Infected Beetles Research Articles

Two distinct host-parasite associations mediate seasonal ecosystem linkages.

Nematomorph parasites manipulate terrestrial arthropods to enter streams where the parasites reproduce. These manipulated arthropods become a substantial prey subsidy for stream salmonids, causing cross-ecosystem energy flow. Diverse nematomorph-arthropod associations underlie the energy flow, but it remains unknown whether they can mediate the magnitude and temporal attributes of the energy flow. Here, we investigated whether distinct phylogenetic groups of nematomorphs manipulate different arthropod hosts and mediate seasonal prey subsidy for stream salmonids. The results of our molecular-based diagnoses show that Gordionus and Gordius nematomorphs infected ground beetle and orthopteran hosts, respectively. The presumable ground beetle hosts subsidized salmonid individuals in spring, whereas the presumable orthopteran hosts did so in autumn. Maintaining the two distinct nematomorph-arthropod associations thus resulted in the parasite-mediated prey subsidy in both spring and autumn in the study streams. Manipulative parasites are common, and often associated with a range of host lineages, suggesting that similar effects of phylogenetic variation in host-parasite associations on energy flow might be widespread in nature.

The Eucalyptus snout beetle in Colombia: Selection and evaluation of entomopathogenic fungi as bioinsecticides against Gonipterus platensis

The Eucalyptus snout beetles, which belong to the Gonipterus scutellatus species complex, cause heavy damage to Eucalyptus spp. worldwide. The species was reported in Antioquia, Colombia for the first time in 2016, threatening more than 115,000 ha of planted Eucalyptus spp. Elsewhere, its damage has been controlled using Anaphes spp. as classical biological control, but Anaphes spp. are not available in Colombia, where there are no documented control methods for this species to date. We identified the Gonipterus species using molecular methods and isolated entomopathogenic fungi from naturally infected beetles collected from the current distribution area in Colombia. We characterised the fungal isolates, selecting the best to develop a biological control agent. We considered insecticidal activity, UV-B radiation tolerance, yield, germination, and enzyme activity of conidia in making our selection. We confirmed the presence of G. platensis in Colombia, with adults naturally infected by Beauveria spp. and Metarhizium spp. From the isolated native fungi, M. brunneum (CA-3), M. robertsii (RI-1), B. bassiana (CA-1) and (CA-2), B. pseudobassiana (SP-1) were identified. We found remarkable differences in response to different variables between the selected isolates and SP-1 and CA-3 were the most virulent. The SP-1 isolate stood out from the rest for being the most tolerant to UV-B radiation, alkaline media, and low temperatures, germinating faster and producing elevated levels of lipases, chitinases, and proteases. The identified promising native fungus isolate, naturally occurring on field individuals, could be used to develop a biopesticide for controlling G. platensis in Eucalyptus spp. plantations in Colombia.

Alkaloid content in microsporidia-infected Adalia bipunctata (Coleoptera: Coccinellidae) life stages, and pathogen spore load in adults after exposure to physical stress

The two-spotted lady beetle, Adalia bipunctata L., displays warning colouration that is reinforced by the production of adaline and adalinine. These alkaloids are thought to provide defense against predation throughout all life stages of A. bipunctata and may play a role in the insect immune system. Vairimorpha (Nosema) adaliae, a microsporidium described from A. bipunctata, has minimal effects on its host (delayed larval development) when reared under optimum conditions but stress factors are shown to affect the development of microsporidiosis. The objectives of this study were to determine the effects of V. adaliae on relative alkaloid content (adaline) during A. bipunctata development, and to evaluate the combined effects of physical stress and infection on adult beetles (relative alkaloid content and infection load). First-instar larvae were isolated from uninfected and V. adaliae-infected colonies. Eggs and first-instar larvae were immediately prepared for alkaloid analysis, whereas late-instar larvae, pupae and adults were systematically processed when each reached their designated developmental stage. Upon eclosion, a subsample of beetles was exposed to varying amounts of physical agitation: control (no shaking), alternate shaking (every other day), and daily shaking. Immediately following these stress trials, alkaloid samples were collected for analysis and spore loads were assessed. Overall, relative adaline proportions increased from egg to adult. Uninfected individuals had significantly higher relative proportions of adaline than did infected individuals during early development; however, adaline content was higher in infected A. bipunctata from the third-instar onwards, when compared to their uninfected counterparts. Following exposure to physical agitation on alternate days, uninfected adults had a significantly higher relative proportion of adaline than did infected adults. Interestingly, exposure to different levels of agitation had no significant effect on alkaloid production for either uninfected or infected beetles. Mean spore counts were significantly higher for adults that were exposed to daily shaking when compared to individuals from the control and alternate shaking groups. From a biological perspective, one would expect to observe differences in alkaloid production through coccinellid development, as each successive life stage faces different external pressures and risks. When infected with the microsporidium V. adaliae, however, adaline production was reduced during early development but increased significantly in late life stages.

