DEVELOPMENT OF PROTOPLASTS FROM HOLDFASTS AND VEGETATIVE THALLI OF MONOSTROMA LATISSIMUM (CHLOROPHYTA, MONOSTROMATACAE) FOR ALGAL SEED STOCK

The aim of this study was to isolate and cultivate the protoplasts of the green alga Monostroma latissimum Wittrock and subsequently induce them to form algal filaments to act as an algal "seed" stock. Protoplasts of the alga were isolated enzymatically with 4% cellulase Onozuka R-10 and 2% Macerozyme R-10. The highest number of protoplasts was obtained on a 50-rpm shaker with 1.2 M of sorbitol after 6 h of incubation, with a yield of 9 × 106 protoplasts·g−1 of fresh thallus (including holdfast). Protoplasts from both holdfasts and erect thalli usually began to form new cell walls within 5 h after isolation and began to divide from day 6 to day 9 in PES medium; cell clusters, filaments, and/or tubular thalli were formed from day 14 to day 18. For algae collected in March, about 60% of protoplasts isolated from vegetative thalli regenerated to form tubular thalli, and about 45% of protoplasts isolated from holdfasts regenerated to form filaments. However, for algae collected in May, about 1% of protoplasts isolated from vegetative thalli developed directly to form tubular thalli, and 59% of protoplasts regenerated to form cell clusters without the ability to differentiate, whereas protoplasts isolated from holdfasts failed to develop. Regenerated filaments were kept in an incubator for more than 3 years at 24° C under the low irradiance of 66μmol photons·m−2·s−1. After this time, they retained the ability to develop to form tubular thalli under irradiance of 166 and 300 μmol photons·m−2·s−1 at 18°–30° C. Subsequently, these tubular thalli can develop to form leafy thalli after being cultivated at high irradiance of 300 μmol photons·m−2·s−1 and at 18°–22° C. Therefore, the filaments could serve as"seed" stock for algal mass culture.     

Development and use of striped bass-specific RFLP probes

We sought to develop nuclear DNA (nDNA) probes which could be used to complement mtDNA and DNA fingerprinting markers in distinguishing striped bass, Morone saxatilis (Walbaum), from discrete spawning systems. Restriction endonuclease-generated single copy, 10–20-kb striped bass nuclear nDNA fragments were cloned into the bacteriophage vector Lambda Dash II and tested in Southern blot analyses for their abilities to reveal population-specific polymorphisms. Three of the I7 nDNA sequences tested exhibited polymorphisms which potentially could be used to delineate striped bass populations. One probe, DSB 22, revealed significant genotypic frequency differences between Gulf of Mexico and Atlantic striped bass and among striped bass representative of some Atlantic systems. These preliminary results suggest that single copy nDNA sequences may provide sufficient polymorphisms to aid in stock identification of species which proved genetically monomorphic using other approaches.     

Preliminary investigation of social interactions and feeding behavior in captive group-housed Tasmanian devils (Sarcophilus Harrisii)

As the number of Tasmanian devils (Sarcophilus harrisii) in captivity increases, an understanding of captive social dynamics and behavior is becoming increasingly important. In the wild, devils are solitary, although sometimes, they congregate to feed on a large carcass. However, it is common to house devils in groups as a form of social enrichment. This study investigated how behavior at feeding time of captive Tasmanian devils varied in groups of different sizes. Observations were made of individually housed devils and devils in groups of two, three, five, and six, when presented with a carcass on which to feed. Total feeding duration ranged from 6.5 to 47.4 minutes per observation period (70 minutes). There was no significant interaction between feeding duration and group size during the experiment. Feeding duration varied daily and depended on carcass size. Social housing of Tasmanian devils enabled them to display dyadic and agonistic behaviors during feeding. Observing behaviors and learning from the outcomes of these interactions can improve husbandry techniques. Creating a captive environment that encourages natural behaviors may enhance survival in the wild following translocation.     

