Myoanatomy of the marine tardigrade Halobiotus crispae (Eutardigrada: Hypsibiidae)

The muscular architecture of Halobiotus crispae (Eutardigrada: Hypsibiidae) was examined by means of fluorescent‐coupled phalloidin in combination with confocal laser scanning microscopy and computer‐aided three‐dimensional reconstruction, in addition to light microscopy (Nomarski), scanning electron microscopy, and transmission electron microscopy (TEM). The somatic musculature of H. crispae is composed of structurally independent muscle fibers, which can be divided into a dorsal, ventral, dorsoventral, and a lateral musculature. Moreover, a distinct leg musculature is found. The number and arrangement of muscles differ in each leg. Noticeably, the fourth leg contains much fewer muscles when compared with the other legs. Buccopharyngeal musculature (myoepithelial muscles), intestinal musculature, and cloacal musculature comprise the animal's visceral musculature. TEM of stylet and leg musculature revealed ultrastructural similarities between these two muscle groups. Furthermore, microtubules are found in the epidermal cells of both leg and stylet muscle attachments. This would indicate that the stylet and stylet glands are homologues to the claw and claw glands, respectively. When comparing with previously published data on both heterotardigrade and eutardigrade species, it becomes obvious that eutardigrades possess very similar numbers and arrangement of muscles, yet differ in a number of significant details of their myoanatomy. This study establishes a morphological framework for the use of muscular architecture in elucidating tardigrade phylogeny. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

Studies on the morphology and ultrastructure of the Malpighian tubules of Halobiotus crispae Kristensen, 1982 (Eutardigrada)

The excretory and osmoregulatory system of Halobiotus crispae consists of two lateral and one smaller dorsal Malpighian tubules, which empty into the digestive tract in the transition zone of the midgut and rectum. The tubules are identical at the ultrastructural level, and consist of an initial segment with three large cells, a thin transitional distal part lacking a nucleus, and a proximal part with 9–12 nuclei. The initial segment possesses deep basal infoldings and interdigitating, finger-shaped processes of the plasma membrane, large mitochondria and giant nuclei. The distal part is a short section which supports the initial segment. Cellular offshoots from the succeeding proximal part constitute the distal part. The distal and proximal parts contain intercellular canals with concretions of variable size. The exit of the proximal part into the digestive tract is characterized by the presence of microvilli. Correlated with the different stages in the cyclomorphosis of H. crispae , we observed size variation of the Malpighian tubules; thus, pseudosimplex stages have the largest tubules. We present suggestions concerning the physiology of the tubules and compare the Malpighian tubules of Tardigrada with the Malpighian papillae of Protura.     

Head sensory organs of Halobiotus stenostomus (Eutardigrada, Hypsibiidae)

The structure and arrangement of sensory organs in the tardigrade Halobiotus stenostomus (Richters 1908) have been studied using transmission and scanning electron microscopy techniques. The sensory organs found on the head of H. stenostomus are as follows: the circumoral sensory field, cephalic papillae, anterolateral and posterolateral sensory fields, and suboral sensory region. Four types of ciliated receptor structures are described in the sensory fields. The lateral sensory fields contain two types of receptor endings, dense and lucent, which differ in the presence or absence of a collar and in the structure of the outer dendrite segment. Two more types of receptor endings, ultrastructurally differing from the lateral sensory field receptors, are located in the suboral sensory region. Receptors with an asymmetric collar have been found, and a receptor ending without a collar is described for the first time in tardigrades. Unlike in other species studied, the sensory organs of H. stenostomus lack the lymph cavity surrounding the outer receptor segment. Similarity and differences in the ultrastructure of receptors between H. stenostomus and other species of Eutardigrada and Heterotardigrada are discussed.     

