Parasitology

Neoehrlichia mikurensis is widely detected in tick populations across Europe, yet its host range beyond small rodents remains poorly understood. Large wild mammals are frequently exposed to tick vectors, but their role in the transmission cycle of this bacterium is unclear. Wild boars (Sus scrofa) are among the most abundant large mammals in southern Sweden and commonly inhabit tick-infested environments, making them a potentially relevant host species. In this study, spleen samples from 87 wild boars collected between 2023 and 2025 across southern and central Sweden were analysed for the presence of N. mikurensis DNA using quantitative polymerase chain reaction. N. mikurensis DNA was not detected in any of the analysed samples, despite sampling in regions where the bacterium is known to be prevalent in ticks. These results indicate that wild boars may have a limited role in the maintenance of N. mikurensis in Sweden, although the possibility of rare or transient infection cannot be excluded. Despite being based on negative findings, this study contributes to a more comprehensive understanding of the host range of N. mikurensis and highlights the importance of continued evaluation of wildlife species in the transmission dynamics of tick-borne pathogens.

Feathers incorporated into bird nests are thought to provide multiple benefits, including thermal insulation and protection against nest-dwelling ectoparasites. For instance, in Swallows, it has been suggested that feathers influence infestations by Protocalliphora blow flies either by acting as a physical barrier between larvae and nestlings or through chemical compounds associated with preen oil. However, empirical evidence for an antiparasitic effect of feathers remains equivocal. Using 1,100 naturally infested Tree Swallow (Tachycineta bicolor) nests collected from a nest-box study system monitored in southern Quebec, Canada, between 2004 and 2018, we tested whether nest mass and feather abundance influenced hematophagous ectoparasites (Protocalliphora spp.) loads. We further assessed whether feather abundance directly affected body size using measurements of 1,513 P. sialia puparia, and emergence success using data from 961 Protocalliphora spp. puparia. Nest mass was positively associated with feather abundance, and both variables showed very weak positive relationships with parasite abundance. Feather abundance was unrelated to puparium size nor parasite emergence success. Our results provide no support for an anti-parasitic effect of feather lining within Tree Swallow nests. Using one of the largest datasets assembled to date on feather-lined nests and avian blow flies, we found no evidence that feather abundance impairs Protocalliphora density, development or emergence success. However, our findings do not preclude potential effects of feathers on other ectoparasites or microorganisms with different life histories. Future studies simultaneously monitoring multiple parasite taxa (such as fleas, mites, or bacteria) may therefore be needed to fully evaluate the ecological consequences of feather lining within avian nests.

Land use change is widely recognized as a major driver of emerging zoonotic diseases. Yet, while forest plantations are expanding worldwide, their consequences on the circulation of infectious agents in wildlife remain poorly understood. We investigated how native pine plantations and broadleaved habitats shape rodent-associated infectious hazard in the generalist wood mouse (Apodemus sylvaticus), in southwestern France. Using a multi-season survey, we screened 424 wood mice for a broad range of infectious taxa and compared infection probability and infected-host abundance among pine plantation edges, broadleaved hedgerows embedded within pine plantations and broadleaved forests. Using a combination of linear mixed model and joint species distribution modelling, we tested whether infectious hazard differed between pine plantations and broadleaved forests, whether broadleaved hedgerows shifted these patterns towards those observed in broadleaved forests, and how these habitat-related patterns varied across seasons. We detected 22 potentially infectious taxa, including nine recognized zoonotic bacteria. Season strongly influenced infectious patterns, while habitat effects were taxa and season dependent. In spring, pine plantation edges increased infection probability for Bartonella grahamii and Neoerhlichia mikurensis compared to broadleaved forest, but their strong negative effect on wood mouse abundance resulted in consistently lower infected-host abundance for most infectious taxa. Broadleaved hedgerows showed distinct and season-dependent patterns with infectious profiles resembling those of pine edges in spring, but with higher infectious hazard than both other habitats for several taxa in autumn. Our results highlight that habitat effects on infectious hazard cannot be inferred from infection probability alone. Pine plantations may reduce rodent-associated zoonotic hazard through their strong negative effect on host abundance, despite increasing infection probability for some agents. Forest diversification and restoration initiatives, such as broadleaved hedgerows here, may have unforeseen epidemiological consequences, underscoring the need to integrate disease ecology into forest management and restoration to anticipate variation in zoonotic risks, while maximizing ecological benefits.

