Ecological factors associated with European bat lyssavirus seroprevalence in spanish bats.

Bats have been proposed as major reservoirs for diverse emerging infectious viral diseases, with rabies being the best known in Europe. However, studies exploring the ecological interaction between lyssaviruses and their natural hosts are scarce. This study completes our active surveillance work on Spanish bat colonies that began in 1992. Herein, we analyzed ecological factors that might affect the infection dynamics observed in those colonies. Between 2001 and 2011, we collected and tested 2,393 blood samples and 45 dead bats from 25 localities and 20 bat species. The results for dead confirmed the presence of EBLV-1 RNA in six species analyzed (for the first time in Myotis capaccinii). Samples positive for European bat lyssavirus-1 (EBLV-1)-neutralizing antibodies were detected in 68% of the localities sampled and in 13 bat species, seven of which were found for the first time (even in Myotis daubentonii, a species to date always linked to EBLV-2). EBLV-1 seroprevalence (20.7%) ranged between 11.1 and 40.2% among bat species and seasonal variation was observed, with significantly higher antibody prevalence in summer (July). EBLV-1 seroprevalence was significantly associated with colony size and species richness. Higher seroprevalence percentages were found in large multispecific colonies, suggesting that intra- and interspecific contacts are major risk factors for EBLV-1 transmission in bat colonies. Although bat-roosting behavior strongly determines EBLV-1 variability, we also found some evidence that bat phylogeny might be involved in bat-species seroprevalence. The results of this study highlight the importance of life history and roost ecology in understanding EBLV-1-prevalence patterns in bat colonies and also provide useful information for public health officials.

Trace metal concentrations in tissues of two tinamou species in mining areas of Bolivia and their potential as environmental sentinels.

Mining has a long history in the Bolivian Andes and has left many tailing piles, from which trace metals may reach surface waters, soils, and biota. The potential of tinamous (Birds: Tinamidae) as sentinels has never been tested before, although their biological and ecological characteristics mean they could well be appropriate bioindicators. We captured 13 and nine individuals of the Ornate Tinamou (Nothoprocta ornata) from two polluted sites (P1 and P2) and 10 and five from control unpolluted sites (NP1 and NP2) and used, for comparative purposes, four specimens bred in captivity. We also captured six specimens of Darwin's Nothura (Nothura darwinii) from the polluted site, P2. We determined the concentration of As, Cd, Pb, and Sb in feathers, liver, and kidney and conducted histological analyses of liver and kidney. For the Ornate Tinamou, a site effect was found for all trace metals in all tissues, with the highest concentrations at polluted sites. At the P2 site, no differences between the two tinamou species were detected except in some cases where Darwin's Nothura shows near-double concentrations. In some cases, mean and/or individual values of trace metal concentrations reached toxicity levels at the polluted sites. Thesaurismosis in proximal convoluted renal tubules, probably related to Cd exposure, was observed in 30% of the samples from the P1 site. Significant correlations were observed between all tissues for all trace metals and also for all trace metals in each tissue. Because the species studied are ubiquitous and relatively abundant, we recommend monitoring programs based on feather analysis.

Bioaccumulation of metals and effects of a landfill in small mammals Part III: Structural alterations.

The leachates from the Garraf landfill located in a protected site (NE Spain) contain several potentially toxic substances such as heavy metals. Here we report the histopathological alterations produced by this pollution in wild specimens of an omnivorous species, the wood mouse, Apodemus sylvaticus, and an insectivorous species, the greater white-toothed shrew, Crocidura russula. Hepatic tissue presented the most severe alterations in both the species, namely cell cycle arrest (apoptosis and necrosis), inflammation, preneoplasic nodules, vacuolation and microsteatosis. The kidneys were altered more in the mice (presenting tubular necrosis and dilatation, inflammation, and cylinders) than in the shrews, suggesting that different metabolic pathways render shrews more tolerant to renal toxicity induced by pollutants. No pollution-related alterations were observed in lung, spleen, pancreas, gonads, oesophagus, intestine, or adrenals. We conclude that the two species could be used in conjunction as bioindicators to assess the effects of environmental pollution at different trophic levels.