The first record of a rare marine tardigrade, Halobiotus crispae Kristensen, 1982 (Eutardigrada: Hypsibiidae), from the Svalbard Archipelago

A rare marine tardigrade Halobiotus crispae Kristensen, 1982 was found abundant in intertidal flats at Josephbukta, Bellsund fiord, West Spitsbergen. This is the first record of this species from the Svalbard Archipelago. So far, the species has been found only in seven other localities in the northern hemisphere (between 56° and 69°N). The present record greatly extends the known geographical range to the north and also indicates a much wider distribution of the species than formerly reported. Moreover, a potential indicative character of this species and of the study area in monitoring climatic changes is discussed.     

Morphometric analysis of Ramazzottius varieornatus (Hypsibiidae: Eutardigrada)

A sample from the holotype population of the limno-terrestrial eutardigrade Ramazzoltius varieornatus was examined in order to elucidate the variation occurring in the size of the sclerified structures of the pharyngeal apparatus. A frequency plot of body lengths was also produced to deduce the effectiveness of this technique in estimating the number of moults exhibited by this species. Problems associated with collecting and using morphometric data in this way are discussed. Additional ultrastructural information associated with moulting is also presented. New species within the genus may be difficult to recognize in the absence of adequate data related to intraspecific variation.     

Graphical Presentation of the African Tardigrade FaunaUsing GIS with the Description of Isohypsibius malawiensis sp. n. (Eutardigrada: Hypsibiidae) from Lake Malawi

The tardigrade fauna of the African continent is reviewed and presented graphically using Worldmap, a Geographical Information System (GIS) developed for exploring geographical diversity patterns in large biological datasets. References to the African tardigrade fauna have been gathered from published literature and supplemented with unpublished species information from the collection of Prof. Reinhardt M. Kristensen (RMK), Zoological Museum, University of Copenhagen. 156 species belonging to 36 genera of tardigrades are present. They consist of 105 eutardigrade species and 51 heterotardigrade species, of which 42 species are semiterrestrial and 9 species are marine. The presence of tardigrades are reported from 20 countries, but of these 9 are represented by a single reference. Marine tardigrades in particular have been neglected, with only a single report from the shores of the African continent. Data from the RMK collection of samples from Egypt is included in the analysis.

The scattered and sparse knowledge of the African tardigrade distribution makes general conclusions difficult, but emphasises the large regions which require further investigation. The current distribution patterns corresponds with easy accessible or ‘tourist’ locations. Regions where GIS could be used to illustrate ecological preferences are also pointed out by the analysis.

Isohypsibius malawiensis sp. n. is described from Lake Malawi. It has a smooth body surface and lacks eyes. The heteronych claws are without lunulae, but below the claws a double arched cuticular bar is present. The dorsal and ventral apophyses of the buccal tube have a large drop-shaped swelling. The first macroplacoid is the largest followed by the second, which is only slightly larger than the third; microplacoid is absent. Males are smaller than females. Females lay only one to two very large eggs in the exuvium. The Malpighian tubules are clover-shaped. The new species is interstitial in coarse sediment and no other tardigrade species were present.     

Gross brain morphology of Hypoptopomatinae and Neoplecostominae (Siluriformes: Loricariidae): Comparative anatomy and phylogenetic implications

Hypoptopomatinae and Neoplecostominae include about 250 valid species, a substantial portion of loricariid catfishes. Although the relationships among the members of these subfamilies have been inferred by many authors, the most recent hypotheses based on morphological and molecular data differ widely. Herein, we provide new data on the morphology of the central nervous system, and evaluate the usefulness of these characters in phylogenetic inference. To accomplish this, we characterized the gross brain morphology of those catfishes, and analyzed 54 neuroanatomical characters in a total of 40 terminal taxa representing Hypoptopomatinae and Neoplecostominae, and also members of Delturinae and Hypostominae as outgroups. Hypoptopomatinae and Neoplecostominae are recovered as separate subfamilies, and most of our results are compatible with morphology‐based analyses. We conclude that neuroanatomy provides an informative source of new characters with strong phylogenetic signal at all recovered taxonomic levels.     

