Boring life: early colony formation and growth in the endolithic bryozoan genus Penetrantia Silén, 1946.

As in most colonial and sessile marine invertebrates, bryozoan life history is characterized by asexual propagation of zooids for colonial growth and by sexual production of larvae for dispersal. However, comprehensive life histories, particularly in cryptic species such as endolithic (boring) bryozoans, remain poorly understood. The ctenostome family Penetrantiidae is widespread from temperate to tropical waters and often found in molluscan shells, offering an opportunity to study the boring lifestyle and its potential impact on bioerosion through growth and settlement experiments. Our research focused on Penetrantia clionoides from Guam in the Pacific Ocean, Penetrantia japonica from Japan, and a Penetrantia species from France in the Atlantic Ocean. We found distinct life histories and reproductive patterns potentially influenced by environmental factors such as temperature and food availability. The tropical P. clionoides displayed higher rates of larval production and growth compared to its temperate counterpart. For instance, the mean stolon extension was 335.2 µm/week in P. clionoides versus 232.1 µm/week in Penetrantia sp. Autozooid development took 13 days in P. clionoides and 31 days in Penetrantia sp. Anatomical features like apertural rims aided in species identification and in understanding larval settlement preferences, suggesting a tendency for philopatric settlement behavior. The bioerosional impact of penetrantiids remains little understood, but we generated first projections of bioerosion rates and a protocol for keeping Penetrantia under laboratory conditions, laying a foundation for further research in this field.

Boring systematics: A genome skimmed phylogeny of ctenostome bryozoans and their endolithic family Penetrantiidae with the description of one new species.

Ctenostomes are a group of gymnolaemate bryozoans with an uncalcified chitinous body wall having few external, skeletal characters. Hence, species identification is challenging and their systematics remain poorly understood, even more so when they exhibit an endolithic (boring) lifestyle. Currently, there are four Recent families of endolithic bryozoans that live inside mineralized substrates like mollusk shells. In particular, Penetrantiidae Silén, 1946 has received considerable attention and its systematic affinity to either cheilostomes or ctenostomes has been debated. Species delimitation of penetrantiids remains difficult, owing to a high degree of colonial and zooidal plasticity. Consequently, an additional molecular approach is essential to unravel the systematics of penetrantiids, their phylogenetic placement and their species diversity. We therefore sequenced the mitochondrial (mt) genomes and two nuclear markers of 27 ctenostome species including nine penetrantiids. Our phylogeny supports the Penetrantiidae as a monophyletic group placed as sister taxon to the remaining ctenostomes alongside paludicellids, arachnidioids and terebriporids. The boring family Terebriporidae d'Orbigny, 1847 were previously considered to be among vesicularioids, but our results suggest an arachnidioid affinity instead. Ctenostome paraphyly is supported by our data, as the cheilostomes nest within them. A Multiporata clade is also well supported, including the former victorelloid genus Sundanella. Altogether, this study provides new insights into ctenostome systematics, assists with species delimitation and contributes to our understanding of the bryozoan tree of life.

Morphology of ctenostome bryozoans: 7. Hislopia, Echinella and Timwoodiellina.

Ctenostome bryozoans are a small group of gymnolaemates comprising less than 400 recent species. They are paraphyletic and ctenostome-grade ancestors gave rise to Cheilostomata, the most dominant and speciose taxon of Bryozoa in the present day. Investigations into ctenostomes are important for reconstructing character evolution among Gymnolaemata. As a continuation of studies on a morphological series of ctenostome bryozoans, we herein investigate six species of hislopiids, a small clade of three genera occurring in freshwater habitats. The general morphology of all species is similar in having primarily uniserial chains of encrusting zooids, which are mostly oval to ellipsoid and have a flattened frontobasal axis. Hislopia prolixa and Echinella placoides often have more slender zooids with a higher frontobasal axis. Apertures of hislopiids are quadrangular, lined by a thickened cuticle. Apertural spines are present in various lengths in E. placoides, Hislopia lacustris and Hislopia corderoi. The remaining cuticle is rather thin except at lateral areas, close to the pore-plates which are prominent in hislopiids because of abundant special and limiting cells. All species except H. corderoi and Timwoodiellina natans have a prominent collar obstructing the vestibulum, whereas the latter two species instead have an 'external collar' as cuticular, outer folds projecting over the aperture. Hislopiid lophophores carry eight, or more commonly 12-18 tentacles. The digestive tract is distinguished by an often highly elongated esophagus and/or cardia, with the latter always having a prominent bulbous part in the form of a proventriculus-or gizzard in E. placoides. The caecum is extensive in all species. In Hislopia the intestine is characteristically two-chambered with a proximal and distal part before entering an anal tube of various length. The latter is present in all species except T. natans and terminates in mid-lophophoral area. Oocytes in E. placoides are large and macrolecithal indicating brooding and the production of lecithotrophic larvae. Hislopia species produce small, oligolecithal ones, which suggests zygote spawning and planktotrophy. In general, the morphology is similar among the different hislopiids with characters of the gut aiding in delineating the genera Echinella and Timwoodiellina.

