Jitka Vilímová

Host association drives genetic divergence in the bed bug, Cimex lectularius.

Genetic differentiation may exist among sympatric populations of a species due to long‐term associations with alternative hosts (i.e. host‐associated differentiation). While host‐associated differentiation has been documented in several phytophagus insects, there are far fewer cases known in animal parasites. The bed bug, Cimex lectularius, a wingless insect, represents a potential model organism for elucidating the processes involved in host‐associated differentiation in animal parasites with relatively limited mobility. In conjunction with the expansion of modern humans from Africa into Eurasia, it has been speculated that bed bugs extended their host range from bats to humans in their shared cave domiciles throughout Eurasia. C. lectularius that associate with humans have a cosmopolitan distribution, whereas those associated with bats occur across Europe, often in human‐built structures. We assessed genetic structure and gene flow within and among populations collected in association with each host using mtDNA, microsatellite loci and knock‐down resistance gene variants. Both nuclear and mitochondrial data support a lack of significant contemporary gene flow between host‐specific populations. Within locations human‐associated bed bug populations exhibit limited genetic diversity and elevated levels of inbreeding, likely due to human‐mediated movement, infrequent additional introduction events per infestation, and pest control. In contrast, populations within bat roosts exhibit higher genetic diversity and lower levels of relatedness, suggesting populations are stable with temporal fluctuations due to host dispersal and bug mortality. In concert with previously published evidence of morphological and behavioural differentiation, the genetic data presented here suggest C. lectularius is currently undergoing lineage divergence through host association.

The role of autophagy in the midgut epithelium of Eubranchipus grubii (Crustacea, Branchiopoda, Anostraca)

Eubranchipus grubii (Crustacea, Branchiopoda, Anostraca) is an omnivorous filter feeder whose life span lasts no more than 12 weeks. Adult males and females of E. grubii were used for ultrastructural studies of the midgut epithelium and an analysis of autophagy. The midgut epithelium is formed by columnar digestive cells and no regenerative cells were observed. A distinct regionalization in the distribution of organelles appears - basal, perinuclear and apical regions were distinguished. No differences in the ultrastructure of digestive cells were observed between males and females. Autophagic disintegration of organelles occurs throughout the midgut epithelium. Degenerated organelles accumulate in the neighborhood of Golgi complexes, and these complexes presumably take part in phagophore and autophagosome formation. In some cases, the phagophore also surrounds small autophagosomes, which had appeared earlier. Fusion of autophagosomes and lysosomes was not observed, but lysosomes are enclosed during autophagosome formation. Autophagosomes and autolysosomes are discharged into the midgut lumen due to apocrine secretion. Autophagy plays a role in cell survival by protecting the cell from cell death.

The ultrastructure of the midgut epithelium in millipedes (Myriapoda, Diplopoda).

The midgut epithelia of the millipedes Polyxenus lagurus, Archispirostreptus gigas and Julus scandinavius were analyzed under light and transmission electron microscopies. In order to detect the proliferation of regenerative cells, labeling with BrdU and antibodies against phosphohistone H3 were employed. A tube-shaped midgut of three millipedes examined spreads along the entire length of the middle region of the body. The epithelium is composed of digestive, secretory and regenerative cells. The digestive cells are responsible for the accumulation of metals and the reserve material as well as the synthesis of substances, which are then secreted into the midgut lumen. The secretions are of three types - merocrine, apocrine and microapocrine. The oval or pear-like shaped secretory cells do not come into contact with the midgut lumen and represent the closed type of secretory cells. They possess many electron-dense granules (J. scandinavius) or electron-dense granules and electron-lucent vesicles (A. gigas, P. lagurus), which are accompanied by cisterns of the rough endoplasmic reticulum. The regenerative cells are distributed individually among the basal regions of the digestive cells. The proliferation and differentiation of regenerative cells into the digestive cells occurred in J. scandinavius and A. gigas, while these processes were not observed in P. lagurus. As a result of the mitotic division of regenerative cells, one of the newly formed cells fulfills the role of a regenerative cell, while the second one differentiates into a digestive cell. We concluded that regenerative cells play the role of unipotent midgut stem cells.

