Femoral Structure and Biomechanical Characteristics in Sanfilippo Syndrome Type-B Mice.

Sanfilippo syndrome Type-B, also known as mucopolysaccharidosis IIIB (MPS IIIB), accounts for approximately one-third of all Sanfilippo syndrome patients and is characterized by a similar natural history as Type-A. Patients suffer from developmental regression, bone malformation, organomegaly, GI distress, and profound neurological deficits. Despite human trials of enzyme replacement therapy (ERT) (SBC-103, AX250) in MPS IIIB, there is currently no FDA approved treatment and a few palliative options. The major concerns of ERT and gene therapy for the treatment of bone malformation are the inadequate biodistribution of the missing enzyme, N-acetyl-α-glucosaminidase (NAGLU), and that the skeleton is a poorly hit target tissue in ERT and gene therapy. Each of the four known human types of MPS III (A, B, C, and D) is usually regarded as having mild bone manifestations, yet it remains poorly characterized. This study aimed to determine bone mineral content (BMC), volumetric bone mineral density (vBMD), and biomechanical properties in femurs MPS IIIB C57BL/6 mice compared to phenotypic control C57BL/6 mice. Significant differences were observed in MPS IIIB mice within various cortical and cancellous bone parameters for both males and females (p < 0.05). Here, we establish some osteogenic manifestations of MPS IIIB within the mouse model by radiographic and biomechanical tests, which are also differentially affected by age and sex. This suggests that some skeletal features of the MPS IIIB mouse model may be used as biomarkers of peripheral disease correction for preclinical treatment of MPS IIIB.

Antioxidant supplementation slows telomere shortening in free-living white stork chicks.

Telomere length (TL) and shortening is increasingly shown to predict variation in survival and lifespan, raising the question of what causes variation in these traits. Oxidative stress is well known to accelerate telomere attrition in vitro, but its importance in vivo is largely hypothetical. We tested this hypothesis experimentally by supplementing white stork (Ciconia ciconia) chicks with antioxidants. Individuals received either a control treatment, or a supply of tocopherol (vitamin E) and selenium, which both have antioxidant properties. The antioxidant treatment increased the concentration of tocopherol for up to two weeks after treatment but did not affect growth. Using the telomere restriction fragment technique, we evaluated erythrocyte TL and its dynamics. Telomeres shortened significantly over the 21 days between the baseline and final sample, independent of sex, mass, size and hatching order. The antioxidant treatment significantly mitigated shortening rate of average TL (-31% in shorter telomeres; percentiles 10th, 20th and 30th). Thus, our results support the hypothesis that oxidative stress shortens telomeres in vivo.

Antibiotic Perturbation of Gut Microbiota Dysregulates Osteoimmune Cross Talk in Postpubertal Skeletal Development.

Commensal gut microbiota-host immune responses are experimentally delineated via gnotobiotic animal models or alternatively by antibiotic perturbation of gut microbiota. Osteoimmunology investigations in germ-free mice, revealing that gut microbiota immunomodulatory actions critically regulate physiologic skeletal development, highlight that antibiotic perturbation of gut microbiota may dysregulate normal osteoimmunological processes. We investigated the impact of antibiotic disruption of gut microbiota on osteoimmune response effects in postpubertal skeletal development. Sex-matched C57BL/6T mice were administered broad-spectrum antibiotics or vehicle-control from the age of 6 to 12 weeks. Antibiotic alterations in gut bacterial composition and skeletal morphology were sex dependent. Antibiotics did not influence osteoblastogenesis or endochondral bone formation, but notably enhanced osteoclastogenesis. Unchanged Tnf or Ccl3 expression in marrow and elevated tumor necrosis factor-α and chemokine (C-C motif) ligand 3 in serum indicated that the pro-osteoclastic effects of the antibiotics are driven by increased systemic inflammation. Antibiotic-induced broad changes in adaptive and innate immune cells in mesenteric lymph nodes and spleen demonstrated that the perturbation of gut microbiota drives a state of dysbiotic hyperimmune response at secondary lymphoid tissues draining local gut and systemic circulation. Antibiotics up-regulated the myeloid-derived suppressor cells, immature myeloid progenitor cells known for immunosuppressive properties in pathophysiologic inflammatory conditions. Myeloid-derived suppressor cell-mediated immunosuppression can be antigen specific. Therefore, antibiotic-induced broad suppression of major histocompatibility complex class II antigen presentation genes in bone marrow discerns that antibiotic perturbation of gut microbiota dysregulates critical osteoimmune cross talk.

