Cuticular hydrocarbons (CHCs) are ubiquitous among insects where they form an outer wax layer that helps maintain water balance and prevent desiccation. In social insects, CHCs were subsequently co-opted as semiochemicals in many contexts, including nestmate recognition, which maintains boundaries among competing colonies by ousting non-nestmates. In some ant populations, workers do not discriminate against non-nestmates. This leads to the development of supercolonies, a large network of interconnected nests exchanging unrelated individuals. In this study, we investigate CHC production by workers and their resistance to desiccation in the ant Nylanderia fulva, which exhibits supercolonial behavior within its invasive range in the USA. We found greatly reduced CHC production by workers and increased susceptibility toward desiccation compared to other invasive ants of similar body size. This relative absence of CHCs sheds light on the susceptibility of this species to abiotic stress through desiccation with implications for its potential distribution and its development of large supercolonies in its invasive range by impairing nestmate recognition.

Most population genetic studies concern spatial genetic differentiation, but far fewer aim at analyzing the temporal genetic changes that occur within populations. Vector species, including mosquitoes and biting midges, are often characterized by oscillating adult population densities, which may affect their dispersal, selection, and genetic diversity over time. Here, we used a population of Culicoides sonorensis from a single site in California to investigate short-term (intra-annual) and long-term (inter-annual) temporal variation in genetic diversity over a 3 yr period. This biting midge species is the primary vector of several viruses affecting both wildlife and livestock, thus a better understanding of the population dynamics of this species can help inform epidemiological studies. We found no significant genetic differentiation between months or years, and no correlation between adult populations and the inbreeding coefficient (F-IS). However, we show that repeated periods of low adult abundance during cooler winter months resulted in recurring bottleneck events. Interestingly, we also found a high number of private and rare alleles, which suggests both a large, stable population, as well as a constant influx of migrants from nearby populations. Overall, we showed that the high number of migrants maintains a high level of genetic diversity by introducing new alleles, while this increased diversity is counterbalanced by recurrent bottleneck events potentially purging unfit alleles each year. These results highlight the temporal influences on population structure and genetic diversity in C. sonorensis and provide insight into factors effecting genetic variation that may occur in other vector species with fluctuating populations.

BACKGROUND: A frequent goal of pest management strategies targeting social insects is total colony elimination. Insecticidal baits are highly effective at controlling social insect pests, although their ability to provide total colony elimination has only been well studied in a few species. Genetically testing colony elimination in many urban pest ants can be challenging due to indistinct colony boundaries observed in unicolonial, invasive species; however, some pest ants, such as the dark rover ant (Bra-chymyrmex patagonicus), maintain strict colony borders through aggression towards non-nestmates. Each of these distinct colonies can be identified using molecular markers, allowing for the tracking of individual colonies pre- and post-treatment to measure colony density. While counting the number of foraging workers to assess treatment efficacy may suffice in some cases, it offers little insight into the colony-level impacts of a treatment. RESULTS: Using microsatellite markers, distinct rover ant colonies were identified and tracked around residential structures before and after the application of an imidacloprid bait. The number of foraging ants at the treated structures was reduced by an average of 83.0% over a 28-day observation period. Baiting also significantly reduced the total number of colonies present. At the treatment structures, only similar to 25% of the original colonies remained at the end of the study. Colonies with foraging trails <1.5 m from a bait station had a higher chance of being eliminated. CONCLUSION: Using insecticidal baits against B. patagonicus can be highly effective at colony elimination; however, with such small foraging ranges and high colony densities, proper placement is required to ensure enough bait is properly positioned to treat all colonies affecting a structure. (C) 2022 Society of Chemical Industry.

Social insect colonies are characterized by an efficient division of labor, allowing high-value individuals (i.e., reproductives and brood) to be sheltered from tasks associated with increased risk of pathogen exposure, such as foraging or corpse disposal. This social organization helps limit the transmission of disease throughout the colony. Further, individuals can actively respond to imminent disease threats by altering their behaviors as a means of social immunity. In subterranean termites, although workers typically avoid detected pathogens, they can be attracted to pathogen cues when a nestmate is infected. Infected termites are usually groomed, but they may instead be cannibalized if the infection has already become lethal. The mechanisms governing these changes in behavior are unclear. We set out to examine immediate changes in individual behaviors, investigating the role that the infected individual plays in communicating its infection status to nestmates. We also assessed gradual changes in social organization after the re-introduction of an infected termite to the colony. Our results reveal that infected termites likely do not signal their infection status to nestmates through shaking behaviors and reduced movements, suggesting the occurrence of other mechanisms used in communicating infection. We also found that infected termites do not self-isolate and may travel to the densest part of the colony, where they can potentially benefit from grooming by large groups of nestmates. These results provide new insights into how individual changes in immune behaviors contribute to overall colony health, highlighting that, at early stages of infection, termites favor a rescuing strategy rather than isolation and/or cannibalization.