Delayed increase in stone tool cutting-edge productivity at the Middle-Upper Paleolithic transition in southern Jordan.

Although the lithic cutting-edge productivity has long been recognized as a quantifiable aspect of prehistoric human technological evolution, there remains uncertainty how the productivity changed during the Middle-to-Upper Paleolithic transition. Here we present the cutting-edge productivity of eight lithic assemblages in the eastern Mediterranean region that represent a chrono-cultural sequence including the Late Middle Paleolithic, Initial Upper Paleolithic, the Early Upper Paleolithic, and the Epipaleolithic. The results show that a major increase in the cutting-edge productivity does not coincide with the conventional Middle-Upper Paleolithic boundary characterized by the increase in blades in the Initial Upper Paleolithic, but it occurs later in association with the development of bladelet technology in the Early Upper Paleolithic. Given increasing discussions on the complexity of Middle-Upper Paleolithic cultural changes, it may be fruitful to have a long-term perspective and employ consistent criteria for diachronic comparisons to make objective assessment of how cultural changes proceeded across conventional chrono-cultural boundaries.

Boring bryozoans: an investigation into the endolithic bryozoan family Penetrantiidae.

An endolithic lifestyle in mineralized substrates has evolved multiple times in various phyla including Bryozoa. The family Penetrantiidae includes one genus with ten extant and two fossil species. They predominantly colonize the shells of molluscs and establish colonies by chemical dissolution of calcium carbonate. Based on several morphological characters, they were described to be either cheilostome or ctenostome bryozoans. For more than 40 years, neither the characters of species identity and systematics nor the problem of their phylogeny was approached. Consequently, the aim of this study is to reevaluate species identities and the systematic position of the genus Penetrantia by analyzing at least six different species from eight regions with the aid of modern methods such as confocal laser scanning microscopy and 3D-reconstruction techniques. This study demonstrates that the musculature associated with the operculum and brood chamber shows significant differences from the cheilostome counterparts and seems to have evolved independently. Together with the presence of other ctenostome-like features such as true polymorphic stolons and uncalcified body wall, this finding supports a ctenostome affinity. Operculum morphology reveals many new species-specific characters, which, together with information about gonozooid morphology, tentacle number, and zooid size ranges, will enhance species identification. It also revealed a probable new species in Japan as well as potential cryptic species in France and New Zealand. In addition, this study increases the known distribution range of the family and its substrate diversity. Altogether, the new information collated here provides the basis for future work on a neglected taxon. Supplementary Information: The online version contains supplementary material available at 10.1007/s13127-023-00612-z.

Phylogenomics reveals deep relationships and diversification within phylactolaemate bryozoans.

Bryozoans are mostly sessile colonial invertebrates that inhabit all kinds of aquatic ecosystems. Extant bryozoan species fall into two clades with one of them, Phylactolaemata, being the only exclusively freshwater clade. Phylogenetic relationships within the class Phylactolaemata have long been controversial owing to their limited distinguishable characteristics that reflect evolutionary relationships. Here, we present the first phylogenomic analysis of Phylactolaemata using transcriptomic data combined with dense taxon sampling of six families to better resolve the interrelationships and to estimate divergence time. Using maximum-likelihood and Bayesian inference approaches, we recovered a robust phylogeny for Phylactolaemata in which the interfamilial relationships are fully resolved. We show Stephanellidae is the sister taxon of all other phylactolaemates and confirm that Lophopodidae represents the second offshoot within the phylactolaemate tree. Plumatella fruticosa clearly falls outside Plumatellidae as previous investigations have suggested, and instead clusters with Pectinatellidae and Cristatellidae as the sister taxon of Fredericellidae. Our results demonstrate that cryptic speciation is very likely in F. sultana and in two species of Plumatella (P. repens and P. casmiana). Divergence time estimates show that Phylactolaemata appeared at the end of the Ediacaran and started to diverge in the Silurian, although confidence intervals were large for most nodes. The radiation of most extant phylactolaemate families occurred mainly in the Palaeogene and Neogene highlighting post-extinction diversification.

