Horned Passalus Beetle Research Articles

Sex-specific effects of a parasite on stress-induced freezing behavior in a natural beetle-nematode system.

Some animals react to predation threats or other stressors by adopting a freezing posture in an attempt to avoid detection, and the duration of this behavior usually corresponds with individual personality, such that timid individuals freeze longer. Despite decades of research on this or related behaviors (thanatosis), never has the impact of parasitism been considered. Parasites could prolong the duration, if hosts are less motivated to move (i.e. lethargic), or they could reduce it, if hosts are motivated to forage more to compensate for energy drain. We examined this behavior within a natural beetle-nematode system, where hosts (horned passalus beetles, Odontotaenius disjunctus) are parasitized by a nematode, Chondronema passali. We exposed beetles (n = 238) to four stressors in our lab, including noise, vibration, light and inversion, and recorded how long they adopt a frozen stance. Afterward, we determined nematode burdens, which can range from dozens to hundreds of worms. Beetles tended to freeze for 20 seconds on average, with some variation between stressors. We detected no effect of beetle mass on the duration of freezing, and this behavior did not differ in beetles collected during the breeding or non-breeding season. There was a surprising sex-based difference in the impact of nematodes; unparasitized females remained frozen twice as long as unparasitized males, but for beetles with heavy nematode burdens, the opposite was true. From this we infer that heavily parasitized females are more bold, while males with heavy burdens would be more timid. The explanation for this finding remains elusive, though we can rule out many possibilities based on prior work on this host-parasite system.

Walking while Parasitized: Effects of a Naturally-Occurring Nematode on Locomotor Activity of Horned Passalus Beetles

Internal parasites typically are associated with a range of negative effects on their hosts, including reduced energy, which can manifest in behavioral alterations. With this in mind, we examined effects of a naturally-occurring nematode parasite, Chondronema passali, on locomotor activity level in horned passalus beetles, Odontotaenius disjunctus from Georgia, USA. This parasite is not well-studied but can number in the thousands in severely parasitized hosts. Prior study in our lab revealed that parasitized beetles actually consume more wood than unparasitized ones do, leading us to ask here, if parasitized beetles are also more physically active. Beetles were collected from nearby forests and housed individually in our lab. We created a simple tabletop arena to observe beetle locomotor activity, which was gridded and included small stones and paper objects. We allowed individual beetles to traverse the arena for 5 min and recorded the number of grid squares crossed. Then, beetles were dissected to determine parasite presence and level of infection (on a categorical scale). A total of 140 beetles were examined across three collections. Statistical analyses of locomotor activity revealed parasite severity predicted locomotor activity, but paradoxically, lightly-infected beetles were twice as active as those without this nematode. Activity diminished with increasing worm burdens thereafter, but even the group with the most severe burdens did not move less than those with no worms. From these results we conclude that this parasite does not result in overall reduction in activity, but rather it appears to come with heightened locomotion. Alternatively, this result could stem from the fact that more active beetles are simply more likely to contract the parasite.

Phylogeography and demographic expansion in the widely distributed horned passalus beetle, Odontotaenius disjunctus (coleoptera: Passalidae)

Dynamic climatic oscillations during the Pleistocene had profound effects on the distribution of species across North America. Although the role of historical climate change on speciation remains controversial, the impact on genetic variation within species has been well documented. Analyses of mtDNA sequences from the cytochrome oxidase I gene (911 bp) for 115 individuals of Odontotaenius disjunctus was combined with ecological niche modelling (ENM) to infer the demographic and population differentiation scenarios under present and past conditions. We inferred three lineages that diverged during the Pleistocene and replace each other geographically across the eastern United States. One of these lineages traverses previously identified genetic barriers for terrestrial animals including the Mississippi and Apalachicola Rivers and the Appalachian Mountains. We observed overlapping ranges between two haplotype groups as well as a region of secondary contact associated with ecological transition zone in northern Florida. The two continental lineages depict a genetic signature of a recent population increase associated with expanding niche envelope, whereas the clade restricted to peninsular Florida shows stable populations in a shrinking niche envelope. Given the lack of ecological separation, overlapping distribution of haplogroups and the presence of secondary contact zones, the taxonomic status of these lineages must await robust testing using multilocus DNA data to assess species boundaries.

