Divergence and species discrimination in freshwater bryozoans (Bryozoa: Phylactolaemata)

We explored the suitability of nuclear and mitochondrial ribosomal markers [small subunit nuclear ribosomal RNA gene, large subunit nuclear ribosomal RNA gene, and a region spanning partial small mitochondrial ribosomal RNA subunit, four transfer RNA genes, and partial large mitochondrial ribosomal RNA subunit (referred to as rrnS‐rrnL)] for resolving patterns of diversification of 27 freshwater bryozoan species (class: Phylactolaemata) and evaluated the utility of statoblast ultrastructural features and molecular phylogenies for species discrimination in the Fredericellidae and Plumatellidae. Molecular data identified Plumatella fruticosa as distinct from the rest of the plumatellids, rendering the latter polyphyletic. rrnS‐rrnL was the most suitable marker for species discrimination and identified two undescribed species of Plumatella and at least two undescribed species of Fredericella. Lack of wide dispersal by fredericellid statoblasts may underlie the observed propensity for cryptic speciation and phylogeographical structure in Fredericella. Conversely, the strong dispersal potential of plumatellid statoblasts may mediate efficient gene flow between distant populations and explain the relatively low intraspecific divergence and lack of evidence for cryptic speciation. We show that species identification based on external features of statoblasts can be problematic in both genera, including for a putatively highly invasive, biofouling species, Plumatella vaihiriae, thereby highlighting the utility of rrnS‐rrnL sequences for species barcoding. © 2013 The Linnean Society of London     

Freshwater bryozoans (Bryozoa) of Norway III: distribution and ecology of Plumatella fruticosa

Bryozoans were investigated during field studies of 601 lakes and other surface water bodies throughout Norway from 1960 to 1978. The frequency of occurrence of Plumatella fruticosawas evaluated in relation to 12 environmental variables. Statistically significant deviations from the frequencies expected on the basis of random distribution were described using the categories preference, avoidance, and absence. According to our material P. fruticosais one of the most common bryozoan species in Norwegian fresh water, represented in samples from 251 localities. This species occurred frequently all over the country, north to 71° 06 N (the northernmost record globally). Maximum elevation above sea level was 1179 m (maximum for Northern Europe).In most studies of freshwater bryozoans from the holarctic region, Plumatella fruticosahas been reported as widely distributed, but locally rare. In Norway, however, P. fruticosa is frequently found all over the country, even far to the north. The species apparently finds an optimal climate in the cold temperate and cold regions of Norway. Plumatella fruticosapreferred lakes, avoided slow-flowing rivers and ponds, and was absent in smaller water bodies. The species preferred somewhat higher elevation (400–900 m above sea level), but avoided lakes with the lowest summer temperatures (below 11 °C). P. fruticosa also preferred oligotrophic conditions with poor aquatic vegetation, stony shores, lakes poor in calcium and magnesium (sometimes almost comparable with distilled water), where the water was clear and colourless, and slightly acidic to neutral. The species also preferred lakes surrounded by Sphagnum bogs and was absent when pH was below 5.2. P. fruticosa avoided eutrophic conditions with rich aquatic vegetation, alkaline water (pH above 7.0), lakes with a higher content of calcium and magnesium and those with strongly coloured water (above 100 mg Pt l^–1). In spite of its preference for ion-poor conditions, it was also found in a brackish water lake. For most environmental variables, the species had a wider tolerance range than reported from elsewhere.     

Freshwater bryozoans (Bryozoa) of Norway II: distribution and ecology of two species of Fredericella