Paleozoic origins of cheilostome bryozoans and their parental care inferred by a new genome-skimmed phylogeny.

Phylogenetic relationships and the timing of evolutionary events are essential for understanding evolution on longer time scales. Cheilostome bryozoans are a group of ubiquitous, species-rich, marine colonial organisms with an excellent fossil record but lack phylogenetic relationships inferred from molecular data. We present genome-skimmed data for 395 cheilostomes and combine these with 315 published sequences to infer relationships and the timing of key events among c. 500 cheilostome species. We find that named cheilostome genera and species are phylogenetically coherent, rendering fossil or contemporary specimens readily delimited using only skeletal morphology. Our phylogeny shows that parental care in the form of brooding evolved several times independently but was never lost in cheilostomes. Our fossil calibration, robust to varied assumptions, indicates that the cheilostome lineage and parental care therein could have Paleozoic origins, much older than the first known fossil record of cheilostomes in the Late Jurassic.

New Species of Lower-Shelf to Upper-Slope Ctenostome Bryozoans from Pacific Japan, with a Family Range Extension.

Ctenostome bryozoans were collected from depths of 150-300 m in Suruga Bay, the Kumano Sea, and off Sendai Bay on the Pacific coast of Japan. Among these samples were five new species, three of which were epibiotic on other animals. Alcyonidium clavum n. sp., found encrusting cirri of the stalked crinoid Metacrinus rotundus, was previously reported in Japan as Alcyonidium mamillatum, but differs from the latter in lacking rings on the peristome. Triticella parvacrista n. sp. and Triticella cunabula n. sp. were epibiotic on the isopod Bathynomus doederleini and the pycnogonid Ascorhynchus japonicum, respectively. This is the first record of the genus Triticella from Japanese waters. These species differ from previously described species in autozooidal morphology, particularly the dimensions of the autozooidal dilatation, in the presence or absence of the frenaculum, and in the septate junction between the dilatation and pedicel. Bockiella arcatumida n. sp., found as erect colonies on pebbles and hydroids collected off Sendai Bay, differs from previously known species in the size and arrangement of the kenozoids in the branch. Metalcyonidium morum n. sp., collected southwest of the Izu Peninsula, occurred as small colonies having a peduncle of short kenozooids. This is the first record from the northwestern Pacific for both the genus Metalcyonidium and the family Clavoporidae. Metalcyonidium morum n. sp differs from Metalcyonidium gautieri, the only other species known in the genus, in having the capitulum divided into two parts, and in the length of the kenozooids comprising the peduncle.

Developmental Process of a Heterozooid: Avicularium Formation in a Bryozoan, Bugulina californica.

In bryozoans (phylum Bryozoa), representative colonial animals mostly found in marine environments, some species possess different types of individuals (heterozooids) specialized in different functions such as defense or structural support for their colonies. Among them, the best-known heterozooids are the avicularia, known to function as defenders. The differentiation processes of heterozooids, including avicularia, should be important keys to understand the evolutionary significance of bryozoans. However, the developmental process of avicularium formation remains to be fully understood. In this study, therefore, in order to understand the detailed developmental process and timing of avicularium formation, extensive observations were carried out in a bryozoan species, Bugulina californica (Cheilostomata, Bugulidae), that possesses adventitious avicularia, by performing stereomicroscopy on live materials, in addition to scanning electron microscopy and histological observations. The whole process can be divided into seven stages based on developmental events. Especially notably, at the earlier stages, there are three major budding events that produce proliferating cell masses corresponding to primordial tissues: (1) budding of the peduncle cushion at the outer margin of the distal part of a young autozooid, (2) budding of the head-part primordium from the peduncle cushion, and (3) budding of the polypide inside the head part. Experimental control of temperature showed that 20°C would be the best to maintain B. californica colonies.

