Key to sessile gnesiotrochan rotifers: Families, monospecific species in Flosculariidae, species of Atrochidae, Conochilidae, and Limnias.

Accurate identification of species is key to understanding their ecological roles and evolutionary history. It is also essential in cataloging biodiversity for comparisons among habitat types, responses to climate change, effective management practices, and more. The paucity of taxonomic expertise is increasing and with it the ability to competently identify species, this is particularly true for small taxa including rotifers. In an effort to improve this situation, we collated information on morphological characters from the literature on all valid species of sessile Gnesiotrocha (phylum Rotifera) currently assigned to two orders and four families. We review Order Collothecaceae, which comprises families Atrochidae (3 spp.) and Collothecidae (50 spp.) and Order Flosculariaceae, which includes families Conochilidae (7 spp.) and Flosculariidae (71 species). Based on that information, we provide dichotomous keys to the Families, monospecific species in Flosculariidae, and species of Atrochidae, Conochilidae, and Limnias. These keys will aid researchers to identify species in these families and lead to a better understanding of freshwater biodiversity and eco-evolutionary processes.

Ultrastructural characterization of the putative defensive glands (warts) in the sessile, colonial rotifer Sinantherina socialis (Gnesiotrocha; Flosculariidae).

Female Sinantherina socialis are freshwater, sessile, colonial rotifers that possess two pairs of distinctive glands (warts) located below the corona. Previous studies demonstrated that colonies are unpalatable to many invertebrate and vertebrate predators; those authors suggested that the warts were a possible source of a chemical deterrent to predation. Here we explore wart ultrastructure and cytochemisty to determine whether the warts function as exocrine glands and if their contents display any allomone-like chemistry, respectively. Externally, the warts appear as elevated bulges without pores. Internally, the warts are specialized regions of the integumental syncytium and therefore acellular. The lipid stain Nile Red labels all four warts. Two lipid membrane probes (sphingomyelin and phosphatidylinositol) also bind the warts and may be staining internal secretion vesicle membranes. In fact, wart ultrastructure is defined by hundreds of membrane-bound secretion vesicles packed tightly together. The vesicles are mostly electron-lucent and crowded into a well-defined cytoplasmic space. The cytoplasm also contains abundant ribosomes, rough endoplasmic reticulum, mitochondria, and Golgi, but nuclei are generally positioned peripheral to the packed vesicles. Absence of muscles around the warts or any signs of direct innervation suggests expulsion of gland contents is forced by general body contraction. A single specimen with 'empty' warts implies that secretions are released en masse from all glands simultaneously. The identity of the chemical secretion remains to be determined, but the lack of osmium and uranyl acetate staining suggests a low abundance or absence of phenols, unsaturated lipids, or NH2 and -COOH groups. This absence, combined with the positive Nile Red staining, is interpreted as evidence that vesicles contain saturated fatty acids such as lactones that are unpalatable to predators.

Genome size and lifestyle in gnesiotrochan rotifers.

Gnesiotrochan rotifers display a variety of life styles ranging from taxa with free-swimming larval and sessile adult stages to those with motile adult stages and colonial habits. Several explanations for the C- value enigma posits that genome size is correlated with lifestyle. To investigate this, 13 gnesiotrochan species representing nine genera were measured by flow cytometry. Genome sizes (1C) within Gnesiotrocha ranged from 0.05 pg (Hexarthra mira and Hexarthra fennica) to 0.25 pg (Sinantherina ariprepes). Genome sizes varied within genera and species; e.g., H. fennica (El Huérfano, Mexico) was estimated to be 15% larger than that of H. mira and H. fennica (Keystone Wetland, TX, USA). Gnesiotrochan genome sizes are similar to those reported within Ploima, which range from 0.06 pg (Brachionus rotundiformis, B. dimidiatus) to 0.46 pg (B. asplanchnoidis). Within Gnesiotrocha, genome size was found to be significantly smaller in sessile versus motile species as well as in solitary versus colonial species. To account for phylogenetic background, Linear Mixed Models with hierarchical taxonomic ranks showed that there is a taxonomic component underlying genome size. This study provides the first estimates of genome size within the superorder, providing a baseline for genomic and evolutionary studies within the group.