Freshwater bryozoans were investigated during field studies of 601 lakes and other surface water bodies throughout Norway from 1960 to 1978. The frequency of occurrence of the two Fredericella species was evaluated in relation to 12 environmental variables. Statistically significant deviations from the frequencies expected on the basis of random distribution were described using the categories preference, avoidance and absence. The typical European Fredericella sultana (with glossy statoblasts) occurred at 72 localities, north to 70° 25 N (the northernmost record on a global scale). Its frequency dropped towards the north. Maximum elevation above sea level was 1097 m (maximum recorded for Northern Europe). Another species with pitted statoblasts, tentatively considered to be Fredericella indica, occurred at 50 localities, north to 71° 09 N (the northernmost record reported to date). The geographical distribution reflected a typical northern cold-water species. One third of the 50 records were from lakes situated north of the Arctic Circle, and rich colonies were found on the North Cape plateau, as far north as possible on the European mainland. The few records from southeastern Norway were from high mountain lakes, up to 1397 m above sea level (highest record for Europe). This species was not found in any of the numerous lowland lakes investigated in southeastern Norway where F. sultana was common. Both species tolerated a cold climate, especially Fredericella indica, and they occurred in a wide range of environments. F. sultana preferred lakes with rather low elevation above sea level, rich vegetation with plant species typical for eutrophic environments, gyttja sediments and stony "hard" shores, some wave action, high content of calcium, and slightly coloured water. This species avoided ponds and was absent from ditches and mires, avoided dystrophic lakes surrounded by Sphagnum bog, with dy sediments and strongly coloured water, avoided sites with small wave action, was indifferent to magnesium and low concentrations of calcium, and was absent from lakes with pH below 5.4. F. indica was only recorded from lakes. It preferred rather low temperatures, oligotrophic conditions with poor aquatic vegetation and stony shores, some wave action, pH above 7.0, and low water colour (below 11 mg Pt l^–1). This species avoided lakes with high temperatures and was absent above 20 °C, was absent where aquatic vegetation was rich or consisted of Sphagnum bog, avoided lakes with soft sediments of dygyttja and dy, places with small wave action, acid lakes, was absent at pH below 5.9, tended to avoid water with colour above 10 mg Pt l^–1 and was absent in lakes with colour above 40 mg Pt l^–1. F. sultana and F. indica occurred together in only six lakes, four in the north, and two in the southern mountain areas. The opposite sensitivity and reactions of the two species towards several environmental variables were supported by a logistic regression analysis. The main ecological difference between the two species is that F. indica appears to be more strongly dependent on climatic factors.     

Ultrastructure of the body cavities in Phylactolaemata (Bryozoa)

Only species belonging to the bryozoan subtaxon Phylactolaemata possess an epistome. To test whether there is a specific coelomic cavity inside the epistome, Fredericella sultana, Plumatella emarginata, and Lophopus crystallinus were studied on the ultrastructural level. In F. sultana and P. emarginata, the epistome contains a coelomic cavity. The cavity is confluent with the trunk coelom and lined by peritoneal and myoepithelial cells. The lophophore coelom extends into the tentacles and is connected to the trunk coelom by two weakly ciliated coelomic ducts on either side of the rectum. The lophophore coelom passes the epistome coelom on its anterior side. This region has traditionally been called the forked canal and hypothesized to represent the site of excretion. L. crystallinus lacks an epistome. It has a simple ciliated field where an epistome is situated in the other species. Underneath this field, the forked canal is situated. Compared with the other species, it is pronounced and exhibits a dense ciliation. Despite the occurrence of podocytes, which are prerequisites for a selected fluid transfer, there is no indication for an excretory function of the forked canal, especially as no excretory porus was found. J. Morphol. 2009. © 2008 Wiley‐Liss, Inc.     

Sequential development of Buddenbrockia plumatellae (Myxozoa: Malacosporea) within Plumatella repens (Bryozoa: Phylactolaemata)

Colonies of the freshwater bryozoan Plumatella repens collected from a river in the UK were found to be infected with the myxozoan parasite Buddenbrockia plumatellae following laboratory maintenance. Optimisation of the bryozoan diet allowed maintenance of infected colonies for 90 d, permitting observation by light and electron microscopy of the sequential parasitic developmental cycle. Parasite stages were associated with host peritoneum, identifying the primary developmental phase. The association of B. plumatellae cells with peritoneal basal lamina and morphological similarities between parasite and host suggested that the parasite remodelled host tissue. Progressive expansion and elongation of individual parasites led to the release of freely floating vermiform stages within the host coelomic cavities. Within these 'worms', intraluminal masses developed, resulting in the formation of spores. Upon maturation, the 'worms' ruptured, releasing many spores within the host that were subsequently discharged. Although parasitism led to increased bryozoan fragmentation and lowered statoblast production, some colonies did survive, resulting in repeated waves of infection. Long-term laboratory maintenance of infected bryozoan colonies could provide a means of maintaining B. plumatellae for study until the full life cycle is ascertained.     