Three new species of Diphascon of the pingue group (Eutardigrada,Hypsibiidae) from Antarctica

This paper contributes to the knowledge of the pingue group of species within the genus Diphascon. Three new species are described, Diphascon (D.) polare, D. (D.) dastychi and D. (D.) victoriae, from Victoria Land (Antarctica); they differ from other species of the pingue group in characters of the bucco-pharyngeal apparatus. D. polare and D. victoriae differ also in the claw shape. A key is provided to the Diphascon pingue group. Accepted: 30 November 1998     

Wing base structure supports Coleorrhyncha + Auchenorrhyncha (Insecta: Hemiptera)

The phylogenetic placement of the moss bugs (Insecta: Hemiptera: Coleorrhyncha) has been highly controversial. Many apparent morphological apomorphies support the close relationship between Coleorrhyncha and Heteroptera (=true bugs). However, a recent phylogenomic study strongly supported a sister group relationship between Coleorrhyncha and Auchenorrhyncha (planthoppers, leafhoppers, treehoppers, spittlebugs, and cicadas). To test these two alternative hypotheses, we examined the fore‐ and hindwing base structure of the only known extant macropterous species of Coleorrhyncha using binocular and confocal laser scanning microscopes and analyzed the data selected from the wing base phylogenetically. When full morphological data including the wing base characters were analyzed, the sister group relationship between Coleorrhyncha + Heteroptera was supported, agreeing with previous consensus based on morphology. In contrast, when only wing base characters were analyzed separately, the clade Coleorrhyncha + Auchenorrhyncha was recovered, in agreement with the result from the phylogenomic study. The membranous condition of the proximal median plate in the forewing was identified as a potential synapomorphy of the latter grouping, and the absence of the tegula was excluded as a potential synapomorphy of Coleorrhyncha and Heteroptera.     

The emergence of cetaceans: phylogenetic analysis of male social behaviour supports the Cetartiodactyla clade

The phylogeny of cetaceans is still unresolved. Two hypotheses prevail for the position of cetaceans among ungulates. The first hypothesis shows that Artiodactyla is monophyletic and is sister taxon to a clade composed of cetaceans and mesonychians. The second one shows that Artiodactyla is paraphyletic and contains Cetacea that is sister taxon of Hippopotamida. These hypotheses are based on fossil records and molecular studies. The behaviour of extant species can provide as much phylogenetic information as other classical parameters. I considered the behaviour observed during male agonistic interactions in placental mammals in order to determine which of these hypotheses was supported by the behaviour of extant species. Headbutting was only observed in ruminants, hippopotamids and cetaceans, supporting the paraphyletic nature of Artiodactyla. Primitive ruminants (tragulids) and two genera of ruminants (Moschus and Oreamnos) were not observed headbutting. These secondary losses were only present in 6.25% of the 48 surveyed ruminant genera. Head-to-head attacks emerged in pigs, which have developed dermal protusions. Yet, these confrontations are not based on mutual blow delivery. The behavioural evidence supports the inclusion of cetaceans in Artiodactyla.     

Brain anatomy of the marine tardigrade actinarctus doryphorus (arthrotardigrada)

