Density-dependent processes in cohorts of Tubifex tubifex,with special emphasis on the control of fecundity

Laboratory cohort cultures of the tubificid Tubifex tubifex with different initial densities were carried out at 20° C with the condition of unlimited food. The main results were: 1) Intracocoon mortality was 37% of the laid eggs (observation of 689 eggs); 2) The principal bionomic parameters (generation time, r, R₀) appeared to be density dependent; 3) Recruitment was regulated through the percentage of worms that actually attained the ovigerous stage, specific fecundity, and the duration of the egg laying stage, which appeared to be inversely correlated with density.     

Population analysis in mass cultures of Tubifex tubifex

Mass cultures of Tubifex tubifex from Lake Orta were kept in the lab at 20 °C starting with different initial densities (10, 45, 86, 161 ind · jar^–1), with the aim of evaluating the effect of density on population numbers and on population parameters. The results show that density mainly controls fecundity, growth, maturation and ovigeration rates. Growth rates and mean number of eggs laid/ovigerous (R₀) are inversely related to density, but generation time appears to be directly related. Very low or very high initial densities display, at different times of the culture history, efficient density controls. Intermediate N₀ seem to bring about situations of bad control, easily leading to numerical overshoots. A possible implementation of a population dynamics model (Bonomi & Di Cola, 1980), originally used for laboratory observations on T. tubifex cohorts was considered, including some density control functions derived from our observations on cohort and mass cultures in some transfer rates and in fecundity rate.     

A comparison of two tubificid oligochaete species as candidates for sublethal bioassay tests relevant to subtropical and tropical regions

Two tubificid oligochaetes, Branchiura sowerbyi and Tubifex tubifex, were evaluated as potential test organisms for sediment bioassays. We attempt to reproduce the sediment bioassay proposed by Reynoldson et al. (1991) using his strain of Tubifex tubifex and his reference sediment and to compare this technique using Branchiura sowerbyi. This species was chosen because it is more common and dominant in tropical and subtropical environments than Tubifex tubifex. Data on survival and reproduction were obtained for both species, and growth estimates were obtained for B. sowerbyi. The sublethal bioassay with T. tubifex confirmed earlier estimates of a test duration of 4 weeks at 22.5 °C. B. sowerbyi cultures can produce usable estimates in 21 days at 30 °C.     

Assimilation by Aquatic Oligochaeta

Aquatic oligochaetes selectively ingest material from sediments given as food. Determinations of the quantity of organic material presented, left in the residue, and voided as feces enabled estimation of assimilation. These figures were not significantly different from those obtained by summing growth as observed in the experiment with respiration determined independently. Short term experiments yield estimates of assimilation of 77 to 181 cals/g dry wt/day, long term experiments yield an estimate of 232 cals/g dry wt/day, both for mixed populations of Tubifex tubifex and Limnodrilus hoffmeisteri.     

Preliminary investigations on the utilization of tubificid worms by postlarvae of Macrobrachium lanchesteri (de Man)

Postlarvae of Macrobrachium lanchesteri were stocked in confined waters. The larvae were fed on an ad libitum food of Tubifex tubifex worms for 30 days and their rearing has been established. 1 Kg of initially stocked postlarvae need 4.5 Kg of live T. tubifex to produce 0.537 Kg of new flesh. The worms were found to increase the crude protein content of larval flesh by 17% of the dry matter.     

Countering morphological ambiguities: development of a PCR assay to assist the identification of <Emphasis Type="Italic">Tubifex tubifex</Emphasis> oligochaetes

