the influence of photoperiod on the developmental rates of forensically relevant blow fly species has remained largely underexplored. Understanding the relationship between photoperiod and developmental times is crucial, as neglecting this aspect could compromise the accuracy of minimum post-mortem interval (minPMI) estimations. The present study investigates the impact of three photoperiod conditions (0:24, 8:16, and 12:12 light:darkness) on the developmental rates of Calliphora vicina, focusing on the duration of the different immature stages and on the total developmental time. Our results revealed significant variation in the intra-puparial stage and total development time across different photoperiods. Notably, a 12:12 photoperiod led to a significantly prolonged intra-puparial stage and total development time compared to the 0:24 photoperiod, suggesting that Calliphora vicina develops faster in total darkness. These findings highlight the importance of considering photoperiod in both laboratory rearing protocols and forensic casework to improve the accuracy and reliability of minPMI estimations. In this regard, preliminary guidelines and recommendations are provided.

Differences in male Aedes aegypti and Aedes albopictus hearing systems facilitate recognition of conspecific female flight tones.

When Aedes albopictus mosquitoes invade regions predominated by Aedes aegypti, either the latter can be displaced or the species can coexist, with potential consequences on disease transmission. Males from both species identify females by listening for her flight sounds. Comparing male hearing systems may provide insight into how hearing could prevent interspecific mating. Here, we show that species-specific differences in female wing beat frequencies are reflected in differences in male ear mechanical tuning frequencies and sound response profiles. Though Aedes albopictus males are attracted to sound, they do not readily display abdominal bending, unlike Aedes aegypti. We observed interspecific differences in male ear mechanical, but not electrical, tuning, suggesting a conserved primary auditory processing pathway. Our work suggests a potential role for hearing in the premating isolation of Aedes aegypti and Aedes albopictus, with implications for predicting future dynamics in their sympatric relationships and our understanding of mosquito acoustic communication.

Intrapuparial stage aging and PMI estimation based on the developmental transcriptomes of forensically important Aldrichina grahami (Diptera: Calliphoridae) gene expression.

Background: The expression profiles of differentially expressed genes (DEGs) during pupal development have been demonstrated to be vital in age estimation of forensic entomological study. Here, using forensically important Aldrichina grahami (Diptera: Calliphoridae), we aimed to explore the potential of intrapuparial stage aging and postmortem interval (PMI) estimation based on characterization of successive developmental transcriptomes and gene expression patterns. Methods: We collected A. grahami pupae at 11 successive intrapuparial stages at 20 °C and used the RNA-seq technique to build the transcriptome profiles of their intrapuparial stages. The DEGs were identified during the different intrapuparial stages using comparative transcriptome analysis. The selected marker DEGs were classified and clustered for intrapuparial stage aging and PMI estimation and then further verified for transcriptome data validation. Ultimately, we categorized the overall gene expression levels as the dependent variable and the age of intrapuparial A. grahami as the independent variable to conduct nonlinear regression analysis. Results: We redefined the intrapuparial stages of A. grahami into five key successive substages (I, II, III, IV, and V), based on the overall gene expression patterns during pupal development. We screened 99 specific time-dependent expressed genes (stage-specific DEGs) to determine the different intrapuparial stages based on comparison of the gene expression levels during the 11 developmental intrapuparial stages of A. grahami. We observed that 55 DEGs showed persistent upregulation during the development of intrapuparial A. grahami. We then selected four DEGs (act79b, act88f, up and ninac) which presented consistent upregulation using RT-qPCR (quantitative real-time PCR) analysis, along with consideration of the maximum fold changes during the pupal development. We conducted nonlinear regression analysis to simulate the calculations of the relationships between the expression levels of the four selected DEGs and the developmental time of intrapuparial A. grahami and constructed fitting curves. The curves demonstrated that act79b and ninac showed continuous relatively increasing levels. Conclusions: This study redefined the intrapuparial stages of A. grahami based on expression profiles of developmental transcriptomes for the first time. The stage-specific DEGs and those with consistent tendencies of expression were found to have potential in age estimation of intrapuparial A. grahami and could be supplementary to a more accurate prediction of PMI.

Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae): A potentially forgotten forensically important species in Malaysia.

Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae) has been collected in human forensic case work. This species is endemic in Malaysia but is not one of the most common species and is often found in outdoor cases. However, it is hypothesized that the presence of L. cuprina larvae may provide useful information in forensic case studies in Malaysia where this species has previously been ignored. This review will discuss the current background knowledge on L. cuprina, particularly when it comes to other forensic cases in Malaysia. General biology as well as key information for forensic work such as geographical distribution and developmental data will be reviewed. Finally, we discuss the potential for L. cuprina to provide beneficial and unique forensic insight into indoor cases with refuse.

Mvolo County, an Onchocerciasis Endemic Area in Western Equatoria State, South Sudan: An Entomological Study to Prepare for a "Slash and Clear" Community-Based Vector Control Intervention.

Background: Mvolo in Western Equatoria of South Sudan has been a hotspot for Onchocerca volvulus transmission since the 1940s. In Mvolo onchocerciasis is a disease of public health importance, associated with onchocerciasis-associated epilepsy including nodding syndrome. Methods: We conducted an entomological study to map the breeding sites of blackflies (Simulium damnosum, sensu lato) on the river Naam, to allow the removal of vegetation from vector breeding sites, the "slash and clear". Three blackfly catching sites were established along the river. Focus group discussions were also conducted to assess the willingness of the communities to support the "slash and clear" intervention and the semi-annual distribution of ivermectin. Results: A total of 2466 female S. damnosum s.l. were caught in 14×11h (06.00-15.00) catches. The highest biting density of 4210.25 flies/month/h and monthly biting rate (MBR) of 11,482.25 bites/man/month were observed in November 2023. Biting density and MBR reduced to zero in the intervention site by April 2024. While the mean parity rate was 31% (CI: 0.2976±0.9176). Two diurnal biting peaks were observed, one from 9:00-10:00 (at the bridge site) and a prominent one from 14:00-15:00 in the two catching sites in Mvolo. Along the river Naam, only one site was found productive for S. damnosum s.l.; and the larvae and adults were morphologically associated with the anthropophilic S. damnosum. The "slash and clear" intervention was implemented at Dogoyabolu along the river Naam. Communities expressed willingness to support a "slash and clear" intervention and the semi-annual distribution of ivermectin. Conclusion: S. damnosum active breeding was identified along the river Naam in a stretch of 3-5 km close to human settlements. Highest blackfly biting density was 4210.25 flies/month/h, and two fly biting peaks were observed. A community "slash and clear" vector control was implemented, and will be prospectively monitored.

Culicoides Latreille (Diptera, Ceratopogonidae) of Colombia: records from the collection of insects of medical importance from National Institute of Health.

Background: The collection of insects of medical importance from the Instituto Nacional de Salud, INS (Bogotá, Colombia: https://www.ins.gov.co/Paginas/Inicio.aspx), was started in 1934 with the aim of being an institutional and national repository of the biodiversity of insects involved in vector-borne diseases of importance in public health. Today, the entomological collection includes more than 7,500 specimens.The ceratopogonid insects are one group of Diptera that are represented in this collection. Within the Ceratopogonidae, the genus Culicoides Latreille, 1809 is relevant in public health because of the nuisance caused by their bites when they are presented in great abundance and because of their role as vectors of several agents (virus, protozoa and nematodes) that cause diseases to humans and to animals (Mellor et al. 2000, Mullen 2002). An overview of the Ceratopogonidae, represented in this collection, is presented here. A total of 801 individuals, mainly adults of the genus Culicoides (90%) are represented. The collection is the result of the effort of several researchers of the Group of Entomology at INS. These researchers collected ceratopogonids when they went to different transmission scenarios of vector-borne diseases in Colombia, with the purpose of making entomological characterisations including the processing, assembly and identification of the specimens in the laboratory. New information: New information about the geographical distribution of 39 species of the genus Culicoides in Colombia. All data have been uploaded to GBIF and are publicly available there.

Field evaluation of female- and male-targeted traps for Ceratitis capitata (Diptera: Tephritidae).