Near‐optimal control of a stochastic pine wilt disease model with prevention strategies

Pine wood nematode disease has become nowadays a major threat to forest ecosystems and caused huge economic and environmental losses to many countries and regions. Considering the influence of environmental factors such as temperature, drought, and humidity, as well as possible prevention strategies, including trunk injection, cutting infected pines, and insecticide application, in this paper, we develop a stochastic pine wilt disease model with prevention strategies, aiming to obtain the conditions for its near‐optimal control. The near‐optimality problem is constructed by decreasing the infected pine forests and beetle population while minimizing the costs of prevention strategies. We first verify that, under certain convexity conditions, the near‐minimum condition of the Hamiltonian function is sufficient for the near‐optimal control. Then, by utilizing the spike variational technique, we derive the necessary conditions of the near‐optimal control. Finally, we present an example to illustrate the validity of the main theoretical results, which indicates that reasonable prevention strategies could effectively reduce pine wilt infection, and therefore, to some extent, could protect the pine forests from pine wilt disease and reduce economic losses caused by this disease.

Transmission dynamics and sensitivity analysis of pine wilt disease with asymptomatic carriers via fractal-fractional differential operator of Mittag-Leffler kernel

Pine wilt disease is caused by nematodes transmitted by pine sawyer beetles and is fatal for several pine species. The trees might be destroyed within a few months after being attacked, leads to major ecological and financial losses. In this article, we presented a model of pine wilt disease in the trees considering the interaction between nematodes, transmitting beetles with both asymptomatic and symptomatic pine trees. The disease dynamics is first displayed through a schematic diagram which is then transformed to non-linear coupled integer order ODEs through the law of mass action. The positivity, boundedness and equilibrium points has been analyzed and basic reproduction number is calculated through the next generation technique. Sensitivity analysis is also done for the most sensitive parameters which is also displayed through different figures and tables. The solution of the considered nonlinear fractal-fractal model has been obtained through numerical method via MATLAB software. Some numerical results have been obtained and it has been observed that the fractional model give us the more general results by considering different non-integer orders and the integer order results can be easily recovered. It is also noticed that reducing the interaction among the infected beetles and susceptible trees by killing the main source of the infection i.e., killing beetles can reduce the infection drastically, which are briefly discussed and conclusion has been drawn on the basis of the obtained results.

Description of Joyeuxiella pasqualei (Cestoda: Dipylidiidae) from an Italian domestic dog, with a call for further research on its first intermediate host.

Although Joyeuxiella pasqualei is frequently detected in cats from Mediterranean Europe, information on its biology is still scarce. This cestode is relatively less frequently reported in dogs, possibly because it is often misdiagnosed with the better-known Dipylidium caninum. The occurrence of J. pasqualei proglottids in a dog living in a closed environment triggered us to delve into the biology of this cestode by collecting biological samples from lizards and a road-killed cat. Two reptile species, Podarcis siculus (Lacertidae), and Tarentola mauritanica (Geckonidae) were also collected in the garden and its surroundings. In addition, experimental infections with eggs obtained from gravid proglottids were performed in laboratory mice, and Tenebrio molitor (Coleoptera: Tenebrionidae) beetles. Proglottids from the dog's feces and adult cestodes detected at necroscopy of a cat were morphologically identified as J. pasqualei. Two out of 13 T. mauritanica collected in the garden had natural infections of J. pasqualei cysts in the liver and attached to the intestine. All P. siculus lizards and experimentally infected rodents and beetles were negative. DNA sequences obtained from J. pasqualei showed the highest nucleotide similarities with Versteria sp., Echinococcus sp., Raillietina sonini, Taenia polyacantha and D. caninum. Data herein provided show the inability of rodents to become infected by direct ingestion of gravid proglottids, suggesting a need for an invertebrate first intermediate host in the life cycle. Thus, more research study is advocated to better understand the biology of J. pasqualei such as its first intermediate host and its mechanism of transmission in reptiles and rodents.

