Comparative studies on the statoblasts of higher phylactolaemate bryozoans

Statoblasts of five higher phylactolaemates were compared morphologically. As a result, they were divided into two groups: Group I comprising Lophopus crystallinus, Lophopodella carteri, and Pectinatella gelatinosa, and Group II comprising Pectinatella magnifica and Cristatella mucedo. These two groups are thought to represent independent evolutionary series. In Group I and in P. magnifica, the statoblasts are curved to varying degrees after the manner of a saddle. When the dorsal and ventral valves are flattened, therefore, the contour is different between the two. In Group I, the outermost layer of a mature statoblast is hard-gelatinous and basophilic; it remains intact after the statoblast is set free. The statoblast does not float until it is dry, and the float is similar in size on both valves. In Group II, a mature statoblast is covered by a softgelatinous basophilic layer, which decays after the statoblast is released. The statoblast floats without drying, and the float is better developed on the dorsal valve than on the ventral. Moreover, in the members of Group II, large yolk granules are first formed, followed by much smaller yolk granules. When their statoblasts are treated with KOH, the shell is separated completely into two valves. These characters are common to many lower phylactolaemates. By contrast, in L. carteri and P. gelatinosa, the yolk granules are uniformly small and the capsule proper resists KOH treatment. On these points, L. crystallinus is somewhat different from these two species, suggesting its primitive nature.     

Fusion of ancestrulae germinated from statoblasts in plumatellid freshwater bryozoans

Upon germination of a statoblast, the shell is split into two valves; a mucous pad which represents the basal portion of the body wall of the incipient zooid or ancestrula then appears from between the valves; lastly, a tiny polypide evaginates at the opposite site. When two or more contiguously located statoblasts (floatoblasts or sessoblasts) of the same species germinate simultaneously, their mucous pads often come into contact with each other. The walls of the mucous pads then disappear in the contact areas, thus uniting the coeloms of the ancestrulae. This type of fusion between mucous pads of statoblast-derived ancestrulae was ascertained in Plumatella emarginata, P. repens, P. casmiana, and Hyalinella punctata. The fusion is clearly species specific, and shows no clone specificity or allogeneic recognition. The fusibility test reported here seems to be a useful method for the examination of conspecificity in plumatellid bryozoans.     

A benign approach to the preparation of freshwater bryozoan statoblasts for scanning electron microscopy (SEM) imaging

Abstract

Several different species of freshwater Bryozoa, belonging to the genera Plumatella, Rumarcanella and Fredericella, were detected within the Northern Mallee Pipeline (NMP) system in Victoria, Australia, that required definitive identification. These organisms produce asexual buds called statoblasts, with valves composed of sclerotised chitin that bear minute micro-ornamentations of considerable taxonomical significance. Imaging and analysis of these distinctive micro-ornamentations using scanning electron microscopy (SEM) is often employed for species identification. Meticulous preparation of statoblast samples is therefore required that necessitates the removal of adhering debris, dehydration and drying—whilst mitigating specimen damage and distortion. This technical note describes an approach whereby each of these three steps have been individually designed to be as benign as possible, using mild detergent/sonication to remove debris, a gradual and gentle dehydration procedure using ethanol, and critical point drying. For the overall process, these methods are chosen to optimise control and to minimise the use of harsh and hazardous chemicals.     

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.     

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     

Variation in reproductive characteristics of colonies of the freshwater bryozoan Cristatella mucedo

1. We studied relationships between different forms of reproduction and local variation in the reproductive state of colonies of a common freshwater bryozoan Cristatella mucedo Cuvier (Bryozoa: Phylactolaemata). Four sampling locations in central Finland, including both lotic and lentic habitats, were sampled by scuba diving. The traits studied were occurrence of sexual larvae, colony size, number of resting stages (statoblasts) and number of zooids. 2. While 76.9% of the sampled colonies carried statoblasts at the time of sampling, only 4.5% of the colonies were brooding sexual larvae. Most of the colonies were relatively small with a mean colony size of 16.6 mm. 3. In colonies brooding larvae, the number of statoblasts was positively correlated with the number of larvae. We did not detect a colony size threshold for sexual reproduction or formation of statoblasts. Colonies carrying statoblasts had a lower number of zooids per unit dry weight. 4. We found significant variation in reproductive traits of colonies among the sampling locations, and among lotic and lentic habitats. The observed phenotypic differences may reflect broadly distributed, phenotypically plastic clones. Differences in clonal composition of local populations cannot, however, be ruled out and factors that may provide clonal diversity are discussed.     