Mediterranean fruit fly (Medfly) Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is a globally significant economic pest for which lure based trapping can be used to monitor established populations and for surveillance. Either female- or male-targeted traps can be used; however, recommendations on which to apply are inconsistent and many programs rely on male-targeted traps. Here, we compare the performance of male-targeted traps (Lynfield Trap with Trimedlure) and female-targeted traps (Biotrap Globe trap with the 3-component lure-TMA Plus) in apple orchards in south-west Western Australia over 2 years (September 2019 to September 2021). Male-targeted traps caught more Medflies overall than female-targeted traps, although the difference was minor. However, female-targeted traps were better at attracting Medfly early in the season when populations were small; and were more likely to capture at least one fly when their paired male-targeted trap caught none. Conversely, male-targeted traps were more likely to capture Medflies late in the season and were more likely to catch high numbers of Medflies. Consequently, female-targeted traps may be better at detecting Medfly early in the season, and male-targeted traps may be better at detecting Medfly abundance late in the season, at least in apple orchards. Our results suggest that either or both trap-types could be used for monitoring Medfly populations, with the optimal solution being dependent on the intended application.

The Mosquito Fauna of Arizona: Species Composition and Public Health Implications.

Arizona is home to many mosquito species, some of which are known vectors of infectious diseases that harm both humans and animals. Here, we provide an overview of the 56 mosquito species that have been identified in the State to date, but also discuss their known feeding preference and the diseases they can (potentially) transmit to humans and animals. This list is unlikely to be complete for several reasons: (i) Arizona's mosquitoes are not systematically surveyed in many areas, (ii) surveillance efforts often target specific species of interest, and (iii) doubts have been raised by one or more scientists about the accuracy of some collection records, which has been noted in this article. There needs to be an integrated and multifaceted surveillance approach that involves entomologists and epidemiologists, but also social scientists, wildlife ecologists, ornithologists, representatives from the agricultural department, and irrigation and drainage districts. This will allow public health officials to (i) monitor changes in current mosquito species diversity and abundance, (ii) monitor the introduction of new or invasive species, (iii) identify locations or specific populations that are more at risk for mosquito-borne diseases, and (iv) effectively guide vector control.

Caste-biased patterns of brain investment in the subterranean termite Reticulitermes flavipes.

Investment into neural tissue is expected to reflect the specific sensory and behavioral capabilities of a particular organism. Termites are eusocial insects that exhibit a caste system in which individuals can develop into one of several morphologically and behaviorally distinct castes. However, it is unclear to what extent these differences between castes are reflected in the anatomy of the brain. To address this question, we used deformation-based morphometry to conduct pairwise comparisons between the brains of different castes in the eastern subterranean termite, Reticulitermes flavipes. Workers exhibited enlargement in the antennal lobes and mushroom bodies, while reproductives showed increased investment into the optic lobes and central body. In addition, caste-specific enlargement was observed in regions that could not be mapped to distinct neuropils, most notably in soldiers. These findings demonstrate a significant influence of caste development on brain anatomy in termites alongside convergence with eusocial hymenopteran systems.

Aseasonal, undirected migration in insects: 'Invisible' but common.

Many insect pests are long-distance migrants, moving from lower latitudes where they overwinter to higher latitudes in spring to exploit superabundant, but seasonally ephemeral, host crops. These seasonal long-distance migration events are relatively easy to recognize, and justifiably garner much research attention. Evidence indicates several pest species that overwinter in diapause, and thus inhabit a year-round range, also engage in migratory flight, which is somewhat "invisible" because displacement is nondirectional and terminates among conspecifics. Support for aseasonal, undirected migration is related to recognizing true migratory flight behavior, which differs fundamentally from most other kinds of flight in that it is nonappetitive. Migrating adults are not searching for resources and migratory flight is not arrested by encounters with potential resources. The population-level consequence of aseasonal, undirected migration is spatial mixing of individuals within the larger metapopulation, which has important implications for population dynamics, gene flow, pest management, and insect resistance management.

Population-specific thermal responses contribute to regional variability in arbovirus transmission with changing climates.

