In vitro comparison of two food choice methodologies for necrophagous species of the genus Thanatophilus (Coleoptera: Staphylinidae: Silphinae).

To accurately model the food webs, we need to acquire precise data on food ecology of the interacting species. This allows better understanding of the trophic interactions and for the necrophagous species this information could be used in medico-legal investigations. For this reason, we recently proposed standardized laboratory methodology to assess the foraging strategies based on parallel testing of 2 food items (meat, dead larvae) (Jakubec et al. 2021). The original methodology had 2 shortcomings. It was not suited for testing living larvae, which could prove predatory behavior of the species. The methodology was also based on parallel experimental design, where the food items are tested together, which could underestimate the maximum consumption of the tested subject for some items. To test if these concerns are valid, we improved original methodology allowing testing living larvae as well as a new sequential experimental setup, where consumption of each item is tested individually in a random order, thus theoretically giving an unbiased maximum consumption estimate. These methodologies were tested head-to-head on 3 forensically relevant species from the genus Thanatophilus (Thanatophilus micans (Fabricius 1794)(Fabricius 1794), Thanatophilus rugosus (Linnaeus, 1758), and Thanatophilus sinuatus(Fabricius, 1775)). The experiments have confirmed that all 3 species are almost strictly necrophagous, although they were capable of predation, despite the presence of preferred food (meat). The comparison also showed that the sequential design has indeed improved capability to quantify the maximal consumption of the given food item. Thus, we suggest following this methodology in future studies.

Resurrection of Oxythyrea abigailoides Miksic, 1978 (Coleoptera: Scarabaeidae: Cetoniinae) based on new morphological, morphometrical and molecular data.

Oxythyrea abigailoides Miksic, 1978, resurrected species, is removed from synonymy with Oxythyrea dulcis Reitter, 1899 and regarded as a separate species. The differential diagnosis of both species is presented on the basis of clearly different morphological, morphometric and molecular characters. Complete chresonomy to both species is given and known geographical distribution to date is summarized and mapped.

Emotions triggered by live arthropods shed light on spider phobia.

Spiders are mostly harmless, yet they often trigger high levels of both fear and disgust, and arachnophobia (the phobia of spiders) ranks among the most common specific animal phobias. To investigate this apparent paradox, we turned to the only close relatives of spiders that pose a real danger to humans: scorpions. We adopted a unique methodology in order to assess authentic emotions elicited by arthropods. Over 300 respondents were asked to rate live specimens of 62 arthropod species (including spiders, scorpions, cockroaches, and other insects) based on perceived fear, disgust, and beauty. We found that species' scores on all three scales depended on the higher taxon as well as on body size. Spiders, scorpions, and other arachnids scored the highest in fear and disgust, while beetles and crabs scored the highest in beauty. Moreover, all chelicerates were perceived as one cohesive group, distinct from other arthropods, such as insects or crabs. Based on these results, we hypothesize that the fear of spiders might be triggered by a generalized fear of chelicerates, with scorpions being the original stimulus that signals danger.

Standardized Laboratory Methodology for the Evaluation of Foraging Strategies in Necrophilous Beetles: A Case Study of Necrophila (Calosilpha) brunnicollis (Coleoptera: Silphidae).

Precise data regarding feeding habits of necrobiont species are a key element of food web and evolutionary ecology. They can also be used to assess the utility and value of those species for forensic entomology, where obligatory necrophagous species in particular are considered good bioindicators of postmortem or preappearance interval. However, the feeding habits of many species are known only from anecdotal field observations, often reduced to vaguely defined categories-predatory, necrophagous, or omnivorous. To address this issue, we designed a simple, in vitro behavioral experiment allowing the quantification of food preferences. Next, we applied it on Necrophila (Calosilpha) brunnicollis (Kraatz, 1877), which is a common carrion beetle of East Asia with unresolved food preferences. The results suggest that this species is preferentially necrophagous, thus valuable for forensic research. Importantly, however, our experimental design allowed us to reveal that it also readily feeds on larvae of Diptera, although they compose a minor proportion of its diet. This methodology can be applied to other species, and it could provide evidence for future decision making in forensic research.