In insects, ecological competition has often resulted in phenotypic changes and modifications to foraging areas. In termites - and social insects as a whole - colonies cannot easily escape competition through the relocation of their colony. In these species, the outcomes of inter and intra-specific competition are influenced by different life history traits, such as colony size, breeding system (number and types of reproductives), food preference, tunneling patterns, nest site selection, and antagonism between colonies. Here, we investigated variation in breeding system and spatial distribution among colonies of a higher termite Amitermes parvulus and a subterranean termite Reticulitermes flavipes within an urban landscape. We first developed microsatellite markers as a tool to study these life history traits in A. parvulus . Second, we assessed competitive exclusion or tolerance of A. parvulus and R. flavipes colonies by determining their fine-scale distribution using monitoring stations on a grid site, and their large-scale distribution across an urban landscape. Third, we investigated the breeding system of A. parvulus colonies. We showed that the numerous colonies of R. flavipes inhabiting a restricted area contrast with the few, but spatially expansive colonies of A. parvulus , suggesting these species face different degrees of intra-specific competition. We showed that colonies of A. parvulus frequently merged together, and all of them were headed by inbred neotenic reproductives, two characteristics rarely observed in higher termites. Overall, our study revealed drastic differences in colony structure, breeding systems and foraging ranges between the two species. These differences may reflect differences in food preference and food availability between the two species allowing their co-existence within the same urban environment.

Summary After humans, social insects represent one of the most complex groups of social organisms, relying on a well-organized communication system among colony members. The transfer of information among individuals is primarily based on cuticular hydrocarbons (CHC). These chemical compounds, produced by all insects, initially evolved to prevent water loss 1 . They were subsequently co-opted as semiochemicals to communicate various types of information. This includes nestmate recognition in social insects 2,3 , enabling different colonies to partition resources by ousting conspecific competitors. In this study, we report the near complete loss of CHC production by workers of the ant Nylanderia fulva . This absence of CHCs is a double-edged sword. It represents a causative agent in the ecological success of this ant species — enabling the development of a large supercolony in its invasive range through limited ability to differentiate nestmates— but increases the risk of suffering ecological stress through desiccation.

Proper vector surveillance relies on the ability to identify species of interest accurately and efficiently, though this can be difficult in groups containing cryptic species. Culicoides Latreille is a genus of small biting flies responsible for the transmission of numerous pathogens to a multitude of vertebrates. Regarding pathogen transmission, the C. variipennis species complex is of particular interest in North America. Of the six species within this group, only C. sonorensis Wirth & Jones is a proven vector of bluetongue virus and epizootic hemorrhagic disease virus. Unfortunately, subtle morphological differences, cryptic species, and mitonuclear discordance make species identification in the C. variipennis complex challenging. Recently, single-nucleotide polymorphism (SNP) analysis enabled discrimination between the species of this group; however, this demanding approach is not practical for vector surveillance. The aim of the current study was to develop a reliable and affordable way of distinguishing between the species within the C. variipennis complex, especially C. sonorensis. Twenty-five putative microsatellite markers were identified using the C. sonorensis genome and tested for amplification within five species of the C. variipennis complex. Machine learning was then used to determine which markers best explain the genetic differentiation between species. This led to the development of a subset of four and seven markers, which were also tested for species differentiation. A total of 21 microsatellite markers were successfully amplified in the species tested. Clustering analyses of all of these markers recovered the same species-level identification as the previous SNP data. Additionally, the subset of seven markers was equally capable of accurately distinguishing between the members of the C. variipennis complex as the 21 microsatellite markers. Finally, one microsatellite marker (C508) was found to be species-specific, only amplifying in the vector species C. sonorensis among the samples tested. These microsatellites provide an affordable way to distinguish between the sibling species of the C. variipennis complex and could lead to a better understanding of the species dynamics within this group. Additionally, after further testing, marker C508 may allow for the identification of C. sonorensis with a single-tube assay, potentially providing a powerful new tool for vector surveillance in North America.

