Detection of six serotypes of botulinum neurotoxin using fluorogenic reporters

Methods that do not require animal sacrifice to detect botulinum neurotoxins (BoNTs) are critical for BoNT antagonist discovery and the advancement of quantitative assays for biodefense and pharmaceutical applications. Here we describe the development and optimization of fluorogenic reporters that detect the proteolytic activity of BoNT/A, B, D, E, F, and G serotypes in real time with femtomolar to picomolar sensitivity. Notably, the reporters can detect femtomolar concentrations of BoNT/A in 4 h and BoNT/E in 20 h, sensitivity that equals that of animal-based methods. The reporters can be used to determine the specific activity of BoNT preparations with intra- and inter-assay coefficients of variation of approximately 10%. Finally, we find that the greater sensitivity of our reporters compared with those used in other commercially available assays makes the former attractive candidates for high-throughput screening of BoNT antagonists.     

Singing in the face of death: male banded wrens Thryophilus pleurostictus sing more to playback in their last breeding season

Investment in signalling is subject to multiple trade-offs that vary with life-stage, leading to a complex relationship between survival and trait expression. We show a negative relationship between survival and song rate in response to simulated territorial intrusion in male banded wrens Thryophilus pleurostictus , and test several explanations for this association. (1) Male age failed to explain the association: though age affected song rate in a cross-sectional analysis, longitudinal analysis showed that individuals did not increase their song rate as they got older. Reconciling these results suggests differential selection against young males that respond to intrusion with low song rates. (2) Mortality costs of high song rates did not appear to explain the negative relationship between song rate and survival because, though song rate in response to playback was condition-dependent, high song rates in a different context did not appear to impose mortality costs. (3) High levels of territorial pressure may have increased mortality, but were not associated with high song rates in response to playback. (4) Since song rates did not increase with age, but tended to increase only in the last year of life, we tentatively suggest that the negative relationship between song rate and survival could represent a terminal investment in territorial defence by males in their final breeding season, though further work is needed to confirm this conclusion.     

Mechanisms of prey selection by predaceous stoneflies: roles of prey morphology,behavior and predator hunger

Summary Laboratory feeding experiments using Hesperoperla pacifica (Banks), Perlidae, and Megarcys signata (Hagen), Perlodidae, as predators and Baetis tricaudatus Dodds and Ephemerella altana Allen as prey indicate a strong effect of prey morphology and mobility and predator hunger on prey selection by stoneflies. Knowledge of both dietary composition and feeding behavior was necessary to fully understand prey selection by these stoneflies.Fasted stoneflies presented with live prey ate more E. altana while satiated stoneflies ate approximately equal numbers of the two mayfly species. This pattern of dietary composition was the result of a reduction of attack frequency on the slower swimming E. altana with predator satiation and a continued high attack rate on B. tricaudatus regardless of recent feeding history. In contrast, fasted H. pacifica fed fresh frozen mayflies ate more B. tricaudatus indicating the importance of differences in prey mobility in controlling dietary composition.The high degree of similarity in patterns of feeding and mechanisms underlying those patterns for H. pacifica and M. signata suggest that they may be using similar rules for choosing mayfly prey and we suggest that mayfly prey are ranked by stoneflies on the basis of handling times. A general mechanistic model for stoneflies feeding on mayflies is presented and discussed.     

Trichoderma spp. and Bacillus spp. as growth promoters in maize (Zea mays L.)

Microbes that are beneficial to plants are used to enhance the crop growth, yield and are alternatives to chemical fertilizers. Trichoderma and Bacillus are the predominant plant growth-promoting fungi and bacteria. The objective of this study was select, characterize, and evaluate isolates of Trichoderma spp. and Bacillus spp. native from the northern region of Sinaloa, Mexico, and assess their effect on growth promotion in maize (Zea mays L.). In greenhouse conditions, four Trichoderma isolates and twenty Bacillus isolates, as well as two controls, were tested in a completely randomized design with three replicates. We selected the two best strains of Trichoderma and Bacillus: TB = Trichoderma asperellum, TF = Trichoderma virens, B14 = Bacillus cereus sensu lato and B17 = Bacillus cereus, which were evaluated in the field in a completely randomized blocks in factorial arrangement design with three replicates applying different rates of nitrogen fertilizer (0, 150 kg N/ha, and 300 kg N/ha). Treatments 5 (B17 = B. cereus) and 11 (TF = T. virens) both fertilized with 150 kg N/ha showed similar yields and they did not reveal significant differences from the treatments fertilized with 300 kg N/ha. This indicated that treatment 5 (B17= B. cereus with 150 kg N/ha) and treatment 11 (TF= T. virens with 150 kg N/ha) were efficient as growth promoters, by not showing significant differences in root volume and dry weight of foliage. The results indicated a reduction of 50% in the rate of nitrogen to fertilizer required for maize (Zea mays L.) crops. These microorganisms Trichoderma and Bacillus could be an alternative to reduce the use of chemical fertilizers in maize.     

