Small Vertebrates Research Articles

Dolomedes fishing spider biology: gaps and opportunities for future research

Dolomedes may easily be considered to be among the most charismatic spider taxa. Known colloquially as fishing or raft spiders, this clade of dolomedid cursorial hunters is speciose with about 100 valid species names. Most Dolomedes are large spiders that inhabit water bodies across all continents except Antarctica and, interestingly, South America. Dolomedes have captured the attention of researchers and the public alike for their ability to walk on and submerge under water, fish for prey (including small vertebrates), and for their often-bizarre mating behavior that includes examples of male spontaneous death and sexual cannibalism. In this review, we critically evaluate what is known of Dolomedes biology, focusing on their systematics and morphology, ecology, behavior, and conservation. Given their close association with water, Dolomedes may be particularly vulnerable to the impacts of anthropogenic change and provide an important group of indicator species for understanding the effect of pollution, habitat loss and climate change. We outline a roadmap for future studies that, in our view, will consolidate Dolomedes as an ideal model lineage among spiders for addressing a vast array of questions across multiple fields of biology.

Early Alarm on the First Occurrence of the Southern Giant Hornet Vespa soror du Buysson, 1905 (Vespidae) in Europe.

An eco-monitoring program to assess the biodiversity of insects affected by yellow-legged hornet (Vespa velutina) trapping in the north of the Iberian Peninsula (Spain) revealed the first occurrence of the southern giant hornet Vespa soror (Hymenoptera, Vespidae) on the European continent. We present a detailed characterization, combining morphological characteristics and molecular tools for genetic identification, as well as key information on its identification with respect to other hornets found on the Iberian Peninsula. We discuss the most plausible pathways and vectors of introduction, its potential invasiveness, and subsequent impacts on host localities. Our preliminary results raise concerns about the potential threat of V. soror to human health and ecosystem dynamics, as it is a highly predatory species on other insects and even small vertebrates. Finally, this study confirms once again the usefulness of studying insects trapped in such traps for rapid response and early detection of inland invasive species. We also propose a common Spanish name for the species, "avispón sóror".

Habitat structure and an introduced predator limit the abundance of an endangered ground-nesting bird.

Understanding the factors that limit the abundance of threatened species is critical for the development of effective conservation strategies. However, gaining such knowledge from monitoring programs and using it to inform decision-making for rare species can be difficult due to methodological issues posed by the problems of distinguishing true absences from false absences and the analysis of datasets dominated by zero counts. The plains-wanderer (Pedionomus torquatus) is a critically endangered ground-nesting bird that occurs in grasslands of southeastern Australia. Decline of the plains-wanderer has been attributed to habitat modification but little emphasis has been placed on the role of introduced predators, such as the red fox (Vulpes vulpes), which have had a devastating effect on small ground-dwelling vertebrates in dryland regions of Australia. Here, we use a 9-year time series of spotlight counts to investigate the impact of vegetation structure and fox presence on plains-wanderer occupancy and abundance. We used distance sampling to determine the effective strip width for sighting plains-wanderers during spotlight surveys. We then used a hurdle model approach whereby binomial generalized additive models were fitted to presence/absence data within the effective strip-width across all sites and negative-binomial models were fitted to an index of abundance at sites where plains-wanderers were observed. Plains-wanderer occupancy and abundance fluctuated markedly through time. Where foxes were absent, occupancy (but not abundance) of plains-wanderers showed a humped relationship with grass height with an optimal height between 50 and 150 mm. Where foxes were present however, this relationship broke down and plains-wanderers were rarely recorded. Our results suggest that plains-wanderers should benefit from management strategies that maintain grass height at optimal levels and exclude foxes or effectively suppress their populations. A key message from this study is that if statistical analyses of data generated by monitoring programs for rare species are intended to inform management decisions by identifying relationships between threatened species and drivers of their abundance, there should be consideration of analytic approaches that account for true and false zeroes, high prevalence of zeroes, and the possibility of nonlinear responses.