Traditional veterinary pathology investigations on wild animals generally conclude a primary cause of death based on a combination of observed clinical signs, gross and microscopic lesions, microbial culture and PCR. However, these methods may fail to capture the broader microbial community of the host. We employed metatranscriptomic sequencing to characterise the potential pathobiome of necropsied marine turtles, comparing these findings with corresponding veterinary pathology reports. Tissue samples were collected from 76 individuals spanning three species of marine turtles -- the green turtle (Chelonia mydas), the hawksbill turtle (Eretmochelys imbricata), and the loggerhead turtle (Caretta caretta) -- across the coastline of New South Wales, Australia. Reported causes of mortality ranged from infectious disease to trauma. We identified a diverse range of parasites within the genera Neospirochis, Hapalotrema, Paraorgymatobothrium, Eimeria and Callitetrarhynchus, and bacteria including the genera Aeromonas, Clostridium, and Vibrio. Novel viruses of the families Astroviridae, Caliciviridae, Hepaciviridae, Hepeviridae, Nyamivirdae, Orthomyxoviridae, Picobirnaviridae and Rhabdoviridae were also identified. In cases where infectious disease was noted in the pathology report, we frequently observed high pathogen read counts consistent with the listed primary diagnoses. However, we also detected a range of incidental infections involving eukaryotic parasites, bacteria, and viruses. The diversity of infectious agents detected in marine turtles suggests that the observed disease outcomes are unlikely to be driven by a single causative agent, but rather by complex interactions between host health, the pathobiome, and environmental stressors. We discuss the potential implications for turtle health and conservation.

Stink bugs (Hemiptera Pentatomidae) are major agricultural pests worldwide, including both native and invasive species. In France, for instance, Halyomorpha halys has rapidly expanded its range, affecting key crops such as hazelnuts, apples, and vegetables. Meanwhile, several native, polyphagous species - Nezara viridula, Palomena prasina and, to a lesser extent, Graphosoma italicum - are re-emerging and impacting diverse crops because of changes in agricultural practices and climatic conditions. Egg parasitoids represent promising biological control agents for these pests, be it through conservation, augmentation or introduction biological control. However, their actual potential is not precisely documented due to a lack of ambitious surveys and possible taxonomic issues. This study aims to address this gap by compiling results obtained from various of egg parasitism surveys of Pentatomidae in France and implementing a large-scale DNA Barcoding approach (2000 individuals) for the emerged parasitoids. Our results indicate a high diversity of egg parasitoids, and detected some taxonomic issues and molecular inconsistencies that warrant further study. Among the three families of egg parasitoids recovered (Encyrtidae, Eupelmide, and Scelionidae), Scelionidae was the most abundant and diverse with nine species of Trissolcus (including the two adventive species, Tr. japonicus and Tr. mitsukurii) and four Telenomus species (including two unidentified to species). Scelionidae dominated the parasitoid assemblages associated with most of the Pentatomidae host species, except for H. halys for which the eupelmid Anastatus bifasciatus was the most common species.

Mycobacterium bovis causes tuberculosis (TB) in cattle, badgers (Meles meles), other wildlife, and humans, posing major animal and public health challenges in Britain and Ireland. Shared genotypes between cattle and badgers have been reported at regional scales, but whether this concordance holds at finer spatial scales, where control interventions operate, and whether it varies by genotype combination, remains unknown. Resolving this is critical for interpreting cross-species transmission risk and designing targeted control measures. We analyzed 1,248 M. bovis isolates from cattle and badgers (Northern Ireland, 1986-2018), including samples from a small, densely sampled intervention research area, using Multi-Locus Variable Number of Tandem Repeats Analysis (MLVA) and kernel discriminant analysis (KDA) across multiple spatial scales (25-95% density contours). We identified 36 MLVA types with four shared between species and one (MLV06) endemic to the area. Spatial concordance varied by type combination and scale: MLVA004-MLVA297 showed strong, bidirectional, scale-robust concordance; MLVA006-MLVA297 was concordant only at fine spatial scales; MLVA004-MLVA006 showed weak, asymmetric concordance; and no concordance was detected when all three types were analysed together. Our findings reveal spatial concordance between shared M. bovis genotypes varied by genotype combination and scale, indicating genotype-specific dynamics within this multi-host system.

