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.     

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.     

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.     

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.     

Convergent evolution of the lateral line system in Apogonidae (Teleostei: Percomorpha) determined from innervation

The lateral line system and its innervation were examined in two species of the family Apogonidae (Cercamia eremia [Apogoninae] and Pseudamia gelatinosa [Pseudamiinae]). Both species were characterized by numerous superficial neuromasts (SNs; total 2,717 in C. eremia; 9,650 in P. gelatinosa), including rows on the dorsal and ventral halves of the trunk, associated with one (in C. eremia) and three (in P. gelatinosa) reduced trunk canals. The pattern of SN innervation clearly demonstrated that the overall pattern of SN distribution had evolved convergently in the two species. In C. eremia, SN rows over the entire trunk were innervated by elongated branches of the dorsal longitudinal collector nerve (DLCN) anteriorly and lateral ramus posteriorly. In P. gelatinosa, the innervation pattern of the DLCN was mirrored on the ventral half of the trunk (ventral longitudinal collector nerve: VLCN). Elongated branches of the DLCN and VLCN innervated SN rows on the dorsal and ventral halves of the trunk, respectively. The reduced trunk canal(s) apparently had no direct relationship with the increase of SNs, because these branches originated deep to the lateral line scales, none innervating canal neuromast (CN) homologues on the surface of the scales. In P. gelatinosa, a CN (or an SN row: CN homologue) occurred on every other one of their small lateral line scales, while congeners (P. hayashii and P. zonata) had an SN row (CN homologue) on every one of their large lateral line scales.     

Histological and histochemical studies on the formation of statoblasts of a fresh-water bryozoan,Pectinatella gelatinosa

The course of the statoblast formation in Pectinatella gelatinosa was divided into four stages and studied histologically and histochemically. The bottom of the cystigenous cup is a center of cystigenous cell differentiation and the peripheral zone of the inner cystigenous layer turns to the outer cystigenous layer as the cystigenous cup grows. The annulus is formed by migration and transformation of the outer cystigenous cells. During early stages, the yolk cells have an intensely pyroninophilic or RNA-rich cytoplasm. The cytoplasmic pyroninophilia then diminishes as the amount of yolk granules increases. Several kinds of yolk substances occur in the mature statoblast. During statoblast formation glycogen appears first, then glycoprotein and finally neutral unsaturated lipid. Acid phosphatase activity is associated with granular structures in the cytoplasm. In the cystigenous vesicle, acid phosphatase activity is low and confined to the apical extremity of the cell. Histochemically detectable alkaline phosphatase activity is not involved in the formation of the statoblast.     

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.     

Biodegradation of aromatic compounds byRhodopseudomonas gelatinosa

A few photosynthetic bacteria have been isolated from a biological treatment system treating producer gas plant effluent. One of them was identified asRhodopseudomonas gelatinosa on the basis of physiological and morphological characteristics. Biodegradation of aromatic compounds byR. gelatinosa appears not to have been reported in the literature. The culture was found to degrade benzoic acid,p-hydroxybenzoic acid, cinnamic acid, andp-coumaric acid. The doubling time for this culture was found to be 18, 19, 23, and 31 h for benzoic acid, p-hydroxybenzoic acid, cinnamic acid, andp-coumaric acid respectively.     

Ultrastructural studies on the formation of yolk granules in the statoblast of a fresh-water bryozoan,Pectinatella gelatinosa

The formation of protein-carbohydrate yolk in the statoblast of a fresh-water bryozoan, Pectinatella gelatinosa, was studied by electron microscopy. Two types (I and II) of yolk cells were distinguished. The type I yolk cells are mononucleate and comprise a large majority of the yolk cells. The type II yolk cells are small in number; they become multinucleate by fusion of cells at an early stage of vitellogenesis. In both types of yolk cells, electron-dense granules (dense bodies) are formed in Golgi or condensing vacuoles, which are then called yolk granules. For the formation of yolk granules, the following processes are considered: 1. Yolk protein is synthesized in the rough-surfaced endoplasmic reticulum (RER) of the yolk cells. 2. The synthesized protein condenses in the cisternal space of the RER and is packaged into small oval swellings, which are then released from the RER as small vesicles (Golgi vesicles, 300-600 A in diameter). 3. The small vesicles fuse with one another to form condensing vacuoles, or with pre-existing growing yolk granules. 4. In the matrix of the condensing vacuoles or growing yolk granules, electron-dense fibers are fabricated and then arranged in a paracrystalline pattern to form the dense body. 5. After the dense body reaches its full size, excess membrane is removed and eventually the yolk granules come to mature. Toward the end of vitellogenesis of the yolk cells, the cytoplasmic organelles are ingested by autophagosomes derived from multivesicular bodies and disappear.     

