Tick infestation in spur-thighed tortoise population: a pilot study for unraveling epidemiological patterns and demographic consequences.

Ectoparasites, such as ticks, modulate host population dynamics by impacting demographic traits. They transmit infectious agents among their hosts, posing a critical threat to animal and public health. This study aimed to characterize and analyze the Hyalomma aegyptium infestation on one of its main hosts, the spur-thighed tortoise, its effects on demographic traits, and to determine the diversity of infectious agents present in both ticks and tortoises in the Maamora forest (northwestern Morocco). Our results show that 100% of the tortoises were parasitized by adult ticks in spring, an infestation intensity of 4 ticks/tortoise (5.1 and 3.6 ticks/tortoise in males and females, respectively; 4.2 and 3.3 ticks/tortoise in gravid and non-gravid females, respectively) and an abundance ranging from 1 to 12. Although without significant differences, male tortoises had higher tick abundances than females. The interaction of tortoise sex and body condition was significantly related to tick abundance, male body condition decreased with higher tick abundance in contrast to females. Nevertheless, the interaction of body condition and reproductive stage of females was not significantly related to tick abundance. Gravid females were significantly associated with tick abundance, showing a slightly higher infestation than non-gravid females. Molecular analysis of pooled tick samples revealed the presence of Ehrlichia ewingii, Candidatus Midichloria mitochondrii, and Rickettsia africae, with a minimum infection rate of 0.61 to 1.84%. However, blood sample analysis of the tortoises was infectious agent-free, pinpointing a lack of significant health problems. Given the possible effect on the transmission of zoonotic diseases by spur-thighed tortoises associated with their frequent collection as pets, it should be surveyed to control possible human health problems. In conservation terms, as a long-lived species, the role of tick infestation in demographic traits might be included in the management and conservation programs of spur-thighed tortoises.

Exploiting the multiplexing capabilities of tandem mass tags for high-throughput estimation of cellular protein abundances by mass spectrometry.

The generation of dynamic models of biological processes critically depends on the determination of precise cellular concentrations of biomolecules. Measurements of system-wide absolute protein levels are particularly valuable information in systems biology. Recently, mass spectrometry based proteomics approaches have been developed to estimate protein concentrations on a proteome-wide scale. However, for very complex proteomes, fractionation steps are required, increasing samples number and instrument analysis time. As a result, the number of full proteomes that can be routinely analyzed is limited. Here we combined absolute quantification strategies with the multiplexing capabilities of isobaric tandem mass tags to determine cellular protein abundances in a high throughput and proteome-wide scale even for highly complex biological systems, such as a whole human cell line. We generated two independent data sets to demonstrate the power of the approach regarding sample throughput, dynamic range, quantitative precision and accuracy as well as proteome coverage in comparison to existing mass spectrometry based strategies.

Abiotic, biotic, and evolutionary control of the distribution of C and N isotopes in food webs.

Ecosystem functioning depends on nutrient cycles and their responses to abiotic and biotic determinants, with the influence of evolutionary legacies being generally overlooked in ecosystem ecology. Along a broad elevation gradient characterized by shifting climatic and grazing environments, we addressed clines of plant N and C∶N content and of δ(13)C and δ(15)N in producers (herbs) and in primary (grasshoppers) and secondary (birds) consumers, both within and between species in phylogenetically controlled scenarios. We found parallel and significant intra- and interspecific trends of isotopic variation with elevation in the three groups. In primary producers, nutrient and isotope distributions had a detectable phylogenetic signal that constrained their variation along the environmental gradient. The influence of the environment could not be ascribed to any single factor, and both grazing and climate had an effect on leaf stoichiometry and, thus, on the resources available to consumers. Trends in consumers matched those in plants but often became nonsignificant after controlling for isotopic values of their direct resources, revealing direct bottom-up control and little phylogenetic dependence. By integrating ecosystem and mechanistic perspectives, we found that nutrient dynamics in food webs are governed at the base by the complex interaction between local determinants and evolutionary factors.

Synergistic Effects of PARP Inhibition and Cholesterol Biosynthesis Pathway Modulation.

