Longitudinal study of parasite-induced mortality of a long-lived host: the importance of exposure to non-parasitic stressors.

Hosts face mortality from parasitic and environmental stressors, but interactions of parasitism with other stressors are not well understood, particularly for long-lived hosts. We monitored survival of flour beetles (Tribolium confusum) in a longitudinal design incorporating cestode (Hymenolepis diminuta) infection, starvation and exposure to the pesticide diatomaceous earth (DE). We found that cestode cysticercoids exhibit increasing morphological damage and decreasing ability to excyst over time, but were never eliminated from the host. In the presence of even mild environmental stressors, host lifespan was reduced sufficiently that extensive degradation of cysticercoids was never realized. Median host lifespan was 200 days in the absence of stressors, and 3-197 days with parasitism, starvation and/or DE. Early survival of parasitized hosts was higher relative to controls in the presence of intermediate concentrations of DE, but reduced under all other conditions tested. Parasitism increased host mortality in the presence of other stressors at times when parasitism alone did not cause mortality, consistent with an interpretation of synergy. Environmental stressors modified the parasite numbers needed to reveal intensity-dependent host mortality, but only rarely masked intensity dependence. The longitudinal approach produced observations that would have been overlooked or misinterpreted if survival had only been monitored at a single time point.

Response of Flour Beetles to Multiple Stressors of Parasitic (Hymenolepis diminuta), Environmental (Diatomaceous Earth), and Host (Reproduction) Origin.

Organisms face a multitude of potential stressors, and the way these stressors interact can provide insights into underlying biological processes. This study examined the flour beetle Tribolium confusum and its survival, net fecundity, and surface-seeking behavior in response to combinations of stressors from 3 categories. Infection by the cestode Hymenolepis diminuta provided a stress of parasitic origin. Exposure to diatomaceous earth (DE) provided a stress of environmental origin. Use of virgin and mated beetles evaluated reproduction as a stress of host origin. Single and multiple exposure of beetles to parasite eggs achieved a maximum mean abundance of 21 parasites/beetle and a maximum intensity of 90 parasites in an individual beetle. DE reduced initial parasite establishment, but did not directly affect survival of parasites after their establishment in the host. A rehydration technique was used to recover parasites from dead beetles, enabling this to be the first study to correlate H. diminuta intensity at time of death directly to mortality of T. confusum. A dichotomous intensity-mortality relationship was observed in 8% DE, whereby lightly infected (<20 parasites) hosts were killed by DE in an intensity-independent manner, but more heavily infected hosts were killed in an intensity-dependent manner. Host mating status did not affect host survival, but there were interactions among mating status, parasitism, and DE on net fecundity and surface-seeking behavior. However, these effects were minor compared to the host mortality that occurred when parasite abundance and DE concentration were both high. The aggregated distribution of T. confusum in beetles, the difficulty of achieving high mean abundances, and an apparent need for the stressors to have strong effects individually if they are to have enhanced effects when in combination, suggests that exposure to multiple stressors would seriously impact only a small proportion of the host population.

Sublethal exposure to diatomaceous earth increases net fecundity of flour beetles (Tribolium confusum) by inhibiting egg cannibalism.

Population regulation results from an interplay of numerous intrinsic and external factors, and for many insects cannibalism is such a factor. This study confirms a previously-reported observation that sublethal exposure to the fossilized remains of diatoms (i.e. diatomaceous earth) increases net fecundity (eggs produced minus eggs destroyed/day) of flour beetles, Tribolium confusum. The aim was to experimentally test two non-mutually-exclusive ecological mechanisms potentially responsible for the increased net fecundity: higher egg production and lower egg cannibalism. Adult T. confusum were maintained at low or high density in medium containing sublethal (0-4%) diatomaceous earth. Net fecundity increased up to 2.1× control values during diatomaceous earth exposure, and returned to control levels following removal from diatomaceous earth. Cannibalism assays on adults showed that diatomaceous earth reduced the number of eggs produced to 0.7× control values at low density and to 0.8× controls at high density, and also reduced egg cannibalism rates of adults to as little as 0.4× control values, but at high density only. Diatomaceous earth also reduced cannibalism by larvae on eggs to 0.3× control values. So, while the presence of diatomaceous earth reduced egg production, net fecundity increased as a result of strong suppression of the normal egg cannibalism by adults and larvae that occurs at high beetle density. Undisturbed cultures containing sublethal diatomaceous earth concentrations reached higher population densities than diatomaceous earth-free controls. Cohort studies on survival from egg to adult indicated that this population increase was due largely to decreased egg cannibalism by adult females. This is the first report of inhibition of egg cannibalism by diatomaceous earth on larval or adult insects. The ability of diatomaceous earth to alter cannibalism behavior without causing mortality makes it an ideal investigative tool for cannibalism studies.

Hymenolepis diminuta infections in tenebrionid beetles as a model system for ecological interactions between helminth parasites and terrestrial intermediate hosts: a review and meta-analysis.