Spatial and temporal variation of fish communities in secondary channels of the San Juan River, New Mexico and Utah

Spatial and temporal variation of fish communities in four secondary channels of the San Juan River between Shiprock, NM and Bluff, UT were investigated from July 1993 through November 1994. Fish abundance and habitat availability data were collected to determine if physical attributes of sites influenced spatial and temporal variation in their fish communities. Stability of habitat was shown to positively influence the stability of the fish community. Analysis of variance revealed greater spatial than temporal variation in the abundance of red shiner, Cyprinella lutrensis, fathead minnow, Pimephales promelas, and flannelmouth sucker Catostomus latipinnis, while speckled dace, Rhinichthys osculus showed greater temporal variation. Ordination, using detrended correspondence analysis, revealed variation in fish communities by site, date, and sample year. Spatial variation was greatest during low-flow periods when the greatest differences in habitat among the four sites occurred. Spring runoff had the greatest temporal effect on the fish communities in secondary channels and appeared to reset the communities by displacing those species that were less resistant to increased current velocities. This annual event may help maintain native fish species adapted to these conditions in the San Juan River while moderating the abundance of nonnative fish species.     

Influence of Experimental Flooding on Litter Dynamics in a Rio Grande Riparian Forest,New Mexico

Flow regulation, which has largely eliminated flooding along the Rio Grande in central New Mexico, has substantially changed the riparian ecosystem. We investigated managed flooding as a means of restoring ecosystem function. After collecting baseline data during 1991 and 1992 in two riparian forest sites that had not flooded for about 50 years, we flooded an experimental site for 27–32 days during late spring of 1993, 1994, and 1995, leaving the reference site unflooded. During the final year of the study we compared these sites to two additional sites located within the riverside levee, one of which has been flooding naturally while the second remained largely unflooded. Three years of experimental flooding did not change the total biomass of either woody debris or forest-floor litter at the experimental flood site. Both woody debris and forest-floor litter, however, were significantly lower at the natural flood site than at the experimental flood site and two unflooded sites. Leaf and wood decomposition rates increased with flooding. The decay rate for cottonwood logs at the unflooded site (0.010 per year) predicted a half-life of 69.3 years, while the decay rate of 0.065 per year after 3 years of experimental flooding predicted a half-life of 10.6 years. This suggests that a decade of annual flooding may be used to restore the organic debris to pre-regulation levels. Flooding may also have caused an increase in litter production. These results suggest that experimental flooding has initiated a process of restoring ecosystem function within the riparian forest.     

Nile perch and the transformation of Lake Victoria

The transformation of Lake Victoria that began in 1980 followed the population explosion of Nile perch Lates niloticus, causing the apparent extirpation of 500+ endemic haplochromine species and dramatic physico-chemical changes. Officially introduced in 1962–1963, but present earlier, the reasons for the long delay before its population exploded are discussed. The hypothesis that it occurred only after the haplochromine decline is evaluated, but haplochromines declined only after the Nile perch expansion began. The sudden eutrophication of the lake was attributed to Nile perch, but evidence of eutrophication from 1950 onwards led some researchers to conclude that it was the result of climatic changes. We conclude that the haplochromine destruction disrupted the complex food webs that existed prior to the upsurge of Nile perch. The depletion of fish biomass by Nile perch may have been the source of extra phosphorus responsible for the eutrophication of the lake. After the Nile perch explosion in 1980 the fish population came to be dominated by only three species, but fisheries productivity increased at least 10-fold. Fishing has caused demographic changes in Nile perch, which may have allowed some haplochromine species to recover. The condition of the lake appears to have stabilised since 2000, partly because the fish biomass has risen to at least 2 × 10⁶ t, replacing the ‘lost’ biomass and restoring some ecosystem functioning.     

Biodiversity monitoring by community‐based restoration groups in New Zealand

In recent decades, community groups have transformed habitat restoration, pest control and species translocations in New Zealand. Large areas of wild New Zealand benefit hugely from ongoing management by community‐based restoration groups. Areas near cities and towns have especially good access to pools of keen volunteers. Community groups are involved in monitoring progress with their work, as well as monitoring biodiversity changes in general at their project sites. New tools powered by modern technologies are creating the opportunity for New Zealand's community volunteers to play a transformative role in biodiversity monitoring for either purpose. These tools are reducing the resources and expertise required for species detection and identification. Smartphones with cameras, GPS, audio recorders and data apps make it easier than ever to record species observations. Crowd‐sourced identification of species in photographs and sounds loaded onto NatureWatch NZ allow volunteers to make observations of a much wider range of taxa than just common birds and trees. Realising this potential requires community groups, scientists and their institutions to collaborate in building and maintaining simple, accessible monitoring systems that (i) require and promote standard monitoring methods, (ii) provide efficient data entry in standard formats, (iii) generate automated results of use to community groups and (iv) facilitate public sharing of data to contribute to regional, national and global biodiversity monitoring. Some New Zealand monitoring systems developed recently to assist community‐based restoration groups with monitoring mammalian predator control are good examples of this approach. Making this happen at a large scale across many community groups and taxa requires increased and coordinated long‐term institutional support for monitoring systems and training.