Fine structure of the midgut epithelium in two Archaeognatha, Lepismachilis notata and Machilis hrabei (Insecta), in relation to its degeneration and regeneration

In two archaeognathans, Lepismachilis notata and Machilis hrabei, the midgut epithelium and processes of its regeneration and degeneration have been described at the ultrastructural level.In both analysed species, the midgut epithelium is composed of epithelial and regenerative cells (regenerative nests). The epithelial cells show distinct regionalization in organelles distribution with the basal, perinuclear, and apical regions being distinguished. Degeneration of epithelial cells proceeds in a necrotic way (continuous degeneration) during the entire life of adult specimens, but just before each moult degeneration intensifies. Apoptosis has been observed. Regenerative cells fulfil the role of midgut stem cells. Some of them proliferate, while the others differentiate into epithelial cells.We compared the organisation of the midgut epithelium of M. hrabei and L. notata with zygentoman species, which have just been described.

Does autophagy in the midgut epithelium of centipedes depend on the day/night cycle?

The midgut epithelium of two centipedes, Lithobius forficatus and Scolopendra cingulata, is composed of digestive, secretory and regenerative cells. In L. forficatus, the autophagy occurred only in the cytoplasm of the digestive cells as a sporadic process, while in S. cingulata, it occurred intensively in the digestive, secretory and regenerative cells of the midgut epithelium. In both of the species that were analyzed, this process proceeded in a continuous manner and did not depend on the day/night cycle. Ultrastructural analysis showed that the autophagosomes and autolysosomes were located mainly in the apical and perinuclear cytoplasm of the digestive cells in L. forficatus. However, in S. cingulata, the entire cytoplasm was filled with autophagosomes and autolysosomes. Initially the membranes of phagophores surround organelles during autophagosome formation. Autolysosomes result from the fusion of autophagosomes and lysosomes. Residual bodies which are the last stage of autophagy were released into the midgut lumen due to necrosis. Autophagy in the midgut epithelia that were analyzed was confirmed using acid phosphatase and mono-dansyl-cadaverine stainings.

Molecular evidence places the swallow bug genus Oeciacus Stål within the bat and bed bug genus Cimex Linnaeus (Heteroptera: Cimicidae)

The genera Cimex Linnaeus and Oeciacus Stål (Heteroptera: Cimicidae) are common haematophagous ectoparasites of bats or birds in the Holarctic region. Both their phylogenetic relationship and the systematics of the entire family previously were based on data from morphology and host relationships. Relationships among nine species of the genus Cimex and three species of the genus Oeciacus were analysed here using two mitochondrial and three nuclear genes. Cimex was shown to be paraphyletic with respect to Oeciacus. Oeciacus is thus proposed as a synonym of Cimex. The characteristic phenotype of Oeciacus results from the specific host association with different species of swallows (Hirundinidae). The morphological characters that have been used as diagnostic for the genera were shown to be valid and can be further used for determination at species level. The present analyses recovered the four traditional morphologically defined species groups of the genus Cimex. However, their relationships were poorly resolved – only the C. hemipterus group showed a well‐supported relationship to the C. pipistrelli group. The molecular differentiation within the Palaearctic C. pipistrelli and the Nearctic C. pilosellus species groups correlates with their karyotype differentiation. Furthermore, the analyses suggest poly‐ or paraphyly of the former genus Oeciacus. Either way this indicates there is a large amount of host‐associated phenotypic convergence in either bat‐ or bird‐associated groups of species. The probability of host choice and subsequent switch in Cimicidae are discussed and possible scenarios of the evolution of host association in species of Cimex are suggested.

Distribution and host relations of species of the genus Cimex on bats in Europe

Abstract. The species of the genus Cimex (Heteroptera: Cimicidae) are important ectoparasites of European bats. Unlike other ectoparasites, they are attached to the body of their host only when they need to feed, otherwise they stay in refugia in bat roosts. Consequently, they are often overlooked by bat specialists and in many countries they are either unknown or poorly characterized. This study reports results from thorough investigations of bat roosts of diverse bat species in a Northwest-Southeast transect across Europe: Czech and Slovak Republics, Hungary, Serbia and Bulgaria. The distribution of Cimex lectularius follows the synanthropic habitats of its principal hosts, Myotis myotis and M. emarginatus, both Mediterranean elements of the European fauna. The climate in natural roosts (i.e. caves) inhabited by these bats in southern areas appears to restrain the presence of cimicids. In central Europe, C. pipistrelli parasitizes, beside M. myotis, many crevice-dwelling bat species indigenous to the boreal zone. However, in southern Europe, it appears only in connection with Nyctalus noctula. C. lectularius was confirmed for five host bat species and newly recorded for Rhinolophus ferrumequinum, C. pipistrelli was confirmed for seven bat species and newly recorded for Myotis nattereri. The first record of C. emarginatus outside of its type locality and Myotis alcathoe as a new host are reported. The host preferences of the species of the genus Cimex are discussed.