Abstract The ecological success of ants has made them abundant in most environments, yet inter‐ and intraspecific competition usually limit nest density for a given population. Most invasive ant populations circumvent this limitation through a supercolonial structure, eliminating intraspecific competition through a loss of nestmate recognition and lack of aggression toward non‐nestmates. Native to South America, Brachymyrmex patagonicus has recently invaded many locations worldwide, with invasive populations described as extremely large and dense. Yet, in contrast with most invasive ants, this species exhibits a multicolonial structure, whereby each colony occupies a single nest. Here, we investigated the interplay between genetic diversity, chemical recognition, and aggressive behaviors in an invasive population of B. patagonicus. We found that, in its invasive range, this species reaches a high nest density with individual colonies located every 2.5 m and that colony boundaries are maintained through aggression toward non‐nestmates. This recognition and antagonism toward non‐nestmates is mediated by chemical differentiation between colonies, as different colonies exhibit distinct chemical profiles. We highlighted that the level of aggression between colonies is correlated with their degree of genetic difference, but not their overall chemical differentiation. This may suggest that only a few chemical compounds influence nestmate recognition in this species or that weak chemical differences are sufficient to elicit aggression. Overall, this study demonstrates that invasive ant populations can reach high densities despite a multicolonial structure with strong aggression between colonies, raising questions about the factors underlying their ecological success and mitigating negative consequences of competitive interactions.

Background: Proper vector surveillance relies on the ability to identify species of interest accurately and efficiently, though this can be difficult in groups containing cryptic species. Culicoides is a genus of small biting flies responsible for the transmission of numerous pathogens to a multitude of vertebrates. Regarding pathogen transmission, the C. variipennis species complex is of particular interest in North America. Of the six species within this group, only C. sonorensis is a proven vector of bluetongue virus and epizootic hemorrhagic disease virus. Unfortunately, subtle morphological differences, cryptic species, and mitonuclear discordance make species identification in the C. variipennis complex challenging. Recently, a SNP analysis enabled discrimination between the species of this group; however, this demanding approach is not practical for vector surveillance. Methods: The aim of the current study was to develop a reliable and affordable way of differentiating the species within the C. variipennis complex, especially C. sonorensis . Twenty-five putative microsatellite markers were identified using the C. sonorensis genome and tested for amplification within five species of the C. variipennis complex. Machine learning was then used to determine which markers best explain the genetic differentiation between species. This led to the development of a subset of four and seven markers which were also tested for species differentiation. Results: A total of 21 microsatellite markers were successfully amplified in the species tested. Clustering analyses of all of these markers recover the same species-level identification as the previous SNP data. Additionally, the subset of seven markers was equally capable of accurately differentiating the members of the C. variipennis complex as the 21 microsatellite markers. Finally, one microsatellite marker ( C508 ) was found to be species-specific, only amplifying in the vector species C. sonorensis among the samples tested. Conclusions: These microsatellites provide an affordable way in which to differentiate the species of the C. variipennis complex and could lead to a better understanding of the species dynamics within this group. Additionally, after further testing, marker C508 may allow for the identification of C. sonorensis with a single-tube assay, potentially providing a powerful new tool for vector surveillance in North America.

Sexually antagonistic selection (SAS) occurs when distinct alleles are differentially selected in each sex. In the invasive tawny crazy ant, Nylanderia fulva, a genomic region is under SAS, while the rest of the genome is randomly selected in males and females. In this study, we designed a suite of 15 microsatellite markers to study the origin and evolution of SAS in N. fulva. These SAS markers were polymorphic, with allelic frequencies that are highly different between males and females. All haploid males carry only a subset of the alleles present in the population, while females are reliably heterozygous, with one allele from the male gene pool and a different allele inherited from their mother. In addition, we identified six polymorphic markers not associated with SAS and six markers yielding consistent, yet monomorphic, amplification in the introduced range of this species. Reaction condition optimizations allowed all retained markers to be co-amplified in four PCR mixes. The SAS markers may be used to test for the strength and the extent of the genomic regions under SAS in both the native and introduced ranges of N. fulva, while the set of non-SAS loci may be used to assess the invasion route of this species. Overall, the application of these microsatellite markers will yield insights into the origin and evolution of SAS within and among species of the genus Nylanderia.

Atlantic blue crabs (Callinectes sapidus) are ecologically and commercially fundamental. Life stages are punctuated with migration. Adults and juveniles live in estuaries and sounds. Larval stages develop in the coastal ocean. Juvenile and adult crabs occupy habitats from high salinities to fresh water. We determined whether maturing juvenile and adult blue crab habitat use is reflected in mitochondrial cytochrome oxidase 1 haplotypes. High salinity crabs had lower haplotype diversity (0.7260 ± .03900) compared to spawning crabs (0.9841 ± .00021) and low salinity crabs (0.94154 ± .00118). Significant pairwise differences in haplotypes were found between high salinity and spawning crabs (Nm = 0.26018, p < 0.001), and between high salinity and low salinity crabs (Nm = 0.19482, p < 0.001) indicating a lack of gene flow. Crabs from high salinity had highly significant genetic differentiation compared to spawning crabs (Fst = 0.11830, p < 0.001) and low salinity crabs (Fst = 0.09689, p < 0.001). Results support the hypothesis that genetics influence habitat selection. Crab larvae mix in the coastal ocean but occupy specific habitats upon return to sounds and estuaries. These findings have implications for the management of fisheries.