Features and conservation of the Brazilian Pantanal wetland

The Pantanal is a 140,000 km² sedimentary floodplain in western Brazil and one of the largest wetlands in the world. The main landscapes and phytophysiognomies, according to flood origin, are briefly described and some of the characteristic plant species are mentioned: (a) river flood (1–5 m) on clayey eutrophic soils with gallery forests, pioneer forests and scrub, Tabebuia and Copernicia parks, seasonal swamps, grasslands and oxbow lakes; and (b) rain flood (10–80 cm) mainly on dystrophic sandy soils (72% of the total area) with savanna (cerrado) grasslands and woodlands, with or without ponds. Regulating factors of the vegetation such as wet-and-dry cycle and management are considered. Dynamics of the vegetation, in particular the aquatic types, are shortly depicted. The role of grazing for conservation is discussed, and we suggest that 200 years of cattle ranching apparently did not cause major changes in the vegetation, except turning tall grass into short swards, as the domestic herd found a nearly empty niche. However, severe threats to the flora and fauna of the Pantanal originate outside the floodplain. Siltation of the Taquari river is pointed out as the worst environmental problem, changing the hydrology (wet-and-dry to wet), fauna and flora, e.g. eliminating riparian forest.This revised version was published online in March 2005 with corrections to the issue cover date.     

Photosynthetic parameters of two invasive tree species of the Brazilian Pantanal in response to seasonal flooding

The Pantanal is the largest wetland in the world with extremely high plant and animal diversity, but large areas have been invaded by Vochysia divergens Pohl (Vochysiaceae), a tree that is native to the Amazon Basin, and Curatella americana L. (Dilleniaceae), a tree that is native to the Brazilian savanna (cerrado). V. divergens is reportedly floodadapted, thus its ability to invade the Pantanal may not be surprising, but the invasion of C. americana is counterintuitive, because this species is adapted to the well-drained soils of the cerrado. Thus, we were interested in comparing the photosynthetic capacity, in terms of CO₂ conductance, carboxylation, and electron transport of these species over a seasonal flooding cycle. Given that V. divergens is reportedly flood-adapted, we predicted that this species would have a higher photosynthetic capacity than C. americana, especially under flooding. To test this hypothesis we measured the photosynthetic CO₂ response (P _N/C _c) of V. divergens and C. americana within 1 year to determine, if photosynthetic capacity varied systematically over time and between species. Contrary to our hypothesis, V. divergens did not always have a higher photosynthetic capacity than C. americana. Rather, species differences were influenced by temporal variations in flooding and the leaf age. Leaf CO₂ assimilation and photosynthetic capacity of both species were lower during the flood period, but the differences were not statistically significant. The physiological performance of both species was strongly related to leaf N and P concentrations, but P limitation appeared to be more important than N limitation for these species and ecosystem. Photosynthetic capacity was higher and more stable for V. divergens, but such an advantage did not result in a statistically significant increase in P _N. Our results suggest that both species are tolerant to flooding even though they are adapted to very different hydrological conditions. Such physiological plasticity, especially for C. americana, might be a key feature for the ability to survive and persist in the seasonally flooded Pantanal.     

Neutralising antibodies for West Nile virus in horses from Brazilian Pantanal

Despite evidence of West Nile virus (WNV) activity in Colombia, Venezuela and Argentina, this virus has not been reported in most South American countries. In February 2009, we commenced an investigation for WNV in mosquitoes, horses and caimans from the Pantanal, Central-West Brazil. The sera of 168 horses and 30 caimans were initially tested using a flaviviruses-specific epitope-blocking enzyme-linked immunosorbent assay (blocking ELISA) for the detection of flavivirus-reactive antibodies. The seropositive samples were further tested using a plaque-reduction neutralisation test (PRNT90) for WNV and its most closely-related flaviviruses that circulate in Brazil to confirm the detection of specific virus-neutralising antibodies. Of the 93 (55.4%) blocking ELISA-seropositive horse serum samples, five (3%) were seropositive for WNV, nine (5.4%) were seropositive for St. Louis encephalitis virus, 18 (10.7%) were seropositive for Ilheus virus, three (1.8%) were seropositive for Cacipacore virus and none were seropositive for Rocio virus using PRNT90, with a criteria of ≥ four-fold antibody titre difference. All caimans were negative for flaviviruses-specific antibodies using the blocking ELISA. No virus genome was detected from caiman blood or mosquito samples. The present study is the first report of confirmed serological evidence of WNV activity in Brazil.     

