Phaidra - University of Veterinary Medicine Vienna
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The impact of cage dividers on mouse aggression, dominance and hormone levels
No full textHome cage aggression in group-housed male mice is a major welfare concern and may compromise animal research. Conventional cages prevent flight or retreat from sight, increasing the risk that agonistic encounters will result in injury. Moreover, depending on social rank, mice vary in their phenotype, and these effects seem highly variable and dependent on the social context. Interventions that reduce aggression, therefore, may reduce not only injuries and stress, but also variability between cage mates. Here we housed male mice (Balb/c and SWISS, group sizes of three and five) with or without partial cage dividers for two months. Mice were inspected for wounding weekly and home cages were recorded during housing and after 6h isolation housing, to assess aggression and assign individual social ranks. Fecal boli and fur were collected to quantify steroid levels. We found no evidence that the provision of cage dividers improves the welfare of group housed male mice; The prevalence of injuries and steroid levels was similar between the two housing conditions and aggression was reduced only in Balb/c strain. However, mice housed with cage dividers developed less despotic hierarchies and had more stable social ranks. We also found a relationship between hormone levels and social rank depending on housing type. Therefore, addition of cage dividers may play a role in stabilizing social ranks and modulating the activation of hypothalamic-pituitary-adrenal (HPA) and hypothalamic-pituitary-gonadal (HPG) axes, thus reducing phenotypic variability between mice of different ranks
Comparison of two peroxidases with high potential for biotechnology applications - HRP vs. APEX2
No full textPeroxidases are essential elements in many biotechnological applications. An especially interesting concept involves split enzymes, where the enzyme is separated into two smaller and inactive proteins that can dimerize into a fully active enzyme. Such split forms were developed for the horseradish peroxidase (HRP) and ascorbate peroxidase (APX) already. Both peroxidases have a high potential for biotechnology applications. In the present study, we performed biophysical comparisons of these two peroxidases and their split analogues. The active site availability is similar for all four structures. The split enzymes are comparable in stability with their native analogues, meaning that they can be used for further biotechnology applications. Also, the tertiary structures of the two peroxidases are similar. However, differences that might help in choosing one system over another for biotechnology applications were noticed. The main difference between the two systems is glycosylation which is not present in the case of APX/sAPEX2, while it has a high impact on the HRP/sHRP stability. Further differences are calcium ions and cysteine bridges that are present only in the case of HRP/sHRP. Finally, computational results identified sAPEX2 as the systems with the smallest structural variations during molecular dynamics simulations showing its dominant stability comparing to other simulated proteins. Taken all together, the sAPEX2 system has a high potential for biotechnological applications due to the lack of glycans and cysteines, as well as due to high stability
A TriAdj-Adjuvanted Chlamydia trachomatis CPAF Protein Vaccine Is Highly Immunogenic in Pigs
No full textChlamydia trachomatis (Ct) infections are the most common sexually transmitted infection (STI). Despite effective antibiotics for Ct, undetected infections or delayed treatment can lead to infertility, ectopic pregnancies, and chronic pelvic pain. Besides humans, chlamydia poses similar health challenges in animals such as C. suis (Cs) in pigs. Based on the similarities between humans and pigs, as well as their chlamydia species, we use pigs as a large biomedical animal model for chlamydia research. In this study, we used the pig model to develop a vaccine candidate against Ct. The vaccine candidate consists of TriAdj-adjuvanted chlamydial-protease-like activity factor (CPAF) protein. We tested two weekly administration options-twice intranasal (IN) followed by twice intramuscular (IM) and twice IM followed by twice IN. We assessed the humoral immune response in both serum using CPAF-specific IgG (including antibody avidity determination) and also in cervical and rectal swabs using CPAF-specific IgG and IgA ELISAs. The systemic T-cell response was analyzed following in vitro CPAF restimulation via IFN-? and IL-17 ELISpots, as well as intracellular cytokine staining flow cytometry. Our data demonstrate that while the IN/IM vaccination mainly led to non-significant systemic immune responses, the vaccine candidate is highly immunogenic if administered IM/IN. This vaccination strategy induced high serum anti-CPAF IgG levels with strong avidity, as well as high IgA and IgG levels in vaginal and rectal swabs and in uterine horn flushes. In addition, this vaccination strategy prompted a pronounced cellular immune response. Besides inducing IL-17 production, the vaccine candidate induced a strong IFN-? response with CD4 T cells. In IM/IN-vaccinated pigs, these cells also significantly downregulated their CCR7 expression, a sign of differentiation into peripheral-tissue-homing effector/memory cells. Conclusively, this study demonstrates the strong immunogenicity of the IM/IN-administered TriAdj-adjuvanted Ct CPAF vaccine candidate. Future studies will test the vaccine efficacy of this promising Ct vaccine candidate. In addition, this project demonstrates the suitability of the Cs pre-exposed outbred pig model for Ct vaccine development. Thereby, we aim to open the bottleneck of large animal models to facilitate the progression of Ct vaccine candidates into clinical trials