Urban blackbirds have shorter telomeres.

Urbanization, one of the most extreme human-induced environmental changes, represents a major challenge for many organisms. Anthropogenic habitats can have opposing effects on different fitness components, for example, by decreasing starvation risk but also health status. Assessment of the net fitness effect of anthropogenic habitats is therefore difficult. Telomere length is associated with phenotypic quality and mortality rate in many species, and the rate of telomere shortening is considered an integrative measure of the 'life stress' experienced by an individual. This makes telomere length a promising candidate for examining the effects of urbanization on the health status of individuals. We investigated whether telomere length differed between urban and forest-dwelling common blackbirds (Turdus merula). Using the terminal restriction fragment assay, we analysed telomere length in yearlings and older adults from five population dyads (urban versus forest) across Europe. In both age classes, urban blackbirds had significantly shorter telomeres (547 bp) than blackbirds in natural habitats, indicating lower health status in urban blackbirds. We propose several potential hypotheses to explain our results. Our findings show that even successful city dwellers such as blackbirds pay a price for living in these anthropogenic habitats.

Urbanization correlates with the prevalence and richness of blood parasites in Eurasian Blackbirds (Turdus merula).

Urbanization is increasing worldwide, producing severe environmental impacts. Biodiversity is affected by the expansion of cities, with many species being unable to cope with the different human-induced stressors present in these landscapes. However, this knowledge is mainly based on research from taxa such as plants or vertebrates, while other organisms like protozoa have been less studied in this context. The impact of urbanization on the transmission of vector-borne pathogens in wildlife is still unclear despite its relevance for animal and human health. Here, we investigated whether cities are associated with changes in the prevalence and richness of lineages of three vector-borne protozoans (Plasmodium, Haemoproteus and Leucocytozoon) in Eurasian blackbirds (Turdus merula) from multiple urban and forest areas in Europe. Our results show important species-specific differences between these two habitat types. We found a significant lower prevalence of Leucocytozoon in urban birds compared to forest birds, but no differences for Plasmodium and Haemoproteus. Furthermore, the richness of parasite lineages in European cities was higher for Plasmodium but lower for Leucocytozoon than in forests. We also found one Plasmodium lineage exclusively from cities while another of Leucocytozoon was only found in forests suggesting a certain level of habitat specialization for these protozoan vectors. Overall, our findings show that cities provide contrasting opportunities for the transmission of different vector-borne pathogens and generate new scenarios for the interactions between hosts, vectors and parasites.

Skeletal Effects of a Prolonged Oral Metformin Treatment in Adult Wistar Rats.

INTRODUCTION: We previously showed that a 3-week oral metformin (MET) treatment enhances the osteogenic potential of bone marrow stromal cells (BMSCs) and improves several bone histomorphometric parameters in Wistar rats with metabolic syndrome (MetS). However, the skeletal effects of extended periods of MET need to be completely elucidated. Hence, in this study, the impact of a prolonged (3-month) MET treatment was investigated on bone architecture, histomorphometric and biomechanics variables, and osteogenic potential of BMSCs in Wistar rats with or without MetS. MATERIALS AND METHODS: Young male Wistar rats (n=36) were randomized into four groups (n=9) that received either 20% fructose (F), MET (MET), F plus MET treatments (FMET), or drinking water alone (Veh). Rats were euthanized, blood was collected, and bones were dissected and processed for peripheral quantitative computed tomography (pQCT) analysis, static and dynamic histomorphometry, and bone biomechanics. In addition, BMSCs were isolated to determine their osteogenic potential. RESULTS: MET affected trabecular and cortical bone, altering bone architecture and biomechanics. Furthermore, MET increased the pro-resorptive profile of BMSCs. In addition, fructose-induced MetS practically did not affect the the structural or mechanical variables of the skeleton. CONCLUSION: A 3-month treatment with MET (with or without MetS) affects bone architecture and biomechanical variables in Wistar rats.