Fine structure of the subitaneous eggshell of the sessile rotifer Stephanoceros millsii (Monogononta) with observations on vesicle trafficking in the integument during ontogeny.

Rotifers that engage in cyclical parthenogenesis produce two types of eggs: subitaneous eggs that hatch as clonal females and meiotic eggs that hatch as haploid males, or if fertilized, as females after a period of diapause (resting eggs). The ultrastructure of resting eggshells is known for some motile species, but there are limited data on subitaneous eggshells, and no data on any eggshells of sessile rotifers. Here, we investigated the ultrastructure of the subitaneous eggshell of the sessile rotifer Stephanoceros millsii and its potential origins of secretion, the maternal vitellarium and embryonic integument. We also explored secretory activity in the larval and adult integuments to determine whether activity changes during ontogeny. The eggshell consists of a single layer with two sublayers: an external granular sublayer apparently derived from the maternal vitellarium, and an internal flocculent sublayer secreted by the embryonic integument that may form a hatching membrane or glycocalyx. Secretory activity remains high in both the larva and adult and appears to be the source of the thickening glycocalyx. Altogether, the subitaneous eggshell of S. millsii is the thinnest among monogonont rotifers. Thin eggshells may have evolved in response to the added protection provided by the mother's extracorporeal tube.

Limnological Characteristics and Rotifer Community Composition of Northern Mexico Chihuahuan Desert Springs.

Desert springs are critical for wildlife and human subsistence, but poorly studied. The springs in the northern Chihuahuan Desert region of Mexico are no exception. While groundwater overexploitation and other anthropogenic activities have endangered these waters, little is known about their water quality or biodiversity. We monitored five springs on a monthly basis for one year, and two others sporadically, to determine their physicochemical characteristics and to study rotifer community composition as an assessment of biodiversity. Seasonal variation in temperature, dissolved oxygen (DO), and conductivity was observed in the five springs. DO was occasionally below the criterion for aquatic life (<4mg/L) in some springs. Ca2+ (468-775.2 mg/L) was the dominant ion in all springs except Ojo de Santa María where Na+ (180-290 mg/L) dominated. Of 57 rotifer species identified, Lepadella patella (Müller, 1786), Lepadella triptera (Ehrenberg, 1830), and Philodina megalotrocha, Ehrenberg, 1832 were present in most springs. The rotifer community exhibited nestedness among the sites (supported by four null modes, p<0.05) and was correlated with fall season, DO, and nitrate concentration. This work provides information that will aid in management practices and provide important baseline data on water quality and regional diversity of a major component of the zooplankton.

Evidence supporting cryptic species within two sessile microinvertebrates, Limnias melicerta and L. ceratophylli (Rotifera, Gnesiotrocha).

Microorganisms, including rotifers, are thought to be capable of long distance dispersal. Therefore, they should show little population genetic structure due to high gene flow. Nevertheless, substantial genetic structure has been reported among populations of many taxa. In rotifers, genetic studies have focused on planktonic taxa leaving sessile groups largely unexplored. Here, we used COI gene and ITS region sequences to study genetic structure and delimit cryptic species in two sessile species (Limnias melicerta [32 populations]; L. ceratophylli [21 populations]). Among populations, ITS region sequences were less variable as compared to those of the COI gene (ITS; L. melicerta: 0-3.1% and L. ceratophylli: 0-4.4%; COI; L. melicerta: 0-22.7% and L. ceratophylli: 0-21.7%). Moreover, L. melicerta and L. ceratophylli were not resolved in phylogenetic analyses based on ITS sequences. Thus, we used COI sequences for species delimitation. Bayesian Species Delimitation detected nine putative cryptic species within L. melicerta and four putative cryptic species for L. ceratophylli. The genetic distance in the COI gene was 0-15.4% within cryptic species of L. melicerta and 0.5-0.6% within cryptic species of L. ceratophylli. Among cryptic species, COI genetic distance ranged 8.1-21.9% for L. melicerta and 15.1-21.2% for L. ceratophylli. The correlation between geographic and genetic distance was weak or lacking; thus geographic isolation cannot be considered a strong driver of genetic variation. In addition, geometric morphometric analyses of trophi did not show significant variation among cryptic species. In this study we used a conservative approach for species delimitation, yet we were able to show that species diversity in these sessile rotifers is underestimated.