Examination of Pine Wilt Epidemic Model through Efficient Algorithm

Pine wilt is a dramatic disease that kills infected trees within a few weeks to a few months. The cause is the pathogen Pinewood Nematode. Most plant-parasitic nematodes are attached to plant roots, but pinewood nematodes are found in the tops of trees. Nematodes kill the tree by feeding the cells around the resin ducts. The modeling of a pine wilt disease is based on six compartments, including three for plants (susceptible trees, exposed trees, and infected trees) and the other for the beetles (susceptible beetles, exposed beetles, and infected beetles). The deterministic modeling, along with subpopulations, is based on Law of mass action. The stability of the model along with equilibria is studied rigorously. The authentication of analytical results is examined through well-known computer methods like Non-standard finite difference (NSFD) and the model's feasible properties (positivity, boundedness, and dynamical consistency). In the end, comparison analysis shows the effectiveness of the NSFD algorithm.

Epizootological Significance of Rice Weevil as a Mycobacterium Bovis Reservoir

This article presents materials on the role of the rice weevil (lat. Sitophilus oryzae) in the process of transmission of Mycobacterium of tuberculosis. The relevance of the study is conditioned by the rapid spread of tuberculosis around the world and the need to develop more advanced methods for diagnosing this disease, which, for its part, is impossible without expanding knowledge on all possible reservoirs of the causative agent of infection. Long-term research around the world has proven that insects are carriers of various microflora, including pathogenic ones. Therefore the purpose of study was to establish the epizootological role of the rice weevil as a reservoir for Mycobacterium bovis in the process of occurrence and spread of tuberculosis. The study determined the duration of transfer and isolation of Mycobacterium of tuberculosis in the external environment by infected beetles. The museum dissociative strain of Mycobacterium bovis (118 passage) was used for the study, the subject of the study was rice weevils. For the furtherance of this goal, the culture, microscopic, and statistical studies were conducted, and the viability of mycobacteria was determined after passages through the beetle body by evaluating colony-forming units. The epizootological role of the rice weevil in the development and spread of tuberculosis has been established. It is proved that the beetle can keep mycobacteria in its body for 50 days with a gradual decrease in their number and is then able to secrete the pathogen for another 30 days, contaminating environmental objects. When evaluating colony-forming units of mycobacteria in the dynamics of the experiment, a decrease in their viability after persistence through the body of beetles was revealed from 5.3*108 in 1 g of the original crop up to 1.4*108 in 1 g of crop that grew from beetle homogenisate for 30 days. These study results will allow developing and improving existing measures for the prevention of tuberculosis and prevent the introduction of this infection into safe territory

Transcriptome Analysis of the Japanese Pine Sawyer Beetle, Monochamus alternatus, Infected with the Entomopathogenic Fungus Metarhizium anisopliae JEF-197.

The Japanese pine sawyer (JPS) beetle, Monochamus alternatus Hope (Coleoptera: Cerambycidae), damages pine trees and transmits the pine wilt nematode, Bursaphelenchus xylophilus Nickle. Chemical agents have been used to control JPS beetle, but due to various issues, efforts are being made to replace these chemical agents with entomopathogenic fungi. We investigated the expression of immune-related genes in JPS beetle in response to infection with JEF-197, a Metarhizium anisopliae isolate, using RNA-seq. RNA samples were obtained from JEF-197, JPS adults treated with JEF-197, and non-treated JPS adults on the 8th day after fungal treatment, and RNA-seq was performed using Illumina sequencing. JPS beetle transcriptome was assembled de novo and differentially expressed gene (DEG) analysis was performed. There were 719 and 1953 up- and downregulated unigenes upon JEF-197 infection, respectively. Upregulated contigs included genes involved in RNA transport, ribosome biogenesis in eukaryotes, spliceosome-related genes, and genes involved in immune-related signaling pathways such as the Toll and Imd pathways. Forty-two fungal DEGs related to energy and protein metabolism were upregulated, and genes involved in the stress response were also upregulated in the infected JPS beetles. Together, our results indicate that infection of JPS beetles by JEF-197 induces the expression of immune-related genes.