In sub-Saharan Africa, human and animal health is threatened by African trypanosomoses. These diseases are caused by trypanosomes that are transmitted primarily by tsetse flies. Vector control remains one of the most effective strategies for reducing disease transmission, but requires a minimum level of knowledge of the population biology, ecology and dispersal patterns of the targeted vectors. In this respect, population genetic approaches using appropriate polymorphic genetic markers can provide valuable tools to inform and optimize control strategies. Senegal is currently engaged in a national tsetse control programme and has been identified as a study country within a European Union-funded research initiative, COMBAT (controlling and progressively minimizing the burden of animal trypanosomosis). Baseline entomological studies are currently underway in the Sine Saloum (Southern Senegal). This preliminary study aimed to characterize the population genetic structure and connectivity of tsetse fly species present in Southern Senegal, Glossina palpalis gambiensis and G. morsitans submorsitans, in order to assess their population sizes (geographically and demographically), population densities and dispersal capacities. The two species exhibited markedly contrasting population structures. The first, G. p. gambiensis, displayed an isolation by distance with rather dense subpopulations exchanging immigrants at short distances (1.6 km/generation) and continuously from Sine-Saloum to Casamance, in line with the apparent continuity of the mangrove network found along this coast. In contrast, G. m. submorsitans presented a relatively dense population over a 75 km wide area, fragmented into sparsely distributed, very dense and interconnected pockets. These findings indicate that G. p. gambiensis likely constitutes a large metapopulation comprising 3266 effective individuals on average (and up to 7765), whereas G. m. submorsitans appeared as a single and large population, scattered in small pockets that could be readily replenished by immigrants from distant areas in case of control. These results suggest that elimination is not a realistic option for any of these two populations. Alternatively, localized integrated pest management is likely to be the most effective approach for controlling the negative effects of these biting flies on domestic animals, by reducing trypanosome transmission. However, further entomological investigations and the use of more informative genetic markers are needed to obtain a more precise understanding of the epidemiological processes involved.

Leishmania species are transmitted between mammals by sand flies, within which they undergo a complex developmental programme involving transitions between replicative and non-replicative life cycle forms. Recent work using single-cell transcriptomics has revealed the developmental pathways of Leishmania major in the sand fly, but it is unclear how faithfully such development is recapitulated during Leishmania culture in vitro. In addition, no work has revealed how gene expression changes across the cell cycle of replicative Leishmania forms. Using single-cell transcriptomics on unsynchronised cultures of L. major and L. mexicana, we addressed both questions. We show that in vitro L. mexicana promastigotes display the same two developmental endpoints seen in L. major cells recovered from sand flies, metacyclics and haptomonads. In contrast, L. major promastigote development in vitro does not predict the presence of leptomonads or haptomonads, although non-dividing late metacyclics form through an early metacyclic stage as parasites enter stationary phase. Using computational reconstruction of the cell cycle in procyclic promastigote forms of both Leishmania species identified hundreds of genes whose transcript levels vary as the cell cycle progresses, revealing core conserved processes between the two species and with the closely related Trypanosoma brucei. These genes allowed us to examine connections between life cycle and cell cycle progression, revealing that some developmental transitions are strongly linked to a cell cycle phase, whereas others occur asynchronously. This work provides a comprehensive atlas and marker gene list for the interlinked cell and life cycles of two laboratory-cultured Leishmania species.

