Mopeia virus-related arenavirus in natal multimammate mice, Morogoro, Tanzania.

A serosurvey involving 2,520 small mammals from Tanzania identified a hot spot of arenavirus circulation in Morogoro. Molecular screening detected a new arenavirus in Natal multimammate mice (Mastomys natalensis), Morogoro virus, related to Mopeia virus. Only a small percentage of mice carry Morogoro virus, although a large proportion shows specific antibodies.

Do wood mice (Apodemus sylvaticus L.) use food selection as a means to reduce heavy metal intake?

Food preference of wood mice from two with heavy metals polluted sites and two unpolluted sites was tested under laboratory and field conditions with two-way choice experiments. In the laboratory, wood mice preferred to eat acorns from unpolluted sites over acorns from polluted sites. Previous experience with polluted food had no influence on food choice. Preference was negatively related to acorn metal content. Furthermore, the nutrient content of the acorn endosperm was consistently lower in polluted sites. We therefore conclude that wood mice used absolute metal concentration in the acorn, nutrient content, or both as a food selection cue. The results of the laboratory experiment could not be confirmed under field conditions. We hypothesized that search time constraints due to the presence of predators, competitors and/or other stress factors in the field have prevented the mice to forage selectively.

Non-destructive pollution exposure assessment by means of wood mice hair.

Concentrations of cadmium, copper, lead and zinc were measured in hair, kidney, liver, lung and muscle tissue of wood mice captured along a pollution gradient. We found positive relationships between cadmium concentrations in hair and all internal tissues. Hair lead concentrations were positively correlated with lead contents in kidney and liver. Age had a significant effect on cadmium accumulation in all tissues and hair. Apart from a very weak relationship between zinc concentrations in hair and liver, no significant relation between copper or zinc content in hair and any of the internal organs was observed. In summary, our observations suggest that hair of wood mice can be used for monitoring exposure to non-essential metals like cadmium and lead, but not to homeostatically regulated metals such as copper or zinc.

Non-destructive pollution exposure assessment in the European hedgehog (Erinaceus europaeus): IV. Hair versus soil analysis in exposure and risk assessment of organochlorine compounds.

Few ecotoxicological studies on mammals use non-destructive methodologies, despite the growing ethical concern over the use of destructive sampling methods. In the present study we assessed exposure of hedgehogs (Erinaceus europaeus) to polychlorinated biphenyls (PCBs), dichloro-diphenyl-trichloroethanes (DDTs), hexachlorocyclohexanes (HCHs) and hexachlorobenzene (HCB), by investigating concentrations of these compounds in soils and hedgehog hair from seven study sites around the urban area of Antwerp, Belgium. No relationships were observed between organochlorine compound concentrations in soils and hair from the different study areas. Furthermore, the individual variation of contamination levels in hair within study sites was high, especially for HCHs and HCB, and hair and soil had different relative profiles for PCBs, DDTs and HCHs. Our results show that concentrations of organochlorine compounds in soils alone are not predictive of the risk of these pollutants to hedgehogs and that tissue analyses are preferred to soil analyses in exposure and risk assessment studies.

Spatial spreading of Echinococcus multilocularis in Red foxes (Vulpes vulpes) across nation borders in Western Europe.

The occurrence of the fox tapeworm Echinococcus multilocularis in Red foxes was studied in Belgium and a neighbouring region in The Netherlands. A total number of 1202 foxes were analysed (1018 in Belgium and 184 in The Netherlands) of which 179 were infected with E. multilocularis (164 in Belgium and 15 in The Netherlands). Further, the spatial distribution of infection among sampled foxes was analysed with an ellipsoidal gradient, demonstrating a decreasing prevalence in northwestern direction. Using this gradient, we showed that the spatial patterns of infection in Belgium and the neighbouring region in The Netherlands correspond, indicating a continuous distribution of E. multilocularis across the nation borders. Part of the Belgian data allowed investigating temporal changes in the spatial distribution of E. multilocularis. This revealed a northwestern spread of E. multilocularis.