The effect of synthetic female sex pheromone on the transmission of the fungus Metarhizium brunneum by male Agriotes obscurus click beetles

Autodissemination techniques can potentially be used to distribute insecticides, including microbial insecticides, to cryptic pests. This approach is reliant on the target insect either passing the pathogen passively to other insects or the pathogen cycling within the population after the initial host dies. Here we examine, in small scale experiments, whether male Agriotes obscurus click beetles passively transmit the spores of the fungus Metarhizium brunneum directly, or indirectly via the environment, and whether this is influenced by exposure to synthetic female pheromone. We found that the beetles did not avoid M. brunneum spores and that this behaviour was not affected by pheromone. Exposure to pheromone increased beetle movement and uptake of spores, but this did not result in an increase in infected beetles under our conditions. Beetles were able to transfer spores at high levels via environmental contamination. However, contamination of the environment declined rapidly after exposure to the spores. The results are discussed in the context of developing an autodissemination strategy for click beetles.

Probability of occurrence and phenology of pine wilt disease transmission by insect vectors in the Rocky Mountains

Abstract1. Pine wilt disease, caused by pinewood nematode (Bursaphelenchus xylophilus; abbreviated ‘PWN’), is a damaging and globally distributed insect‐vectored forest pathogen. Native forest tree mortality associated with PWN is newly reported from the Front Range of Colorado, but there is no regional information on PWN frequency or biology of local insect vectors, limiting management options.2. A sampling array was established to survey PWN in native pines (Pinus ponderosa) and longhorn beetles (Monochamus clamator and M. scutellatus) over 2 years and across natural and urban forest landscapes. We developed flight phenology models and evaluated effects of landscape factors on vector abundance and probability of infection.3. Flight phenology was similar for vectors; Monochamus flight initiated in mid‐July and continued into October for both species. We report the first M. clamator–PWN association in the United States. PWN was distributed in the region at rates lower than reported from its putative native range: 3.6 and 4.2% of sampled pines and beetles, respectively, tested positive for PWN. Many host trees were outwardly asymptomatic; infection frequency in tree populations varied considerably and four epicentres of vector infectivity were identified.4. Epicentres varied in timing of anomalous infective vector frequency – some epicentres had high abundances of infected beetles early in the growing season whereas others had high abundances of infected beetles late in the growing season, though PWN‐positive beetles were captured at all sites. Monochamus populations were found primarily in natural forest stands but migrated to urban areas late in the growing season. The only landscape factor positively correlated with abundances of both Monochamus species was distance to previous wildfire.5. PWN epicentres in the southern Rocky Mountains exhibit specific temporal windows of vector activity that differ from proximal sites. Urban forests, where the disease was initially observed in the region, do not support vector populations. Our results suggest that natural forest landscapes in the region are important reservoirs of PWN, and vector populations are especially abundant near burned stands. Collectively, our findings are important for timing disease management activities appropriately and help to distinguish priority areas for mitigation efforts.

Transcription Profile and Genomic Variations of Oryctes Rhinoceros Nudivirus in Coconut Rhinoceros Beetles.