Influence of climatic conditions on the link between oxidative stress balance and landfill utilisation as a food resource by white storks.

Landfills have played a significant role in the recovery of white storks (Ciconia ciconia) populations across various European countries. While there is ample information about the populational-level effects, there is a lack of knowledge regarding the individual effects of using this food resource for feeding nestlings. This study aims to assess the nutritional status and oxidative stress balance of nestlings with varying degrees of exposure to landfill-provided food This study aims to assess the nutritional status and oxidative stress balance of nestlings with different use of landfill-provided food. Nestlings fed with food foraged by breeding pairs from landfills exhibited better nutritional status compared to individuals located farther from landfills. This can be attributed to a higher ingestion rate, resulting in increased plasmatic values of cholesterol, triglycerides, and HDL in plasma. However, the oxidative stress balance varied across different years, with individuals raised in 2014 showing higher values of Vitamin E and lower values of LDH compared to those raised in 2013. Furthermore, the impact of landfills on certain oxidative stress parameters also depended on the year of study. In 2013, the Total Antioxidant Capacity (TAC) of plasma showed a positive correlation with the distance to landfills, while the concentration of Malondialdehyde (MDA), an indicator of lipid peroxidation, exhibited a negative correlation. These findings suggest that the use of landfills as a food resource has a consistently positive effect on the nutritional status of white stork nestling. However, the relationship with oxidative stress is highly dependent on the climatic conditions of each year, emphasizing the importance of considering these factors when evaluating the use of landfills as a food resource.

The prevalence of anthropogenic nest materials differs between two distinct populations of migratory birds in Europe.

A major impact of human development is the transformation of natural habitats into farming lands and the expansion of built-up areas. Also, plastic pollution is affecting wildlife on a global scale. Discarded plastic is ubiquitous and accessible for birds, which can incorporate them into the nest structure. Here, we describe the differences in type, prevalence, and the amount of anthropogenic nest materials between two populations of terrestrial, mainly farmland bird, the white stork Ciconia ciconia, on a broad geographical scale, from two migratory divides-eastern in Poland and western in Spain (in total 303 nests). In the two populations, we detected significant differences in the incorporation of anthropogenic nest material, as measured by the Human Footprint Index (HFI) and the Impervious Surface Areas (ISA). We found that ISA was positively related to anthropogenic nest material incorporation in the Spanish population, and HFI was positively related to anthropogenic nest material, in contrast to the Polish population, in which the relationships were not significant. Moreover, we showed that the prevalence of nests with anthropogenic nest material was two times higher in Spanish than in the Polish white stork population. This study demonstrates that the behavior of incorporation of the anthropogenic nest material differs between two distinct populations of a single bird species.

Surveillance for gastrointestinal, subcutaneous, and ectoparasites of invasive North American raccoons (Procyon lotor) in central Spain.