Key to the currently recognized species of Limnias Schrank, 1803 (Rotifera, Monogononta, Gnesiotrocha, Flosculariidae).

Although the most widely used key to the Rotifera subsumes six species of the sessile genus Limnias within two species groups (L. ceratophylli and L. melicerta), the original descriptions of these forms are sufficiently different to recognize them as distinct entities. We used these descriptions and all available literature on these species to develop dichotomous and formula keys to the six species based on easily recognizable morphological characters. As part of our review we added relevant ecological information from published sources, as well as our own data. We also discuss the need for additional observations of morphological, behavioral, life history, and genetic features to better understand the diversity of this widespread genus.

Diaphanosoma Fischer, 1850 (Crustacea: Cladocera: Sididae) of Lake Turkana (East Africa), with the description of a new species of the genus.

Diaphanosoma turkanae sp. nov. is described from Lake Turkana (Kenya, East Africa). This species is the second Afrotropical endemic in the order Ctenopoda. It is characterized by a distinctive feature, the presence of an unusually small number of setae on the upper two-segmented antennal branch (exopodite); the proximal segment of the branch always bears three setae while the distal segment bears either six or seven setae. Morphologically the new species is most similar to D. orghidani Negrea, also occurring in Africa and presumably sharing an evolutionary affinity. D. turkanae sp. nov. co-occurs with two other species of the genus, D. lacustris Korinek and D. excisum Sars, which are briefly described here. Temperate-subtropical species of the genus Diaphanosoma penetrate far south along the Nile River system up to the equatorial and Ethiopian lakes and further southwards, often forming multi-species assemblages.

Soft Bodies, Hard Jaws: An Introduction to the Symposium, with Rotifers as Models of Jaw Diversity.

Jaws have evolved numerous times in the animal kingdom and they display a wide variety of structural, compositional, and functional characteristics that reflect their polyphyletic origins. Among soft-bodied invertebrates, jaws are known from annelids, chaetognaths, flatworms, gnathostomulids, micrognathozoans, mollusks, rotifers, and several ecdysozoans. Depending on the taxon, jaws may function in the capture of prey (e.g., chaetognaths and flatworms), processing of prey (e.g., gnathostomulids and onychophorans), or both (e.g., rotifers). Although structural diversity among invertebrates' jaws is becoming better characterized with the use of electron microscopy, many details remain poorly described, including neuromuscular control, elemental composition, and physical characteristics, such as hardness and resistance to wear. Unfortunately, absence of relevant data has impeded understanding of their functional diversity and evolutionary origins. With this symposium, we bring together researchers of disparately jawed taxa to draw structural and mechanistic comparisons among species to determine their commonalities. Additionally, we show that rotifers' jaws, which are perhaps the best-characterized jaws among invertebrates, are still enigmatic with regard to their origins and mechanics. Nevertheless, technologies such as energy dispersive X-ray spectroscopy (EDX) and 3D modeling are being used to characterize their chemical composition and to develop physical models that allow exploration of their mechanical properties, respectively. We predict that these methods can also be used to develop biomimetic and bioinspired constructs based on the full range of the complexity of jaws, and that such constructs also can be developed from other invertebrate taxa. These approaches may also shed light on common developmental and physiological processes that facilitate the evolution of invertebrates' jaws.