Trypanosoma evansi control and horse mortality in the Brazilian Pantanal

The impact of three treatment strategies for Trypanosoma evansi control on horse mortality in the Brazilian Pantanal based on four size categories of cattle ranches is explored. The region's 49,000 horses are indispensable to traditional extensive cattle ranching and T. evansi kills horses. About 13% of these horses would be lost, annually, due to T. evansi if no control were undertaken. One preventive and two curative treatment strategies are financially justifiable in the Pantanal. The best available technology for the treatment of T. evansi from a horse mortality perspective is the preventive strategy, which spares 6,462 horses, annually. The year-round cure spares 5,783 horses, and the seasonal cure saves 5,204 horses on a regional basis relative to no control strategy. Regardless of the strategy adopted, 39% of the costs or benefits fall to the largest ranches, while 18% fall to the smallest ranches.     

New challenges in the management of the Brazilian Pantanal and catchment area

The Pantanal wetland is a vast seasonally inundated area of extraordinary landscape and biological diversity and complexity. It is located in the upper portion of the Paraguay River basin in central South America. During the rainy season, increased stream discharge from the surrounding basin produces an annual flood pulse through the Pantanal. Increasing human impact, such as dam construction, deforestation, agricultural related activities, and the Hidrovia project in the Parana-Paraguay waterway, threaten the ecological stability of the Pantanal area. As a result, there is an urgent need to introduce new management practices in the Pantanal Basin. In this paper we present a concept for managing the Pantanal catchment based on the integration of ecological knowledge, institutional organization, and involvement of different stakeholders. We propose approaches for an integrated management of the Pantanal and its catchment based on improving data bases and the empowerment of the stakeholder groups. The latter depends on increasing the level of education and access to information, as well as implementing procedures to improve public involvement and enforcement of environmental regulations.This revised version was published online in March 2005 with corrections to the issue cover date.     

Selected oxidative stress markers in a South American crocodilian species

Crocodilians and other diving vertebrates experience hypoperfusion and hypoxia of several internal organs during long dives. At the end of a dive, reperfusion of aerated blood may cause a physiologically relevant oxidative stress. In this study, we analyzed selected markers of oxidative stress in eight organs of normoxic Paraguayan caiman (Caiman yacare) captured in the Brazilian Pantanal wetlands during the winter of 2001 (six mature-adult males and eight young-adult males; AD-1 and YA-1 groups, respectively), and during the summer of 2002 (six young-adult males (YA-2 group), ten hatchlings and five embryos). Lipid peroxidation products determined by three different assays were generally highest in brain, liver and kidney (in comparison with all other organs), and lowest in white muscles from the tail and hind legs. Liver and kidney showed the highest levels of carbonyl protein, while brain showed low levels. Intermediate levels of oxidative stress markers were mostly found in the heart ventricles and lung. Differences in oxidative stress markers between AD-1 and YA-1 were organ-specific, showing no age-related correlation. However, most oxidative stress markers in YA-2 organs were either higher than (by 1.4- to 3.7-fold) or not significantly different from respective values in hatchlings organs. This pattern (hatchlings versus young-adults) was confirmed using correlation analysis of individual caiman size versus levels of oxidative damage markers in four organs. The higher level of oxidative stress markers in young-adults possibly relates to the fast growth rate (and thus, increased oxidative metabolic rate) of C. yacare in the first years of life. Differences in oxidative stress markers between YA-1 and YA-2 were also observed and were ascribed to seasonal changes in free radical metabolism. These results in normoxic C. yacare represent the first step towards understanding the age-related physiological oxidative stress of a diving reptile from a seasonally changing wetland environment.     

