Harnessing the potential for otolith microchemistry to foster the conservation of Amazonian fishes

1. Freshwater environments host roughly half of the world’s fish diversity, much of which is concentrated in large, tropical river systems such as the Amazon. Fishes are critical to ecosystem functioning in the Amazon River basin but face increasing human threats. The basic biology of these species, and particularly migratory behaviour, remains poorly studied, in part owing to the difficulty associated with conducting tagging studies in remote tropical regions.
2. Otolith microchemistry can circumvent logistical issues and is an increasingly important tool for studying fish life histories. However, this approach is still new in the Amazon, and its potential and limitations to inform fish conservation strategies remain unclear.
3. Here, otolith microchemistry studies in the Amazon are reviewed, highlighting current possibilities, and several key factors that limit its use as a conservation tool in the Amazon are discussed. These include the dearth of spatiotemporal elemental data, poor understanding of environment–fish–otolith pathways, and insufficient funding, facilities, and equipment.
4. A research initiative is proposed to harness the potential of this technique to support conservation in the Amazon. Key aspects of the proposal include recommendations for internal and external funding, which are critical to acquiring and maintaining technical staff, cutting-edge equipment, and facilities, as well as fostering regular scientific meetings and working groups. Meetings can facilitate a systematic approach to investigating environment–otolith pathways, broadening the chemical baseline for most Amazonian tributaries, and exploring potential valuable elements.
5. These outcomes are urgently needed to conserve biodiversity and ecosystem functioning in the Amazon, especially given threats such as widespread hydroelectric damming. The initiative proposed here could make otolith microchemistry an important, cost-effective tool to inform and foster conservation in the Amazon, and act as a template for other imperilled tropical river basins, such as the Mekong and the Congo.

     

Detection of Flexibacter maritimus in fish tissue using nested PCR amplification

A nested polymerase chain reaction (PCR) system was developed for the detection of Flexibacter maritimus from fish tissue. The total procedure for the diagnosis of marine flexibacteriosis, from the point of DNA extraction to the electrophoretic analysis, can be performed in < 4 h. This was achieved by the combination of a short thermal cycling programme with a rapid DNA extraction procedure. The assay was extremely sensitive, capable of detecting as few as 75 cfu mg(-1) fish tissue. The accuracy of the nested PCR was confirmed under field conditions using tissue samples recovered during 1993-2002 from fish suffering marine flexibacteriosis. The nested PCR method proved to be efficient for the rapid and sensitive detection of F. maritimus from fish tissues and can be used for routine diagnosis of the disease caused by this pathogen.     

Osmoregulation in juvenile Siberian sturgeon (Acipenser baerii)

The osmoregulation capabilities of juvenile Siberian sturgeon exposed to three experimental osmolalities (22, 250 and 387 mOsmol kg⁻¹) were studied over a 45-day period. Growth performance, haematological parameters, ion concentrations, gill and spiral valve Na⁺-K⁺-ATPase activities, as well as gill and spiral valve histology, were measured. At the end of the period, the plasma osmolality of fish kept in 250 and 387 mOsmol kg⁻¹ was higher than that of fish kept in 22 mOsmol kg⁻¹. Similar trends were observed in electrolyte concentrations. Spiral valve and gill Na⁺-K⁺-ATPase activity varied with exposure time and environmental salinity. Shortly after being transferred to hyperosmotic media, spiral valve Na⁺-K⁺-ATPase activity fell, while gill Na⁺-K⁺-ATPase activity remained constant. At the end of the experiment, gill Na⁺-K⁺-ATPase activities in fish kept in isosmotic and hyperosmotic media had increased in comparison to those of the control fish. Moreover, spiral valve Na⁺-K⁺-ATPase activities recovered and were similar to those recorded in fish kept in hyposmotic environments. Although some of the morpho-physiological mechanisms were operational in juvenile Siberian sturgeon in their adaptation to hyperosmotic media, fish cannot be considered hyperosmotic regulators as they were unable to maintain their plasma osmolality and electrolyte equilibrium in salinities higher than 250 mOsmol kg⁻¹. This suggests that the culture of juvenile Siberian sturgeon in brackish environments is unlikely to be successful. However, our data indicate that in natural environments, juvenile Siberian sturgeon in migratory populations (Ob and Lena Rivers) would be able to migrate successfully into estuarine brackish grounds with a salinity of up to 9%. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification and characterization of lactic acid bacteria isolated from rainbow trout, Oncorhynchus mykiss (Walbaum), with inhibitory activity against Lactococcus garvieae