Antimicrobial Ionic Liquids: Ante-Mortem Mechanisms of Pathogenic EPEC and MRSA Examined by FTIR Spectroscopy
No full textIonic liquids (ILs) have gained considerable attention due to their versatile and designable properties. ILs show great potential as antibacterial agents, but understanding the mechanism of attack on bacterial cells is essential to ensure the optimal design of IL-based biocides. The final aim is to achieve maximum efficacy while minimising toxicity and preventing resistance development in target organisms. In this study, we examined a dose-response analysis of ILs\u27 antimicrobial activity against two pathogenic bacteria with different Gram types in terms of molecular responses on a cellular level using Fourier-transform infrared (FTIR) spectroscopy. In total, 18 ILs with different antimicrobial active motifs were evaluated on the Gram-negative enteropathogenic Escherichia coli (EPEC) and Gram-positive methicillin-resistant Staphylococcus aureus (MRSA). The results showed that most ILs impact bacterial proteins with increasing concentration but have a minimal effect on cellular membranes. Dose-response spectral analysis revealed a distinct ante-mortem response against certain ILs for MRSA but not for EPEC. We found that at sub-lethal concentrations, MRSA actively changed their membrane composition to counteract the damaging effect induced by the ILs. This suggests a new adaptive mechanism of Gram-positive bacteria against ILs and demonstrates the need for a better understanding before using such substances as novel antimicrobials
Inference of genomic landscapes using ordered Hidden Markov Models with emission densities (oHMMed)
No full textGenomes are inherently inhomogeneous, with features such as base composition, recombination, gene density, and gene expression varying along chromosomes. Evolutionary, biological, and biomedical analyses aim to quantify this variation, account for it during inference procedures, and ultimately determine the causal processes behind it. Since sequential observations along chromosomes are not independent, it is unsurprising that autocorrelation patterns have been observed e.g., in human base composition. In this article, we develop a class of Hidden Markov Models (HMMs) called oHMMed (ordered HMM with emission densities, the corresponding R package of the same name is available on CRAN): They identify the number of comparably homogeneous regions within autocorrelated observed sequences. These are modelled as discrete hidden states; the observed data points are realisations of continuous probability distributions with state-specific means that enable ordering of these distributions. The observed sequence is labelled according to the hidden states, permitting only neighbouring states that are also neighbours within the ordering of their associated distributions. The parameters that characterise these state-specific distributions are inferred.We apply our oHMMed algorithms to the proportion of G and C bases (modelled as a mixture of normal distributions) and the number of genes (modelled as a mixture of poisson-gamma distributions) in windows along the human, mouse, and fruit fly genomes. This results in a partitioning of the genomes into regions by statistically distinguishable averages of these features, and in a characterisation of their continuous patterns of variation. In regard to the genomic G and C proportion, this latter result distinguishes oHMMed from segmentation algorithms based in isochore or compositional domain theory. We further use oHMMed to conduct a detailed analysis of variation of chromatin accessibility (ATAC-seq) and epigenetic markers H3K27ac and H3K27me3 (modelled as a mixture of poisson-gamma distributions) along the human chromosome 1 and their correlations.Our algorithms provide a biologically assumption free approach to characterising genomic landscapes shaped by continuous, autocorrelated patterns of variation. Despite this, the resulting genome segmentation enables extraction of compositionally distinct regions for further downstream analyses
Video Ethogram of Equine Social Behaviour
No full textEquine social behaviour studies face challenges stemming from the absence of a comprehensive ethogram with unequivocal standardised definitions and the resulting limits to data comparison across studies. To address these constraints, this ethogram offers researchers a standardised framework, defining thirty-seven distinct equine social behaviours supplemented by video examples for enhanced clarity. These definitions amalgamate insights from existing ethograms and are fine-tuned through meticulous video observations, encompassing contextual cues such as distinguishing between aggressive and playful circling based on ear position and facial expressions and communicative nuances to provide a detailed representation of equine social behaviours. Video recordings complement the standardised definitions by capturing the dynamic flow and sequence of social interactions. By providing a dynamic and detailed representation, videos allow researchers to observe the temporal aspects of behaviour, including the sequence, duration, and rhythm of interactions. These detailed data are crucial for interpreting social behaviours and unravelling the complexities of equine societies. Standardized and video-illustrated definitions of equine social behaviour facilitate clear and consistent communication between researchers, enabling cross-study comparisons regarding the impact of husbandry practices and health conditions on equine social behaviour, which, in turn, can facilitate the assessment and optimisation of management practices and equine welfare
Redox Cycling of Tetrahedral Iron Drives the Fenton Reactivity of Chrysotile Asbestos
