Secondary sex characters appear in many heterotardigrades and a limited number of species of eutardigrades. In the eutardigrades Milnesium tardigradum and Pseudobiotus megalonyx, males are considered to always have modified claws on the first pair of legs; the basal branch of both claws in Milnesium and of the inner claw in Pseudobiotus is shaped like a robust hook. We examined one gonochoristic population of both Milnesium tardigradum and of Pseudobiotus megalonyx to evaluate the association of the modified claws with the presence of male germ cells in the testis. Three additional populations of Milnesium tardigradum were examined to provide data on the frequency of the modified claw among all individuals. In both species, molting animals were found with normal claws on the front legs of the old cuticle and modified claws on the front legs of the new cuticle. Examination of the gonad revealed that both species may have males with and without the modified claws. The appearance of the modified claw probably occurs at the last molt, and is used in mating when the male attaches to the female during copulation. Since this secondary sex character is not always present during the life of the tardigrade, but appears after a molt, a sex ratio determined only by the presence of the modified claw is not valid.

Tardigrades were collected from three seepage spring sites at 549m on Short Mountain, Cannon County, Tennessee. Three terrestrial samples and three aquatic samples were taken monthly from each site. Terrestrial samples included mosses and lichens from rocks and fallen trees; one sample of leaf litter was also collected from two of the sites. Aquatic samples included sediment, decaying leaf litter, and aquatic plants. Specimens of tardigrades were mounted on individual slides in Hoyer's medium. Species were identified and photographed using phase and differential contrast microscopy. Twenty-eight species representing 13 genera were recorded: Pseudechiniscus, Echiniscus, Doryphoribius, Ramazzottius, Hypsibius, Isohypsibius, Diphascon, Platicrista, Itaquascon, Murrayon, Macrobiotus, Minibiotus, and Milnesium. One of the Pseudechiniscus species and an Isohypsibius species were new to science and are described in this paper.

Background. The phylum Tardigrada consists of over 1,300 species that inhabit terrestrial, freshwater and marine environments throughout the world. In terrestrial habitats they live primarily in mosses, lichens, leaf litter and soil, whereas tardigrades in freshwater and marine environments are mainly found in sediments and on aquatic plants. More than 65 species have been previously reported in the state of Tennessee, USA. Methods. Tardigrades present in moss cushions (Grimmia sp.) collected from a xerothermic habitat on the East Tennessee State University campus, Johnson City, TN, USA, were extracted, mounted on slides, identified, and counted. Additional samples of fresh dried moss were used for integrative analyses, including morphological analysis with phase contrast (PCM) and scanning electron microscopy (SEM), as well as molecular analyses of COI, 18S rRNA, 28S rRNA, and ITS-2 of the Macrobiotus and Milnesium species. Results. Five species were found, including two species new to science: Viridiscus miraviridis sp. nov. and Macrobiotus basiatus sp. nov. Viridiscus miraviridis sp. nov. differs from other members of the genus mainly by having a different type of dorsal cuticle and some other, more subtle, morphometric characters. In addition to the two new species, Viridiscus perviridis and Viridiscus viridissimus were present, and males of Vir. viridissimus were found for the first time, the first record of males in the genus Viridiscus. Macrobiotus basiatus sp. nov. is most similar to Macrobiotus nelsonae, but it differs from Mac. nelsonae mainly by the stylet supports being situated in a more anterior position, shorter and narrower egg processes, and a smaller number of areoles around the egg processes. Moreover, the identification of Milnesium inceptum was confirmed as the first record for the USA by analysis of COI.

This monograph describes the global records of marine water bears (Phylum Tardigrada). We provide a comprehensive list of marine tardigrades recorded from around the world, providing an up-to-date taxonomy and a complete bibliography accompanied by geographic co-ordinates, habitat, substrate and biogeographic comments. A link is provided to an on-line interactive map where all occurrences for each species are shown. In total we list 197 taxa and their 2240 records from 39 oceans and seas and 18 Major Fishing Areas (FAO). It is hoped this work will serve as a reference point and background for further zoogeographic and taxonomic studies on marine tardigrades.

As part of the All Taxa Biodiversity Inventory (http://www.dlia.org), an extensive survey of tardigrades has been conducted in the Great Smoky Mountains National Park (GSMNP) in Tennessee and North Carolina, U.S.A., by Bartels and Nelson. Freshwater tardigrades include three species in the aquatic genus Thulinius (Eutardigrada, Isohypsibiidae). A new species, Thulinius romanoi, described from stream sediment, is distinguished from all other congeners by having a sculptured cuticle. In addition, the presence of Thulinius augusti (Murray, 1907) was verified by combined morphological and molecular analysis, and nine specimens of a third species, Thulinius cf. saltursus, were also found. Thulinius augusti is a new record for the United States. Thulinius saltursus (Schuster, Toftner & Grigarick, 1978) was previously recorded in California and Ohio, but our specimens vary slightly in morphology. The list of tardigrades from streams in the GSMNP was updated to a total of 44 species, 22 of which were predominantly or exclusively aquatic.

Based on the original species descriptions, a review of the genus Paramacrobiotus was conducted. We divided the genus into two subgenera, Microplacoidus subgen. nov. and Paramacrobiotus subgen. nov., based on the presence or absence of a microplacoid, characterized species within the genus based on seven different types of eggs. In a moss sample collected in Ecuador, Paramacrobiotus (Paramacrobiotus) spinosus sp. nov., was found. The new species differs from all species of the subgenus Parama crobiotus by the presence of richtersi type eggs and from other species by morphometric characters. Additionally, in the Ecuadorian material we found P. (Microplacoidus) magdalenae comb. nov., which is the first record of this species in Ecuador, we provide the full set of measurements for this species, not included in the original description. An additional new record is P. (M.) alekseevi comb. nov. found in Vietnam for the first time. After examining microscope slides from the Iharos' collection deposited in the Hungarian Natural History Museum, we prepared re-descriptions of P. (P.) csotiensis comb nov., P. (M.) submorulatus comb. nov. and P. (M.) wauensis comb. nov. Based on the morphological and morphometric characters of adults and eggs, we developed a diagnostic key to the genus Paramacrobiotus.