Non-destructive pollution exposure assessment in the European hedgehog (Erinaceus europaeus): II. Hair and spines as indicators of endogenous metal and As concentrations.

The role of hair and spines of the European hedgehog as non-destructive monitoring tools of metal (Ag, Al, Cd, Co, Cr, Cu, Fe, Ni, Pb, Zn) and As pollution in terrestrial ecosystems was investigated. Our results showed that mean pollution levels of a random sample of hedgehogs in Flanders are low to moderate. Yet, individual hedgehogs may be at risk for metal toxicity. Tissue distribution analyses (hair, spines, liver, kidney, muscle and fat tissue) indicated that metals and As may reach considerable concentrations in external tissues, such as hair and spines. Positive relationships were observed between concentrations in hair and those in liver, kidney and muscle for Al, Co, Cr, Cu, and Pb (0.43 < r < 0.85). Spine concentrations were positively related to liver, kidney and muscle concentrations for Cd, Co, Cr, Cu and Pb (0.37 < r < 0.62). Hair Ag, As, Fe and Zn and spine Ag, Al, As and Fe were related to metal concentrations in one or two of the investigated internal tissues (0.31 < r < 0.45). The regression models presented here may be used to predict metal and As concentrations in internal tissues of hedgehogs when concentrations in hair or spines are available. The present study demonstrated the possibility of using hair and spines for non-destructive monitoring of metal and As pollution in hedgehogs.

Metal blood levels and hematological characteristics in wood mice (Apodemus sylvaticus L.) along a metal pollution gradient.

The present study evaluated whether blood could be used as a nondestructive tool for monitoring metal exposure and related hematological effects in wood mice (Apodemus sylvaticus L.) living along a metal pollution gradient. Soil concentrations of arsenic, cadmium, copper, lead, silver, and zinc decreased with distance from the emission source. Blood levels of cadmium and lead differed significantly among sites, whereas those of the other metals did not. Blood levels of cadmium and lead correlated with soil concentrations of cadmium and lead, respectively. No such significant relationships were found for the other measured metals. Hematocrit levels decreased in wood mice from the most polluted site (45.96% +/- 0.53% [mean +/- standard error]) compared to the reference site (48.04% +/- 0.47%). A negative correlation between hematocrit and blood levels of cadmium and lead was found. Erythrocyte count, leukocyte count, hemoglobin concentration, mean corpuscular hemoglobin (average wt of hemoglobin in a red blood cell in pg), and lysozyme activity did not differ among study sites. Mean corpuscular volume (average volume of a red blood cell in microm3) and mean corpuscular hemoglobin concentration (average proportion of hemoglobin in a red blood cell as a %) differed among study sites but showed no relationship with metal exposure. We conclude that whole blood from mice can be used for nondestructive monitoring of the exposure to nonessential metals under natural conditions. The present results indicate that decreased hematocrit levels may be an early warning signal for a negative impact of metal exposure on the oxygen-transport capacity of blood in wood mice in their natural environment.

Nondestructive pollution exposure assessment in the European hedgehog (Erinaceus europaeus): III. Hair as an indicator of endogenous organochlorine compound concentrations.