No full textChrysotile asbestos is a carcinogenic fibrous mineral. Its pathogenicity is partly governed by the ability of Fe on the fiber surface to catalyze the Fenton reduction of H2O2 (which is produced during inflammatory processes) to form the highly toxic hydroxyl radical (HO center dot). Recently, tetrahedrally coordinated Fe (Fe-tet) in the fibers\u27 Si sheets was identified as the principal Fe species to catalyze this process. However, as only ferric Fe-tet (Fe-tet(3+)) substitutes Si tetrahedra in chrysotile, Fe-tet needs to redox cycle to ferrous Fe-tet (Fe-tet(2+)) to facilitate fiber-mediated reductions of H2O2 to HO center dot. This redox cycling has never been experimentally investigated. Here we demonstrate, by consecutive ascorbate and O-2 treatments, that structural Fe-tet in exposed Si sheets of chrysotile fibers can redox cycle between Fe-tet(3+) and Fe-tet(2+). Reduction, back-oxidation, and rereduction of Fe-tet(3+) did not labilize the exposed Si sheet and, consequently, did not promote fiber dissolution. However, in the presence of H2O2, prolonged redox cycling of Fe-tet increased fiber dissolution, presumably by accelerating Fe-tet dissolution and subsequent labilization of the exposed Si sheet. Chrysotile fibers for which the concentration of Fe-tet surface sites undergoing redox cycling was lowered through selective Fe removal showed a rebound in Fe-tet(3+) surface site concentration and associated Fenton reactivity once Fe-tet(3+)-depleted Si sheets were dissolved off from the fiber surfaces. To conclude, our results demonstrate that redox cycling of Fe-tet on chrysotile surfaces produces Fe-tet(2+) surface sites, which, as the ultimate Fenton reactive iron species on chrysotile, contribute to the fibers\u27 adverse chemical reactivity
Dogs understand the role of a human partner in a cooperative task
No full textHumans are exceptionally flexible in cooperation, partly due to our ability to recognize the roles of cooperative partners. While some non-human animals understand the need for a partner in such interactions, it is unclear whether they grasp the consequences of their partner\u27s actions and adjust accordingly. Previous studies utilizing economic games with non-human animals yielded mixed results. We investigated dogs, known for their close cooperation with humans, in a stag hunt game. Dogs could cooperate for better rewards or defect for lower ones, while their human partners would either cooperate, never cooperate, or act randomly. We control for attraction to food, side bias, and local enhancement. Dogs were more likely to coordinate with their partners when it led to better rewards, suggesting that they understood their partner\u27s actions. By highlighting this cognitive skill in dogs, we advance our knowledge of the intricate mechanisms driving cooperative behavior in non-human animals
Effect of Salts on Laccase-Catalyzed Polymerization of Lignosulfonate
No full textEnzymatic polymerization of lignosulfonate (LS) has a high potential for various applications ranging from coatings to adhesives. Here, the effect of different ions in low concentrations on enzymatic polymerization of LS was investigated, including salt solutions consisting of mono- and dicarboxylic acids, sulfate, phosphate and chloride with sodium as counter ion. LS polymerization was followed by viscometry and size exclusion (SEC) chromatography. Interestingly, there was only a small effect of ions on the activity of the laccase on standard substrate ABTS, while the effect on polymerization of LS was substantially different. The presence of acetate led to a 39 % higher degree of polymerization (DP) for LS. Small angle X-ray scattering (SAXS) revealed that the structure of the enzyme was largely unaffected by the ions, while the determination of the zeta potential showed that those ions conveying higher negative surface charges onto LS particles showed lower DPs, than those not affecting the surface charge. Further, electron paramagnetic resonance (EPR) spectroscopy showed 5-times higher intensity in phenoxyl radicals for the monovalent ions compared to the divalent ones. It was concluded that the DPs of LS could be tuned in the presence of certain ions, by facilitating the interaction between the laccase substrate-binding site and the LS molecules
Odor cues rather than personality affect tadpole deposition in a neotropical poison frog
No full textAnimals constantly need to evaluate available external and internal information to make appropriate decisions. Identifying, assessing, and acting on relevant cues in contexts such as mate choice, intra-sexual competition, and parental care is particularly important for optimizing individual reproductive success. Several factors can influence decision-making, such as external environmental cues and the animal\u27s own internal state, yet, we have limited knowledge on how animals integrate available information. Here, we used an entire island population (57 males, 53 females, and 1,109 tadpoles) of the neotropical brilliant-thighed poison frog Allobates femoralis to investigate how 2 factors (olfactory cues and personality traits) influence the ability of males to find and use new resources for tadpole deposition. We experimentally manipulated the location of tadpole deposition sites and their associated olfactory cues, and repeatedly measured exploration and boldness in adult males. We further reconstructed tadpole deposition choices via inferred parent-offspring relationships of adult frogs and tadpoles deposited in our experimental pools using molecular parentage analysis. We found that the discovery and use of new rearing sites were heavily influenced by olfactory cues; however, we did not find an effect of the measured behavioral traits on resource discovery and use. We conclude that in highly dynamic environments such as tropical rainforests, reliable external cues likely take priority over personality traits, helping individuals to discover and make use of reproductive resources