The freshwater oligochaete, Tubifex tubifex, is a common resident of organic-rich sediments worldwide. Although it is a familiar species to fish enthusiasts, toxicologists and parasitologists, T. tubifex often confounds definitive identification due to the similarity that immature specimens bear to several other common oligochaetes, and given the degree of plasticity of key morphological characters due to environmental conditions and/or age of specimen. To solve this identity crisis, we used a polymerase chain reaction based molecular approach and developed a T. tubifex specific assay with primers that amplify a 192 bp fragment of the internal transcribed spacer region 1 ribosomal DNA. We tested these primers on four T. tubifex mitochondrial genotypes, and on other oligochaete species from nine genera. The primers amplified all specimens identified morphologically asT. tubifex. They did not amplify any other species, including morphologically similar worms possessing hair chaetae (Dero digitata, Ilyodrilus templetoni, Tubifex ignotus or Rhyacodrilus spp.) or other oligochaetes often found with T. tubifex (Lumbriculus variegates, Limnodrilus hoffmeisteri, Stylodrilus heringianus or Trichodrilus sp.). This technique should remove the uncertainties all too often associated with identification of T. tubifex.     

A comparison of reproduction, growth and acute toxicity in two populations of Tubifex tubifex (Müller,1774) from the North American Great Lakes and Northern Spain

Reproduction in Tubifex tubifex is being used as part of a suite of indicators of sediment toxicity in Canada and Spain, and reproduction of T. tubifex is being considered as a component of sediment objectives for environmental regulation and clean-up in the Canadian Great Lakes. The data being used to set these reproductive targets have been developed from a single culture of T. tubifex from Lake Erie. The plasticity of this particular species is well known and before it can be adopted widely as a test organism it is necessary to determine whether a single culture source should be used or if cultures derived from different populations respond similarly. A series of experiments with two cultures, one from Lake Erie the second from a small mountain stream in Northern Spain have shown that the Spanish worms appear to produce fewer cocoons per adult (mean 8.6 S.D. 1.0) than those from Lake Erie (mean 10.4 S.D. 0.3) at 22.5 °C, a standard test temperature. The number of young produced per adult by the Spanish culture is also lower (mean 19.0 S.D. 4.6) than the L. Erie population (mean 30.6 S.D. 2.3), however, the Spanish population has higher reproductions rates at a lower temperature. The Spanish worms also have lower and more variable growth rates than the Canadian population. There also appear to be slight differences in the sensitivities to toxicants, with the Canadian worms having higher LC50s for copper, chromium and cadmium. While there are differences in the responses in the two cultures these are not considered to be sufficient to invalidate the use of either population in a standard bioassay protocol as long as appropriate calibration and validation are undertaken.     

Community metabolism on rocky-shore assemblages in a mesocosm: A. Fluctuations in production,respiration, chlorophyll a content and C:N ratios of grazed and non-grazed assemblages

Studies were undertaken with the aim of developing a standardized method for assessing environmental pollution in sediments by utilization of life-history data of freshwater tubificids. Similar bioassay methods have long been used for Daphnia magna, species of Ceriodaphnia and Nitocra, etc. in accordance with guidelines from the International Organization for Standardization (ISO). Tubifex tubifex was found to be the most likely candidate for such bioassays, since the species is readily kept in culture and reproduces more or less consistantly.The culturing method is slightly modified from Kosiorek (1974). This paper provides an example of the particular sensitivity of this kind of bioassay method in the detection of heavy metal contamination of lake sediments. Sediments from the oligotrophic Lake Runn were considered suitable for the purpose, since the lake receives waste water from a major mining industry in Sweden. Metal analyses of the sediments had revealed the agents likely to be causing the decreased biological activity measured in the lake; rough amplitudes for mercury: 800–3600 ng · g^-1 dw, copper: 800–1800 g · g^-1 dw, zinc: 3.3 – 8.1 mg · g g^-1 dw have been estimated for surficial sediments.Young tubificids exposed to Lake Runn sediments did not grow much and died off within a short period of time. No reproduction occurred. Sediments from Lake Runn, when mixed with sediments from the eutrophic Lake Hjälmaren, made reproduction of T. tubifex occur only in mixtures containing less than 50% L. Runn sediments. The growth rate, reproductive success and the very timing of consecutive reproductive events of cohort individuals were found to be highly indicative of toxic effects. When additional food sources were available, however, these effects were largely masked. Therefore, extra food rations were excluded from the original method.     