A study was conducted to evaluate the probiotic properties of endogenous rainbow trout microbiota against pathogenic Lactococcus garvieae. A total of 335 bacterial strains were isolated from rainbow trout and screened for antagonistic activity against L. garvieae using an agar spot assay. Antagonistic strains were grouped by PCR amplification of repetitive bacterial DNA elements (rep‐PCR) and identified by 16S rRNA gene sequence analysis. The results revealed that the antagonistic strains belonged to the genera Lactobacillus, Lactococcus and Leuconostoc. Further probiotic characteristics, such as specific growth rate, doubling time, resistance to biological barriers, antibiotic resistance, hydrophobicity and production of antimicrobial substances, were also studied. These strains were able to survive low pH and high bile concentrations, showed good adherence characteristics and a broad spectrum of antibiotic resistance. The antagonistic efficacy was maintained after sterile filtration and was sensitive to proteinase K, indicating that proteinaceous extracellular inhibitory compounds were at least partially responsible for pathogen antagonism. Based on these results, these strains should be further studied to explore their probiotic effects in challenge experiments in vivo. This study shows clear evidence that the indigenous trout‐associated microbiota may provide a defensive barrier against L. garvieae.     

Bioavailability of organoclay formulations of atrazine in soil

Pesticide formulations based on organoclays have been proposed to prolong the efficacy and reduce the environmental impact of pesticides in soil. This research addressed the question of whether atrazine in organoclay-based formulations is irreversibly sorbed or is bioavailable for bacterial degradation in soil. Different cations of l-carnitine (CAR), tyramine (TYRAM), hexadimethrine (HEXADIM), phenyltrimethylammonium (PTMA), hexadecyltrimethylammonium (HDTMA), and Fe(III) were incorporated into Na-rich Wyoming montmorillonite (SWy-2) and Ca-rich Arizona montmorillonite (SAz-1) at 100% of the cation exchange capacity (CEC) of the clays as a strategy to enhance the affinity of the clay minerals for atrazine. A Buse loam soil from Becker, MN, was treated with three organoclay-based formulations of 14C-atrazine or free herbicide and incubated for 2 weeks. To determine the bioavailability of 14C-atrazine, the soil was inoculated with Pseudomonas sp. strain ADP, which rapidly mineralizes atrazine. At day 0, and after a 2 week incubation, mineralization and the amount of 14C-atrazine residues distributed between the aqueous-extractable, methanol-extractable, and bound fractions in the soil were determined to characterize the availability of nonaged and aged atrazine residues. By the end of the 2 week incubation, the microorganisms had mineralized >80% of the initial readily available (water-extractable) and >70% of the less readily available (methanol-extractable) 14C-atrazine in the soil. Bound residues increased from <4% at day 0 to ～17% after the 2 week incubation for both the formulated and free forms of atrazine. The results of these incubation experiments show that the bioavailabilities of atrazine were similar in the case of the organoclay formulations and as free atrazine. This indicated that whereas more atrazine was sorbed and less likely to be transported in soil, when formulated as organoclay complexes, it was ultimately accessible to degrading bacteria, so that the herbicide is likely to be naturally attenuated by soil microorganisms.     

Population trends in northern spotted owls: Associations with climate in the Pacific Northwest

We used reverse time capture-mark-recapture models to describe associations between rate of population change (λ) and climate for northern spotted owls (Strix occidentalis caurina) at six long-term study areas in Washington and Oregon, USA. Populations in three of six areas showed strong evidence of declining populations, while populations in two additional areas were likely declining as well. At four areas, λ was positively associated with wetter-than-normal conditions during the growing season, which likely affects prey availability. Lambda was also negatively associated with cold, wet winters and nesting seasons, and the number of hot summer days. The amount of annual variation in λ accounted for by climate varied across study areas (3-85%). Rate of population change was more sensitive to adult survival than to recruitment; however, there was considerable variation among years and across study areas for all demographic rates. While annual survival was more closely related to regional climate conditions, recruitment was often associated with local weather. In addition to climate, declines in recruitment at four of six areas were associated with increased presence of barred owls. Climate change models predict warmer, wetter winters and hotter, drier summers for the Pacific Northwest in the first half of the 21st century. Our results indicate that these conditions have the potential to negatively affect annual survival, recruitment, and consequently population growth rates for northern spotted owls.     