Immune defenses may vary across environments due to differences in life history or abiotic and biotic characteristics, but few studies have compared the strength of immune responses across broad geographical gradients. Here we investigated how bacteria killing ability (BKA), a measure of constitutive innate immune function, varies among populations of tree swallows (Tachycineta bicolor) inhabiting different environments. We hypothesized that populations living in unpredictable environments, which also have shorter breeding seasons and thus fewer opportunities to renest in the event of reproductive failure, would invest in reproduction at the expense of immunity. To test this hypothesis, we measured BKA and monitored reproductive activity in four populations located across most of the breeding range of tree swallows, in Tennessee, New York, Wyoming and Alaska. Consistent with our predictions, BKA was lowest in Alaska and Wyoming and highest in Tennessee. In most populations BKA was also lower during incubation than during nestling provisioning, and lower in males than in females. These results suggest that the high reproductive investment seen in populations experiencing more unpredictable breeding conditions may come at a cost to immune function. While reduced immune investment could increase susceptibility to parasites, this potential cost is likely offset by lower parasite pressure in these environments, limiting the need for maintaining strong constitutive innate defenses like BKA.

Understanding the transmission and persistence of zoonoses in wild reservoirs is crucial for outbreak prevention, demographic assessment, and conservation planning. Host demography, ecology, behaviour and within-host viral kinetics together underpin transmission dynamics but are rarely united within epidemiological models. Here we characterised the transmission dynamics of European bat lyssavirus 2 (EBLV-2) infection dynamics by uniting a long-term study (2010-2018) of the ecology of five greater mouse-eared bat (Myotis myotis) colonies in France (~2700 marked individuals) with diagnostic evidence of virus circulation. Although no lyssavirus was detected in any of the 442 saliva samples tested, seropositivity showed synchronised peaks across colonies (0-100% annually), with the highest seroprevalence in 2015-2017, but remaining low otherwise. Fitting Bayesian compartmental epidemiological models, we found evidence that shifts in EBLV-2 prevalence were primarily driven by the gregarious behaviour of populations during the mating season and age-dependent variation in transmission. The duration of maternal immunity emerged as a key, and still empirically unknown factor, which shaped whether EBLV-2 maintenance was enzootic or required an external force of infection. Lyssavirus outbreaks in M. myotis were associated with few deaths overall and had a negligible impact on population dynamics. These findings underscore the value of integrating ecological and individual-level data with mechanistic modelling to enhance our understanding and management of zoonotic pathogens in wildlife.

Morphological variation in insects can indicate responses to habitat change and land use intensification, with potential consequences for trophic interactions. In insect host-parasitoid systems, host body is often a key determinant of offspring development and emergence success, as well as parasitoid host-selection behavior. Yet, most studies in agricultural landscapes have focused on size variation in crop-associated insects (i.e.: pests, pollinators and enemies), leaving those relying on non-crop resources largely unexplored. Here, we investigated how agroenvironmental contexts and trophic resource availability influence puparial size in Protocalliphora sialia (Diptera: Calliphoridae), an obligate hematophagous ectoparasite of Tree Swallow (Tachycineta bicolor) nestlings. We further tested whether puparial size affects emergence probability and risk of parasitoidism by Nasonia spp. (Hymenoptera: Pteromalidae) jewel wasps. Using 1,513 P. sialia puparia from 88 nests within a nest-box monitoring system spanning a 10,200-km^2 east-west intensification gradient in southern Quebec, Canada, we evaluated how agrointensive contexts, avian host availability, and ectoparasite densities influence puparial size, parasitoidism risk, and emergence success. The agricultural context appears to shape puparial size through survival biases and/or altered growth rates. In agroextensive contexts (i.e.: pastures, forages with high tree cover), where Protocalliphora are more numerous, P. sialia puparia were smaller and exhibited greater size variation. Conversely, in agrointensive environments (maize and soybeans monocultures with higher doses of pesticides), where Protocalliphora are fewer, small individuals were underrepresented, suggesting a selective mortality in P. sialia population. However, puparial size explained less than 10% of the variation in emergence or parasitism, likely reflecting consistently high parasitic pressure from Nasonia wasps, which infested 50-90% of puparia, depending on site and year. Our findings highlight the importance of selective mortality in structuring trait distributions and modulating multi-trophic interactions in human-modified landscapes.