Cohort cultures of Tubifex tubifex forms

By means of cohort cultures at different temperatures, the life cycles of two sympatric forms of T. tubifex have been compared: the normal T. tubifex tubifex and the blanchardi forms. Although the two forms have certain basic similarities, the blanchardi form tolerated low temperatures less well, requiring a longer time to develop to maturity and producing fewer ova. The threshold temperature for development appeared to be around 8 °C, notably higher than for the normal form around 0 °C. Moreover, for the blanchardi form, resistance was lower and mortality higher for embryos and throughout the life cycle. Cross-breeding tests were carried out, along with cultures of individuals serving as controls for partheno-genetic activity. In cross-breeding, pairs produced progeny similar to the normal form only, most likely derived not from eggs fertilized by the partner, but from parthenogenetic eggs. As a matter of fact, only the normal form was able to reproduce through parthenogenesis. Thus, the two forms have remarkable differences, indicating genetic differences, and they may be considered not as forms, but rather as distinct species.     

Anaerobic digestion of cellulose by pure and mixed bacterial cultures

Summary By enrichment technique, nine anaerobic mixed bacterial cultures were isolated, five of which showed stable cellulolysis. All cultures fermented cellulose and produced different fermentative products. Mixed culture BOC 25 yielded major levels of acetate and ethanol (39.6 and 12.0 mmol/l, respectively) and minor levels of propionate (2.5 mmol/l) and digested filter paper cellulose to the extent of 32.5% w/v. BOC 25 digested cellulosic and lignocellulosic substrates and produced filter paper cellulase, carboxymethyl cellulase, Avicelase and -glucosidase. Strain DC 25, a cellulolyticClostridium was purified from one of the mixed cultures. The fermentation products of DC 25 from cellulose, cellobiose or glucose were ethanol, acetate, formate, H₂ and CO₂.     

Cyst formation of Tubifex tubifex (Müller) — an adaptation to survive food deficiency and drought

Adult individuals of Tubifex tubifex (Müller) (Oligochaeta, Tubificidae) could be induced to form cysts in the laboratory. Hiding in cysts, Tubifex tubifex survived a five-month drought period in the field. Encysted Tubifex tubifex had a lower super-cooling-point than mobile individuals. Food shortage is believed to be the main factor in cyst formation.     

Endoglucanase Activity of Compost-Dwelling Fungus Paecilomyces inflatus is Stimulated by Humic Acids and Other Low Molecular Mass Aromatics

Summary Paecilomyces inflatus isolated from municipal waste compost was found to have cellulolytic activity in several solid and liquid media. This study was done to reveal the multifarious effects of municipal waste compost on endoglucanase activity of P. inflatus. The highest enzyme activities under the conditions of solid-state fermentation were measured in authentic compost samples compared with wood, straw and bran substrates. In surface liquid cultures glucose, cellobiose, xylan, Avicel cellulose, carboxymethylcellulose (CM-cellulose), starch and citrus pectin were used as carbon sources. All carbon sources supported the growth of P. inflatus. However, only CM-cellulose, cellobiose and pectin noticeably stimulated endoglucanase (EG) activity. Further stimulation of EG activity was obtained in cultures containing 1% CM-cellulose as a carbon source by supplementation with low-molecular mass aromatic compounds vanillin, veratric acid and benzoic acid, and with soil humic acid (SHA). SHA and veratric acid were found to be the most efficient elicitors of the cellulolytic activity. P. inflatus was able to utilize nitrate and ammonium as pure nitrogen sources in media containing cellulose.     

