Applicability of LIVE/DEAD BacLight stain with glutaraldehyde fixation for the measurement of bacterial abundance and viability in rainwater

Rainwater contains substantial bacteria and rain is an efficient pathway for the dissemination of bacteria from the atmosphere to land and water surfaces.However,quantitative information on rainwater bacteria is very limited due to the lack of a reliable method.In this study,the epifluorescence microscopy enumeration with the LIVE/DEAD BacLight Bacterial Viability Kit stain was verified to quantify the abundance of viable and non-viable bacterial cells in rainwater,with the 4',6-diamidino-2-phenylindole(DAPI) stain for the reference of total cell counts.Results showed that the total counts of bacterial cells by LIVE/DEAD BacLight staining were consistent with those by DAPI staining,and the average detection efficiency was(109 ± 29)%.The ratio of cell count with glutaraldehyde fixation to that without fixation was(106 ± 5)%on average.The bacterial concentration in negative control was usually an order of magnitude lower than that in rainwater samples.However,in case of small precipitation,the abundance in negative control could be more than that in rainwater samples.These results indicate that the enumeration with LIVE/DEAD BacLight bacterial viability assay coupled with glutaraldehyde fixation and careful negative control investigation is an approach applicable to the measurement of the concentration and viability of bacterial cells in rainwater.     

Review and appraisal of concept of sustainable food production systems

Environmental degradation, competition for resources, increasing food demands, and the integration of agriculture into the international economy threaten the sustainability of many food production systems. Despite these concerns, the concept of sustainable food production systems remains unclear, and recent attempts to appraise sustainability have been hampered by conceptual inconsistencies and the absence of workable definitions. Six perspectives are shown to underpin the concept. Environmental accounting identifies biophysical limits for agriculture. Sustained yield refers to output levels that can be maintained continuously. Carrying capacity defines maximum population levels that can be supported in perpetuity. Production unit viability refers to the capacity of primary producers to remain in agriculture. Product supply and security focuses on the adequacy of food supplies. Equity is concerned with the spatial and temporal distribution of products dervied from resource use. Many studies into sustainable agriculture cover more than one of these perspectives, indicating the concept is complex and embraces issues relating to the biophysical, social, and economic environments. Clarification of the concept would facilitate the development of frameworks and analytical systems for appraising the sustainability of food production systems. LRRC Contribution No. 90–46.     

Multiple song repertoire characteristics in the willow warbler (Phylloscopus trochilus): correlations with female choice and offspring viability

Male willow warblers have song repertoires which vary in complexity along several dimensions. We examined whether female choice, as measured by date of pairing, was based on these song characteristics in 4 different years. Pairing date was negatively correlated with song repertoire size in 1 year, and with song versatility in another year, but there was no consistent effect of any song characteristic on pairing throughout the years or in the pooled sample. The variable that best explained how soon a male pairs is male arrival date (only males that had settled territories before the first female arrived were considered in the analysis). This correlation is consistently significant in all years. This is most parsimoniously interpreted as females choosing some habitat characteristic in the same way that males do. A small percentage of males (8.3%) attracted and paired with a second female. The likelihood of becoming polygynous was not explained by any measured song characteristic, but it was related to arrival date: early males were more likely to pair with two females. Males with large repertoires fledged more young in their primary nests, and there was a trend for the offspring of these males to have a greater probability of being recruited into the population. In conclusion, the results show that in most years there is no sexual selection by female preference on song characteristics, although the data on reproductive success is consistent with the idea of repertoire size being an indicator of male quality. Received: 4 June 1999 / Received in revised form: 1 December 1999 / Accepted: 31 December 1999     

Using mRNA to investigate the effect of low-pressure ultraviolet disinfection on the viability of E. coli