Oryctes rhinoceros nudivirus (OrNV) is a double-stranded DNA (dsDNA) virus which has been used as a biocontrol agent to suppress the coconut rhinoceros beetle (Oryctes rhinoceros) in Southeast Asia and the Pacific Islands. A new wave of O. rhinoceros incursions in Oceania is thought to be related to the presence of low-virulence isolates of OrNV or virus-tolerant haplotypes of beetles. In this study, chronically infected beetles were collected from Philippines, Fiji, Papua New Guinea (PNG), and the Solomon Islands (SI). RNA sequencing (RNA-seq) was performed to investigate the global viral gene expression profiles and for comparative genomic analysis of structural variations. Maximum likelihood phylogenic analysis indicated that OrNV strains from the SI and Philippines are closely related, while OrNV strains from PNG and Fiji formed a distinct adjacent clade. We detected several polymorphic sites with a frequency higher than 35% in 892 positions of the viral genome. Nonsynonymous mutations were detected in several hypothetical proteins and 15 nudivirus core genes, such as gp034, lef-8, lef-4, and vp91 We found limited evidence of variation in viral gene expression among geographic populations. Only a few genes, such as gp01, gp022, and gp107, were differentially expressed among different strains. Additionally, small RNA sequencing from the SI population suggested that OrNV is targeted by the host RNA interference (RNAi) response with abundant 21-nucleotide small RNAs. Some of these genomic changes are specific to the geographic population and could be related to particular phenotypic characteristics of the strain, such as viral pathogenicity or transmissibility, and this requires further investigation.IMPORTANCE Oryctes rhinoceros nudivirus has been an effective biocontrol agent against the coconut rhinoceros beetle in Southeast Asia and the Pacific Islands for decades. The recent outbreak of these beetles in many South Pacific islands has had a significant impact on livelihoods in the region. It has been suggested that the resurgence and spread of the pest are related to the presence of low-virulence isolates of OrNV or virus-tolerant haplotypes of beetles. We examined viral genomic and transcriptional variations in chronically infected beetles from different geographical populations. A high number of polymorphic sites among several geographical strains of OrNV were identified, but potentially only a few of these variations in the genome are involved in functional changes and can potentially alter the typical function. These findings provide valuable resources for future studies to improve our understanding of the OrNV genetic variations in different geographic regions and their potential link to virus pathogenicity.

Effects of food availability on microsporidiosis and alkaloid production in the two-spotted lady beetle, Adalia bipunctata L.

The effects of food availability and infection with the microsporidium Nosema adaliae on alkaloid production in the two-spotted lady beetle, Adalia bipunctata L., was examined. Alkaloid production (relative percent adaline in reflex-fluid) and pathogen load (spore counts) were quantified for both uninfected and N. adaliae-infected A. bipunctata. Alkaloid content was significantly higher for beetles fed irregularly than for those fed daily. For beetles infected with N. adaliae, spore counts were significantly higher for those fed irregularly compared to those fed daily. These results suggest that adaline content in reflex-fluid is influenced by infection and that irregular food supply increases pathogen load.

Parasites modulate the gut-microbiome in insects: A proof-of-concept study.

Host-parasite interactions may be modulated by host- or parasite-associated microbes, but the role of these are often overlooked. Particularly for parasites with intestinal stages (either larval or adult), the host gut microbiome may play a key role for parasite establishment; moreover, the microbiome may change in response to invading parasites. Hypothesis testing at the organismal level may be hampered, particularly in mammalian definitive hosts, by ethical, logistical, and economical restrictions. Thus, invertebrates naturally serving as intermediate hosts to parasites with complex life cycles may inform the development of mammalian models as an early-stage host-parasite model. In addition, several important pathogens are vectored by insects, and insect gut microbiome-pathogen interactions may provide essential base-line knowledge, which may be used to control vectorborne pathogens. Here, we used the grain beetle, Tenebrio molitor, a host of the tapeworm Hymenolepis diminuta, to explore interactions between infection status and resident gut microbiota at two pre-determined time points (day two and seven) post infection. Using 16S/18S microbial profiling, we measured key parameters of the composition, relative abundance, and diversity of the host gut bacteriome and mycobiome. In addition, we quantified the systemic beetle immune response to infection by Phenoloxidase activity and hemocyte abundance. We found significant changes in the gut bacteriome and mycobiome in relation to infection status and beetle age. Thus, the relative abundance of Proteobacteria was significantly higher in the gut of infected beetles and driven mostly by an increased abundance of Acinetobacter. In addition, the mycobiome was less abundant in infected beetles but maintained higher Shannon diversity in infected compared with non-infected beetles. Beetles treated with a broad-spectrum antibiotic (Tetracycline) exhibited significantly reduced parasite establishment compared with the untreated control group, indicating that the host microbiome may greatly influence hatching of eggs and subsequent establishment of H. diminuta larvae. Our results suggest that experimental work using invertebrates may provide a platform for explorative studies of host-parasite-microbe interactions and their underlying mechanisms.

