Ovarian Structure in Milnesium tardigradum (Tardigrada, Milnesiidae) during Early Vitellogenesis

The ultrastructure of the ovary of Milnesium tardigradum during early vitellogenesis is described. Within the ovary, there were large multinuclear cells surrounded by many mononuclear oocytes. Observation of serial sections revealed four multinuclear cells that were connected to each other by cytoplasmic bridges. Each peripheral oocyte was connected to the multinuclear cell. An enormous ER-like structure was conspicuous in the centre of the multinuclear cell. The presence of large numbers of lipid droplets and yolk granules in both multinuclear cells and many mononuclear oocytes suggested a role as nurse cells. A small number of these oocytes grow to be eggs. The structural features of the multinuclear nurse cell were compared with other known examples.     

The cerebral ganglia of Milnesium tardigradum Doyère (Apochela, Tardigrada): Three dimensional reconstruction and notes on their ultrastructure

Differential interference contrast micrographs from stretched animals, serially sectioned semi-thin and ultrathin sections revealed that the cerebral ganglia (supraoesophageal mass) of the eulardigrade Milnesium tardigradum lie above the buccal tube and adjacent tissue like a saddle. It has an anterior indentation which is penetrated by two muscles that arise from the cuticle of the forehead. The cerebral ganglia consist of lateral outer lobes bearing an eye on each side, and two inner lobes which extend caudally. Between the inner lobes a cone-like projection tapers into a nerve bundle. Each outer lobe is joined with the first ventral ganglion. From the outer lobe near the eye the ganglion for a posterolateral sensory field extends to the epidermis. Anterior to the supraoesophageal mass are three dorsal ganglia for the upper three peribuccal papillae. Two additional ganglia attached to the cerebral mass supply the lateral cephalic papillae. The cerebral ganglia are covered by a thin neural lamella. The pericarya which surround the neuropil have large nuclei. Near the axons in the centre of the supraoesophageal mass the cytoplasm is crowded with vesicles of different size and appearance. Some of them resemble synaptic vesicles while others resemble dense core bodies. Structurally different types of synapses and axons can be distinguished within the neuropil.     

Comparative proteome analysis of Milnesium tardigradum in early embryonic state versus adults in active and anhydrobiotic state

Tardigrades have fascinated researchers for more than 300 years because of their extraordinary capability to undergo cryptobiosis and survive extreme environmental conditions. However, the survival mechanisms of tardigrades are still poorly understood mainly due to the absence of detailed knowledge about the proteome and genome of these organisms. Our study was intended to provide a basis for the functional characterization of expressed proteins in different states of tardigrades. High-throughput, high-accuracy proteomics in combination with a newly developed tardigrade specific protein database resulted in the identification of more than 3000 proteins in three different states: early embryonic state and adult animals in active and anhydrobiotic state. This comprehensive proteome resource includes protein families such as chaperones, antioxidants, ribosomal proteins, cytoskeletal proteins, transporters, protein channels, nutrient reservoirs, and developmental proteins. A comparative analysis of protein families in the different states was performed by calculating the exponentially modified protein abundance index which classifies proteins in major and minor components. This is the first step to analyzing the proteins involved in early embryonic development, and furthermore proteins which might play an important role in the transition into the anhydrobiotic state.     

A new species of Tardigrada (Eutardigrada: Milnesiidae): Milnesium krzysztofi from Costa Rica (Central America)

Abstract

A new eutardigrade species Milnesium krzysztofi sp. nov. is described from Costa Rica. M. krzysztofi sp. nov. differs from the most similar Milnesium katarzynae Kaczmarek et al., 2004 mainly by the presence of spurs on internal claws I–III and on external claw IV.     

Studies on the tardigrades. 3. Fine structure of the esophagus of Milnesium tardigradum Doyère

A fine structural analysis of the esophagus of the tardigrade Milnesium tardigradum Doyère is presented. This foregut structure exhibits pronounced secretory activity with discharge of accumulated material by fusion of secretory vesicles with luminal and lateral surfaces of the esophageal cells. The possible function of this secretory material as well as the organization of esophageal cells and structure of the cuticular lining are discussed.     