An in-depth multiomic molecular characterization of PARP inhibitors revealed a distinct poly-pharmacology of niraparib (Zejula) mediated by its interaction with lanosterol synthase (LSS), which is not observed with other PARP inhibitors. Niraparib, in a similar way to the LSS inhibitor Ro-48-8071, induced activation of the 24,25-epoxysterol shunt pathway, which is a regulatory signaling branch of the cholesterol biosynthesis pathway. Interestingly, the combination of an LSS inhibitor with a PARP inhibitor that does not bind to LSS, such as olaparib, had an additive effect on killing cancer cells to levels comparable with niraparib as a single agent. In addition, the combination of PARP inhibitors and statins, inhibitors of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, an enzyme catalyzing the rate-limiting step in the mevalonate pathway, had a synergistic effect on tumor cell killing in cell lines and patient-derived ovarian tumor organoids. These observations suggest that concomitant inhibition of the cholesterol biosynthesis pathway and PARP activity might result in stronger efficacy of these inhibitors against tumor types highly dependent on cholesterol metabolism. SIGNIFICANCE: The presented data indicate, to our knowledge, for the first time, the potential benefit of concomitant modulation of cholesterol biosynthesis pathway and PARP inhibition and highlight the need for further investigation to assess its translational relevance.

Diet suppresses glioblastoma initiation in mice by maintaining quiescence of mutation-bearing neural stem cells.

Glioblastoma is thought to originate from neural stem cells (NSCs) of the subventricular zone that acquire genetic alterations. In the adult brain, NSCs are largely quiescent, suggesting that deregulation of quiescence maintenance may be a prerequisite for tumor initiation. Although inactivation of the tumor suppressor p53 is a frequent event in gliomagenesis, whether or how it affects quiescent NSCs (qNSCs) remains unclear. Here, we show that p53 maintains quiescence by inducing fatty-acid oxidation (FAO) and that acute p53 deletion in qNSCs results in their premature activation to a proliferative state. Mechanistically, this occurs through direct transcriptional induction of PPARGC1a, which in turn activates PPARα to upregulate FAO genes. Dietary supplementation with fish oil containing omega-3 fatty acids, natural PPARα ligands, fully restores quiescence of p53-deficient NSCs and delays tumor initiation in a glioblastoma mouse model. Thus, diet can silence glioblastoma driver mutations, with important implications for cancer prevention.

Growth-rate-dependent and nutrient-specific gene expression resource allocation in fission yeast.

Cellular resources are limited and their relative allocation to gene expression programmes determines physiological states and global properties such as the growth rate. Here, we determined the importance of the growth rate in explaining relative changes in protein and mRNA levels in the simple eukaryote Schizosaccharomyces pombe grown on non-limiting nitrogen sources. Although expression of half of fission yeast genes was significantly correlated with the growth rate, this came alongside wide-spread nutrient-specific regulation. Proteome and transcriptome often showed coordinated regulation but with notable exceptions, such as metabolic enzymes. Genes positively correlated with growth rate participated in every level of protein production apart from RNA polymerase II-dependent transcription. Negatively correlated genes belonged mainly to the environmental stress response programme. Critically, metabolic enzymes, which represent ∼55-70% of the proteome by mass, showed mostly condition-specific regulation. In summary, we provide a rich account of resource allocation to gene expression in a simple eukaryote, advancing our basic understanding of the interplay between growth-rate-dependent and nutrient-specific gene expression.

Differences in Reproductive Success in Young and Old Females of a Long-Lived Species.

Long-lived species are particularly interesting for investigation of trade-offs that shape reproductive allocation and the effective contribution to the next generations. Life history theory predicts that these species will buffer environmental stochasticity via changes in the reproductive investment, while maintaining high adult survival rates. The spur-thighed tortoise was selected as a case study in order to investigate the relationship between the linked maternal characteristics (size and age) and related traits in their hatchlings. We tracked naturally emerging hatchlings from young and old females under semi-natural conditions to test variations in hatchling numbers, body mass, size and survival over two years. We used linear mixed-effect models to analyze variations in hatchling body mass and size, and a mark-release-recapture framework to model their survival. Our study illustrates that old females of long-lived species have greater offspring numbers, greater survival and smaller size when compared with those of young females. The interannual variability evidenced the reduced offspring number and survival in the lower autumn rainfall and spring mean temperature year. Our results highlight the role of maternal age and climatic conditions in the population dynamics and the need for long-term studies of reproduction traits for designating adequate conservation strategies.

Predation of young tortoises by ravens: the effect of habitat structure on tortoise detectability and abundance.

The predation of young tortoise is considered a major cause of mortality for many tortoise species. The predation by common ravens has been identified as being responsible for significant decreases in tortoise populations. Mediterranean spur-thighed tortoise hatchlings and juveniles in Maamora forest (Morocco) were studied in order to describe the size/age class predation of common ravens on young tortoises and infer the drivers of predation risk and population abundance. The results showed a high level of predation on young tortoises (<75 mm carapace length) attributed to ravens in areas with low vegetation cover, representing 100% of the cases of mortality (n = 147), but it was moderate in covered areas (n = 19), representing 12-27%. The population structure of living juveniles differed significantly between covered and uncovered areas, thus suggesting that raven predation might modify juvenile population structure. Finally, N-mixture models showed a positive relationship between (i) bare cover and tortoise detectability that is only evidenced when the plot is far from a perch and (ii) population abundance and shrub species-richness, being higher in uncovered areas. Our results improve the knowledge on predation and survival on this critical stage in life, which is crucial for the conservation of the Mediterranean spur-thighed tortoise.