Asajirella gelatinosa in Panama: a bryozoan range extension in the Western Hemisphere (Ectoprocta: Phylactolaemata)

Asajirella gelatinosa, a freshwater bryozoan known previously only from eastern Asia is now reported in the Republic of Panama. Colonies were collected during April 1992 in the broad reaches of the lower Río Chagres as it enters Gatun Lake, part of the Panama Canal system. A subsequent collection in 1998 included large colonies and numerous statoblasts from a region upstream of this site in the impounded Lago Alajuela (Madden Lake). Colonies and statoblasts resemble those from Japan in almost every detail. Human transport of statoblasts is a likely cause of this disjunct distribution. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cryptic invaders: nonindigenous and cryptogenic freshwater Bryozoa and Entoprocta in the St. Lawrence River

The distributions of most cosmopolitan invertebrate species are assumed to result from natural processes. Cryptic invertebrates with obscure biogeographic origins are often considered native by default, resulting in potentially severe underestimation of the extent of human-assisted invasions. This problem is exemplified by freshwater Bryozoa (Ectoprocta) and Entoprocta—small and widely distributed invertebrates commonly found in lakes and rivers. A benthic survey of a thermally modified section of the St. Lawrence River revealed the presence of two nonindigenous bryozoans: Carter’s moss animal Lophopodella carteri (Hyatt) and the crystal moss animal Lophopus crystallinus Pallas. Also discovered was a cryptogenic entoproct, the goblet worm Urnatella gracilis Leidy. These species were collected as statoblasts and (in the case of U. gracilis) colonial fragments downstream of the Gentilly-2 nuclear power plant at Bécancoeur, Quebec. Local densities of both U. gracilis and L. carteri increased by an order of magnitude at sites closer to the power plant. The occurrence of Lophopus crystallinus statoblasts in St. Lawrence River sediments is the first documented physical evidence of the species in North America. Contrary to the presumed natural Holarctic distribution of L. crystallinus, our literature review found that published historical records of L. crystallinus in the United States are erroneous or unsubstantiated. We propose that L. crystallinus is a western Palearctic species recently introduced to the St. Lawrence River, most likely as statoblasts discharged with ballast water from transoceanic ships.     

Bryozoan populations reflect nutrient enrichment and productivity gradients in rivers

1. The hypothesis that nutrient enrichment will affect bryozoan abundance was tested using two complementary investigations; a field‐based method determining bryozoan abundance in 20 rivers of different nutrient concentrations by deploying statoblast (dormant propagule) traps and an experimental laboratory microcosm study measuring bryozoan growth and mortality. These two methods confirmed independently that increased nutrient concentrations in water promote increases in the biomass of freshwater bryozoans. 2. Statoblasts of the genus Plumatella were recorded in all rivers, regardless of nutrient concentrations, demonstrating that freshwater bryozoans are widespread. Concentrations of Plumatella statoblasts were high in rivers with high nutrient concentrations relative to those with low to moderate nutrient concentrations. Regression analyses indicated that phosphorus concentrations, in particular, significantly influenced statoblast concentrations. 3. Concentrations of Lophopus crystallinus statoblasts were also higher in sites characterised by high nutrient concentrations. Logistic regression analysis revealed that the presence of L. crystallinus statoblasts was significantly associated with decreasing altitude and increasing phosphorus concentrations. This apparently rare species was found in nine rivers (out of 20), seven of which were new sites for L. crystallinus. 4. Growth rates of Fredericella sultana in laboratory microcosms increased with increasing nutrient concentration and high mortality rates were associated with low nutrient concentrations. 5. Our results indicate that bryozoans respond to increasing nutrient concentrations by increased growth, resulting in higher biomasses in enriched waters. We also found that an important component of bryozoan diets can derive from food items lacking chlorophyll a. Finally, bryozoans may be used as independent proxies for inferring trophic conditions, a feature that may be especially valuable in reconstructing historical environments by assessing the abundance of statoblasts in sediment cores.     