The American raccoon (Procyon lotor) is an invasive meso-carnivore which has been introduced and established in many European countries. Although the presence of the raccoon in the Iberian Peninsula was confirmed around 20 years ago, there are few data on pathogens of these animals in this region. For this work, 72 American raccoons from two subpopulations in the central region of the Iberian Peninsula were examined for selected parasites. Ectoparasite species richness (both fleas and ticks) increased during the sampling season and was highest in the Henares subpopulation and on males. Similarly, ectoparasite abundance increased during the sampling season and was highest in Henares and on adult raccoons. Four species of ticks were detected including Rhipicephalus pusillus (71%), followed by R. sanguineus sensu lato (24%), Ixodes ventalloi (3%), and Dermacentor marginatus (1.4%). Four species of fleas were detected including Pulex irritans (44%), Ctenocephalides felis (3%), C. canis (1.4%), and Paraceras melis (1.4%) infestations. A subset of raccoons (n = 56) was examined for intestinal parasites; low prevalence and diversity were found including Strongyloides procyonis (4%), Dilepis sp. (5%), Plagiorchis sp. (2%), and Moniliformis moniliformis (2%). Importantly, Baylisascaris procyonis was not found. Finally, no subcutaneous nematodes (i.e., Dracunculus and Dirofilaria spp.) were found in the 56 raccoons examined. The results of this work show that the invasive North American raccoons currently are infected with few endoparasites but are commonly infested with native ectoparasites, several of which can transmit pathogens relevant for public and veterinary health. However, the geographically distinct populations of raccoons in Spain have different introduction histories, thus additional surveillance for parasites is warranted.

Environmental conditions but not nest composition affect reproductive success in an urban bird.

Adjusting the composition of their nests, breeding birds can influence the environmental conditions that eggs and offspring experience. Birds often use feathers to build nests, presumably due to their insulating properties. The amount of feathers in nests is often associated with increased nestling survival and body condition. However, it is unclear whether these putative beneficial effects of adding feathers to nests are relevant in a wide range of environmental conditions. Here, we combine data on weather conditions and feathers in nests (i.e., nest composition) to investigate their relative contribution to reproductive success in the Eurasian tree sparrow (Passer montanus). Specifically, we investigate whether the effect of weather conditions on breeding success is modulated by the amount of feathers added to the nest. We found a strong negative effect of rainfall on the number of nestlings that successfully fledged per breeding attempt, but this negative effect was not mitigated by the amount of feathers in nests. We also found that the amount of feathers in nests varied along the breeding season, with nests containing more feathers early in the breeding season, when temperatures were lower. Despite considerable variation in nest composition, our results do not suggest an important role of feathers in nests protecting eggs or nestling tree sparrows against fluctuations in environmental conditions.

The trade-offs of foraging at landfills: Landfill use enhances hatching success but decrease the juvenile survival of their offspring on white storks (Ciconia ciconia).

During the last decades, landfills have become a valuable food source for wildlife, being in some cases determinants of large avian population increases. Superabundant food resources at landfills can increase reproductive and/or survival parameters; however, negative effects such as intoxication, plastic ingestion, skeletal deformities, unbalanced oxidative stress, and other health problems have also been reported. White stork (Ciconia ciconia) commonly benefits from landfill resources. Here, we evaluate potential landfill effects on demographic parameters (reproduction and offspring survival) at the individual level in a single population. Our results show that a more intense use of landfills by breeders has a positive effect on hatching success but a negative effect on juvenile survival probability after emancipation, at least during the first year of life. High amount of food and proximity to landfill may explain their beneficial effect on reproductive parameters. On the other hand, poor quality food, pollutants, and pathogens acquired during early development from a diet based on refuse may be responsible for reduced future survival probability. Consequently, both positive and negative effects were detected, being foraging at landfills at low to medium levels the better strategy. Although our study shows that intense foraging on rubbish can imply both costs and benefits at an individual level, the benefits of superabundant food provisioning observed at population level by other studies cannot be ignored. Management actions should be designed to improve natural food resources, reduce non-natural mortality and/or human disturbances to guarantee the species viability under current European Union regulations designed to ban open-air landfills in a near future.

A multidisciplinary approach to the evaluation of the effects of foraging on landfills on white stork nestlings.