Renal antioxidant enzymes and glutathione redox status in leptin-induced hypertension

Previously, we have demonstrated that leptin increases blood pressure (BP) in the rats through two oxidative stress-dependent mechanisms: stimulation of extracellular signal-regulated kinases (ERK) by H(2)O(2) and scavenging of nitric oxide (NO) by superoxide (O(2-.)). Herein, we examined if renal glutathione system and antioxidant enzymes determine the mechanism of prohypertensive effect of leptin. Leptin administered at 0.5 mg/kg/day for 4 or 8 days increased BP and renal Na(+),K(+)-ATPase activity and reduced fractional sodium excretion; these effects were prevented by NADPH oxidase inhibitor, apocynin. Superoxide scavenger, tempol, abolished the effect of leptin on BP and renal Na(+) pump in rats receiving leptin for 8 days, whereas ERK inhibitor, PD98059, was effective in animals treated with leptin for 4 days. Leptin administered for 4 days decreased glutathione (GSH) and increased glutathione disulfide (GSSG) in the kidney. In animals receiving leptin for 8 days GSH returned to normal level, which was accompanied by up-regulation of gamma-glutamylcysteine synthetase (gamma-GCS), a rate-limiting enzyme of the GSH biosynthetic pathway. In addition, superoxide dismutase (SOD) activity was decreased, whereas glutathione peroxidase (GPx) was increased in rats receiving leptin for 8 days. Cotreatment with gamma-GCS inhibitor, buthionine sulfoximine (BSO), accelerated, whereas GSH precursor, N-acetylcysteine (NAC), attenuated leptin-induced changes in gamma-GCS, SOD, and GPx. In addition, coadministration of BSO changed the mechanism of BP elevation from H(2)O(2)-ERK to (O(2-.))-NO dependent in animals receiving leptin for 4 days, whereas NAC had the opposite effect in rats treated with leptin for 8 days. These results suggest that initial change in GSH redox status induces decrease in SOD/GPx ratio, which results in greater amount of (O)2-.)) versus H(2)O(2) in later phase of leptin treatment, thus shifting the mechanism of BP elevation from H(2)O(2)-ERK to (O(2-.))-NO dependent.     

Genetic status of the wood stork (Mycteria americana) from the southeastern United States and the Brazilian Pantanal as revealed by mitochondrial DNA analysis

The wood stork (Mycteria americana) is a colonial wading bird that inhabits the Neotropical region from the southeastern United States (US) to northern Argentina. The species is considered to be endangered in the US due to degradation of its foraging and breeding habitat. In other parts of its range, such as in the Brazilian Pantanal region, breeding populations of this species appear to be stable. We compared the levels of genetic variability and population structuring of the US and the Pantanal breeding populations using mitochondrial DNA (mtDNA) control region sequences. Twenty-seven haplotypes were identified among 88 wood stork samples collected from eight breeding colonies in the US and eight in the Pantanal. Patterns indicative of heteroplasmy were observed in 35.3% of the mtDNA sequences that were examined. Significantly higher levels of haplotype diversity were observed in the Pantanal samples compared to those from the US, suggesting that during the last century, demographic declines or a recent evolutionary bottleneck reduced the levels of mtDNA variability of the US population. Analyses of genetic structuring revealed non-significant genetic differentiation between these regions, indicating that either the populations were only recently separated or that gene flow continues to occur at low levels. Haplotype network analysis indicated low current levels of gene flow between populations that were closely related in the past.     

Using dung beetles to evaluate the conversion effects from native to introduced pasture in the Brazilian Pantanal

The Pantanal is the largest Neotropical seasonal freshwater wetland on Earth. Extensive livestock production has been the dominant economic land use activity of the Pantanal, where approximately 80 % of the land is occupied by native and introduced pastures. However, the impact of native pasture conversion into introduced pasture on the biodiversity of this biome is little understood. Here we evaluate the effect of native pasture to introduced pasture conversion on dung beetle communities. We sampled dung beetles in July 2011 (dry season) and January 2012 (rainy season), at four native pasture sites and four introduced pasture sites in Aquidauana, Mato Grosso do Sul, Brazil. The sampling was carried out using pitfall traps baited with three different bait types: carrion, cattle dung, and human feces. We sampled 7086 individuals, belonging to 32 species of 16 genera and six tribes of dung beetles. The abundance was similar among the pasture types. However, a higher species richness was found on the native pasture. Species composition also differed between the two pasture types in each sampling season. Additionally, the dominant functional guilds were different in the two landscapes. The result shows that the conversion of native grasslands into introduced pasture results in a decrease of species number and changes in species composition. These findings highlight the importance on native pasture to the conservation of dung beetle biodiversity in this ecosystem.     