Pachnoda iskuulka (Coleoptera: Scarabaeidae: Cetoniinae), a new species from Somaliland, including description of its mature larva.

Pachnoda iskuulka Král, Sommer Sípek, new species from the Sanaag region of north-eastern Somaliland is described. The new species is compared with the morphologically similar taxa Pachnoda abyssinica abyssinica Reiche, 1847, P. a. meriteti Di Gennaro, 2017, P. massajae Gestro, 1881, and P. werneri Beinhundner, 1992, all of which occur in the Horn of Africa. Relevant diagnostic characters (e.g., dorsal and ventral surface color pattern, male external genitalia) are illustrated. The third instar larva of the new species is described and biological notes are provided.

New species of Xiphoscelis Burmeister, 1842 (Coleoptera, Scarabaeidae, Cetoniinae) from arid regions of South Africa and Namibia.

Two new species of the southern African genus Xiphoscelis Burmeister, 1842 are recognised and described, X. braunsi sp. nov. from the Eastern and Western Cape Karoo (South Africa) and X. namibica sp. nov. from the Huns Mountains of southern Namibia and adjacent ranges in South Africa. These were previously overlooked and grouped together with X. schuckardi Burmeister, 1842, but further material and more in-depth analyses have now revealed their clear separation on the basis of key diagnostic features, including clypeal structure, metatibial spur development and aedeagal shape. The densely and coarsely costate elytral structure and the black to brown colour of these species are symplesiomorphies shared with a number of the most primitive genera among the African Cetoniinae. However, these characters also reflect the convergent adaptation to hot and arid conditions they share with several other species occurring in this region. Phylogenetic relationships of the genus with other Cetoniinae are explored using the larval characters highlighted in the description of the 3rd instar larva of X. braunsi sp. nov. The extraordinary hypertrophy observed in the male metatibial spur of species in this genus, and particularly in X. schuckardi, appears to represent a defence mechanism against potential predators on the ground, apart from playing a role during mating.

Lethrus (Lethrus) thracicus (Coleoptera: Geotrupidae), a new species from the European part of Turkey.

Lethrus (Lethrus) thracicus Král Hillert, new species, from Thrace, north-western Turkey is described. The new species is compared with the morphologically similar species L. (L.) apterus (Laxmann, 1770), L. (L.) ares Král, Rejsek Schneider, 2001 and L. (L.) schneideri Král Hillert, 2013. Relevant diagnostic male characters (shape of mandibles, ventral mandible processes, pronotum and parameres) are illustrated. The name Lethrus subaeneus Fairmaire, 1866 is considered a nomen nudum.

Immature stages of three species of Inca LePeletier Serville, 1828 (Coleoptera: Scarabaeidae: Cetoniinae) and morphology of phytophagous scarab beetle pupa.

Immature stages of I. pulverulenta (Olivier, 1789) (Coleoptera: Scarabaeidae: Cetoniinae: Incini) are described for the first time, and those of I. bonpalandi (Gyllenhal, 1827) and I. clathrata sommeri Westwood, 1845 are redescribed. A key to the third instar of Incini is presented, notes on Inca larval characters in the phylogenetic context of Cetoniinae and on scarab pupal morphology are also given.

Immature stages of the genus Oxythyrea (Coleoptera: Scarabaeidae: Cetoniinae) with a key to third instar larvae, and notes on the biology of the genus.

The genus Oxythyrea Mulsant, 1842 comprises ten currently recognized species. So far, larval descriptions for only three species are available. Here, we present comprehensive descriptions of third instar larval morphology for the six previously unstudied Oxythyrea species along with detailed re-descriptions of larval morphology for the previously described species. All descriptions are supplemented by photographs and line art drawings of the morphological structures. This work also contains a key to third instar larvae of the genus Oxythyrea as well as observations from collecting sites with focus on biological characteristics of all covered species including pictures of biotopes and host plants for adults. In addition, notes on the breeding circumstances for species are provided.

Molecular and Morphological Diversity of the Oxymonad Genera Monocercomonoides and Blattamonas gen. nov.