Studies on the tardigrades. IV. Fine structure of the hindgut of Milnesium tardigradum doyère

The hindgut of the semi-terrestrial tardigrade, Milnesium tardigradum was examined with light and electron microscopy. The hindgut consists of a cloaca and an anterior hindgut. It is delineated anteriorly by the pylorus into which four Malpighian tubules empty and posteriorly, by a broad cloacal slit. A single oviduct enters the hindgut at the junction between the cloaca and the anterior hindgut. Two pairs of muscles insert on the cloaca and anterior hindgut respectively. Electron microscopic observations demonstrate that the anterior hindgut is a specialized transporting epithelium. The luminal surface is covered by a thin layer of cuticle which penetrates into channel-like invaginations. Numerous mitochondria are concentrated apically. The basal and lateral surfaces are also folded. The cells are joined apically by deep tight junctions and a simple basal lamina lines the entire hindgut. The cloaca which receives the contents of the gut and Malpighian tubules as well as gametes of the reproductive tract is a transitional organ that exhibits several characteristics of the hypodermis and anterior hindgut. The cuticle of the cloaca changes sequentially from the complex structure of the integument to a simple layer of the anterior hindgut. The function of the hindgut is discussed with emphasis on the possible response of the anterior hindgut to a hypoosmotic habitat, evaporative water loss during the induction of anhydrobiosis and low oxygen tension.     

Desiccation tolerance of the tardigrade Milnesium tardigradum collected in Sapporo, Japan, and Bogor, Indonesia

A tardigrade Milnesium tardigradum showed anhydrobiotic capacity, in which the desiccation tolerance, given by the mean survival rate under desiccation at different relative humidity levels, was significantly higher in the Sapporo (Japan) population than that in the Bogor (Indonesia) population. Accordingly, the surviving tardigrades took a significantly longer time for revival in Bogor than those in Sapporo. The higher tolerance of the Sapporo population is thought to be related to the low relative humidity and low temperature such that the animals experience 41% RH in May and often -10 degrees C or lower in winter.     

Milnesium cfr. tardigradum (Milnesiidae, Apochela, Tardigrada): A Monitor of High Altitude Meiofauna on Sub-Antarctic Marion Island

The meiofaunal tardigrades Milnesium cfr. tardigradum, Echiniscus sp., and Macrobiotus sp. were extracted from high altitude moss and volcanic scoria samples on sub-Antarctic Marion Island. Gut contents of mounted Milnesium provided forensic evidence of a previously ignored diverse and abundant meiofauna including trophi of bdelloid rotifers, mouthparts and entire individuals of Diphascon sp. (Tardigrada).     

Molecular phylogeny of Arthrotardigrada (Tardigrada)

Tardigrades are microscopic ecdysozoans with a worldwide distribution covering marine, limnic and terrestrial habitats. They are regarded as a neglected phylum with regard to studies of their phylogeny. During the last decade molecular data have been included in the investigation of tardigrades. However, the marine arthrotardigrades are still poorly sampled due to their relative rarity, difficult identification and minute size even for tardigrades. In the present study, we have sampled various arthrotardigrades and sequenced the 18S and partial 28S ribosomal subunits. The phylogenetic analyses based on Bayesian inference and maximum parsimony inferred Heterotardigrada (Arthrotardigrada + Echiniscoidea) and Eutardigrada to be monophyletic. Arthrotardigrada was inferred to be paraphyletic as the monophyletic Echiniscoidea is included within the arthrotardigrades. The phylogenetic positions of Stygarctidae and Batillipedidae are poorly resolved with low branch support. The Halechiniscidae is inferred to be polyphyletic as the currently recognized Styraconyxinae is not part of the family. Archechiniscus is the sister-group to the Halechiniscidae and Orzeliscus is placed as one of the basal halechiniscids. The phylogeny of the included eutardigrade taxa resembles the current molecular phylogenies. The genetic diversity within Arthrotardigrada is much larger (18S 15.1–26.5%, 28S 7.2–20.7%) than within Eutardigrada (18S 1.0–12.6%, 28S 1.3–8.2%). This can be explained by higher substitution rates in the arthrotardigrades or by a much younger evolutionary age of the sampled eutardigrades.     

Global diversity of tardigrades (Tardigrada) in freshwater

Tardigrada is a phylum closely allied with the arthropods. They are usually less than 0.5 mm in length, have four pairs of lobe-like legs and are either carnivorous or feed on plant material. Most of the 900+ described tardigrade species are limnoterrestrial and live in the thin film of water on the surface of moss, lichens, algae, and other plants and depend on water to remain active and complete their life cycle. In this review of 910 tardigrade species, only 62 species representing13 genera are truly aquatic and not found in limnoterrestrial habitats although many other genera contain limnoterrestrial species occasionally found in freshwater. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Diet of Physalaemus cf. cicada (Leptodactylidae) and Bufo granulosus (Bufonidae) in a semideciduous forest.