Healing while parasitized: impact of a naturally‐occurring nematode during energy‐intensive wound‐healing in a beetle

AbstractThe rigours of the daily lives of insects sometimes lead to minor injuries and wounds, which must be healed to avoid entry of pathogens and to resume normal function. Such healing requires energy, which must be diverted from other bodily reserves. What happens if energy reserves are already low, as would occur in individuals coping with internal parasites? This question is addressed in the presemt study, using horned passalus beetles (Odontotaenius disjunctus) and their naturally‐occurring nematode Chondronema passali. Oxygen consumption rates are tested at rest, as well as after an experimental wound is applied, to evaluate energy requirements of wound‐healing in parasitized and nonparasitized hosts. Furthermore, wound‐healing rates are visually tracked with a numerical scoring system to directly measure the cost of parasitism on healing. At rest, parasitized beetles show no elevation in respiration (oxygen consumption). After wounding, the oxygen consumption of parasitized beetles is 10% higher than that in nonparasitized beetles. Beetles with moderate‐ to heavy worm burdens have slower healing than those with few or no nematodes. These results show that this parasite carries little cost to the host during day‐to‐day activities, whereas, during times of immediate energy demand, there is a cost; hosts require more energy to repair wounds, and the wounds take longer to close. This conclusion leads to the question of whether this parasite is truly benign, and how many other apparently benign parasites, in insects or other animals, have similar ‘hidden’ effects.

Two Divergent Genetic Lineages within the Horned Passalus Beetle, Odontotaenius disjunctus (Coleoptera: Passalidae): An Emerging Model for Insect Behavior, Physiology, and Microbiome Research.

The horned passalus (Odontotaenius disjunctus) is one of the most extensively studied saproxylic beetles in the eastern United States. For several decades this species has been the subject of investigations into the behaviors associated with subsociality as well as physiological responses to stress, and, most recently, the composition of its gut microbiome has been closely examined. However, no published study to date has characterized this beetle’s broad-scale population genetic structure. Here, we conducted intensive geographic sampling throughout the southern Appalachian Mountains and surrounding areas and then assessed mitochondrial DNA (mtDNA) sequence variation among individuals. Unexpectedly, we discovered two divergent, yet broadly sympatric, mtDNA clades. Indeed, the magnitude of divergence between- vs. within-clades ranged from 5.9 to 7.5×, depending on the dataset under consideration, and members of the two lineages were often syntopic (i.e., found in the same rotting log). Given the potential implications for past and future studies on behavior, physiology, and the gut microbiome, we developed a simple cost-efficient molecular assay (i.e., polymerase chain reaction restriction fragment length polymorphism; PCR-RFLP) to rapidly determine mtDNA clade membership of O. disjunctus individuals. We suggest that the evolutionary processes that gave rise to the emergence and persistence of divergent sympatric lineages reported here warrant investigation, as this type of spatial-genetic pattern appears to be rare among southern Appalachian forest invertebrates.

Direct measurement of fight or flight behavior in a beetle reveals individual variation and the influence of parasitism.

How and to what degree an animal deals with potential threats is a fascinating topic that has been well-researched, particularly in insects, though usually not with the impact of parasites in mind. A growing body of work is showing how even benign parasites can affect, positively or negatively, their hosts’ physiological or behavioral reaction to threats. With this in mind we conducted an experiment using horned passalus beetles, Odontotaenius disjunctus that were naturally parasitized with a nematode Chondronema passali; we subjected beetles to simulated attacks (resembling rival fighting or predator attacks) and from videos of the encounters we quantified a suite of behaviors (antennae movement, aggressive posturing, threat displays, etc.), plus rates of alarm calls (stridulations) which all correspond to the “fight or flight” reaction. We obtained behavioral and parasite data from 140 beetles from two field collections, of which half had been housed in our lab for three weeks in conditions that would be stressful (little cover for burrowing). We observed a wide range of behaviors during the simulated attack procedure, from beetles offering little resistance to those which were extremely aggressive, though most beetles showed a moderate reaction. Alarm calling rates also varied, but surprisingly, these were not correlated with the magnitude of behavioral reactions. Also surprising was that stressful housing did not heighten the physical resistance during attacks, but did elevate alarm calling rate. Importantly, parasitized beetles had significantly reduced physical reactions to attack than those without nematodes (meaning their resistance to the attack was muted). The results concerning parasitism, coupled with prior work in our lab, indicate that the C. passali nematode depresses the hosts’ acute stress, or fight or flight, reaction (likely from its energetic cost), which may make hosts more susceptible to the very dangers that they are coping with during the stress events.

Delayed dispersal and prolonged brood care in a family-living beetle.

