Early Cretaceous record of microboring organisms in skeletons of growing corals

Ko?odziej, B., Golubic, S., Bucur, I.I., Radtke, G. & Tribollet, A. 2011: Early Cretaceous record of microboring organisms in skeletons of growing corals. Lethaia, Vol. 45, pp. 34–45. A spectacularly preserved assemblage of microbial euendoliths, penetrating into skeletons of growing scleractinian corals, has been recognized in Early Aptian (Early Cretaceous) reef limestone of the Rar?u Mountains (East Carpathians, NE Romania). Microboring euendolithic filaments were found in five coral colonies of the suborder Microsolenina. They remained in part well‐preserved, often impregnated with iron oxides, which made them visible even in strongly recrystallized parts of coral skeletons. Filaments of a wide range of sizes (2–40 μm in diameter) were concentrated within medium parts of coral septa, oriented along the septa in the direction of the coral growth. The larger filaments were tubular, occurring in bundles and branched into finer, often tapering branches. Their behaviour and organization were quite similar to the modern euendolithic siphonalean chlorophyte Ostreobium. Filament diameters exceeded those reported for the modern species, but covered a similarly wide size range. Narrower frequently branching filaments, 4–8 μm in diameter, resemble distal branching patterns of modern Ostreobium quekettii. Some very thin filaments (ca. 1–2 μm) observed within skeleton or inside the large tubular filaments, sometimes associated with globular swellings, may represent euendolithic fungi. The recrystallization of coral skeleton had limited effect on preservation of euendoliths due to their impregnation with iron oxides; microbial euendoliths were subjected to different taphonomic changes. □Chlorophytes, Early Cretaceous, fungi, microbial euendoliths, Romania, scleractinian corals.     

Biomass of the cryptoendolithic microbiota from the Antarctic desert.

Extractable lipid phosphate was used to determine the biomass of the cryptoendolithic microbiota that colonizes sandstone rocks in the Ross Desert region of Antarctica. The mean amount of lipid phosphate was 0.053 micromole/cm2 (n = 9), which equals 2.54 g of C per m2 (range, 1.92 to 3.26 g of C per m2) of biomass in the biotic zone of these rocks. The turnover of phospholipids was comparable to that of temperate sediments and soils (t1/2, 6 to 9 days) at 0 degrees C and a light intensity of 305 micromoles of photons per m2 per s, indicating that this was a good method to measure viable biomass. The biomass was 0.3 to 9.6% of the total carbon content of the biotic zone and was about 2 orders of magnitude smaller than the epilithic lichen dry weight at a location some 7 degrees north in latitude. The cryptoendolithic microbiota had a uniform density throughout the biotic zone under the rock surface. The results indicate that the cryptoendolithic microbial biomass is small but viable in this unique, extreme ecosystem.     

Pyritized preservation of chancelloriids from the Cambrian Stage 3 of South China and implications for biomineralization

The enigmatic Cambrian animal chancelloriids were discovered in a wide range of taphonomic settings; however, preservation of biomineralized sclerite microstructure was solely known from secondarily phosphatized skeletal remains. Here, we investigate a uniquely pyritized chancelloriid from the lower Cambrian Guojiaba Formation in southern Shaanxi Province, China, using a combination of advanced analytic techniques. Results of the energy dispersive spectroscopy (EDS), X-Ray Fluorescence (XRF), and Raman spectrum show that the sclerites and scleritomes are preserved as pyritized internal moulds with a calcitic outer layer. The outer layer enveloping the internal moulds likely represents the recrystallized counterpart of the original biomineralized sclerite wall. Distinctive fibrous microstructures are discovered in the sclerites, which echo the features seen in the phosphatized fossils of chancelloriids. The typical microstructure, along with the recrystallized calcite, corroborate the interpretation that chancelloriid sclerites were originally constructed by fibrous aragonite. The stability of the microstructure and mineral composition in both carbonate and siliciclastic backgrounds indicate that chancelloriids were adapted to exploit aragonitic fibres to build their skeletons regardless of the change of their living environments.     