Concentrations of organochlorine persistent pollutants were investigated in tissues of the European hedgehog (Erinaceus europaeus). Road kills and carcasses from wildlife rescue centers were used to characterize organochlorine compound tissue distribution and tissue profile dissimilarities (hair, liver, kidney, muscle, and adipose tissue). The most important contaminants were polychlorinated biphenyls (PCBs), chlordanes (CHLs), and DDTs, with median concentrations of 75, 5.1, and 1.4 ng/g liver wet weight, respectively. Median levels for the remaining compounds-hexachlorobenzene (HCB), hexachlorocyclohexanes (HCHs), and octachlorostyrene-were less than 0.5 ng/g liver wet weight. Compared to results with other mammalian wildlife, the present results indicate that hedgehogs may accumulate considerable concentrations of organochlorine compounds. Polychlorinated biphenyls and HCB preferably accumulated in liver and muscle tissue. Concentrations of DDTs and HCHs were highest in muscle tissue and hair, respectively. Octachlorostyrene and CHL levels were predominant in liver. The observed positive relationships between concentrations in hair and internal tissues for PCBs, DDTs, HCB, HCHs, and CHLs (0.49 < r < 0.91) and for several individual PCB, DDT, and CHL compounds (0.31 < r < 0.76) indicate the usefulness of hair as a biomonitoring tool of organochlorine compounds. Multivariate profile analyses revealed a higher dominance of less-persistent compounds (p,p'-DDT, alpha-HCH, and PCBs 95, 101, and 149) in hair compared to internal tissues. The present study demonstrates the suitability of hedgehog hair as a nondestructive biomonitoring tool regarding pollution with organochlorine compounds and the promising role of the hedgehog as a mammalian indicator species of pollution in terrestrial environments.

Genotoxicity in wood mice (Apodemus sylvaticus) along a pollution gradient: exposure-, age-, and gender-related effects.

We investigated the effects of environmental pollution on genetic damage in wood mice (Apodemus sylvaticus) by means of the comet assay, with special attention to the role of age and gender as potential confounding variables. The present study was carried out at four sites along a pollution gradient in the vicinity of Antwerp (Belgium), with a nonferrous smelter as the main pollution source. We measured the concentration of heavy metals (Cd, Co, Cr, Cu, Fe, Mn, Pb, and Zn) in mouse liver and kidney and the concentration of organochlorine compounds (polychlorinated biphenyls and 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene) in mouse muscle tissue to assess individual exposure. Cadmium exposure was very high at the sites closest to the smelter, and exposure to this metal decreased with increasing distance from the smelter. Exposure to the other pollutants was low to moderate at the different sites. Genetic damage was higher in mice from populations in the vicinity of the nonferrous smelter compared with that in the control populations. A significant increase in genetic damage with age was observed at the most polluted sites, but not at the control sites. Genetic damage was higher in male mice than in female mice at the most polluted site, but not at the other areas. Yet, no obvious relationship was found between individual pollutant levels and individual genetic damage levels. We conclude that the comet assay can be used to compare genotoxicity at the population level if the confounding variables of gender and age are taken into account. However, its use for individual health risk assessment remains questionable.

Effects of environmental pollution on microsatellite DNA diversity in wood mouse (Apodemus sylvaticus) populations.

Ten microsatellite DNA loci were surveyed to investigate the effects of heavy metal pollution on the genetic diversity and population genetic structure of seven wood mouse (Apodemus sylvaticus) populations along a heavy metal pollution gradient away from a nonferrous smelter in the south of Antwerp (Flanders, Belgium). Analysis of soil heavy metal concentrations showed that soil Ag, As, Cd, Cu, and Pb decreased with increasing distance from the smelter. Genetic analyses revealed high levels of genetic variation in all populations, but populations from the most polluted sites in the gradient did not differ from those of less-polluted sites in terms of mean observed and expected heterozygosity level and mean allelic richness. No correlation was found between measures of genetic diversity and the degree of heavy metal pollution. However, an analysis of molecular variance and a neighbor-joining tree suggested a contamination-related pattern of genetic structuring between the most polluted and less polluted sites. Pairwise F(ST) values indicated that populations were significantly genetically differentiated, and assignment tests and direct estimates of recent migration rates suggested restricted gene flow among populations. Additionally, genetic differentiation increased significantly with geographical distance, which is consistent with an isolation-by-distance model. We conclude that, at least for our microsatellite DNA markers, genetic diversity in the studied wood mouse populations is not affected greatly by the heavy metal pollution.

Nondestructive pollution exposure assessment in the European hedgehog (Erinaceus europaeus): I. Relationships between concentrations of metals and arsenic in hair, spines, and soil.