The initial impact of a predator exclosure sanctuary on small vertebrates in semi‐arid Western Australia

AbstractPredator exclosure reserves have become pivotal to conserving some threatened Australian mammalian fauna. However, the impact of predator exclosure reserves on resident (non‐target) small vertebrates is less understood. Given the potential ecological consequences of fencing, and the contribution of small vertebrates to ecosystem processes, we investigated species' occupancy, and site colonization and extinction dynamics, inside and outside of a predator exclosure fenced area for nine small vertebrates (six reptile, three mammal). We conducted vertebrate surveying 4 years (2011–2014) pre‐ and 2 years (2015–2016) post‐fence construction at the Australian Wildlife Conservancy Mt Gibson Sanctuary, in the semi‐arid, critically endangered Wheatbelt Eucalypt Woodlands of Western Australia, to assess spatial and temporal trends in small mammal and reptile populations. Of nine species modelled, one reptile (Diplodactylus pulcher) decreased in occupancy inside of the fence following construction. We detected no negative impact of the predator exclosure fence on the remaining eight species. One native reptile species (Lerista kingi) and an introduced rodent (Mus musculus) increased occupancy inside (but not outside) the fenced area. Spatial and temporal occupancy inside and outside of the fenced area for all other modelled species was stable. Biotic (habitat) factors and stochastic interannual detectability were the most significant drivers of species' occupancy. Species‐specific habitat associations were partially determined by site‐based presence and absence probabilities. This suggests that the predator exclosure sanctuary either benefitted or had minimal impact on all but one modelled species. These results may be a useful reference for future fencing projects aimed at both conservation and land management.

Carcass size, not source or taxon, dictates breeding performance and carcass use in a burying beetle.

Small vertebrate carcasses represent critical resources for many terrestrial organisms, including burying beetles, which rely on carcasses for survival and breeding. Carcass attributes can influence the reproduction of burying beetles, yet most studies on their breeding ecology have used laboratory-reared carcasses of limited sizes. We conducted breeding and feeding experiments using a wide size range of lab (laboratory mice) and wild carcasses (wild mammals, birds and reptiles) to investigate how carcass size, source and taxon affect various breeding outcomes (e.g. clutch size, brood size and brood mass) of the burying beetle Nicrophorus nepalensis. Our results reveal a hump-shaped relationship between carcass size and breeding performance, with optimal breeding outcomes occurring on medium-sized carcasses. Furthermore, despite the variation in carcass tissue nutritional composition, breeding outcomes and larval growth did not differ between the two carcass sources or among the three wild carcass taxa. Finally, we found a larval quality-quantity trade-off across the range of carcasses examined, with carcass size shaping the larval life-history traits. Overall, these results elucidate how carcass resources may influence the breeding performance of burying beetles. Importantly, our study provides solid evidence validating decades of research using lab carcasses to study the reproductive ecology of burying beetles.

Long-term anthropogenic drivers of diet composition and niche breadth in an endangered avian scavenger: The scant role of sanitary regulations

The widespread and accelerated transformation experienced by our planet results in populations of endangered species having to survive in highly-altered environments and increasingly relying on resources provided by human subsidies. Presently, vultures rank among the most imperiled functional groups of large vertebrates globally, a situation that is largely attributed to the rapid intensification of agriculture and the subsequent transformation of farming practices and agro-grazing landscapes. Hence, documenting the long-term fluctuations in the vultures’ dietary patterns in response to environmental changes, particularly in livestock management, is of utmost significance. Here, by means of bulk stable isotope analysis (δ13C and δ15N), we examined long-term variations in diet composition and niche breadth in a population of the endangered Egyptian vulture (Neophron percnopterus) in the north of the Iberian Peninsula. We specifically focused on the effect of European sanitary regulations enacted in 2014, which allow sheep farmers to abandon livestock remains for consumption by scavenging birds, except for pig carcasses. We found that the implementation of these new regulations did not cause a change in the diet of vultures. Changes were only detected in scrubland areas and were driven by an increase in the consumption of wild rabbit which, in turn, lead to expansion of the trophic niche. These results indicate that these endangered vultures are heavily dependent on food sources derived from intensive farming, mostly of pigs illegally disposed in the wild, and moreover, highlight the importance of carcasses of small vertebrates, often disregarded in conservation programs in favor of the contribution of livestock remains. Our research provides valuable insights into the spatiotemporal dynamics of trophic strategies among avian scavengers, specifically in the context of changes resulting from intensified human-pressures on agro-grazing Mediterranean landscapes.

