Long-term Exposure to Industrial Chemical Contamination Affects the Magnitude of Predator-induced Immunosuppression in a Free-living Passerine.

Industrial chemical contamination is known to have immuno-toxic effects on birds. It may also interfere with natural stressful conditions to further disrupt the immune responses, but these possible interactive effects are still poorly documented in free-living birds. Using the phytohaemagglutinin skin-swelling test, we assessed how the T-cell mediated immune response varied according to the perceived risk of predation in hybrid sparrows, Passer domesticus × Passer hispaniolensis, originating from two sites differentially impacted by industrial chemical contamination, in southern Tunisia. Results showed that T-cell mediated immune response decreased with increasing perceived risk of predation, but the extent of this predator-associated immunosuppression was weaker in birds from the contaminated site compared to those from the control site. The immune response of birds living in the contaminated site was so weak that it could not be further weakened by a predator-related stress. Overall, these results support the idea that chemical contamination interferes with natural environmental stressors, such as predators, thus entailing profound disruption of the immune responses, with possible deleterious repercussions on the ability of birds to cope with diseases.

Experimental evidence of increased carcass removal along roads by facultative scavengers.

Investigating the spatial response of scavenging behaviors to roads may help in understanding the relevance of this overlooked ecosystem service. Roads can provide suitable foraging sites for scavengers, whether they are obligate or facultative. However, only a few studies have investigated the impact of roads on the spatial distribution of scavenging behavior, and most of them focused on areas inhabited by species-rich communities of obligate scavengers. In this study, we investigated this issue in a poorly productive arid environment in southern Tunisia where the main facultative scavengers were stray dogs. Our experimental design was based on the monitoring of carcasses placed at different distances from the road. We found that carcasses were removed faster along roadsides than farther away, showing the important role of road traffic in shaping the spatial distribution of scavenging activity. These results differ from those found by similar experimental studies also conducted on facultative scavengers but in more productive environments. Indeed, given the scarcity of food resources in the arid study area, scavengers seemed to depend largely on road-kills for food, thus concentrating their foraging activity along the roads in accordance with the predictions of the optimal foraging theory. Overall, our findings represent clear evidence that roads and related traffic can alter the spatial distribution of ecosystem services, with potential implications for food webs and ecosystem functioning. They also stress the need for accounting for scavengers when performing road-kill surveys, but also considering road effects when carrying out monitoring of the activity of scavengers.

Behavioral fever in Bosk's fringe-toed lizards (Acanthodactylus boskianus) living in an industrial area in south-eastern Tunisia.

As an indicator of physiological state of lizards, thermoregulatory behavior has been proposed as an assessment tool of environmental contamination, particularly in desert ecosystems where lizards represent a major biological component. Although pesticide contamination has been shown to induce behavioral fever in lizards, the possible pyrogenic effect of heavy metals has received less attention. The objective of this study was to provide data on this issue by using Bosk's fringe-toed lizards (Acanthodactylus boskianus) living in a metal-contaminated industrial area in south-eastern Tunisia as a study model. Combining field observations of behavior with measures of thermoregulatory performance under laboratory conditions, we found that proximity to the industrial zone was associated to a different thermoregulatory behavior of lizards. There was a tendency of lizards living close to the contamination source to display behavioral fever, as they spent more time in the sun, preferred higher body temperatures, and warmed up faster, compared to lizards living far away. This heat-seeking behavior is likely a reaction to improve the functioning of physiological systems involved in the defense against toxic effects of heavy metals, possibly through the increase of metabolic rate, stimulation of enzymes involved in the detoxification process, and enhancement of immune defense systems. Our findings stress the need for further detailed studies into the physiological mechanisms linking metal contamination to behavioral fever. They also underline the usefulness of thermoregulatory behavior as a reliable, non-destructive, and inexpensive, individual-level biomarker in lizard ecotoxicology and biomonitoring of contaminated desert environments.

Prioritizing the provision of urban ecosystem services in deprived areas, a question of environmental justice.