Precocious reproduction, prior to the acquisition of the final adult morphology, occurs in a variety of animals and represents a distinct life-history strategy with important consequences for growth and survival. Larval reproduction is far rarer and usually restricted to groups that go through metamorphosis. In ctenophores, Carl Chun described dissogeny more than 130 years ago, a distinctive life-history pattern involving larval and adult reproductive periods separated by gonadal regression. Yet, the potential plasticity of larval reproduction in ctenophores remains unresolved. Here, we developed a method to follow and quantify reproductive output in Mnemiopsis leidyi larvae and tested environmental factors that may regulate precocious reproduction. We found that conspecific density strongly influences its onset: low-density, well-fed animals can bypass reproduction at the cydippid stage and develop rapidly into the lobed form, whereas higher density promotes precocious reproduction. Food availability and associated growth rate also contribute but cannot alone explain the observed differences. Once reproduction is established, however, high conspecific density suppresses reproductive output, an effect reproduced by exposure to seawater incubated with high M. leidyi densities, consistent with water-borne chemical communication. Salinity and light conditions further modulate spawning. Together, our results reveal substantial environmental plasticity in precocious reproduction in M. leidyi, reconcile long-standing contradictory observations of ctenophore development, and highlight how environmental cues can reshape reproductive life-history transitions.

Aedes albopictus (Skuse), the Asian tiger mosquito, is a highly competent arboviral vector whose range has expanded substantially across the United States over the past four decades. Despite predictive models placing Nebraska within the species' climatically suitable range, its establishment status in the state has remained poorly characterized. Here, we report results from a nine-year mosquito surveillance program (2017-2025) conducted across 44 Nebraska counties in collaboration with the Nebraska Department of Health and Human Services. Ae. albopictus was detected in five counties, with sustained, annually increasing populations documented in Richardson, Douglas, and Lancaster counties. Richardson County recorded continuous detections during 2017-2025, with proportional representation rising to 60.50% of collected mosquitoes by 2025. In Douglas and Lancaster counties, temporal advancement of first seasonal detection in 2024 and 2025 provide evidence consistent with successful overwintering rather than annual reintroduction. A cumulative degree-day model predicted adult emergence in mid-May across all county-year combinations, consistently preceding trap deployment by two to seven weeks and revealing a systematic early-season surveillance gap. Generalized linear mixed-effects models indicated that trap-level detection persistence, rather than urban location, was the primary predictor of yearly Ae. albopictus positivity, suggesting that current invasion dynamics are driven by focal source populations. These findings provide strong evidence for the establishment of Ae. albopictus in eastern Nebraska and highlight the need for earlier seasonal surveillance and standardized criteria to define establishment in northward-expanding vector populations.

Migration can increase population connectivity and gene flow but may also facilitate pathogen transmission among hosts and habitats. The way life-history strategies shape viral diversity in closely related freshwater fishes remains poorly understood. Here, we used metatranscriptomics to compare the viromes of two fish in the family Galaxiidae with contrasting life-history strategies: amphidromous [i]nanga (Galaxias maculatus) and river-resident Gollum galaxias (Galaxias gollumoides). We analysed 54 pooled libraries derived from 416 individuals collected across 27 sites spanning multiple catchments and fish-passage barriers in southern New Zealand. A total of 13 novel fish-associated viruses were identified from five viral families (Astroviridae, Circoviridae, Coronaviridae, Hepeviridae, and Picornaviridae). Virome composition differed markedly between species, with only a single virus-positive pool detected among [i]nanga, whereas Gollum galaxias harboured 11 novel viruses, with circoviruses dominating the virome. No fish-associated viruses were detected in post-larval juveniles of either species, and alpha diversity did not differ significantly among catchments, sites or positions relative to fish-passage barriers. However, beta diversity analyses revealed significant spatial structuring of viral communities among sites and catchments, indicating localised variation in virome composition. Contrary to our predictions, the migratory species exhibited substantially lower viral diversity than the river-resident species. The absence of detectable viruses in juveniles, combined with the greater viral richness of the longer-lived Gollum galaxias, suggests that host longevity and cumulative environmental exposure may be important drivers of virome assembly. These findings demonstrate that host life-history strategy influences virome composition and reveal unexpectedly low viral diversity in a globally widespread migratory galaxiid, providing new insights into virus-host ecology and connectivity in freshwater fishes.