THE VERTEBRATE FAUNA OF THE LOWER PLIOCENE CONTINENTAL DEPOSITS IN THE EBRO DELTA (BAIX EBRE BASIN, NE SPAIN)

The Pliocene faunas of the northeastern Iberian Peninsula are poorly known due to the scarcity of deposits from this interval. In this context, the site of Sant Nofre-Campredó (Baix Ebre, Catalonia, Spain), comprising two contemporaneous outcrops belonging to geologically correlated sections (Sant Nofre and Campredó-Via Fèrria), makes a significant contribution to the study of Pliocene faunas in this region. The first vertebrate remains were recovered in the late 1970s in clay deposits close to the Sant Nofre Hill. Later, intensive sampling in the 1980s at the Campredó-Via Fèrria outcrop led to the recovery of more than a hundred fossil specimens, including small vertebrates and large mammals. Here, we revise these collections to expand and update the vertebrate list, better constrain the age of the continental deposits, and characterize the environment and climate around the mouth of the Ebro River during the formation of the site. Up to 24 vertebrate species have been identified at Sant Nofre-Campredó, including amphibians, reptiles, and mammals. The presence of the rodents Paraethomys baeticus Piñero & Verzi, 2020, Apocricetus barrierei (Mein & Michaux 1970), and Occitanomys brailloni Michaux, 1969 implies an early Ruscinian age (MN14 unit) for the two outcrops. Biostratigraphic correlations with other well-dated sites in the Iberian Peninsula refine the estimated age of Sant Nofre-Campredó to approximately 4.6 Ma. The ecological affinities of the identified fauna reveal a landscape dominated by woodland habitats in the surroundings of the site during the Early Pliocene, with presence of open humid meadows and water bodies under warm and moist climatic conditions.

DnaA modulates the gene expression and morphology of the Lyme disease spirochete.

Lyme disease is the most prevalent tick-borne infection in the Northern Hemisphere. Borrelia burgdorferi , the causative spirochete bacteria, has been maintained in nature for millennia in a consistent enzootic cycle between Ixodes ticks and various small vertebrate hosts. During the tick's blood meal, B. burgdorferi substantially increases its replication rate, alters its repertoire of outer surface proteins, and disseminates into the new vertebrate host. Across eubacteria, DnaA is the master regulatory protein that initiates chromosomal replication and acts as a transcription factor to regulate specific pathways. Here, we describe the roles that B. burgdorferi DnaA has on the physiology and gene expression of this medically important pathogen.

Verifying the Diet of the Slender Racer, <i>Orientocoluber spinalis</i> (Peters, 1866), Rare, Threatened, and Endangered Across Northeast Asian Countries

Understanding the diet of a species provides important information on various aspects of its ecology. Diet studies are few on the reptile species, of which observation frequency is very low, due to the great difficulty of sampling. We investigated the diet of Orientocoluber spinalis, a long and slender small grassland snake with high rarity and a high extinction risk across northeast Asian countries, by applying regurgitated stomach content analysis and metabarcoding analysis using a developed O. spinalis blocking primer. Among 19 diet samples, which were mainly collected on islands along the west coast of the Korean Peninsula, we determined the food sources of 13 samples, which consisted of four small vertebrate species, the Tsushima smooth skink (Scincella vandenburghi), mountain grass lizard (Takydromus wolteri), Japanese keelback (Hebius vibakari), and lesser white-toothed shrew (Crocidura shantungensis). Among them, O. spinalis most frequently fed on S. vandenburghi (10 of 13 cases). Our results show that O. spinalis mainly forages small reptiles and mammals in grasslands adjoining to lowland forests and could be beneficial for conservation in the field and rearing of this vulnerable species in the facilities.

Fire protection priorities in the oak forests of Iran with an emphasis on vertebrate habitat preservation

This study examines the impact of fire incidents on wildlife and habitats in the western oak forests of Iran (Zagros region). These forests are globally recognized for their exceptional biodiversity but are frequently threatened by wildfires. To achieve this, the study uses the space–time scan statistics permutation (STSSP) model to identify areas with a higher frequency of fires. The study also analyzes the effects of fires on the Zagros forests from 2000 to 2021 using remote-sensing MODIS data. Also, to understand the elements at risk of fire, burned areas were assessed based on the richness of vertebrate species, determined by the distribution of 88 vertebrate species. The results show that the annual fire rate in the Zagros forests is 76.2 (fire occurrences per year), calculated using the Poisson distribution. Findings show the highest fire rates are found in the northwest and a part of the south of the Zagros. The northwest of the Zagros also has the largest number of single fires and clusters, indicating a wide spatial distribution of fire in these regions. On the other side, it was unexpectedly found that these regions have the richest number of species and higher habitat value. The results demonstrate a significant correlation between the value of the habitat and the extent of burned areas (p < 0.05). The study also reveals that the greatest impact of fires is on small vertebrates. The overlap of frequent fire spots with the richest regions of Zagros oak forests in terms of vertebrate diversity emphasizes the need for strategic forest risk reduction planning, especially in these priority zones.