Annual net production and life span of floating leaves in Nymphaea tetragona Georgi: a comparison with other floating-leaved macrophytes

Fredericella sultana (Blumenbach, 1779) has long been considered one of the few freshwater bryozoan species with a truly cosmopolitan distribution. However, chromosome spreads from European material show 2n = 16 compared to 2n = 14 in North American specimens. In laboratory rearing the two forms are morphologically indistinguishable except for the surface texture of their statoblasts. Smooth statoblasts of European colonies match early illustrations of the species, while the densely pitted statoblasts of the North American form resemble those of F. indica Annandale 1909. On the basis of these observations we tentatively designate the North American F. sultana as F. indica. The only known American species with smooth statoblasts is F. australiensis Goddard 1909, in which the 2n = 16 karyotype is similar or identical to European F. sultana; however, despite this karyotypic similarity the two species retain their distinguishing morphology when reared together in the laboratory. Two enzymes from a single specimen of European F. sultana were electrophoretically distinct from the corresponding enzymes present in samples of both F. australiensis and North American F. sultana. Four phosphoglucose isomerase alleles were present in North American F. aultana from four geographically separated collection sites, although only one genotype for this locus was observed in material from any one site. These genetic findings are consistent with a relatively short-range dispersal potential in this species as compared to Plumatella species.     

NOR-Chromosome morphology and evidence for rDNA selection in phylactolaemates

Silver-stained chromosome spreads from the phylactolaemate ectoprocts Fredericella indica, Fredericella browni, Plumatella repens, Pectinatella magnifica and Asajirella gelatinosa show that each of these species has a single pair of NORs. In the first four species each NOR is immediately adjacent to the centromere of a large metacentric chromosome. Giemsa-stained preparations from Cristatella mucedo show centromeric gapping in a similar large metacentric, indicating the same type of NOR-chromosome. In Asajirella gelatinosa the two NORs are on a pair of smaller chromosomes, and are not immediately adjacent to a centromere. A close NOR-centromere association would inhibit meiotic crossing over in the NOR region. Since in phylactolaemates reproduction is primarily asexual, often by formation of statoblasts, there is potential intraclonal selection for favorable ribosomal DNA (rDNA) mutations, including from simultaneous germination of otherwise genetically identical statoblasts within a limited area and competition between cell lines within statoblasts. Inhibition of meiotic crossing over within the NOR would maintain its integrity during periods when intraclonal selection would not be occurring. This suggests that a more rapid rate of rDNA evolution is possible in phylactolaemates than in most other organisms, and is consistent with a derivation of this group from the morphologically similar phoronids, despite phylogenetic interpretations of rDNA sequence data that suggest the two groups are not closely related.     

Chromosomal heteromorphism in a Japanese population of Pectinatella gelatinosa and karyotypic comparison with some other phylactolaemate bryozoans

Five different karyotypes were present in germinating statoblasts obtained from a sampling of a predominately or exclusively asexually reproducing population of Pectinatella gelatinosa from Lake Tatara-numa, Japan. Karyotypes were consistent in having 18 chromosomes, a unique telocentric, a pair of minute metacentrics, and at least one prominently secondarily constricted element. In a number of cases, a sampling of statoblasts from a single large colony yielded more than one karyotype, perhaps because there had been fusion of smaller colonies with different karyotypes. Some individual statoblasts may have been chimeric, but this was not conclusively demonstrated. The possibility of asexually-maintained permanent chimerism in phylactolacmates involving karyotypically distinct ectodermal and peritoneal cell lines is discussed. Karyotypically, P. gelatinosa is more similar to Lophopodella carteri than to Pectinatella magnifica.     

Dormant stages in freshwater bryozoans--an adaptation to transcend environmental constraints

Freshwater invertebrates often disperse between discrete habitat patches via the production of dormant propagules. Being dispersed passively by animal vectors or wind, certain adaptations for exposures to terrestrial and aerial conditions like desiccation and freezing are required. In the present study, we investigate the mechanisms of survival and physiological adaptations due to desiccation and low temperatures in the statoblasts of two populations of the freshwater bryozoan Cristatella mucedo. Our results show that both sessoblasts and floatoblasts tolerate almost complete desiccation and subzero temperatures. Trehalose, a non-reducing disaccharide which has been related to desiccation tolerance, was detected by amperometric chromatography. However, due to the low concentrations found, it is unlikely that trehalose is playing a major part in desiccation tolerance of bryozoan statoblasts. Vitrification is assumed to be important in the survival of desiccation tolerant organisms. Differential scanning calorimetry revealed thermal transitions (T_g onset around 70 °C) in desiccated statoblasts, indicating that a vitreous matrix is present. During the exposure to subzero temperatures, freeze tolerance of statoblasts was confirmed by the detection of internal ice formation, which took place at a crystallisation temperature of between −6 °C and −12 °C.     