Knowledge of tardigrade brain structure is important for resolving the phylogenetic relationships of Tardigrada. Here, we present new insight into the morphology of the brain in a marine arthrotardigrade, Actinarctus doryphorus, based on transmission electron microscopy, supported by scanning electron microscopy, conventional light microscopy as well as confocal laser scanning microscopy. Arthrotardigrades contain a large number of plesiomorphic characters and likely represent ancestral tardigrades. They often have segmented body outlines and each trunk segment, with its paired set of legs, may have up to five sensory appendages. Noticeably, the head carries numerous cephalic appendages that are structurally equivalent to the sensory appendages of the trunk segments. Our data reveal that the brain of A. doryphorus is partitioned into three paired lobes, and that these lobes exhibit a more pronounced separation as compared to that of eutardigrades. The first brain lobe in A. doryphorus is located anteriodorsally, with the second lobe just below it in an anterioventral position. Both of these two paired lobes are located anterior to the buccal tube. The third pair of brain lobes are situated posterioventrally to the first two lobes, and flank the buccal tube. In addition, A. doryphorus possesses a subpharyngeal ganglion, which is connected with the first of the four ventral trunk ganglia. The first and second brain lobes in A. doryphorus innervate the clavae and cirri of the head. The innervations of these structures indicate a homology between, respectively, the clavae and cirri of A. doryphorus and the temporalia and papilla cephalica of eutardigrades. The third brain lobes innervate the buccal lamella and the stylets as described for eutardigrades. Collectively, these findings suggest that the head region of extant tardigrades is the result of cephalization of multiple segments. Our results on the brain anatomy of Actinarctus doryphorus support the monophyly of Panarthropoda. J. Morphol. 275:173–190, 2014. © 2013 Wiley Periodicals, Inc.     

Autophagy as the cell survival in response to a microsporidian infection of the midgut epithelium of Isohypsibius granulifer granulifer (Eutardigrada: Hypsibiidae)

The midgut epithelial cells of many invertebrates may possess microorganisms which act as symbionts or pathogens (bacteria, microsporidia, viruses). During our previous studies on Isohypsibius granulifer granulifer Thulin, 1928 (Tardigrada, Eutardigrada), which examined alterations of the midgut epithelium during oogenesis, we found that some of the specimens were infected with microsporidia. All stages of pathogens occurred in the cytoplasm of the digestive cells in the midgut epithelium of I. g. granulifer that were infected with microsporidia: meronts, sporonts, sporoblasts, and spores. The cytoplasm of the digestive cells was rich in mitochondria, cisterns of rough endoplasmic reticulum (RER), and Golgi complexes. Autophagy in the digestive cells of the dorsal midgut was much more intensive in comparison with noninfected specimens. Membranes of phagophores surrounded the pathogens forming autophagosomes. These latter structures fused with lysosomes forming autolysosomes and residual bodies appeared. Neither glycogen granules nor droplets of varying electron density, which accumulated in digestive cells during vitellogenesis and choriogenesis, appeared in individuals with microsporidia. While the midgut epithelium in noninfected specimens takes part in vitellogenesis and choriogenesis, in infected specimens, midgut cells are involved in the process of autophagy as a survival strategy.     

Ultrastructural study of spermiogenesis and the testicular and spermathecal spermatozoon of the Gonochoristic Tardigrade Xerobiotus pseudohufelandi (Eutardigrada,Macrobiotidae)

This paper investigates by scanning and transmission electron microscopy spermiogenesis and spermatozoon morphology of the gonochoristic eutardigrade Xerobiotus pseudohufelandi (Macrobiotidae). During spermiogenesis clusters of spermatids are connected by cytoplasmic bridges that persist up to an advanced stage of maturation. Spermiogenesis is characterized by distinctive modifications of the nucleus and by the differentiation of an acrosome, tail and substantial midpiece. Testicular spermatozoa are folded with the hinge located between the head and midpiece, thus resembling a nut-cracker. The head includes a rod-shaped, bilayered acrosome and an elongated, helicoidal nucleus with condensed chromatin. The large kidney-shaped midpiece has hemispherical swellings/ovoid elements surrounding the centriole and an incomplete mitochondrial sleeve. The flagellum contains a ‘9+2’ axoneme and terminates in a tuft of microtubules. Spermathecal spermatozoa always have linear profiles. The acrosome and nucleus have the same morphological pattern as in testicular spermatozoa, whereas the midpiece is thin and lacks the hemispherical swellings, and the tail is reduced to a short stub. Functional considerations are presented, based upon this different morphology. Moreover, phyletic comparisons are made on the basis of sperm morphology, both for the family Macrobiotidae and the class Eutardigrada. J. Morphol. 234:11–24, 1997. © 1997 Wiley-Liss, Inc.     