The use of landfills as foraging areas by white storks (Ciconia ciconia) is a recent well-known behaviour. While several studies have highlighted positive effects at a populational level others suggest that the presence of pollutants, pathogens and the lower presence of antioxidants in the food could pose a health risk for individuals. The objective of this study was to evaluate potential effects of the use of landfills as a food resource on the physiology and health of white stork nestlings, by a multidisciplinary approach based on the analysis of nutritional status, body condition, blood parameters, oxidative stress balance and the presence of pathogens. Results showed better body condition in individuals associated with landfills compared to the ones feeding on natural resources, as well as better nutritional status, as indicated by higher levels of albumin, cholesterol, and triglycerides in plasma. As many pollutants have a pro-oxidant effect, we evaluated oxidative stress balance, with no differences in the indicators of damage except for methaemoglobin (metHb), significantly higher in nestlings associated with landfill-origin food. Regarding antioxidants, GSH was higher in nestlings associated with landfills, which may suggest a hormetic response induced potentially by the presence of pollutants in waste. Nestlings fed food from landfills also had a higher presence of Escherichia coli with a multiresistant phenotype to antibiotics. In conclusion, our results show that nestlings fed with a higher proportion of food from landfills present a better nutritional status and body condition than those fed with a higher proportion of natural diet, being the only indicators of negative effects of the use of this food resource the higher percentage of metHb in the peripheral blood and the presence of antibiotic-resistant E. coli.

Distance to landfill and human activities affects the debris incorporation into the white stork nests in urbanized landscape in central Spain.

Human's activities dominates many aspects of the Earth's environment; thus animals are forced to adapt and respond to the resulting changes in habitat structure and functioning due to anthropogenic pressure. Along with the growing human population and the associated amount of waste produced, the amount of different type of physical contamination component in environment is increasing. Incorporation of debris in nests may be a mounting avian response to anthropogenic pollution. In this research, we quantified the constituent pieces and total mass of human-derived materials incorporated in white stork nests. The study was conducted on four locations in central Spain where white storks nest along a urbanization gradient. In total, we examined 49 nests. This study demonstrates that the incorporation of debris by white storks into their nests is related to human activity, measured by the Human Footprint Index (HFI). Moreover, the distance between these nests and landfills predicts the occurrence of debris incorporated into nests. Our study shows that birds nest building behaviour is impacted by human activities and pollution in environment.

Physiological stress does not increase with urbanization in European blackbirds: Evidence from hormonal, immunological and cellular indicators.

Urbanization changes the landscape structure and ecological processes of natural habitats. While urban areas expose animal communities to novel challenges, they may also provide more stable environments in which environmental fluctuations are buffered. Species´ ecology and physiology may determine their capacity to cope with the city life. However, the physiological mechanisms underlying organismal responses to urbanization, and whether different physiological systems are equally affected by urban environments remain poorly understood. This severely limits our capacity to predict the impact of anthropogenic habitats on wild populations. In this study, we measured indicators of physiological stress at the endocrine, immune and cellular level (feather corticosterone levels, heterophil to lymphocyte ratio, and heat-shock proteins) in urban and non-urban European blackbirds (Turdus merula) across 10 European populations. Among the three variables, we found consistent differences in feather corticosterone, which was higher in non-urban habitats. This effect seems to be dependent on sex, being greater in males. In contrast, we found no significant differences between urban and non-urban habitats in the two other physiological indicators. The discrepancy between these different measurements of physiological stress highlights the importance of including multiple physiological variables to understand the impact of urbanization on species' physiology. Overall, our findings suggest that adult European blackbirds living in urban and non-urban habitats do not differ in terms of physiological stress at an organismal level. Furthermore, we found large differences among populations on the strength and direction of the urbanization effect, which illustrates the relevance of spatial replication when investigating urban-induced physiological responses.

Life in a polluted world: A global review of anthropogenic materials in bird nests.