Growth of Tubifex tubifex Müller (Oligochaeta,Tubificidae) under Various Trophic Conditions

Growth of Tubifex tubifex on highly caloric activated sludge and poor in organic matter natural silt was experimentally studied. The relationship between oxygen consumption rate, weight, and caloric value of T. tubifex body in culture on activated sludge was determined to calculate the growth efficiency (K₂ = P/P + R). The growth rate has been determined, and models of T. tubifex growth are given under various trophic conditions conforming to results of the experiment. Somatic growth, generative growth and life span of this species were compared on the basis of the experiments and literature. Acceleration of growth and prolongation of life under conditions of caloric food and delay of these processes in silt poor in organic matter have been established and measured. T. tubifex was found to have advantages as compared to other tubificid species in response to increasing amounts of nutrient food in waters polluted by organic matter and, therefore, to produce abundant populations under these conditions.     

Expression pattern of annelid <Emphasis Type="Italic">Zic</Emphasis> in embryonic development of the oligochaete <Emphasis Type="Italic">Tubifex tubifex</Emphasis>

Embryonic expression of a Zic homologue (Ttu-Zic) was examined in the oligochaete annelid Tubifex tubifex. The body plan of T. tubifex is characterized by obvious segmentation in the ectoderm and mesoderm. Ttu-Zic expression is detected in the mesodermal germ band and a subset of micromere descendants. Ttu-Zic is transiently expressed in primary m-blast cells (i.e., founder cells of mesodermal segments) as early as the time of their birth from M teloblasts. During its development, each mesodermal segment experiences two additional phases of Ttu-Zic expression. Ttu-Zic expression in micromere descendants is seen on the anterior surfaces of embryos undergoing teloblastogenesis; subsequently, these cells proliferate to form bilateral clusters, which then become internalized. Finally, clusters of Ttu-Zic-expressing cells are found in the center of the prostomium, corresponding to the cerebral ganglion. The Ttu-Zic expression profile in the early embryogenesis of T. tubifex may be homologous to those of evolutionarily distant animals.     

Monoxenic culture of the anaerobic ciliate Trimyema compressum Lackey

Sapropelic ciliates from anoxic mud samples were enriched and cultivated in monoculture together with natural food bacteria growing on cellulose. The ciliates lacked cytochrome oxidase and contained bluish fluorescent endosymbionts. One of the anaerobic ciliates, Trimyema compressum, contained methanogenic bacteria as was shown by methane formation. During continued cultivation, T. compressum gradually lost its endosymbionts. With SEM microscopy no episymbiotic bacteria could be detected.From enrichment cultures of T. compressum, anaerobic bacteria were isolated in pure culture. One of the strains, a Bacteroides spec., proved capable of serving as food bacteria, thus allowing establishment of monoxenic T. compressum cultures. These cultures exhibited a requirement for sterols as growth factors. The doubling time of this ciliate was 13 h at 28°C. The highest yield obtained was 2100 cells/ml.Dedicated to Holger W. Jannasch on the occasion of his 60th birthday     

Primary simple assays of cellulose-degrading fungi

Some 25 fungi, including at least 14 basidiomycetes, one ascomycete, and five anamorphic fungi were evaluated for their cellulose-degrading abilities in Difco potato dextrose broth or Difco malt extract broth cultures with cellulosic substrates (e.g., filter paper) in plastic Petri dishes. Among them, Peniophora sp. 06-13 and Phlebia sp. 99-335 reduced the dry weights of the whole cultures with these substrates more than the dry weights of the respective original substrates after 30 days of culture, showing definite cellulose degradation. In the cultures with more than 10 test fungi including Pycnoporus coccineus 84-117, such weight losses did not occur. This assay technique for the primary screening for cellulose degrading fungi is simple, inexpensive, reproducible and accurate.     

Cell suspension cultures of Populus tremula × P. tremuloides exhibit a high level of cellulose synthase gene expression that coincides with increased in vitro cellulose synthase activity