We determined the diet of the two most abundant anuran species which occur in the litter of a semideciduous forest (Len?óis, Bahia, Brazil), Physalaemus cf. cicada and Bufo granulosus in the dry and rainy seasons. Pitfall traps were used to collect anuran and invertebrate fauna, which showed the availability of prey in the environment. Physalaemus cf. cicada was present in both seasons and Bufo granulosus only in the rainy season. Both species fed mainly on Isoptera and Formicidae. However, there is a difference between the rainy and dry seasons concerning the diet of P. cf. cicada. During the rainy season P. cf. cicada consumed less Isoptera and more Formicidae than in the dry season. In the volumetric sense, Orthoptera was the most important alimentary category for P. cf. cicada and B. granulosus. The Jacobs electivity index indicated that Physalaemus cf. cicada and Bufo granulosus were specialists in Isoptera.     

Maintenance and loss of heterozygosity in a thelytokous lineage of honey bees (Apis mellifera capensis)

An asexual lineage that reproduces by automictic thelytokous parthenogenesis has a problem: rapid loss of heterozygosity resulting in effective inbreeding. Thus, the circumstances under which rare asexual lineages thrive provide insights into the trade-offs that shape the evolution of alternative reproductive strategies across taxa. A socially parasitic lineage of the Cape honey bee, Apis mellifera capensis, provides an example of a thelytokous lineage that has endured for over two decades. It has been proposed that cytological adaptations slow the loss of heterozygosity in this lineage. However, we show that heterozygosity at the complementary sex determining (csd) locus is maintained via selection against homozygous diploid males that arise from recombination. Further, because zygosity is correlated across the genome, it appears that selection against diploid males reduces loss of homozygosity at other loci. Selection against homozygotes at csd results in substantial genetic load, so that if a thelytokous lineage is to endure, unusual ecological circumstances must exist in which asexuality permits such a high degree of fecundity that the genetic load can be tolerated. Without these ecological circumstances, sex will triumph over asexuality. In A. m. capensis, these conditions are provided by the parasitic interaction with its conspecific host, Apis mellifera scutellata.     

Natal Dispersal Patterns of a Subsocial Spider Anelosimus cf. jucundus (Theridiidae)

Species that alternate periods of solitary and social living may provide clues to the conditions that favor sociality. Social spiders probably originated from subsocial‐like ancestors, species in which siblings remain together for part of their life cycle but disperse prior to mating. Exploring the factors that lead to dispersal in subsocial species, but allow the development of large multigenerational colonies in social species, may provide insight into this transition. We studied the natal dispersal patterns of a subsocial spider, Anelosimus cf. jucundus, in Southeastern Arizona. In this population, spiders disperse from their natal nests in their penultimate and antepenultimate instars over a 3‐mo period. We tracked the natal dispersal of marked spiders at sites with clustered vs. isolated nests. We found that most spiders initially dispersed less than 5 m from their natal nests. Males and females, and spiders in patches with different densities of nests, dispersed similar distances. The fact that both sexes in a group dispersed, the lack of a sex difference in dispersal distance, and the relatively short distances dispersed are consistent with the hypothesis that natal dispersal results from resource competition within the natal nest, rather than inbreeding avoidance in competition for mates. Additionally, an increase in the average distance dispersed with time and with the number of spiders leaving a nest suggests that competition for nest sites in the vicinity of the natal nest may affect dispersal distances. The similar distances dispersed in patches with isolated vs. clustered nests, in contrast, suggest that competition among dispersers from different nests may not affect dispersal distances.     

Ultrastructure of spermiogenesis and spermatozoa of Gymnotus cf. anguillaris and Brachyhypopomus cf. pinnicaudatus (Teleostei: Gymnotiformes)