Human pressure exerts a significant influence on animals and the environment. One of its consequences, plastic pollution is considered one of the major threats to fauna as well as a significant conservation issue. In this research, we examined the global pattern of one example of avian behavior in response to pollution-namely, the incorporation of anthropogenic materials into nests-as well as the existing knowledge on this subject. Based on 25 articles, we studied 51 populations, involving 24 bird species, and checked 10,790 nests; as a result, we found that incorporation of debris is correlated with increasing human influence on the environment, measured as the Human Footprint Index. Moreover, the probability of debris incorporation is higher in terrestrial than in marine species. We also identified knowledge bias in favor of marine as opposed to terrestrial species: namely, marine species attract more scientific attention than terrestrial. Furthermore, research approaches to these two ecosystems differ. Undeniably, the factors which influence debris incorporation by birds, the scale of this behavior, and particular forms of use of debris in bird nests are aspects which require long-term standardized research. This is the first global review paper on debris incorporation by birds to demonstrate a close link to human pressure as a driver.

Influence of trophic ecology on the accumulation of dioxins and furans (PCDD/Fs), non-ortho polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) in Mediterranean gulls (Larus michahellis and L. audouinii): A three-isotope approach.

The impact of pollution caused by severe anthropogenic pressure in the Mediterranean Sea, an important biodiversity hotspot, requires continuous research efforts. Sources of highly toxic chemicals such as Persistent Organic Pollutants (POPs) are misunderstood in representative Mediterranean species, which limits our capability to establish proper conservation strategies. In the present study, eggs of Audouin's and yellow-legged gulls (Larus audouinii and L. michahellis) were used to investigate the trophic sources, as measured by δ(13)C, δ(15)N, and δ(34)S, of legacy POPs, in particular, polychlorinated dibenzo-p-dioxins and furans (PCDD/Fs) and non-ortho polychlorinated biphenyls (no-PCBs), as well as recently-regulated POPs, e.g., polybrominated diphenyl ethers (PBDEs). Special attention was paid to the usefulness of rarely-explored δ(34)S ratios in explaining POP exposure in wildlife, and δ(34)S was the isotopic ratio that best explained POP variations among gulls in most cases, thus demonstrating its usefulness for understanding POP exposure in wildlife. Significant relationships between stable isotope signatures and POP concentrations revealed increasing levels of no-PCBs and low halogenated PCDD/Fs and PBDEs in Mediterranean gulls as the consumption of marine resources increases. In contrast, highly chlorinated and brominated congeners appeared to preferentially accumulate in gulls feeding primarily on refuse from dump sites and terrestrial food webs. The use of suitable dietary tracers in the study of POPs in yellow-legged gulls revealed the importance of dump sites as a source of POPs in Mediterranean seabirds, which has not previously been reported. In contrast, the preferential accumulation through marine food webs of low chlorinated PCCD/Fs and no-PCBs, which show the highest toxic equivalents factors (TEFs), led to a significantly greater toxicological concern in Audouin's as compared to yellow-legged gulls. Audouin's gull exposure to POPs appears primarily related to the pelagic food webs commonly exploited by fisheries, highlighting the need for further research given the potential impact on human consumption.

Predominance of BDE-209 and other higher brominated diphenyl ethers in eggs of white stork (Ciconia ciconia) colonies from Spain.

Polybrominated diphenyl ethers (PBDEs) are ubiquitous pollutants for which there is still a lack of knowledge about the environmental behavior and fate of the higher brominated congeners (octa- to deca-BDEs). In this study, the PBDE content and congener profiles in failed eggs from two colonies of white stork (Ciconia ciconia) in Spain were studied. The average total PBDE concentration was 1.64ng/g (wet weight, w.w.) for the rural colony and 9.08ng/g (w.w.) for the urban colony. Higher brominated BDEs dominated the congener profiles of both colonies. Of particular interest was the determination of BDE-209 as the dominant congener accounting for 44.1% and 38.6% of the total PBDE content in the rural and urban colonies, respectively. BDE-202, considered an indicator of BDE 209 debromination, was detected in 83% and all of the samples from rural and urban colonies, respectively. The observed congener profile in which BDE-207>BDE-208>BDE-206 does not correspond to any known technical PBDE mixture and is evidence for possible BDE-209 degradation.