Ornithine Decarboxylase in Trypanosoma brucei brucei: Evidence for Selective Toxicity of Difluoromethylornithine1

ABSTRACT. Activity of ornithine decarboxylase, the major rate limiting enzyme of polyamine biosynthesis, was determined in bloodstream trypomastigotes of Trypanosoma brucei brucei. The enzyme required pyridoxal-5′-phosphate, dithiothreitol and EDTA for optimal activity. Several properties of the enzyme were investigated and compared to the mammalian enzyme. Most notably, the parasite enzyme was >60-fold more sensitive to the inhibitor DL-α-difluoromethylornithine than its mammalian counterpart, thus making it an attractive target for chemotherapy.     

Synthesis and Content of Polyamines in Bloodstream Trypanosoma brucei*

SYNOPSIS. The sensitive dansyl procedure was used to detect putrescine and spermidine, but not spermine and cadaverine, in pleomorphic Trypanosoma brucei. The polyamines were synthesized in vitro from [³H]ornithine, [¹⁴C]arginine and [¹⁴C]methionine. Proline, agmatine, and citrulline, but not glutamine, glutamic or pyroglutamic acids, stimulated spermidine formation from [¹⁴C]methionine. Putrescine and spermidine synthesis occurred rapidly from ornithine: putrescine synthesis peaked in 0.5 h, spermidine in 1 h. Trypanosoma brucei assimilated exogenous ¹⁴C-labeled putrescine, spermidine, and spermine; spermidine and spermine were taken up 5 times as rapidly as putrescine. Polyamine syntheses may therefore be a practical target for novel trypanocies.     

PATTERNS OF ORIENTATION IN UNIONIDS AS A FUNCTION OF RIVERS WITH DIFFERING HYDROLOGICAL VARIABILITY

As unionids can become dislodged with high flows, it may provebeneficial for an individual to minimize its exposure to theflow. This can be accomplished by either burrowing as deep aspossible or orienting in a way that effectively reduces thedrag exerted on the mussel by the flow. The patterns of orientationwere examined in unionids with respect to hydrological variability.The orientation of mussels to flow was measured at four sitesalong an event river (hydro-logically variable) and a stableriver (hydrologically stable). Burrowing depth was measuredat a reference site in each river. Most individuals in bothriver types were oriented with their siphons pointing upstream.Mussel orientation differed significantly between the two rivertypes with mussels in the event river orienting more parallelto the flow than those in the stable river. Mussels in the eventriver were significantly larger than those in the stable riverbut the size of a mussel did not determine its orientation withina river. Burrowing depth did not differ for mussels betweenevent and stable rivers. The observed differences in orientationsamong river types are likely a function of differences in thepattern of orientation of the mussel community as a whole, withineach drainage. This burrowing behaviour may be an attributethat enhances the adaptations mussels have for remaining burrowedin the sediment. (Received 2 December 1996; accepted 24 February 1997)     

Associations between anuran tadpoles and salinity in a landscape mosaic of wetlands impacted by secondary salinisation

1. The secondary salinisation of wetlands is a global problem that poses a great threat to most freshwater biodiversity, including amphibians. We examined tadpole diversity in relation to wetland conductivity (our proxy for salinity) in wetlands in south‐eastern Australia to better understand (i) how salinity and amphibian diversity interact and (ii) the threat posed by secondary salinisation. 2. Six tadpole species were trapped in 56 wetlands that reflected a typical salinity gradient for the study region. We developed Bayesian models to examine the relationships between conductivity and both the probability of species occupancy and expected number of species with the imperfect detection probability of species accounted for in the models. 3. The probability of occupancy for all species and expected species number was negatively associated with wetland conductivity. Our results predict that conductivity should not limit tadpole presence below about 3000 μS cm⁻¹ at 25 °C (approximately 6% seawater) in the region, but will largely exclude amphibian larvae beyond about 6000 μS cm⁻¹ at 25 °C (approximately 12% seawater). 4. We also detected subtle among‐species differences in salinity tolerance. The results reported here show that tadpoles in the study region are likely to be negatively affected by projected future increases in salinisation.     