Enigmatic vertebrate swimming trace fossils from the Wapiti Formation, Alberta, Canada, and their implications for paleoenvironmental reconstruction

Vertebrate ichnofossil research, when compared with invertebrate ichnological analyses, tends to put greater emphasis on identifying the trace makers rather than associating the trace fossils with animal behaviors to interpret local paleoenvironmental conditions. In 2011, several sandstone blocks bearing unusual deformational structures were collected from the Wapiti Formation exposed near Red Willow Falls, Alberta, Canada. The sandstone blocks were analyzed for their sedimentary characteristics and the morphology of the enigmatic structures in order to identify their origins and their significance for paleoenvironmental interpretation. The deformational features are interpreted to be the swim tracks of small vertebrates attempting to escape a flash flood event. The sedimentology of the sandstone suggests a rapid increase in water flow in an overbank setting, and the orientation of the traces indicate that the trace maker was initially moving against, and then moving perpendicular to, the current. This is likely the first case of a trace fossil exhibiting clear association between vertebrates and flash flood events. As such, these traces provide an excellent example of the utility of vertebrate trace fossils for assessing organism response to changing environmental conditions, thus facilitating interpretation of local geological events.

HIGH FREQUENCY OF BATS IN THE DIET OF THE BARN OWL TYTO ALBA IN A LOWLAND DRY FOREST IN TOLIMA, COLOMBIA

The cosmopolitan distribution of the Barn Owl Tyto alba and its relatively well documented diet through the study of pellets have provided evidence of the broad diversity of prey it relies on. In most studies, both in tropical and temperate regions, rodents are its main prey, with other small mammals and vertebrates, and insects complementing its diet. Although bats have been recorded in the diet of T. alba at many localities, they seldom represent an important part of its diet. This study describes the diet of a small colony of T. alba from a mosaic of agricultural fields, pastures for cattle ranching and tropical dry forests in Tolima, central Colombia. Overall, we collected 516 pellets between 2016 and 2017 in a non-systematic manner, and recovered 335 skulls of rodents (67%), bats (29%), birds (3%) and shrews (1%), as well as invertebrate prey. The large proportion of bats in the diet of T. alba in this study is only matched by a few other studies, and further documents the wide range of prey included in the diet of this nocturnal raptor. This is one of the first studies on the diet of T. alba in tropical dry forests in Colombia and complements the existing data on its diet in the American tropics and worldwide.

First Broad-Range Serological Survey of Crimean-Congo Hemorrhagic Fever among Hungarian Livestock.

(1) Background: Crimean-Congo hemorrhagic fever (CCHF) is an emerging tick-borne disease endemic in Africa, Asia, the Middle East, and the Balkan and Mediterranean regions of Europe. Although no human CCHF cases have been reported, based on vector presence, serological evidence among small vertebrates, and the general human population, Hungary lies within high evidence consensus for potential CCHF introduction and future human infection. Thus, the aim of our pilot serosurvey was to assess CCHF seropositivity among cattle and sheep as indicator animals for virus circulation in the country. (2) Methods: In total, 1905 serum samples taken from free-range cattle and sheep in 2017 were tested for the presence of anti-CCHF virus IgG antibodies using commercial ELISA and commercial and in-house immunofluorescent assays. (3) Results: We found a total of eleven reactive samples (0.58%) from five administrative districts of Hungary comprising 8 cattle and 3 sheep. The most affected regions were the south-central and northwestern parts of the country. (4) Conclusions: Based on these results, more extended surveillance is advised, especially in the affected areas, and there should be greater awareness among clinicians and other high-risk populations of the emerging threat of CCHF in Hungary and Central Europe.

Wombat burrows are hotspots for small vertebrates in a landscape subject to gigafire.