Conventional metal exposure assessment in terrestrial mammals is generally based on organ analyses of sacrificed animals. Few studies on mammals use nondestructive methodologies despite the growing ethical concern over the use of destructive sampling. Nondestructive methods involve minimal stress to populations and permit successive biomonitoring of the same populations and individuals. In the present study we assessed metal exposure of hedgehogs (Erinaceus europaeus) by investigating relationships between concentrations of metals (Ag, Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Zn) and As in soil samples and in hair and spines of hedgehogs. Samples were collected in seven study sites along a metal pollution gradient, characterized by decreasing total soil Ag, As, Cd, Cu, Ni, and Pb concentrations with increasing distance from a nonferrous metallurgic factory. For a number of elements, soil contamination was related both to distance to the smelter and to habitat. Soil concentrations were positively related to levels in hair and spines for Ag, As, Cd, and Pb and thus to hedgehog exposure. Metal concentrations in soil did not relate to metal concentrations in hair and spines for essential elements (e.g., Cu, Fe, Mn, Ni, and Zn), except Co in hair and soil. Our results demonstrate that, at least for nonessential elements, concentrations in soils can be used to predict contamination of these elements in hedgehogs or vice versa. Furthermore, hedgehog exposure increased toward the smelter and was higher for hedgehogs foraging in grasslands than for animals foraging in the forest. Moreover, we believe that hair and spines are promising tools in terrestrial wildlife exposure assessment studies of metals and As.

RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) AND PARSIMONY METHODS.

- Random amplified polymorphic DNA (RAPD) data possess a number of undesirable features for parsimony analysis. These features include their non-codominant inheritance, their anonymous nature, their different (a)symmetrical transformation probabilities, and their possible GC priming bias. As a consequence, no single parsimony method seems appropriate for RAPD data. Moreover, the presence/absence coding of RAPDs is equivalent to the invalid independent allele model for allozymes. These issues are discussed and the way in which parsimony analysis of RAPDs can be misleading is illustrated.

Nutrient stress, host plant quality and herbivore performance of a leaf-mining fly on grass.

Environmental stresses affect plant growth and performance in nature. Host plant quality in turn affects herbivore performance and population dynamics. In view of these interactions, two major hypotheses were formulated. The plant stress hypothesis proposes that physiologically stressed plants become more susceptible to herbivores. The plant vigour hypothesis proposes that plants that grow vigorously are favourable to herbivores. Here we test the plant stress/plant vigour hypotheses for a leaf miner, Agromyza nigripes (Diptera; Agromyzidae), on the grass Holcus lanatus. We assessed larval performance (survival, developmental time, pupal mass) on grasses growing under different levels of nutrients (Hoagland solution) and drought stress, under controlled field and greenhouse conditions. Plant vigour and nutrient content were high on soils with an intermediate nutrient concentration and lower under drought stress and soil nutrient shortage and overdose. Larval performance was also highest on wet soils with intermediate nutrient supply. The results of the mining flies support the plant vigour hypothesis (density, survival and development better on vigorous plants). Herbivore performance is higher on leaves with a higher protein content.

Lethal and sublethal costs of autotomy and predator presence in damselfly larvae.

We studied the costs of lamellae autotomy with respect to growth and survival of Lestes sponsa damselfly larvae in field experiments. We manipulated predation risk by Aeshna cyanea dragonfly larvae and lamellae status of L. sponsa larvae in field enclosures and compared differences in numbers, size and mass of survivors among treatments. In the absence of a free-ranging A. cyanea larva, about 29% of the L. sponsa larvae died. This was probably due to cannibalism. The presence of a free-ranging A. cyanea reduced larval survival by 68% compared to treatments in which it was absent or not permitted to forage on L. sponsa damselflies. Across all predator treatments, lamellae autotomy reduced survival by about 20%. The mean head width and mass of survivors was lower in the enclosures with a free-ranging A. cyanea compared to the other two predator treatments. This suggested that larvae grew less in the presence of a free-ranging predator, indicating that increased antipredator behaviours were more important in shaping growth responses than reduced population density. Mass, but not head width, of survivors was also reduced after autotomy. The fitness consequences of these effects for the adults may be pronounced. In general, these field data strongly suggest that lamellae autotomy affects population regulation of damselflies.