Biomass of the cryptoendolithic microbiota from the Antarctic desert

Extractable lipid phosphate was used to determine the biomass of the cryptoendolithic microbiota that colonizes sandstone rocks in the Ross Desert region of Antarctica. The mean amount of lipid phosphate was 0.053 micromole/cm2 (n = 9), which equals 2.54 g of C per m2 (range, 1.92 to 3.26 g of C per m2) of biomass in the biotic zone of these rocks. The turnover of phospholipids was comparable to that of temperate sediments and soils (t1/2, 6 to 9 days) at 0 degrees C and a light intensity of 305 micromoles of photons per m2 per s, indicating that this was a good method to measure viable biomass. The biomass was 0.3 to 9.6% of the total carbon content of the biotic zone and was about 2 orders of magnitude smaller than the epilithic lichen dry weight at a location some 7 degrees north in latitude. The cryptoendolithic microbiota had a uniform density throughout the biotic zone under the rock surface. The results indicate that the cryptoendolithic microbial biomass is small but viable in this unique, extreme ecosystem.     

我国寒武系底部几种磷酸盐化保存的蓝菌类化石

本文系统描述了产于我国寒武系底部磷矿层中的微型蓝菌类化石4属4种,其中Girvanella sp.和Spirellus columnaris产于新疆阿克苏、乌什玉尔吐斯组和云南会泽、安宁朱家箐组中谊村段,Endoconchia lata产于四川峨眉灯影组麦地坪段,Cambricodium capilloides和一块属种名未定的化石产于陕西宁强灯影组宽川铺段,从而丰富了我国寒武系底部磷块岩层中的蓝菌类化石。本文还讨论了上述微型蓝菌类化石有直接磷酸盐化保存和钻孔磷酸盐铸模保存的两种保存方式。     

Extensive metazoan reefs from the Ediacaran Nama Group,Namibia: the rise of benthic suspension feeding

We describe new, ecologically complex reef types from the Ediacaran Nama Group, Namibia, dated at ~548 million years ago (Ma), where the earliest known skeletal metazoans, Cloudina riemkeae and Namacalathus, formed extensive reefs up to 20 m in height and width. C. riemkeae formed densely aggregating assemblages associated with microbialite and thrombolite, each from 30 to 100 mm high, which successively colonised former generations to create stacked laminar or columnar reef frameworks. C. riemkeae individuals show budding, multiple, radiating attachment sites and cementation between individuals. Isolated Namacalathus either intergrew with C. riemkeae or formed dense, monospecific aggregations succeeding C. riemkeae frameworks, providing a potential example of environmentally mediated ecological succession. Cloudina and Namacalathus also grow cryptically, either as pendent aggregations from laminar crypt ceilings in microbial framework reefs or as clusters associated with thrombolite attached to neptunian dyke walls. These reefs are notable for their size, exceeding that of the succeeding Lower Cambrian archaeocyath–microbial communities. The repeated colonisation shown by C. riemkeae of former assemblages implies philopatric larval aggregation to colonise limited favourable substrates. As such, not only were skeletal metazoans more important contributors to reef building in the Ediacaran, but there were also more variable reef types with more complex ecologies, than previously thought. Such an abundance of inferred suspension feeders with biomineralised skeletons indicates the efficient exploitation of new resources, more active carbon removal with a strengthened energy flow between planktic and benthic realms, and the rise of biological control over benthic carbonate production. These mark the prelude to the Cambrian Explosion and the modernisation of the global carbon cycle.     

Dissolution of Dead Corals by Euendolithic Microorganisms Across the Northern Great Barrier Reef (Australia)

Spatial and temporal variabilities in species composition, abundance, distribution, and bioeroding activity of euendolithic microorganisms were investigated in experimental blocks of the massive coral Porites along an inshore–offshore transect across the northern Great Barrier Reef (Australia) over a 3-year period. Inshore reefs showed turbid and eutrophic waters, whereas the offshore reefs were characterized by oligotrophic waters. The euendolithic microorganisms and their ecological characteristics were studied using techniques of microscopy, petrographic sections, and image analysis. Results showed that euendolithic communities found in blocks of coral were mature. These communities were dominated by the chlorophyte Ostreobium quekettii, the cyanobacterium Plectonema terebrans, and fungi. O. quekettii was found to be the principal agent of microbioerosion, responsible for 70–90% of carbonate removal. In the offshore reefs, this oligophotic chlorophyte showed extensive systems of filaments that penetrated deep inside coral skeletons (up to 4.1 mm) eroding as much as 1 kg CaCO₃ eroded m⁻² year⁻¹. The percentage of colonization by euendolithic filaments at the surface of blocks did not vary significantly among sites, while their depths of penetration, especially that of O. quekettii (0.6–4.1 mm), increased significantly and gradually with the distance from the shore. Rates of microbioerosion (0.1–1.4 kg m⁻² after 1 year and 0.2–1.3 kg m⁻² after 3 years of exposure) showed a pattern similar to the one found for the depth of penetration of O. quekettii filaments. Accordingly, oligotrophic reefs had the highest rates of microbioerosion of up to 1.3 kg m⁻² year⁻¹, whereas the development of euendolithic communities in inshore reefs appeared to be limited by turbidity, high sedimentation rates, and low grazing pressure (rates <0.5 kg m⁻² after 3 years). Those results suggest that boring microorganisms, including O. quekettii, have a significant impact on the overall calcium carbonate budget of coral reef ecosystems, which varies according to environmental conditions.     