Insect community on Jurinea cyanoides (Asteraceae), a plant species protected under NATURA 2000

Abstract The plant Jurinea cyanoides (L.) Reichenbach (Asteraceae), protected under the European Commission Directive on the conservation of natural habitats and of wild fauna and flora (92/43/EEC), is critically endangered in Central Europe. The centre of its continuous range of distribution is in Ukraine and in a part of European Russia. Natural isolated populations have refugial distribution in the Czech Republic and Germany. Interactions of insects from different feeding guilds (i.e., phytophages, pollinators, parasitoids, predators) with J. cyanoides were studied in central Ukraine, Czech Republic and Germany. The insect community identified on J. cyanoides and differences in its composition on robust populations of plant in contrast to sparsely populated periphery of its range are discussed. Altogether 78 species belonging to six orders (Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera and Neuroptera) were recorded. Larvae of the pyralid moth (genus Dioryctria), tephritid fruit flies (Acanthiophilus helianthi), and several polyphagous Heteroptera species are identified as main taxa feeding on inflorescences and seeds of J. cyanoides, with a potential to reduce the plant’s sexual reproduction. The impact of phytophagous insects is, however, considered only a secondary reason for the decline of the Czech populations of J. cyanoides on which a fairly low number of insect species were recorded in comparison with Ukraine and Germany; habitat loss and inbreeding effect are probably major negative factors. Several new host plant-insect and host-parasitoid associations are reported for insects on J. cyanoides.

Investigation of the midgut structure and ultrastructure in Cimex lectularius and Cimex pipistrelli (Hemiptera: Cimicidae)

Cimicidae are temporary ectoparasites, which means that they cannot obtain food continuously. Both Cimex species examined here, Cimex lectularius (Linnaeus 1758) and Cimex pipistrelli (Jenyns 1839), can feed on a non-natal host, C. lectularius from humans on bats, C. pipistrelli on humans, but never naturally. The midgut of C. lectularius and C. pipistrelli is composed of three distinct regions—the anterior midgut (AMG), which has a sack-like shape, the long tube-shaped middle midgut (MMG), and the posterior midgut (PMG). The different ultrastructures of the AMG, MMG, and PMG in both of the species examined suggest that these regions must fulfill different functions in the digestive system. Ultrastructural analysis showed that the AMG fulfills the role of storing food and synthesizing and secreting enzymes, while the MMG is the main organ for the synthesis of enzymes, secretion, and the storage of the reserve material. Additionally, both regions, the AMG and MMG, are involved in water absorption in the digestive system of both Cimex species. The PMG is the part of the midgut in which spherites accumulate. The results of our studies confirm the suggestion of former authors that the structure of the digestive tract of insects is not attributed solely to diet but to the basic adaptation of an ancestor.

Long-term changes of small mammal communities in heterogenous landscapes of Central Europe

Long-term dynamics of small mammal communities are perennial themes in population ecology. However, comprehensive studies on the effect of environmental factors on population dynamics are still rare. Here, we aimed to analyze long-term data on Central European communities of small mammals occurring in two habitats that greatly differed in their structure, successional stages, and forest management. We found a richer community structure in young spruce plantations compared to mature European beech forests. In young spruce plantations, Myodes glareolus and Apodemus flavicollis abundances increased and Sorex araneus abundances decreased during the study period as a result of forest growth and management. Community structure in mature beech forests did not change significantly during the study period. Apodemus flavicollis and Myodes glareolus showed 3- and 5-year population cycles, respectively, and their abundances were simultaneously positively correlated with relative abundance of masts. Weather also played a role, while the effect of snow cover was pronounced only in mountain areas where it negatively affected Microtus agrestis and Sorex araneus abundances, temperature positively and rainfall negatively influenced Myodes glareolus and Apodemus flavicollis abundances across both studied habitats. Our findings document that a complex of environmental factors significantly affects the structure and dynamics of small mammal communities in Central Europe, and both local biotic and abiotic factors should be considered in future studies.