The fieldwork was carried out between December 2023 and June 2025, with the aim, among others, of confirming the possible presence of the North African Gerbil (Gerbillus campestris) in Melilla. To this end, 56 camera trap stations were installed, each equipped with a camera trap. Of the total number of camera trap stations installed, the presence of Gerbillus campestris was detected in six of them, representing a capture success rate of 10.71%. Following the detection of the species, a specimen was captured using cage traps. For identification purposes, samples of epithelial tissue from the ear and faeces were collected. These samples were sent to the Institute of Vertebrate Biology of the Czech Academy of Sciences, Brno, Czech Republic, for analysis. There, DNA was extracted from the samples using the Innu PREP DNA Mini Kit 2.0 (IST Innuscreen GmbH, Germany) according to the manufacturer's protocol. The results obtained confirm, for the first time through records of the species, the presence of Gerbillus campestris in Melilla and, therefore, in Spain and Europe.

While mosquito invasions and associated pathogens are increasingly prominent in Europe, evidence gaps remain for many northerly regions where proactive management interventions may be most effective. At the regional periphery of Europe, there remains a significant knowledge gap concerning mosquito vector distributions in Northern Ireland. This study aimed to address this major gap by systematically surveying larval and adult mosquito populations in key habitats (ponds, reedbeds, saltmarsh, wet grasslands, wet woodlands, and urban gardens). We further identified environmental drivers of important vector species abundance and screened collected specimens with molecular techniques to identify presence of cryptic human-biting vector species. In Northern Ireland, mosquitoes (including potential bridge vectors of West Nile and Usutu viruses) were found to be widely distributed. Between May and August 2025, a total of 2430 mosquitoes (951 adults and 1479 larvae) were collected over 196 trap nights and 76 larval collections. Collections encompassed 17 species, with Culex pipiens/torrentium being the most widely distributed and abundant taxon recorded. Mosquito abundances were greatest in reedbed habitats, related negatively with rainfall and peaked in June and July. A subset of Culex pipiens/torrentium mosquitoes were identified to species/biotype level using PCR, with the majority (89.3%) identified as Culex pipiens pipiens and 9.3% as Culex torrentium. Importantly, the first detections of Culex pipiens/molestus hybrids (1.8%) on the island of Ireland were found. This work underscores the wide, yet previously undocumented abundance and distribution of potential zoonotic vectors in Northern Ireland and provides an important baseline for management planning and future research, while contributing to broader understandings of environmental population drivers and distributions.

Agricultural intensification is a major cause of population declines in farmland birds and insects worldwide. While these declines are expected to disrupt trophic interactions through density-dependent cascading effects, examples outside of crop pest control or involving more than two trophic levels remain poorly documented. Here we quantify such cascading effects across three trophic levels involving a declining aerial insectivore, the Tree Swallow (Tachycineta bicolor), which breeding success decreases with agricultural intensity, Protocalliphora bird blowflies (Diptera: Calliphoridae) that parasitize its nestlings, and Nasonia parasitoid jewel wasps (Hymenoptera: Pteromalidae) that feed upon blowfly pupae within swallow nests. Agriculture intensity may affect each of these actors either directly via, for instance, toxicological effects from pesticide use, or indirectly through density-dependent trophic interactions, which can exacerbate or mitigate one another. Using Bayesian models within a causal inference framework, we quantified how nestling host density, meteorological conditions, and agricultural landscape context affect Nasonia parasitoid infestation of Protocalliphora pupae, and estimated Protocalliphora emergence success. Analyses were based on 3,340 Protocalliphora puparia collected from 248 Tree Swallow nests over a 16-year period (2004-2019) across a 10,200-km2 agricultural intensity gradient in Southern Quebec, Canada. Model predictions indicated that Protocalliphora emergence probability increased steeply with nestling host availability (i.e., nestling-days). Whereas Nasonia parasitoid infestation occurrence at the nest level increased with Protocalliphora host availability, the proportion of infested Protocalliphora pupae within nest decreased and led to a dilution effect at the individual level. Although Protocalliphora abundance within nests decreased with agricultural intensity, their emergence probability peaked in agrointensive landscapes. This increase was associated with reduced Nasonia infestation rates, suggesting that agricultural intensity indirectly mitigated parasitoid pressure on Protocalliphora host populations. Our results highlight how agricultural intensification can shape multitrophic interactions and life-history traits (emergence success and infestation rates) by simultaneously reducing host availability and weakening top-down control by parasitoids.