Effect of exogenous extracellular polysaccharides on the desiccation and freezing tolerance of rock-inhabiting phototrophic microorganisms

Two major stresses that threaten rock-inhabiting microbial communities are desiccation and freezing; both result in a loss of liquid water in the cells. The mechanisms necessary to tolerate these extremes may be similar, but are not well understood. In both cases extracellular polysaccharides (EPS) seem to play an important role. This study examines whether the EPS released by a rock-inhabiting phototroph can have a protective effect on other members of similar and neighboring microbial communities. This interaction was modeled by adding EPS isolated from the cryptoendolithic cyanobacterium Nostoc sp. to cells of the cryptoendolithic green alga Chlorella sp. and to cells of the epilithic cyanobacterium Chroococcidiopsis sp. The cells were then subjected to desiccation and freezing and the survival rates were determined by vital staining, using membrane integrity as a measure of viability. The results clearly demonstrate the importance of exogenous EPS in the desiccation tolerance of both species, while mixed results were found for the freezing trials.     

The ‘biomineralization toolkit’ and the origin of animal skeletons

Biomineralized skeletons are widespread in animals, and their origins can be traced to the latest Ediacaran or early Cambrian fossil record, in virtually all animal groups. The origin of animal skeletons is inextricably linked with the diversification of animal body plans and the dramatic changes in ecology and geosphere–biosphere interactions across the Ediacaran–Cambrian transition. This apparent independent acquisition of skeletons across diverse animal clades has been proposed to have been driven by co‐option of a conserved ancestral genetic toolkit in different lineages at the same time. This ‘biomineralization toolkit’ hypothesis makes predictions of the early evolution of the skeleton, predictions tested herein through a critical review of the evidence from both the fossil record and development of skeletons in extant organisms. Furthermore, the distribution of skeletons is here plotted against a time‐calibrated animal phylogeny, and the nature of the deep ancestors of biomineralizing animals interpolated using ancestral state reconstruction. All these lines of evidence point towards multiple instances of the evolution of biomineralization through the co‐option of an inherited organic skeleton and genetic toolkit followed by the stepwise acquisition of more complex skeletal tissues under tighter biological control. This not only supports the ‘biomineralization toolkit’ hypothesis but also provides a model for describing the evolution of complex biological systems across the Ediacaran–Cambrian transition.     

Fossil preservation in the Neoproterozoic Doushantuo phosphorite Lagerstätte, South China

Phosphorites of the late Neoproterozoic Doushantuo Formation exposed in the vicinity of Weng'an, Guizhou Province, and Chadian, Shaanxi Province, South China, contain exceptionally well-preserved algal thalli, acritarchs, and globular microfossils interpreted as animal embryos. Combined optical microscopic and SEM observations provide insights into the taphonomy of phosphatized fossils. Algal cells and tissues are variably resistant to decay, and within preserved populations permineralization began at varying stages of degradation. In consequence, there is a spectrum of quality in cellular preservation. Algal cell walls, acritarch vesicles, and embryo envelopes are commonly encrusted by an isopachous rim of apatite, with cell interiors filled by collophane and later diagenetic dolomite. In contrast, blastomere surfaces of animal embryos are encrusted primarily by minute phosphatic spherules and filaments, possibly reflecting an immediately postmortem infestation of bacteria that provided nucleation sites for phosphate crystal growth. Thus, the same processes that gave rise to Phanerozoic phosphatized Lagerstätten - phosphatic encrustation, and impregnation, probably mediated by microbial activity - effected soft-tissue preservation in the Doushantuo Lagerstätte. It remains unclear how phosphatic ions and organic macro-molecules interact at the molecular level and to what extent specific microbial metabolisms or microenvironmental conditions control the phosphatization of soft tissues. New observations of phosphatized Doushantuo fossils include: a second locality (Chadian) for Wengania globosa , interpreted as an algal thallus and previously known only from Weng'an; microtunnels in Weng'an phosphorites interpreted as pyrite trails; and new taxa described from Weng'an: Meghystrichosphaeridium reticulatum (acritarch), Sarcinophycus radiatus (algal thallus), and one unnamed problematic form.     