Metals in liver and kidneys and the effects of chronic exposure to pyrite mine pollution in the shrew Crocidura russula inhabiting the protected wetland of Doñana.

Historically impacted by anthropogenic activities, the nature reserve of Doñana (SW Spain) was affected by an unprecedented spillage of mud and acidic water from the Aznalcóllar pyrite mine in April 1998. Although several studies have addressed the influence of this spill on soils, water, and biota, there is little information on mammals, especially carnivorous species. We measured the concentrations of Fe, Mg, Pb, Hg, Cd, Zn, Cu, Mn, Mo, Co, and Cr in specimens of the greater white-toothed shrew, Crocidura russula, inhabiting the protected area affected by the mine spillage. We also examined other parameters to approach at the physiological effects of pollution. We found an increase in non-essential metals (Pb, Cd, and Hg), and morphometric, histological and genotoxic alterations. Age and gender were two significant factors explaining metal bioaccumulation: adults had higher Hg and Cd levels than juveniles, whereas males bioaccumulated more Pb and Co and less Mo than females. The micronucleus frequencies in blood erythrocytes were significantly higher in specimens from the polluted site than animals from the control site. Shrews from the impacted area also had hepatic alterations, namely increased liver-body ratio, focal necrosis, and signs of apoptosis in hepatocytes. Due to the relevance of small mammals in the diet of endangered species such as carnivorous birds and mammals, the findings of our study are of practical use for the management of the Doñana wildlife reserve and other protected Mediterranean wetlands.

Bioaccumulation of metals and effects of a landfill in small mammals. Part II. The wood mouse, Apodemus sylvaticus.

We assess the bioaccumulation of metals (Pb, Hg, Cd, Fe, Mg, Zn, Cu, Mn, Mo, Cr) and effects of landfill leachates on morphological (RI, relative weights), plasma (GPT, GOT, creatinine), and genotoxic (MNT) parameters in wood mice, Apodemus sylvaticus, inhabiting close the Garraf landfill site (NE Spain). Due to the high age- and sex-dependent variation in wild populations, we also studied the effect of these biotic factors on the parameters studied. Wood mice from the landfill site, sited in a partially protected area, showed more Cd, Fe, Zn, Cu, Mn, Mo, and Cr than specimens from the reference site. Moreover, mice near the landfill registered low RI and high relative renal weight, GPT, and MN frequency, which indicate that the landfill affects the health of wild mice. In contrast to sympatric shrews from a previous study, wood mice showed lower bioaccumulation of metals and lower variation caused by biotic factors. Moreover, the morphological and physiological alterations demonstrated that they were also more sensitive at environmental pollution. Given the contribution of small mammals to ecosystem function and the scarce ecotoxicological data on the effects of landfill pollution on wild terrestrial mammals, we consider that our study can be used to improve the management of this protected area.

Bioaccumulation of metals and effects of landfill pollution in small mammals. Part I. The greater white-toothed shrew, Crocidura russula.

Here we quantified the bioaccumulation of metals (lead, mercury, cadmium, iron, magnesium, zinc, copper, manganese, molybdenum, and chromium) and assessed several morphological (RI, relative weights) and genotoxic parameters as biomarkers of pollution from the landfill of Garraf (Barcelona, NE Spain). Specimens of Crocidura russula (Insectivora, Mammalia) from the landfill site showed increased Pb, Cd, Mg, Zn, Cu, and Cr concentrations in their tissues. Levels of mercury were below detection limits. Concentrations of Cd, Pb, and Cr varied significantly with age and no differences were found between males and females. While no differences were found in morphological parameters between shrews from the two sites, those from the polluted one showed more micronuclei in blood than those from reference site (1.786+/-0.272 vs. 0.088+/-0.045 per thousand; U=46.000, p<0.001). The considerable amounts of potentially toxic metals (Pb till 59.71 and Cd till 56.57microgg(-1) DW in kidneys) and the genotoxic effects indicate the harmful effect on biota. We consider necessary biomonitoring this landfill sited in a partially protected area.