Acid Phosphatase in the Digestive Vacuoles and Lysosomes of Paramecium caudatum: A Timed Study1

Enzyme cytochemistry was used to determine when acid phosphatase (AcPase) becomes associated with the digestive vacuoles (DVs) of axenically grown Paramecium caudatum that were pulsed with latex beads for 2–3 min. When cells were incubated in the Gomori medium, AcPase was not observed in the discoidal vesicles, the acidosomes, and the newly released DVs up to 3 min old or in most DVs 3–6 min old. The number of AcPase-positive DVs increased to 56% when DVs were 12–18 min old. Similar results were obtained using the napthol AS-TR phosphate-hexaotized rosanilin method at the light microscopic level where hundreds of DVs were scored though the maximal level of positive DVs obtained by this method was lower. In addition to DVs of specific ages, AcPase was found in ER, in some Golgi vesicles, and small vesicles similar in diameter to Golgi vesicles which may represent primary lysosomes in this ciliate. Larger vesicles abundant near the DV-II were only partially filled with reaction product. These vesicles, which could be identified by their paracrystalline sheets and a prominent glycocalyx lining the luminal surface of their membranes, fit the definition for secondary lysosomes. These results, which indicate that lysosomes fuse with DVs only after they have attained a certain age, suggest the existence of specific recognition factors on the membranes of secondary lysosomes as well as DV-II.     

APPLIED ISSUES: Exploring the response of functional indicators of stream health to land‐use gradients

1. Broad‐scale assessment of stream health is often based on correlative relationships between catchment land‐use categories and measurements of stream biota or water chemistry. Few studies have attempted to characterise the response curves that describe how measures of ecosystem function change along gradients of catchment land use, or explored how these responses vary at broad spatial scales. 2. In autumn 2008, we conducted a survey of 84 streams in three bioregions of New Zealand to assess the sensitivity of functional indicators to three land‐use gradients: percentage of native vegetation cover, percentage of impervious cover (IC) and predicted nitrogen (N) concentration. We examined these relationships using general linear models and boosted regression trees to explore monotonic, non‐monotonic and potential threshold components of the response curves. 3. When viewing the responses to individual land‐use gradients, four of five functional indicators were positively correlated with the removal of native vegetation cover and N. In general, weaker and less responsive models were observed for the IC gradient. An analysis of the response to multiple stressors showed δ¹⁵N of primary consumers and gross primary productivity (GPP) to be the most responsive functional indicators to land‐use gradients. The multivariate models identified thresholds for change in the relationship between the functional indicators and all three land‐use gradients. Apparent thresholds were <10%IC, between 40 and 80% loss of native vegetation cover and at 0.5 and 3.2 mg L^?1 N. 4. The strength of regression models and the nature of the response curves suggest that measures of ecosystem function exhibit predictable relationships with land use. Furthermore, the responses of functional indicators varied little among three bioregions. This information provides a strong argument for the inclusion of functional indicators in a holistic assessment of stream health.     

Attempting to Manage Complex Predator-Prey Interactions Fails to Avert Imminent Extinction of a Threatened New Zealand Skink Population