Utilization of methanol by rhodospirillaceae

Enrichment culture of organisms growing anaerobically in the light in methanol-bicarbonate medium resulted in isolation of strains of Rhodopseudomonas gelatinosa and Rhodopseudomonas acidophila. The pH optimum for growth on methanol for all strains tested was approximately one unit higher than for growth on carbon sources containing more than one carbon atom. At the appropriate pH, 17 strains of Rhodospirillaceae out of 39 in a culture collection grew anaerobically in the light on methanol-bicarbonate. Rhodopseudomonas acidophila strain 10050 showed the most abundant growth and was studied in more detail. Its growth on methanol was stimulated by yeast extract or vitamin-free casamino acids. The organism grew on methanol-bicarbonate, methanol-formate or formate alone as the sole carbon sources. No growth was observed on methylamine or formaldehyde. In the presence of excess bicarbonate a maximum yield of 98 g cell material from 100 g methanol was obtained. Ribulose diphosphate carboxylase was present in the methanol-bicarbonate-grown organism at six times the specific activity of that in the succinate-grown organism.     

Suitability of Dicloster acuatus MH013435 and Kalenjinia gelatinosa MH012185 for growth in Garcinia cambogia washwater effluent with efficient nutrient removal

Abstract

The present study investigates the suitability of growing Dicloster acuatus MH013435 and Kalenjinia gelatinosa MH012185 in Garcinia cambogia washwater for biomass production and nutrient removal. In recent years, the wastewater effluent treatment with microalgae is gaining importance since it serves multiple purposes including CO₂ sequestration, production of biofuel feedstock and value-added biochemicals in addition to wastewater treatment. Contaminated washwater is generated at a significant quantity during extraction of hydroxycitric acid from G. cambogia and it represents a serious environmental concern when discharged without proper processing. However, this G. cambogia washwater can be used as a low-cost source of nutrients for microalgal growth. The two microalgal species D. acuatus MH013435 and K. gelatinosa MH012185 demonstrated robust growth in washwater and achieved maximum biomass concentration of 0.68 and 0.63?g/L dry weight. The nitrate was removed to an extent of 98.5 and 99.6% whereas chloride removal was 72 and 80.5%, sulfate concentration got reduced by 98 and 98.7% and phosphate got reduced to 95 and 92% for D. acuatus MH013435 and K. gelatinosa MH012185, respectively. The results indicate the suitability of D. acuatus MH013435 and K. gelatinosa MH012185 in treating G. cambogia washwater with efficient nutrient removal.     

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.     

New Structures in Binding Designs of Freshwater Ectoprocta Dormant Bodies (Statoblasts)

With the critical point drying method a scanning electron microscopic study of freshwater Ectoproct dormant structures was made. The suture zones of major phylactolaemate genera were described suggesting methods of their classification. The new structures revealed, throw light on the binding mechanism and early kinetics of germination of these dormant bodies. Median ribs and lateral ribs were described in the suture zones of Plumatella fruticosa, P. casmiana and Pectinatella magnifica. Dormant bodies of Fredericella sultana lack a medial or lateral rib in their suture zone. The fine structure of the polymorphic statoblasts of P. casmiana revealed different binding mechanisms as suited to different seasons. During germination, the internal pressure increases within the statoblast and the well cemented medial rib unfolds, letting its two valves separate. The mechanics of valve separations and subsequent germination were related to their respective environmental conditions favouring their domination.     