Lethal and sublethal effects of selected PPCPs on the freshwater rotifer, Plationus patulus.

Pharmaceuticals and personal care products (PPCPs) have been reported in surface waters around the world. The continuous input of these pollutants into freshwaters and their potential effects on aquatic life are of increasing concern. The rotifer Plationus patulus, a basal member of riverine food webs, was used to test acute and chronic toxicity of 4 PPCPs (acetamidophenol, caffeine, fluoxetine, triclosan). A population from a remote site in Mexico (reference population) and one from an urbanized stretch of the Rio Grande were exposed. Acute toxicity tests show that both populations were more sensitive to fluoxetine. Chronic exposure to acetamidophenol (10 mg/L, 15 mg/L, and 20 mg/L) inhibited reference population growth, whereas Rio Grande population growth was inhibited only at 15 mg/L and 20 mg/L. Population growth was inhibited at 200 mg/L and 300 mg/L of caffeine for both populations. Chronic exposure to fluoxetine (0.020 mg/L) significantly inhibited population growth for the Rio Grande population only. Triclosan (0.05 mg/L, 0.075 mg/L, 0.10 mg/L) had the most deleterious effects, significantly reducing both populations' growth rates. Sublethal effects of chronic exposure to PPCPs included decreased egg production and increased egg detachment. A mixed exposure (6 PPCPs, environmentally relevant concentrations) did not affect population growth in either population. However, the continuous introduction of a broad suite of PPCPs to aquatic ecosystems still may present a risk to aquatic communities. Environ Toxicol Chem 2015;34:913-922. © 2014 SETAC.

Sexual Reproductive Biology of a Colonial Rotifer Sinantherina socialis (Rotifera: Monogononta): Do mating strategies vary between colonial and solitary rotifer species?

In many aquatic invertebrates including monogonont rotifers, sex provides genetic variation and dormant stages that allows dispersal in time and space. While the reproductive biology of some solitary monogonont rotifer species is known, little is known concerning mating behaviors in colonial rotifers. Coloniality poses unique challenges to the typical mating behavior of solitary rotifers. For instance, most species engage in circling behavior, where the male swims in close proximity to the female. In colonial forms, access to a particular female may be hindered by nearby colony mates. Here we provide descriptions of (1) male morphology, (2) mating behavior, and (3) types of eggs of the widespread colonial rotifer Sinantherina socialis, and discuss modifications in mating strategies as a consequence of coloniality. Two important differences from mating patterns documented in solitary rotifers were found in S. socialis. First, duration of circling phase of mating is protracted for males encountering small colonies of females as compared to solitary females. Males encountering single females removed from their colonies behave similarly to those of solitary species. Second, duration of copulation in S. socialis is the shortest reported for any rotifer species. Endogamy might occur in this species as sons copulate with their sisters and mothers, at least under laboratory conditions. Examples of behaviour in linked video clips.

Genetic differentiation, behavioural reproductive isolation and mixis cues in three sibling species of monogonont rotifers.

Many aquatic species usually considered to be 'cosmopolitan' have been identified as cryptic species complexes, based on deep genetic differentiation. However, reproductive isolation among sibling cryptic species has rarely been studied, and interspecific hybridization is common in some taxa.We investigated isolation mechanisms and possible introgression among three cyclical parthenogenetic rotifer species in the Epiphanes senta complex that are found in very different freshwater habitats: temperate floodplains, subtropical desert rock pools and a tropical alpine lake. Whereas Epiphanes ukera is reproductively isolated from E. chihuahuaensis and E. hawaiiensis, the latter hybridize under laboratory conditions.While reproductive isolation is incomplete, RAPD profiles indicated unique genetic signatures and showed no evidence for introgression, indicating that these three species are diverging and have independent evolutionary trajectories.Testing cues for sexual reproduction in these cyclic parthenogens demonstrated that mixis in E. chihuahuaensis and E. ukera is influenced by population density, whereas E. hawaiiensis females rarely produce mictic offspring regardless of density. Different mixis cues are likely to separate sexual periods and effectively cause reproductive isolation between the species. Epiphanes ukera and E. chihuahuaensis males display mate guarding behaviour, and E. ukera males distinguish between conspecific and heterospecific females in mate choice experiments. Geographic isolation, along with different cues for mixis induction and mate recognition, act as reproductive barriers among these sibling species.

