Peer Community Journal

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.

Parasites decrease host fitness, more so during physiologically challenging life stages such as migration. However, parasites are unevenly distributed among individuals, and hosts with different physiological condition may face different costs of migration with infections. We tested whether the relationship between infection by haemosporidian parasites and oxidative balance during autumn migration in blackcaps depends on individual age and sex, two major sources of variation in prevalence and physiological condition. We captured blackcaps at stopover in central Spain and measured infection status and physiological parameters. Blackcaps physiologically more prepared to depart from stopover had lower antioxidant capacity of plasma, probably because of physiological adjustments prior to the upcoming flight stage. Plasmodium parasites occurred only in coinfections, which were more frequent among young blackcaps. Regardless of individual sex and age, coinfected birds had lower antioxidant capacity of plasma, and blackcaps coinfected with Plasmodium had more oxidative damage to lipids in their red blood cells. Our results support the idea that haemosporidian infections impair the oxidative balance of blackcaps during autumn migration, which could impair individual performance and ultimately fitness, with increased population impacts during the first year of life when a larger proportion of individuals migrate coinfected with harmful Plasmodium parasites.

Lateral gene transfer (LGT) from organellar to eukaryotic genomes is ubiquitous, resulting in the presence of widespread nuclear mitochondrial DNA segments (NUMTs) and/or plastid gene transfers. While LGT from bacterial associates of eukaryotes is generally less common, many arthropod genomes are littered with LGT originating from Wolbachia (order Rickettsiales) - a vertically transmitted, obligate intracellular symbiont of invertebrates. This contrasts with Rickettsia, a related genus including many important human pathogens, which has not been shown to promulgate LGT within its arthropod vectors. Here, we present the 8.6 Gb genome of a cell line derived from A. variegatum (the tropical bont tick), which is the primary vector of Rickettsia africae (agent of African tick-bite fever). In addition to numerous NUMTs and diverse families of transposable elements, an almost-complete chromosome of R. africae origin was identified in the cell line genome, localised to the putative sex chromosome. Sequencing of field-collected A. variegatum confirmed the presence of this large LGT in wild ticks, although the absence of transferred genes from the R. africae plasmid provided a means to differentiate LGT from genuine rickettsial infections. These findings highlight the imperative to consider the possibility of LGT when screening vectors for pathogens by PCR.

1. Species monitoring programmes regularly encounter missing data, which complicates tasks such as estimating population size or detecting temporal trends. Selecting an imputation method suited to the properties of the data is therefore an important practical challenge, particularly for species exhibiting overdispersion and zero-inflation. 2. We compare seventeen imputation methods, comprising thirteen Poisson-based statistical models (accounting for overdispersion and zero-inflation, with fixed, random and multivariate structures) and four contrast-based approaches (LORI, MICE, missForest, correspondence analysis). Using four complete monitoring datasets of waterbird species surveyed across France and Italy over 21 years, illustrating a variety of abundance distributions, we introduced missing observations under a realistic Missing At Random mechanism, at rates from 5% to 70%. We evaluated all methods on computational burden, prediction accuracy for positive and null counts, quantification of uncertainty, and accuracy of population-size estimates. 3. No single method dominates across all criteria. Statistical models and contrast-based approaches yield similar point predictions, but only statistical models provide a genuine measure of uncertainty. Models that jointly account for overdispersion and zero-inflation perform best at predicting zero counts and achieving reliable prediction intervals, and the most suitable method ultimately depends on the abundance distribution of the target species. 4. These results provide practical guidance for ecologists selecting an imputation strategy for incomplete count data, highlighting the trade-offs between predictive accuracy, computational cost, and the ability to propagate uncertainty through subsequent ecological analyses such as population-trend estimation.

Selection-intensity estimates from RELAX are reported as a point value of K with a likelihood-ratio P. We report that, at default settings and on data of ordinary size, the program does not reproduce its own fits. Of 27 enzyme entries refitted under two optimiser configurations, none reproduced its log-likelihood to within 0.01 units; the median change was 103 units, the largest over 3,400, and four verdicts reversed. Eighty null orthologues reproduced none. A byte-identical command returned a distinct likelihood on every repetition, single-threaded, across three releases, and on alignments simulated under the fitted model, where 3.3 per cent of replicates reproduced. The documented random-number seed never reaches the generator when assigned on the command line, yet reads back as the value supplied. PAML localises the cause: its two-ratio model, without site classes, reproduced its log-likelihood for all 288 genes; its site-class models agreed for 27 to 67 per cent. The instability follows the mixture over sites, not the program. The output does not show it: 46 of 410 fits ended with a negative likelihood-ratio statistic, impossible under convergence, and 123 of 410 report a K re-estimated under a domain restriction rather than the unconstrained maximum. Of 234 published studies using RELAX, none reported a seed. Seeding while holding the thread count at one reproduced sixty of sixty runs on twenty genes under two releases; the seed alone reproduced none of five, and no documentation states the second condition. We recommend that fits be repeated and their dispersion published.