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.     

The application of Chlamydomonas cultures as bio-assay test organisms

Summary A method to determine toxicity of individual toxic materials, eventually of sewage and industrial wastes is described. As a test organism the specific pure culture ofChlamydomonas gelatinosa is applied.The growth curves of this organism obtained under optimal conditions are compared with those obtained when different concentrations of matter to be tested had been added. These long term experiments establish not only detrimental or harmful properties of agents investigated, but also the accidental growth stimulative action upon the test organisms.

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Zusammenfassung Es wurde eine neue Methode zur Bestimmung der Toxizität von einzelnen Stoffen oder von Abwässern unbekannter Zusammensetzung beschrieben. Als Test-Organismus wurde eine spezifisch reine (mit Bakterien) Kultur vonChlamydomonas gelatinosa angewendet.Die Wachstumskurven dieses grünen Flagellaten, die unter optimalen Bedingungen gewonnen wurden, wurden mit denen verglichen, die mit Zugabe von verschieden bestimmten Konzentrationen des Giftstoffes erzielt wurden. Diese dreiwöchigen Experimente zeigen nicht nur die lethalen oder schädlichen Einwirkungen des zu untersuchenden Stoffes, sondern auch die eventuelle Wachstumsstimmulation.

     

Evolution of karyotypes and differentiation in 13 Rattus Species

Karyotypes of 13 Rattus species collected in Asia and Oceania were analysed with special interest to karyotype evolution and species differentiation. They were classified into three groups according to their karyotype similarity. Four species (R. annandalei, R. exulans, R. muelleri and R. norvegicus) with 2n=42 and a karyotype similar to some of the polymorphic karyotypes in the Asian black rats (R. rattus) are classified into the first group. Pericentric inversion of some acrocentrics seemed to have caused the differentiation of these species. The other four species (R. bowersii, R. fuscipes, R. leucopus and R. conatus) with similar karyotypes as the above group, but lower chromosome numbers than 2n=42 are classified into the second group. Robertsonian fusion in some acrocentrics observed in the first group are suggested to have caused the development of the species in this group. The remaining four species (R. sabanus, R. canus, R. huang and R. niviventer) with karyotypes markedly different from the above two in having a fewer number of small metacentrics are classified into the third group. They seemed to be more primitive karyotypes than the other Rattus species. By the comparison between the polymorphic karyotypes in the black rat, and karyotypes in its related species it was suggested that the former had occurred as primary events to the differentiation of the latter. Parallelism between the karyotype evolution and the species differentiation was discussed.Contribution No. 874 from the National Institute of Genetics, Japan. Supported by a grant-in-aid from the Ministry of Education of Japan (No. 92159 and 92332).     

Comparison of blood and urine analytes between two karyotypes of owl monkey,Aotus nancymae and Aotus vociferans

Serum and urine analytes were compared between two karyotypes of owl monkey, Aotus nancymae and A. vociferans, to determine if normative clinical pathology data obtained from one karyotype were applicable to the other. Statistically significant differences (P ≤ 0.05) were noted in serum calcium, serum phosphorus, serum sodium, serum potassium, serum urea nitrogen, urine calcium, calcium clearance, and fractional clearance of calcium between the two karyotypes. The results suggest that A. vociferans regulate calcium-phosphorus and electrolyte homeostasis in a manner different from that of A. nancymae.     

Electrophoretic Karyotypes of Candida Yeasts Recurrently Isolated from Single Patients

Candida strains were isolated repeatedly from single patients during recurrent episodes of Candida infection in a hospital, and their electrophoretic karyotypes were analyzed by pulsed-field gel electrophoresis using CHEF system. When only C. albicans (in 6 patients) or C. glabrata (in 1 patient) were recurrently isolated, their karyotypes from each patient were almost identical to one another, suggesting that they carried single type of the yeast. When multiple species were recovered from single patients (6 cases), the karyotypes of the most frequently recovered yeast species were almost identical with respect to each patient. The electrophoretic karyotype analysis has been proved to be useful for epidemiological studies because the method can tell not only the species identification but also the differences among the strains of the same species.