Immunotoxicology in wood mice along a heavy metal pollution gradient.

We carried out an immunotoxicological field study of wood mice in three populations along a heavy metal pollution gradient. Heavy metal concentrations in liver tissue indicated that exposure to silver, arsenic, cadmium, cobalt and lead decreased with increasing distance from a non-ferrous smelter. Host resistance to the endoparasite Heligmosomoides polygyrus decreased with increasing exposure, while the abundance of tick larvae and the nematode Syphacia stroma was unrelated to heavy metal exposure. Spleen mass was increased at the intermediate and the most polluted sites and was positively correlated with the number of H. polygyrus and tick larvae. Proportion of early apoptotic leukocytes increased towards the smelter and was positively related to cadmium exposure. Red and white blood cell counts and lysozyme activity showed no relationship with metal exposure. All together, our observations suggest negative effects of heavy metal exposure on the immune function of wood mice under field conditions.

Impact of Puumala virus infection on maturation and survival in bank voles: a capture-mark-recapture analysis.

Many zoonotic diseases are caused by rodent-borne viruses. Major fluctuations in the transmission of these viruses have been related to large changes in reservoir host population numbers due to external factors. However, the impact of the pathogen itself on the demography of its reservoir host is often overlooked. We investigated the impact of Puumala virus (PUUV) on survival and reproductive maturation probability of its reservoir host, the bank vole (Myodes glareolus). Three years (2004-06) of data from nine independent sites in southern Belgium were collected and analyzed with a capture-mark-recapture (CMR) method that includes statistical correction for the variation in capture probability of voles. A multistate model based on four states of reproductive activity and PUUV immunoglobulin G (IgG) antibody status was used to estimate survival and probability of transition from one reproductive or infection state to another. Although survival estimates for reproductively active voles were similar between infected and noninfected individuals, PUUV infection in reproductively inactive voles decreased mean monthly survival by 14%. PUUV infection was associated with a threefold increase in the probability of reproductive maturation in bank voles. Moreover, the probability of PUUV IgG seroconversion was three times higher for reproductively active voles compared to reproductively inactive voles. Our model indicates that PUUV infection may alter bank vole population dynamics by affecting both survival and maturation in its host. Additional studies, using CMR methodology with shorter time intervals between trapping sessions and possibly a longer duration, are needed to confirm these findings.

Temporal variation in individual factors associated with hantavirus infection in bank voles during an epizootic: implications for Puumala virus transmission dynamics.

Puumala virus (PUUV), the causal agent of nephropathia epidemica in humans, is one of the many hantaviruses included in the list of emerging pathogens. Hantavirus infection is not distributed evenly among PUUV reservoir hosts (i.e., bank voles [Myodes glareolus]). Besides environmental factors and local population features, individual characteristics play an important role in vole PUUV infection risk. Identifying the relative importance of these individual characteristics can provide crucial information on PUUV transmission processes. In the present study, bank voles were monitored during the nephropathia epidemica outbreak of 2005 in Belgium. Vole sera were tested for presence of immunoglobulin G against PUUV, and a logistic mixed model was built to investigate the temporal variation in individual characteristics and their relative importance to PUUV infection risk in bank voles. Relative risk calculations for individual vole characteristics related to PUUV infection in the reservoir host show that reproductive activity dominates infection risk. The gender effect is only found in reproductively active voles, where reproductively active males have the highest infection risk. Results also revealed a clear seasonal variation in the importance of reproductive activity linked to PUUV infection. In contrast to the main effect found in other trapping sessions, no difference in infection risk ratio was found between reproductively active and nonactive voles in the spring period. Combined with increased infection risk for the reproductively nonactive group at that time, these results indicate a shift in the transmission process due to changes in bank vole behavior, physiology, or climate conditions. Hence, our results suggest that mathematical models should take into account seasonal shifts in transmission mechanisms. When these results are combined with the seasonal changes in population structure during the epizootic period, we identify vole reproductive activity and length of the breeding season as potential drivers of PUUV epizootics in west-central European regions.

