Potential for biofilms as biological indicators in Australian riverine systems

Summary Biological indicators have been widely used in Australian riverine systems to assess the effectiveness of past and current management. The short generation time, sessile nature, responsiveness to environmental conditions and the availability of sound, quantitative methodologies make biofilms suitable as a monitoring tool in these systems. This paper describes biofilm structure, function and development through the processes of succession and disturbance. Biofilms are assemblages of algae, fungi and microorganisms which cover rocks, wood and sediments in aquatic systems. A review of biofilm collection and processing techniques using relevant Australian and international studies reveals a large literature on many structural and functional biofilm attributes. Studies using structural attributes such as biomass and diversity to examine water quality impacts and invertebrate grazers dominate the Australian literature. More recently, studies have used functional biofilm attributes such as metabolism and foodweb interactions. Monitoring programs that combine structural and functional biofilm attributes will allow the best assessment of impacts in riverine systems. Biofilm functional parameters provide an integrated, long-term measure of ecosystem function, with structural attributes such as biomass and diversity allowing historical comparisons with previously recorded datasets. Monitoring programs such as these with a well-founded scientific base and defined management outcomes will expand our knowledge of river function and contribute to the restoration of Australian river systems.     

Use of atrazine sensitive leguminous plants as biological indicators to evaluate the atrazine degradation efficiency of a bacterial inoculum

Atrazine sensitive leguminous plants were grown in a soil spiked with atrazine and augmented with an atrazine-degrading bacterium, Arthrobacter sp. strain MCM B-436, to ascertain its degradative efficiency. Germination and survival of plants was correlated with atrazine removal from soil. This experiment was carried out at laboratory as well as field level, showing consistent results. This bioindicator approach serves as an efficient measure for atrazine removal and could be easily adapted to determine atrazine degradation efficiency of other microbial strains.     

Cilioprotists as biological indicators for estimating the efficiency of using Gravel Bed Hydroponics System in domestic wastewater treatment

Interest has increased over the last several years in using different methods for treating sewage. The rapid population growth in developing countries (Egypt, for example, with a population of more than 87 millions) has created significant sewage disposal problems. There is therefore a growing need for sewage treatment solutions with low energy requirements and using indigenous materials and skills. Gravel Bed Hydroponics (GBH) as a constructed wetland system for sewage treatment has been proved effective for sewage treatment in several Egyptian villages. The system provided an excellent environment for a wide range of species of ciliates (23 species) and these organisms were potentially very useful as biological indicators for various saprobic conditions. Moreover, the ciliates provided excellent means for estimating the efficiency of the system for sewage purification. Results affirmed the ability of this system to produce high quality effluent with sufficient microbial reduction to enable the production of irrigation quality water.     

Use of an ungulate‐specific feed structure as a potential tool for controlling feral goats in Australian forest ecosystems

The feral Goat (Capra hircus) has successfully exploited a range of landscapes around the world with occurrences of overabundance resulting in significant damage to ecological values. In forested ecosystems in Australia, there are currently limited means to control the species when compared to the range of management techniques available for other pest animals. To redress this deficiency, we designed a feed structure combined with commercially available salt blocks to attract goats to set locations in a forested study area. Structures that exploited differences in the pedal morphology (foot size and shape) of native herbivores (kangaroos and wallabies) and ungulates (feral goats and deer) were found to be highly target‐specific, with feral goats freely able to access salt blocks, whilst nontarget native species were effectively excluded. Other introduced ungulate species, Fallow Deer (Dama dama) and Red Deer (Cervus elaphus), successfully accessed salt blocks in feed structures but at a considerably lower rate than feral goats. The capacity to present a range of bait types within a target‐specific feed structure, once matched with a humane toxicant, could provide land managers with an additional cost‐effective lethal control tool for future management of feral ungulates, particularly goats.     

Specific ratio and survival of Pseudomonas CF600 as co-culture for phenol degradation in continuous cultivation

Studies were carried out to understand parallel survival of two strains when cultivated as co-culture on a single carbon source in continuous cultivation. Strains used were Pseudomonas sp. strain CF600 that is reported for degradation of phenol; and HKR1 a lab strain, which was isolated from a site contaminated with phenol. In continuous cultivation Pseudomonas sp. CF600 showed an accumulation of colored intermediate, 2-hydroxy muconic semialdehyde (HMS), when fed with phenol as a sole source of carbon under dissolved oxygen limiting condition (40% saturation level). Under the same cultivation condition when it was co-cultured with strain HKR1, complete degradation of phenol was observed with no accumulation of intermediate. Different dilution rates (0.03, 0.15, and 0.30) were set in the bioreactor during cultivation. It was also observed that both the strains follow a typical cell density ratio of 1:18 as strain HKR1: Pseudomonas sp. CF600 irrespective of the dilution rates used in the study to favor degradation of phenol. Pseudomonas sp. CF600 is reported to degrade phenol via a plasmid-encoded pathway (pVI150). The enzymes for this meta-cleavage pathway are clustered on 15 genes encoded by a single operon, the dmp operon. PCR using primers from the different catabolic loci of dmp operon, demonstrated that the strain HKR1 follows a different metabolic pathway for intermediate utilization.     