Temperature is increasing globally, and vector-borne diseases are particularly responsive to such increases. While it is known that temperature influences mosquito life history traits, transmission models have not historically considered population-specific effects of temperature. We assessed the interaction between Culex pipiens population and temperature in New York State (NYS) and utilized novel empirical data to inform predictive models of West Nile virus (WNV) transmission. Genetically and regionally distinct populations from NYS were reared at various temperatures, and life history traits were monitored and used to inform trait-based models. Variation in Cx. pipiens life history traits and population-dependent thermal responses account for a predicted 2.9°C difference in peak transmission that is reflected in regional differences in WNV prevalence. We additionally identified genetic signatures that may contribute to distinct thermal responses. Together, these data demonstrate how population variation contributes to significant geographic variability in arbovirus transmission with changing climates.

The genetic basis of variation in Drosophila melanogaster mating behavior.

Mating behavior is an essential fitness trait. We used the inbred, sequenced lines of the Drosophila Genetic Reference Panel (DGRP) to gain insights into the evolution of mating success and to evaluate the overlap in genetic architecture of mating behavior between the sexes. We found significant genetic variation for mating success when DGRP males and females from the same line were mated together, and when DGRP males and females were mated to an unrelated strain of the opposite sex. The mating success of DGRP males and females was not correlated when they were paired with the unrelated strain, suggesting independent genetic architecture of mating success in males and females that was confirmed by genome-wide association analyses. However, the mating success between pairs of the same or different DGRP lines was predicted accurately by the respective female and male mating success with the unrelated line.

A geometric morphometric analysis of wing variations in shape and size of the blue bottle fly, Calliphora vicina (Diptera: Calliphoridae).

A geometric morphometric analysis was performed on the right wing of adult Calliphora vicina (Robineau-Desvoidy) collected across 4 altitudinal levels in Sicily. The objective of this study was to assess differences in shape and centroid size (CS) between females and males and across elevations. The wings analyzed in this study were removed from adults of C. vicina collected with baited traps at 20, 700, 1,153, and 1,552; for this study, 19 landmarks were identified in each wing. The coordinates of the landmarks were aligned and superimposed to prevent variations due to position, orientation, and scale; they were then scaled to the same CS and recentered. CS and Procrustes differences were, respectively, used to assess variations in size and shape. Significant differences were observed in wing shape between males and females but not between all altitudinal levels. Female wings were found to be significantly larger than males (P < 0.01). Wings of flies collected at the highest altitudinal level resulted in significantly larger wings than those collected at lower altitudes (P < 0.001), with CS values ranging from 12.1 to 14.1. Variation in wing shape can impact thermal regulation, and therefore, oxygen content, temperature, atmospheric pressure, and solar radiation can have an effect on an insect's body and activity levels. At high elevations and lower temperatures, larger wings could mean less energy expenditure when flying to increase body temperature.

Photoreceptors for immediate effects of light on circadian behavior.

Animals need to sharpen their behavioral output in order to adapt to a variable environment. Hereby, light is one of the most pivotal environmental signals and thus behavioral plasticity in response to light can be observed in diurnal animals, including humans. Furthermore, light is the main entraining signal of the clock, yet immediate effects of light enhance or overwrite circadian output and thereby mask circadian behavior. In Drosophila, such masking effects are most evident as a lights-on response in two behavioral rhythms - the emergence of the adult insect from the pupa, called eclosion, and the diurnal rhythm of locomotor activity. Here, we show that the immediate effect of light on eclosion depends solely on R8 photoreceptors of the eyes. In contrast, the increase in activity by light at night is triggered by different cells and organs that seem to compensate for the loss of each other, potentially to ensure behavioral plasticity.

Comparing Entomology-Themed Outreach Events: Annual Festivals and Open Houses in the United States.