Oxymonads are a group of flagellates living as gut symbionts of insects or vertebrates. They have several unique features, one of them being the absence of mitochondria. Diversity of this group is seriously understudied, which is particularly true for small species from the family Polymastigidae. We isolated 34 strains of oxymonads with Polymastigidae-like morphology from 24 host species and unused cesspits and sequenced the SSU rRNA gene. Our strains formed two clades in the phylogenetic tree with Streblomastix strix branching between them. This topology was also supported by a three-gene phylogenetic analysis. Despite considerable genetic differences between the clades, light and electron microscopy revealed only subtle differences. The larger clade is considered genus Monocercomonoides and the isolates belonging here were classified into three new species (including the first potentially free-living species), two previously described species, and three unclassified lineages. The smaller clade, here described as Blattamonas gen. nov., consists of three newly described species. Concomitantly with the description of Blattamonas, we elevate the Monocercomonoides subgenus Brachymonas to the genus level. Our study shows that, despite their conserved morphology, the molecular diversity of Polymastigidae-like oxymonads is broad and represents a substantial part of the diversity of oxymonads.

Hidden complexity in the ontogeny of sexual size dimorphism in male-larger beetles.

Sexual size dimorphism (SSD) is widespread among animals, but its developmental mechanisms are not fully undestood. We investigated the proximate causes of SSD in three male-larger and one monomorphic scarab beetles using detailed monitoring of growth in individual instars. Apart from the finding that SSD in all three male-larger species started to develop already in the first larval instar, we generally found a high variability in SSD formation among the species as well as among instars. Overall, sexual differences in developmental time, average growth rate, as well as in the shape of the growth trajectory seem to be the mechanisms responsible for SSD ontogeny in scarab beetles. In the third instar, when the larvae attain most of their mass, the males had a similar or even lower instantaneous growth rate than females and SSD largely developed as a consequence of a longer period of rapid growth in males even in cases when the sexes did not differ in the total duration of this instar. Our results demonstrate that a detailed approach, examining not only the average growth rate and developmental time, but also the shape of the growth trajectory, is necessary to elucidate the complex development of SSD.

Phylogeography and DNA-based species delimitation provide insight into the taxonomy of the polymorphic rose chafer Protaetia (Potosia) cuprea species complex (Coleoptera: Scarabaeidae: Cetoniinae) in the Western Palearctic.

The development of modern methods of species delimitation, unified under the "integrated taxonomy" approach, allows a critical examination and re-evaluation of complex taxonomic groups. The rose chafer Protaetia (Potosia) cuprea is a highly polymorphic species group with a large distribution range. Despite its overall commonness, its taxonomy is unclear and subject to conflicting hypotheses, most of which largely fail to account for its evolutionary history. Based on the sequences of two mitochondrial markers from 65 individuals collected across the species range, and a detailed analysis of morphological characters including a geometric morphometry approach, we infer the evolutionary history and phylogeography of the P. cuprea species complex. Our results demonstrate the existence of three separate lineages in the Western Palearctic region, presumably with a species status. However, these lineages are in conflict with current taxonomic concepts. None of the 29 analyzed morphological characters commonly used in the taxonomy of this group proved to be unambiguously species- or subspecies- specific. The geometric morphometry analysis reveals a large overlap in the shape of the analyzed structures (pronotum, meso-metaventral projection, elytra and aedeagus), failing to identify either the genetically detected clades or the classical species entities. Our results question the monophyly of P. cuprea in regard to P. cuprina, as well as the species status of P. metallica. On the other hand, we found support for the species status of the Sicilian P. hypocrita. Collectively, our findings provide a new and original insight into the taxonomy and phylogeny of the P. cuprea species complex. At the same time, the results represent the first attempt to elucidate the phylogeography of these polymorphic beetles.

Immature stages of giants: morphology and growth characteristics of Goliathus Lamarck, 1801 larvae indicate a predatory way of life (Coleoptera, Scarabaeidae, Cetoniinae).

The third larval instar of Goliathus goliatus (Drury, 1770), Goliathus orientalis Moser, 1909 and Goliathus albosignatus Boheman, 1857 are described and illustrated for the first time and compared with the immature stages of other Cetoniinae. Larval development of Goliathus goliatus is investigated under laboratory conditions, with particular emphasis on food requirements. These results support the obligatory requirement of proteins in the larval diet. The association between larval morphological traits (e. g., the shape of the mandibles and pretarsus, presence of well-developed stemmata) and larval biology is discussed. Based on observations and the data from captive breeds it is concluded that a possible shift from pure saprophagy to an obligatory predaceous way of larval life occurred within the larvae of this genus, which may explain why these beetles achieve such an enormous size.