The goat research community benefits from one of the most comprehensive livestock genome assemblies, ARS1. However, the rich diversity of goat breeds suggests that a single individual genome is not enough to represent the species full genomic content. Third-generation sequencing technologies have recently made it possible to generate very high quality assemblies that should enable to elucidate unique and/or complex phenotype determinisms depending on genetic structural variations. In the SeqOccin project, nine goat breeds were selected to represent French goat genetic diversity: Alpine, Boer, Creole, Fosses, Poitevine, Provencale, Pyreneenne, Rove and Saanen. High-molecular-weight DNA was extracted from blood samples and subjected to long read whole-genome sequencing. There reads were used to generate de novo assemblies and were aligned onto the public reference to call structural variants. PacBio HiFi and parental Illumina reads were used to assemble phased haplotypes of the Alpine breed. For the other breeds, PacBio raw (CLR) or ONT reads were generated. Our work demonstrates the feasibility of assembling genomes with a reasonable amount of data and appropriate software packages, delivering one high-quality assembly and 8 standard goat assemblies that pave the way for pangenome construction and structural variant functional studies.

Recent work on the design of spatial capture-recapture (SCR) studies has largely focused on settings with a single set of movement and detection parameters. In many applied contexts, however, populations are naturally stratified-for example, when different sexes exhibit markedly different movement scales and encounter processes, or when more than one focal species is monitored. Designs that perform well for one group may be inefficient or suboptimal for others, despite the substantial logistical effort typically required to implement an SCR survey. We describe several potential SCR design strategies for surveying multiple groups, and compare these using simulation. Designs considered include regular and optimised grids, clustered and lacework designs, and designs generated using genetic algorithms, including new approaches that optimise information from both groups and a recently published two-stage design. Using proximity detectors such as camera traps, we evaluate design performance in terms of precision, bias, and coverage, and examine how performance changes with the number of available resources. Our results show that multi-group SCR surveys can perform well even when focal groups differ greatly in movement scale and encounter rate, provided that some simple guidelines are followed. Systematic layouts accommodating more than one characteristic spacing e.g. cluster designs, performed consistently well and are a reliable default option. Regular grid designs with a single spacing between detectors are feasible when differences between groups are moderate but performed poorly as differences increased. Establishing when these designs become problematic adds unnecessary risk to the design process. Algorithmic designs did not outperform systematic designs even in highly-skewed scenarios expected to favour them. In practice, when there is more than one focal group we recommend identifying the smallest and largest plausible movement scales and rejecting potentially pathological designs, primarily those that cannot generate recaptures at the smaller scale or cover a substantial area at the larger scale.

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.

Baker's rule predicts that selfing is a beneficial trait for colonizers, whose prediction in the kingdom Fungi is reinforced by recent work showing the invasive ectomycorrhizal basidiomycete Amanita phalloides with self-incompatibility (SI) system has evolved unisexual reproduction in its introduced range. Unlike in plants, the multiallelic SI system of basidiomycetes does not fully prevent selfing among the meiotic offspring of a single dikaryotic (diploid) parent. This distinction allows observation of an intermediate state of selection where selfing is preferred in a species with SI system during colonization before a change in self-incompatibility architecture. Here, we analyzed the genomes of 189 isolates of the ectomycorrhizal basidiomycete Suillus luteus collected from both its native European range and five independently introduced regions to test whether selfing rates increase and whether the SI system had degraded after introduction. Unlike the A. phalloides case, we found no evidence of unisexual reproduction or loss of the SI system in the introduced ranges of S. luteus. Rather, all introduced populations maintained outcrossing rates higher than random mating as in the native populations. However, selfing events were detected in three individuals from a single introduced population, representing the first documented case of selfed sporocarps of Boletales fungi. Our works in S. luteus demonstrates that although selfing is possible in this species, it is not favored during colonization, potentially due to inbreeding depression and higher population density, highlighting the complex interaction among the invasion biology, genetics, and the colonizing context.