Global diversity of bryozoans (Bryozoa or Ectoprocta) in freshwater

The present study considers 88 bryozoan species occurring in freshwater: 69 phylactolaemate and 19 gymnolaemate species. Roughly 49% of these species are confined to one zoogeographical region. The cosmopolitan status of species like Fredericella sultana, Plumatella repens or P. emarginata has to be reconsidered. Among the Phylactolaemata, which are phylogenetically older than the Gymnolaemata, the gelatinous species (Lophopodidae, Pectinatellidae, Cristatellidae) are more primitive than the branching tubular species (Plumatellidae, Fredericellidae). Guest editors: E. V. Balian, C. Lévêque, H. Segers & K. Martens Freshwater Animal Diversity Assessment     

Freshwater bryozoans (Bryozoa) of Norway IV: Distribution and ecology of four species of Plumatella with notes on Hyalinella punctata

Bryozoans were investigated during field studies of 601 lakes and other surface water bodies throughout Norway from 1960 to 1978. The frequency of occurrence of Plumatella repens was evaluated in relation to 12 environmental variables. Statistically significant deviations from the frequencies expected on the basis of random distribution were described using the categories preference, avoidance and absence. P. repens is one of the most common bryozoan species in Norway. It is represented at 227 localities in our material and by 29 additional records. The species occurred frequently all over the country, north to 71°09N (the northernmost record on the European mainland). Maximum elevation above sea level was 1052 m (maximum for Northern Europe). P. repens preferred lakes, avoided rivers, and was absent from the smallest water bodies. The species preferred (1) low elevation (2–200 m above sea level), (2) medium water temperature, (3) rich aquatic vegetation (eutrophic conditions), (4) small wave action, (5) high content of calcium and magnesium, (6) pH 6.4–9.6, and (7) rather high water colour. Otherwise P. repens avoided (1) habitats above 500 m but occurred up to 1052 m, (2) the lowest water temperature intervals, (3) sites with poor aquatic vegetation and stony shores, (4) medium wave action, (5) sites with a low calcium and magnesium content, (6) lakes of low water colour, and (7) lakes with pH below 6.6: it was absent from lakes with pH below 5.2. For many environmental variables, the species had a wider tolerance range than reported from elsewhere. P. fungosa, P. casmiana, and P. emarginata are all rare in Norway. The total numbers of known localities, with our records in (), are 18 (11), 7 (7), and 7 (5), respectively. These sites are all from lowland areas up to 133 m above sea level. Environmental variables in the habitats are described. P. fungosa was represented by a single floatoblast in each of the four northernmost lakes where it was recorded. One of these lakes was in North Norway some 1000 km away from the nearest known sites with living colonies. The presence of large colonies of P. fungosa in some parts of southern Norway and only `single floatoblast lakes' further north is discussed in relation to the equilibrium theory of island biogeography: birds facilitate immigration by carrying floatoblasts and environmental conditions prevent their germination and the development of living colonies. All records of P. casmiana and P. emarginata were from South Norway. Results for P. fruticosa have been published previously. Notes are given on an old and now missing sample from ca. 1900, identified by Lacourt (1968) as Hyalinella punctata – the only record of this species from Norway.     