In Baltic grey seals (Halichoerus grypus), interannual variation in sea-ice availability can alter the proportion of females breeding on land, generating substantial variation in local breeding density among years. We investigated preweaning survival, post-weaning residence, and weaning body condition at a Baltic grey seal land-breeding colony over two years with contrasting colony densities. Drone imagery provided repeated counts of pups in distinct developmental stages and estimates of moulting pup mass as an index of weaning body condition. A Bayesian state-space population model estimated substantially greater pup production in the high- (282.5 [95% CrI = 252.8 to 320.9] pups) than the low-density (188.3 [95% CrI =168.9 to 210.8] pups) year, but similar preweaning survival (0.66 [95% CrI = 0.53 to 0.81] and 0.67 [95% CrI = 0.56 to 0.79], respectively). In contrast, post-weaning residence differed substantially between the high- (11.8 [95% CrI = 9.2 to 15.7] days) and low-density (30.3 [95% CrI = 23.1 to 39.1] days) years. Greater exposure to other pups during early life was associated with lower weaning body mass ({beta} = -0.45 [-0.89 to -0.01], P[{beta} < 0] = 0.98). Together, these results suggest that the consequences of higher breeding density may emerge through poorer weaning condition and altered post-weaning residence even when effects on preweaning survival are not detectable.

When mating among genetically related individuals results in inbreeding depression, various behaviors and life-history traits evolve to favor outbreeding. Long-range sexual communication following natal dispersal is one such adaptation. In parasitoid wasps with single-locus complementary sex determination (sl-CSD), inbreeding increases the production of unviable or sterile diploid males. Mastrus ridens is such a species, with a genetic load underpinned by sl-CSD, combined with a high potential for sibmating because siblings emerge in close proximity from the same host and become sexually active shortly after emergence. With this context in mind, we investigated the mate-finding strategy of M. ridens using complementary behavioral, chemical, and electrophysiological approaches. Wind-tunnel bioassays and field experiments showed that males orient toward virgin females and, to a lesser extent, mated females. This attraction from a distance appeared to be the sole form of conspecific attraction. Chemical analyses using static and dynamic headspace sampling and solvent extracts, followed by GC-MS, revealed several potential pheromonal candidates, including methyl 6-methylsalicylate. This compound elicited reproducible antennal responses in males in GC-EAD recordings. Altogether, our results suggest that mate finding in M. ridens is guided by a volatile sex pheromone emitted by females, with methyl 6-methylsalicylate as one putative component. Such mate-finding mechanisms promote outbreeding and may explain the low proportion of unfit diploid males observed in natural populations. Surprisingly, chemical analyses suggested that males also emit this compound, whereas neither males nor females showed behavioral responses to males. This apparent discrepancy suggests that attraction may depend on a more complex pheromonal signal and opens further avenues for investigation.

Sporozoites, the infectious stage of Plasmodium malaria parasites, must invade two different tissues in two very different hosts to ensure transmission: the mosquito salivary glands and the human liver. Whether infectivity to hepatocytes is affected by physiological components, such as the availability of nutrients within the Anopheles female, is not well understood. Here we show that Plasmodium falciparum sporozoite infectivity to hepatocytes is increased if mosquitoes take an additional blood meal while oocysts are developing. Infection rates of HC04.J7 hepatocytes at 24 h were 1.8-fold higher for P. falciparum salivary gland sporozoites isolated from Anopheles gambiae females fed twice (2BF) compared to those which only received the infectious blood meal (1BF). This increase in infectivity was observed at an early infection timepoint (day 11) and was accompanied by a marked shift in the transcriptional profiles of 2BF salivary gland sporozoites, identifying genes likely involved in infection and development in the mammalian host. Overall, our data reveal new factors potentially involved in sporozoite infectivity, and have broad implications for understanding transmission of malaria in endemic areas where multiple blood meals commonly shape available resources for adult female mosquitoes.