Demography, reproductive biology and diet of the bushveld gerbil Tatera leucogaster (Rodentia: Gerbillinae) in the Lake Rukwa valley, south-western Tanzania.

Seasonal abundance, reproductive biology and feeding ecology of the bushveld gerbil Tatera leucogaster (Peters, 1852) were investigated in small-scale maize field-fallow land mosaics in south-western Tanzania. The gerbils were collected over a 2-year period using Sherman live and Victor hold-fast snap traps in permanent 4.5-ha grids. A total of 664 individuals were captured over 13 650 trap nights, giving an overall trap success rate of 4.9%. Trap success varied between seasons with and without crops in the field but not between habitat types. At this site, the breeding activity of this species is seasonal. All individuals whose stomachs were analyzed ate a wide range of items, indicating omnivory in this species at this site; however, seeds were the most preferred diet category, with a mean contribution of 50.4%, followed by arthropods, with a mean contribution of 25.7%. Other plant materials became important during the very dry periods.

Hair as an indicator of endogenous tissue levels of brominated flame retardants in mammals.

Few data are available on brominated flame retardants (BFRs) in terrestrial mammalian wildlife. Moreover, the use of hair in nondestructive monitoring of BFRs in mammals or humans has not been investigated. In the present study, concentrations of polybrominated diphenyl ethers (PBDEs) and brominated biphenyl 153 (BB 153) were analyzed in tissues of the European hedgehog Erinaceus europaeus. Road kills and carcasses from wildlife rescue centers were used to investigate relationships between concentrations of BFRs in hair and internal tissues, BFR tissue distribution (hair, liver, kidney, muscle, and adipose tissue), and PBDE congener tissue pattern dissimilarities. Liver concentrations of PBDEs and BB 153 were in the ranges 1-1178 and 0-2.5 ng/g of liver wet weight, respectively. PBDEs were predominant in adipose tissue and liver, while accumulation of BB 153 was tissue independent. The less persistent compound BDE 99 was more dominant in hair than in internal tissues. We observed positive relationships between BFR levels in hair and internal tissues for sum PBDEs and BDE 47 (0.37 < r < 0.78). The present study demonstrated that hair is a suitable indicator of PBDE exposure in terrestrial mammals which can be used in nondestructive monitoring schemes.

Host nutritive quality and host plant choice in two grass miners: primary roles for primary compounds?

The relationship between host plant choice and plant nutritional quality was investigated in two oligophagous grass miners Chromatomyia milii and C. nigra (Diptera, Agromyzidae). We tested whether host choice is determined by chemically mediated host suitability for offspring performance and/or adult performance. A second goal was to relate the observed variation among the different fitness parameters to quantitative and qualitative variation in foliar food quality. Choice experiments illustrated that both miners discriminated among grass species, and that C. milii has a smaller host range than C. nigra, as observed under natural conditions. Oviposition preference was correlated with adult feeding preference and related adult performance (longevity and fecundity) for both miners. Offspring performance measures (survival and pupal size) of at least C. nigra were more weakly related to host preference. Nearly all variation in adult performance of both miners was explained by foliar protein content, which had a positive effect on adult longevity and fecundity. Pupal size of both miners was positively related to foliar water and amino acid content and negatively related to lignin content. No clear relationship between host chemistry and offspring survival was observed. These observations show that fitness parameters are differentially related to host chemistry. Secondly, they suggest that chemically mediated host suitability for adult performance is an important determinant of host choice in this species. Finally, the results suggest a primary role for foliar protein content in host choice of the study species in general and in shaping the host range of C. milii in particular.