Spatiotemporal variation in human-wildlife conflicts along highway BR-262 in the Brazilian Pantanal

The Pantanal of Brazil, the world’s largest freshwater wetland, supports a large diversity of species and is under continually increasing pressure from human development, including vehicle collisions with wild animals. We examined decadal, annual, and spatial variation in wildlife-vehicle collisions along highway BR-262 in the southern Pantanal, and specifically: (1) what is the rate of roadkill along BR-262, and did it increased over the previous decade, (2) which species are frequently victims of collisions and does this vary annually along the highway’s length, and (3) what anthropogenic or environmental factors may influence this variation and how can the context of collisions inform mitigation? We sampled the highway monthly, between April 2011 and June 2012, stopping when roadkill was observed to identify species and collect GPS coordinates and a photographic record. Data was analyzed temporally and in relation to traffic volume and precipitation, and imported into a GIS and analyzed spatially and in relation to elevation. We recorded 518 carcasses from 40 species over the 15 sampling periods. The rate of roadkill incidents was 1 every 6.2 km, a tenfold increase since 2002, and is now one of the highest in Brazil. IUCN Red List species were observed, including lowland tapir, giant anteater, and marsh deer. At the lowest elevations, during seasonal wetland inundation the causeway embankment is sought for refuge, increasing collision probability. Mitigation measures tied to seasonal flood pulse variation along the causeway may reduce the high rate of wild animal mortality and loss of biodiversity.     

Ant communities,environmental characteristics and their implications for conservation in the Brazilian Pantanal

Studies about processes determining biodiversity in Pantanal are urgently needed and the use of invertebrates for this purpose may be an efficient tool. The purpose of this paper is to verify whether ants could be used to predict human impacts on Pantanal, assessing how environmental characteristics of gallery forest influence arboreal ant species richness. Specifically, we tested the hypotheses that: (i) resource availability (estimated by tree density) and (ii) diversity of environmental conditions (estimated by structural heterogeneity) positively influence arboreal ant species richness and composition. Ants were sampled in a gallery forest in the Miranda River. Ant species richness did not respond to tree density, but increased with structural heterogeneity. Species composition was affected both by tree density and structural heterogeneity. The species list produced in the present study showed a high similarity between arboreal ant faunas of Cerrado and Pantanal, which are contiguous biomes. The relationship of ant species richness with heterogeneity, as well as the observed changes of species composition with tree density and heterogeneity may be explained by the presence of a variation of environmental conditions, which create several micro-habitats in the forest. The response of the ant community can reflect human impacts on the forests of the studied region, which are usually modified to create patches for sport fishing and by pathways used in ecotourism.     

Changes in the antioxidant status in leaves of Solanum species in response to elicitor from Phytophthora infestans

Three Solanum genotypes with various polygenic resistance levels to the oomycete pathogen Phytophthora infestans (Mont.) De Bary were studied for their antioxidant response to the pathogen culture filtrate (CF). Detached plant leaves were treated with CF for 6, 18 and 30 h, and assayed for changes in hydrogen peroxide content, total ascorbate and glutathione pools and redox ratios (reduced form to total pool), as well as for changes in activities of ascorbate peroxidase, glutathione reductase and glutathione-S-transferase. In CF treated leaves of non-host resistant S. nigrum var. gigantea and field resistant S. tuberosum cv Bzura, the H(2)O(2) content did not change in comparison to water treated control leaves, whereas in the susceptible S. tuberosum clone H-8105 it decreased below the control level. In CF treated leaves of all genotypes, the total ascorbate pools were relatively unaltered and their redox ratio changed only transiently. In Bzura leaves the total glutathione content increased earlier than in the two other genotypes. The glutathione redox ratio remained rather stable, except for the susceptible clone H-8105, where it decreased transiently by about 42%. The relative increases in activity of all the studied enzymes were the highest in the susceptible clone H-8105. The results are discussed in the light of oxidative processes occurring in CF treated leaves. We conclude that stringent control of pro- and anti-oxidant reactions bringing the H(2)O(2) and/or cellular redox state to the threshold level is decisive for deployment of an effective defense strategy.     