1. Plant roots employ multiple strategies to acquire soil resources, including the outgrowth of root hairs and the association with arbuscular mycorrhizal fungi (AMF). These structures represent alternative pathways to access soil resources. Evidence is derived mostly from studies within single species or multi-species experiments under controlled conditions. It remains unclear whether this trade-off holds under natural conditions and whether it manifests consistently across different scales of biological organization. 2. We analysed 81 grassland species representing grasses, forbs and legumes, sampled under heterogeneous field conditions in three regions across Germany, allowing for analyses (i) across different species (interspecific), (ii) across communities (intercommunity), (iii) within plant species (intraspecific) and (iv) within root systems of individual plants (intra-individual). Fine root traits related to AMF colonization (including scoring of fungal hyphae and arbuscules) and root hair growth (including length and incidence) were quantified. 3. Comparing plant species, we observed a strong negative correlation between mycorrhizal colonization and root hair traits, confirming previous studies working under controlled conditions. The negative relationship between root hair length and mycorrhization was also evident when comparing mean trait values at the plant community scale. Among different individuals of the same species (i.e. intraspecific scale), there was no consistent pattern of trait-trait relationship. When comparing within roots of the same plant (i.e. intra-individual scale), co-occurrence of root hairs and mycorrhiza was less likely than what would be expected by chance, with forbs showing the most consistent negative pattern. 4. Our study demonstrates that the hypothesized trade-off between AMF and root hairs is a robust pattern under field conditions at the community and interspecific scale. At the intraspecific scale, no uniform trade-off pattern can be discerned, and within single root systems the negative trade-off pattern is widespread but weak. This indicates that ecological and evolutionary constraints shape trait-trait relationships at higher scale, whereas physiological limits do not restrict trait co-occurrence at lower scales.

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.

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.

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.

The sheep research community benefits from a genome assembly that has been refined over the last 15 years. However, the rich diversity of sheep breeds suggests that a single individual genome is not enough to represent the species full genomic content. Third-generation sequencing technologies have recently made it possible to generate high quality assemblies that should enable to elucidate unique and/or complex phenotype determinisms depending on genetic structural variations. In the SeqOccin project, twelve breeds were selected to represent (mostly French) sheep genetic diversity: Basco-Bearnaise, Black Belly, Blanche du Massif Central, Corse, Ile de France, Lacaune, Manech Tete Noire, Noire du Velay, Ouessant, Rambouillet, Vendeen, Soay. High-molecular-weight DNA was extracted from blood samples and subjected to long read whole-genome sequencing. The reads were used to generate de novo assemblies and were aligned onto the public reference to call structural variants. PacBio HiFi and parental Illumina reads were used to assemble phased haplotypes of the Lacaune breed. For the other breeds, PacBio raw (CLR) or ONT reads were generated. Our work demonstrates the feasibility of assembling genomes with a reasonable amount of data and appropriate software packages, delivering one high-quality assembly and 11 standard sheep assemblies that pave the way for pangenome construction and structural variant functional studies.

Highly pathogenic avian influenza (HPAI) is among the most significant disease threats to poultry worldwide. Modelling challenges, where independent teams address a common epidemiological scenario, can support the development of response capabilities and strengthen communication between modellers and risk managers. Despite the established benefits of these exercises, they remain uncommon in animal health, and none have previously addressed avian influenza. Here Jolly Island, an artificial landscape of 9160 poultry farms with five production types, was developed and seeded with stochastic HPAI epidemics that ran their course over a three-to-four month period. From January through April 2026, 45 registered teams across five continents investigated a selected synthetic epidemic, producing probabilistic forecasts, inferring transmission drivers, and evaluating counterfactual control policies. Here the objectives, design choices, and data generation of the HPAI Modelling Challenge are described in full, covering transmission processes, surveillance system operations, and control strategy implementation, along with the selected epidemic trajectory, the narrative situation reports supplied to the teams at each phase, and the complete phase-specific datasets released to them. As the opening paper of this special issue, here is provided the foundation upon which all subsequent contributions, where teams document their approaches and analyses, are built.

BACKGROUND: The Argentine ant (Linepithema humile) is among the world's worst invasive species. In the Donana Biological Reserve (Spain), it is associated with human structures that provide the mild, moist conditions it needs within the hot, dry scrubland. We evaluated a habitat-modification treatment, predicting that removing this suitable habitat would eradicate the invader locally. Over an 18-year period, we monitored ant communities at a treated and an untreated site, at different distances from the human structures, before and after treatment, and analysed how temperature and worker numbers affected encounters at baits between the Argentine ant and native ants. RESULTS: Distance to the human structures shaped ant communities at both sites, but only the treated site changed after treatment. There, the Argentine ant decreased close to the human structure and almost disappeared farther from it, whereas the endemic Cataglyphis tartessica and C. floricola recovered, especially close to the structure. At baits, the outcome of encounters depended mainly on how temperature changes favoured each species relative to its maximum activity temperature: Cataglyphis spp. displaced the Argentine ant in the morning, as temperature increased, and the Argentine ant recovered the baits in the late afternoon. Numerical advantage favoured only the Argentine ant. CONCLUSION: Habitat modification reduced the Argentine ant and allowed endemic ants to recover, but did not eradicate the invader, which persisted at low abundance 13 years after treatment. In protected areas, habitat management should be combined with eradication, monitoring and prevention of spread.