The distribution of urban ecosystem services (UES) is often uneven across socioeconomic groups, leading to environmental justice issues. Understanding the distribution of UES across a landscape can help managers ensure an equitable distribution of services. While many past studies have focused on the distribution of green spaces in relation to socioeconomic variables, this research analyzes the distribution of UES provided by these green spaces. This research quantified air pollution removal, atmospheric carbon reduction, and surface runoff mitigation provided by urban trees in Strasbourg city (France). The provision of these three UES was studied at the census block scale by creating an index of UES delivery, which was contrasted with a constructed social deprivation index. Our results show that there is no significant association between the delivery of UES and social deprivation. Some deprived populations benefit from high UES delivery. Results also suggest that mapping associations between UES delivery and social deprivation should be integrated with future development plans to enhance the equitable distribution of UES. This study provides insights into the French context where studies about the distribution of UES at a small-area level remain lacking.

Decreased Cell-Mediated Immune Response in Bosk's Fringe-Toed Lizards (Acanthodactylus boskianus) Inhabiting an Industrialized Area in Southern Tunisia.

Lizards increasingly are recognized as suitable contaminant biomonitors in terrestrial ecosystems. Previously, we have shown that Bosk's fringe-toed lizards (Acanthodactylus boskianus) living close to the Gabès-Ghannouche industrial complex for fertilizer and acid production in southern Tunisia were contaminated by heavy metals. However, the impact of this contamination on lizard health parameters has not been investigated. In this study, we used the phytohaemagglutinin (PHA) skin-swelling test to assess whether the proximity to the industrial complex was associated with notable changes in lizard cell-mediated immune response (CMI). Our results showed significantly lower CMI in lizards living close to the industrial complex compared to those occurring farther away in a similar coastal habitat. Overall, our findings are consistent with the idea of immunotoxic effects of metal contamination. They also stress the usefulness of the PHA approach as an efficient tool for the evaluation of contaminant-related immunosuppression in lizards.

Flaviviruses in migratory passerines during spring stopover in a desert oasis.

Bird migration has long been hypothesized as the main mechanism for long-distance dispersal of flaviviruses, but the role of migratory birds in flaviviruses spillover is not well documented. In this study, we investigated the eco-epidemiology of West Nile virus (WNV) and Usutu virus (USUV) in trans-Saharan passerines during their spring stopover in southern Tunisian oases. To do, we combined oral swab analysis and serological tools to assess whether migratory birds could be reaching these stopover sites while infectious or have been previously exposed to viruses. All sampled birds tested negative for oral swab analysis. However, anti-WNV and anti-USUV antibodies were detected in 32% and 1% of tested birds, respectively. Among WNV-seropositive species, the Golden oriole (Oriolus oriolus) showed the highest anti-WNV occurrence probability. In this species, anti-WNV occurrence was twice larger in males than females. Inter-specific and intraspecific morphological, physiological and behavioural differences could explain these results. Although our findings did not show evidence for passerines migrating while infectious, they did not exclude an existing enzootic WNV transmission cycle in Tunisian oases. Further investigations including larger samples of migratory birds are needed for a better understanding of this issue.

Exposure of yellow-legged gulls to Toxoplasma gondii along the Western Mediterranean coasts: Tales from a sentinel.

Efficiently tracking and anticipating the dynamics of infectious agents in wild populations requires the gathering of large numbers of samples, if possible at several locations and points in time, which can be a challenge for some species. Testing for the presence of specific maternal antibodies in egg yolks sampled on the colonies could represent an efficient way to quantify the exposure of breeding females to infectious agents, particularly when using an abundant and widespread species, such as the yellow-legged gull (Larus michahellis). We used such an approach to explore spatio-temporal patterns of exposure to Toxoplasma gondii, a pathogenic protozoan responsible of toxoplasmosis in humans and other warm-blooded vertebrates. First, we tested the validity of this approach by exploring the repeatability of the detection of specific antibodies at the egg level using two different immunoassays and at the clutch level using an occupancy model. Then, samples gathered in 15 colonies from France, Spain and Tunisia were analysed using an immunoassay detecting antibodies specifically directed against T. gondii. Prevalence of specific antibodies in eggs was overall high while varying significantly among colonies. These results revealed that T. gondii circulated at a large spatial scale in the western Mediterranean yellow-legged gull population, highlighting its potential role in the maintenance community of this parasite. Additionally, this study illustrates how species commensal to human populations like large gulls can be used as wildlife sentinels for the tracking of infectious agents at the human-wildlife interface, notably by sampling eggs.

Diversity, prevalence and host specificity of avian parasites in southern Tunisian oases.