Rate of warming affects temperature sensitivity of anaerobic peat decomposition and greenhouse gas production

Temperature sensitivity of anaerobic carbon mineralization in wetlands remains poorly represented in most climate models and is especially unconstrained for warmer subtropical and tropical systems which account for a large proportion of global methane emissions. Several studies of experimental warming have documented thermal acclimation of soil respiration involving adjustments in microbial physiology or carbon use efficiency (CUE), with an initial decline in CUE with warming followed by a partial recovery in CUE at a later stage. The variable CUE implies that the rate of warming may impact microbial acclimation and the rate of carbon‐dioxide (CO₂) and methane (CH₄) production. Here, we assessed the effects of warming rate on the decomposition of subtropical peats, by applying either a large single‐step (10°C within a day) or a slow ramping (0.1°C/day for 100 days) temperature increase. The extent of thermal acclimation was tested by monitoring CO₂ and CH₄ production, CUE, and microbial biomass. Total gaseous C loss, CUE, and MBC were greater in the slow (ramp) warming treatment. However, greater values of CH₄–C:CO₂–C ratios lead to a greater global warming potential in the fast (step) warming treatment. The effect of gradual warming on decomposition was more pronounced in recalcitrant and nutrient‐limited soils. Stable carbon isotopes of CH₄ and CO₂ further indicated the possibility of different carbon processing pathways under the contrasting warming rates. Different responses in fast vs. slow warming treatment combined with different endpoints may indicate alternate pathways with long‐term consequences. Incorporations of experimental results into organic matter decomposition models suggest that parameter uncertainties in CUE and CH₄–C:CO₂–C ratios have a larger impact on long‐term soil organic carbon and global warming potential than uncertainty in model structure, and shows that particular rates of warming are central to understand the response of wetland soils to global climate change.     

Which species,how many,and from where: Integrating habitat suitability,population genomics,and abundance estimates into species reintroduction planning

Extirpated organisms are reintroduced into their former ranges worldwide to combat species declines and biodiversity losses. The growing field of reintroduction biology provides guiding principles for reestablishing populations, though criticisms remain regarding limited integration of initial planning, modeling frameworks, interdisciplinary collaborations, and multispecies approaches. We used an interdisciplinary, multispecies, quantitative framework to plan reintroductions of three fish species into Abrams Creek, Great Smoky Mountains National Park, USA. We first assessed the appropriateness of habitat at reintroduction sites for banded sculpin (Cottus carolinae), greenside darter (Etheostoma blennioides), and mottled sculpin (Cottus bairdii) using species distribution modeling. Next, we evaluated the relative suitability of nine potential source stock sites using population genomics, abundance estimates, and multiple‐criteria decision analysis (MCDA) based on known correlates of reintroduction success. Species distribution modeling identified mottled sculpin as a poor candidate, but banded sculpin and greenside darter as suitable candidates for reintroduction based on species‐habitat relationships and habitats available in Abrams Creek. Genotyping by sequencing revealed acceptable levels of genetic diversity at all candidate source stock sites, identified population clusters, and allowed for estimating the number of fish that should be included in translocations. Finally, MCDA highlighted priorities among candidate source stock sites that were most likely to yield successful reintroductions based on differential weightings of habitat assessment, population genomics, and the number of fish available for translocation. Our integrative approach represents a unification of multiple recent advancements in the field of reintroduction biology and highlights the benefit of shifting away from simply choosing nearby populations for translocation to an information‐based science with strong a priori planning coupled with several suggested posteriori monitoring objectives. Our framework can be applied to optimize reintroduction successes for a multitude of organisms and advances in the science of reintroduction biology by simultaneously addressing a variety of past criticisms of the field.     

Storage in the yolk platelets of low MW DNA produced by the regressing follicle cells.

The present work was carried out to clarify the nature and origin of the yolk DNA present in vitellogenic oocytes of the lizard Podarcis sicula. Morphological and biochemical evidences indicate that it has an intrafollicular origin, from the apoptotic bodies resulting from follicle cells regression at the end of previtellogenesis. This conclusion is reinforced by the observation that the oocyte membrane, in in vitro experiments, is unpermeable to exogenous DNA. Biochemical evidences reveal that the yolk DNA has a low (200bp) molecular weight and this suggests that it is produced by the endonucleases typically involved in apoptotic DNA laddering. Indeed, immunocytochemical analyses demonstrate that follicle cells contain significant amounts of DNAse I. In immunoblots, carried out during different periods of the ovarian cycle, the enzyme shows a MW of about 33, 66 or 100 kDa thus indicating that its activity in the follicle of Podarcis is modulated by dimerization and/or binding to regulatory factors. Mol. Reprod. Dev. 59: 422-430, 2001.     