Ecosystem engineers modify their environment and influence the availability of resources for other organisms. Burrowing species, a subset of allogenic engineers, are gaining recognition as ecological facilitators. Burrows created by these species provide habitat for a diverse array of other organisms. Following disturbances, burrows could also serve as ecological refuges, thereby enhancing ecological resistance to disturbance events. We explored the ecological role of Common Wombat (Vombatus ursinus) burrows using camera traps in forests of southeastern Australia. We compared animal activity at paired sites with and without burrows, from the same fire severity class and habitat. We examined how animal activity at Common Wombat burrows was affected by the 2019-20 Black Summer bushfires in Australia. We predicted that burrows would serve as hotspots for animal activity and as refuges in burned areas. The activity of several species including Bush Rat (Rattus fuscipes), Agile Antechinus (Antechinus agilis), Lace Monitor (Varanus varius), Painted Button-quail (Turnix varius), and Grey Shrike-thrush (Colluricincla harmonica) increased at sites where Common Wombat burrows were present, while other species avoided burrows. Species that were more active at burrows tended to be smaller mammal and bird species that are vulnerable to predation, whereas species that avoided burrows tended to be larger mammals that might compete with Common Wombat for resources. Species composition differed between sites with and without burrows, and burrow sites had higher native mammal species richness. The association of several species with burrows persisted or strengthened in areas that burned during the 2019-20 Black Summer bushfires, suggesting that Common Wombat burrows may act as ecological refuges for animals following severe wildfire. Our findings have relevance for understanding how animals survive, persist, and recover following extreme wildfire events.

Planning for the future: Grasslands, herbivores, and nature‐based solutions

Abstract Global interest and investment in nature‐based solutions (NbS) are rapidly increasing because of the potential of this approach to concurrently counter biodiversity loss, provide cost‐effective measures for climate change adaptations, and maintain natural processes that underpin human health and wellbeing. Recognition is growing that grasslands in many regions will protect carbon stores more effectively than forests in the warmer, drier, more fire‐prone conditions of the future while also serving as hotspots for biodiversity. Yet grasslands have received less attention for their NbS potential. Despite the wide‐ranging goals of this approach, many investments in nature‐based solutions also have focused narrowly on using plants to meet climate pledges, often without considering plant interactions with herbivores and the abiotic environment that jointly control ecosystem functioning and underpin the success of nature‐based solutions. Here, we review the roles that large and small vertebrate and invertebrate herbivores play in the ability of the world's grasslands to provide nature‐based solutions, with a focus on wild herbivore impacts on biodiversity and carbon storage. Synthesis. Planning for nature‐based solutions with a holistic, ecologically informed view that includes the role of herbivores and their interaction with plants and the environment will allow NbS investments to more likely achieve successful, sustainable outcomes.

Carrion type and extent of breeding success together influence subsequent carrion choice by adult burying beetles

Abstract Insects can adjust their behaviour in relation to experience in a wide range of contexts including foraging, mate selection and choice of oviposition site. Here, we investigate whether burying beetles Nicrophorus vespilloides modulate their choice of carrion type in relation to the outcome of their past breeding experience. Burying beetles require the carcass of a small vertebrate, such as a mouse or a chick, to reproduce. Beetle parents convert this carrion into an edible nursery for their larvae, whom they typically care for throughout larval development. We tested whether a beetle's past breeding experience influenced its subsequent choice of carrion when presented simultaneously with either a dead mouse or a dead chick. We found that beetles that bred on a mouse and produced many larvae in their first breeding attempt subsequently favoured mice over chicks for their second breeding attempt. Beetles that had produced fewer larvae on a dead mouse switched to favouring dead chicks in their second breeding attempt. Those that had bred on a dead chick chose carrion at random subsequently, regardless of their previous breeding success. Our general conclusion is that burying beetles can integrate different sources of information about their past breeding experience with current cues when selecting carrion for reproduction.

A new species of the genus Paradistomum (Platyhelminthes: Trematoda) from Iwasaki's snail-eating snake Pareas iwasakii, with a note on morphological variations of Paradistomum megareceptaculum (Tamura, 1941).