Ontogeny of sexual size dimorphism in the hornless rose chafer Pachnoda marginata (Coleoptera: Scarabaeidae: Cetoniinae).

Beetles of the subfamily Cetoniinae are distinct and well-known, yet their larval ontogeny, sexual size dimorphism and development remain unknown in most species. This group contains many species with large males with prominent secondary sexual structures, such as cephalic or pronotal horns and elongated forelimbs. The species studied here, Pachnoda marginata, belongs to those species without any obvious dimorphism, the males being almost indistinguishable from the females. In this paper we examine sexual dimorphism in body shape and size in this apparently 'non-dimorphic' species. We further investigate the larval development and proximate causes of sexual size dimorphism, in particular when and how the sexes diverge in their growth trajectories during ontogeny. We found that males are larger than females and that the sexes also differ in body shape - for example, males possess significantly longer forelimbs relative to body size than females. The male-biased sexual size dimorphism along with prolonged forelimbs suggests that sexual selection for larger males may not be limited merely to horned species of rose chafers. The dimorphism in size in P. marginata arises during the second larval instar and basically remains unchanged till maturity. In both sexes the maximum body mass as well as developmental time of particular larval instars were strongly correlated, but time spent in the pupal chamber was not related to previous growth and final body size. The correlation between developmental time and adult size was negative, which may be a reflection of differences in resource allocation or utilisation for growth and development among individuals.

A molecular phylogeny of rose chafers (Coleoptera: Scarabaeidae: Cetoniinae) reveals a complex and concerted morphological evolution related to their flight mode.

Rose chafers (Cetoniinae) are a large group of flower visitors within the pleurostict Scarabaeidae that are characterized by their distinctive flight mode with nearly closed forewings. Despite their popularity, this is the first study to use molecular data to infer their phylogenetic relationships. We used partial gene sequences for 28S rRNA, cytochrome oxidase I (cox1) and 16S rRNA (rrnL) for 299 species, representing most recognized subfamilies of Scarabaeidae, including 125 species of Cetoniinae. Combined analyses using maximum parsimony, maximum likelihood and Bayesian inferences recovered Cetoniinae as monophyletic in all analyses, with the sister clade composed of Rutelinae and Dynastinae. Rutelinae was always recovered as paraphyletic with respect to Dynastinae. Trichiini sensu lato (s.l.) was recovered as a polyphyletic clade, while Cetoniini s.l. was recovered as paraphyletic. The inferred topologies were also supported by site bootstrapping of the ML trees. With the exception of Cremastochelini, most tribes of Cetoniinae were poly- or paraphyletic, indicating the critical need for a careful revision of rose chafer classification. Analysis of elytral base structure (including 11 scored characters) in the context of phylogeny, revealed a complex, concerted and rapid transformation of the single trait elements linked to a modified flight mode with closed elytra. This appears to be unlinked to the lateral sinuation of the elytra, which originated independently several times at later stages in the evolution of the group.

Description of adult and third instar larva of Trichostetha curlei sp. n. (Coleoptera, Scarabaeidae, Cetoniinae) from the Cape region of South Africa.

A new high altitude montane species of Trichostetha Burmeister, 1842 is described from the Elandsberg range of the Western Cape interior. This represents the 14(th) species of the genus and the first to be reported with a description of its larva. It is a significant addition to the growing number of species that exhibit no adult feeding behaviour and a short period of activity restricted to the onset of summer. Larvae dwell in rock crevices, feeding on decomposing plant matter. The genus Trichostetha is heterogeneous and the complex variability observed in some species, especially T. capensis (Linnaeus, 1767), requires the re-instatement of taxa that were recently synonymised. Thus, T. bicolor Péringuey, 1907 is here re-proposed as a separate species and T. capensis hottentotta (Gory & Percheron, 1833) as a separate subspecies. Conversely, T. alutacea Allard, 1994 is recognised as a dark variety of T. signata (Fabricius, 1775) and is, consequently, synonymised with this species.