On the fine structure of the creeping larva of Loxosomella murmanica: additional evidence for a clade of Kamptozoa (Entoprocta) and Mollusca

The creeping larva of the kamptozoan (entoproct) Loxosomella murmanica was investigated using transmission electron microscopy. The late larva exhibits a prominent apical organ connected to the ‘cerebral’ commissure of large cerebral ganglia, which supply the paired frontal organ. From the cerebral ganglia two paired nerve cords project backwards, closely resembling the tetraneuralian pattern of basal molluscs. In addition, a neural ring supplying the prototroch is present. The epidermis is composed of myoepithelial cells. Dorsally its cuticle is covered by granules of unknown composition. The prototroch consists of two ciliary rings; a downstream collecting system is not present. Although there is a one‐way gut with a lumen throughout, the larva obviously does not feed. A single pair of protonephridia is present. The foot sole shares distinct similarities with basic molluscs, particularly with those of the aplacophoran Solenogastres: The anterior part shows a huge, subepidermal pedal gland and several bundles of cirri consisting of compound cilia. The posterior part is ciliated with intraepithelial mucous cells interspersed. The dorsoventral muscle fibres show the mollusc‐like ventral intercrossing. The present results and previous findings, in particular the chitinous, non‐moulted cuticle, the sinus circulatory system, and a number of neural features shared by Kamptozoa and Mollusca, provide substantial evidence for a direct sister‐group relationship between these phyla. In addition, the basal position of the Solenogastres (Neomeniomorpha) within the Mollusca is corroborated.     

The mushroom bodies – prominent brain centres of arthropods and annelids with enigmatic evolutionary origin

Loesel, R. and Heuer, C.M. 2010. The mushroom bodies – prominent brain centres of arthropods and annelids with enigmatic evolutionary origin. —Acta Zoologica (Stockholm) 91 : 29–34 Mushroom bodies (MBs) are the most prominent and conspicuous neuropils in the brain of arthropods, onychophorans and vagile polychaete annelids but have not been described in any other animal group with complex brain architecture. Due to a number of unique neuroanatomical characters MBs can easily be identified and distinguished from other brain centres. However, their evolutionary origin and the question whether MBs are homologous structures is still under debate. This paper will briefly summarize the available morphological data and their implications with respect to the molecular evidence on early metazoan radiation. Unraveling the origin of MBs is an example of the challenges neurophylogenists will face in the future, especially so since it will signify a major step towards reconstructing early metazoan brain evolution.     

Greatly reduced phylogenetic structure in the cultivated potato clade (Solanum section Petota pro parte)

Premise of the Study

The species boundaries of wild and cultivated potatoes are controversial, with most of the taxonomic problems in the cultivated potato clade. We here provide the first in‐depth phylogenetic study of the cultivated potato clade to explore possible causes of these problems.

Methods

We examined 131 diploid accessions, using 12 nuclear orthologs, producing an aligned data set of 14,072 DNA characters, 2171 of which are parsimony‐informative. We analyzed the data to produce phylogenies and perform concordance analysis and goodness‐of‐fit tests.

Key Results

There is good phylogenetic structure in clades traditionally referred to as clade 1+2 (North and Central American diploid potatoes exclusive of Solanum verrucosum), clade 3, and a newly discovered basal clade, but drastically reduced phylogenetic structure in clade 4, the cultivated potato clade. The results highlight a clade of species in South America not shown before, ‘neocardenasii’, sister to clade 1+2, that possesses key morphological traits typical of diploids in Mexico and Central America. Goodness‐of‐fit tests suggest potential hybridization between some species of the cultivated potato clade. However, we do not have enough phylogenetic signal with the data at hand to explicitly estimate such hybridization events with species networks methods.

Conclusions

We document the close relationships of many of the species in the cultivated potato clade, provide insight into the cause of their taxonomic problems, and support the recent reduction of species in this clade. The discovery of the neocardenasii clade forces a reevaluation of a hypothesis that section Petota originated in Mexico and Central America.