UV can induce damages on mRNA consistently among different genes. SOS response was more active after UV treatment. Programmed cell death was not found to be more active after UV treatment. The efficacy of ultraviolet (UV) disinfection has been analyzed and validated by numerous studies using culture-based methods, yet the discovery of the viable but nonculturable state necessitates the investigation of UV disinfection based on viability parameters. Paired regulators of the SOS response system, recA-lexA, and the programmed cell death system, mazEF, in Escherichia coli were chosen as the target genes, and the effect of UV irradiation on the mRNAs of the four genes was studied. This research showed that, after UV irradiation, the responses of the mRNAs were highly consistent, with reduction percentages of approximately 60% at 20 mJ/cm², 70% at 40 mJ/cm², and 90% at 80 mJ/cm², and these reductions were believed to be the result of direct UV damage to nucleic acids. After 24 h of dark incubation, recA and lexA were both upregulated but to a lesser extent for repressor lexA; and mazE and mazF were both downregulated. This result implies that UV irradiation induces the dark repair system more actively, and the cells will proceed to death at a rate similar to that associated with natural decay.     

Interspecific mating of the introduced bumblebee <Emphasis Type="Italic">Bombus terrestris</Emphasis> and the native Japanese bumblebee <Emphasis Type="Italic">Bombus hypocrita sapporoensis</Emphasis> results in inviable hybrids

The bumblebee Bombus terrestris is not only an effective pollinator, but also a potential invasive alien species outside its native range. Recently, nearly 30% of queens of the Japanese native species Bombus hypocrita sapporoensis and B. hypocrita hypocrita were estimated to copulate with B. terrestris males in the field, suggesting that indigenous bumblebees could be genetically deteriorated through hybrid production with the introduced species. In this study, we evaluated hybrid production between the introduced B. terrestris and the indigenous B. hypocrita sapporoensis under laboratory conditions. The hatching rate of eggs derived from interspecific matings was 0% and 8.6% depending on the direction of the cross, which was significantly lower than that from intraspecific matings of B. terrestris (76.9%) and B. hypocrita sapporoensis (78.9%). Genetic studies using microsatellite markers revealed that both haploid and diploid individuals were present in the egg stage, whereas all hatched larvae were haploid. In addition, histological studies revealed that eggs derived from interspecific matings terminated development 2 days after oviposition. These results strongly suggested that eggs derived from interspecific matings are inviable due to post-mating isolation mechanisms. Mass release of exotic pollinators could cause serious population declines of native bumblebee species.     

Assessment of toxicity of a glyphosate-based formulation using bacterial systems in lake water

A new Aeromonas bioassay is described to assess the potential harmful effects of the glyphosate-based herbicide, Roundup^®, in the Albufera lake, a protected area near Valencia. Viability markers as membrane integrity, culturability and β-galactosidase production of Aeromonas caviae were studied to determine the influence of the herbicide in the bacterial cells. Data from the multifactor analysis of variance test showed no significant differences (P > 0.05) between A. caviae counts of viability markers at the studied concentrations (0, 50 and 100 mg l⁻¹ of glyphosate).

The effects of Roundup^® on microbial biota present in the lake were assessed by measuring the number of indigenous mesophilic Aeromonas in presence of different amounts of the herbicide at 0, 50 and 100 mg l⁻¹ of glyphosate. In samples containing 50 and 100 mg l⁻¹ of glyphosate a significant (P < 0.05) increase in Aeromonas spp. counts and accompanying flora was observed.

The acute toxicity of Roundup^® and of Roundup^® diluted with Albufera lake water to Microtox^® luminescent bacterium (Vibrio fischeri) also was determined. The EC₅₀ values obtained were 36.4 mg l⁻¹ and 64.0 mg l⁻¹ of glyphosate respectively. The acidity (pH 4.5) of the herbicide formulation was the responsible of the observed toxicity.     