Freshwater bryozoans (Bryozoa) of Norway V: review and comparative discussion of the distribution and ecology of the 10 species recorded

The paper is based on material presented in five contributions [Hydrobiologia 421:1–24, 2000; 459: 103–123, 2001; 479: 11–22, 2002; 501: 179–198, 2003; Archives de lInstitut grand-ducal de Luxembourg. Section des Sciences naturelles, physiques et mathématiques, Nouvelle Série 44: 127–143, 2002]. Nine species of Bryozoa were collected during field studies of 601 lakes and other surface water types in Norway. Five of the species occur throughout Norway (Plumatella fruticosa, P. repens, Paludicella articulata, Cristatella mucedo and Fredericella sultana) and one has a northerly range (Fredericella indica). With the exception of Plumatella repens, the northernmost records of these species globally are in Norway. All six species have been found at elevations of more than 1000m in South Norway, which is also their maximum elevation above sea level in Northern Europe. Three species (Plumatella fungosa, P. emarginata and P. casmiana) are rare and present only in lowland areas in South Norway. Lacourt [A Monograph of the Freshwater Bryozoa–Phylactolaemata. Zoologische Verhandelingen, Leiden 93: 1–159, 1968] considered an old and now mislaid sample from the same area to be Hyalinella punctata. Up to six species and an average of two were found in lakes. The mean numbers of species in ponds, and slow-flowing and rapid-flowing rivers were 0.6, 0.8 and 1.0, respectively. Co-occurrence of pairs of species in lakes was investigated. For 10pairs, the number of cases of co-occurrence was significantly higher than expected on the basis of random distribution. This was ascribed to some shared ecological requirements. Only one pair (Paludicella articulata–Plumatella fungosa) showed fewer cases of co-occurrence than expected. This was explained by differences in their geographical distribution and tolerance to ecological factors. Tolerance and other ecological responses were described for the six frequent species using two statistical methods. Method 1: frequency deviations, is based on the null hypothesis of random distribution. Ten environmental variables were studied (type of surface water, elevation above sea level, water temperature, aquatic vegetation, sediment, wave action, Ca, Mg, pH and water colour).The concepts absence, avoidance (frequency lower than expected), preference (frequency higher than expected) and presence (no significant deviation from expected frequency) were used. The degree of response of a particular species to a specific environmental variable was measured by the sum of the numerical values of the significant frequency deviations divided by the number of categories with significant deviations. The results can be used as adaptability indices or tolerance indices: the lower the index, the better a species is adapted for surviving with respect to the environmental variable in question. Diagrams showed (1) the relative strength of the response of the six frequent species, (2) the average index for each species for all variables and (3) the effect of each of the 10 environmental variables on the Bryozoa (the species treated as a unit: an ecological guild). Method 2: logistic regression, treated the presence or absence of each of the six frequent species as dependent variables using nine independent environmental variables. Comparison of the results of method 1 and 2, indicated that method 1, which gives details of how a species responds to different categories of a variable, was particularly useful.     

Biochemical genetics and taxonomy in Plumatella fungosa and P. repens (Bryozoa: Phylactolaemata)

SUMMARY. Some authors have recently concluded that, using morphological criteria, the freshwater bryozoans Plumatella repens and P. fugosa cannot be separated and should be regarded as conspecific. To test this conclusion, electrophoretic techniques have been used to examine genetic differences between the two nominate species at several enzyme loci. Significant variation at a malate dehydrogenase locus and three aminopeptidase loci establish beyond doubt that P. repens and P. fugosa are separate but related species. No significant variation was found between two populations of P. repens . Morphological characters for the distinction of the two species are discussed and evaluated.     

Biochemical genetics and taxonomy in Plumatella emarginata and P. repens (Bryozoa: Phylactolaemata)

SUMMARY. Although morphological distinctions are well documented, many authors have regarded the freshwater bryozoan Plumatella emarginata as a variety of P. repens. Differences between the two nominate species have been studied by the electrophoretic examination of genetic variation at enzyme loci. Genetic identity (Nei, 1972) was 0.286, a value well below that expected for conspecific allopatric populations. The results leave little doubt that specimens of the two species are from quite distinct and separate gene pools, with an unexpectedly high level of genetic differentiation. It is therefore concluded that P. emarginata is a valid species and is not merely a variety of P. repens. Morphological differences between the two species are also discussed.     