Performance evaluation of eight years experience of constructed wetland systems in Catalonia as alternative treatment for small communities

In Catalonia (Spain), a variety of different systems have been built to naturally treat liquid residues from small communities. Some of these wastewater treatment plants (WWTPs) include constructed wetlands with horizontal subsurface flow (HSSF) as secondary treatment. The present study described and characterized the performance of 11 WWTPs with secondary HSSF constructed wetland systems after an initial operating period of 8 years. The effluent concentrations of Biochemical Oxygen Demand (BOD₅), Total Suspended Solids (TSS), Total Nitrogen (TN) and Total Phosphorous (TP) were statistically analyzed, and removal efficiencies for all WWTPs including all stages in treatment were calculated. The accumulated probability functions of those parameters were evaluated to determine the influence of two different types of polishing units on the overall performance: (a) only lagoon systems and (b) lagoon systems with HSSF. The statistical analysis indicates good performance for BOD₅ and TSS. In the first case, mean concentrations below 25 mg/L were found in 9 of the 11 plants analyzed and removal efficiencies between 78 and 96% were observed. In the second case, mean concentrations below 35 mg/L were found in 8 of the 11 plants, and removal efficiencies were between 65 and 88%. For the nutrients, the removal efficiency for TN and TP were in the range of 48-66% and 39-58%, respectively. Additionally, the analysis of the influence of the polishing units did not show a significant improvement (α > 0.05) for any parameter in the wetland systems without a subsequent polishing unit. However, in the wetland systems with a polishing unit of HSSF, a significant improvement (α < 0.05) was found for the effluent's BOD₅, TN and TP concentrations but with no significant contribution in TSS management.     

Selective electrode for dibenzyl dimethyl amoonium cation as indicator of the membrane potential in biological systems

The electrode sensitive to dibenzyl dimethyl ammonium (DDA⁺), which is considered to be an indicator of the membrane potential, was constructed by using tetraphenyl borone (TPB^?) embedded in dichloroethane. Rapid and Nernstian responses were exhibited against DDA⁺ solutions ranging between 10^?2 and 3 · 10^?6 M in concentration. High selectivity for DDA⁺ was observed in the presence of various inorganic salts, ADP, ATP, oxidizable substrates and sugars. The electrode developed here was used to measure the DDA⁺ uptake in Streptococcus faecalis and the results agreed with those reported by Harold, F.M. and Papineau, D. ((1972) J. Membrane Biol. 8, 27–44 and 45–62). While they determined the DDA⁺ concentration in the medium by measuring the absorbance of the filtrate treated with the ion-exchangers, the electrode can measure directly the DDA⁺ concentration in the bacterial suspension without any pretreatment. It was also shown that the electrode can measure the DDA⁺ uptake in mitochondria during energization.     

An advanced Index of Biotic Integrity for use in tropical shallow lowland streams in Costa Rica: Fish assemblages as indicators of stream ecosystem health

This paper presents an advanced version of the Index of Biotic Integrity (IBI), a multimetric index to indicate ecosystem health. The multimetric index has been adapted in such a way that it not only indicates overall condition but also specific causes of environmental disturbance. The newly developed index (a) uses data of tolerant as well as intolerant species in a single metric to indicate environmental disturbance, (b) does not require knowledge about species from the literature, and (c) can be applied to artificial landscapes.The metrics proposed here consist of indicator species assemblages that are selected directly for their relationship with an environmental component or specific type of environmental degradation. Thus, each metric indicates a type of environmental concern, which enables conservation practices to be targeted more effectively. Species assemblages for each single metric consist of a combination of species that can be negatively and positively related to environmental disturbances, providing a better indication of stream ecosystem health.The area studied was assumed to be too diverse for one single index. Canonical Indirect-Gradient Principal Component Analysis indicated that the optimal division of subindices based on stream typology was for streams with drainage basin sizes <10 km² and >10 km². Pearson Product-Moment Correlations were used to identify relationships between anthropogenic disturbances and the composition and abundance of fish species at impacted as well as undisturbed sites. This index proved to be useful for indicating overall stream ecosystem health as well as local onsite environmental disturbances or the environmental components of greatest concern. This index does require extensive information about measured levels of anthropogenic disturbances with the accompanying composition and abundance of fish species.     

An immunoaffinity liquid chromatography-tandem mass spectrometry assay for detection of endogenous aggrecan fragments in biological fluids: Use as a biomarker for aggrecanase activity and cartilage degradation

The degradation of articular cartilage by aggrecanases (ADAMTS-4 and ADAMTS-5) plays a significant role in the pathology of osteoarthritis (OA). To monitor aggrecanase activity in OA, we have developed a sensitive, accurate, and versatile assay for detection of two specific cleavage sites on aggrecan. The assay uses an immunoaffinity-based liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to detect cleavage at the ³⁷⁴ARGS site and the ¹⁸²⁰AGEG site. The dynamic range of the assay is more than three orders of magnitude, with interassay precision less than 15%. It has been successfully applied to various biological fluids and species, including rat, bovine, dog, and human. The assay has been analytically qualified for use in human urine and synovial fluid (SF). The limits of detection (LODs) for ARGS in urine and SF are 2.5 and 10 pg/ml, respectively, whereas the LOD for AGEG is 20 pg/ml in SF. Analysis of these biomarkers from OA subjects and normal healthy volunteers revealed a significant elevation of both markers in OA. Similarly, in a rat model of cartilage degradation, both ARGS and AGEG were elevated, demonstrating the utility of these biomarkers for translational research. These data suggest that the ARGS and AGEG biomarkers developed have potential as measures of aggrecanase activity in OA and may contribute to our understanding of OA pathology.