Arthropod Biodiversity in the Canopy of Vochysia divergens (Vochysiaceae), a Forest Dominant in the Brazilian Pantanal

The canopy of an invasive tree species in the Pantanal floodplains, Vochysia divergens Pohl (Vochysiaceae; height 12 m) was sprayed once with 5% synthetic pyrethrum during the dry, non-flooded season near Poconé, Mato Grosso. About 4200 arthropods collected (43.7 ind./m 2) represented 14 orders. The Hymenoptera dominated (21.4 ind./ 2 ; mostly ants), followed by Coleoptera, Araneae and Diptera (85% of the total catch). Of the 26 Coleoptera families (6.2 ind./ 2) the Curculionidae, Scolytidae, Elateridae and Dryopidae were most abundant (70% of the total catch), and on trophic level the herbivores dominated (42%). This is the first quantitative canopy study of a tree species in the northern Pantanal floodplains. The type of data needed on arthropods from tree canopies in future studies to aid in the management and conservation of Pantanal floodplains is discussed.     

Generation of a sequence characterized amplified region probe for authentication of crocodilian species

A 209-base pair (bp) crocodilian-specific sequence characterized amplified region (SCAR) was identified from a 425-bp randomly amplified polymorphic DNA (RAPD) fragment. The 209-bp SCAR was produced from amplifications of DNA extracted from fresh and/or dry meat samples from at least three species of Crocodylus, Caiman crocodylus, and Alligator mississippiensis. No amplification was observed from DNA of other common animal species. The use of SCAR opens the way for quick authentication of crocodilian samples for conservation biology and trade regulation.     

New Stenella and Parastenella species from the Brazilian cerrado

Five new Stenella species were found on native cerrado plants. Stenella erythroxyli-campestris, S. erythroxyli-suberosi and S. erythroxylicola were associated with plant species belonging in the family Erythroxylaceae; S. cyrtopodii was found infecting the rare Cyrtopodium eugenii (Orchidaceae), and S. ocoteae occurred on Ocotea sp. (Lauraceae). Finally Parastenella callisthenis-fasciculatae was collected on a Vochysiaceae (viz. Callisthene fasciculate) endemic to the cerrado.     

Glutathione and glutathione metabolizing enzymes in yeasts

Total glutathione content, glutathione peroxidase, glutathione transferase and glutathione reductase activities have been measured in 12 species of yeasts. All the strains tested contained glutathione, though in different amounts, as well as the above mentioned enzymes. To discriminate between the selenium-dependent and the selenium-independent form, glutathione peroxidase activity has been measured with both H₂O₂ and cumene hydroperoxide. Rhodotorula glutinis appeared to be the only strain in which the selenium-dependent form was not found, but this yeast exhibited the highest level of selenium-independent glutathione peroxidase activity as compared to the other strains.     

Effect of reactive oxygen and carbonyl species on crucial cellular antioxidant enzymes

Numerous reactive oxygen species (ROS) and reactive carbonyl species (RCS) issuing from lipid and sugar oxidation are known to damage a large number of proteins leading to enzyme inhibition and alteration of cellular functions. Whereas studies in literature only focus on the reactivity of one or two of these compounds, we aimed at comparing in the same conditions of incubations (4 and 24 h at 37 °C) the effects of both various RCS (4-hydroxynonenal, 4-hydroxyhexenal, acrolein, methylglyoxal, glyoxal, malondialdehyde) and ROS (H₂O₂, AAPH) on the activity of key enzymes involved in cellular oxidative stress: superoxide dismutase (Cu,Zn-SOD), glutathione peroxidase (GPx), glutathione S-transferase (GST) and glucose-6-phosphate dehydrogenase (G6PDH). This was realized both in vitro on purified proteins and MIAPaCa-2 cells. Incubation of these enzymes with RCS resulted in a significant time- and concentration-dependent inhibition for both pure enzymes and in cell lysates. Among all RCS and ROS, hydroxynonenal (HNE) was observed as the most toxic for all studied enzymes except for SOD and is followed by hydrogen peroxide. At 100 μM, HNE resulted in a 50% reduction of GPx, 56% of GST, 65% of G6PDH, and only 10% of Cu,Zn-SOD. Meanwhile it seems that concentrations used in our study are closer to biological conditions for ROS than for RCS. H₂O₂ and AAPH-induced peroxyl radicals may be probably more toxic towards the studied enzymes in vivo.     