Implications for Conservation of Collection of Mediterranean Spur-Thighed Tortoise as Pets in Morocco: Residents' Perceptions, Habits, and Knowledge.

The trading and collection of wild animals as pets may be cause for concern regarding animal welfare and species conservation. These concerns can be exemplified by Mediterranean spur-thighed tortoise (Testudo graeca), a long-living species whose use as pets is long established. The human dimension plays a major role in the wildlife for the pet collection, and is particularly important in countries like Morocco, where this might pose a threat to the conservation of the species involved. This study, which is based on a questionnaire survey (n = 480 participants), documents the fact that many people in Morocco keep tortoises as pets: 55% of the participants in the survey and over two tortoises/person. Importantly, most captive tortoises, particularly juveniles, had been collected directly from wild populations by their owners (42%, n = 264). In general, the tortoise owners had limited knowledge of their tortoises' habits and requirements, although rural people were more likely to acknowledge that the tortoise is a wild and threatened species. Our study reveals that non-commercial collection is a common activity in Morocco that may threaten wild tortoise populations and hence species conservation, and it could have consequences regarding the welfare of the animals. We were also able to identify the profile of people towards whom education campaigns should be directed in order to reduce the number of tortoises collected from wild populations. Additional field research should also be conducted to quantify the impact of pet collection on wild tortoise populations.

Size-Dependent Increase in RNA Polymerase II Initiation Rates Mediates Gene Expression Scaling with Cell Size.

Cell size varies during the cell cycle and in response to external stimuli. This requires the tight coordination, or "scaling," of mRNA and protein quantities with the cell volume in order to maintain biomolecule concentrations and cell density. Evidence in cell populations and single cells indicates that scaling relies on the coordination of mRNA transcription rates with cell size. Here, we use a combination of single-molecule fluorescence in situ hybridization (smFISH), time-lapse microscopy, and mathematical modeling in single fission yeast cells to uncover the precise molecular mechanisms that control transcription rates scaling with cell size. Linear scaling of mRNA quantities is apparent in single fission yeast cells during a normal cell cycle. Transcription of both constitutive and periodic genes is a Poisson process with transcription rates scaling with cell size and without evidence for transcriptional off states. Modeling and experimental data indicate that scaling relies on the coordination of RNA polymerase II (RNAPII) transcription initiation rates with cell size and that RNAPII is a limiting factor. We show using real-time quantitative imaging that size increase is accompanied by a rapid concentration-independent recruitment of RNAPII onto chromatin. Finally, we find that, in multinucleated cells, scaling is set at the level of single nuclei and not the entire cell, making the nucleus a determinant of scaling. Integrating our observations in a mechanistic model of RNAPII-mediated transcription, we propose that scaling of gene expression with cell size is the consequence of competition between genes for limiting RNAPII.

Tick parasitism in the Mediterranean spur-thighed tortoise in the Maamora forest, Morocco.

Macroparasites in general, and ectoparasites in particular, have the potential to regulate host population dynamics. In this context, this study addresses the tick parasitism traits of the Mediterranean spur-thighed tortoise (Testudo graeca) in the core area of its distribution range (northwestern Morocco, Maamora). It was discovered that 92.5% of the tortoises were parasitized by ticks in spring, with an infestation intensity and an abundance of 6.7 and 6.2 tick/tortoise, respectively. The observed parasitization rates were among the highest reported worldwide for T. graeca, which could relate to density-depended effects driving host-parasite interactions. The main tick species that parasitized the tortoises were Hyalomma aegyptium (95.6% of the ticks and in the 100% of the parasitized tortoises), Hy. marginatum, Hy. excavatum and Hy. scupense. Individual predictors for the tortoises, such as age, sex and the interaction between body condition and sex, were significantly related to tick abundance. Age-related behavioural differences might favour a higher host-tick effective contact in adults than in juveniles. The fact that males are more active in spring - the breeding season - might explain the observed male-bias in tick abundance and may also be responsible for the negative effect of male body condition on tick infestation rate in contrast to females. Given the potential role played by parasites as regards modulating population dynamics, our results suggest that ticks should be taken into account in the conservation and management programmes of this tortoise species.