Morphology of Stephanella hina (Bryozoa, Phylactolaemata): common phylactolaemate and unexpected, unique characters.

Stephanella hina is a little studied freshwater bryozoan belonging to Phylactolaemata. It is currently the only representative of the family Stephanellidae, which in most reconstructions is early branching, sometimes even sister group to the remaining phylactolaemate families. The morphological and histological details of this species are entirely unknown. Consequently, the main aim of this study was to conduct a detailed morphological analysis of S. hina using histological serial sections, 3D reconstruction, immunocytochemical staining and confocal laser scanning microscopy techniques. The general morphology is reminiscent of other phylactolaemates; however, there are several, probably apomorphic, details characteristic of S. hina. The most evident difference lies in the lophophoral base, where the ganglionic horns/extensions do not follow the traverse of the lophophoral arms but bend medially inwards towards the mouth opening. Likewise, the paired forked canal does not fuse medially in the lophophoral concavity as found in all other phylactolaemates. Additional smaller differences are also found in the neuro-muscular system: the rooting of the tentacle muscle is less complex than in other phylactolaemates, the funiculus lacks longitudinal muscles, the caecum has smooth muscle fibres, latero-abfrontal tentacle nerves are not detected and the medio-frontal nerves mostly emerge directly from the circum-oral nerve ring. In the apertural area, several neurite bundles extend into the vestibular wall and probably innervate neurosecretory cells surrounding the orifice. These morphological characteristics support the distinct placement of this species in a separate family. Whether these characteristics are apomorphic or possibly shared with other phylactolaemates will require the study of the early branching Lophopodidae, which remains one of the least studied taxa to date.

A genome-skimmed phylogeny of a widespread bryozoan family, Adeonidae.

BACKGROUND: Understanding the phylogenetic relationships among species is one of the main goals of systematic biology. Simultaneously, credible phylogenetic hypotheses are often the first requirement for unveiling the evolutionary history of traits and for modelling macroevolutionary processes. However, many non-model taxa have not yet been sequenced to an extent such that statistically well-supported molecular phylogenies can be constructed for these purposes. Here, we use a genome-skimming approach to extract sequence information for 15 mitochondrial and 2 ribosomal operon genes from the cheilostome bryozoan family, Adeonidae, Busk, 1884, whose current systematics is based purely on morphological traits. The members of the Adeonidae are, like all cheilostome bryozoans, benthic, colonial, marine organisms. Adeonids are also geographically widely-distributed, often locally common, and are sometimes important habitat-builders. RESULTS: We successfully genome-skimmed 35 adeonid colonies representing 6 genera (Adeona, Adeonellopsis, Bracebridgia, Adeonella, Laminopora and Cucullipora). We also contributed 16 new, circularised mitochondrial genomes to the eight previously published for cheilostome bryozoans. Using the aforementioned mitochondrial and ribosomal genes, we inferred the relationships among these 35 samples. Contrary to some previous suggestions, the Adeonidae is a robustly supported monophyletic clade. However, the genera Adeonella and Laminopora are in need of revision: Adeonella is polyphyletic and Laminopora paraphyletically forms a clade with some Adeonella species. Additionally, we assign a sequence clustering identity using cox1 barcoding region of 99% at the species and 83% at the genus level. CONCLUSIONS: We provide sequence data, obtained via genome-skimming, that greatly increases the resolution of the phylogenetic relationships within the adeonids. We present a highly-supported topology based on 17 genes and substantially increase availability of circularised cheilostome mitochondrial genomes, and highlight how we can extend our pipeline to other bryozoans.

Lithic technology, chronology, and marine shells from Wadi Aghar, southern Jordan, and Initial Upper Paleolithic behaviors in the southern inland Levant.