陕西西乡寒武纪幸运期宽川铺生物群网格状微体化石埋藏学与生物属性研究*

磷酸盐化保存是软躯体化石特异埋藏的一个重要途径, 而微生物在软躯体磷酸盐化过程中可能发挥了重要作用。前人通过埋藏学实验发现, 微生物会在动物胚胎等软躯体组织内部快速滋生, 充填生物体内部空间, 以微生物假形的方式复制了生物体的原始形态。但化石的磷酸盐化过程是否与埋藏学实验模拟的过程一致, 目前仍有争议。本次研究在寒武纪早期宽川铺生物群中发现了一类网格状微体化石。此类化石的保存状态可以根据其中丝状微生物滋生的程度分为三种类型, 它们展示了生物从死亡到微生物侵入、滋生, 最后被磷酸盐化的全过程。这些标本显示, 微生物假形在生物软组织磷酸盐化过程中扮演了重要角色, 但并不是化石磷酸盐化的必由之路, 尤其是当生物体具有矿化硬骨骼或者几丁质软骨骼等抗腐性较强的结构时。此类标本多以不完整保存的残片为主, 正反两面结构一致, 具有典型列状排列的近圆形与哑铃形网孔。由于化石结构简单, 生物学性状较少, 因此它们的亲缘关系尚不明确, 是一类需要继续研究的疑难化石。     

陕南晚震旦世骨骼化石中的微生物磷酸盐化作用

陕南晚震旦世高家山生物群中骨骼化石的电镜扫描显示,Sinotubulites levis壳壁中具有丰富的磷酸盐化球状和丝状体,可分为3种不同类型的微生物组构：1)由相互缠绕的丝状体所组成的藻席状组构;2)由外表面成莓球状,内部中空的球状体紧密堆叠所形成的组构类型;3)由表面极为光滑的球状体堆叠所形成的组构。它们可能分别代表不同类型的矿化蓝藻,依此推断微生物在S.levis壳壁磷酸盐化过程中起着重要作用。在共生的Sinotubulites baimatuoensis和Cloudina壳壁上,极少或完全不存在这些磷酸盐化组构表明微生物的磷酸盐化作用具有专属性,可能由壳壁微结构所决定,具多层状壁的S.levis可能与具许多有机质纹层的多毛类壳壁关系密切。     

Fossil preservation in the Neoproterozoic Doushantuo phosphorite Lagerstatte, South China.

Phosphorites of the late Neoproterozoic Doushantuo Formation exposed in the vicinity of Weng'an, Guizhou Province, and Chadian, Shaanxi Province, South China, contain exceptionally well-preserved algal thalli, acritarchs, and globular microfossils interpreted as animal embryos. Combined optical microscopic and SEM observations provide insights into the taphonomy of phosphatized fossils. Algal cells and tissues are variably resistant to decay, and within preserved populations permineralization began at varying stages of degradation. In consequence, there is a spectrum of quality in cellular preservation. Algal cell walls, acritarch vesicles, and embryo envelopes are commonly encrusted by an isopachous rim of apatite, with cell interiors filled by collophane and later diagenetic dolomite. In contrast, blastomere surfaces of animal embryos are encrusted primarily by minute phosphatic spherules and filaments, possibly reflecting an immediately postmortem infestation of bacteria that provided nucleation sites for phosphate crystal growth. Thus, the same processes that gave rise to Phanerozoic phosphatized Lagerstatten--phosphatic encrustation, and impregnation, probably mediated by microbial activity--effected soft-tissue preservation in the Doushantuo Lagerstatte. It remains unclear how phosphatic ions and organic macromolecules interact at the molecular level and to what extent specific microbial metabolisms or microenvironmental conditions control the phosphatization of soft tissues. New observations of phosphatized Doushantuo fossils include: a second locality (Chadian) for Wengania globosa, interpreted as an algal thallus and previously known only from Weng'an; microtunnels in Weng'an phosphorites interpreted as pyrite trails; and new taxa described from Weng'an: Meghystrichosphaeridium reticulatum (acritarch), Sarcinophycus radiatus (algal thallus), and one unnamed problematic form.     