Ultrastructural study of metamorphosis in the freshwater bryozoan Plumatella fungosa (Bryozoa,Phylactolaemata)

Summary

At metamorphosis the attachment of the Plumatella larva to the substrate is effected by secretions from glandular cells in the apical plate, the leading pole during swimming. The larval mantle folds back and slides down towards the substrate. By ciliary activity an adhesive secretion is spread over the metamorphosing larva and the attachment area. Two polypides appear through the larval terminal opening. The mantle fold, together with gland cells, nerve cells, sensory cells, and muscle cells from the larva form a nutritive cell mass. Reduction of this nutritive cell mass is accomplished by autolysis and phagocytosis. An invaginated area of the nutritive cell mass is provided with a dense layer of microvilli, which seem to have an absorbtive function. The nutritive cell mass consisting of transitory larval tissues provides a significant source of nutrient for the developing polypide buds.     

Phylogenetic relationships of freshwater bryozoans (Ectoprocta, Phylactolaemata) inferred from mitochondrial ribosomal DNA sequences

Most species of freshwater bryozoans (Ectoprocta: Phylactolaemata) have few morphological distinctions, and within phylactolaemates, the morphology of the statoblast has been used to determine evolutionary relationships. Here, two controversial phylogenies have been proposed for phylactolaemates with regard to the relationship of Lophopodidae to other families. Two plumatellid genera, Gelatinella and Hyalinella , are candidates for the ancestral type of lophopodids. In addition, the coexistence of spines on the surfaces of the statoblast has led to the suggestion that lophopodids are closely related to the family Cristatellidae. In this study, we analysed mitochondrial DNA sequences of the 12S and 16S rDNA genes of 10 phylactolaemate species. Our results suggest that plumatellids may not be a direct ancestral group of lophopodids and that cristatellids are not a sister group of lophopodids. Fredericella , which was previously thought to be an ancestral group, was revealed to be derived. In addition, our results suggest that Stephanella is the most basal phylactolaemate. Mapping morphological characteristics onto the sequence-based phylogenetic tree revealed convergent evolution of statoblast characters.     

Transmission of Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea) to Fredericella sultana (Bryozoa: Phylactolaemata) by various fish species

Tetracapsuloides bryosalmonae is a myxozoan parasite of salmonids and freshwater bryozoans, which causes proliferative kidney disease (PKD) in the fish host. To test which fish species are able to transmit T. bryosalmonae to bryozoans, an infection experiment was conducted with 5 PKD-sensitive fish species from different genera. Rainbow trout Oncorhynchus mykiss, brown trout Salmo trutta, brook trout Salvelinus fontinalis, grayling Thymallus thymallus and northern pike Esox lucius were cohabitated with T. bryosalmonae-infected Fredericella sultana colonies and then subsequently cohabitated with statoblast-reared parasite free Bryozoa. Statoblasts from infected colonies were tested by PCR to detect cryptic stages of T. bryosalmonae, which may indicate vertical transmission of the parasite. In this study, brown trout and brook trout were able to infect Bryozoa, while there was no evidence that rainbow trout and grayling were able to do so. Few interstitial kidney stages of the parasite were detected by immunohistochemistry in brown trout and brook trout, while rainbow trout and grayling showed marked proliferation of renal interstitial tissue and macrophages with numerous parasite cells. Intraluminal stages in the kidney tubules were only detected in brown trout and rainbow trout. In contrast to previous observations, pike was not susceptible to PKD in these trials according to the results of T. bryosalmonae-specific PCR. No DNA of T. bryosalmonae was detected in any statoblast.     

Proliferative, presaccular stages of Tetracapsuloides bryosalmonae (Myxozoa: Malacosporea) within the invertebrate host Fredericella sultana (Bryozoa: Phylactolaemata)

Proliferative kidney disease (PKD), caused by the malacosporean parasite, Tetracapsuloides bryosalmonae, is a major disease of salmonid culture both in western Europe and North America. The fish are infected from spores that develop within freshwater bryozoans and are released into the water column. Although sporogenesis has been studied in the bryozoan host and occurs within sacs, the formation of these sacs from presaccular stages has only been hypothesized. Examination of infected bryozoans by using a range of techniques identified proliferating, presaccular amoeboid stages of T. bryosalmonae on the body wall of the bryozoan Fredericella sultana. These stages possessed unique electron-dense bodies and were observed as aggregating within the bryozoan metacoel, differentiating to form spore sacs. Spore sac growth was associated with the assimilation of the presaccular parasites rather than through cryptomitosis of sac mural cells. This sac formation through aggregation and assimilation suggests an intriguing mechanism by which T. bryosalmonae can cross-fertilize.     