Mercury accumulation in sediment cores and along food chains in two regions of the Brazilian Pantanal

The Pantanal is a 140,000 km² floodplain wetland stretching acrosswestern Brazil and parts of Bolivia and Paraguay. Gold mining withmercury (Hg) amalgamation has thrived since 1980 along its northern rim. We quantified Hg accumulation in sediment cores (N = 5) and food chainsin this general region of the northern Pantanal and in a reference region,200 km deeper into the wetland (Acurizal). Cores were dated with²¹⁰Pb and ¹³⁷Cs using direct gamma-assay. Total Hg wasanalyzed by cold-vapor atomic fluorescence using a gold-meshpre-concentration trap. Average pre-1940 Hg accumulation in cores wasnot significantly different (N = 5, p= 0.14) between both regions andcomparable with rates calculated for global reference sites. Post gold-rushHg (post-1980) deposition averaged 55 ± 11.3 g m^-2yr^-1 in the northern impacted region and was more than 1.5 timeshigher than the post-1980 rate in Acurizal, implying a regional Hg effectof gold mining. Post-1980 Hg accumulation in Acurizal, in turn, was 2.1times the rate reported for a global reference during that time period,suggesting an additional basin-wide effect over such reference sites. Bycombining our core data with assessments of the size of the impacted areaand the amount of Hg released to the region since 1980, we estimated thatonly 2–8% of this Hg was recovered as a sedimentary signal. Theremainder of the Hg was lost to the atmosphere, downstream areas, orstored in biota. Hg concentrations in surface sediments in the northernPantanal (45.5 ± 5.5 ng g_dry ^-1) were significantlyhigher than those in our reference region (29.1 ± 0.7ng g_dry ^-1). Hg levels in primary producers were alsoelevated in the northern Pantanal. Eichhornia crassipes rootscontained 2.7–3.0 times more mercury than shoots in both regions and Salvinia auriculata, suggested as a biological monitor for Hg pollution,contained almost four times more mercury in the northern Pantanal (90.7± 9.1 ng g_dry ^-1) than in Acurizal (24.5 ± 3.3ng g_dry ^-1). Plant grazers and scavengers, such as apple snails(Pomacea sp.) and adult water beetles (Fam. Hydrophilidae), werelow in Hg, confirming previous data showing that the channeling of mercuryfrom lower to higher trophic levels along herbivorous links was inefficientcompared to transfer along carnivorous links. Collections of 12–16individuals of four species of Characidae (Aphyocharax sp., Tetragonopterus sp., Serrasalmus spiropleura and Pygocentrisnattereri) in both regions showed elevated Hg body burdens in bothpiranhas S. spiropleura and P. nattereri from the northernPantanal (149.9 ± 84.2 and 302.2 ± 159.1ng g_dry ^-1, respectively). Fish length for each species was notstatistically different between regions. P. nattereri length correlatedsignificantly (p<0.001) with Hg content in both regions, but the slopeof the regression in the northern Pantanal was 2.6 times the slope for theAcurizal collection, indicating an elevated rate of biomagnification in theHg-impacted region. Signals of Hg use in mining can be quantified insediment core chronologies and biological tissues, although species atdifferent trophic levels show dissimilar impacts. Mechanisms involved in Hgmagnification along food chains deserve more attention, particularly intropical regions where the threat of chronic exposure to this neurotoxinmay have the greatest implications for biodiversity.