Cambrian Series 3 lithistid sponge–microbial reefs in Shandong Province,North China: reef development after the disappearance of archaeocyaths

The Cambrian Series 3 Zhangxia Formation in Shandong Province, North China, includes small‐scale lithistid sponge–microbial reefs. The lithistid sponges grew on oolitic and bioclastic sediments, which were stabilized by microbial activities. The relative abundances of microbial components (e.g. calcimicrobe Epiphyton and stromatolites) vary among the reefs. However, the microbial components commonly encrusted or bound the lithistid sponges, formed remarkable encrustations on the surfaces of the sponges. Epiphyton especially grew upward and downward. The lithistid sponges thus provided substrates for the attachment and development of microbes, and the microbes played essential roles as consolidators, by encrusting reef‐building sponges. Additionally, the lithistid sponges were prone to degradation via microbial activities and diagenetic processes, and were thus preserved as micritic bodies, showing faint spicular networks or abundant spicules. Such low preservation potential within the reef environment obscured the presence of the sponges and their widespread contribution as reef‐building organisms during the Cambrian. During the prolonged interval after the demise of archaeocyaths, purely microbial reefs, such as stromatolites and thrombolites have been considered to be the principal reef builders, in association with rare lithistid sponge–microbial associations. However, recent findings, including those from Shandong Province and Korea, suggest that the lithistid sponge‐bearing reefs were more extensive during the Epoch 3 to the Furongian than previously thought. These lithistid sponge–microbial reefs were precursors of the sponge–microbial reefs that dominated worldwide in the Early Ordovician.     

Microboring organisms in living stylasterid corals (Cnidaria,Hydrozoa)

Microboring or euendolithic microorganisms, which colonize and penetrate various carbonate substrates, are abundant in coral reef ecosystems and play a major role in reef carbonate dissolution. A few studies reported the presence of euendoliths in stylasterid coral skeletons but the biological identity, distribution and abundance of these microorganisms remain largely unknown. Observations of over 100 stylasterid colonies, collected in the Indo-Pacific area, revealed for the first time that the association between these corals and euendolith organisms appears to be quite common in shallow tropical waters. The most abundant euendolith was identified as a cryptic stage in the development of the rhodophyte Porphyra (Conchocelis stage). The euendoliths were observed in the skeletons of seven species of three genera (four Stylaster, two Distichopora and one Lepidotheca). The presence of euendoliths inside skeletons conferred a particular colour to the studied stylasterid corals. Distribution and abundance of microborings varied significantly among stylasterid species and among branches of a single colony and so did the colour of their skeletons. Colonization of skeletons and the associated colour distribution were almost uniform in some stylasterids, forming an upward gradually diminishing or sharply limited gradient. This study shows that patterns of euendolith colonization and growth in stylasterid skeletons may depend on the stage of the euendolith development as well as on their environmental requirements such as light exposure.     

Osteological development of Cope's Gray Treefrog,Hyla chrysoscelis

Hyla chrysoscelis, Cope's Gray Treefrog, is a generalized treefrog found throughout much of east‐central North America. Although it is a model for many behavioural and ecological studies, little is known of its skeletal morphology or development. Herein, we describe the postembryonic skeletal development and adult osteology of H. chrysoscelis. The adult skull is well ossified with slight dermal ornamentation, the postcranial and tadpole skeletons are fairly non‐distinct with no obvious novel morphologies, and the Gosner stage by which bony elements first appear varies. We compare the rank order sequence of ossification to that of its sibling species Hyla versicolor and use examples from this study to demonstrate current complications with conducting ossification sequence meta‐analyses.     