Oases are semi-natural woodlots surrounded by an inhospitable desert environment. This insular-like habitat system is known to support a mixture of sedentary and migratory bird species originating from different areas. However, little is known about the interactions between these birds and parasites. In this study, we investigated the diversity, prevalence and host specificity of avian haemosporidian parasites in southern Tunisian oases in two sedentary and common bird species, the laughing dove Spilopelia senegalensis and hybrid sparrow Passer domesticus × hispaniolensis, in six sites that differ regarding vegetation structure and distance to the coast. Two new Haemoproteus lineages, related to other Haemoproteus transmitted by biting midges, were detected in doves. With regard to sparrows, all detected parasites have previously been reported in other sparrow populations, except for one new Haemoproteus lineage. Our results also showed that densely vegetated sites were characterized by the higher prevalence of Plasmodium but a lower prevalence of Haemoproteus compared with less-vegetated sites. This is the first study aiming to explore avian parasites in the oasis habitat. Gathering data on a larger sample of oases with different sizes and isolation levels will be the next step to better understand factors shaping the transmission dynamics of avian parasites in such ecosystems.

Heavy metal accumulation in lizards living near a phosphate treatment plant: possible transfer of contaminants from aquatic to terrestrial food webs.

We investigated the accumulation of heavy metals in Bosk's fringe-toed lizards (Acanthodactylus boskianus) living in Gabès region (southeastern Tunisia), in relation to habitat, diet, and distance from the Gabès-Ghannouche factory complex of phosphate treatment. More specifically, we compared the concentrations of cadmium, lead, and zinc in the stomach contents and samples of the liver, kidney, and tail from lizards living in four sites corresponding to different combinations of habitat (coastal dunes vs backshore) and distance from the factory complex (<500 vs 20 km). Examination of stomach contents showed that lizards living on the coastal dunes mainly feed on littoral amphipods, while those living in the backshore feed exclusively on terrestrial invertebrates. The concentrations of heavy metals in lizard tissues were overall positively correlated with those in the preys they ingested. Moreover, there was a general tendency towards increased concentrations of cadmium, lead, and zinc in the samples from lizards living on coastal dunes compared to those from the other sites, although some differences still lacked statistical significance. These results suggest that the highest contamination of lizards living on coastal dunes was probably related to the ingestion of contaminated amphipods. Thus, amphipods and Bosk's fringe-toed lizards seem to provide an important link between the marine and terrestrial food webs, with higher concentrations appearing to accumulate from materials released into the sea rather than the terrestrial environment. With regard to metal distribution among tissues, our results were overall in agreement with previous findings in other reptiles. In particular, cadmium was most concentrated in the liver samples, stressing once more the role of the liver as a storage organ of Cd. Moreover, high concentrations of the three assessed metals were found in the kidney samples, showing the role of the kidney as an active site of heavy metal accumulation.

Reproductive effects in hybrid sparrow from a polluted area in Tunisia: Oxidative damage and altered testicular histomorphology.

Air pollution is a threat for human health and wildlife. The aim of this study is to assess the pathophysiological changes and the oxidative-antioxidative status in testicular tissues of 40 Hybrid sparrows collected from four areas in Gabès city, one of the most polluted areas in Tunisia. The testis histopathological analysis revealed alterations in birds from Ghannouche, the polluted area. The thiobarbituric acid reactive substance (TBARS) levels were higher in testis of birds from the contaminated site compared to less polluted areas indicating oxidative damage to membrane lipids. Antioxidant enzyme activities (superoxide dismutase and catalase) were lower in testis sparrows from the polluted site compared with the reference site, suggesting deficiency of the antioxidant system to compensate for oxidative stress. Overall, our results suggest that the hybrid sparrow offers a suitable model for biomonitoring programs of atmosphere pollutants and the selected biomarkers could be useful tool to evaluate pollution impacts in living organisms.

Condition and Health of Rufous Bush Robin (Cercotrichas galactotes) Nestlings in a Polluted Oasis Habitat in Southern Tunisia.