Revisiting Evidence for Sustainability of Bushmeat Hunting in West Africa

Bushmeat hunting, a key source of dietary protein, has been implicated as a major extinction threat to tropical vertebrate species in West Africa. Ideally, any such hunting of wild species should be done sustainably, with off-take levels low enough to ensure viability of harvested species. Recent work purports to show that a mature bushmeat market in a major city in Ghana operates sustainably after depletion of vulnerable, slow-reproducing species (Cowlishaw and others 2005). I revisit two aspects of this work. First, I retest the prediction that larger species are transported to the market from greater distances, as expected if overexploitation depletes large species close to the city. Cowlishaw and others failed to find a significantly positive relationship between species-specific body mass and distance between capture site and the market. However, my reanalysis provides evidence for a positive relationship after all, consistent with unsustainable harvesting. In particular, ungulate species were harvested significantly farther from the market than smaller-bodied rodent species. Second, I caution that just because species “persist” in the marketplace in no way implies that they can withstand hunting pressure elsewhere and so should be of little concern to conservationists. I reveal that such species, despite their high intrinsic rates of population growth, are not robust elsewhere. Several of them have disappeared from a network of protected areas in Ghana (Brashares and others 2001). I show that faster-reproducing species are not necessarily more likely to persist in protected areas. The mere presence of fast-reproducing species in a mature bushmeat market should not be construed as generalizable robustness; criteria for ecological sustainability should ensure viability; and harvested species should be robust, not highly prone to extinction, in protected areas.     

Estimating the biogenic enhancement factor of weathering using an inverse viability method

To determine the influence of the biosphere on weathering we use a dynamic model of the global carbon cycle. It takes into account the most important processes for the long-term evolution of the Earth. The model is solved under a slowly changing environment of increasing solar luminosity and volcanic activity and continental area. By comparing the model results for the global average temperature with data derived from δ¹⁸O values from cherts it is possible to quantify the biogenic enhancement factor of weathering. For this purpose a newly developed inverse viability method is applied, which allows for calculating the range of possible biogenic enhancement factors consistent with the data. We find that in the Precambrian the weathering was 5.4 times lower than in the Phanerozoic era. This supports the hypothesis that the Cambrian explosion was caused by a positive feedback between the spread of biosphere, increased silicate weathering, and a consequent cooling of the climate.     

Sexual signalling and viability in a wolf spider (Hygrolycosa rubrofasciata): measurements under laboratory and field conditions

This study examined the crucial prediction of the conditional-handicap theory, the relationship between male sexual trait size and male viability, in the wolf spider Hygrolycosa rubrofasciata. In this species, males court females by drumming dry leaves with their abdomen, and males with the highest drumming rate enjoy highest mating success. We determined male drumming rate, body mass, and mobility, which reflects mate-searching activity, in relation to male survival. Because it is often difficult to know how results obtained from laboratory studies reflect the natural world, particularly when the measured variable is survival, we repeated our study in both laboratory and field conditions. Males drumming at the highest rate survived better than males drumming at a lower rate in both laboratory and field conditions. These results are in accordance with the predictions of conditional-handicap models of sexual selection. Survival was independent of male body mass and there was no significant correlation between male drumming activity and body mass. However, large males moved further than smaller males, and males moving longer distances lost less mass than those moving shorter distances. These results suggest that, moving, and consequently mate-searching activity, may be a condition-dependent trait and that there may be a advantage for large males in mate searching. Received: 22 October 1998 / Received in revised form: 28 January 1999 / Accepted: 14 February 1999     

Optimum Methods of Inactivation of Strongyloides stercoralis Larvae from Reclaimed Wastewater

Strongyloidiasis presents a major health hazard when reusing wastewater. Albendazol with a concentration of 4 mg/l, a contact time of 45 min, pH 1.2 and pH 10.2, killed the larva. The larva was also killed with a 0.03% concentration of iodine and a detention time of 30 min or a 0.04% concentration of iodine and a contact time of 10 min. The required detention time to inactivate larva in water was found to be 17 days. The minimum revealed residual chlorine to kill the larvae was 4 mg/l with 120 min of detention time, and a minimum contact time was found to be 80 min with a residual of 23 mg/l. Many methods are available to inactivate the Strongyloides stercoralis, which could be acids, alkaline chemicals, larvicidal chemicals, super chlorination, or just detention time. The best method must be determined according to the prevailing specific environmental site and the feasibility of the selected method(s).