Enrolment and trunk segmentation of a Cambrian eodiscoid trilobite

Completely enrolled, phosphatized, 3‐D specimens of the eodiscinid trilobite Tsunyidiscus yanjiazhaiensis from Cambrian Stage 3 of South China exhibit much morphological detail and show variation in delicate coaptative structures associated with their trunk segmentation. The relationship between enrolment mechanism and trunk segmentation during ontogeny confirms a unique developmental pattern among the early Cambrian eodiscoid trilobites, revealing how these animals controlled the rate of segment increase and release during post‐embryonic development in their life cycles.     

Fluidized muds: a novel setting for the generation of biosphere diversity through geologic time*

Reworked and fluidized fine‐grained deposits in energetic settings are a major modern‐day feature of river deltas and estuaries. Similar environments were probably settings for microbial evolution on the early Earth. These sedimentary systems act as efficient biogeochemical reactors with high bacterial phylogenetic diversity and functional redundancy. They are temporally rather than spatially structured, with repeated cycling of redox conditions and successive stages of microbial metabolic processes. Intense reworking of the fluidized bed entrains bacteria from varied habitats providing new, diverse genetic materials to contribute to horizontal gene transfer events and the creation of new bacterial ecotypes. These vast mud environments may act as exporters and promoters of biosphere diversity and novel adaptations, potentially on a globally important scale.     

Epilithic cyanobacterial communities of a marine tropical beach rock (Heron Island, Great Barrier Reef): diversity and diazotrophy

The diversity and nitrogenase activity of epilithic marine microbes in a Holocene beach rock (Heron Island, Great Barrier Reef, Australia) with a proposed biological calcification "microbialite" origin were examined. Partial 16S rRNA gene sequences from the dominant mat (a coherent and layered pink-pigmented community spread over the beach rock) and biofilms (nonstratified, differently pigmented microbial communities of small shallow depressions) were retrieved using denaturing gradient gel electrophoresis (DGGE), and a clone library was retrieved from the dominant mat. The 16S rRNA gene sequences and morphological analyses revealed heterogeneity in the cyanobacterial distribution patterns. The nonheterocystous filamentous genus Blennothrix sp., phylogenetically related to Lyngbya, dominated the mat together with unidentified nonheterocystous filaments of members of the Pseudanabaenaceae and the unicellular genus Chroococcidiopsis. The dominance and three-dimensional intertwined distribution of these organisms were confirmed by nonintrusive scanning microscopy. In contrast, the less pronounced biofilms were dominated by the heterocystous cyanobacterial genus Calothrix, two unicellular Entophysalis morphotypes, Lyngbya spp., and members of the Pseudanabaenaceae family. Cytophaga-Flavobacterium-Bacteroides and Alphaproteobacteria phylotypes were also retrieved from the beach rock. The microbial diversity of the dominant mat was accompanied by high nocturnal nitrogenase activities (as determined by in situ acetylene reduction assays). A new DGGE nifH gene optimization approach for cyanobacterial nitrogen fixers showed that the sequences retrieved from the dominant mat were related to nonheterocystous uncultured cyanobacterial phylotypes, only distantly related to sequences of nitrogen-fixing cultured cyanobacteria. These data stress the occurrence and importance of nonheterocystous epilithic cyanobacteria, and it is hypothesized that such epilithic cyanobacteria are the principal nitrogen fixers of the Heron Island beach rock.     

Moulting in the lobopodian Onychodictyon from the lower Cambrian of Greenland

A number of lobopodian taxa from the Cambrian display pairs of sclerotized plates symmetrically positioned along the dorsum of the animal, predominantly above the walking appendages. Most genera were described from complete body fossils exquisitely preserved in the famous Cambrian Lagerstätten, but lobopodian phosphatized plates are found worldwide as typical components of Cambrian small shelly fossil assemblages (SSF). Details regarding intraspecific and ontogenetic variation in lobopod plates are elusive, and the lack of details of ornamentation in Lagerstätte specimens does not minimize the problem. We document here an assemblage of well‐preserved isolated plates of Onychodictyon sp. from the Lower Cambrian (Cambrian Series 2, Stage 4) of North Greenland. Two specimens exhibit perfectly conjoined plates from successive moults. Details of ornamentation and the outline and profile of the fixed plates are identical, but width and length of the underlying plate are 24% larger. These specimens boost the body of evidence that lobopodians moulted but also show that plate outline and ornamentation did not vary during ontogeny.