We investigated whether the proximity to the Gabès-Ghannouche factory complex of phosphate treatment, in south-eastern Tunisia, was associated with notable changes in the condition and health of Rufous bush robin (Cercotrichas galactotes) nestlings hatched in the neighbouring oasis habitat. Results demonstrated that excrements of nestlings hatched in one oasis close to the factory complex contained higher concentrations of lead and zinc than the excrements of nestlings from one oasis situated 20 km away. Furthermore, when effects of age and nestling number in the nest were controlled, nestlings hatched near the factory complex showed reduced tarsus length, rectrix length, haematocrit level and haemoglobin concentration, but higher levels of fluctuating asymmetry compared to those from the more distant oasis. Overall, results suggest that the proximity to the factory complex was associated with increased exposure to metals and deterioration in nestling condition and development.

Egg sampling as a possible alternative to blood sampling when monitoring the exposure of yellow-legged gulls (Larus michahellis) to avian influenza viruses.

We explored whether antibody detection in egg yolks could serve as an alternative to antibody detection in plasma samples when monitoring yellow-legged gulls (Larus michahellis) for exposure to avian influenza viruses (AIVs). We tested female plasma and eggs for anti-AIV antibodies and used the data we obtained to check whether the two sample types yielded the same antibody status (positive or negative) and to compare the antibody prevalence estimated from the blood data with that estimated from the yolk data. Our results showed that sampling one egg per clutch, regardless of that egg's position in the laying sequence, is sufficient to provide an unbiased estimate of antibody prevalence across clutches. The results also showed that almost 25% of the clutches laid by positive females contained only antibody-negative eggs, which suggests that yolk samples might underestimate female antibody prevalence. However, this result may stem from differences in the methods used to assess plasma versus yolk antibody status. Further research is needed to clarify this issue; while the number of false negatives could be reduced by adapting antibody detection techniques, it may be that they are an unavoidable consequence of natural avian maternal transfer dynamics.

Circulation of a Meaban-like virus in yellow-legged gulls and seabird ticks in the western Mediterranean basin.

In recent years, a number of zoonotic flaviviruses have emerged worldwide, and wild birds serve as their major reservoirs. Epidemiological surveys of bird populations at various geographical scales can clarify key aspects of the eco-epidemiology of these viruses. In this study, we aimed at exploring the presence of flaviviruses in the western Mediterranean by sampling breeding populations of the yellow-legged gull (Larus michahellis), a widely distributed, anthropophilic, and abundant seabird species. For 3 years, we sampled eggs from 19 breeding colonies in Spain, France, Algeria, and Tunisia. First, ELISAs were used to determine if the eggs contained antibodies against flaviviruses. Second, neutralization assays were used to identify the specific flaviviruses present. Finally, for colonies in which ELISA-positive eggs had been found, chick serum samples and potential vectors, culicid mosquitoes and soft ticks (Ornithodoros maritimus), were collected and analyzed using serology and PCR, respectively. The prevalence of flavivirus-specific antibodies in eggs was highly spatially heterogeneous. In northeastern Spain, on the Medes Islands and in the nearby village of L'Escala, 56% of eggs had antibodies against the flavivirus envelope protein, but were negative for neutralizing antibodies against three common flaviviruses: West Nile, Usutu, and tick-borne encephalitis viruses. Furthermore, little evidence of past flavivirus exposure was obtained for the other colonies. A subset of the Ornithodoros ticks from Medes screened for flaviviral RNA tested positive for a virus whose NS5 gene was 95% similar to that of Meaban virus, a flavivirus previously isolated from ticks of Larus argentatus in western France. All ELISA-positive samples subsequently tested positive for Meaban virus neutralizing antibodies. This study shows that gulls in the western Mediterranean Basin are exposed to a tick-borne Meaban-like virus, which underscores the need of exploring the spatial and temporal distribution of this flavivirus as well as its potential pathogenicity for animals and humans.

Maternal antibody transmission in relation to mother fluctuating asymmetry in a long-lived colonial seabird: the yellow-legged gull Larus michahellis.