Over the past several decades, there has been a growing prevalence of entomology-themed outreach events, which seek to educate the public about insects and other arthropods, fostering a greater appreciation and understanding of these often misunderstood organisms. In 2016, a comparative analysis was initiated to identify science institutions across the United States engaged in providing annual entomology-themed outreach events to the public. Utilizing literature reviews and online searches, several science institutions were identified and subsequently contacted to partake in a survey regarding their events. The survey received a response rate of 93%. Results from the survey offered valuable insights into these entomology-themed outreach events, including records of attendance, event structures, funding sources, popular exhibits, and the impacts on attendees, hosting institutions, and local communities. While the majority of these events have remained in place since the survey, many have experienced significant disruptions due to the COVID-19 pandemic, prompting some to adapt to innovative online formats and virtual experiences. Despite these challenges, the commitment to entomological outreach continues today, highlighting the resilience and adaptability of the entomology community.

Discovery of the Larvae and Pupae of the Black Fly Simulium (Gomphostilbia) khelangense and Breeding Habitats of Potential Pest Species of the S. (G.) chumpornense Subgroup (Simuliidae).

Two species of black flies (Simuliidae) in Thailand, Simulium chumpornense Takaoka and Kuvangkadilok, 2000, and S. khelangense Takaoka, Srisuka & Saeung, 2022, are potent vectors of avian blood protozoa of the genera Leucocytozoon and Trypanosoma and are pests of domestic avian species. Although the adults are abundant throughout Thailand, information on their breeding habitats is limited, and the immature stages of S. khelangense are unknown. We collected the larvae and pupae of S. khelangense from the Mekong River, the first-ever record of Simuliidae from this large continental river. Mitochondrial cytochrome c oxidase I and internal transcribed spacer 2 were used to associate the larvae and pupae with known adults. Both genetic markers strongly supported their identity as S. khelangense. The larvae and pupa of S. khelangense are described. The pupal gill filaments, larval abdominal protuberances, and setae distinguish this species from other members of the S. varicorne species group. The immature stages of S. chumpornense inhabit a wide variety of flowing waters, from small streams (3 m wide) to enormous continental rivers (400 m wide); thus, S. chumpornense is a habitat generalist. In contrast, S. khelangense was found only in the large Mekong River and is, therefore, a habitat specialist. Both species can exploit their principal habitats and produce abundant adult populations.

Molecular differentiation analysis of ten putative species of Fannia (Diptera: Fanniidae) collected in carrion-baited traps from Colombia.

The genus Fannia is the most representative of the Fannidae family of true flies with worldwide distribution. Some species are attracted to decomposing materials and live vertebrate animals, which makes them important in forensics, medical and veterinary fields. However, identifying Fannia species can be difficult due to the high similarity in the external morphology of females and limited descriptions and morphological keys. Herein, molecular markers could provide a complementary tool for species identification. However, molecular identification has still limited application since databases contain few data for neotropical species of Fannia. This study assessed the potential of two molecular markers, the COI-3' region and internal transcribed spacer 2 (ITS2), to differentiate 10 putative species of the genus Fannia from Colombia using distance-based and tree-based approaches. The partial ITS2 and/or COI-3' regions allowed molecular diagnosis of six species, while pairs of species Fannia colazorrensis + F. dodgei and F. laclara + F. aburrae are conflicting. Although these results might suggest that conflicting pair species are conspecific, consistent morphological differences between males do not support this hypothesis. The lack of differentiation at the nuclear and mitochondrial molecular markers for the conflicting species may be due to incomplete evolutionary lineage separation, hybridization, or introgression events. In addition, sexual selection on male morphological traits before species-specific differences in molecular markers emerge may partially explain the results. Our study provides a valuable dataset to identify and confirm some Fannia species molecularly. Further, they could be used to associate females and immature stages with their conspecifics as a baseline to deep into their biology, ecology, distribution and potential applications in forensic and medico-veterinary entomology.

Mosquito species identity matters: unraveling the complex interplay in vector-borne diseases.