The cestode Hymenolepis diminuta (Cyclophyllidea) uses a variety of insects as its intermediate host, where ingestion of eggs results in development in the hemocoel of a cysticercoid that is infective to a rat definitive host. Species in 2 genera, Tenebrio and Tribolium (Coleoptera: Tenebrionidae) have been used extensively as laboratory intermediate hosts. This review examines experimental studies on ecological aspects of the relationship between H. diminuta and tenebrionid beetles, including the acquisition and establishment of the parasite, host effects on the parasite, and parasite effects on the host. A meta-analysis of infection results from the literature revealed strong relationships across host species and strains between (1) prevalence and intensity of infection, (2) efficiency of cysticercoid production and exposure conditions, and (3) variance in abundance or intensity of infection relative to their respective means. The underlying mechanisms producing these patterns remain elusive. Comparative studies are infrequent, and the use of divergent methodologies hampers comparisons among studies. In spite of these problems, there is much to recommend this as a terrestrial host-parasite model system. It represents those relationships in which mostly minor, but occasionally major, responses to parasitic infection occur, and in which host genetics and environmental conditions can serve as modifying factors. Moreover, this is a tractable experimental system, and is backed by an extensive literature on host biology.

Sequential and concurrent exposure of flour beetles ( Tribolium confusum ) to tapeworms ( Hymenolepis diminuta ) and pesticide (diatomaceous earth).

The response of Tribolium confusum to sublethal levels of 2 environmental stressors was studied, i.e., parasitic infection represented by the cestode Hymenolepis diminuta , and a physical stressor represented by the natural pesticide diatomaceous earth (DE). These were applied sequentially (DE, then infection) to detect indirect or carryover effects of DE, and concurrently (DE applied immediately after exposure to parasites and DE presence maintained throughout the infection) to detect direct effects of DE. DE alone, but not parasitism alone, produced significant host mortality, and concurrent treatment with DE and parasitism did not increase mortality over DE alone. Parasite abundance was significantly higher following sequential, but not concurrent, DE exposure. Parasite abundance in mated hosts was significantly higher than in virgin hosts. Parasitic infection resulted in significantly fewer eggs retained in the oviduct of beetles, but there was no difference in the number of eggs that accumulated in the culture medium and no difference in the surface-seeking behavior of beetles. Mating status of beetles in all treatments, and DE exposure in concurrent treatments significantly increased their surface-seeking behavior. Concurrent exposure to DE also resulted in a 4- to 6-fold increase in host egg numbers that accumulated in the culture medium. Although DE exposure increased parasite numbers in the beetles, these 2 stressors otherwise appeared to act independently.

Hymenolepis diminuta (Cestoda) induces changes in expression of select genes of Tribolium confusum (Coleoptera).

The flour beetle Tribolium confusum is a common experimental intermediate host for the tapeworm Hymenolepis diminuta, but while many aspects of their host-parasite interactions have been determined to have genetic basis, the genes involved have not been identified. In this paper, we report on the expression of several predicted metabolic and defense-related genes using quantitative polymerase chain reaction 2 weeks after initial infection of the beetle. The expression of heat shock protein 68, a predicted sugar transporter, a pheromone binding protein, and endoglin were up-regulated in infected beetles. The expression of thaumatin-like protein and prophenoloxidase 2/3 was down-regulated in infected beetles, while the mRNA levels of Toll-like receptor 3, Toll-like receptor 4, and lysozyme 4 were not affected by infection with H. diminuta.

Effect of age of the intermediate host Tribolium confusum (Coleoptera) on infection by Hymenolepis diminuta (Cestoda).

A cross-sectional study of 27 cohorts of Tribolium confusum aged 2-78 wk was done to examine effects of host age on exposure to eggs of Hymenolepis diminuta under standardized conditions. Pre-exposure, fasting, and postexposure mortality were low, sex ratio was equal, and fecundity of hosts was high during the first 30 wk, followed by increasing mortality and male bias of the sex ratio, and declining fecundity, in older beetles. These changes in the host were not associated with pronounced changes in infection results. Prevalence of infection was higher in females than males, but was unaffected by age in both sexes. Intensity of infection was similar between sexes in beetles up to 30 wk old, and thereafter declined in females, but not in males. Age-related changes in hosts were gradual, but unexpected levels of short-term variation in infection results suggest that some undetermined proximate factors may override general host age effects on the infection process.

Shape variation of cysticercoids of Hymenolepis diminuta (Cyclophyllidea) from fed, partially fed, and fasted Tribolium confusum (Coleoptera).

Quantitative studies of a crowding effect on cysticercoids of Hymenolepis diminuta in the intermediate host are few and limited in scope. In this study, we developed a technique to rapidly collect morphological information on large numbers of parasites, and verified the utility of geometric models for simple and accurate estimation of cysticercoid size for quantitative studies. These models were tested using measurements from 4,899 H. diminuta obtained from 666 Tribolium confusum exposed 1-4 wk previously. Length, width, and depth of the body and cercomer (when present) can be used in conjunction with these models to provide the most accurate estimation of parasite size. However, parasite body length alone can be used, with adjustment for effects of host diet and infection intensity, to predict the remaining measurements in incomplete specimens. Parasites that developed in higher intensity infections, or in hosts with reduced food intake, were narrower and had a proportionately shorter cercomer. Host age, sex, and mating status, and parasite age also had statistically significant, but small-magnitude, effects on parasite shape.