Delayed juvenile dispersal is an important prerequisite for the evolution of family-based social systems, such as cooperative breeding and eusociality. In general, young adults forego dispersal if there are substantial benefits to remaining in the natal nest and/or the likelihood of dispersing and breeding successfully is low. We investigate some general factors thought to drive delayed juvenile dispersal in the horned passalus beetle, a family-living beetle in which young adults remain with their families in their natal nest for several months before dispersing. Fine-scale population genetic structure indicated high gene flow between nest sites, suggesting that constraints on mobility are unlikely to explain philopatry. Young adults do not breed in their natal log and likely disperse before reaching breeding age, suggesting that they do not gain direct reproductive benefits from delayed dispersal. We also examined several ways in which parents might incentivize delayed dispersal by providing prolonged care to adult offspring. Although adult beetles inhibit fungal growth in the colony by manipulating both the nest site and deceased conspecifics, this is unlikely to be a major explanation for family living as both parents and adult offspring seem capable of controlling fungal growth. Adult offspring that stayed with their family groups also neither gained more mass nor experienced faster exoskeleton development than those experimentally removed from their families. The results of these experiments suggest that our current understanding of the factors underlying prolonged family living may be insufficient to explain delayed dispersal in at least some taxa, particularly insects.

The Heartrate Reaction to Acute Stress in Horned Passalus Beetles (Odontotaenius disjunctus) is Negatively Affected by a Naturally-Occurring Nematode Parasite.

There are many events in the lives of insects where rapid, effective stress reactions are needed, including fighting conspecifics to defend territories, evading predators, and responding to wounds. A key element of the stress reaction is elevation of heartrate (HR), for enhancing distribution of blood (hemolymph) to body compartments. We conducted two experiments designed to improve understanding of the insect stress reaction and how it is influenced by parasitism in a common beetle species (Odontotaenius disjunctus). By non-destructively observing heartbeat frequency before, during and after applying a stressor (physical restraint) for 10 min, we sought to determine: (1) the exact timing of the cardiac stress reaction; (2) the magnitude of heartrate elevation during stress; and (3) if the physiological response is affected by a naturally-occurring nematode parasite, Chondronema passali. Restraint caused a dramatic increase in heartrate, though not immediately; maximum HR was reached after approximately 8 min. Average heartrate went from 65.5 beats/min to a maximum of 81.5 (24.5% increase) in adults raised in the lab (n = 19). Using wild-caught adults (n = 77), average heartrates went from 54.9 beats/min to 74.2 (35.5% increase). When restraint was removed, HR declined after ~5 min, and reached baseline 50 min later. The nematode parasite did not affect baseline heartrates in either experiment, but in one, it retarded the heartrate elevation during stress, and in the other, it reduced the overall magnitude of the elevation. While we acknowledge that our results are based on comparisons of beetles with naturally-occurring parasite infections, these results indicate this parasite causes a modest reduction in host cardiac output during acute stress conditions.

Parasite Manipulation of Its Host's Physiological Reaction to Acute Stress: Experimental Results from a Natural Beetle-Nematode System.

All animals, whether vertebrate or invertebrate, must be capable of reacting to acute stressors, such as escaping from predators, and most do so with a suite of transient physiological changes that temporarily enhance survival. Some of these changes include mobilization of immune cells and increased cardiac output. A small but growing number of studies have begun to show that certain parasites appear capable of modifying such responses. We addressed this topic using a natural host and parasite system, that is, a nematode (Chondronema passali) that parasitizes horned passalus beetles, Odontotaenius disjunctus (family Passalidae), of the eastern United States. With a series of experiments, we sought to determine whether this parasite affects (1) the immune reaction to stress, (2) the output of stress-induced alarm calls, or (3) the increase in heart rate that occurs in response to acute stressors, with the stressors being mechanical or thermal. Results showed that hemocyte density increased after both stressors in nonparasitized beetles but did not increase in parasitized beetles. While mobilization of immune cells would enhance host immunity during stress, this would also be damaging to the nematode, so this scenario appears to benefit the parasite. We found no evidence that the nematode suppresses the overall reaction to stress (or prevents stress from occurring), since parasitized beetles did not differ from nonparasitized ones in alarm call rates or in heart beat frequency after exposure to mechanical stressors. Suppression of the host's normal immune reaction to stressful stimuli could translate to delayed or even reduced wound healing or pathogen resistance during these events. This project is a rare demonstration of parasite manipulation of host immune response to acute stress and should stimulate further investigations into the interactive nature of stress and parasites.