Female birds transfer antibodies to their offspring via the egg yolk, thus possibly providing passive immunity against infectious diseases to which hatchlings may be exposed, thereby affecting their fitness. It is nonetheless unclear whether the amount of maternal antibodies transmitted into egg yolks varies with female quality and egg laying order. In this paper, we investigated the transfer of maternal antibodies against type A influenza viruses (anti-AIV antibodies) by a long-lived colonial seabird, the yellow-legged gull (Larus michahellis), in relation to fluctuating asymmetry in females, i.e. the random deviation from perfect symmetry in bilaterally symmetric morphological and anatomical traits. In particular, we tested whether females with greater asymmetry transmitted fewer antibodies to their eggs, and whether within-clutch variation in yolk antibodies varied according to the maternal level of fluctuating asymmetry. We found that asymmetric females were in worse physical condition, produced fewer antibodies, and transmitted lower amounts of antibodies to their eggs. We also found that, within a given clutch, yolk antibody level decreased with egg laying order, but this laying order effect was more pronounced in clutches laid by the more asymmetric females. Overall, our results support the hypothesis that maternal quality interacts with egg laying order in determining the amount of maternal antibodies transmitted to the yolks. They also highlight the usefulness of fluctuating asymmetry as a sensitive indicator of female quality and immunocompetence in birds.

Breeding performance of passerines in a polluted oasis habitat in southern Tunisia.

We investigated the breeding performance of passerines nesting in the oases close to Gabès city, one of the most industrialised areas in Tunisia, in relation to habitat pollution. Using data on nest survival, clutch size and chick productivity of four insectivorous passerine species, we assessed whether the proximity to Gabès-Ghannouche factory complex of phosphate treatment was associated with reduced breeding performance. Our results show evidence of decreased breeding performance near the factory complex, but this decrease was more perceptible when comparing one oasis situated at the vicinity of the factory complex (polluted oasis) with one control oasis situated tens of kilometres faraway, than when comparing sites situated hundreds of metres apart within the polluted oasis. Overall, these results would indicate that passerines offer suitable models for biomonitoring programmes in the Gabès region and underline once more the usefulness of passerines as reliable biomonitors of terrestrial ecosystems.

Prevalence of influenza A antibodies in yellow-legged gull (Larus michahellis) eggs and adults in southern Tunisia.

Investigating the prevalence of anti-influenza A viruses (AIV) antibodies in wild birds can provide important information for the understanding of bird exposure to AIV, as well as for prevention purposes. We investigated AIV exposure in nature by measuring the prevalence of anti-AIV antibodies in the nests and adults of an abundant and anthropophilic waterbird species common around the Mediterranean sea, the yellow-legged gull (Larus michahellis). Sampling took place in two colonies located in the gulf of Gabès in southern Tunisia: Sfax and Djerba. Antibodies were detected in the two sites, with higher prevalence in adults, eggs, and nests at Sfax than Djerba. Across both colonies, clutches that were laid later in the season, and, thus, more likely by younger parents, showed lower prevalence. Using patch occupancy modeling applied to egg clutches, we found that it is unnecessary to sample all the eggs in a given nest; nest status (antibody positive or negative) can be reliably estimated from a single egg. Differences in the density of birds, notably Larids, between the two sites may explain the observed differences in prevalence. The higher concentration of Larids in the Sfax colony could favor the transmission of AIV to yellow-legged gulls. This study highlights the importance of further developing ecological-based approaches to the factors determining the circulation of infectious agents in species such as the yellow-legged gull, which exist at the interface between diverse biological communities and human activities.

Distribution-abundance relationship for passerines breeding in Tunisian oases: test of the sampling hypothesis.

The positive relationship between local abundance and distribution of species is a widely recognized pattern in community ecology. However, it has been suggested that this relationship can simply be an artefact of sampling because locally rare species are less detectable then locally abundant ones, and hence their distribution can easily be underestimated. Here, we use count data to investigate the relationship between distribution and abundance of passerines breeding in a sample of oases from southern Tunisia, and we provide a test of the sampling artefact hypothesis. In particular, we checked for a difference in detection probability between localized and widespread species, and we tested if increasing the sampling effort affects the significance of the relationship. A significant positive relationship between the average local abundance of passerine species and the proportion of occupied oases was found. The use of a capture-recapture approach allowed us to estimate and to compare the detection probabilities of localized and widespread species subsets. We found that localized species were locally less detectable than widespread species, which is consistent with the main assumption of the sampling artefact hypothesis. However, increasing the detection probability of species by conducting more counts did not affect the significance of the relationship, which did not give support to the sampling artefact hypothesis. Our work implies that sampling contributed to the distribution-abundance relationship we found, but that it is unlikely that such a relationship could entirely be explained by an artefact of sampling. It also underlines the insight that can be gained by using probabilistic approaches of estimating species number and detection probability when attempting to disentangle sampling from ecological effects in community ecology studies.