Summary. Compared to wood, cell suspension cultures provide convenient model systems to study many different cellular processes in plants. Here we have established cell suspension cultures of Populus tremula L. × P. tremuloides Michx. and characterized them by determining the enzymatic activities and/or mRNA expression levels of selected cell wall-specific proteins at the different stages of growth. While enzymes and proteins typically associated with primary cell wall synthesis and expansion were detected in the exponential growth phase of the cultures, the late stationary phase showed high expression of the secondary-cell-wall-associated cellulose synthase genes. Interestingly, detergent extracts of membranes from aging cell suspension cultures exhibited high levels of in vitro cellulose synthesis. The estimated ratio of cellulose to callose was as high as 50 : 50, as opposed to the ratio of 30 : 70 so far achieved with membrane preparations extracted from other systems. The increased cellulose synthase activity was also evidenced by higher levels of Calcofluor white binding in the cell material from the stationary-phase cultures. The ease of handling cell suspension cultures and the improved capacity for in vitro cellulose synthesis suggest that these cultures offer a new basis for studying the mechanism of cellulose biosynthesis. Correspondence and reprints: School of Biotechnology, Royal Institute of Technology, AlbaNova University Centre, 106 91 Stockholm, Sweden. Present address: Department of Biotechnology, Beijing Forestry University, Beijing, People’s Republic of China     

Primary production in ten reservoirs in southern Brazil

The growth and phosphatase activity during phosphorus starvation of cultures of Nodularia spumigera Mertens were examined. Stationary phase was reached much sooner in phosphorus-deficient cultures than in phosphorus-sufficient cultures; the growth rate did not change. Phosphatase activities were greatly increased in stationary phase. Diurnal patterns were established for phosphorus-sufficient cultures, but they were not light related. In phosphorus-deficient cultures, an increase in phosphatase activities over a 24 h period was superimposed on a diurnal pattern. Both phosphorus and nitrogen additions lowered the relative phosphatase activities in long term studies, but the effect of phosphorus was much more pronounced. In short term studies, phosphorus appeared to cause an immediate decrease in phosphatase activity, but did not affect phosphatase activity after that for up to 24 h. Nitrogen did not have any short term effect on phosphatase. Phosphatase activity was correlated with changes in the proportion of TCA-insoluble phosphorus (polyphosphates).     

Some characteristics of genetic variants of Tubifex tubifex (Müller, 1774) (Oligochaeta: Tubificidae) in laboratory cultures

Genetic variants of the oligochaete Tubifex tubifex were identified with enzyme electrophoresis and subsequently reared in laboratory cultures. Three types are abundant in field populations. Individuals that show homozygotic bands of glucosephosphate-isomerase (GPI) 22 together with isocitrate-dehydrogenase (IDH) 35 were labeled Type A. Type B is characterized by GPI 23 together with IDH 11 and Type C is characterized by GPI 11 with either IDH 34 or IDH 33. Initial results on freshweights of adults and cocoon production revealed differences between the two main types, A and B. In the same period, Type B reached higher weights and produced five times as many cocoons as Type A, whereas number of eggs per cocoon were not different between these Types. Type B also had the lowest mortality in 16-week experiments with changing temperatures.     

Degradation of food compounds and growth response on different food quality by the anaerobic ciliate Trimyema compressum

The biochemical composition of two food bacteria was examined on which monoxenic cultures of Trimyema compressum grew with different yields. The food bacteria were the saccharolytic fermenting bacterium Bacteroides WoCb15 and the purple nonsulfur bacterium Rubrivivax gelatinosus. Differences in composition of bacterial biomass concerned mainly the carbohydrate content. By different culture conditions for R. gelationsus and pasteurization of carbohydrate-rich cells, we were able to feed the ciliate with food mixtures of different carbohydrate content. Dry mass yields of the ciliate reached a maximum with mixtures of 80% carbohydrate-rich pasteurized cells plus 20% carbohydratepoor living cells. In the absence of degradable carbohydrate, energy metabolism depended on protein as substrate. Utilization of protein was incomplete, large amounts were converted into soluble compounds that accumulated in the culture medium. The ciliate consumed storage carbohydrate of living or pasteurized food bacteria equally well, while growth with short generation times was still dependent on a certain percentage of living bacteria as source of native protein. Lipids, nucleic acids and denatured proteins were not degradable by the ciliate. Consequences for the fermentative metabolism of Trimyema compressum are discussed.