Zombie-Ant Fungi Emerged from Non-manipulating, Beetle-Infecting Ancestors.

The manipulation of animal behavior by parasitic organisms is one of the most complex adaptations to have arisen via natural selection. Among the most impressive examples of behavioral manipulation are the zombie-ant fungi [1]. In this association, ants are controlled to leave the colony and perform a stereotyped death grip behavior, where they bite onto vegetation over foraging trails, before being killed for the post mortem fungal growth. Manipulation functions to provide a platform outside the nest, from which fungal parasites actively shoot out spores, targeting foraging ants because within colony transmission is prevented by strong social immunity exhibited by social insect societies [2, 3]. It is not clear how such complex examples of host manipulation arose. To address this, we performed a broad-scale phylogenetic reconstruction of the order Hypocreales, to which the zombie-ant fungi, Ophiocordyceps, belong. In order to understand the patterns of host association and host switching along the evolution of Ophiocordyceps, we performed ancestral character state reconstruction analysis. We found that zombie-ant fungi likely arose from an ancestor that infected beetle larvae residing in soil or decaying wood, similar to extant beetle-infecting Ophiocordyceps species. Surprisingly, the jump led to an extensive species radiation observed after the development of behavioral manipulation. We suggest that the jump from solitary beetle larva to ants within a colony exposed the fungus to the robust social immunity of ant societies.

The potential of Beauveria bassiana inoculum formulated into a polymeric matrix for a microbial control of spruce bark beetle

ABSTRACTTwo local strains of Beauveria bassiana originally isolated from naturally infected spruce bark beetles in Slovakia were tested for their virulence to Ips typographus (IT) and for their compatibility with a polymeric matrix composed of low-molecular polyethylene. Conidia could be homogenously immobilized in the low-molecular polyethylene matrix with no adverse effect on their viability and infectivity. At constant temperature (25°C), viability of immobilized conidial decreased only by 1–2% after 7 or 14 days when compared with non-formulated conidia. In field conditions, viability of conidia formulated in the matrix was even significantly higher than non-formulated conidia 35 days after their application in traps. Conidia incorporated into the polymeric matrix were infective to IT adults in laboratory bioassays. Mean values of LC50 for native conidia (0.72–2.05 × 106 conidia ml−1) and conidia immobilized in the polymeric matrix (0.64–1.03 × 105 conidia mm−2) demonstrated high virulence. The efficacy of the local strains was significantly higher than that of B. bassiana strains from mycoinsecticides (Boverol®, Botanigard® ES and Naturalis-L®). Results showed potential of this polymeric material for its use in microbial control of IT when mixed with conidia of B. bassiana.

Spiroplasma infection in Harmonia axyridis - Diversity and multiple infection.

The heritable endosymbiotic bacterium Spiroplasma is found in the harlequin ladybird Harmonia axyridis. The proportion of beetles infected with Spiroplasma in different native H. axyridis populations varies from 2% to 49%. We investigated the polymorphism of Spiroplasma strains in samples from individual beetles from Kyoto, Vladivostok, Troitsa Bay, Novosibirsk, and Gorno-Altaisk. To identify Spiroplasma strains, we analyzed nucleotide polymorphisms of the 16S rRNA gene and the ribosomal internal transcribed spacer (ITS1). The majority of infected beetles were infected with two or more Spiroplasma strains. We measured Spiroplasma density in beetles with different infection status using quantitative PCR. The abundance of Spiroplasma in samples with a single infection is an order of magnitude lower than in samples with multiple infections. Density dependent biological effects of Spiroplasma are discussed.