Observations ofSteinernemaNematode and Tachinid Fly Parasites in Horned Passalus Beetles,Odontotaenius disjunctus, from Georgia, U.S.A.

ABSTRACT: This report describes a series of observations made on 2 parasite species infecting a collection of horned passalus beetles (Odontotaenius disjunctus, n = 135), from Georgia, U.S.A., that were collected as larvae in the wild and were reared to metamorphosis in captivity. Tachinid fly maggots emerged from 15 beetle larvae (11%) and, when they completed their development into adults, we identified them as Zelia vertebrata, a known but little-studied parasitoid of O. disjunctus. We also observed infections by Steinernema nematodes (n = 11 cases, or 8%), which killed and consumed the carcasses and which produced thousands of infective juveniles in the rearing containers. Based on morphological measurements of infective juvenile and adult worms, these nematodes were consistent with Steinernema carpocapsae, members of which infest the surface of soils and which infect a wide range of insects. This report is the first to describe Steinernema infections in O. disjunctus, and our observations of Z. vert...

Effects of the Naturally Occurring Parasitic NematodeChondronema passaliLeidy on Lifting Strength and Captivity-Related Body Mass Patterns in the Horned Passalus Beetle,Odontotaenius disjunctus(Illiger) (Coleoptera: Passalidae)

Abstract Parasites are ubiquitous among insects and well-studied, but knowledge of their full range of host effects is not complete, especially concerning their impact on physical performance of hosts. The horned passalus beetle, Odontotaenius disjunctus (Illiger), in the USA is frequently parasitized by Chondronema passali Leidy, a nematode that can number in the thousands within the hemocoel. We performed an experiment that examined the effects of this parasite on a key performance metric, lifting strength, which should be important during fighting. We assessed beetle lifting strength using a vertically-mounted force gauge, as well as their body mass. At the end of the experiment, we dissected the beetles to determine parasite status and gender. The average beetle (1–2 g each) lifted between 300 and 400 g during the trials. Though the final sample size of non-parasitized beetles was small (n = 8), our results showed parasitized beetles tended to lift 17% less weight than non-parasitized beetles, and thi...

Fighting while parasitized: can nematode infections affect the outcome of staged combat in beetles?

The effects of non-lethal parasites may be felt most strongly when hosts engage in intense, energy-demanding behaviors. One such behavior is fighting with conspecifics, which is common among territorial animals, including many beetle species. We examined the effects of parasites on the fighting ability of a saproxylic beetle, the horned passalus (Odontotaenius disjunctus, Family: Passalidae), which is host to a non-lethal nematode, Chondronema passali. We pitted pairs of randomly-chosen (but equally-weighted) beetles against each other in a small arena and determined the winner and aggression level of fights. Then we examined beetles for the presence, and severity of nematode infections. There was a non-significant tendency (p = 0.065) for the frequency of wins, losses and draws to differ between beetles with and without C. passali; non-parasitized individuals (n = 104) won 47% of their fights while those with the parasite (n = 88) won 34%, a 13% difference in wins. The number of nematodes in a beetle affected the outcome of fights between infected and uninfected individuals in an unexpected fashion: fighting ability was lowest in beetles with the lowest (p = 0.033), not highest (p = 0.266), nematode burdens. Within-fight aggression was highest when both beetles were uninfected and lowest when both were infected (p = 0.034). Collectively, these results suggest the nematode parasite, C. passali, is associated with a modest reduction in fighting ability in horned passalus beetles, consistent with the idea that parasitized beetles have lower energy available for fighting. This study adds to a small but growing body of evidence showing how parasites negatively influence fighting behavior in animals.

Lifting Capacity of Horned Passalus Beetles During Passive and Stressed States

Many beetle species engage in territorial behaviors or male-male contests involving lifting or flipping their opponents, although this type of strength has never been empirically quantified. This study examined the lifting capacity of a medium-sized (1–2 g) saprolytic beetle native to the United States (horned passalus beetle, Odontotaenius disjunctus), and strength was measured during passive and stressed states. Twenty beetles were individually placed in a ‘push-up’ position with a force sensor on their backs and allowed to lift continuously for 2 min without manipulation, and then for another 2 min with a mild stress stimulus applied (tapping the elytra with a probe). The unmanipulated peak force readings during the first half were surprisingly high (up to 5 N, or 500 g), based on prior experiments examining the pulling strength of this species (indicating they can pull up to 100 g), but in nearly all beetles their peak lifting power in the stressed state increased by an average of 87 %. There was a positive relationship between strength measurements in both passive and stressed states. This appears to be the first empirical demonstration of the lifting capacity of a beetle, and these results also have considerable implications for the study of physical performance in beetles and other animals, especially in cases where maximum exertion data are of interest.