Lethrus (Lethrus) schneideri sp. n. (Coleoptera, Geotrupidae) from Greece.

Lethrus (Lethrus) schneideri Král & Hillert, sp. n. from Thrace, Greece, is described. The new species is morphologically most similar and probably closely related to Lethrus (Lethrus) apterus (Laxmann, 1770) and Lethrus (Lethrus) ares Král, Rejsek & Schneider, 2001. Diagnostic characters (shape of mandibles, ventral mandible processes, pronotum and parameres) are illustrated. Character matrix for separation of males of the Lethrus species closely related to Lethrus schneideri Král & Hillert, sp. n. and geographic ranges for all species studied are mapped.

Integrative analysis of DNA phylogeography and morphology of the European rose chafer (Cetonia aurata) to infer species taxonomy and patterns of postglacial colonisation in Europe.

Integrative taxonomy has been proposed as a framework to unify new conceptual and methodological developments in quantitative assessment of trait variation used in species delimitation, but empirical studies in this young branch of systematics are rare. Here we use standard phylogenetic and parsimony network analyses on nuclear and mitochondrial DNA (Cox1, ITS1) of 230 individuals from 65 European sampling sites in order to deduce population structure of Cetonia beetles from geno- and haplotypes. Statistical measures of population differentiation are inferred on genealogical and geographical scales to test hypotheses about species limits and population history. By combining results of phylogenetic structure with features of morphology, including genital shape morphometrics and discrete external body characters, as well as with measures of population genetics, we attempt to integrate the results as a test of the validity of species limits, in particular of currently recognised subspecies. Despite high Cox1 divergence between some haplotype lineages, even some sympatric lineages (9%, e.g. N2 vs. N4), nDNA and morphology, as well as pattern of geographical and genealogical divergence measured by AMOVA analysis did not support the hypothesis of separate species. Highest divergence in nuclear markers was found among Italian populations of C. aurata pisana and C. a. sicula, and moderately high fixation indices along measurable morphological divergence suggest the correctness of their status as 'subspecies'. Divergence time estimates of the lineages suggest a glacial divergence in different refugia between the major haplogroups, while population differentiation in mtDNA among these was primarily attributable to restricted gene flow caused by geographic isolation.

Immature stages and ecology of two species of the South African genus Stripsipher Gory & Percheron, 1833 (Coleoptera, Scarabaeidae, Cetoniinae, Trichiini).

Based on the study of newly accessible type material, Stripsipher drakensbergi Ricchiardi, 1998, is demoted to a junior synonym of Stripsipher jansoni Péringuey, 1908. The genus Stripsipher Gory & Percheron, 1833, thus, currently includes 12 species, but for none of these are larval stages and/or pupae currently known. The immature stages of Stripsipher orientalis Ricchiardi, 2008 and Stripsipher jansoni are described here for the first time and updated observations on distribution and ecology of both species are provided. Morphological affinities of Stripsipher with other Trichiini larvae are presented and the main diagnostic differences discussed. The larvae of both species are very similar to those of other representatives of the tribe Trichiini, with key differences found on the epipharynx. Based on the morphology of larvae and adults, it is suggested that Stripsipher is a member of the clade composed of Valgini, Trichiini and Cryptodontini.

Head rotations in the play of Hanuman langurs (Semnopithecus entellus): description and analysis of function.

Although head rotations are frequent patterns in play behavior in many mammalian species and differ from head movements used in other contexts, they have not been quantitatively described and their function remains unclear. The head rotations occurring in the play behavior of free-ranging Hanuman langurs (Semnopithecus entellus) were described from videotaped sequences. The authors tested 2 possible hypotheses about their function. Either the head rotations serve to create unexpected situations and should therefore occur in both solitary and social play and also be very variable, or they serve as play signals and should therefore occur only in social play and be ritualized. If head rotations have both functions, they should be less variable in social play. The data revealed that head rotations were very variable and were present both in solitary and social play. Furthermore, there was no difference in the variability between the head rotations present in the 2 types of play. The results do not support the function of head rotations as play signals but, rather, suggest that head rotations may serve to create unexpected situations in play.