Experimental porcine cysticercosis using infected beetles with Taenia solium eggs

Beetles are intermediate hosts for human and animal parasites, and several beetle species have been shown to carry Taenia eggs. An experimental porcine cysticercosis infection model was developed using beetles (Ammophorus rubripes) infected with Taenia solium eggs and then using these beetles for oral pig challenge. A total of 18 three months-old Landrace pigs were divided in four groups. Pigs from groups 1, 2, and 3 (n = 6 pigs per group) were challenged with one, three, and six beetles infected with T. solium eggs, containing approximately 52, 156 or 312 eggs respectively. Pigs were necropsied 12 weeks after infection to assess the presence of T. solium metacestode. Porcine cysticercosis by T. solium was produced in 17 out of 18 pigs (94.4%) challenged with infected beetles, all infected pigs had viable cysts. Only one pig from group 1 was negative to the presence of cysts. The median number of metacestodes per pig in groups 1, 2, and 3 were 2 (range 0–71), 26 (range 5–33) and 40 cysts (range 4–111), respectively. Experimental porcine cysticercosis infection is consistently obtained using beetles as mechanical vectors for T. solium eggs.

The influence of microsporidian pathogens from commercially available lady beetles on larval development of the green lacewing, Chrysoperla carnea, in the absence of infection.

In North America, more than 70 species of natural enemies are available for pest control, including the aphid predators, Adalia bipunctata L. (two-spotted lady beetle) and Hippodamia convergens Guérin-Méneville (convergent lady beetle), and the generalist predator Chrysoperla carnea Stephens (green lacewing). The two lady beetle species are known to host microsporidian pathogens: Nosema adaliae was originally described from Adalia bipunctata and Tubulinosema hippodamiae from H. convergens. Microsporidia are spore-forming pathogens that typically produce chronic, debilitating disease. Because the spores of both pathogens are transovarially transmitted through beetle eggs, the predation behavior of lacewing larvae provides an opportunity for the transmission of these pathogens when infected lady beetles and lacewings share the same local environment. In this study, uninfected and microsporidia-infected eggs from A. bipunctata and H. convergens were offered to C. carnea larvae. The development of larvae that consumed N. adaliae-infected eggs was not affected, but larval development was prolonged by almost 3 days for those that consumed two or more T. hippodamiae-infected eggs. Prolonged larval development is considered to be costly because larvae remain vulnerable to cannibalization by sibling larvae or other predators. Longevity did not differ significantly between sexes of C. carnea, and the sex ratio of newly eclosed adults did not differ from the previously reported sex ratio of 1♂: 1♀. Upon examination by light microscopy at the end of the trial, two C. carnea larvae were infected with N. adaliae and none were infected with T. hippodamiae, suggesting that T. hippodamiae influenced lacewing larval development without establishing an infection.

Comparative Proteomic Analysis of Hymenolepis diminuta Cysticercoid and Adult Stages.

Cestodiases are common parasitic diseases of animals and humans. As cestodes have complex lifecycles, hexacanth larvae, metacestodes (including cysticercoids), and adults produce proteins allowing them to establish invasion and to survive in the hostile environment of the host. Hymenolepis diminuta is the most commonly used model cestode in experimental parasitology. The aims of the present study were to perform a comparative proteomic analysis of two consecutive developmental stages of H. diminuta (cysticercoid and adult) and to distinguish proteins which might be characteristic for each of the stages from those shared by both stages. Somatic proteins of H. diminuta were isolated from 6-week-old cysticercoids and adult tapeworms. Cysticercoids were obtained from experimentally infected beetles, Tenebrio molitor, whereas adult worms were collected from experimentally infected rats. Proteins were separated by GeLC-MS/MS (one dimensional gel electrophoresis coupled with liquid chromatography and tandem mass spectrometry). Additionally protein samples were digested in-liquid and identified by LC-MS/MS. The identified proteins were classified according to molecular function, cellular components and biological processes. Our study showed a number of differences and similarities in the protein profiles of cysticercoids and adults; 233 cysticercoid and 182 adult proteins were identified. From these proteins, 131 were present only in the cysticercoid and 80 only in the adult stage samples. Both developmental stages shared 102 proteins; among which six represented immunomodulators and one is a potential drug target. In-liquid digestion and LC-MS/MS complemented and confirmed some of the GeLC-MS/MS identifications. Possible roles and functions of proteins identified with both proteomic approaches are discussed.