Measuring the Strength of the Horned Passalus Beetle,Odontotaenius disjunctus: Revisiting an Old Topic with Modern Technology

Over a century ago, a pioneering researcher cleverly devised a means to measure how much weight the horned passalus beetle, Odontotaenius disjunctus (Illiger) (Coleoptera: Passalidae), could pull using a series of springs, pulleys, and careful observation. The technology available in modern times now allows for more rigorous data collection on this topic, which could have a number of uses in scientific investigations. In this study, an apparatus was constructed using a dynamometer and a data logger in an effort to ascertain the pulling strength of this species. By allowing beetles to pull for 10 min, each beetle's mean and maximum pulling force (in Newtons) were obtained for analyses, and whether these measures are related was determined. Then, whether factors such as body length, thorax size, horn size, or gender affect either measure of strength was investigated. Basic body measurements, including horn size, of males versus females were compared. The measurements of 38 beetles (20 females, 18 males) showed there was no difference in overall body length between sexes, but females had greater girth (thorax width) than males, which could translate into larger muscle mass. A total of 21 beetles (10 females, 11 males) were tested for pulling strength. The grand mean pulling force was 0.14 N, and the grand mean maximum was 0.78 N. Despite the fact that beetles tended to pull at 20% of their maximum capacity most of the time, and that maximum force was over 5 times larger than the mean force, the 2 measures were highly correlated, suggesting they may be interchangeable for research purposes. Females had twice the pulling strength (both maximum and mean force) as males in this species overall, but when the larger thorax size of females was considered, the effect of gender was not significant. Beetle length was not a significant predictor of pulling force, but horn size was associated with maximum force. The best predictor of both measures of strength appeared to be thorax size. There are a multitude of interesting scientific questions that could be addressed using data on beetle pulling strength, and this project serves as a starting point for such work.

Dynamics of tracheal compression in the horned passalus beetle

Rhythmic patterns of compression and reinflation of the thin-walled hollow tubes of the insect tracheal system have been observed in a number of insects. These movements may be important for facilitating the transport and exchange of respiratory gases, but observing and characterizing the dynamics of internal physiological systems within live insects can be challenging due to their size and exoskeleton. Using synchrotron X-ray phase-contrast imaging, we observed dynamical behavior in the tracheal system of the beetle, Odontotaenius disjunctus. Similar to observations of tracheal compression in other insects, specific regions of tracheae in the thorax of O. disjunctus exhibit rhythmic collapse and reinflation. During tracheal compression, the opposing sides of a tracheal tube converge, causing the effective diameter of the tube to decrease. However, a unique characteristic of tracheal compression in this species is that certain tracheae collapse and reinflate with a wavelike motion. In the dorsal cephalic tracheae, compression begins anteriorly and continues until the tube is uniformly flattened; reinflation takes place in the reverse direction, starting with the posterior end of the tube and continuing until the tube is fully reinflated. We report the detailed kinematics of this pattern as well as additional observations that show tracheal compression coordinated with spiracle opening and closing. These findings suggest that tracheal compression may function to drive flow within the body, facilitating internal mixing of respiratory gases and ventilation of distal regions of the tracheal system.

The structure and musculature of the larval head and mouthparts of the horned Passalus beetle, Popilius disjunctus Illiger

The structure and musculature of the larval head and mouthparts of the horned Passalus beetle, Popilius disjunctus Illiger

The Stages of Development in the Embryology of the Horned Passalus Beetle, Popilius Disjunctus Illiger

The horned passalus beetle, Popilius disjunctus Illiger (the Passalus cornutus Fabricius of many authors), possesses many interesting characteristics which make this widespread species especially desirable for developmental studies. One of these advantageous characteristics is the large size of the eggs compared to those of other common insects. When laid, the egg measures 3.0 by 2.4 mm. and when ready to hatch, the egg has increased to 3.7 by 3.2 mm. This large size makes the eggs very easy to manipulate. A second advantageous feature is the fact that the incubation period of sixteen days is long enough to enable the investigator to collect many transition stages in the development of the egg and allows for more convenient and accurate studies than if the embryological course were exceedingly rapid. Lastly, this large beetle completes its whole life history from egg to imago in a single summer, and similarly, this favors developmental studies.