DEVELOPMENT OF PROTOPLASTS OF ULVA FASCIATA (CHLOROPHYTA) FOR ALGAL SEED STOCK

The aim of this study was to isolate and cultivate protoplasts of the green alga Ulva fasciata Delile and subsequently induce them to form a microthallus suspension for algal seed stock. The protoplasts were covered with secreted mucilage following 6 h of culture when viewed with SEM. The mucilage fused to form thick layers during day 1 of culture. Microfibrillar cell walls were deposited into the thick layers of mucilage on the 5th day of culture. An average of about 10% of the freshly isolated protoplasts began to divide at 6–14 days. These protoplasts subsequently developed varied morphologies, depending on the time of collection during the year. Protoplasts isolated from U. fasciata collected in March to June developed frond thalli or microthalli when they were cultured in low or high densities (cells/area), respectively. The microthallus suspension was cultured for more than two years at 10–40 μ mol·m^{− 2} ·s^{− 1} . Frond thalli formed when the suspension was cultivated at 100–160 μ mol·m^{− 2} ·s^{− 1} . Therefore, microthallus suspension can serve as a seed stock of U. fasciata .     

Hunger and light level alter response to bait by Pacific halibut: laboratory analysis of detection, location and attack

Laboratory experiments with Pacific halibut Hippoglossus stenolepis revealed that hunger level had a significant effect on the first detection of bait, the number of baits located and attacked, the time required to locate and attack baits and handling times. In all cases, feeding motivation and efficiency increased with hunger. Light level influenced general locomotory activity and location and attacks on baits, but not detection or handling times. The effect of light was interactive with fish hunger level. Hungry fish could locate and consume baits in all light levels, ranging from daylight conditions to near darkness (10⁻⁸ µmoles photons m⁻² s⁻¹), but location, attack and handling times were all significantly elevated in low light conditions, and attack rates were significantly reduced. In the dark, only 50% of the baits were located and only 17% were attacked. Performance metrics were relatively similar among three higher light treatments (10⁻⁵, 10⁻³ and 10⁻¹ µmoles photons m⁻² s⁻¹) where bait location and attack were more efficient. Active space and effective area associated with baited fishing gear will vary because hunger and light levels affect variation in bait detection, locomotion and feeding behaviour. Consequently, fishing activity and stock assessments that depend upon bait may be compromised by spatial and temporal variation in prey abundance, time of day, season, depth and other environmental variables that influence feeding motivation and efficiency.     

Bird densities are associated with household densities

Increasing housing density is an important component of global land transformation, with major impacts on patterns of biodiversity. However, while there have been many studies of the changes in biodiversity across rural–urban gradients, which are influenced in large part by housing densities, how biodiversity changes across the full range of regional variation in housing density remains poorly understood. Here, we explore these relationships for the richness and abundance of breeding birds across Britain. Total richness, and that of 27 urban indicator species, increased from low to moderate household densities and then declined at greater household densities. The richness of all species increased initially faster with household density than did that of the urban indicator species, but nonurban indicator species richness declined consistently after peaking at a very low housing density. Avian abundance showed a rather different pattern. Total abundance and that summed across all urban indicator species increased over a wide range of household densities, and declined only at the highest household densities. The abundance of individual urban indicator species generally exhibited a hump-shaped relationship with housing density. While there was marked intraspecific variation in the form of such relationships, almost invariably avian abundance declined at housing densities below that at which the UK government requires new developments to be built. Our data highlight the difficulties of maintaining biodiversity while minimising land take for new development. High-density housing developments are associated with declines in many of those species otherwise best able to exploit urban environments, and those components of native biodiversity with which human populations are often most familiar.