Dicrocoeliid trematodes were detected from Iwasaki's snail-eating snake Pareas iwasakii in Iriomote Island, Okinawa Prefecture, Japan, and described as a new species Paradistomum dextra n. sp. in the present study. This new species can be distinguished from the type series of the other members of the genus based on size of eggs and morphological characteristics of body, oral and ventral suckers, and reproductive organs. However, the new species was hard to distinguish from Paradistomum megareceptaculum infecting snakes in Japan, including Iriomote Island where is the type locality of the new species, because it is closely similar to some part of the broad range of morphological variations in P. megareceptaculum. On the other hand, a partial sequence of 28S ribosomal DNA clearly distinguished these two species. Moreover, the new species' host snake Pareas iwasakii is reported to exclusively feed on land snails while host snakes of P. megareceptaculum feed on small vertebrates, indicating that the new species is also ecologically different from P. megareceptaculum. We also redescribed P. megareceptaculum based on adults sampled in this study and past studies to record the morphological variations of this species.

New occurrences of mammals from McKay Reservoir (Hemphillian, Oregon)

AbstractEncompassing global cooling, the spread of grasslands, and biogeographic interchanges, the Hemphillian North American Land Mammal Age is an important interval for understanding the factors driving ecological and evolutionary change through time. McKay Reservoir near Pendleton, Oregon is a natural laboratory for analyses of these factors. It is remarkable for its small vertebrate fauna including rodents, bats, turtles, and lagomorphs, but also for its larger mammal fossils like camelids, rhinocerotids, canids, and felids. Despite the importance of the site, few revisions to its faunal list have been published since its original description. We expand on this description by identifying taxa not previously known from McKay Reservoir based on specimens collected during fieldwork and through reidentification of previously collected fossils. Newly identified taxa include the borophagine canid Borophagus secundus (Matthew and Cook, 1909), the camelids Megatylopus Matthew and Cook, 1909 and Pleiolama Webb and Meachen, 2004, a dromomerycid, and the equids Cormohipparion Skinner and MacFadden, 1977 and Pseudhipparion Ameghino, 1904. Specimens previously assigned to Neohipparion Gidley, 1903 and Hipparion de Christol, 1832 lack the features necessary to diagnose these genera, which are therefore removed from the site's faunal list. The presence of Borophagus secundus, Cormohipparion, and Pseudhipparion is especially important, because each occurrence represents a major geographic range extension. This refined understanding of the fauna lays the foundation for future studies of taphonomy, taxonomy, functional morphology, and paleoecology—potentially at the population, community, or ecosystem levels—at this paleobiologically significant Miocene locality.

Environmental colour pattern variation in Mediterranean Podarcis

BackgroundLizards of the genus Podarcis are widespread in the Mediterranean region, including islands and island archipelagos. These small-bodied lizards have a predominantly protective green-brown colouration. However, some populations display unusual patterns, in which the colouration is predominantly blue or uniformly black. This study explores the factors that influence this chromatic variation, whether environmental (climate and island conditions) or evolutionary (phylogenetic trait conservatism). The colouration of 1400 individuals (27 species) was analysed in the CIELAB colour space.ResultsPagel’s λ indicated that colouration is weakly conserved within phylogenetic lineages. Although the island surface plays a key role in the chromatic variability of these lacertids, geographic isolation and climate hold less influence. The colouration of some small island populations tends to be uniform and dark, possibly due to intense intraspecific competition and lower predatory pressure.ConclusionsThis study highlights the importance of island populations in understanding the processes that favour the emergence of extreme phenotypes in small ectothermic vertebrates.

Detection of extra‐pair maternity in a carrion beetle under natural conditions

Abstract The provision of care to offspring is a costly endeavour that can be hijacked by others—known as brood parasitism. Females can choose to lay eggs in the nest of a conspecific female, resulting in extra‐pair maternity (EPM). Burying beetles of the genus Nicrophorus use small vertebrate carcasses for reproduction and provide biparental care to their offspring. Carcasses are a rare resource; thus, competition often occurs among adults of the same sex. A previous study examined the outcomes of competition and subsequent parental care of Nicrophorus vespilloides using laboratory experiments. Larvae of losing females were sometimes cared for by the winning females, which is generally an evidence of EPM. However, the presence of EPM in burying beetles under natural conditions has not yet been well established. Here, we focused on the reproduction of N. quadripunctatus in the field. To examine the presence of EPM, maternity analysis was performed on 149 offspring and their caring females in 10 broods, using 8 microsatellite DNA loci. Seven larvae originating from three broods were unrelated to their caring females. There were one to three unrelated larvae in each of the three broods. Thus, EPM occurs in N. quadripunctatus reproduction under natural conditions.