Separation of olive proteins combining a simple extraction method and a selective capillary electrophoresis (CE) approach: application to raw and table olive samples

A simple extraction method was developed to extract proteins from olive samples based on chloroform/methanol extraction followed by a protein precipitation with cold acetone. Then, a capillary electrophoresis (CE) method was carried out using an acid buffer (1 M formic acid at pH 2) to ensure a positive net charge for proteins and a neutral charge for potential interferents as polyphenols. The method developed was applied to raw and table olive samples. Interestingly, raw olive samples showed differences in protein profiles depending upon the botanical variety of olives and their geographical region. Protein profiles obtained for table olives also showed differences according to the sample treatment. Thus, a signal reduction in the electropherograms obtained for black olives was observed in comparison to those achieved for treated green olives. In this work, the use of protein profiles was demonstrated to be a powerful tool for studying variations among olive samples.     

Fraxinus-Glomus-Pisolithus symbiosis: Plant growth and soil aggregation effects

It has been established that arbuscular mycorrhizal (AM) fungi are involved in the conservation of soil structure. However, the effect of ectomycorrhizal (EM) fungi alone or in interaction with AM fungi in soil structure has been much less studied. This experiment evaluated EM and AM fungi effects on soil aggregation and plant growth. Ash plants (Fraxinus uhdei) were grown in pots, and were inoculated with Glomus intraradices and Pisolithus tinctorius separately but also in combination. Our results showed that F. uhdei established a symbiotic association with EM and AM fungi, and that these organisms, when interacting, showed synergistic and additive effects on plant growth compared to singly inoculated treatments. EM and AM fungi prompted changes in root morphology and increased water-stable aggregates. AM fungi affect mainly small-sized macroaggregates, while EM and EM-AM fungi interaction mainly affected aggregates bigger than 0.5 mm diameter. These results suggest that ectomyccorrhizal as well as arbuscular mycorrhizal fungi should be considered in restoration programs with Fraxinus plants.     

Heavy Metal and Nitrogen Monitoring Using Moss and Topsoil Samples in a Pyrenean Forest Catchment

This study was carried out in order to assess both the deposition of heavy metal and nitrogen in a mountain ecosystem with low levels of metal deposition and its possible interactions with factors such as lithology and topography. For this purpose, samples of Hypnum cupressiforme Hedw. and topsoils were collected in a forest catchment within Bertiz Natural Park, an International Cooperative Programme on Integrated Monitoring site. Trace metals levels in mosses can be considered low compared with values reported elsewhere in Europe, and the dust soil mineral particles seemed to be the main source of these values. Only Cd and Hg presented external inputs, probably with an anthropic origin, for mosses according to the enrichment factor values, whereas historical pollution-related deposition in soils was determined for Pb, Cu, and Ni, attending to their total/extractable ratio.     

Bioaccumulation of Copper by Zea mays: Impact on Root, Shoot and Leaf Growth

In the present study, the growth and the Cu²⁺accumulation by roots, shoots and leaves of Zea mays were examined using copper sulphate in the range of 10^?4 to 10^?2 M. Plants of Z. mays did not show inhibition of growth in the presence of 10^?4 to 10^?2 M Cu²⁺; however, it was observed growth effects on root when different Cu²⁺ solution concentrations were used. Only the seedlings exposed to 10^?2 M exhibited substantial root growth reduction, yielding only 56% of length with respect to the control. Seedlings exposed to 10^?4 M Cu²⁺ exhibited 16% and 42% growth increase in shoots and leaves, respectively, when compared with the controls. The seedlings treated with 10^?3 and 10^?2 M Cu²⁺ were inhibited in shoot and leaf growth. The fresh weights in roots, shoots and leaves significantly decreased at 10^?2 M Cu²⁺. The tolerance index, based on root length, was not significantly different for the three different treatments with copper. However, the total accumulation rate was very low at 10^?4 and 10^?3 M compared to 10^?2 Cu treatments. The capacity of copper accumulation by roots, shoots and leaves of Z. mays plants increased concomitant to the copper concentration, arriving to 382 times more in roots, 157 in shoots and only 16 in leaves, all compared to the controls. Cu could be accumulated by roots, shoots and leaves when the initial concentrations were 10^?3 and 10^?4 M. However, when it was 10^?2 M, the metal could not be accumulated by leaf and shoot levels; the roots could increase their copper accumulation capacity three times compared to the control. Z. mays has potential ability to accumulate Cu without being overly sensitive to Cu toxicity.