The Initial Upper Paleolithic (IUP) temporally overlaps with the range expansion of Homo sapiens populations in various parts of Eurasia and is often considered a key archaeological phase for investigating behavioral changes from the Middle Paleolithic. This paper reports upon new data from IUP occupations at Wadi Aghar, a rock shelter site in the southern Levant. In combining the results of radiometric dates and lithic analyses, we clarify the chronological and cultural position of Wadi Aghar assemblages in the Levantine IUP. As for the records about mobility, on-site activities, and resource procurement behaviors, we present analyses of lithic use-wear, tool-type composition, soil micromorphology, and marine shells. The lithic analyses and the optically stimulated luminescence (and subsidiary radiocarbon) dating of the Wadi Aghar materials suggest their chronocultural position in the IUP (45-40 ka for Layers C-D1; 39-36 ka for Layer B; possibly 50 ka for Layer D2), providing the southernmost location for the IUP in Eurasia. In the Levant, Wadi Aghar represents one of the few IUP sites in the inland areas. The results also indicate that the timing and technological sequences from the IUP to the following bladelet industries differed between the inland and coastal zones, likely reflecting geographically variable adaptive behaviors and/or cultural transmissions. One of the behavioral characteristics of IUP foragers at Wadi Aghar is the procurement of remote resources, represented by the transportation of marine shells from the Red Sea: Canarium fusiforme and Canarium cf. mutabile. Whether it was a direct procurement with increased mobility or a result of intergroup exchanges, it was not part of behavioral repertoires during the late MP in the same area. This can be understood as the expansion of resource procurement range, functioning as additional buffers from risk in the semiarid environments in the inland Levant.

The life of the freshwater bryozoan Stephanella hina (Bryozoa, Phylactolaemata)-a crucial key to elucidating bryozoan evolution.

BACKGROUND: Phylactolaemata is the earliest branch and the sister group to all extant bryozoans. It is considered a small relict group that, perhaps due to the invasion of freshwater, has retained ancestral features. Reconstruction of the ground pattern of Phylactolaemata is thus essential for reconstructing the ground pattern of all Bryozoa, and for inferring phylogenetic relationships to possible sister taxa. It is well known that Stephanella hina, the sole member of the family Stephanelllidae, is probably one of the earliest offshoots among the Phylactolaemata and shows some morphological peculiarities. However, key aspects of its biology are largely unknown. The aim of the present study was to analyze live specimens of this species, in order to both document its behavior and describe its colony morphology. RESULTS: The colony morphology of Stephanella hina consists of zooidal arrangements with lateral budding sites reminiscent of other bryozoan taxa, i.e., Steno- and Gymnolaemata. Zooids protrude vertically from the substrate and are covered in a non-rigid jelly-like ectocyst. The latter is a transparent, sticky hull that for the most part shows no distinct connection to the endocyst. Interestingly, individual zooids can be readily separated from the rest of the colony. The loose tube-like ectocyst can be removed from the animals that produces individuals that are unable to retract their lophophore, but merely shorten their trunk by contraction of the retractor muscles. CONCLUSIONS: These observations indicate that S. hina is unique among Phylactolaemata and support the notion that bryozoans evolved from worm-like ancestors. In addition, we raise several arguments for its placement into a separate family, Stephanellidae, rather than among the Plumatellidae, as previously suggested.

Description and molecular phylogeny of a new species of Phoronis (Phoronida) from Japan, with a redescription of topotypes of P. ijimai Oka, 1897.

We describe Phoronis emigi sp. n. as the eighth member of the genus based on specimens collected from a sandy bottom at 33.2 m depth in Tomioka Bay, Amakusa, Japan. The new species is morphologically similar to P. psammophila Cori, 1889, but can be distinguished from the latter by the number of longitudinal muscle bundles in the body wall (56-72 vs. 25-50 in P. psammophila) and the position of the nephridiopores (situated level with the anus vs. lower than the anus in P. psammophila). Using sequences of the nuclear 18S and 28S rRNA genes and the mitochondrial cytochrome c oxidase subunit I (COI) gene, we inferred the relationship of P. emigi to other phoronids by the maximum likelihood method and Bayesian analysis. The analyses showed that P. emigi is closely related to P. hippocrepia Wright, 1856 and P. psammophila Cori, 1889. We describe the morphology of the topotypes and additional material for P. ijimai Oka, 1897. Neither our morphological observations of P. ijimai, nor the phylogenetic analyses based on 18S and COI sequences, contradicts that P. vancouverensis Pixell, 1912 is conspecific with P. ijimai, a synonymy that has long been disputed.

Revision of the genus Buchneria (Bryozoa, Cheilostomata) from Japan.