New bryozoans (Bryozoa) from the Upper Devonian (Lower Famennian) of the Kuznetsk Basin

New bryozoans from the Upper Devonian (Famennian) of the borderlands of the Kuznetsk Coal Basin are described: Orthopora tomensis sp. nov. and Minussina incrustata sp. nov.     

Transoceanic ships as vectors for nonindigenous freshwater bryozoans

Aim The transport of organisms in ships’ ballast tanks is a dominant vector for aquatic invasions worldwide. Until recently, efforts to manage this vector have overlooked the potential transport of invertebrate resting stages in the residual waters and sediments within emptied ballast tanks, i.e. NOBOB (‘No Ballast On Board’) tanks. The resting stages (statoblasts) of freshwater bryozoans are often buoyant and locally abundant and thus can be taken up easily during ballasting operations. They are also resistant to extreme environmental conditions and can generate new colonies after being dormant for decades; as such, they would likely remain viable propagules after lengthy transport in ship ballast tanks. This study quantified the occurrence of freshwater bryozoan statoblasts in ballast tank sediments of transoceanic ships. Location North American Great Lakes. Methods We quantified the frequency of occurrence, abundance and diversity of bryozoans (as statoblasts) in residual sediment samples taken from 51 NOBOB tanks of 33 transoceanic ships visiting the Great Lakes from 2000 to 2002. Results Our study identified 11 species, comprising nearly 12% of the total number of freshwater bryozoans known worldwide. These include two exotic species unrecorded in the Great Lakes (Fredericella sultana and Lophopus crystallinus), an exotic species already established in the region (Lophopodella carteri) and three cosmopolitan species (Plumatella casmiana, P. fungosa and P. repens). Our estimates suggest that a ship with NOBOB tanks may carry up to 10⁶ statoblasts. Main conclusions The discovery of species unrecorded in the Great Lakes and the potentially large numbers of statoblasts being transported in ship ballast tanks indicate a significant risk of new species introductions. Furthermore, the presence of cosmopolitan species and an exotic species already established in the Great Lakes suggests the strong possibility of cryptic invasions via the introduction of exotic genotypes.     

Novel morphological and genetic tools to discriminate species among the family Plumatellidae (Phylactolaemata,Bryozoa)

Some species of the freshwater bryozoans (Bryozoa, Phylactolaemata) belonging to the genus Plumatella are remarkably difficult to identify because of the large similarity of superficial architecture of their statoblasts. The examination of statoblasts by scanning electron microscope (SEM) has in fact resolved only some taxonomic questions. In this article, the authors report on novel morphological and molecular traits to discriminate among ten species of Plumatellidae (P. viganoi, P. repens, P. geimermassardi, P. rugosa, P. reticulata, P. casmiana, P. fungosa, P. emarginata, P. vaihiriae, and Hyalinella punctata). The former traits are based on shape, number, and position of annular chamber pores, whereas the latter reside on amplification and sequence analysis of the Internal Transcribed Spacer (ITS) region of nuclear rDNA. The successful amplification of ITS region from statoblasts and zooidial tubules allowed us to sequence this region on all the species investigated. The ITS sequences showed the presence of sufficient and informative polymorphisms to discriminate among morphologically similar species. It is noteworthy that the resulting ITS phylogenetic tree largely corroborated the distinction of at least two groups of freshwater bryozoans inferred on the basis of the annular chamber pore morphology. This study provides innovative approaches to reliably characterize freshwater bryozoans species and gain more insight into their taxonomy, phylogenetic relationship, and biodiversity.