Rotifers from selected inland saline waters in the Chihuahuan Desert of México.

BACKGROUND: In spite of considerable efforts over past decades we still know relatively little regarding the biogeography of rotifers of inland waters in México. To help rectify this we undertook an extensive survey of the rotifer fauna of 48 water bodies in the Chihuahuan Desert of México. RESULTS: Of the sites surveyed, 21 had salinities >or= 2000 microS cm-1 and in these we found 57 species of monogonont rotifers and several bdelloids. Species richness in the saline sites varied widely, with a range in species richness of 1 to 27 and a mean (+/- 1SD) = 8.8 (+/- 6.2). Collectively all sites possess relatively high percent single- and doubletons, 33.3 and 21.7%, respectively. Simpson's Asymmetric Index indicated that similarity in rotifer species composition varied widely among a set of 10 sites. These were selected because they were sampled more frequently or represent unusual habitats. These SAI values ranged from 0.00 (complete dissimilarity) to 1.00 (complete similarity). The Jaccard Index varied between 0.00 and 0.35. This observation probably reflects similarities and differences in water chemistry among these sites. Inland saline systems differed in their chemical composition by region. Conductivity was related to hardness and alkalinity. In addition, hardness was positively associated with chloride and sulfate. RDA showed that several species were positively associated with chloride concentration. Other factors that were significantly associated with rotifer species included the presence of macrophytes, nitrate content, oxygen concentration, TDS, latitude and whether the habitat was a large lake or reservoir. CONCLUSION: This study illustrates the diversity of the rotiferan fauna of inland saline systems and the uniqueness among waterbodies. Conservation of these systems is needed to preserve these unique sources of biodiversity that include rotifers and the other endemic species found in association with them.

Interaction effects of multi-metal solutions (As, Cr, Cu, Ni, Pb and Zn) on life history traits in the rotifer Plationus patulus.

While many studies have determined toxicity of single heavy metals, few have addressed the combined effects of multiple elements at different concentrations. In order to estimate how metal mixtures affect population dynamics of freshwater rotifers, we exposed Plationus patulus to environmentally relevant concentrations and combinations of arsenic and five heavy metals (Cr, Cu, Ni, Pb and Zn) for 8 days. The life-history parameters, intrinsic rate of increase (r), relative cumulative reproduction (RCR), mortality ratio, mictic ratio, fertilization ratio, and number of deformed offspring were used as endpoints. Exposures were repeated 4 times and considered as blocks in the experimental design. Differences between treatments were evaluated through ANOVA and LSD mean analysis. The combined presence of As, Cu, Ni, Pb, and Zn in different concentrations produced a variety of changes in the population dynamics of P. patulus. However, only Cr produced a significant element*environment interaction. The presence of Cr leads to significant decreases in the intrinsic rate of increase and increased mortality ratio in multi-metal environments. The population growth (as r) decreased from 0.25 to -0.16 due to Cr (10 microg/L) interaction with the remaining elements, and values 0.16-0.12 were observed in treatments containing high levels (50 microg/L) of Cr. Mortality ratio increased from 0.066 to 0.822 due to Cr (10 and 50 microg/L) interactions with the remaining elements at high concentrations. Responses of this basal consumer to metal mixtures will provide insights into how aquatic food webs and their constituent organisms can be altered by anthropogenic inputs like heavy metal contamination.