Buchneria dofleini (Buchner, 1924), type species of Buchneria Harmer, 1957,was first described from material collected in 1904-1905 from Sagami Bay, Japan, but the type specimens had not been reexamined since the original description. In this study, I examined specimens of Buchneria from historical collections and material recently collected near Akkeshi, Hokkaido, Japan. Three Buchneria species were detected, two from Sagami Bay that Ortmann (1890) had placed in Escharoides, and one from Akkeshi that Androsova (1958) had described as Porella variabilis. I concluded that Buchneria dofleini is a junior synonym of Escharoides teres Ortmann, 1890; selected a lectotype for Escharoides teres among Ortmann's syntypes; and established the new combination Buchneria teres (Ortmann, 1890), which becomes the type species of Buchneria. I also established the new combination Buchneria rhomboidalis (Ortmann, 1890) and selected a lectotype among Ortmann's syntypes. Porella variabilis is transferred to Buchneria establishing the new combination Buchneria variabilis (Androsova, 1958). Here the three new combinations are redescribed and a key to the Japanese Buchneria species is provided. Finally, I transferred Buchneria to Bryocryptellidae on the basis of ovicell and orifice morphology. Therefore, Buchneria now includes a total of three species; Buchneria sinuata Harmer, 1957, a species from Indonesia that has hitherto been placed in this genus, is almost certainly not congeneric with other Buchneria. As far as is now known, Buchneria is endemic to northern Japan and the northern Sea of Japan.

Are plumatellid statoblasts in freshwater bryozoans phylogenetically informative?

Morphological characters of statoblasts (including floatoblasts and sessoblasts) in freshwater bryozoans have been important in phylactolaemate systematics and identification in that older phylogenetic hypotheses relied heavily on statoblast morphology. To assess the reliability of statoblast characters in drawing conclusions about phylogeny, we examined the phylogenetic distribution of metric and proportional floatoblast characters, floatoblast symmetry, and floatoblast and sessoblast microsculpture in Plumatellidae, the largest family of phylactolaemates, in the context of molecular phylogenetic reconstructions based on nucleotide sequences of parts of the mitochondrial 12S rRNA, 16S rRNA, and cytochrome b (cytb) genes. Adding cytb sequences to a previous phylogeny based on 12S and 16S increased support only for a node including Plumatella vaihiriae in the P. repens clade. Characters of gross floatoblast morphology were generally not phylogenetically informative individually, but collectively discriminated among members of three relatively well-supported clades that were analyzed as pre-defined groups in a discriminant analysis. Two characters of floatoblast microsculpture (reticulation and hypertubercles) were restricted to particular clades; other characters (e.g., villi) were clearly convergent. In nine of 11 cases, fenestral microsculpture was identical or partly correlated between the floatoblast and sessoblast of a species. Overall, our results indicate that statoblast morphology is not highly phylogenetically constrained.

Freshwater bryozoa of Tonle Sap, Cambodia.

We identified a collection of freshwater bryozoans from Tonle Sap (meaning Tonle Lake), Cambodia, a body of water fed by the Mekong River and characterized by extreme fluctuations in water level between the wet and dry seasons. The collection also included specimens from the moat of Angkor Wat, located at the north end of the lake. We found four phylactolaemate species (Plumatella bombayensis, Plumatella casmiana, Plumatella vorstmani, Hyalinella lendenfeldi) and one ctenostome species (Hislopia cambodgiensis) from the lake, and only a single, additional phylactolaemate species (Plumatella javanica) from the moat. We provide brief descriptions of these species, photographs of colonies for some, and photomicrographs by light and scanning electron microscopy (SEM) of statoblasts. None of the species encountered in this study is endemic to Cambodia, and the wide distributions of the species are possibly related to the dispersability of floatoblasts by birds. We briefly discuss some of the taxonomic problems surrounding Hislopia cambodgiensis.

Honda humanoid robots development.

Honda has been doing research on robotics since 1986 with a focus upon bipedal walking technology. The research started with straight and static walking of the first prototype two-legged robot. Now, the continuous transition from walking in a straight line to making a turn has been achieved with the latest humanoid robot ASIMO. ASIMO is the most advanced robot of Honda so far in the mechanism and the control system. ASIMO's configuration allows it to operate freely in the human living space. It could be of practical help to humans with its ability of five-finger arms as well as its walking function. The target of further development of ASIMO is to develop a robot to improve life in human society. Much development work will be continued both mechanically and electronically, staying true to Honda's 'challenging spirit'.