How does the greater white-toothed shrew, Crocidura russula, responds to long-term heavy metal contamination? -- A case study.

Heavy metals accumulation in parallel with the evaluation of physiological and biochemical effects resulting from continued metal exposure were considered here using for the first time the great white-toothed shrew Crocidura russula as an in vivo model. Shrews were originated from an abandoned lead/zinc mining area and from a reference area, both in Alentejo, southern Portugal. Hepatic contents of nickel, copper, zinc, cadmium, mercury and lead were quantified by Inductively Coupled Plasma Mass Spectrometer (ICP-MS). Haematological parameters (white blood cells, red blood cells, haemoglobin and haematocrit) were obtained in a Coulter Counter Analyser and biochemical markers of the redox balance (glutathione S-transferase, glutathione peroxidase, and glutathione reductase) activities were measured spectrophotometrically using a Duo-50 spectrophotometer. Compared with control animals, significantly higher concentration of hepatic cadmium (9.29 vs. 1.18 micorg/g dry weight) and nickel (1.56 vs. 0.343 microg/g dry weight) were detected in the shrews collected in the mining area. However, no significant changes were observed on haematological or enzymatic parameters in animals exposed to metal pollution. The obtained results show that shrews are good bioaccumulators of toxic heavy metals, but very tolerant to their effects, revealing an interesting long-term adaptation to polluted environments. In addition, this study provides reference values for haematological parameters and antioxidant enzymes levels in C. russula, which may be relevant for comparative purposes in further studies.

Bioaccumulation of lead, mercury, and cadmium in the greater white-toothed shrew, Crocidura russula, from the Ebro Delta (NE Spain): sex- and age-dependent variation.

We quantified bioaccumulation of lead, mercury, and cadmium in bones from 105 greater white-toothed shrews (Crocidura russula) collected at the Ebro Delta, a polluted area, and the Medas Islands, a control site. Lead and mercury levels varied with site, age, and sex, although statistical significances depended on each factor. Globally, shrews from the polluted area exhibited significantly higher concentrations of Pb and Hg. Increment of Pb with age was particularly remarkable in wetland animals and was interpreted in relation to human activities, namely hunting. Unlike males, females from the Ebro Delta maintained low Hg levels, which were associated with gestation and lactation. Cadmium levels did not differ between sites, sexes, or ages. This study provides the first data on heavy metals in mammals from this wetland and suggests that C. russula is a good bioindicator of metal pollution. We concluded that sex and age may represent an important source of variation in the bioaccumulation of these metals in wild populations.

Metal bioaccumulation in the greater white-toothed shrew, Crocidura russula, inhabiting an abandoned pyrite mine site.

Hepatic and renal concentrations of iron, magnesium, zinc, lead, copper, manganese, mercury, cadmium, molybdenum, chromium, and nickel were quantified in shrews (Crocidura russula) inhabiting a pyrite mine site in Portugal. Several morphological parameters (body weight, residual index, and relative weights) were also examined to clarify the physiological effects of pollution. Shrews from the mine showed increased bioavailability of Fe, Pb, Hg, Cd, Mo, and Ni in comparison with reference specimens. Adult shrews had the highest Cd levels while Cr and Ni concentrations diminished. Intersexual differences were found for Mo and Ni. As a consequence of metal pollution, the relative hepatic weight was higher in shrews from the mine site when compared with reference specimens. These data indicate that C. russula is a good bioindicator of metal pollution. We also evaluated the toxic effects of Pb, Hg, Cd, and Ni, because several shrews from the polluted site showed high concentrations of these metals. To approximate at the real biological impact of abandoned mines, after this first step it is necessary to associate the bioaccumulation levels and morphological effects with other physiological, ecological and genetical biomarkers.