BACKGROUND: Research on vector-borne diseases has traditionally centred on a limited number of vertebrate hosts and their associated pathogens, often neglecting the broader array of vectors within communities. Mosquitoes, with their vast species diversity, hold a central role in disease transmission, yet their capacity to transmit specific pathogens varies considerably among species. Quantitative modelling of mosquito-borne diseases is essential for understanding transmission dynamics and requires the necessity of incorporating the identity of vector species into these models. Consequently, understanding the role of different species of mosquitoes in modelling vector-borne diseases is crucial for comprehending pathogen amplification and spill-over into humans. This comprehensive overview highlights the importance of considering mosquito identity and emphasises the essential need for targeted research efforts to gain a complete understanding of vector-pathogen specificity. METHODS: Leveraging the recently published book, 'Mosquitoes of the World', I identified 19 target mosquito species in Europe, highlighting the diverse transmission patterns exhibited by different vector species and the presence of 135 medically important pathogens. RESULTS: The review delves into the complexities of vector-pathogen interactions, with a focus on specialist and generalist strategies. Furthermore, I discuss the importance of using appropriate diversity indices and the challenges associated with the identification of correct indices. CONCLUSIONS: Given that the diversity and relative abundance of key species within a community significantly impact disease risk, comprehending the implications of mosquito diversity in pathogen transmission at a fine scale is crucial for advancing the management and surveillance of mosquito-borne diseases.

Demographic inference from the mt-DNA COI gene and wing geometry of Culex gelidus (Diptera: Culicidae), an important vector of Japanese encephalitis in Thailand.

Culex gelidus (Diptera: Culicidae), an important vector of the Japanese encephalitis virus (JEV), contributes to human viral encephalitis in many Asian countries, including Thailand. This study represents the first investigation of the demographic patterns of Cx. gelidus populations in Thailand using cytochrome c oxidase subunit I (COI) gene analysis and wing geometric morphometrics (GM). Mosquitoes were collected from 10 provinces across six regions of Thailand in 2022. Analysis of the COI sequences (n = 182) indicated high haplotype diversity (0.882) and low nucleotide diversity (0.006), with 72 haplotypes identified. The haplotype network demonstrated no profound splits among the geographic populations. Neutral tests, including Tajima's D and Fu's Fs, displayed negative values, with a significant result observed for Fu's Fs (-33.048, p < 0.05). The mismatch distribution analysis indicated that the population does not statistically deviate from a model of sudden population expansion (SSD = 0.010, p > 0.05; Rg = 0.022, p > 0.05). The estimations suggest that the Cx. gelidus population in Thailand began its expansion approximately between 459,243 and 707,011 years ago. The Mantel test showed no significant relationship between genetic and geographic distances (r = 0.048, p > 0.05). Significant phenotypic differences (based on wing shape) were observed among most populations. Additionally, in this study, we found no significant relationships between phenotypic and genetic distances (r = 0.250, p > 0.05). Understanding the genetic and morphological dynamics of Cx. gelidus is vital for developing targeted surveillance and vector control measures. This knowledge will also help to predict how future environmental changes might affect these populations, thereby informing long-term vector management strategies.

An illustrated identification key to early instar larvae of forensically important Muscidae (Diptera) of the western Palaearctic region.

There is a significant gap in the availability of comprehensive identification keys for the early larval stages of forensically important fly species. While well-documented identification keys exist for the third instar larvae, particularly for the Calliphoridae, Muscidae and Sarcophagidae families, there is a notable scarcity of keys for the first, except Calliphoridae, and the second instar larvae, with no such resources available for muscid species. The second instar larvae suffer the most from the lack of morphological descriptions and available identification keys. The Muscidae is one of the most frequently reported dipteran families of forensic importance colonising animal cadavers and human corpses. Nevertheless, descriptions of the morphology of their early instars remain scarce and limited to only a few species, thus their larval identification is challenging or impossible. Considering the numerous challenges associated with studying small-sized entomological material, we tested whether it is feasible to identify muscid flies to the species or at least genus level based predominantly on the details of the cephaloskeleton. To overcome the obstacle of observing details of small sclerites, especially their shapes and interconnections, we effectively employed confocal laser scanning microscopy (CLSM) as a supplementary method for light microscopy (LM). This study provides an identification key for first and second instar larvae of forensically important muscid species from the western Palaearctic (Europe, North Africa, Middle East). The proposed key primarily utilises details of the cephaloskeleton with only addition of external morphology.