Coinfections are common in a nature and often involve competition among coinfecting parasites to extract resources from the host. As a consequence of this competition, parasites can face trade-offs between investing in competition with other parasites and exploiting the host for their own growth and transmission. Such within-host interactions among parasites can either increase or decrease virulence depending on how they alter parasite density, resource use, and host damage. We tested the inhibitory effect of different species of the entomopathogenic fungus Metarhizium on one another in vitro and the impact of coinfection on virulence towards Tribolium castaneum. We found that Metarhizium pemphigi is able to inhibit all other isolates of Metarhizium tested, whereas Metarhizium pingshaense was relatively resistant to inhibition by other Metarhizium strains. Virulence towards the host, Tribolium castaneum, differed very little among isolates tested. We did, however, observe differences between virulence in single and coinfections in two cases. Where there were virulence differences, these were both consistent with in vitro inhibition patterns and interestingly resulted in reduced virulence in coinfection. This is inconsistent with broader patterns of virulence in coinfection, where virulence seems to be driven by the more virulent parasite. It is, however, consistent with data from other systems where there is evidence of interference competition. Our data thus support the idea that, in contrast to other forms of between parasite interaction, interference competition results in reduced virulence in coinfection.

A complete reference genome for species with chromosomally-determined separate sexes should contain scaffolds for all sex chromosome homologs. However, sex chromosomes present distinct computational challenges compared to autosomes. Here we present a k-mer based analysis that utilizes whole-genome sequencing of a few sex-identified isolates: Cytogenetics-By-Sequencing (CBS) tools. Unlike other approaches that typically address one aspect of the sex chromosomes, CBS-tools strives to guide users from the discovery of the heterogametic sex through identifying the sex-determination region (SDR). The core of CBS-tools is automated quantification of sex-specific k-mers in order to predict the heterogametic sex. Using publicly-available datasets, CBS-tools correctly identified the known sex-system of the 31 species tested. Additionally, we used these k-mers to verify and correct phasing of sex chromosomes between haplotypes in species representing different sex-systems. Finally, we used these k-mers to delimit the SDR boundary using an interactive web platform. CBS-tools was developed with previously unexplored sex chromosome systems in mind, but is also suitable for well-examined sex chromosome pairs.

Sperm competition can occur when sperm from two or more males compete for fertilization of a batch of eggs. While it has been extensively studied in internal fertilizers, it remains poorly under-stood in external fertilizers. To investigate how sperm competition affects fertilization under sperm limitation, we conducted controlled experiments on two sea urchin species: Paracentrotus lividus and Dendraster excentricus. Eggs from a single female were exposed to sperm from three individ-ual males separately (non-competitive treatment) or to a pooled sample from three males (competi-tive treatment). We measured fertilization success as proportion of eggs fertilized after two hours and assessed sperm motility as percentage of motile sperm and curvilinear velocity. For P. lividus , we estimated the sperm concentrations where fertilization success was 50% of maximum observed fertilization success (F50), and the fertilization efficiency of the sperm (Fe), i.e. the likelihood that a sperm - egg encounter results in a fertilizing contact, using the fertilization kinetics model of Styan (1998). For D. excentricus, fertilization success was measured at a sperm concentration expected to produce 50% fertilization success. Sperm competition reduced fertilization success and increased F50 compared to non-competitive treatments, suggesting antagonistic effects that may compromise fertilization under sperm-limited conditions in external fertilizers.

The Peruvian Andes, and especially its slopes towards the Amazonian Basin are well-known biodiversity hotspots for plants and animals, which represent the visible part of biodiversity. However, not much is known about microeukaryotic diversity, despite the significant role protists play in the ecosystem. Based on a molecular survey of the diversity of Arcellinida (a group of amoebae well known for its environmental sensitivity and restricted geographical distributions), we estimated its diversity along an elevation gradient spanning more than 3000 meters. Surprisingly, Neotropical forest Arcellinida were less abundant and diverse than in most other extratropical biomes; nevertheless, communities differed strongly in their taxonomic composition as compared with forests in higher latitudes. Notably, sequences related to Trigonopyxis were most abundant, whereas this group occurs only marginally in extratropical settings. Elevation influenced beta diversity, showing different assembly processes occurring at different elevation belts. Canopy cover positively influenced both phylogenetic and taxonomic diversity, showing the importance of above and below ground integration in tropical forests.