Plumatella mukaii,a new phylactolaemate bryozoan from Asia and South America

A new freshwater bryozoan species, Plumatella mukaii, is recognized from eastern Asia and western South America. Colonies and statoblasts both bear a resemblance to P. emarginata, and have often been confused with that species, especially in Japanese studies. Floatoblasts and sessoblasts are enclosed within a tough, wrinkled, membrane which resists removal by mechanical means. Floatoblasts are generally smaller than those of P. emarginata, but display unusually high variability in their overall dimensions. The species has been reported from both lentic and lotic habitats. In Asia, the range includes Japan, Korea, China, India and Indonesia. It has most recently also been found at several sites in Chile. The recognition of P. mukaii narrows the previously reported range of P. emarginata and invites a re-inspection of that species worldwide.     

Experimental cultivation of the invasive freshwater bryozoan <Emphasis Type="Italic">Pectinatella magnifica</Emphasis>

The freshwater bryozoan Pectinatella magnifica (Leidy, 1851) is an invasive species in many countries all over the world. Although native to North America, it has been found in many countries of Europe and Asia. Pectinatella magnifica forms the largest colonial masses from all recently known bryozoan species. Culturing this organism in an aquarium has never been achieved for more than few days so far. Colonies from laboratory culture are important for various studies on its biology and life cycle of this species in experimental conditions. Young colonies successfully hatched from germinating statoblasts of P. magnifica in the laboratory and were maintained over eight weeks. Moreover, this was the first time when the compound colonies of this species were carried from its natural habitat to the laboratory, into a special aquarium system, and kept alive for more than three weeks. In both experiments the physicochemical parameters of the water (temperature, concentration of dissolved oxygen, electrolytic conductivity and pH) and changes in weight of compound colonies of P. magnifica in laboratory conditions were checked. The results found in this study are essential for understanding the invasiveness of this species and identifying methods for elimination of its ecological risks because these are closely resembling those of other invasive species.     

Interpretation of bryozoan microfossils in lacustrine sediment cores

Bryozoan statoblasts were isolated from the surficial sediments of 30 Florida lakes representing a gradient of trophic conditions. A prerequisite for lake selection was a good data base for water chemistry that was collected within the past 5–10 years. A novel technique of employing 210-Pb as a dilution tracer was used to estimate sedimentation rates for each lake. The net annual accumulation rate for bryozoan statoblasts was calculated for each lake and correlated with twelve physical, chemical, and biological variables. The results suggest that bryozoan distributions are strongly controlled by the extent of the littoral zone and phytoplankton biomass. It appears that bryozoan abundance in excessively weeded lakes may be limited by phytoplankton in spite of the richly developed habitat. Separation of the importance of food requirements from habitat requirements was not possible from this initial survey.     

Shallow lake sediments provide evidence for metapopulation dynamics: a pilot study

The aim of this pilot study is to test the hypotheses that sediment cores can provide evidence for metapopulation dynamics and that these can be linked with site characteristics. We focus on temporal patterns of incidence and abundance of overwintering stages (statoblasts) produced by the freshwater bryozoan Cristatella mucedo, an organism characterised by a metapopulation ecology, in sediment cores retrieved from 18 UK lakes. Runs and goodness-of-fit tests provided evidence for population instability, periods of low abundance and absence, and of asynchrony—all signatures of metapopulation dynamics. Further hypothesis testing indicated that extinction risk is greater in more isolated sites and in sites of smaller size. Absence of statoblasts from the top sections of spatially separated, replicate cores provided independent evidence for extinction in one site. Our study demonstrates how the abundances of sedimentary-bound propagules may be analysed initially for metapopulation dynamics and subsequently how this may lead to working hypotheses regarding the drivers of such dynamics. The sediment archive represents a unique historical record whose potential for characterising metapopulation dynamics has previously been untapped but is broadly relevant for understanding the population biology of freshwater organisms.     

The shell of invertebrate and plant propagules: selective permeability and barriers

In the experiments with seeds of Ericaceae, gemmuls of Spongia, statoblasts of Bryozoa, resting eggs and ephippia of Crustacea the hypothesis on selective permeability of propagule shells was tested. It was shown that the propagules are activated and develop until the moment of destruction of shell in the solutions of phenols, formaldehyde, CuSO4, K2Cr2O7, HgCl2, Ag2SO4, AgNO3, CoCl2 and NiSO4. It means that propagule shell is impermeable for these poisons but permeable for water and oxygen. The effectiveness of barrier can be various in different species but in some cases it is very high. The development of resistance to poisons of anthropogenic origin that were not familiar to organism earlier can be considered as an example of the rule (Rasnitzyn, 1971) according which relation between environmental variability and power of regulator is shifted to the priority of regulator. Selectively permeable shell for many organisms is a common adaptation that allows colonization of land and continental waters.