Abstract: The current primary threats to biodiversity on a global scale are species invasions and habitat modification. Management of vulnerable populations often involves a lengthy sequence of 1) research to identify threats and recommend management strategies, 2) active management, and 3) results monitoring to assess effectiveness of management. The last mainland population of the large, endemic New Zealand skink (Whitaker's skink [Cyclodina whitakeri]) provides an opportunity to test this process in a system where the synergistic effects between invasive species (introduced rodents and grasses) have predicted outcomes. A low abundance of Whitaker's skink at the Pukerua Bay Scientific Reserve in the 1980s prompted management recommendations to remove grazing stock and revegetate the site to simultaneously restore habitat and provide protection against introduced mammalian predators. Since Whitaker's skink have low detectability, it was recommended that sympatric copper skink (C. aenea) be used an indicator species of management effectiveness. Grazing stock were removed in 1987, but efforts to revegetate the site were ineffective. Long-term monitoring (1984-2006) of the Whitaker's skink population and 4 other sympatric lizard species within a 336-m² area at the site resulted in 1,693 lizard captures over 7,597 trap days. Whitaker's skink represented 2.8% of all captures in 1984-1988, but declined relative to other species to represent only 0.2% of captures in 2000-2006 (representing 2 individuals). Congeneric copper skink showed a similar decline with capture rates also approaching zero by 2006. Removing grazing stock did not result in an increased abundance of Whitaker's skink or copper skink through improved habitat quality, as was intended by the management recommendation. Instead, reduced grazing has allowed introduced seeding grasses to proliferate, which may have led to periodic rodent irruptions, supporting a guild of introduced mammalian predators and depleting populations of Whitaker's skink and copper skink. In this instance, attempted protection may have driven a vulnerable population towards extinction. We recommend investigating the feasibility of constructing a mammal-proof fence around the core Whitaker's skink habitat, as the last remaining management option to salvage the population.     

The Diplomonad Fish Parasite Spironucleus vortens Produces Hydrogen

ABSTRACT. The diplomonad fish parasite Spironucleus vortens causes major problems in aquaculture of ornamental fish, resulting in severe economic losses in the fish farming industry. The strain of S. vortens studied here was isolated from an angelfish and grown in Keister's modified TY‐I‐S33 medium. A membrane‐inlet mass spectrometer was employed to monitor, in a closed system, O₂, CO₂, and H₂. When introduced into air‐saturated buffer, S. vortens rapidly consumed O₂ at the average rate of 62±4 nmol/min/10⁷ cells and CO₂ was produced at 75±11 nmol/min/10⁷ cells. Hydrogen production began under microaerophilic conditions ([O₂]=33.±15 μM) at a rate of 77±7 nmol/min/10⁷ cells. Hydrogen production was inhibited by 62% immediately after adding 150 μM KCN to the reaction vessel, and by 50% at 0.24 μM CO, suggesting that an Fe‐only hydrogenase is responsible for H₂ production. Metronidazole (1 mM) inhibited H₂ production by 50%, while CO₂ production was not affected. This suggests that metronidazole may be reduced by an enzyme of the H₂ pathway, thus competing for electrons with H⁺.     

First record of an apparently rare fig wasp feeding strategy: obligate seed predation

1. Fig trees require host‐specific agaonid fig wasps for pollination, but their figs also support numerous non‐pollinating fig wasps (NPFW) that gall fig tissues or develop as parasitoids. 2. Ficus microcarpa L. is widely naturalised outside its native range and the most invasive fig tree species. Seed predators are widely used for the biological control of invasive plants, but no obligate seed predatory (as opposed to ovule or fig wall galling) NPFW have been recorded previously from any fig trees. 3. Philotrypesis NPFW are usually regarded as parasitoids or ‘inquilines’ (parasitoids that also eat plant material) of pollinator fig wasps, but the present study provides evidence that Philotrypesis taiwanensis, a NPFW associated with F. microcarpa, is an obligate seed predator: (i) adults emerge from seeds of typical appearance, with a surrounding elaiosome; (ii) it shows no preference for figs occupied by fig wasp species, other than the pollinator; (iii) it only develops in figs that contain pollinated ovules, avoiding figs occupied by an agaonid that fails to pollinate; (iv) larvae are distributed in layers where seeds are concentrated and (v) it has a negative impact on seed but not pollinator offspring numbers. 4. Philotrypesis is a large genus, and further species are likely to be seed predators.