The ultrastructure of spermiogenic stages and spermatozoa of representatives of two gymnotiform families, Gymnotus cf. anguillaris (Gymnotidae) and Brachyhypopomus cf. pinnicaudatus (Hypopomidae) were studied. Spermiogenesis of both species is characterized by lateral development of the flagellum and formation of a nuclear fossa. Some differences were found between these species, such as whether (B. cf. pinnicaudatus) or not (G. cf. anguillaris) nuclear rotation occurs, permanence of the cytoplasmic channel, and type and localization of the nuclear fossa. In the G. cf. anguillaris spermatozoon the nucleus is spherical with highly condensed chromatin. The nuclear fossa is shallow and lateral and is associated with the centriolar complex through stabilizing fibrils. The midpiece is short, with many vesicles, a cytoplasmic channel, and elongate mitochondria. In the B. cf. pinnicaudatus spermatozoon the ovoid nucleus is elongated lateral and posterior to the centriolar complex, and has highly condensed chromatin. The eccentric nuclear fossa is of the moderate type, and contains the entire centriolar complex. The midpiece is long, with numerous vesicles, elongate mitochondria, and no cytoplasmic channel. In both species the flagella are laterally disposed in relation to the nucleus and comprise of the classical 9+2 axoneme. Most of the characteristics found in the spermatozoa of these two species of Gymnotiformes are shared with species of Characiformes, whereas only a few are also found in Siluriformes. This suggests that Gymnotiformes and Characiformes may be more closely related than previously proposed.     

Micro-distribution of Soil Inhabiting Tardigrades (Tardigrada) in a Sub-alpine Coniferous Forest of Japan

Micro-distribution of soil inhabiting tardigrades was studied in a sub-alpine coniferous forest (alt. 1,840 m a. s. l.) on the eastern slope of Mt. Yatsugatake, Central Japan. Eight cylindrical soil samples were taken from the forest floor. Each sample was divided into 10 slice samples of 1 cm thick. Tardigrades were extracted from samples by Baermann funnels, identified to specific level and counted. The average number of tardigrades in the study site is 74,058/m². High densities sometimes occurred at depths greater than 5 cm. Consequently for investigating tardigrade populations, 5cm depth core sampling is insufficient in this type of habitat. Composition of the main groups of species reveals that shallow layers (1, 2 or 3 cm depth) are frequently dominated by the Diphascon-group. Macrobiotus-group species generally occurred in higher abundance in the deeper layers. It is very interesting that there are some different patterns of soil micro-distribution in tardigrade species.     

A new type of male dimorphism with ergatoid and short-winged males in Cardiocondyla cf. kagutsuchi

Summary. A new type of ant male dimorphism, consisting of wingless (ergatoid) and short-winged (brachypterous) males, was found in a species of the “Cardiocondyla kagutsuchi”- complex from Malaysia. The ergatoid males show the typical morphological and behavioral characteristics of those of many other Cardiocondyla species. The brachypterous males are morphologically intermediate between ergatoid males and typical winged males of other taxa in this genus. On one hand, they share a number of morphological and behavioral features with ergatoid males that might be adaptations to the loss of flight and intranidal mating: aggressive behavior towards rival males, a prolonged spermatogenesis, which is unique in winged males, paler body coloration, smaller compound eyes, shorter antennal funiculi, more rounded heads – perhaps due to the increased development of mandibular muscles, and an angular pronotum, probably for neck protection. Their short wings appear to protect the petiolar joints during fighting. On the other hand, the brachypterous males have not become as specialized as the ergatoids and to some extent keep the nature of the winged males of other species, i.e., they escape from the nest with a higher probability and with less injuries and do not show a reduction of the ocelli. In the sexual production season, the ergatoid males emerged first in small numbers and then both male morphs emerged in large numbers. The sex ratio was extremely female-biased in the earlier stage of sexual production, probably due to local mate competition.Received 13 December 2004; revised 17 February 2005; accepted 22 February 2005.     

Induction of males in thelytokous populations of Encarsia meritoria and Encarsia protransvena: A systematic tool

Males from thelytokous females of Italian uniparentalpopulations of Encarsia meritoria and Encarsiaprotransvena were obtained by feeding wasps with tetracycline orrearing parasitoids at high temperatures respectively. Two thermictreatments were tested on E. protransvena: 31°C constanttemperature versus an alternation of 12 hours at 33°C and 23°Con a daily basis. Females of E. meritoria fed with tetracyclineproduced 80.87% of male progeny. Tetracycline did not induce maleoffspring in E. protransvena. For this species hightemperatures were required even though only a few males were obtained(0.31–5.43%). These results confirm the infection ofparthenogenesis-inducing micro-organisms in E. meritoria, whilethis is doubtful in E. protransvena. Males derived fromthelytokous populations of the two species were described. Males of theuniparental population of E. meritoria were similar to those ofbiparental ones, suggesting that the two populations are conspecific.Males of E. protransvena are slightly different from thosedescribed previously, supporting the hypothesis that E.protransvena is a complex of cryptic species. Our results suggestthe possibility of using ``induced males' by either antibiotics or hightemperatures as a taxonomic tool in the genus Encarsia toseparate similar (if not undistinguishable) uniparentalspecies.